Retroviridae

This is a family of large single-stranded RNA viruses that use the enzyme reverse
transcriptase in their replication cycle. A number of oncogenic retroviruses cause animal
diseases of major importance. Because of their mode of replication, retroviruses are
probably present in all vertebrate genomes.
Viral Characteristics
 These RNA viruses are enveloped, single-stranded RNA with an icosahedral
nucleocapsid.
 The envelope has glycoprotein surface spikes. Retroviruses are unique among
viruses in that they bring two identical copies of their genome in virions (are
diploid).
 The ssRNA is converted to ssDNA by the enzyme reverse transcriptase. From the
ssDNA, dsDNA (called provirus DNA) is made, which is then integrated into the
host chromosome. The provirus dsDNA then serves as a template for the
production of mRNA and progeny ssDNA genomes.
 The conversion of ssRNA to ssDNA, mediated by the viral enzyme reverse
transcriptase, results in a dsDNA molecule longer than that of the original
genome. This dsDNA migrates to the nucleus where it is ultimately integrated
into the host chromosome by the viral enzyme integrase.
 Once integrated into the host genome, the viral dsDNA is referred to as a
provirus. The provirus remains latent until "triggered" into transcription of mRNA
by host cell machinery.
 Viral mRNA transcription of the provirus is mediated by cellular RNA
polymerase II.
 The new virions are released by budding, which does not always result in cell
lysis.
 There is a high mutation rate, as reverse transcription is an error-prone process.
Thus, retroviruses usually present a high genetic diversity.
 Many retroviruses carry oncogenes (e.g., Rous sarcoma virus in chickens), while
others do not (e.g., human T-cell lymphotrophic virus). However, some
retroviruses may cause tumors without carrying oncogenes.
 All retroviral genomes consist of two molecules of ssRNA, (+)sense, have 5' cap
and 3' poly-(A) (equivalent to mRNA) and four characteristic coding regions
(gag-pro-pol-env). Gag (group specific antigen: matrix protein, nucleoprotein,
capsid) genes; pro (protease) gene; pol (reverse transcriptase and RNase-H); and
env (envelope, receptor binding) genes. These vary in size from ~8-11 kb. They
are the only viruses that are truly diploid. Additionally, there is a specific type of
cellular transporter RNA (tRNA) (usually trp, pro or lys) - required for replication
that is present in the virion.
 The virions are sensitive to heat, lipid solvents, and detergents but are relatively
resistant to ultraviolet light damage.

Classification
Currently there are seven genera in the family. All of the genera except Lentivirus and
Spumavirus cause neoplastic changes in specific cell types. The genera are, with
significant viruses, as follows:
  Alpharetrovirus
o Avian leukosis virus
o Avian sarcoma virus
o Avian myeloblastosis virus
o Rous sarcoma virus
 Betaretrovirus
o Mouse mammary tumor virus: A much studied important virus of mice.
o Ovine pulmonary adenocarcinoma virus
o Enzootic nasal tumor virus
 Gammaretrovirus
o Feline leukemia virus
o Feline sarcoma virus
o Avian reticuloendotheliosis virus
 Deltaretrovirus
o Bovine leukemia virus
 Epsilonretrovirus
o Fish tumor viruses
 Lentivirus These viruses cause immunodeficiency-type diseases.
o Maedi/Visna virus
o Caprine arthritis encephalitis virus
o Equine infectious anemia virus
o Feline immunodeficiency virus
o Bovine immunodeficiency virus
o Human immunodeficiency viruses 1 and 2
o Simian immunodeficiency viruses
 Spumavirus Foamy agents (e.g., bovine and simian foamy viruses) that cause
persistent but silent infections in several natural hosts. They are occasional
contaminants in cell cultures.
The major diseases caused by retroviruses are discussed below under their genera.

Alpharetrovirus
Avian Leukosis
Cause
Avian leukosis virus. The avian leukosis group of viruses (ALGV) is comprised
of 10 subgroups, A through J, based on differences in envelope glycoproteins. Subgroups
A to E, and J viruses infect chickens. The other subgroup viruses occur in quail,
partridges and pheasants.
The ALGV group contains both replication-competent and replication-defective
viruses. The defective viruses are not important as causes of leukosis.
Subgroup A viruses cause most outbreaks of avian leukosis that manifest as a lymphoid
leukosis, a B cell lymphoma. This is the most important and common neoplasm
associated with the disease.
Viruses of subgroup J have been associated with myeloid leukosis in broilers.
Most of the viruses causing avian leukosis are exogenous. Endogenous ALGVs, which
mostly belong to subgroup E and occur commonly in chickens and other avian species,
are of little or no pathogenicity. These proviral DNA sequences are integrated into germ
line cells and are thus transmitted vertically.
Occurrence
Avian leukosis viruses occur commonly in chickens, quail, partridges and pheasants
worldwide and the disease is of great economic importance.
Transmission
Although transmission may occur by contact, as virus is present in feces and
saliva, the principal means of transmission is via the egg. Blood-sucking parasites are
potential vectors. Endogenous viruses, which are common in the chicken but not
important as causes of leukosis, are also transmitted genetically in the germ line from
parent to offspring.
Pathogenesis, Clinical and Pathologic Features
All avian leukemia group viruses are oncogenic, except those belonging to
subgroup E. The mechanisms involved in the development of neoplasia by viruses, that
is, oncogenesis. Such factors as genetics (autosomally transmitted susceptibility or
resistant avian cells) and the age and sex of the host can also be important in the
development of tumors.
The incubation period is variable and may be as long as months or years. After
infection the virus is disseminated to tissues throughout the body where it replicates.
Within B-cells, the provirus induces neoplasia by integrating closely to the host c-myc
oncogene. As a result, the cell is stimulated to divide when the viral genes are being
transcribed. In erythroblastosis, integration is near the c-erbB gene, with a similar
outcome.
With those viruses that are considered fast-transforming, their genome has been
found to contain the c-onc proto-oncogene. Multiple copies of the c-onc gene result in
overproduction of the protein, which stimulates cellular transformation. The oncoproteins
produced may act as hormones, growth factor receptors, transcription regulation factors,
or kinases associated with cellular signaling mechanisms.
Viruses of subgroups A through D are associated with a variety of neoplastic conditions
in chickens, including lymphoid leukosis, erythroblastosis, myeloblastosis,
myleocytoblastosis, osteopetrosis, nephroblastomas, hemangiomas, and sarcomas. The
more common forms are as follows:
 Lymphoid leukosis is by far the most prevalent form of avian leukosis / sarcoma.
The disease is most often seen in chickens at least 16 weeks of age and occurs
more commonly in females. Clinical signs may be absent or birds may appear
thrifty with pale combs. The abdomen may be enlarged be cause of massive tumor
growth in the liver.
 Osteopetrosis is a sporadic disease that occurs primarily in males. The shafts of
the long bones are thickened, often resulting in a stilted gait. Affected birds may
be anemic and also have lesions of lymphoid leukosis.
 Erythroblastosis may be manifested in one of two forms, anemic or proliferative,
with the latter form being the more common. Clinical signs are similar for both
forms. Chickens become depressed, emaciated, and dehydrated. With the anemic
form, there are few circulating erythroblasts and chickens appear pale, as opposed
to cyanotic with the proliferative form in which circulating erythroblasts are
present in large numbers.
  Myeloblastosis is similar clinically to erythroblastosis. There is an abnormal
proliferation of myeloblasts resulting in severe leukemia.
Diagnosis
 Clinical specimens: whole birds in extremis.
 Diagnosis of the neoplastic conditions caused by avian leukosis / sarcoma viruses
is usually based on clinical signs and gross and histopathologic lesions, but
lymphoid leukosis may be confused with the acute form of Marek's disease in
older birds.
 Differential features, nodular tumors in the bursa of Fabricius as opposed to
diffuse enlargement with Marek's disease; intrafollicular cell proliferation in the
bursa of Fabricius as opposed to interfollicular cell proliferation with Marek's
disease; and the cytologic appearance of lymphoid cells that are uniformly "blast"
cells with lymphoid leukosis but a mixture of mature and immature pleomorphic
cells with Marek's disease.
 Isolation of the virus or the demonstration of antibody is not considered to be of
value in diagnosis because avian leukosis group viruses are ubiquitous in
chickens.
Prevention
 There is no vaccine available and eradication is the preferred method of control.
Since the viruses are primarily transmitted via the egg, virus infected breeders are
detected by testing the egg albumin for viral antigen by an ELISA method.
Positive birds are eliminated.
 ELISA and immunofluorescence assays are used to detect antibodies in serum and
egg yolk.
 Most commercial flocks of chickens are now free of exogenous ALG viruses.
 Some birds are genetically more resistant to fowl leukosis; they have decreased
numbers of specific cell surface viral receptors.
 Incubators should be clean and adequately disinfected.
 Control blood-sucking parasites.

Rous Sarcoma Virus and other Alpharetroviruses

The Rous sarcoma virus (RSV), avian myeloblastosis virus and avian erythroblastosis
virus are frequently defective and require a helper ALGV for replication. These viruses
can become rapid transforming viruses (experimentally) when a cellular oncogene is
incorporated into their genome. They are not thought to be transmitted naturally. The
RSV was the first virus, shown in 1911 by Peyton Rous, to cause a solid malignant
tumor. Subsequently it was found that there were a number of RSVs that produce tumors
in various mammals.
The defective avian sarcoma virus carries the transduced oncogene v-crk, whose
product binds to several cellular proteins resulting in in vitro cell transformation.

Betaretrovirus
Ovine Pulmonary Adenocarcinoma
(Jaagsiekt, Pulmonary adenomatosis)
Cause
Ovine pulmonary adenocarcinoma virus. As many as 15 - 20 copies of
endogenous virus per cell are present in the genome of sheep; however, disease is caused
by exogenous virus.
An oncogene has not been found in the genome and the mechanism of
transformation has not been elucidated.
Occurrence
The disease occurs in sheep worldwide except for Australia. It rarely affects goats.
Transmission
By airborne respiratory droplets and also by vertical transmission.
Clinical and Pathologic Features
The virus replicates in alveolar and bronchial cells and tumors arise from these
cells. Pulmonary adenomatosis is a chronic slowly progressive, eventually fatal
pneumonia of sheep characterized by adenomatous proliferation of viral infected alveolar
and bronchiolar epithelium. The resulting adenocarcinomas may metastasize.
Mannheimia or Pasteurella bacteria may be involved in a secondary pneumonia, which
may be terminal.
There is coughing, nasal discharge, and difficulty breathing, leading eventually to
emaciation and death. The incubation period may be as long as several years.
The disease can be confused clinically with ovine progressive pneumonia (discussed
below - Maedi/Visna), but lung lesions are distinctly different. Grossly, there are
widespread nodular growths throughout the lung that are microscopically adenomas or
adenocarcinomas.
Diagnosis
 Clinical specimens: Formalin-fixed lung tissue with lesions.
 Diagnosis is usually based on gross and microscopic lung lesions.
 Attempts to cultivate the virus in cell cultures have not been successful.
 ELISA and PCR have been used to detect virus in pulmonary exudates.
 The absence of detectable specific antibodies precludes the use of serological
tests.
Prevention
 Vaccines are not available.
 Clinically affected sheep should be removed from the flock. The disease was
eradicated from Iceland by depopulation measures.

Enzootic Nasal Tumor

The enzootic nasal tumor virus of sheep and goats is closely related to the virus causing
ovine pulmonary adenocarcinoma. The genomes of sheep and goats have copies of
endogenous betavirus sequences that are closely related.
The disease is characterized clinically by serous to mucopurulent nasal discharge and
open-mouthed breathing. The tumor or tumors originate from the mucosa of the nares and
may be unilateral or bilateral. The nasal passages may become completely occluded, and
the tumorous growths may extend into the sinus, cranial cavity, and pharynx.
Diagnosis is based on gross and microscopic lesions. Confirmation requires
demonstration of the virus.
 Gammaretrovirus
Feline Leukemia (FeL)
Cause
Feline leukemia virus (FeLV).
 There are three subgroups of the virus, A, B and C based on differences in the
gp70 envelope glycoprotein.
 Subgroup A viruses grow exclusively in feline cells; B and C viruses also grow in
human, canine and mink cells. The viruses can be propagated for long periods in
feline fibroblast cells; replication does not result in observable CPE.
 FeLV-A can be recovered from all cats naturally infected with viral feline
leukemia. FeLV-A tends to be less pathogenic than the other two subgroups.
 FeLV-B arises as a result of recombination between env genes of FeLV-A and
proviral DNA of endogenous FeLV. FeLV-B can be isolated from about 50% cats
with FeL.
 FeLV-B is only transmitted with FeLV-A. FeLV-B is frequently lost in cats with
FeL.
 Infection with both subgroups A and B viruses is more serious than with only
subgroup A. For example, neoplastic disease incidence increases with A and B
coinfection over that of A alone. Among those chronically infected with subtypes
A and B, 30% are diagnosed with lymphoma.
 FeLV-C arises in FeLV-A infected cats as a result of mutations in the FeLV-A
env gene.
 FeLV-C causes a fatal anemia and thus is rarely transmitted.
 Frequent recombination of FeLV with host cell genes yields replication defective
recombinant feline sarcoma viruses. The latter viruses can be isolated from
fibrosarcomas in young cats.
 FeLV possess the characteristic retrovirus gene arrangement, where LTR stands
for lateral terminal repeats.
 FeLV has been associated with a variety of different malignancies. The
mechanisms of oncogenesis include: possession of viral oncogenes, such as v-
myc; transduction of a proto-oncogene to an active state; disruption of a tumor
suppressor gene; or insertional mutagenesis (insertion of proviral DNA disrupts
the structure or function of another gene, generally involved in cell cycle
regulation).
Occurrence
The disease, which occurs worldwide, is a major, frequent malady of domestic cats and
some other Felidae including wild cats and jungle cats.
Colostrum appears to protect kittens for the first month of life. Older kittens are
particularly susceptible with their susceptibility decreasing with age. It is estimated that
only about 1 in 5 cats exposed at 10 weeks of age will develop persistent infection.
Kittens of persistently infected queens usually become infected.
Transmission
The virus is present in respiratory and oral secretions and in urine and feces. Infection is
by ingestion and spread is by direct and indirect contact and transplacentally. Saliva is
particularly infectious and biting and mutual grooming are common means of spread.
Pathogenesis
The main stages in the progression of FeLV infection are as follows:
 Initial infection Virus replicating in surrounding lymphatic tissues
 Primary or transient viremia Replication in systemic lymphatic tissues, bone
marrow and other tissues (May be elimination of the virus or latent infection.)
 Secondary or persistent viremia (May be elimination of the virus)
Extensive involvement of the bone marrow, stomach, pharynx, esophagus,
salivary glands, bladder, and respiratory tract leading to one of the following:
Virus elimination / Latent infection / Active infection
 With active infection, virus is excreted in saliva, feces, urine, and respiratory
secretions.
 Clinical disease develops and is manifested in various forms.
Clinical and Pathologic Features
Most cats when infected with FeLV mount an effective immune response and recover
completely without evidence of any clinical signs. A small percentage is viremic for a
variable period but are asymptomatic and eventually eliminate the virus.
Those whose immune response is insufficient (about 2%) remain persistently infected
and may go on to develop one of a number of neoplastic or non-neoplastic forms of the
disease.
Approximately 80% of cats harboring the virus die within three years.
Clinical signs vary with the different forms of the disease and are related to the nature,
extent and location of lesions. The forms are as follows:
Neoplastic forms:
Approximately 20% of persistently infected cats develop one of the following
lymphosarcoma forms: alimentary, multicentric, thymic, or lymphoid leukemic. Clinical
signs vary with the different forms of tumors. General signs are lethargy, anorexia and
weight loss.
Some important features of the various forms of lymphosarcoma are as follows:
 Alimentary form: The cat may display anorexia, vomiting and diarrhea.
Abdominal masses involve the small intestine, cecum and colon; associated
mesenteric lymph nodes may be affected.
 Multicentric form: Generalized lymphoadenopathy, renal lymphosarcoma,
splenomegaly, and hepatomegaly may be found. This form is usually seen in
young cats.
 Thymic form: Dysphagia, and dyspnea are common signs, and cyanosis may be
present in advanced cases. Pleural fluid may contain neoplastic cells.
 Lymphoid leukemic form: The bone marrow is primarily involved and
cancerous lymphocytes circulate in the blood. Jaundice, fever anemia and pallor
of mucous membranes are frequent, and lymphadenopathy, splenomegaly, and
hepatomegaly may be present. Varying degrees of fever, anorexia and weakness
are evident.
 Myeloid leukemia: The primary lesion of this non-lymphosarcoma form is in the
bone marrow with secondary involvement in the liver, spleen and lymph nodes.
This form of leukemia is named according to which hematopoietic cell line is
affected, e.g., myelogenous leukemia, erythroleukemia, and lymphoblastic
leukemia. Signs include progressive anemia, recurring fever and weight loss.
It should be kept in mind that not all cats with the above referred to forms of FeLV
infection will be serologically positive for FeLV antigen. This may complicate the
diagnosis.

Non-neoplastic forms:
Immunosuppression
The mechanism responsible for FeLV induced immunosuppression is not well
understood. It is thought that the envelope protein p15E may be involved.
The immunosuppression increases susceptibility to bacterial, fungal, protozoan, and viral
agents. Some of the manifestations are as follows:
 There may be chronic recurring rhinitis and sinusitis, sores around claws, and
peridontal disease. However, these may also be seen as a result of feline
immunodeficiency disease. The healing of infectious processes, including
abscesses, may be delayed in cats with FeLV infection.
 Cats infected with FeLV are particularly susceptible to bacterial, fungal, and viral
respiratory and enteric pathogens. In the chronic infections that ensue there is
persistent fever with a progressive loss of weight and condition.
 FeLV infection may predispose to feline infectious peritonitis and
Haemobartonella felis infection (feline infectious anemia).
 A panleukopenia-like syndrome has been associated with FeLV infection. It has
occurred in cats immunized against feline panleukopenia and is invariably fatal.
Reproductive Disorders
 FeLV infection may result in fetal death (fetal resorption), abortion (late
gestation) and infertility. Fetal death is thought to be due to endometritis and
placentitis. It is estimated that about 75% of infected queens abort.
 Fetuses that survive to term are persistently infected and the resulting kittens are
weak and sickly. FeLV infection is considered a cause of what is called fading
kitten syndrome.
Glomerulonephritis
This may be present in cats with persistent FeLV infection. It is thought to be due to the
deposition of antigen-antibody complexes in the kidney. There is evidence that this
immune complex-mediated glomerular nephritis is an important cause of death in FeLV
infection.
Diagnosis
 Clinical specimens: Blood smears, tumors, bone marrow, and serum.
 A number of diagnostic procedures can be used including histopathologic
examination of biopsies, bone marrow examinations, and cytology of thoracic and
abdominal fluids. However, the most practical and reliable way to diagnose FeLV
infection is with the ELISA and IFA procedures referred to below. All three
subgroups are detected by the commonly used FeL diagnostic tests, but they
cannot distinguish between the various subtypes.
 An indirect fluorescence antibody procedure (IFA) on blood smears and the
ELISA procedure on serum are the procedures most commonly used for the
detection of antigen (the major capsid protein, p27). This antigen can be found in
large amounts in the cytoplasm of infected leukocytes and in platelets. The
soluble form is found in the plasma and serum of infected cats. At least three
 blood smears are recommended for the IFA. A positive test indicates the presence
of virus.
 The results of the IFA test and ELISA compare favorably.
 Commercial reagents are available to diagnostic laboratories for the ELISA and
the IFA tests. Commercial ELISA kits are available for practitioners.
 The virus can be readily propagated in cell cultures, but virus isolation as a
diagnostic tool is expensive and time consuming.
 PCR can be used to detect the virus genome.
Treatment
 Supportive therapy.
 Chemotherapy and irradiation may prolong life in cats with neoplasia.
 Antiviral agents (including human recombinant INF-α and AZT) may delay the
onset of clinical signs but are not curative.
Prevention
 The FeLV is labile and quickly loses its infectivity in the environment. It is easily
inactivated by commonly used disinfectants.
 The disease can be eradicated from colonies by periodic serologic testing with the
removal of positive cats, and disinfection of potentially contaminated areas. New
additions should be quarantined and tested before admission to the main colony.
Cats in the main colony are retested at yearly intervals or less.
 There should be an interval of at least 1 month before introducing negative cats to
a formerly infected environment.
 Some owners will elect to keep FeLV-positive cats that are asymptomatic. Such
cats are a threat to negative cats and thus should be kept indoors and isolated from
negative cats. They may later develop feline leukemia virus-related disease.
 Subunit and killed virus vaccines are available and administered from nine weeks
of age. They do not eliminate pre-existing infections. The vaccines don't interfere
with tests for viral antigen.
 Evaluation with ELISA or IFA is recommended prior to vaccination.

Feline Sarcoma Virus

Feline leukemia viruses often undergo recombination with host cell genes resulting in
replication-defective recombinant sarcoma viruses (FeSV). They are unable to replicate
without the aid of wild-type FeLV. In each of these recombinant viruses one of seven
oncogenes has been transduced by FeLV. For example, the Gardner-Arsstein-GA-FeSV
has the (c-fas) oncogene. More than ten different feline sarcoma viruses have been
identified.
Feline sarcoma viruses have been isolated from rare multicentric fibrosarcomas in
younger cats. FeSV is confined to the tumor and there is no evidence of cat-to-cat
transmission of these viruses.
Definitive diagnosis depends on extensive laboratory procedures.

Avian Reticuloendotheliosis
This term comprises several disease syndromes. The causes are reticuloendotheliosis
(RE) viruses (Gammaretrovirus) mainly comprising three closely related serologic
subtypes that are distinctly different from the leukosis/sarcoma group of retroviruses.
These viruses are widespread and may cause various serious neoplasia in chickens and
turkeys, and also in ducks, geese, pheasants and Japanese quail. Some outbreaks have
been attributed to vaccines contaminated with reticuloendotheliosis virus.
Virus spread occurs through contact and via the egg. Infections are frequently subclinical.
Viral antigen in serum can be detected with an agar gel precipitin test. ELISA
(commercially available) and other serological procedures are used to detect antibody.
The viruses can be cultivated in a variety of avian cells but often replicate without
discernible cytopathic effect.
Given the nature of disease control measures are not feasible.

Deltaretrovirus
Bovine Leukosis
(Bovine leukemia, enzootic bovine leukosis)
Cause
Bovine leukemia virus (BLV). Only one antigenic type has been found. Only a
small percentage of animals infected with BLV develop B cell lymphosarcoma. Most
infected animals develop a persistent lymphocytosis without any apparent clinical
features.
The virus produces syncytia in cell cultures and immune serum prevents syncytia
formation. This is used as an assay for virus or antibody.
Occurrence
The bovine leukemia virus (BLV) is distributed worldwide and occurs particularly
frequently in dairy cattle. Most infections are unapparent and as many as 80% of a dairy
herd may be infected. The disease has been eradicated from some countries; others are in
process of eradication.
Transmission
Cattle are infected by direct contact with blood from an infected animal during
blood sampling, clinical examinations, castration and dehorning. Mechanical
transmission by biting insects may play a role, particularly in tropical regions.
Approximately 10 to 15% of calves born to infected cows are infected.
Pathogenesis
B lymphocytes are the primary target cells and there follows an unapparent
infection or a B-cell lymphoma. The virus does not contain an oncogene. Instead two
regulatory gene proteins, Tax and Rex, are key in the progression of neoplasia. In
particular, the Tax protein associates with host cell transcription factors leading to the
transcription of the BLV provirus.
Most infected animals are asymptomatic; around 30% develop a persistent
lymphocytosis and less than 2% eventually develop lymphosarcoma.
Clinical and Pathologic Features
Although non-immune cattle of all breeds can be infected at any age with bovine
leukemia virus (BLV), most infections occur in dairy cattle more than two years of age.
The fact that disease is seldom seen in younger animals is related to the presence of
protective maternal antibody (for 5 - 6 months) and the separation of younger animals
from the remaining herd until they reach sexual maturity.
Most animals infected with BLV remain clinically normal. Those that develop disease
(approximately 2%) ultimately die after a long (weeks to months) clinical course. Initial
clinical signs are often those of weight loss and reduced milk production, but may be
quite varied depending on the site of tumor development.
The disease is a B-cell lymphoma. The organs most commonly affected are the lymph
nodes, heart, abomasum, uterus, and spleen. When the superficial lymph nodes are
involved they may be swollen and appear as lumps under the skin usually in the neck and
rear flank region.
Diagnosis
 Clinical specimens: Affected tissues (formalin-fixed) and serum.
 Presumptive diagnosis of BL is often based on the finding of tumors in the
locations mentioned above upon clinical and gross necropsy examination.
Microscopic examination of affected tissues is required to confirm the disease. It
must be kept in mind that there are sporadic forms of non-viral bovine leukosis
(thymic, multicentric); they usually affect younger animals.
 Specific antibody may be detected in the serum of infected cattle by different
serological tests including ELISA, radioimmunoassay and the agar gel
immunodiffusion test (AGID). The latter is most commonly used for diagnosis
and epidemiological studies. Many cattle have antibodies to BLV but most never
develop disease.
 The virus can be isolated and cultivated in blood lymphocytes but this is not
practicable for diagnosis.
 Although not practicable for diagnosis PCR can be used to detect virus in
lymphocytes.
 Blood lymphocyte counts, which were once used in diagnosis, are no longer used
as not all infected animals have a lymphocytosis.
Prevention
 Vaccines to prevent BL are not available.
 Eradication is accomplished by testing and removal of serologically positive
animals and only admitting to the herd serologically negative cattle.
 Care must be taken to prevent spread of the virus during blood sampling, clinical
examinations, castration, dehorning, etc. Insect control may be advisable in
affected areas.
 In heavily infected herds eradication may not be feasible. Negative animals
should be separated from the serologically positive and efforts directed to
preventing spread. By taking adequate precautions it is possible to maintain herds
with a few seropositive animals, without spread to other animals.

Lentivirus
Maedi/Visna
(Ovine progressive pneumonia)
Cause
Maedi/visna (MV) virus, a species in the genus Lentivirus ovine/caprine lentivrus
group. The virus has a dense core and the RNA genome is related to that of the virus of
caprine arthritis encephalitis. There are homologies of 60% in the env gene and about
75% in the gag and pol genes. Antigenic changes take place that are thought to be due to
selection of strains expressing alternative antigens.
Occurrence
The disease occurs widely in sheep throughout the world with the exception of
Australia, New Zealand, and Iceland from which it was eradicated.
Transmission
The virus of MV is shed in nasal secretions and in the milk of infected ewes.
Pathogenesis
The incubation period varies from several months to years. Following initial
infection, most animals are viremic. This allows for the development of an immune
response, both humoral and CMI. However, this is insufficient for elimination of the
infection and facilitates the development of characteristic immunopathology.
Infection results in a chronic, progressive inflammation, which is characterized by the
presence of macrophages and lymphoproliferation mainly in the lungs and mammary
glands. Lesions also occur in synovial membranes and the brain. These lesions are the
result of immunopathology in response to the presence of persistent viral antigens.
Activation of latent provirus occurs in monocytes, triggered by monocytes maturation
into macrophages in response to inflammatory signals. Therefore, infected monocytes are
a source of persistent antigen – only expressing viral antigen whenever they mature into
macrophages.
In addition to the persistence associated with latent virus-infected monocytes, the
replication of ssRNA for progeny genomes is error-prone. As a result, many variants of
the virus exist within an individual. The immune system then needs to respond to the
various viral variants, as this takes time the variants become a source of persistent
antigen.
Clinical and Pathologic Features
Maedi/Visna is a slowly progressive, eventually fatal disease in a small
percentage of adult sheep. It is manifested in two different forms, the respiratory
syndrome (with arthritis and mastitis) and the infrequent nervous syndrome. The
Icelandic words "maedi" and "visna" mean dyspnea and wasting, respectively.
Although sheep remain infected for life, few develop clinical disease. In those that do,
there is a protracted incubation period of a year or longer. The various disease
manifestations, which progress over a period of months, are most often seen in sheep
more than two years of age.
The pulmonary form is characterized by a slow and progressive weight loss and
dyspnea upon exertion. Sheep with the CNS demyelinating form develop ataxia of the
hind quarters and ultimately paralysis.
Chronic active inflammation is noted histologically in affected tissues. Lung
lesions are those of interstitial pneumonitis and lymphoid hyperplasia.
Leukoencephalomyelitis with focal demyelination and lymphocytic perivascular cuffing
are observed in the brain. Affected mammary glands are fibrotic with lymphocytic
infiltrates, and affected joints are characterized by proliferative changes in the synovial
membrane.
Diagnosis
 Clinical specimens: Serum and live clinically ill sheep.
 Diagnosis is usually based on clinical signs and histopathologic lesions with
supporting serologic evidence of exposure as determined by the agar gel
immunodiffusion (AGID) test, ELISA or Western blotting.
  The virus of MV is closely related antigenically to the virus of caprine arthritis
encephalitis and the current AGID test kits will detect antibodies to both viruses.
 Virus isolation is not practicable for routine diagnosis.
Prevention
 No vaccines are currently available.
 Prevention is best accomplished by maintaining closed serologically negative
flocks. All replacement stock should be serologically negative.
 Control of MV within a flock requires periodic serologic testing and subsequent
removal of seropositive animals. Lambs should not be allowed to suckle
seropositive ewes.

Caprine Arthritis Encephalitis

(Leukoencephalomyelitis of goats, caprine encephalomyelitis)
Cause
Caprine arthritis encephalitis virus; a non-oncogenic retrovirus of the genus
Lentivirus. As mentioned earlier, this virus is closely related antigenically to the virus
causing Maedi/Visna.
Occurrence
Caprine arthritis-encephalitis (CAE) virus is widely distributed in goats of all ages
and breeds throughout the world. The disease is seen most frequently in dairy goats.
Transmission
Young kids are most commonly infected through the ingestion of milk from
infected does. Horizontal transmission is thought to occur by direct contact with
secretions and excretions from infected goats and through coitus.
Pathogenesis
The pathogenesis of the disease is similar to Maedi/Visna. It is characterized by
chronic persistent inflammation, which is not resolved in spite of a vigorous CMI
response to viral antigen. Infected monocytes are a source of persistent antigen and a
large number of viral variants generated during viral replication; this combination
contributes to the inability of the host to resolve the infection. Immunopathologic
mechanisms are thought to be responsible for the development of most lesions.
Clinical and Pathologic Features
The development of the disease is similar to Maedi/Visna. Virus replicates in
macrophages and CMI responses contribute to the lesions. Chronic connective tissue
disease affects the joints of mature goats; a leukoencephalomyelitis is seen in young
goats and less commonly there is mastitis or pneumonia in older goats.
The arthritic form is characterized by a proliferative synovitis of the carpal, fetlock, hock,
and stifle joints. Joints are enlarged, especially the carpal joints, and common histological
lesions are those of synovial cell proliferation and inflammatory cell infiltration
(lymphocytes, plasma cells, and macrophages). The arthritis may remain static or become
progressively worse with development of fibrinous concretion, necrosis, and
mineralization.
Central nervous system (CNS) disease is principally seen in young kids 2 - 4
months of age and is characterized by a progressive ascending paralysis. Microscopically,
the CNS lesions resemble those of Visna in sheep and include lymphocytic cell
infiltration and demyelination of the white matter.
Some infected goats may develop an interstitial pneumonitis and some infected does may
develop swollen, fibrotic mammary glands.
Diagnosis
 Clinical specimens: Serum and affected sheep.
 Diagnosis of CAE associated disease is usually made on the basis of clinical signs
and serologic evidence of exposure. The serologic tests most often used are the
agar gel immunodiffusion procedure and ELISA.
 The virus can be propagated in caprine cell cultures derived from synovial
membranes, but isolation is time consuming and rarely attempted. The virus
induces syncytia in cultured cells although it may take weeks for them to become
obserable.
Prevention
 Goats infected with CAB virus remain infected for life.
 Prevention is best accomplished by maintaining closed herds.
 All replacement animals should be isolated and tested serologically (ELISA or
AGID).
 Control of CAE in herds in which the virus is enzootic is difficult. If
economically feasible, test and removal is the preferred method. Alternatively,
serologically positive animals should be segregated from negative animals. Young
kids should be removed from positive does at birth.
 Milk from infected does should not be fed unless it has been pasteurized.

Equine Infectious Anemia

(Swamp fever)
Cause
Equine infectious anemia virus (EIAV). Strains have a common CF antigen but can be
differentiated by virus neutralization. EIAV is capable of infecting all members of
equidae, including horses, mules and donkeys. It was also the first retrovirus observed to
be transmitted mechanically by insects (tabanids). Infected animals remain infected for
life.
Occurrence
Equine infectious anemia (EIA) is a disease of Equidae that occurs frequently in many
countries. It is particularly prevalent in low-lying swampy regions. Incidence of the
disease has been greatly reduced in some countries as a result of control measures.
Transmission
This occurs mechanically when blood from an infected horse is introduced into a
susceptible horse via hematophagous insects (tabanids), hypodermic needles, and surgical
instruments. The virus may be present in milk, saliva, semen and urine and thus direct
and indirect transmission is thought to be possible.
Pathogenesis
EIA virus infects monocytes/macrophages and Kupffer cells, whereby the integrated
provirus can be transported throughout the body. In spite of a vigorous immune response
to viral antigen, the virus persists as provirus within infected cells.
As with other retroviruses, many variants of the virus are produced due to the error-prone
replication process. In the case of EIA, release of these variants can be traced to
intermittent febrile periods.
 The humoral response to the presence of the viral antigen stimulates the
production of non-neutralizing antibodies. These antibodies bind viral antigen, resulting
in the formation of immune complexes. Once formed, the immune complexes trigger the
classical complement cascade, which in turn leads to fever production, anemia,
thrombocytopenia, and glomerulonephritis.
The anemia is the result of the combined effects of hemolysis, phagocytosis of
erythrocytes, and a decrease in the production of the blood cells of the erythropoietic
lineage.
Clinical and Pathologic Features
The incubation period following exposure to EIA virus is generally 2 - 6 weeks. The
resulting disease may be acute or chronic.
The acute form is characterized by sudden onset of fever, depression, anorexia,
thirst, progressive weakness, petechial hemorrhages, ventral edema, and death in 2 - 4
weeks.
The chronic form of EIA is the most common. Affected horses are intermittently febrile
and anemic. They experience weight loss and often have edema of the limbs and
abdomen. Some affected horses may exhibit ataxia, and on rare occasions ataxia may be
the only clinical sign. Remissions are common and may last for years with the animal
appearing essentially normal. Viremia may be present for years and even during
remissions, however, the disease is usually fatal.
Diagnosis
 Clinical specimens: Serum, cerebrospinal fluid from ataxic horses.
 Since EIA infection is persistent, the demonstration of specific antibody (to the
core virus protein p26) is routinely used for diagnosis. This is accomplished most
reliably by the agar gel immunodiffusion test (AGID) usually referred to as the
Coggins test.
 A competitive ELISA is also used but results should be confirmed with the
Coggins test.
 The virus can be isolated and cultivated in equine leukocyte culture; there is no
noticeable CPE.
 Proviral DNA can be detected by PCR. Additionally, viral RNA can be detected
by reverse transcriptase (RT)-PCR. In RT-PCR, the RNA template is copied into
cDNA by the enzyme reverse transcriptase (isolated from a retroviral source),
using primers specific to the genome of the virus.
Prevention
 Vaccines are not currently available.
 All horses on the premises should be serologically tested. Positive horse should be
destroyed or confined to vector proof quarters, or segregated from other horses at
pasture by at least 200 yards to prevent mechanical spread by biting insects.
 Many countries and states will only allow entrance of sero-negative animals.

Feline Immunodeficiency Virus Infection

Cause
Feline immunodeficiency virus. Five subtypes of FIV have been identified based on
differences in the envelope amino acid sequences.
Occurrence
Feline immunodeficiency virus (FIV) occurs frequently in cats throughout the world.
Similar viruses have been recovered from wild and zoo felids. Male free-roaming cats
have the highest incidence of infection.
Transmission
This is mainly by bites.
Pathogenesis
The pathogenesis of FIV infection is similar to that observed with HIV, and thus FIV
infection is sometimes referred to as feline AIDS. FIV replicates primarily in CD4+
(helper) T lymphocytes, but also in macrophages, astrocytes, and microglial cells.
Once infected, a chronic viremia ensues that peaks at 7 - 8 weeks then declines. However,
viremia increases in the terminal stages of the illness.
The humoral response is normal. However, there is progressive decrease in the function
of the CMI due to decreases in CD4+ cells. As a result of the CD4+ decrease, an increase
in the numbers of opportunistic infections associated with clinical immunodeficiency is
observed.
There is also an increase in the number of variants, due to error-prone replication.
Clinical and Pathologic Features
The disease is seen in three stages, as follows:
 Acute stage: Clinical signs appear 1 - 2 months following infection and consist of
depression, fever, and a generalized lymphadenopathy.
 Subclinical stage: Signs of illness disappear after a few weeks or months but cats
remain viremic for life. A period of relative normalcy may last for several months
to years.
 Chronic stage: Infected cats eventually succumb to various chronic opportunistic
infections be cause of an impaired immune system. Chronic infections may result
in stomatitis, gingivitis, rhinitis, conjunctivitis, pneumonitis, enteritis, and
dermatitis. There may be clinical signs of neurologic dysfunction and concurrent
FeLV infection may aggravate the disease and result in neoplasia.
Diagnosis
 Clinical specimen: serum.
 Since cats infected with FIV remain infected for life. The detection of antibody to
FIV is considered to be the most reliable and convenient means of diagnosis.
Antibody may be detected by an immunofluorescence assay or by an ELISA.
ELISA test kits are available commercially and are considered to be highly
sensitive and specific.
Treatment
 Supportive care and antimicrobials for secondary infections.
 Vaccination for diseases other than rabies is not recommended.
 Reverse transcriptase inhibitors like those used in the treatment of HIV, e.g.,
azothiouridine, are claimed to reduce the severity of the disease. Interferon-α has
also been used.
Prevention
 A killed vaccine prepared from two isolates of FIV is commercially available in
the USA. Cats receiving the vaccine become serologically positive. Efficacy of
the vaccine is under current evaluation.
  Prevention is best accomplished by keeping cats indoors. Avoid contact with feral
and free-roaming cats.
 Only admit serologically negative cats to the household.

Bovine Immunodeficiency Virus Infection

Cause
Bovine immunodeficiency virus. The first isolation of bovine immunodeficiency
virus (BIV) was in 1972 from a dairy cow with persistent lymphocytosis in Louisiana.
Morphologically, the virus resembles the human immunodeficiency virus-1.
Additional isolations indicate that the virus is widespread in cattle; however, its
pathogenicity has not yet been demonstrated unequivocally, although there is some
evidence that it may be involved in predisposing cattle to other infections/diseases.
Experimentally infected cattle have exhibited lymphoid proliferation and decreased cell-
mediated cytotoxicity.
Jembrana Disease
The virus of this disease (Jembrana disease virus) resembles the bovine
immunodeficiency virus. The disease, which has only been reported as affecting Balinese
cattle (Bali, Indonesia), is characterized by a short course with fever, enlarged lymph
nodes and sometimes death. Cattle (Bos javanicus) that recover are viremic and thus
carriers.

Family: Reoviridae

Viruses in the family Reoviridae were originally isolated from respiratory and
enteric sources without any associated disease, namely orphan. (reo = respiratory, enteric,
orphan)

Properties
 Nonenveloped viruses with double or triple layered capsid and icosahedral
virion, diameter 60-80 nm
 Replicate in the cytoplasm
 Virions are composed of three concentric capsid layers, all with icosahedral
symmetry.
 The genome is composed of double stranded RNA, divided into 10-12
segments, total size 18-27 kbp: genus Orthoreovirus and Orbivirus, 10
segments, Rotavirus and Aquareovirus, 11 segments, Coltivirus, 12 segments.
They replicate conservatively in the cytoplasm, have a core-associated
transcriptase and some (especially orthoreoviruses) produce large cytoplasmic
perinuclear inclusions with a characteristic beehive pattern.
 Following entry into a host cell, the virion is partially uncoated. Unique to the
Reoviridae, replication occurs within partially uncoated virions as all of the
enzymes necessary for replication are present within the capsid. The dsRNA is
transcribed to form +sense RNA, some of which is sent into the cytoplasm for
translation by ribosomes. The remainder is packaged into partially assembled
virions. Within the virions, the complementary -sense RNA is synthesized
resulting in a dsRNA genome within the newly formed virions.
  Some serotypes share a common complement fixation antigen, but can be
distinguished by hemagglutination inhibition and neutralization techniques.
 They vary in stability; the orthoreoviruses and rotaviruses retain infectivity
over a wide pH range. Mammalian orthoreoviruses are resistant to a number of
common disinfectants.
 Genetic reassortment occurs between viruses within each genus or serogroup.
Classification
The reoviruses of vertebrates are placed in five genera and those of insects and plants in
four genera. The genera comprising the vertebrate viruses and their diseases are as
follows:
 Orbivirus (Have the capacity to replicate in some biting arthropods).
o Bluetongue
o African horse sickness
o Equine encephalosis
o Chuzan disease (Japan)
o Epizootic hemorrhagic disease of deer
o Ibaraki virus (A virus related to epizootic hemorrhagic disease virus of
deer. Causes an often severe disease of cattle in Japan, Taiwan and Bali.)
o Palyam virus [An orbivirus isolated from Culex species (India) and
Culicoides species (Australia and Nigeria) that causes arthropod-borne
disease characterized by fetal lesions and abortion].
 Rotavirus
o Rotavirus infections
 Orthoreovirus These viruses, which only infect vertebrates, are generally minor
causes of disease. They are spread by the fecal-oral and respiratory routes. Unlike
orbiviruses and rotaviruses they can be propagated in cell cultures with relative
ease. The mammalian orthoreovirus cause mild respiratory and enteric infections
in many animal species and humans.
Avian orthoreoviruses cause significant infections in chickens and turkeys.
 Coltivirus Members of this genus are mainly isolated from Ixodid ticks but also
from humans, deer and some small animals.
Colorado tick fever virus: A human disease occurring in the northwest U.S.;
acquired from the bite of infected tick; wild rodents probably serve as natural
reservoirs.
 Aquareovirus Species are recovered from sea and fresh water fish and some
marine invertebrates including clams and oysters.

African Horse Sickness Virus

Acute or subacute, febrile, arthropod borne disease of horses characterized by

oedema of subcutaneous tissues, lungs, haemorrhages of internal organs and
accumulation of serous fluids in the body cavities.

Host
Horses of all breeds are highly susceptible to natural infection. Mules and zebras
are less Affected and donkeys are generally resistant.
Distribution
The disease occurs principally in Southern, Eastern and Central Africa. From
there it spreads to Syria, Lebanon, Israel and other parts of middle east. It has been
recorded in Afghanistan, West Pakistan, Turkey and Iraq. It is a seasonal disease,
common in late summer in warm, humid, low lying marshy districts.

Properties
This virus is a double stranded RNA virus. The capsid is about 55 nm in diameter
with icosahedral symmetry. It is stable at pH 10. Rapidly inactivated at pH 3. Ether
resistant, deoxycholate resistant and trypsin resistant. Horse RBC's are agglutinated at pH
6.4 at 37°C for 2 hours.

Antigenic properties
Many serotypes have been described. In Africa 9 distinct serotypes are recognized
and considerable variation in virulence amongst individual strains within each
immunological type occurs. On the other hand only a single antigenic type involved in
the Asian outbreak in 1961.

Cultivation
The virus infects mice, rats, guinea pigs and other species of laboratory rodents by
intracerebral routes. Dogs can be infected by I/V or S/C infections. Ferrets can also be
infected by I/V route. Ferrets appear to be a particularly useful virus for isolation of
viruses from horses whose immunity has broken down during a natural exposure. Almost
all strains grow in the in the yolk Sac of fertile eggs. Virus can be propagated in cell
cultures of many different types. Monkey cell lines are most susceptible. In Rhesus
kidney cultures the characteristic CPE is rounding and shrinkage of infected cells which
remain attached to the glass.

Pathogenesis
After the bite of an infective arthropod, the virus replicates in the local lymphnode
before producing a transient viremia, which leads to infection of other tissues and organs
in the reticuloendothelial system and then a secondary viremia resulting in increased
vascular permeability, oedema, haemorrhage and intravascular coagulation.

Clinical signs

Incubation period is 6-9 days. 4 different clinical forms of illness were described.
i, Very mild form - only rise in temperature to 41°C.

ii. Acute or pulmonary form - (DUNKOP horse sickness) in this there is severe
dyspnoea, pyrexia and coughing with abundant frothy discharge from the nostrils. This
form is common in virulent outbreaks and mostly the affected horses die.

iii. Sub acute form: (DIKKOP horse sickness). It is characterized by remarkable

swelling of head, neck and supra orbital fossa associated with cardiac dyspnoea. This
form usually occurs when the immunity has been broken down by a natural infection or
in animals inoculated with mild strains of virus. This form is much milder than the acute
form and the affected animals recover.

iv. Mixed form: Combination of pulmonary form and cardiac form. Heart is
affected.

Mortality: In virulent form it is as high as 90% in horses and lower in mules. In milder
outbreaks mortality is 25%.

Lesions
It varies depending upon the severity of the case. In acute pulmonary form there is
extensive oedema of lungs and thorax may contain several litres of fluid. In more chronic
cardiac form there is oedematous infiltration of s/c tissue, sub- serosal tissue and other
tissues together with gross accumulation of fluid in the pericardial sac.

Immunity
Foals of immune dam are highly resistant for 5-6 months. Horses and mules
recovered from the natural outbreak are generally more resistant.

Transmission
It is not directly contagious. Various nocturnal biting insects including
mosquitoes are the most important vectors of infection in Africa. Culicoides species of
mosquito and insects belonging to Tabanidae family and genus stomoxys have been
incriminated in outbreaks in Turkey.

Diagnosis
Main method of transmission is by insects.
i. Clinical signs
ii. Post mortem findings.
iii. Blood from affected animal should be collected during early febrile phase of
the disease because the viraemic period is less. In dead animals spleen tissue can be
collected. The spleen tissue is injected into unweaned mice by the intracerebral route
and death of the mice occurs in 5-15 days of post inoculation. The infective mouse brain
is used as a source of antigen in the group specific CFT for primary identification.
Antigenic types can be found out by serum neutralization test or haemagglutination
inhibition test. Tissue culture techniques are now used for diagnosis of the disease.
Rhesus monkey cell lines are usually preferred.

Control
Vector control, quarantine of affected animals and vaccination are the main
methods of control.
Recently freeze dried live attenuated neurotropic mouse brain virus vaccine has
been proved useful and polyvalent vaccines from 7 or 8 antigenically distinct serotypes
appear to be safe and highly effective against all field strains. Regular annual
immunization is advocated. Tissue culture adapted virus, Hamster or Monkey kidney cell
culture vaccines are widely used in Africa and Middle East.

Blue Tongue Virus

Synonym: Sore mouth or sore muzzle, ovine catarrhal fever.

Acute arthropod borne virus, primarily affecting sheep characterized by catarrhal

inflammation of the mucous membrane of buccal mucosa and gastro intestinal tract. The
brain frequently involve the udder, coronary band of the foot and sensitive laminae of the
hoof. There is epithelial desquamation but no vesicle formation occurs.

Host
Sheep of all ages are highly susceptible but goats, cattle and wild ruminants
exhibit milder symptoms and may act as non-clinical carriers. Horses, dogs, cats, ferrets,
rabbits and guinea pigs are not susceptible to natural infection.

Distribution
Recognized in South, West and East Africa. Disease occurred in India in 1964.

Morphology
Virus particles are smaller of 53-60 nm in diameter. Viral capsid contains 32
capsomers. Mature virions are frequently surrounded by a thick outer envelope but
occasionally groups of 30-40 virus particles are observed in a membrane bound sac. The
nucleic acid component contains RNA and in early infections the RNA genome is double
stranded.

Physiochemical properties
It is much more resistant and remain viable in decomposed blood for many years.
Resistant to 20% diethyl ether. Survives dry in air and retain its infectivity in serum,
defibrinated blood, glycerol-oxalate phenol mixture for 25 years at room temperature. It
is stable at pH 5.6 to 8 and sensitive to acid pH. No haemagglutination property has been
recorded.

Cultivation
Can be readily isolated from infected sheep during febrile or early post febrile
periods. Growth is obtained in 6 days old embryonated eggs by yolk sac route and
temperature of incubation should be 33.5°C. It can be cultivated on CAM also. In CAM
mortality within 4-8 days with extensive hemorrhages of the developing embryos.
Intravenous inoculation of embryonated egg is 100 times sensitive than the yolk sac
route. Virulent strains have been attenuated without loss of antigenicity by serial passage
in eggs at temperature below 37.5°C. The virus is adapted to growth in brains of
unweaned mice and hamsters. The infected animals show signs of encephalitis and death
in 3-7 days. They can be propagated in primary cultures of calf kidney, adrenal and testis,
HELA cell cultures and also BHK21. CPE within 24-72 hours, which consists of foci of
enlarged and retractile cells.
Pathogenesis
The virus replicates initially in regional lymph nodes. It is then carried in blood or
lymph to other lymphoid tissues where further replication takes place. Virus localizes and
multiplies in the endothelium of small blood vessels producing vascular damage with
stasis, exudation and tissue hypoxia. The initiation and development of surface lesions in
areas of tissue hypoxia relate to minor trauma and may be complicated by secondary
bacterial infection.

Pathogenicity
Incubation period one week. Affected animals develop depression, anorexia,
pyrexia, copious salivation, development of excessive serous or mucous nasal discharge,
which dries up and forms a crust around the nose. Hyperemia, Petechiation and swelling
of the buccal mucosa, dental pads and tongue. Later the areas of hyperaernic regions
become "cyanotic" or "purplish blue". Multiple mucosal erosions of the mouth blood
stained diarrhoea occurs when the intestinal mucosa is involved which is always fatal.
There may be involvement of sensitive laminae and breaking of wool fibres. Heavy
mortality occurs in early stages and chronically affected animals become thin and
emaciated. Morbidity is 50%. The disease causes economic loss by way of loss of mutton
and wool. Abortion of pregnant ewes, which results in the loss of entire breeding season.
Young animals and Merino sheep are particularly susceptible.

Spread and transmission

The natural disease is transmitted by atleast one out of 21 species culicoides in
Africa and
Culicoides varypennis in USA. Reports of sheep led Melophagus ovinus harboring the
virus. Wild animal reservoir hosts play an important role in maintaining the infection
during the inter epizootic period.

Diagnosis
Clinical signs and lesions.
Samples suitable for virus isolation include unclotted blood from febrile animals
or fresh spleen and lymph node collected at post mortem. Confirmation of the disease is
by possible transmission to susceptible sheep or isolation of virus in unweaned mice or
growing in susceptible cell cultures or in embryonated eggs (i/v or yolk sac routes).
Virus can be identified by CFT. Agar gel diffusion test which is simple and
economic test can also be used. FAT can also be used.

Control
Line, attenuated, avianated vaccines are used as monovalent strain or
quadrovalent vaccine. Live vaccines should not be used in pregnant sheep. Sheep are
vaccinated annually, and several weeks before service and before the start of rainy
season. Lambs are not vaccinated until they are atleast 3 months of age.
 Equine Encephalosis
Equine encephalosis virus (seven serotypes) resembles closely the virus of AHS and
causes a frequently fatal disease that resembles closely AHS. The disease has been
reported from South Africa and is thought to be spread by biting arthropods and
particularly Culicoides species.
Chuzan Disease
The cause of this infection is a tentative species in the genus Orbivirus. The disease
occurs in southeast Asia and Japan and is characterized by congenital abnormalities in
calves including hydronencephaly and cerebellar hypoplasia. Development of these
abnormalities appears to be related to the age of the fetus at the time of infection, and is
thought to occur most often when the fetus is infected at about four months of age.
Affected calves may be blind and display seizures and opisthotonus. Beef cattle are more
often affected than dairy cattle.
Transmission is by Culicoides oxystoma.
Diagnosis is usually based on clinical history, microscopic lesions, and supportive
serologic results.

Epizootic Hemorrhagic Disease of Deer

Cause
Epizootic hemorrhagic disease (EHD) virus. There are eight serotypes of the EHD virus,
but only two of these (New Jersey and Alberta) occur in the United States.
Occurrence
The disease occurs in white-tailed and mule deer; other deer species are not susceptible.
Cattle, buffalo and non-white-tail deer are infected with various serotypes but do not
develop clinical disease.
Transmission
The virus is transmitted by the biting gnat, Culicoides variipennis.
Clinical & Pathologic Features
The disease may be subclinical peracute, acute, or chronic. The peracute form is
characterized by severe edema of the head and neck regions, including the tongue and
conjunctiva, and rapid death. Edematous lungs are noted at necropsy but there are few if
any signs of hemorrhage. Hemorrhages are more evident in animals that live longer
(acute form), and are usually present in the heart, rumen, and intestine. There are also
ulcerations on the tongue and dental pad and in the rumen and omasum.
The chronic form is characterized by lameness. Bluetongue virus may cause
similar disease manifestations. The virus of EHD may infect cattle, but most infections
are subclinical. One strain of EHD (Ibaraki) occurs in Southeast Asia and causes a
disease in cattle clinically similar to bluetongue.
Diagnosis
Clinical specimens: fresh heparinized blood, serum, and spleen.
Diagnosis of EHD is often made on the basis of clinical signs and lesions.
Confirmation requires isolation and identification of the virus. The virus can be
propagated in embryonated chicken eggs inoculated intravenously and in various
cell cultures, including the BHK-21 cell line.
Fluorescent antibody may be used to identify virus in frozen sections of affected
tissues.
 A high percentage of deer in the southeastern United States have antibodies to EHD
virus as determined by the agar gel immunodiffusion test.
Prevention
There are no practicable preventive measures.

Rotavirus

It causes diarrhoea in children and young animals like calves, lambs, piglets and
foals. Diarrhoea is referred to as "White scours" or "Milk scours". Isolates are divided
into seven antigenically distinct sero groups (A to G) based on reactions with the major
capsid protein, VP6. Most of the isolates belonging to serogroup A.

Epidemiology
Horizontal transmission occurs following ingestion of contaminated feed.
Because the virus is stable in the environment , premises may be heavily contaminated
and intensively reared animals are those most often affected.

Pathogenesis
The severity of infection is largely determined by the virulence of the infecting
viral strain, the amount of virus ingested and the level of maternally derived immunity.
The virus, which can survive gastric acidity, passes through the stomach and infects
enterocytes at the tips of villi in the small intestine. Because the rate of enterocyte
replacement is relatively slow in young animals, affected villi become stunted and
covered by cuboidal cells. these immature replacement cells have reduced levels of
disaccharides and defective glucose coupled sodium transport. Undigested lactose
provides an ideal substrate for bacterial proliferation in the intestinal lumen. In addition,
it exerts an osmotic effect, which results in the retention of fluid in the lumen and along
with impaired fluid absorption, contributes to the development of diarrhoea.

Clinical signs
Incubation period is 16 to 24 hours. Affected animals are anorectic and depressed and
produce light coloured, semi liquid or pasty faeces. In uncomplicated cases, animals
frequently recover within four days without treatment. Concurrent infection with other
enteric pathogens such as E.coli, Salmonella sp. and cryptosporidium sp. May add to the
severity of the diarrhoea and deaths may occur.

Diagnosis
Electronic microscopy is commonly used. Demonstration of virus in the feacal
sample by negative staining. Also by immunoelectron microscopy when there are few
numbers of viral particles. ELISA, PAGE (Polyacrylamide gel electrophoresis) can also
be used.
Tissue Culture
Initially non-cytopathogenic. After few (blind) passages, it becomes
cytopathogenic. Virus remains in the feaces for several months. So feed and water borne
transmission is possible.

Treatment
In mildly affected animals, water should be substituted for milk in the diet. Fluid
therapy by giving water alone instead of milk and antibiotics to control secondary
bacterial complications.

Prevention and control

Improvement in the hygienic practices, colostrum feeding which contain rotavirus
antibodies prevent infection. Inoculation of dam either with inactivated or attenuated
rotavirus vaccine will produce higher level of antibodies in colostrum and milk which
gives protection to calves.
Orthoreovirus
Avian Orthoreoviruses
These non-mammalian orthoreoviruses share a common group antigen. The eleven
serotypes produce syncytia in cell cultures and infections in chickens and turkeys. A
variety of infections are produced depending upon the particular orthoreovirus involved.
They include arthritis, tendinitis, gastroenteritis, hepatitis and myocarditis with weight
loss. The infections are common in commercial flocks.
Virus isolation and identification, although strait forward, is not usually feasible.
The viruses are readily identified by the fluorescent antibody staining of cryostat sections
of tissues.
Good management practices with thorough cleaning and disinfection of quarters
helps reduce losses. Active immunization is complicated by the involvement of different
serotypes of virus.

Mammalian Orthoreoviruses
These viruses, three serotypes, have been associated with mild respiratory and enteric
disease in many animal species. Disease may be severe if complicated by secondary
bacteria.

Family: Birnaviridae
Properties
 Nonenveloped icosahedral virion, 32 capsomers, 60 nm diameter.
 DsRNA genome, with two segments: A, 3.1 kbp;B, 2.9 kbp
 Four structural proteins, one nonstructural protein, virion transcriptase
 Cytoplasmic replication
 Survivors 60°C for 60 minutes, stable at pH 3-9.
 Member viruses occur in chickens (infectious bursal disease virus), fish
(infectious pancreatic necrosis virus), molluscs, and insects.

INFECTIOUS BURSAL DISEASE

Synonyms : Gumboro disease, Avian bursitis, Infectious nephritis.
Gumboro disease was recognized as a disease of chickens in 1957, and a virus
reported in Gumboro (USA) in 1962 by Coxgrove. In India first reported by Mohanty etal
(1971) in Uttar Pradesh.

Infectious bursal disease is most severe in chicks 3-6 weeks old, when the target
organ, the bursa of Fabricius, reaches its maximal stage of development. Naturally virus
has special affinity for pre-B lymphocytes of the bursa of Fabricius, leading to acquired
B-lymphocyte deficiency in affected birds.

Properties
Virus is inhibited by formalin and not by ether, chloroform and phenol etc. It is
stable at pH-2. but inactivated at pH 12. There are three serotypes, 9 polypeptides - small,
medium and large.

Cultivation
Infectious bursal disease virus (IBD) replicates in both chicken and mammalian
cells. Chicken embryo kidney cell culture is good for isolation. Cytopathogenic effect
seen in 3-5 days. Chicken embryo fibroblasts are more sensitive than fertile eggs for viral
titration.

Pathogenesis
The most striking feature of the pathogenesis and pathology is the selective
replication of infectious bursal disease virus in the bursa of Fabricius, which is enlarged
(up to five times its normal size), edematous, hyperemic, and cream- colored, with
prominent longitudinal striations.
Hemorrhages occur beneath the serosa, and there are necrotic foci throughout the bursal
parenchyma.
The virus is found in the gizzard, duodenum and kidney 12 hrs post infection
(parenteral route or natural infection) and in the spleen, bursa and kidney in 24 hrs after
oral infection. The virus replicates in the macrophages of the necrotic follicles of bursa
and also in the lymphocytes and highest virus concentration after 4-8 days post infection.
Low concentration of virus in the spleen and thymus.

Lesions
Initially dehydration, haemorrhages in the leg and thigh muscles, hepatic infarcts,
enlarged kidneys with pronounced tubules. Ureters are filled with urates. Bursa highly
enlarged twice or 2-4 times its normal size-approximately three days after infection and
becomes edematous and haemorrhagic. Atrophies by 12 days, post infection. Spleen
slightly enlarged and mottled with deep red spots. Haemorrhagic lesions present in the
proventriculus and hypertrophy of the liver is noticed.

Haemotological changes
Low blood calcium level, lymphopenia, hypertonic dehydration and related
erythrocytosis. 1-7 days after infection reduction in the number of erythrocytes. Total
leucocyte count decreases two days post infection, increases after 4 days and is in peak 7
days post infection. Differential count - low percentage of lymphocytes with a high
degree of heterophils. Transient delay in the coagulation time.

Immunosuppressive properties
IBD infection reduces the HI antibody litres and lgG levels of Newcastle disease
vaccination. Further humoral antibody response to various vaccines was also suppressed.
Immunosuppressive property as been recorded in the cases of Infectious bronchitis (IB)
and Infectious laryngeotracheitis (ILT) also. Report of suppression of CMI by IBD also
has been recorded.

Diagnosis
Age, History of the bird, clinical signs, mortality pattern, gross and
histopathological lesions. Immunofluorescence of impression smears of bursal tissue, gel
diffusion tests with infected bursal tissue as the antigen, electron microscopy of bursal
specimens, and viral isolation in embryonated eggs are all useful in confirming the
clinical diagnosis. The presence of virus or viral antigen can be detected in bursal tissue
by immunofluorescence for 3-4 days after infection, for 5-6 days by immunodiffusion,
and for up to 14 days by viral isolation.
Neutralization tests and ELISA are reliable methods for serodiagnosis.

Prevention and control

Maternal antibodies protect day old chicks against death and clinical IBD.
By vaccination.
Ideal age of vaccination –14 to 16 days of age by using IV95/ Intermediate plus/
MB strain and this vaccine should be repeated after one week.

Two methods of control

I. Vaccination of breeders to produce high maternal antibody level in the offspring
to prevent infection during first week of age.

2. Vaccination of the progeny at day old to prevent early infection.

Long lasting and more uniform levels of maternal antibody was present in,
the chicks which were from breeders receiving two live IBD vaccines and are
inactivated oil emulsion IBD vaccine during the growing period.

ORDER: MONO NEGA VIRALES

The order mononegavirales encompassing four families: paramyxo viridae,

rhabdo viridae, filo viridae and borna viridae. All the member viruses of these families
are enveloped, covered with peplomers and all have genomes consisting of a single
molecule of negative sense, single stranded RNA.

Family: Paramyxoviridae
 Paramyxo and Orthomyxo were originally under myxoviruses because of the
morphologic similarity of the virions and the fact that the prototype viruses, Newcastle
disease virus and influenza virus, each carry a hemagglutinin and a neuraminidase.
However, it was later realized that the viruses of each group differ in such basic
properties as genome structure and mode of replication; hence they were separated into
two families.
The family paramyxo viridae is subdivided into the sub families paramyxovirinae
and pneumovirinae.

Paramyxo viruses of animals

Subfamily: Paramyxo virinae

Genus Virus Disease
Respiro virus Bovine para influenza virus 3 Respiratory disease in cattle and sheep
Murine parainfluenza virus 1 Severe respiratory disease in laboratory rats
(Sendai virus) and mice
Avula virus Newcastle Disease virus Severe genralised disease with CNS signs in
(Avian paramyxo virus –1) domestic and wild fowl
Avian paramyxo viruses 2-9 Respiratory disease in fowl
Canine parainfluenza virus 2 Respiratory disease in dogs
Rubula virus Porcine rubula virus Encephalitis, reproductive failures, corneal
opacities in swine
Mumps virus Parotitis in humans
Rinderpest virus Severe generalized disease in cattle and
wild rumiants
Peste des petits ruminants virus Severe generalized disease in sheep and
goats
Morbilli virus Canine distemper virus Severe generalized disease with CNS signs
in dogs and members of felidae, mustelidae
family
Measles virus Severe systemic disease with respiratory
and CNS signs
Equine morbilli virus Acute respiratory disease syndrome in
horses and humans.

Subfamily Pneumo virinae:

Genus Virus Disease
Pneumo virus Bovine respiratory Respiratory syncytial disease in cattle and
syncytial virus sheep
Meta pneumo virus Turkey rhino tracheitis Severe respiratory disease in turkeys,
virus swollen head syndrome of chickens.

Properties
Paramyxo virus virions are pleomorphic in shape (Spherical as well as
filamentous form), 150-300 nm in d.m. They are enveloped, covered with large
peplomers (8-20 nm in length), and contain a herringbone shaped helically symmetrical
nucleocapsid. The genome consists of a linear, minus sense ssRNA genome, 15 to 16 kb,
encoding 10-12 polypeptides, including NP (or N), P, M, F, L, and HN (or H or G),
which are common to all genera.

Both HN (Haemagglutinin- neuraminidase) and F (fusion protein) play key roles

in pathogenesis of all paramyxovirus infections. Cell attachement is mediated via the HN
protein. This glycoprotein elicits neutralizing antibodies that inhibit adsorption of virus to
cellular receptors. The fusion protein is present on newly formed virions in an inactive
precursor form that is activated by proteolytic clevage by a cellular protease. After
cleavage, the newly generated amino-terminal sequence is involved directly in
fusion.when a host cell does not contain appropriate proteases the virus formed is not
infectious. Because the fusion protein is essential for direct cell-to-cell spread via fusion,
it plays a key role in the pathogenesis of paramyxo virus infections, including perisitent
infections.

Functions of viral proteins

Function Respiro virus and Morbilli virus Pneumo virus
Avula and Rubula
virus
Attachement protein: HN H G
haemagglutinin, induction of (No Haemagglutinating
productive immunity activity)
Neuraminidase: virion release, HN None None
destruction of mucin inhibitors
Fusion protein: cell fusion, virus F F F
penetration, cell-cell spread,
contribution to induction of
protective immunity
Nucleoprotein: Protection of NP N N
genome RNA
Transcriptase: RNA genome L and P L and P L and P
transcription
Matrix protein: Virion stability M M M

Paramyxo viruses replicate within the cytoplasm. Virons attach via HN protein to
cellular sialoglyco protein or glyco lipid receptors. The F protein then mediates fusion of
the viral envelope with the plasma membrane at physiologic pH. After replication, the
virus will bud from the plasma membrane. In infected cells, the virus produces
characteristic syncytium formation with intracytoplasmic or intranuclear (morbillivirus)
inclusion bodies. Most strains agglutinate avian and certain species of mammalian RBCs.
 Three genera: Paramyovirus, Morbillivirus, Pneumovirus

NEWCASTLE DISEASE
(Avian Paramyxovirus 1)

Synonym: Ranikhet disease, Avian Pneumoencephalitis, Pseudo fowl pest, avian pest,
 Avian distemper, Vellaikalichal (Vernacular), Doyle's disease

History
Newcastle is a highly contagious viral disease of birds. It was first observed in
1926, in Java, Indonesia and in Newcastle-upon-Tyne, England. The name Newcastle
disease was coined by Doyle. In India in July 1927, the virus was recognized in Ranikhet
by Edwards. Hence it named as Newcastle disease or Ranikhet disease. Nine serotypes of
avian paramyxoviruses but only avian paramyxovirus 1 is associated with clearly defined
disease.

Etiology
Order: Mononega virales
Family : Paramyxo viridae
Subfamily : Paramyxo virinae
Genus: Avula virus
Species : Avian paramyxo virus type -1
Morphology
Single molecule of Single stranded negative sense RNA viruses. They possess
helical symmetry. A herringbone nucleocapsid, about 18nm in d.m. is seen. They are
enveloped and it has number of surface projections.

The genome of NDV codes for six proteins. ( 3’ N-P- M-F-HN-L-5’ )

NP – Nucleocapsid protein
P – Phosphorylated - Nucleocapsid associated
M – Matrix protein
F – fusion protein, forms the smaller of the surface projections.
HN – haemagglutinin and neuraminidase – larger of the surface projections
L - RNA directed RNA polymerase associated with the nucleocapsid.

The RNA together with NP, P, and L proteins forms the ribonucleoprotein
complex, which serves as a template for RNA synthesis. HN is responsible for
attachment and release of virus from the host cell in addition to its serological
identification. F play a major role in fusion of the virus and cell membrane and it has a
critical role in pathogenesis. NDV causes agglutination of chicken and human O
erythrocytes. The agglutination reaction is not permanent and after 30 minutes as a result
of loss of receptor the agglutination is lost due to the action of neuraminidase and this
phenomenon is referred as elution.
Only one serotype. But many strains are there. All strains are immunologically
same. Some of the important virus strains are as follows
Pathotype Strains
Velogenic Hertz 33, Texas GB
Mesogenic K, R2B
Lentogenic F, Lasota, Ulster 2C
Avirulent V4

Resistance
 The virus is comparatively stable virus and survives for long periods at ambient
temperature, especially in faeces and can persist in houses (in faeces, dust etc.) for upto
12 months. However it is sensitive to disinfectants, fumigants and sunlight. The virus is
inactivated by temperatures of 560C for 3 hours or 600 C for 30 min, acid pH, formalin
and phenol, and is ether sensitive.

Cultivation

NDV can be readily cultivated in embryonated eggs. The preferred route is

allantoic cavity in 9-11 days old embryos. Virulent strains cause death of embryos in 24
hrs with haemorrhages through out the body particularly in occipital region. Lentogenic
strains do not cause death. Presence of virus is usually confirmed by HA and HI test
using specific serum.

The virus also grows readily in primary cell cultures like chicken embryo
fibroblast and pig kidney cells and in BHK 21 cell line.

Epidemiology
In addition to chickens, turkeys, pheasants, guinea fowl, ducks, geese, and
pigeons, a wide range of captive and free ranging semi domestic and free-living birds,
including migratory waterfowl, is susceptible. NDV has been reported at least 241
species of birds. In birds that survive, virus is shed in all secretions and excretions for at
least 4 weeks.

Transmission occurs by direct contact between birds by the airborne route via
aerosols and dust particles by inhalation, ingestion of feaces or contaminated material, by
inanimate objects, by movement of people, contaminated feed and water, by non-avian
species like flying insects, fly (Musca domestica), ectoparasites (Argus persicus),
endoparasites (Ascardia galli, Eimeria tenella etc.) and by vaccine also (incomplete
inactivation and exaltation).

With lentogenic strains, transovarian/vertical transmission is important, and virus

–infected chicks may hatch from virus-containing eggs. Some psittacine speciesmay
become persistently infected with velogenic virus and excrete it intermittently for more
than 1 year without showing clinical signs. Virus may also be disseminated by frozen
chickens, uncooked kitchen refuse, foodstuffs, bedding, manure and transport containers.

Pathogenesis
It mainly depends upon the virulence and tissue tropism of the virus. Other factors
are age of the birds, immune status, route of exposure, magnitude and duration of the
infecting dose, susceptibility of the host species, external factors like stress and
temperature.
 Strains of NDV differ widely in virulence, depending on the cleavability and
activation of their hemaggutinin – neuraminidase and fusion glycoproteins. Avirulent
strain produce inactive precursor proteins; in virulent strains these precursors are cleaved
and activated readily. Cleavage activation must occur in the same tissues in which intial
viral replication occurs if the infection is to be progressive – the presence of cellular
proteases as well as the strain-based cleavability of the viral glycoprotein precursors
defines the tissue tropism of various viral strains and their capacity to spread rapidly.

Virus replicates at the site of entry like conjunctiva, respiratory and alimentary
tracts. Destruction of mucus, which rapidly spreads to other ciliated goblet cells and
acini. Then viremia results with multiplication of virus in many organ's like spleen, lungs,
kidney and bursa. Then multiplies in duodenum, trachea, pancreas and brain. The virus is
grouped into five pathotypes on the basis of the predominant signs in affected signs.

They are as follows

Viscerotropic velogenic (Doyle's form or Asiatic form): Very acute form with
hemorrhagic lesions of the digestive tract. The infection caused by this also
called as Asiatic or exotic ND.
Neurotropic velogenic (Beach's form): Viruses causing disease characterized
by high mortality, which follows neurological and respiratory disease.
Mesogenic (Beaudette's form): Acute respiratory and some times lethal
nervous infections of young chicks.
Lentogenic (Hitchner's form): Mild or inapparent respiratory infections of
chicks.
Asymptomatic enteric: Inapparent intestinal infection

Clinical features
Incubation period is varies from 2 to 15 days (V V ND – 2 to 6 days). Morbidity
is usually high and mortality is upto 100%. In chickens respiratory, circulatory, gastro
intestinal, and nervous signs are seen. A combination of dyspnoea, cyanosis of comb and
wattles, and clonic muscular spasm is indicative.

There is a loss of appetite, listlessness, abnormal thirst, huddling, weakness, and

somnolence. Intestinal symptoms may include crop dilatation, presence of foamy mucus
and fibrinous exudates in the pharynx, a similar discharge from the beak, and yellow –
green diarhoea. Nervous system involvement is indicated by paralysis of wings and /or
legs, torticolis, ataxia or circular movements, bobbing-and-weaving movements of the
head, and clonic spasms. In layers, there is a sudden decrease in egg production together
with depigmentation and/or loss of shell and reduction in the albumen quality of eggs.
In VVND, edema of the head, especially around the eyes may become apparent
after birds have been sick for 2 or 3 days. This odema usually does not involve comb and
wattle as in a case of High pathogenic Avian Influenza. A dark ring sometimes forms
around the eye, probably due to cyanosis and poor blood cicculation in the odematous
tissue. This ‘black eye’ appearance is especially visible in white chickens. Bile stained,
greenish-dark diarrhoea may be noted 2 to 3 days after onset of illness.
Lesions
Gross pathological findings include ecchymotic hemorrhages in the larynx,
trachea, esophagus and throughout the intestine. The most prominent histological lesions
are necrotic foci in the intestinal mucosa and lymphatic tissue and hyperemic changes in
most organs, including the brain. Non-purulent encephalomyelitis in CNS is also seen.

Virulent velogenic strains cause predominantly haemorrhagic lesions, in

particular at the esophagus/proventriculus and proventriculus/ gizzard junctions and in
the posterior half of the duodenum, the jejunum, and ileum. These lesions are virtually
pathognomic for veogenic strains. Haemorrhages with necrosis in the intestinal wall. In
addition to this egg peritoinitis, edematous, haemorhagic or degenerated ovary are also
seen.

Diagnosis
Samples to be collected
Cloacal swabs from live birds. From ailing bird’s oro-nasal swabs, brain, bile,
caecal tonsils, liver, spleen, lung, kidney and heart tissue.

Based on history and clinical signs.

Based on iIsolation and identificaton of virus

Inoculate suscpected materials into 4-6 week old SPF chickens either by cloacal,
conjunctival or respiratory route. If VVND is present, the inoculated chickens usually die
in 3 to 7 days. Which reveals typical visceral lesions on PM examination. Neurotropic
viruses cause severe neurologic and respiratory signs.

Confirmation can be done by isolate the virus in 9-11 day old embryonated eggs
with spleen, brain and lungs and demonstrate the virus by HA and HI test.

Pathotyping of ND
NDV are classified into velogenic, mesogenic and lentogenic based on following
three tests

Intra cerebral Pathogenicity index (ICPI)

Inoculte clarified suspected materials or fresh infective allantoic fluid into 10-day-
old SPF chicks. Each bird is examined at 24 hr intervals for eight days and graded zero if
normal, one if sick and two if dead. The index is the mean score per bird per observation
over the 8-day period. The most virulent viruses give ICPI values approaching the
maximum score of 2.0, while lentogenic viruses give values of, or close to, 0.0.

Intra venous Pathogenicity index (IVPI)

Inject 0.1 ml of the diluted virus intra venously into each of ten 6-week-old SPF
chickens. Birds are examined at 24 hr intervals for 10 days and scored at each
observation –0 if normal, 1 if sick, 2 if paralysed or showing other nervous signs, and 3 if
dead. The IVPI is the mean score per bird per observation over the 10-day period.
Lentogenic and mesogenic strains have IVPI values of 0, whereas the indices for virulent
strains will approach 3.0.

Mean Death time (MDT)

The MDT in embryonated eggs means, time in hours for the minimum lethal dose
to kill all the inoulated embryos. The MDT has been used to classify ND virus strains
into velogenic (taking under 60 hrs to kill); mesogenic (taking 60-90 hrs to kill); and
lentogenic (taking more than 90hrs to kill).

Demonstration of viral antigen on impression smears by IFT and other serological

tests viz: HI, ELISA and molecular techniques like RT-PCR are highly valuable.

Immunity
Antibody production is rapid. Haemagglutination- inhibiting antibody can be
detected within 4 to 6 days of infection and persists for atleast 2 years. The level of
hemagglutinating-inhibiting antibody is a measure of immunity. Maternal antibodies
protect chicks for 3 to 4 weeks after hatching. lgG confined to the circulation does not
prevent respiratory infection but blocks viremia; locally produced IgA antibodies play an
important role in protection in both the respiratory tract and the intestine.

Prevention and control

Control is mainly by vaccination. There are three types of vaccines used for ND:
Live lentogenic-F, La Sota.
Live mesogenic-Komarov, Roakin, Mukhteswar, R2B.
and inactivate vaccines.

Live lentogenic vaccines are usually derived from field viruses that have been
show to have low pathogenicity for poultry but produce an adequate immune response.
Typical vaccine strains are HB1, La Sota, F, V4 or Ulster 2C viruses. These vaccines are
given to young chicks through eyes or nostrils. They are very safe and only La sota will
produce mild post vaccinal reaction in certain flocks. This type immunization is also
called as priming.

Mesogenic strains are used only in areas where ND is epidemic and in village
chickens. Normally mesogenic chickens, such as Komarov, Roakin, Mukhteswar and
R2B are used as secondary vaccines after a primary vaccination with a lentogenic
vaccine.

Inactivated vaccines are produced by growing a ND virus in eggs, and then trating
the infective allantoic fluid with an inactivating agent and adjuvant, such as formalin or
betapropiolactone-and mineral oil as adjuvant. Inactivated vaccines are usually applied
after an initial priming vaccnation with a live vaccine.

Thermostable, recombinant ad marker vaccines are also available.

 Vaccination is usully applied through water, aeosol spray, eye or nasal drops, or
by injection.
Control
i. National Level:
Quarantine, slaughter and vaccination (Ring vaccination)
ii. Farm level:
Strict hygienic practices and medical measures
By vaccination.
Import restrictions on chickens and eggs
Quarantine of psittacine birds in the same air space as non-immune chickens.
Slaughter policy once diagnosed. Followed by thorough disinfections.
Vaccination with lentogenic or inactivated virus in countries where velogenic
virus is endemic. Broilers are vaccinated at 7, and 21 days. Layers are also vaccinated at
10 and 22 weeks (point of lay). In addition to vaccination usual biosecurity measures
should be followed.

Rinderpest

Svnonvm : Cattle plague

Rinderpest is a highly infectious acute or subacute systemic disease of cattle,

which is characterized by necrosis, and erosions of the mucosa in the respiratory and
digestive tracts. Early constipation, usually preceded by dehydration and prostration, is
followed by diarrhea.

Properties
There is only one serotype, which is antigenically stable and exhibits extensive
cross reactivity with the other morbilli viruses. Virus is labile and is rapidly inactivated in
decaying carcasses, within a few hours under tropical conditions. In manure the virus
remains infectious for about 48 hours, whereas meat, spleen and lymph nodes at 5°C
remain infectious for 2 to 3 days. Half life is 1-3 hours at 37°C and only minutes at 56°C.
The infectivity losses within a few months at 4°C and remains at -70°C. After one year a
fall in titre of the virus occurs. It is best preserved by lyophilisation or by addition of 2%
DMSO (Dimethyl Sulphoxide - when stored at 4°C or lower temperature). Strong alkalies
are the best disinfectants and glycerol, phenol, formalin, a-propiolacton destroy the
infectivity quickly (but not antigenicity). Anti rinderpest sera inhibits measles
haemagglutinin activity (MHl).

Cultivation
The virus grows in sheep, goats, rabbits, hamster and white mice. It can be grown
in chick embryos by intravenous or CAM route. Cytopathogenic effect (CPE) in
susceptible cells from cattle, sheep, goats, chick embryos, pigs, hamsters and man. CPE
consists of large, well defined multinucleated giant cells known as "syncytia". Infected
cell cultures have stellate or spindle shaped cells with large fine anastamosing
intracellular processes. The virus multiplies in the cytoplasm of the host cells like
verocells (African green monkey kidney cells). Both intra cytoplasmic and intranuclear
inclusion bodies are present.

Pathogenicity
The normal route of infection is through nasopharyngeal mucosa. The course of
the disease comprises of 4 stages.
I stage
Incubation period: 2-9 days. It depends according to the strain and dose of the virus.
The virus multiplies rapidly in the lymphoid tissue, lungs, bone marrow and intestines.
Active proliferation of the virus in the tissue results in fever.

II stage
Prodromal phase: There is first rise in temperature - 105-107°C (41-42°C) and lasts for
about 3-5 days until the appearance of lesions in the mouth. Animal show depression,
restlessness and anorexia. Muzzle is dry, starry coat and initial constipation noticed,
Leucopenia with onset of fever and persists till death.

III stage
Mucosal phase: Mouth lesions on the inner lips and adjacent gums. Visible mucous
membrane are congested. Greyish foci with necrotic centres, and shallow erosions with
bleeding. Ulcers with bran like deposits noticed. Smacking as in FMD is not common.
Animal is restless and shows excess thirst. Temperature is high and recedes. After that
diarrhoea begins. Rapid dehydration, marked weakness and severe progressive
emaciation leads to death.

IV stage
Convalescent phase: Mouth lesions start healing. Rapid regeneration of the affected
epithelium noticed. Slow recuperation of general health.
Mortality in cattle, sheep and goats and pigs is 90%. In India "Hill Zebu cattle"
are more susceptible than "Plain zebu cattle". Wild game are often affected.

Pathology
Inflammatory changes noticed in the mucosa of the abomasum and intestine.
Streaks of haemorrhages in the intestine - caecum, colon and rectum, mimicking "zebra
markings" (Linear streaks of haemorrhages).

Diagnosis
 Clinical symptoms
 Isolation and identification of the virus
 Detection of rinderpest specific antigen
 Demonstration of specific rinderpest antibodies
 Collection of defibrinated blood at the peak of the temperature
 Paired sera sample
 Spleen and mesentric lymphnodes from dead animals.
Differential diagnosis
 Haemorrhagic septicemia, FMD, Mucosal disease and Bovine Viral Diarrhoea.

Immunity: Recovered animals are life long immune.

Prevention and control

Vaccination
i. Lapinised vaccine: Immunity last for 1-7 years;
ii. Goat tissue vaccine: Immunity last for 13 years or life long;
iii. Tissue Culture Rinderpest vaccine (TCRV): It is very effective and useful in
outbreak areas (Calf kidney cell culture vaccine, verocell line vaccine).

CANINE DISTEMPER

Synonym: Hardpad disease, Carre’s disease.

It is a highly infectious, acute or subacute, incurable, often fatal, multi systemic

febrile disease of dogs and other carnivores, which has been known since 1760. Edward
Jenner first described the course and clinical features of the disease in 1809; its viral
etiology was demonstrated by Carre in 1909.

Host
The host range of CDV embraces all species of the families Canidae (dog, fox,
coyote, jackal, wolf), Procyonidae (raccoon, panda), Mustelidae (ferret, mink, skunk),
and the large numbers of the family Felidae (lions, leopards, cheetahs, tigers)

Morphology

CDV has a spherical shape and diameter ranging from 100 to 250nm. It contains
negative sense, single stranded, non-segmented RNA. Virions contain a herringbone-
shaped helically symmetrical nucleocapsid. Virions are enveloped. The envelope contains
two viral glycoproteins and one or two nonglycosylated proteins. Genomes encoding six
proteins, which includes, NP (or N), P - Phospho protein, M – Matrix Protein, F- Fusion
Protein, L – Polymerase protein and H – Haemagglutinin attachment protein. The virus
replicates in the cytoplasm and buds from the plasma membrane.

There is only one serotype of the virus, but strains vary in virulence.

Resistance
Inactivation is rapid at 37oC and occurs after a few hours at room temperature.
Disinfectants readily destroy viral infectivity. However, at normal temperatures the virus
is very stable and can stay active in infected materials for several weeks, provided the
materials are not exposed to sunlight. At below zero temperatures the virus can survive
for many months.
Cultivation
Cell culture
 CDV grows very well in MDCK, Vero, Primary dog lung cells, mitogen
stimulated canine or ferret lymphocytes. Typical cytopathic effects such as the formation
of stellate cells and syncytia can be observed 3 to 12 days after infection. Syncytium
formation, acidophilic intracytoplasmic and intra nuclear inclusion bodies are
characteristic.

Embryonated eggs
The virus also grows in the chorioallantoic membrane. However, no changes are
produced upto 10th passage. Subsequent passages show increased thickening and whitish
areas in the CAM.

Laboratory animals
The distemper viruses are cultivated in live dogs or ferrets. Few strains also grow
in suckling mouse.

Transmission

The virus is shed in all secretions and excretions from the fifth day after infection,
which is before the onset of clinical signs, and continues, sometimes for weeks. Dogs in
recovery may continue to shed the virus for several weeks after symptoms disappear and
act as source for contamination. Transmission is mainly via direct contact, droplet and
aerosol. Because an intact fatty envelope is required for infection, virus transmission
must involve dog-to-dog contact or at least contact with extremely fresh (virus able to
survive within droplet for 30min) infected body secretions. Young dogs are mainly
susceptible than older dogs, with the highest susceptibility being between 4 and 6 months
of age, after puppies have lost their maternal antibody.

Pathogenesis

The first pathogenic event is the local replication of the virus for 2 to 4 days in
cells of the upper respiratory tract or in conjunctival epithelium. After further
multiplication in regional lymph nodes, the virus enters the bloodstream, carried within
lymphocytes, to produce a primary viremia (synchronous with the first bout of fever) that
spreads the virus to the reticulo endothelial system. The effects of viral replication at
these sites are manifested by hyperplasia and by the presence of multinucleated giant
cells in lymphoid organs. Virions formed in these sites are carried by lymphocytes and
monocytes to produce a secondary viremia, coincide with second peak of fever.

The rate of spread and distribution of virus after days 8 and 9 varies depends on
the rate of development of neutralizing antibody. If the antibody titre reaches 100 or
higher, the virus disappears rapidly from the lymphatic tissues and the infection remain
sub clinical. If measurable antibody is not present the virus spreads throughout the body.
Extensive infection of epithelium in the intestinal, respiratory, and urogenital tracts, skin,
and exocrine and endocrine glands occurs, as well as continued infection of mononuclear
cells in the lymphoreticular system.
Clinical features
Incubation period is 3-7 days. CD occurs in several forms. In the rare peracute
form of the disease there is sudden onset of fever and sudden death.
In the most common acute form of the disease, after an incubation period of 3-6 days,
infected dogs develops a biphasic rise of temperature up to 410C, with the second peak
corresponding to the onset of severe leukopenia and other clinical signs. After the fever
gastrointestinal and respiratory symptoms are more common. Which includes
catarrhal inflammation of the larynx, bronchi, tonsillitis, cough, severe vomiting and
watery diarrhoea. After the onset of disease, CNS signs may occur: behavioral changes,
forced movements, local myoclony, tonic-clonic spasms, epileptic –like attacks, ataxia,
and paresis. Pustules resemble allergic eruptions. Occasionally pustular eruptions occur
on the abdomen and on inner surface of the legs.

The duration of disease varies, depending on complications caused by secondary

bacterial infections.The subacute neurological form is characterized by frank
encephalitis with convulsions and seizures and most surviving dogs have permanent CNS
sequelae such as nervous ticks or involuntary leg movements.

Finally, two late forms, old dog encephalitis (slowly progressive loss of
neurologic functions) and hard-pad disease (hyperkeratosis of foot pads and the nose)
are seen in old dogs. Neurological signs include
1) Chorea - a peculiar pattern of twitching of ears
2). Localized twitching of muscle or group of muscles
3). Paralysis (beginning in the hind limbs followed by ascending paresis)
4). Convulsions – characterized by salivation and chewing gum movement (Petit mal)
_ frequent and severe seizures characterized by dogs fall on its sides
and paddle its legs, with involuntary urination and defecation.
Both of these late diseases may occur together and both may end in death. In CD
the mortality ranges from 30 to 80%

Diagnosis
Samples to be collected
Live animals
Lymphocytes, Serous discharge, discharges from all secretions and excretions,
serum, impression smear from cornea, conjunctiva and from pustules
Dead animals
Lung, stomach, bladder and intestinal tissue. Trachea, salivary glands, adrenal
glands, lymph nodes, spleen and skin.

Diagnosis is based on
i. Classical clinical signs
ii. Demonstration of distemper inclusion bodies:
The inclusion bodies, which red stained (eosinophilic) oval structures are readily
visible in urinary bladder tissues, conjunctival impression smears, epithelial cells of the
salivary glands, adrenal glands, lymphnode, spleen and skin.
 iii. Virus isolation and identification in cell cultures and in laboratory animals
iv). Demonstration of viral antigen by FAT of the conjunctival impression smears of the
live animals and impression smears of lung, stomach, intestine, liver and bladder tissue of
the dead animals.
v). By serology
SNT and Complement Fixation Test
Cerebrospinal fluid antibody levels - Distemper antibodies in CSF are highly
indicative of distemper infection as vaccine induced antibodies do not cross the blood
brain barrier into the CSF fluid.

Differential diagnosis
Canine hepatitis (Rubarth's disease - adeno virus)
Canine parvovirus disease
Leptospirosis
Toxoplasmosis
Rabies

Prevention and control

Control is based on adequate diagnosis, quarantine, sanitation, and vaccination.
The virus is extremely fragile, and disinfection of premises is achieved easily with any
standard disinfectant. There are no antiviral drugs to treat CD. Hyper immune serum can
be used to the affected animals immediately after exposure.

Vaccines
Tissue culture vaccine, egg adapted vaccine, modified live virus vaccines are
available. Successful vaccination is depends on the absence of interfering maternal
antibody. Pups can be vaccinated at the age of 8 weeks and again at 12-16 weeks.
Revaccination is necessary. Combined vaccine - ICH and CD or ICH, CD and
leptospirosis are also available. Human measles virus ( because antigenically related) can
also be used to vaccinate dogs against CDV, but measles virus doe not produce long
lasting immunity.

First vaccine against CD was produced by Laidlaw and Dunkin in 1928 with
inactivated spleen tissue. After this Distemperoid virus vaccine was discovered by
Green in 1939. Distemperoid virus produced by passaging in ferrets or minks. A strain of
CD virus by continuous passages in ferrets and minks got adapted which is highly
pathogenic for ferrets and minks but of reduced virulence to dogs and foxes. It stimulates
a higher degree of active immunity in dogs. It interferes with the field CD virus. Haig in
1945 adapted this Green's distemperoid virus in eggs by CAM route for vaccine
production. Recently vero cell culture adapted vaccines are produced.

In IVPM, egg (CAM route) adapted Lederle strain of CD virus vaccine is

available in freeze-dried form. Indication for this vaccine is
For pups of 12 weeks of age or above single dose of vaccine is sufficient.
For pups below 12 weeks of age 2 doses of vaccine should be administered. First
dose after weaning ( 8-10 weeks) and second dose at 12 –16 weeks.
 PESTE DES PETITS RUMINANTS (PPR)

Synonym: Goat Plague, Kata

PPR is a highly contagious, acute or subacute viral disease of goats and sheep
characterized by fever, erosive stomatitis, conjunctivitis, gastro enteritis and pneumonia.
PPR was first described in 1942 in West Africa and it is closely related to rinderpest
virus, CDV, equine influenza and human measles virus.

Host
PPR is primarily a disease of sheep and goats. Goats, particularly young ones,
(4 months to 1year of age) are usually more severly affected than sheep. The infection
also occurs in wild ungulates. Catte, buffaloes, camels and pigs are rarely susceptible.
They do not exhibit clnical signs and are unable to transmit the disease to other animals.
Sahalian breeds of goat and sheep are resistant to PPR.

Morphology

PPRV is pleomorphic. The virus measures approximately 150nm in diameter. It

contains negative sense, single stranded, non-segmented RNA. Virions contain a
herringbone-shaped helically symmetrical nucleocapsid. Virions are enveloped. The
envelopes are covered with minute projections, which are the surface glycoproteins (H
and F proteins) responsible for cell attachment and fusion. Genomes encoding six
structural proteins, which includes, NP (or N), P - Phospho protein, M – Matrix Protein,
F- Fusion Protein, L – Polymerase protein and H – Haemagglutinin attachment protein
and two non-structural proteins, which includes, C and V. The virus replicates in the
cytoplasm and buds from the plasma membrane. PPRV caused haemagglutination of
chicken RBC. However, the pH of PBS should be at 6.8

PPRV is differentiated from rinderpest virus through nucleic acid probes for N
gene. PPRV is closely related to rinderpest virus. Different PPRV strains are grouped
into four groups, of which 3 groups are found in Africa and 1 at Asia. One African group
is also prevalent in Asia. The Arasur strain of PPRV – AR 87, CBE (Coimbatore) and
Poondi92 are south Indian PPRV isolates.
Resistance
PPRV is stable at a pH 4.0 to 10.0. It is a heat labile virus and destroyed readily at
560C. PPRV is highly fragile virus. The virus is easily inactivated by alcohol, ether and
common disinfectants like phenol, cresol, sodium hydroxide. In chilled and frozen tissues
the virus survives for a long time.

Cultivation

Cell culture systems are ideal for virus cultivation. PPRV grows well in vero and lamb
kidney cells. The CPE develops within 5 days and consists of cell rounding, aggregation
and syncyita formation in lamb kidney cells. In Vero cells syncytia are rare and if present
are vey small. Intracytoplasmic inclusions and vacuolaion of cells are also seen.

Transmission
Affected animal excrete the virus in all secretions and excretions. Transmission
requires close contact between animals. Infection occurs mainly through inhalation of
aerosols and by ingestion. There is no known carrier state. The spread is not dependant
on vectors.
Preisposing factors for PPR infections are
1). History of recent movement or gathering together of sheep and/or goats of different
ages.
2). Introduction of recently purchased animals
3). Change in weather such as the onset of the rainy season or dry, cold periods.
4). Contact with trade or nomadic animals

Pathogenesis
PPR virus penetrates the retropharyngeal mucosa, multiplies in near by
lymphnodes sets up a viraemia and specifically damages the alimentary, respiratory and
lymphoid systems. Infected cells undergo necrosis, and in the respiratory system, also
proliferation.

Clinical features

The disease can be acute or subacute. The acute form is seen mainly in goats and
is similar to rinderpest in cattle except that severe respiratory distress is not an
uncommon feature of PPR. Acute form is characterized by high fever, dullness, sneezing
and serous discharge from the eyes and nostrils. After 2 days, discrete necrotic lesions
develop in the mouth and extend over the entire oral mucousa, forming diphtheric
plaques. Profound halitosis, sore mouth and swollen lips. Nasal and ocular discharges
become mucopurulent and the exudate dries up, mating the eyelids and partially
occuluding the external nares. Diarrhoea develops 3-4 days after the onset of fever. It is
profuse and faeces may be mucoid and blood tinged. Erosions have been described in the
vulva and prepuce. Death usually occurs within 1 week of the onset of illness. The
moratlity is 50-90% in goats and less than 10% in sheep.

Subacute forms are more common in sheep. The signs and lesions are less
marked and a few animals may die within 2 weeks, but most recover.

Necropsy findings
The carcass is severely dehydrated, the hindquarters are soiled with fluid faeces,
and crusts of exudates are present around eyes, nose and lips. Discrete or extensive areas
of erosion, necrosis and ulceration are present in the oral mucousa, pharynx, esophagus,
abomasums and distal small intestine. Haemorrhagic ulceration is marked in the
ileocaecal region, colon and rectum where they produce typical ‘Zebra stripes’. Spleen
and regional lymphnodes are enlarged. Mucopurulent exudates extend from the nasal
opening to the larynx whereas the trachea and bronchi may be hyperemic and contain
froth due to pulmonary congestion and edema. With bacterial complications, there will be
purulent or fibrinous bronchopneumonia and pleuritis.

Diagnosis
Samples to be collected
Live animals
Tears, whole blood (buffy coat), body secretions, faeces and gum debris.

Dead animals
Oral mucousa, tonsil, lungs, small and large intestines and mesenteric lymphnodes.

Diagnosis is based on
i. Classical clinical signs
ii. Virus isolation and identification in cell cultures
iii). Demonstration of viral antigen in buffy coat, body secretions, feces, lymphnode and
tonsils by immnohistochemical methods, dot-ELISA, AGID and CIEP.
Note: unlike rinderpest, PPR viral antigen is still high in tissues of animals dying from
the disease.
iv). By serology
VNT, AGID, CFT, and Complement Fixation Test are most commonly used.
More recently cELISAs have been developed based on monoclonal antibodies
specific for the N or H proteins of PPR and rinderpest viruses, and which enable
differential diagnosis of the two viruses.

Differential diagnosis

Rinderpest : In rinderpest both cattle and small ruminants are involved. The symptoms
are very severe in cattle than small ruminants.
Pasteurellosis: The disease is characterized by obvious respiratory signs, infrequent
diarrhoea, and a mortality rate rarely exceeding 10 per cent. The bipolar
organisms can be readily demonstrated in smears.
CCPP: There is no digestive system involvement, and the clinical signs and lesions are
confined to the respiratory system and pericardium.
Bluetongue: Sheep are mostly affected. Sweeling of the lips, muzzle, oral mucousa
and coronitis are more common
Contagious ecthyma / orf: Proliferative necrotic lesions are seen in the lips rather than
the whol oral cavity. The absence of nasal discharges and diarrhoea also
distinguish orf from PPR.
Prevention and control
There is no treatment for PPR. Oxytetracycline and chlortetracycline are
recommended to prevent secondary pulmonary infections. Hyper immune serum which
may be obtained from cattle hyper immunized against rinderpest can be used as therapy.
Vaccines
1. The tissue culture rinderpest vaccine at a dose of 10 2.5 TCID50 protects goats for at
least 12 months against PPR. The vaccine is currently used in many African countries
for vaccination against PPR. This vaccine interfers with rinderpest eradication
 programmes because it is impossible to determine if seropositive small ruminants
have been vaccinated or naturally infected with RPV.

2. A homologus PPR tissue culture vaccine produced by attenuation in vero cells is

commercially available. In southern India, a homologus PPR vaccine using AR-87
strain is used to control PPR in sheep and goat. This vaccine was developed at the
Department of Veterinary Microbiology, Madras Veterinary College.

3. Newly developed recombinant vaccinia or capripox viruses expressing the fusion (F)
and haemagglutinin (H) protein genes of the rinderpest virus are also effective
against PPR.

Respirovirus
Bovine Parainfluenza-3
Cause
Bovine parainfluenza virus 3 (BPI-3).
Occurrence
Bovine parainfluenza 3 virus infections occur in cattle and sheep worldwide.
Transmission
Droplet infection, direct and indirect contact.
Clinical & Pathologic Features
The virus replicates in macrophages and alveolar epithelium; the mucociliary mechanism
is adversely affected. Damage to alveolar macrophages lowers defense against bacteria.
BPI -3 virus is widely prevalent and most cattle have antibodies as a result of frequent
exposure. Seroprevalence up to 90 - 95% is common in beef and dairy cattle. Initial
exposure generally results in subclinical or mild respiratory infection. Environmental
stresses, including those incidental to transportation with over-crowding and exposure to
extremes of temperature, may lead to secondary bacterial infections with resulting
pneumonia. The most important complicating bacteria are Mannheimia (formerly
Pasteurella) haemolytica and Pasteurella multocida. This complex of a predisposing viral
infection or other primary agent with subsequent bacterial infection is often referred to as
"shipping fever" or bovine pneumonic pasteurellosis.
With secondary complications, a severe lobular pneumonia may develop with
characteristic clinical signs. If untreated this complication is often fatal.
Diagnosis
 Clinical specimens: Nasal and tracheal swabs, transtracheal wash and lung; acute
and convalescent sera.
 The virus can be isolated in cell cultures of bovine origin in which it produces
cytopathic effects characterized by giant cell formation, cell rounding, syncytia
and the production of both cytoplasmic and intranuclear inclusions.
 Examination of cryostat sections of lung tissue by immunofluorescence provides
for a rapid diagnosis of BPI-3 infection in dead animals. This can also be done in
slides prepared with cells obtained by scraping the nasal mucosa in live animals.
 A fourfold increase in antibody titer (virus neutralization, hemagglutination
inhibition, ELISA, or indirect immunoflorescence) indicates that an infection has
 been sustained. Many cattle possess antibodies to BPI-3 virus; thus, paired sera
are important.
Prevention
 Killed and live attenuated BPI-3 virus vaccines of cell culture origin are available,
usually in combination with other viral and bacterial antigens.
 Efforts should be made to minimize stress associated with marketing by allowing
calves to adjust to weaning and dehorning before they are subjected to the rigors
of sale barns, stockyards, transportation and feedlots.
 Antimicrobial drugs are used to control secondary bacterial infections.

Henipavirus
Hendra Virus Infection
(Acute equine respiratory syndrome, equine morbillivirus pneumonia)
Cause
Hendra virus, closely related to the Nipah virus.
Occurrence
The disease occurs infrequently and has only been reported in horses and humans in
Australia.
Transmission
The reservoir of the virus is fruit bats (Pteropus spp.), which occur in Australia and Papua
New Guinea. The infection in bats is subclinical. The virus is present in secretions and
urine and spread is presumed to be by direct or indirect contact and aerosol. Vertical
transmission is also thought to occur. The disease is not highly contagious.
Clinical & Pathologic Features
The principal feature of the disease is an interstitial pneumonia varying in severity.
Clinical signs are mainly those of a respiratory infection and they include fever, anorexia,
respiratory distress and a characteristic frothy, sometimes blood-tinged, nasal discharge.
Wide spread edema and neurologic signs may also be evident. The fatality rate may
exceed 60%. The virus mainly targets the vascular system, which begins in the lungs then
spreads by viral-infected macrophages to other organs and in some animals to the brain.
The virus attacks the vasculature of various tissues and organs including the spleen, liver,
kidneys, myocardium and brain.
Diagnosis
 Clinical specimens: Paired sera; lungs, spleen, liver, lymph nodes and brain.
 The virus can be isolated in a number of cell lines and identified by virus
neutralization.
 The virus can be detected in tissues by PCR.
 Testing acute and convalescent sera by ELISA or virus neutralization.
 Histopathologic examination and staining of tissues with labeled Hendra virus
antiserum.
Prevention
 The disease has been controlled by quarantine and slaughter of all infected
animals.
 Given the omnipresence of the virus reservoir there is little that can be done to
prevent infections in horses.
Public Health Significance
 The disease in humans, characterized by nonsuppurative encephalitis or interstitial
pneumonia, has been frequently fatal.
 Special precautions should be taken to prevent exposure to potential sources of
the virus.
Nipah Virus Infection
(Barking pig syndrome, porcine respiratory and neurologic syndrome)
Cause
A recently discovered paramyxovirus referred to as Nipah virus, which is closely related
to Hendra virus.
Occurrence
The virus was first recovered from humans with encephalitis in Malaysia and Singapore
in 1998 - 1999. It was determined that those infected had been exposed to infected pigs.
There is evidence that in addition to swine and humans, horses, dogs and cats are
susceptible. The reservoir of the virus is considered to be fruit bats of the genus Pteropus
that occur widely in southeast and south Asia.
The outbreaks in Malaysia and Singapore were controlled and there have been no further
outbreaks in swine.
Transmission
After introduction of the virus to swine herds spread is rapid and presumably by direct
and indirect contact. Further evidence of the high infectivity of the virus was rapid spread
among herds.
Clinical Features
A febrile respiratory infection with rapid, labored breathing develops in many pigs in a
herd. This gives rise to a severe characteristic cough and thus the name barking pig
syndrome. Less common than the respiratory form was an encephalitis seen mainly in
boars and sows. The mortality rate in pigs (> 5%) was lower than that in humans.
Diagnosis
 Isolation of the virus in cell cultures and identification of the virus by serologic
means, including virus neutralization and immunofluorescence. Syncytia are
produced in Vero cells.
 Viral antigens in tissues can be identified by chemically tagged specific
antibodies.
Prevention
 The disease was controlled in Malaysia and Singapore by strict quarantine of
premises and slaughter of all the swine in affected herds.
 Because of the bat reservoir of the virus constant surveillance for evidence of the
disease must be exercised.
Rubulavirus
Porcine Rubulavirus Infection
(Swine blue eye disease)
Cause
Porcine rubulavirus.
Occurrence
Porcine rubulavirus infection (PRI) of swine was first reported in Mexico in 1980 and
still occurs there. It has not been reported as occurring outside of Mexico.
Transmission
The disease is acquired by direct and indirect contact (fomites).
Clinical & Pathologic Features
PRI or swine "blue eye" disease is most severe in pigs younger than three weeks and is
characterized clinically by sudden onset of fever, depression, and progressive CNS signs.
Affected pigs are weak and ataxic, and may have rigidity of the hind legs and tremors.
Dilated pupils, nystagmus, and conjunctivitis may be noted in some pigs, and
approximately 1 - 10% of affected pigs develop corneal opacity. The mortality rate may
approach 90%. In older pigs, clinical signs are principally respiratory including sneezing
and coughing, and anorexia and fever. Neurologic signs may be seen in pigs more than
30 days old. The mortality rate is usually low. Most infections in adult swine are
subclinical; corneal opacity is occasionally observed. Reproductive problems may occur
in pregnant sows. Orchitis and epididymitis may occur in boars resulting in reproductive
failure.
Gross necropsy lesions are minimal and nonspecific. Microscopic lesions are those of a
nonsuppurative encephalomyelitis and interstitial pneumonitis.
Diagnosis
 Clinical specimens: Brain, lungs, tonsils and affected eyes.
 A presumptive diagnosis has been made on the basis of clinical signs and
histopathologic lesions. Epidemiological data is supportive.
 Confirmation requires isolation and identification of the virus. The virus can be
cultivated in cell cultures of swine origin (PK-15 cell line) in which it produces
syncytia. It is identified by virus neutralization.
 The virus agglutinates erythrocytes of various animal species including the
chicken.
 Serological testing of paired sera using hemagglutination-inhibition and ELISA.
Prevention
 This is a reportable disease.
 Prevention is best accomplished by maintaining closed herds. All replacement
animals should be isolated and serologically tested.
 Inactivated vaccines have been used.

Canine Parainfluenza Virus 2

This rubulavirus causes subclinical to usually mild respiratory infections in dogs.

It may be part of the etiology of kennel cough, together with other microbial agents,
including Bordetella bronchiseptica, canine adenovirus, canine herpesvirus, reovirus,
mycoplasmas and possibly others.
Diagnosis of kennel cough is based on history and clinical signs. Canine
parainfluenza virus 2 may be included in a multiple component vaccine given to dogs
prior to exposure in boarding kennels, shows, etc.

Pneumovirus
Bovine Respiratory Syncytial Virus Infection
Cause
Bovine respiratory syncytial virus. (BRSV)
Occurrence
Affects cattle, sheep and goats and is widely distributed in cattle throughout the world.
The disease occurs mainly in young beef and dairy cattle.
Recently weaned and transported calves are particularly susceptible.
Transmission
Infected cattle shed virus in respiratory secretions, and the disease is spread by
respiratory droplets and by direct and indirect contact.
Clinical & Pathologic Features
The virus replicates in the respiratory epithelium and cause necrosis. Syncytia and
intracytoplasmic inclusion bodies are seen in bronchiolar, alveolar epithelial and other
cells.The morbidity is generally high in fully susceptible herds; the mortality may
approach 20% in severe outbreaks.
The occurrence of moderate to high levels of BRSV antibodies in herds not experiencing
respiratory disease in the recent past suggest that many BRSV infections may be
subclinical or mild. However, some animals develop acute diffuse interstitial pneumonia
with or without secondary bacteria. Mortality is not rare in bacteria-complicated, severe
cases. Clinical signs include coughing, nasal discharge, and fever. Severely affected
animals may display dyspnea and mouth breathing. In the absence of secondary bacterial
infections, recovery occurs in 1 - 2 weeks.
Diagnosis
 Clinical specimens: Slide preparations from nasal and conjunctival scrapings,
nasal and conjunctival swabs, lung, and acute and convalescent sera.
 A rapid diagnosis can be achieved by the fluorescent antibody (FA) examination
of cytologic preparations of nasal and conjunctival epithelia collected early in the
course of disease.
 Similarly, FA or immunoperoxidase staining is used to demonstrate viral infected
cells in cryostat sections of lung tissue in animals that have died. In these animals
histopathologic lesions are helpful in making a diagnosis of BRSV infection,
especially if syncytial cells containing cytoplasmic inclusions are observed.
 The virus, which is antigenically related to human respiratory syncytial virus, can
be isolated in cell cultures of bovine origin in which it produces cytopathic effects
consisting of syncytia and eosinophilic cytoplasmic inclusions.
 Because of the lability of the virus, isolation attempts are apt to be negative unless
the specimens are almost immediately fresh.
 A serologic diagnosis can be made by the demonstration of a significant increase
in antibody levels between acute and convalescent sera using ELISA or virus
neutralization.
Prevention
 Important considerations are: a closed herd policy, raising young animals apart
from older animals, strict sanitation measures and vaccination.
 Modified live and inactivated virus are included in combined products to prevent
other important diseases such as infectious bovine rhinotracheitis, bovine virus
diarrhea and bovine parainfluenza 3 infection.

 Metapneumovirus
Turkey Rhinotracheitis
This disease is caused by the only species in the genus Metapneumovirus. Three and
possibly more types of the virus have been identified to date.
The disease affects turkeys (3 - 10 weeks) and probably occurs worldwide. It spreads
rapidly by infectious droplets.
It is a severe, highly contagious upper respiratory infection characterized by sudden
onset, swollen sinuses (swollen head syndrome) and coryza. Morbidity and mortality may
be high.
Because of its similarity to other respiratory diseases, diagnosis should be confirmed by
laboratory identification of the virus. Procedures used are isolation and identification of
the virus using turkey organ cultures or embryonated eggs. Various serological
procedures are available for flock surveys; ELISA is probably the most satisfactory.
Live attenuated and oil adjuvant vaccines are used for prevention.

RHABDOVIRIDAE
Based on virion properties and serologic relationships, four genera containing
animal viruses have been recognized in the family Rhabdoviridae, the genera Lyssavirus,
Vesiculovirus, Ephemerovirus and Novirhabdovirus. Lyssavirus, Vesiculovirus and
Ephemerovirus genera contain viruses, which infect vertebrates. Infectious
haemotopoietic necrosis virus and related rhabdoviruses of fish are included in the genus
Novirhabdovirus.

Order: Mononegavirales
Family: Rhabdoviridae

Genus: Lyssa virus

Species: Rabies virus,
Rabies related viruses (Lagos bat virus, Australian bat virus)

Genus: vesiculovirus
Species: Vesicular stomatits – Indiana
Vesicular stomatits – New Jersey virus.

Genus: Ephemero virus

Species: Bovine ephemeral fever virus

Genus: Novirhabdovirus
Species: Infectious haemotopoietic necrosis virus
Rhabdoviruses of fish.

Virion properties
Rhabdovirus virions are 70 nm in diameter and 170 nm long and consists of an
envelope with large peplomers within which is a helically coiled cylindrical nucleocapsid
which gives a distinct bullet or conical shape.
 The genome is a single molecule of linear, negative sense, single stranded RNA,
11 to 15 kb in size. The genome encodes five genes in the order 3’-N-NS-M-G-L-5’. The
viruses generally have five proteins:
L - the RNA-dependent RNA polymerase that functions in transcription and RNA
replication.
G - the glycoprotein contains neutralizing epitopes, which are targets of vaccine-induced
immunity.
N - the nucleoproteins, the major component of the viral nucleocapsid
NS - a component of the viral polymerase
M - a protein that facilitates virion budding by binding to the nucleocapsid and to the
cytoplasmic domain of the glycoprotein.
Three proteins (N, NS and L), in association with viral RNA, constitute the
nucleocapsid. The virus replicates in the cytoplasm. Maturation is by budding through the
plasma membrane. Vesicular stomatits viruses cause rapid cytopahology where as street
rabies virus are non cytopathogenic.
Viral Replication
Viral entry into its host cell occurs via fusion of the viral envelope with the cell
membrane; all replication steps occur in the cytoplasm. Replication first involves mRNA
transcription from the genomic RNA via the virion polymerase; later, using the protein
products of this transcription, there is production of full length, positive stranded
templates, which in turn are used for the synthesis of genomic RNA. Using virion RNA
as template, the viral transcriptase transcribes five subgenomic mRNA species. There is
only a single promoter site, located at the 3’ end of the viral genome; the polymerase
attaches to the genomic RNA template at this site and it moves along the viral RNA, it
encounter stop/start signals at the boundaries of each of the viral genes. Only a proportion
of polymerase molecules move past each junction and continue the transcription process.
This mechanism, called attenuated transcription or stop/start transcription or stuttering
transcription

RABIES

Synonym: Hydrophobia, Lyssa, Mad dog disease

Rabies is acute viral encephalitis of all warm-blooded animals characterized by

altered behaviour, aggressiveness, progressive paralysis and in most species by death. It
is an important disease of man and animals, which is usually transmitted through the bite
of rabies animals to other animals or human result in rapid fatal encephalomyelitis after a
somewhat lengthy incubation period.

Note: The disease in man is called hydrophobia because the patient exhibits fear of
water, beig incapable of drinking though subject to intolerable thirst. Rabies in animals is
not called hydrophobia because they do not have this peculiar feature.

Host
 Man and warm-blooded animals. Fox, wolf and jackal are extremely susceptible.
Guineapig, Hamster, Bat, Mongoose, Mice, Rabbit, Skunk and cattle are highly
susceptible. Dog, sheep, goat, horse and human are moderately susceptible. Poultry and
opossum are resistant.

History

Rabies is one of the oldest and most feared diseases known to mankind. The
disease was recognized in Egypt before 2300 B.C. The name rabies comes from the latin
word rabidus, meaning mad, derived from the Sanskrit root rabhas, to do violence. In
1804 Zinke first demonstrated the infectious nature of saliva from a rabid dog. In 1826
the ‘poison’ was described as going along nerves into the CNS, an observation that still
holds true. Pasteur and his collegues in 1881-1889 discovered that the disease was caused
by an infectious agent and that the CNS was the actual site of infection. In 1885 Pasteur
prepared the first human inoculation with antirabies vaccine on 9-year-old boy (Joseph
Meister) who had been bitten 14 times by a rabid dog. He was inoculated with rabies
infected spinal cord tissue that had been preserved in a flask for 15 days at room
temperature. He did not develop rabies after being given 13 injections of the spinal cord
material.

Distribution

At the present time rabies occurs in most parts of the world except in Japan, UK,
New Zealand, Antarctica, Australia, Hawaii islands and Switzerland. In India the
incidence is very high.

Properties
Rabies virus belongs to the Rhabdo virus group, one of the RNA helical
enveloped virus group. It appears as filamentous, bell shaped or bullet shaped and
approximately 60  175 nm in size. The virion is rounded at one end and flat or
truncated at the other. It is surrounded by a fringe of short pointed projections. Rabies
virus can agglutinate RBC’s of day old chicks and goose at 0-4 0C and pH 6.2. The virus
is deficient in neuraminidase activity. The nucleic acid consists of single stranded RNA.
It is found in nerve tissue, saliva, salivary glands, pancreas, less often in Urine and lymph
and rarely in blood, milk and other body fluids of infected animals.

It is readily inactivated by sunlight, drying in air, 40-70% alcohol, Quarternery

ammonium compounds, tincture-iodine, carboilic acid and any lipid solvents. The virus is
very resistant to autolysis and putrefaction. The virus will persist for months in infected
nervous tissue in 50% glycerol. The virus survives at 4 0C for weeks. It can be preserved
at –700C or by lyophilization.

Antigenic property
The surface spikes are composed of glycoprotein, which induces the formation of
glycoproteins. (The purified glycoprotein may be used effectively as subunit vaccine)
 All strains of unmodified rabies virus isolated from natural human or animal
infection is termed street virus resembling each other closely in antigenic structure. Fixed
virus is produced from street virus by repeated serial passages to other animals. The
characteristic of fixed virus is

i) Progressive shortening of length of incubation period, which remains

constant and irreversible.
ii) Failure to produce Negri bodies.
iii) Increased virulence for its natural host
iv) Decreased virulence for other species.
v) Used for vaccine production.

The fixed strains that have been adapted in egg are Flurry strain and Kelev strain.
Low egg passage (LEP) strains can be obtained by 40 to 50 passages. High egg passage
(HEP) strains can be obtained by 180 passages.

Cultivation of virus

Cell culture
Rabies virus grows very well in human diploid cell line WI-38 and MRC-5,
BHK21, Vero and Chicken embryo fibroblast. Cytopathic effects are not apparent. In
continuous lines of human diploid cells and BHK21 infected with rabies, almost 100% of
the cells are infected and there is a marked CPE
Embryonated eggs
Five or six day old embryonated hen’s eggs are preferred for growth of virus by
yolk sac route. The vrus also grow in CAM route and produce miliary pock lesions.
The strains that have been adapted in egg are Flurry strain, Keller strain. Low egg
passage (LEP) strains can be obtained by 40 to 50 passages. High egg passage (HEP)
strains can be obtained by 180 passages.

Duck eggs are commonly used for preparation of vaccine.

Labortory animals

Rabies can be cultivated in a wide range of laboratory animals such as hamsters,

mice, guinea pig and rabbits in the order of decreasing susceptibility by intracerebral or
intramuscular routes.

Epidemiology

In usual circumstances the only risk of rabies virus transmission is by the bite or
scratch of a rabid animal, although in bat caves, where the amount of virus may be very
high and the extremely high humidity may stabiize the virus transmitted by aerosol
infection. In rare circumstances the virus can pass through the intact mucous membrane
 The amount of virus required and to initiate infection varies considerably and
depends upon the susceptibility of the patients. Susceptible wild living animals
particularly foxes, wolf, jackals constitute the most important residual focus of rabies
infection the most prevalent and most difficult to control. Because they carry 10 6
infectious units of virus per mili liter of saliva.

Domestic dog is still the most important source of infection. Although virus titres
in the saliva of confirmed case of rabies in dog varies widely the saliva is believed to be
infective in only 50-60% of the rabid dogs at the time of bite and the presence of virus is
greatly evident in a few days before death. The factors, which influence the course of
infection, include the virulence, invasiveness and concentration of inoculated virus, the
amount of nervous tissue near the site of wound, the degree of trauma and the type of
animal inflicting the bite. Blood licking bats in chiroptera family constitute an important
reservoir in central and South Africa and they frequently infect cattle, man by biting at
nights. In a report from Ethiopia in 1964, the virus isolation of true rabies virus was
obtained from the saliva of dog, which remains clinically healthy 4 years after the sample
had been taken.

Pathogenesis
Following introduction into the tissues, virus enters peripheral nerve endings.
There may be limited replication locally in myocytes or other tissue cells and from there
begins to travel centrifugally along the nervous routes by axoplasmic flow. The virus may
spread from the brain along peripheral nerves to reach other tissues particularly in
salivary glands where it multiplies and enters the saliva, which is the commonest source
of infection in bite wound.

Depends upon the location and severity of the bite the incubation period in natural
outbreak of dog rabies averages from 3-8 weeks. It may extends upto one year.

Bite delivered virus deep in to striated muscle and connective tissue

Virus gets amplified by replicate in myocytes (muscle cells)

Binds with acetyl choline ( Neurotransmitter at neuromuscular junction)

Which facilitates entry into nerves

Invades peripheral nervous system

Centripetal passive movement within axoplasm

 Deliver virus into the CNS, usually via the spinal cord

Reaches the limbic system of the brain & replicates

extensively (Furious form)

Spread within CNS (replicate in Neocortex) – dumb form

Moves centrifugally from CNS through peripheral nerves

Reaches various organs including salivary glands

Virions released by budding through plasma membrane

High concentration of virus in saliva

Although rabies proteins are highly immunogenic, neither humoral or cell

mediated response, can be detected during the stage of movement from the site of bite to
CNS. Because very little antigen is delivered into the system.

Clinical signs
The incubation period in natural outbreak of dog rabies averages from 3-8 weeks.
But it may be as short as 10 days to as long as one year. Dog has a change in the
temperament aimless snapping and barking at imaginary objects. By about 3 rd day after
the onset of illness the dog enters the furious stage which lasts for 3-7 days. During this
stage dog becomes irritable, restless, nervous, deprived appetite, characteristic change in
its barking and may howl in an unusual tone due to the paralysis of laryngeal muscle.
Salivation and frothing at the mouth becomes progressively more profound. Convulsions
and complete muscular co-ordination are present shortly before the animal dies. If it does
not die it passes into paralytic stage.
In cases when the furious phase is extremely short or absent the animal rapidly
enters the paralytic or dump stage; where the dog is rarely irritable and seldom bites
lower jaw paralysis, and complete in coordination of movement and death.

The clinical features divided into three phases.

Prodromal form: During the prodromal period, which lasts 1-3 days, animals
show only vague CNS signs, which intensify rapidly. Dog has a change in the
temperament aimless snapping and barking at imaginary objects.
 Furious form: By about 3rd day after the onset of illness the dog enters the furious
stage which lasts for 3-7 days. During this stage dog becomes irritable, restless, nervous,
deprived appetite, aggressive, and often dangerousas it loses all fear of humans and bites
at anything that gains its attention. Exaggerated response to light and sound. Dog exhibit
characteristic change in its barking and may howl in an unusual tone due to the paralysis
of laryngeal muscle. Salivation and frothing at the mouth becomes progressively more
profound. Termianlly, there are often convulsive seizures, coma and respiratory arrest,
with death occurring 2 to 14 days after the onset of clinical signs. If it does not die it
passes into paralytic stage.

Dumb or paralytic form: In cases when the furious phase is extremely short or
absent the animal rapidly enters the paralytic or dump stage; where the dog is rarely
irritable and seldom bites. Paralysis of the throat and masseter muscles. Droppings of the
lower jaw or lower jaw paralysis are common in dogs. In dump form animals are not
vicious and rarely attempt to bite. The paralysis progresses rapidly to all parts of the
body, and coma and death follow in a few hours.

Note: Veterinarians / Owners frequently examine the mouth of dogs and livestock
searching for foreign body or admiister medication with their bare hands, therby exposing
themselves to rabies.
In cats the clinical signs are similar to those of dogs but lost for 2-4 days before
death occurs.

Cattle: Among farm animals, cattle are most commonly affected. In the parlytic form,
knuckling of the hind fetlocks, sagging and swaying of the hindquarters while walking,
often deviation or flaccidity of the tail to one side, are common signs. Decreased
sensation over the hindquarters is one of the best criterions for the detection of rabies.
Drooling of saliva, tenesmus, pumping of anus and followed by recumbency seen in later
stages. In furious rabies, the animal alert, hyper sensitive, violently attack, loud and
coarse bellowing, sexual excitement and collapses suddenly

Sheep: Clinically the picture is similar to cattle. Sexual excitement, violent attack,
vigorous wool pulling, sudden falling and salivation are characteristic. Goats are
commonly aggressive, and continuous bleating is common.

Horse: Muzzle tremors and pharyngeal pareis are common. In addition to this abnormal
postures, frequent whinnying, kicking, biting, colic, sudden onset of lameness in one limb
followed by recumbency, high stepping gait, blindness, recumbency, paddlig,
convulsions and terminally paralysis.
Pigs: Tendency to attck, twitching of the nose, rapid chewing movements, excessive
salivation, walk backward and terminally paralysis.
Cattle are very restless, excited and aggressive with salivation, abdominal pain,
diarrhoea and rectal straining. Paralysis of hind quarters occur followed by death in 3-6
days after the first signs of illness. In sheep and goat the symptoms are similar to cattle.

Diagnosis
Samples to be collected
Live animals: Saliva, corneal/ Conjuctival smear

Dead animals: The whole carcass or the severed head of the animal suspected to have
died of rabies. Alternatively, the brain may be removed carefully and two portions, one in
50% glycerol saline and the other in Zenker’s fixative, sent for biological test and
microscopy, respectively.

The brain tissue selected should include portions of hippocampus, cerebellum,

cerebral cortex, and placed in 50% glycerol saline to preserve the virus. No refrigeration
is required. Glycerinated tissues are generally unsuitable for immunofluorescence
staining for which smears fixed in acetone free methyl alcohol and dried at room
temperature without blotting should be sent.

Demonstration of Negribodies: Negribodies are abundantly present in pyramidal cells

of ammon’s horn and purkinje cells of cerebellum. They can be demonstrated in
impression smears either by Seller’s method (for unfixed sections) or Mann’s method
(fixed sections). Seller’s method has the advantage that fixation and staining is done
simultaneously. Negribodies are intracytoplasmic, magenta red or cherry red spherical or
oval bodies with characteristic basophilic inner granules. They are varying in size from 3-
27µ. They are inclusions of aggregates of nucleocapsids and they are specific to rabies
infection.

Note: Animal should not be killed for the demonstration of negri bodies. They can be
demonstrated in only 80-85% cases. It should be differentiated from intranuclear
inclusions produced by Infectous Canine Hepatitis and Canine Distemper.

Isolation and identification of virus

Confirmatory diagnosis must be obtained by animal inoculation or other means

Animal inoculation: For isolation of virus the brain or other tissue specimens are
prepared as 10% suspension. Suckling mice or hamsters of 3-6 weeks age are generally
used for isolation and identification. Route of inoculation is intracerebral. Infected mice
almost always develop clinical symptoms with in 17 days of inoculation. The clinical
signs to look far in mice are ruffled fur, arching of back, flaccid paralysis of legs. To
establish a positive diagnosis the mouse brain are removed and examined microscopically
for negribodies. The confirmatory diagnosis can be obtained by serum neutralization test
in mice or by lmmunofluorescence.

Rabies virus can be cultured in human diploid cells, neuroblastoma and BHK21.
5 or 6 day old embryonated hen’s eggs are preferred for growth of virus by yolk sac
route.

Demonstration of viral antigen

Demonstration of viral antigen in ammon’s horn, cerebellum, corneal/conjunctival
impression smear, skin biopsy ( from facial area ) and in submaxillary salivary glands by
fluorescent antibody technique (FAT) is the most widely used, confirmatory test for the
diagnosis of rabies since it provides rapid reliable and highly specific means of detecting
the viral antigen. This test is recommended by both WHO and OIE.

By serology
Immunoperoxidase test. ELISA, Rapid fluorescence focus inhibition test
(RFFIT), Virus neutralization test, Rapid rabies enzyme immuno diagnosis test (RREID)
based on the use of antinucleocapsid lgG are highly useful.
RT-PCR amplification technique is 1000 times more sensitive than other tests.

Treatment
There is no specific treatment for rabies. Dogs usually die after showing clinical
signs. The site of bite should be washed with water or soap. Alkali prevents
multiplication of virus. Sodium bicarbonate or caustic soda may be used. Then the wound
is treated with 2% quarternary ammonium compound or tincture iodine or 40-70%
alcohol. Wound may also be cauterized with carbolic acid or nitric acid. Antirabic serum
may be applied topically or infiltrated around the wound. Treatment with antirabic serum
is also effective.

Antirabic serum is prepared by hyperimmunisation of horses. It should be given

as early as possible after exposure, and in any case within five days, after which it may
not be beneficial. The dose recommended is 40 I.U. per kg body weight.

Prophylaxis
Nervous tissue vaccine
This is a flury type vaccine. These are prepared from brain tissues of sheep
inactivated through 1% phenol. These are made as 5% and 20% suspension.

Pasteur’s cord vaccine: this vaccine is prepared by drying over caustic potash for
varying period’s pieces of infected rabbit spinal cord.

Semple vaccine: This vaccine was developed by Semple (1911) at the Central Research
Institute, Kasauli. It is a 5% or 20% suspension of sheep brain infected with fixed virus
and inactivated with phenol at 370C.
Beta Propiolactone vaccine: This is a modification of Semple vaccine in which BPL is
used as the inactivating agent instead of phenol.

UV treated vaccines (Webster) : Inactivated vaccines prepared from brain of infected

animals, which were exposed to UV rays.

Non- nervous tissue vaccine

Egg vaccine

Duck egg vaccine : This is fixed virus adapted for growth in duck eggs and inactivated
with BPL. This has been discontinued because of its poor immunogenicity.

Live attenuated chick embryo vaccine: Two types of vaccines are available.

Avianized or chick embryo vaccine

This is made through passaging the virus in chick embryo. Low egg passage (LEP) (40 to
50 passages) is completely safe for dogs. Avianized live virus vaccine is a single dose
vaccine. Age of first vaccination is 3 months, dose 3 ml i/m. Immunity 3 years. Repeat
after 3 years.

High Egg Passage (HEP) vaccine at 180 passage level for cattle and cats.
Tissue culture vaccine
The first cell culture vaccine was the human diploid cell strain (HDCS) vaccine
developed by Koprowsky, Wiktor and Plotkin. It is a purified and concentrated
preparation of fixed rabies virus (Pitman – Moore Strain) grow on human diploid cells
( WI 38 or MRC 5) and inactivated with betapropiolactone. It is highly antigenic and free
from side effects. Its only disadvantage is high cost.
Nowadays more economical, primary chiken embryo and vero cell culture
adapted, vaccines are available.

The avianized live virus vaccine is a single dose vaccine. Age of first vaccination
is 3 months, dose 3 ml i/m. Immunity 3 years. Repeat after 3 years. In tissue culture
vaccine, age of first vaccination is 3 months. Revaccination after 6 months then at yearly
intervals. Dose 5 ml s/c in the neck or flank region.

Subunit vaccine

The glycoprotein subunit on the virus surface, which is the protective antigen has
been cloned and recombinant vaccines produced. They are still in the experimental stage.

Post exposure immunization

If the dog is not protected earlier: Dog weighing less than 15 kg should get 2 ml
s/c for 14 days. Dog weighing 15 kg or more should receive 5 ml s/c for 14 days.
If the dog is protected earlier: Dog weighing less than 15 kg should receive 2 ml
s/c for 7 days. Dog weighing 15 kg or more should receive 5 ml s/c for 7 days.
Control (as per the WHO recommendations)
1. Notification of suspected cases, and destruction of dogs with clinical signs and dogs
bitten by a suspected rabid animal.
2. Compulsory immunization of dogs
3. Sterlization and Vaccination of stray dogs by using baits
4. Epidemiological Surveillance
5. Education of Public
6. Development of cost effective vaccine

VESICULAR STOMATITIS

This febrile disease affects mainly horses, donkeys and cattle and pigs. Other
susceptible species include camels, several wildlife species and humans. Vesicular
stomatitis is clinically similar to foot and mouth disease. Vesicular stomatiitis virus
belongs to number of closely related, antigenically distinct members of the genus
Vesiculovirus, of which the type species vesicular stomatitis Indiana virus can cause the
disease. Most outbreaks are associated with vesicular stomatitis Indiana virus or with
vesicular stomatitis New Jersey virus, a more virulent virus.

Epidemiology
The disease is limited to the western hemisphere and the infection is endemic in
Central America and in regions of South America. Outbreaks of the disease occur every
two to three years in tropical and subtropical regions, with clinical cases most common at
the end of the rainy season and early in the dry season.

Transmission
The virus can be biologically transmitted by black flies and mechanically by
culicoides, houseflies. The saliva and vesicular fluid from clinically affected animals are
highly infective but infectivity diminishes rapidly and may be lost within 1 week after the
vesilce rupture. Frrom this source, virus may be transmitted by fomites, such as
contaminetd feed, milking machine and restraint devices. Recovered cattle act as
convalascent carrier for 38 days to 5 months.

Pathogenesis
Virus probably enters the body through abrasions on the skin or mucous
membranes or following an insect bite or by rough forages. As in FMD, there is a
primary viremia with subsequent localization in the mucous membrane of the mouth and
the skin around the mouth and coronets. Absence of classical vesicles frequently
encountered. Vesicles, which develop at the site of infection, may coalesce. Spread may
occur locally by extension from primary lesions. Although secondary lesions at distant
sites may develop, it is unclear how transfer of the virus occurs and if these lesions result
from viremia or following environmental contamination.

Clinical features
The incubation period is up to five days. Subclinical infection is common.
Affected animals, which are usually more than one year old, become febrile. Vesicles
develop on the tongue and on oral mucous membranes, often accompanied by profuse
salivation. Secondary lesions may occur on the coronary band and teats. Lameness is
often a prominent feature of the disease in pigs. Mastitis may develop in cows with
severe teat lesions. In the absence of secondary infection, lesions generally heal within
two weeks.
The economic impact of the disease relates to production losses, culling and other
disease control measures. Following infection, animals develop high levels of
neutralizing antibodies but the duration of protection is variable. Cross-protection
between vesicular stomatitis Indiana virus and vesicular stomatitis New Jersey virus is
limited.

Cattle: The incubation period is 3-15 days. There is a sudden appearnce of mild fever
and the development of vesicles on the dorsum of the tongue, dental pad, lips and the
buccal mucousa. Development of vesicles accompanied by ropy salivation. Vesicles
ruptured followed by erosive necrotic lesion. Subclinical infection is common. Affected
animals, which are usually more than one year old, become febrile. Secondary lesions
may occur on the coronary band and teats. Mastitis may develop in cows with severe teat
lesions. In the absence of secondary infection, recovery is rapid, affected animals are
clinically normal in 3-10 days. Cross-protection between vesicular stomatitis Indiana
virus and vesicular stomatitis New Jersey virus is limited.

Horses: There is fever, depression and drooling of saliva. Affected horse may rupture
their lips on troughs. Vesicles coalesce, rupture and formation of shallow ulcers. Lesions
are also seen in coronary band and teats.

Pigs: Lameness is more common.

Diagnosis
Prompt laboratory confirmation is required because of similarities between
vesicular stomatitis, foot-and-mouth disease and swine vesicular disease. If horses
present with vesicular lesions, infection with vesicular stomatitis virus should be
considered.
• Suitable specimens for isolation of virus or the detection of viral antigen include
epithelium from lesions and vesicular fluid.
• Viral antigen can be detected by CFT or ELISA.
• Virus may be isolated in suitable cell lines, in embryonated eggs or in suckling
mice by intracerebral inoculation. FAT, ELlSA, CFT or the virus neutralization tests are
suitable for identification of the isolates.
• Electron microscopy can be used to identify virus in specimens or tissue culture.
• Antibody levels in recovered animals may be assayed by CFT, the virus
neutralization test, competitive ELISA or lgM-specific capture ELISA. Because levels of
complement fixing and IgM79 antibodies persist for only short periods, assays based on
procedures involving these antibodies can be used to confirm recent infections in
endemic areas.

Treatment and control

 • Specific treatment is not available. Measures aimed at minimizing secondary
infections may be beneficial.
• Movement restrictions and a 30-day quarantine period following the last
clinical cases are recommended for infected premises.
• Insect-proof buildings and avoidance of habitats associated with insect vectors
reduce the likelihood of infection.
• Although both inactivated and attenuated vaccines have been used, they are not
commercially available.

BOVINE EPHEMERAL FEVER

Synonym: 3-day sickness, 3-day fever, Dragon Boat disease

BEF is an acute, non-contagious, arthropod-borne viral disease of cattle and water

buffalo characterized by sudden onset, lameness and quick recovery with high morbidity
and low moratlity. This disease occurs in tropical and suntropical regions of Africa, Asia
and Australia.

Properties
The BEF virus belongs to the genus Rhabdovirus. Morphology, resistance and
replication are similar to other rhaboviruses. The virus has no HA property. The virus is
rapidly inactivated at pH levels below 5

Virus strains
This virus is antigenically related to at least three other viruses non pathogenic for
cattle. Kimberly virus, Berrimah virus, Adelaide river virus and two that produce an
ephemeral fever like diseseas in cattle, kotonkah and Puchong virus.

Cutlivation
Isolation of BEFV is very difficult. BEFV are cultivated through intravenous
inoculation in chiken embryos. Lesions are not characterstic and infected embryos die.
The virus can be cultivated in suckling mice, vero cells and in mosquito cells.

Transmission
The virus transmitted mainly by vectors. Mosquitoes (Aedes, Anophles and
Culex), and biting midges are playing a major role. Strong wind can transport vectors
longer distances. Cattle are the only reservoir host. Epizootics associated with recet
rainfall. The virus does not spread by any other modes.

Pathogenesis
BEFV has affinity with leucocyte –platelet fraction of the blood. Blood-sucking
insects acquire the virus when feeding on animals during the brief viraemic stage of the
disease. The virus is shed in its saliva and is transmitted to a new host through wounds
during feeding. Virus multiplication primarly occurs within the vascular system is
followed by generalized inflammation.
 Many of the changes observed in infected animals are attributed to host response
rather than to direct viral damage.

Clinical features
The incubation period is 2 to 10 days. Calves are least affected, those less than 6
months of age showing no clinical signs. After an incubation period of 2-4 days, there is
sudden onset of fever. A biphasic or triphasic fever and sharp fall in milk yield are
commonly observed. There is severe ruminal stasis followed by constipation. Muscular
signs become more evident on the second day with severe stiffness. Swellings in
shoulders, neck and back may be seen. A posture similar to that of acute laminitis, with
all four feet bunched under the body, is charcteristic. On the third day the animal begins
eating and ruminating, and the febrile reaction disappears.

Some animal’s remains standing in acute stage and exhibit sternal recumbency
associated with hypocalcemia. (Milk fever posture). Most recovered animals develop a
solid immunity. The morbidity rate may reach 100% and the fatrality rate is 1%. Milk
yield returns to normal within 2 to 3 weeks.

Lesions
Vasculitis, serofibrinous polyserositis, sinuvitis, pulmonary emphysema,
accumulation of odematous fluid in pericardium and pleura are seen. All lymphnodes are
usually enlarged. Congestion and petechiae is often seen.

Diagnosis
Blood (Buffy coat) and serum is the highly suitable specimen.
Based on clinical signs
Based on clinical pathology
Marked leukocytosis, increased plasma fibrinogen, high level of creatinine kinase,
low level of calcium are common.
Based on serology
CFT, AGID, Virus neutralization test, FAT or blocking ELISA are commonly
employed.

Treatment
Affected animals should be rested. Anti-inflammatory drugs such as
phenylbutazone proved useful for treatment. Intravenous or subcutaneous administration
of calcium borogluconate is recommended. Oral drenching should be avoided during the
acute phase of the illness because swallowing may be impaired.

Control
Vector control is usually impractical in endemic areas. Vaccination is the only
effective method of control. Both formalin killed and attenuated vaccines are available.
Trials have been carried out using a limit vaccine based on the envelope-
glycoprotein has been developed.
 Filoviridae
This family of enveloped, negative-sense single-stranded RNA viruses consists of what
are termed Marburg-like viruses and Ebola-like viruses. These viruses, which occur in
Africa, cause hemorrhagic fevers with high fatality rates in humans and some primates.
Viral Characteristics
 They are enveloped viruses with a helical nucleocapsid and linear single-stranded,
negative sense DNA.
 They are morphologically distinctive in being pleomorphic and having filaments
as long as 1400 nm; capsids are as long as 800 nm.
 The method of replication is similar to the Rhabdoviridae and Paramyxoviridae.
 There is a lipid envelope with peplomers (10 nm long).
 Virions contain seven structural proteins.
Significance
Filoviruses were first recovered from cases of hemorrhagic fever in German laboratory
workers in the late 1960’s. The Marburg virus involved was traced back to African green
monkeys imported from Uganda.
Approximately one decade later, Ebola virus was recognized as the agent of
hemorrhagic fever in outbreaks in Zaire and Sudan.Eventually a similar virus was
brought into the USA by macaque monkeys imported from the Philippines. Since then,
sporadic outbreaks of the Ebola virus-associated disease have been reported in several
African countries. In these outbreaks, introduction of the virus to human population
seems to have been via an infected non-human wild host. Ebola virus is among the most
lethal viruses for humans and is classified as a biosafety level 4 agent.
Although Ebola and other filoviruses are clearly zoonotic, the natural reservoirs for these
viruses remain unknown and thus constitute a major challenge for epidemiologists.

Bunyaviridae and Bornaviridae

Both families consist of negative-sense, single-stranded RNA viruses.
Bunyaviridae is a large family of mainly arthropod-borne viruses (arborviruses) with
several significant veterinary pathogens. Bornaviridae has only one species and horses
and sheep appear to be the principal natural hosts.
Bunyaviridae
Bunyaviridae is the largest family of vertebrate viruses. Most bunyaviruses are
transmitted by biting arthropods and, with the exceptions of Akabane disease, Cache
valley, Rift valley fever, and Nairobi sheep disease, are of limited veterinary importance.
Viral Characteristics
 These viruses are 80 - 120 nm in diameter, have a helical nucleocapsid
surrounded by an envelope, on the surface of which are glycoprotein projections.
 The genome consists of three segments of negative-sense single-stranded RNA.
 The segmented RNA genome may undergo genetic reassortment leading to new
strains.
 With most of these viruses, genes are expressed in two disparate host systems,
vertebrate and arthropod.
 The virus replicates in the cytoplasm and matures by budding into vesicles in the
Golgi region and then released by exocytosis at the cell surface.
 These viruses are labile outside the host.
Classification
The term arbovirus (arthropod-borne virus) is often used to refer to any virus of
vertebrates transmitted by an arthropod. It thus includes in addition to the viruses in
Bunyaviridae, viruses in the families Arenaviridae, Togaviridae, Flaviviridae, Reoviridae
and Rhabdoviridae. The name "arbovirus" is therefore not considered a legitimate
taxonomic term.
There are presently five genera assigned to the Bunyaviridae and these genera
contain serogroups. The genera are:
Bunyavirus - This genus consists of a large number of serologically grouped and
ungrouped viruses. Most are mosquito-borne; some are tick-borne and some show
transovarial transmission. Included of veterinary and human significance are:
Akabane, Peaton and Aino viruses (members of the Simbu serogroup), which cause
disease in sheep and cattle (described below under Akabane Disease).
Cache Valley virus (see below).
California encephalitis virus: Consists of more than a dozen serologically related
mosquito-borne viruses that can occasionally cause encephalitis in humans in the
USA.
La Crosse virus is a strain of California encephalitis virus that was isolated from a fatal
case of human meningoencephalitis in Wisconsin.
Hantavirus
At least 20 serologically related viruses that cause natural infection in small rodents
(including mice). They are transmitted to humans by inhalation often causing fatal
hemorrhagic fever.
Nairovirus
Nairobi sheep disease virus.
Phlebovirus
Rift Valley fever virus.

Bunyavirus
Akabane Disease
(Congenital arthrogryposis-hydraencephaly)
Cause
Akabane virus and probably the Peaton and Aino viruses.
Occurrence
Akabane disease occurs in cattle, sheep and goats mainly in Australia, some Asian
countries, Argentina, South Africa, and the Middle East.
Transmission
The virus is transmitted to susceptible animals by biting-insects, particularly mosquitoes
and midges (Culicoides).
Clinical & Pathologic Features
There are no overt clinical signs in adult animals exposed to these viruses other than an
early febrile response. Fetal infections, which result in deformities of the fetus, may occur
if pregnant cows are infected during the first trimester of gestation. There may be
abortions, stillbirths, mummified fetuses, and premature live births.
Arthrogryposis is the most common deformity noted, affecting a single joint in one limb
to multiple joints in all limbs and the vertebral column. Severely affected animals may
suffer difficult deliveries. Hydranencephaly is noted less frequently.
Diagnosis
 Clinical specimens: Placenta and fetus.
 A presumptive diagnosis is based on the clinical history and gross lesions in
affected fetuses.
 The virus can be isolated by the intracerebral inoculation of young mice and in
various cell cultures.
 Diagnosis is supported by detection of specific neutralizing antibodies in neonates
and aborted calves, lambs and goats.
Prevention
 Killed vaccines are available.
 Akabane disease, although unlikely to occur in North America because of the
reservoir and vectors, is reportable.

Cache Valley Virus Infection

Cache Valley virus has been associated with congenital malformations in sheep. These
defects, which are principally arthrogryposis and hydraencephaly, are virtually
indistinguishable from those observed with Akabane disease. Serologic evidence
indicates that the virus is widely distributed in North America and that numerous
mammalian species may be susceptible to infection.

Nairovirus
Nairobi Sheep Disease
Cause
Nairobi sheep disease virus.
Occurrence
Nairobi sheep disease occurs in sheep, goats and occasionally in humans (with fever and
arthralgia) in East Africa.
Transmission
It is tick-borne (Rhipicephalus appendiculatus).
Clinical Features
The disease is characterized clinically by high fever, depression, anorexia, nasal
discharge, and a severe non-contagious hemorrhagic gastroenteritis. Pregnant animals are
likely to abort. The disease is more severe in sheep with the mortality rate ranging from
30 to 90%.
Diagnosis
 Clinical specimens: Whole blood, acute and convalescent sera, spleen, mesenteric
lymph nodes.
 A presumptive diagnosis is made on the basis of clinical signs and history.
 Confirmation requires isolation and identification of the virus or the
demonstration of a significant increase in antibody levels between acute and
convalescent sera.
 The virus is most easily isolated by the intracerebral inoculation of infant mice.
The virus also grows in a variety of cell cultures.
Prevention
 Dipping to control ticks is widely practiced in endemic regions.
 The disease is considered reportable in countries that are free of it.

Phlebovirus
Rift Valley Fever
Cause
Rift Valley fever virus.
Occurrence
Rift Valley fever occurs in Africa and the Middle East in sheep, goats, cattle, camels,
antelopes and humans. Large outbreaks have occurred in Africa, Saudi Arabia and
Yemen and considerable humans have succumbed.
Transmission
By mosquitoes but probably also by contact.
Clinical & Pathologic Features
Infection is most severe in young animals, and is characterized by a high fever, anorexia,
weakness, and rapid death. Some affected animals may have nasal discharge and
hemorrhagic diarrhea. Adult animals are less severely affected, but pregnant animals are
likely to abort. Cattle are less severely affected than sheep.
The mortality rate may exceed 70% in young animals but is considerably less in adults.
Humans may become infected by mosquitoes and through contact with diseased tissues.
Infections are "flu-like", and can infrequently be severe and fatal.
A consistent and characteristic necropsy finding is severe liver necrosis.
Diagnosis
 Clinical specimens: Liver and spleen.
 A presumptive diagnosis is made on the basis of clinical signs and gross and
microscopic lesions observed in the liver.
 Confirmation requires isolation and identification of the virus. The virus
replicates on the chorioallantoic membrane of chicken embryos and in various
cell cultures.
Prevention
 Modified live virus and killed virus vaccines are used in countries where the virus
is endemic. The modified live vaccine should not be used in pregnant animals.
 Mosquito control reduces the chances of infection. In countries where the disease
does not occur, outbreaks are dealt with by strict quarantine and slaughter.

Bornaviridae
This family is named after the town Borna in Germany where the disease in horses was
first observed about 200 years ago. Borna disease virus is the only species in the only
genus, Bornavirus. The virus has recently received considerable attention as a possible
cause of mood disorders in humans. The host range of the virus is much greater than was
earlier noted.
Viral Characteristics
 The virus is spherical, enveloped and about 90 nm in diameter.
 The genome consists of negative-sense, single-stranded RNA.
 Replication is in the cell nucleus with budding at the cell surface.
  The virus is genetically stable with different isolates showing high levels of
sequence conservation.
 It is labile and its survival time outside the host is probably short.
As mentioned Bornaviridae contains only one genus, Bornavirus, and one species the
cause of Borna disease.

Bornavirus
Borna Disease
What follows is mainly a description of the acute disease in the horse. Similar disease
may also be seen in other species.
Cause
Borna disease virus. Although there is considerable genetic homogeneity, genotypes may
adapt to particular hosts.
Occurrence
The disease in horses is endemic in some regions of Germany and Switzerland. In
recent years infections have been reported in cattle, sheep, goats, monkeys, ostriches, cats
and humans. Serological evidence from humans indicates that the virus is probably
widespread in Europe and North America.
Humans are infected by Borna disease virus and show an antibody response but
not clinical disease. Whether or not these infections can be responsible for some
psychiatric disorders is currently under investigation. Some serological evidence suggests
they may be.
Questions that need to be answered are: What is the prevalence rate in humans?
What is the source or sources of human infections? Is there a natural reservoir? Can
humans transmit the disease in the way horses do?
Transmission
Spread is considered to occur by direct and indirect contact. The virus is present in nasal
and oral secretions and urine.
Clinical & Pathologic Features
Following a protracted incubation period (weeks), the virus causes a
meningoencephalomyelitis of horses with clinical signs that are similar to those produced
by eastern, western and Venezuelan encephalomyelitis. They include depression, shaking
and unsteady gait, and running into obstacles. The non-purulent encephalomyelitis
mainly affects the gray matter of the brainstem and the cerebral hemispheres. Affected
horses usually die.
Intranuclear inclusions may be present in neuronal cells of the hippocampus and olfactory
lobes.
Diagnosis
 Clinical specimens: Brain.
 A presumptive diagnosis is often made on the basis of clinical signs and the
finding of typical eosinophilic inclusions in brain tissue on histopathologic
examination.
 Confirmation requires isolation and identification of the virus, which may be
accomplished by the inoculation of embryonated chicken eggs via the
chorioallantoic membrane or by the intracerebral inoculation of rabbits. The virus
also grows in a variety of cell cultures but without observable cytopathic effects.
  Detection of specific antibodies by ELISA or immunofluorescence assay is
supportive.
 A reverse-transcriptase PCR has been used to detect virus.
Prevention
 Affected and seropositive animal should be segregated.

Orthomyxo viridae

Properties
 Pleomorphic spherical or filamentous virion, diameter 80-120 nm
 Envelope containing hemagglutinin (H) and neuraminidase (N) peplomers, a
matrix protein (M1) on the inner surface and a small number of pores composed
of protein M2
 Nucleocapsid of helical symmetry
 Linear minus sense ssRNA genome, 13.6kb, with 8 segments coding for 10
proteins: 5 structural, 3 associated with polymerase and 2 nonstructural.
 Transcription and RNA replication in the nucleus; maturation by budding from
plasma membrane: capped 5' termini of cellular RNAs cannibalized as primers
for mRNA transcription.
Avian Influenza

Synonym: Fowl plague

Avian influenza viruses occur worldwide, and it seems likely that such viruses are
the ultimate source of influenza viruses of mammals, either by direct transfer or via
reassortment. Outbreaks of severe clinical disease, usually caused by subtypes expressing
H5 and H7determinants, occur periodically in chickens and turkeys.

Host
Various species of wild birds, mainly waterfowl, constitute an important reservoir.
Among domestic birds, chickens and turkeys are the species most likely to develop
disease, but pheasants, quail, guinea fowl and partridges also develop clinical
illness.

Properties
14 known hemagglutinin subtypes and all 9 neuraminidase subtypes are present in
the virus.

Pathogenesis
Spread of influenza virus in tissues is dependent on the type of protease present in
a given tissue and the structure of the viral haemagglutinin molecule. The production of
infectious virions requires cleavage of viral haemagglutinin. In the majority of influenza
A virus subtypes, haemagglutinin cleavage takes place in the epithelial cells of the
respiratory and digestive tracts. Because of the amino acid composition at their cleavage
sites, haemagglutinins of virulent subtypes are susceptible to cleavage in many tissues,
facilitating the development of generalized infection.
Clinical features
The incubation period varies from a few hours to a few days, depending on the
virus dose, the virulence of the strain, and the host species. Infection of chickens and
turkeys with highly virulent viruses is characterized by the sudden onset of high mortality
and also by cessation of egg laying, respiratory signs, excessive lacrimation, sinusitis,
edema of the head and face, cyanosis, especially visible on the combs and wattles, and
diarrhoea.
Diagnosis
Suitable specimens for laboratory examination include tracheal and cloacal swabs,
faeces and pooled samples of organs.
Tissue suspensions are inoculated into 9 to 11 day old embryonated eggs.
Allantoic fluid, harvested after incubation for four to seven days is tested for
haemagglutination activity.
Presence of virus can be confirmed by immunodiffusion using a suspension of
chorioallantoic membrane from eggs inoculated with material from an outbreak and
positive serum.
Haemagglutination inhibition or immunodiffusion tests
The intracerebral pathogenicity index and intravenous pathogenicity index tests,
in one day-old chicks and 6-weeks-old chickens, respectively, are commonly used. If
there is mortality in more than 75% of chicks within 8 days (atleast 8 chicks have to be
used) it is defined as highly pathogenic.
Genomic sequencing

Prevention and control

Initial control is aimed at preventing the introduction of avian influenza viruses.
Once diagnosed, quarantine and removal programmes are implemented.
To minimize secondary spread, strict hygienic measures must be introduced
which include cleaning and disinfection, an interval between slaughter and repopulation,
and controlled movement of people and animals. Local quarantine and depopulation of
affected farms remain the only effective way to eradicate virulent influenza viruses from
chicken and turkey farms.
Vaccination with inactivated vaccines preferably polyvalent vaccine.

Swine influenza

This highly contagious disease of pigs occurs worldwide. Swine influenza was
first described in 1918, its occurrence coinciding with a major pandemic of human
influenza. Two cocirculating subtypes, H1N1 and H3N2, are endemic in pig populations.
In Europe during 1979, H1N1 isolates, clearly distinguishable from classical H1N1
subtypes and with haemagglutinins structurally similar to avian haemagglutinins, were
identified. These H1N1 subtypes, which are more virulent than classical H1N1 isolates,
now predominate in Europe. Acute respiratory disease of pigs in Japan and in the United
Kingdom has been attributed to the H1N2 subtype. There is convincing epidemiological
evidence to support the view that transfer of virulent subtypes from pigs to humans is a
major factor in the emergence of pandemics in the human population.
Epidemiology
An outbreak of swine influenza is usually associated with the recent introduction of pigs
into a herd. Virus shed in high concentrations in the nasal secretions of the infected pigs,
spreads rapidly within a herd. The principal route of transmission is by direct contact.
Airborne spread between farms may occue under suitable weather conditions in areas
with high pig densities. Outbreaks of disease usually occurs when environmental
temperatures are low.

Pathogenesis and pathology

Infection is limited to the respiratory tract; the lungs are the major target organs.
Following infection, virus multiplies in nasal, tracheal and bronchial epithelium. Spread
of infection throughout the respiratory tract results in necrosis, extensive pneumonic
change and consolidation. Lesions are often limited to the apical and cardiac lobes. The
acute phase of the disease persists for more than 72 hours after which virus replication
declines.

Clinical signs
Onset of the disease in a herd is often abrupt, many pigs becoming clinically ill
simultaneously. The incubation period is up to three days. The severity of the illness
ranges from subclinical to acute and is strongly influenced by the strain of the infecting
virus. Secondary bacterial infections frequently complicate the course of the disease and
delay recovery. Acute disease is characterized by huddling in groups, paroxysmal
coughing, dyspnoea and fever. Some pigs may have a discharge from the eyes and nose.
Most pigs recover within six days. Mortality is usually low except in very young pigs or
when intercurrent infection is present. The economic impact of the disease is mainly
attributable to loss of weight. In fully susceptible herds, abortion can occur in affected
sows.

Diagnosis
• Samples suitable for virus isolation include nasal mucus and lung tissue from
acute cases early in the disease. As the virus is labile, transport media should be used for
rapid transfer of specimens to the laboratory. Isolation is usually carried out in
embryonated eggs. After incubation for 72 hours, hemagglutinating activity is
demonstrable in the allantoic fluid.
• Demonstration of a rise in antibody levels in paired serum samples using
haemagglutination-inhibition test or ELISA procedures is indicative of infection.
• Viral antigen can be detected using immunofluorescence or ELISA.
• Viral nucleic acid can be detected with PCR.

Control
Good husbandry, including the elimination of stress factors may help to minimize
losses from swine influenza. Inactivated vaccines are available commercially.
Vaccination can be beneficial provided that the subtypes of virus incorporated in vaccines
include those involved in the outbreaks.
 Equine influenza

This economically important, acute respiratory disease of horses occurs

worldwide except in Australia, New Zealand and Iceland. Two immunologically distinct
subtypes of influenza A virus are described in horses. The virus, first isolated from
horses in 1956, was designated A/equine/Prague/l/56 (H7N7) or influenza A/equine 1. In
1963, a second subtype was isolated in the USA and designated A/equine/Miami/2/63
(H3N8) or influenza A/equine 2. Infection or vaccination with one subtype does not
induce protection against infection with the other subtype. Although the last outbreak of
disease attributed to influenza A/equine I occurred in 1979, there is serological evidence
that this subtype continues to circulate in the horse population.
Antigenic drift accounts for several variants of influenza A/equine 2 with two
antigenically and genetically distinct lineages identified in Europe and the Americas. In
contrast, the H3N8 subtype isolated from horses in China was more closely related to
avian strains than to the H3N8 subtype circulating in horses elsewhere.
Epidemiology
Outbreaks are associated with movement and assembly of horses for shows, sales,
racing or training. The initial source of infection is often a partially immune horse
shedding virus without showing clinical signs. Equine influenza is highly contagious and
spreads rapidly among susceptible horses. Large quantities of virus are shed in aerosols
by the frequent coughing of affected animals. Infection can be acquired at distances up to
30 metres. Indirect transmission through contamination of clothing, equipment and
vehicles can also occur.

Pathogenesis
The virus replicates in the epithelium of the respiratory tract resulting in
destruction of ciliated epithelium and hypersecretion from submucosal glands.

Clinical signs
The incubation period is up to two days. Affected animals develop a high
temperature, nasal discharge and dry cough. Anorexia and depression, although
common, can vary in intensity. Ocular discharge, limb oedema and stiffness may also be
present. Age and previous exposure or vaccination status may influence the severity of
the clinical signs and the likelihood of secondary bacterial infection with the development
of respiratory complications. Exercise exacerbates the clinical signs. Animals with mild
infections usually recover within three weeks. In severe cases, several months may be
required for convalescence.
Diagnosis
Although clinical signs may be suggestive of equine influenza, laboratory
confirmation is required.
• Nasopharyngeal swabs collected during the acute phase of the infection are
suitable for isolation of the virus in embryonated eggs or in cell culture. New isolates
should be closely monitored for antigenic drift.
• A commercial diagnostic kit, developed for the detection of the nucleoprotein of
human influenza A virus, can be used for the diagnosis of equine influenza.
• Serological diagnosis of equine influenza is possible. Haemagglutination
inhibition or single radial haemolysis tests on paired serum samples can be used for
diagnosis. Serum used in the HI test must be pretreated in order to remove non-specific
inhibitors.
Treatment and control
Supportive therapy and rest is indicated for affected horses. Several inactivated
vaccines are commercially available. However, immunity is usually short-lived and
booster injections are required in accordance with manufacturer's instructions. The
incorporation of polymer adjuvants or Quil-A based immune-stimulating complexes
(ISCOMs) into vaccine preparations extends the duration of protective levels of
immunity. Vaccinated horses generally exhibit milder clinical signs and shed virus for
shorter periods than unvaccinated animals. Vaccine manufacturers must update vaccinal
strains regularly. Vaccines should include antigenic material representative of the
influenza A virus subtypes prevalent in the horse population. In addition to vaccination,
control of equine influenza requires isolation of affected horses and cleaning, disinfection
and isolation of infected premises. Animal movement should cease until contaminated
premises have been cleaned and disinfected.

Canine Influenza
(Dog flu)
Cause
Influenza A virus, closely related to subtype H3N8 and presumed to have been acquired
from a horse(s). Subtype H3N8 virus is a frequent cause of equine influenza.
Occurrence
Dog flu first occurred in greyhounds in Florida, USA in January 2004. Of the dogs
affected by the outbreak about 30% died. The disease was first seen in race tracks but has
since spread to veterinary clinics, boarding facilities and animal shelters. Infections have
also been confirmed in New York and a number of other states.
Older dogs and puppies may be more severely affected.
Transmission
This is mainly by infectious aerosols resulting from coughing but also indirectly by
fomites.
Clinical Features
It appears that the disease is less severe than earlier thought and about 80% of those
infected come down with a mild disease. The incubation period is 2 - 5 days. Among the
clinical signs are fever, coughing, dyspnea, anorexia, depression and mucoid to purulent
nasal discharge. In mild cases recovery is within 1 - 3 weeks. Pneumonia with secondary
infection is the most common and serious complication. The fatality rate has ranged from
5 - 8% in the early outbreaks among greyhounds but somewhat less in later outbreaks.
Diagnosis
Dog flu can only be distinguished from other respiratory infections, such as kennel
cough, by laboratory means.
 Specimens: Nasopharyngeal swabs taken within 72 hours of the appearance of
signs; paired serum samples with a three-week interval.
 Virus isolation is readily accomplished in chicken embryos.
 Viral antibodies are identified by hemagglutination-inhibition or virus
neutralization.
  There are several rapid commercial tests for antigen used for the diagnosis of
human influenza which may have application for the canine disease. A rapid
human influenza A kit is used for the diagnosis of equine influenza.
Treatment
 The antiviral drugs oseltamvir (Tamiflu), if used within 48 hours of evidence of
infection, is effective in the treatment of human influenza. There is as yet no
information on its use in dog flu.
 Broad spectrum antibiotics are employed in the severe form to treat secondary
bacterial infections.
Prevention
 A vaccine is not yet available.
Public Health Significance
Transmission of dog flu to humans has not been reported.

Feline Influenza
In 2004, a few tigers and leopards in a Thialand zoo fed poultry carcasses infected
with avian influenza (H5N1) developed a severe, fatal pneumonia as a result of H5N1
infection. Furthermore, in 2005 in the same zoo, horizontal transmission of H5N1
influenza between tigers is thought to have occurred.
It has also been reported that domestic cats can be infected experimentally
(horizontally) with avian H5N1 virus. Cats thus infected developed severe diffuse
alveolar damage and transmitted the virus to sentinel cats.
In 2005 an H5N1 infection was confirmed in a domestic cat in Germany; the
disease was not spread to other cats. It was thought that the infection was acquired from a
wild bird.
These reports suggest that the H5N1 virus should be considered a potential threat
to domestic and captive felids.
It has been suggested that cross-species infection, e.g., from bird to cat requires a
large infective dose of virus whereas in epidemic influenza within a species infection
may result from less than ten viruses.

Arenaviridae

Viruses of this family have a nucleocapsid and a single-stranded, circular RNA in two
segments. None of these viruses is pathogenic for domestic animals but some cause
significant human diseases. Their natural hosts are wild rodents.
Viral Characteristics
 These viruses are enveloped and have a helical nucleocapsid (50 - 300 nm in
diameter) and a single stranded circular, negative-polarity RNA in two segments.
A portion of both segments has positive-polarity RNA (although it is not
translated) and thus the term ambisense is applied to this unusual genome.
 Another distinctive feature is the presence of granules (nonfunctional ribosomes)
on the viral surface.
 They replicate in the cytoplasm and mature by budding through the cell
membrane.
Significance
The Arenaviridae consists of two genera, Arenavirus and Deltavirus, which are discussed
below with significant viruses:

Arenavirus
In rodents, most arenaviruses cause subclinical, lifelong persistent infections with
continuous excretion of virus in saliva, urine and feces thus facilitating spread. Human
exposure is usually occupational and more frequently involves farm workers in rural
areas. Human infection may range from inapparent, mild disease to the severe and
frequently lethal hemorrhagic fevers. More than 20 species of arenavirus species have
been identified worldwide. All have rodents as reservoirs and some cause human disease.
The prototype of this family is the lymphocytic choriomeningitis virus (LCMV), a
virus found to infect wild rodents, mice and occasionally humans. Immunological studies
employing LCMV have contributed significantly to our knowledge of tolerance, virus-
induced immunopathology, viral antigen recognition by T lymphocytes, natural killer
activation and function, among others. Lymphocytic choriomeningitis virus is a rare
cause of aseptic meningitis in humans.
Arenaviruses causing human disease must be handled in laboratories under strict
containment conditions (Biosafety level 4) to prevent human exposure.
Lassa fever virus causes a frequently fatal hemorrhagic fever in humans in West Africa.
The natural host is the field rat.

ORDER: NIDOVIRALES

The order nidovirales encompassing two families: the family Corona viridae with
two genera, corona virus and torovirus, and the family Arteri viridae, with one genus,
Arteri virus.
The name nidovirales derived from the Latin nidus, which means nest. Despite
differences in virion structure and genome size corona virus, toro virus and arteri virus
exhibit similarities in genome organisation and replication strategy. In replication during
transcription, from negative sense complementary RNA, instead of full length of genomic
RNA, nested sets (5-7) of subgenomic RNA’s will produce.

Properties
SS RNA, positive sense, enveloped virus, replicate in cytoplasm, helical
symmetry. They are roughly spherical in shape, 80-120 nm in size (corona viruses) or
120-140 nm in sizeand disc, kidney or rod shaped (toro viruses). The genome is being the
largest RNA virus genomes known. The corona virus has 20nm long large club shaped or
petal shaped peplomers. These projections are spaced widely apart and dispersed evenly
all over the surface providing a crown like appearance and hence these viruses are known
as corona (means crown) viruses. The 5’ end of the genome is capped and 3’end is poly
adenylated.

The virus has three or four structural proteins:

Peplomer/ surface/ spike glycoprotein S –is responsible for attachment to cells,
and membrane fusion.
 Transmembrane glycoprotein M - responsible for virus envelope
Nucleocapsid phosphoprotein N
Hemagglutinin plus esterase protein, HE - responsible for receptor binding,
haemagglutination and destroying activities.

Replicates in cytoplasm; full-length minus sense RNA strand is transcribed from

the virion RNA, from which a nested set of mRNAs is produced with unique sequences at
their 5' ends, which are translated; maturation is by budding into endoplasmic reticulum
and Golgi cisternae, with virions released by exocytosis.

Family: CORONAVIRIDAE
(Corona = crown)
Genus: Corona virus
Properties
 Pleomorphic spherical virion, 75-160 nm (average 100 nm) in diameter
 Envelope with large, widely spaced, club-shaped peplomers
 Tubular nucleocapsid with helical symmetry, 10-20 nm in diameter
 Linear plus sense ssRNA genome, 27-33 kb, capped and polyadenylated,
infectious
 Three or four structural proteins : Peplomer glycoprotein S (E2, 180K-20K),
Transmembrane glycoprotein M (EI-23K-29K), nucleocapsid phosphoprotein N
(50K-60K); some viruses have peplomers with hemagglutinin plus acetylesterase
activity, HE (E3, 62K-65K).
 Replicates in cytoplasm; full-length minus sense RNA strand is transcribed from
the virion RNA, from which a nested set of mRNAs is produced with unique
sequences at their 5' ends, which are translated; maturation is by budding into
endoplasmic reticulum and Golgi cisternae, with virions released by exocytosis.

Basis of genetic and serologic properties, the genus corona virus can be sub
divided into three groups.

Antigenic groups and Diseases caused by corona viruses

Antigenic group Virus Disease
Group –1 Transmissible gastro enteritis virus Gastro enterits
(Mammalian of swine
viruses) Feline infectious peritonitis virus Peritonitis, pneumonia,
menigioencephalitis,
wasting syndrome
Canine corona virus Enteritis
Group –2 Porcine haemagglutinating Vomiting, wasting and
(Mammalian and encephalomyelitis virus encephalomyelitis
avian)
Blue comb virus of turkey Enteritis

Bovine corona virus Gastro enteritis

(Winter dysentery)
Group –3 Infectious bronchitis virus Tracheobronchitis,
( avian) ( atleast eight serotypes) nephritis

AVIAN INFECTIOUS BRONCHITIS

Synonym: Gasping disease

An acute, highly contagious respiratory disease of chickens characterized by

respiratory signs such as coughing, sneezing, tracheal rales with accumulation of mucus
in the bronchi and depression, heavy mortality in young chickens, stunting growth,
nephritis, increased feed convertion ratio, long –term reproductive problem, decreased
egg prodution and affect both internal and external quality of egg.

Properties
Virion properties are similar to family. The IBV is sensitive to ether treatment,
complete inactivation not achieved. Susceptible to temperature, pH and chemicals varies
with different strains. Readily inactivated at temperature of 56 0C for 15 mts. Vrus in
allantoic fluid can be kept at –200C for few months. 10% glucose/ 50% glycerine are used
as stabilizer.

IBV will HA activity after the virus treated with any one of the enzymes such as
phospholipase-C, Neurminidase and trypsin

Antigenic property
The virus elicits distinct antigen determinants on envelope and spikes. Variations
within the virus are due to differences in the S and HE proteins. As a result of this there
are many serotypes and within each serotype there are many strains. These serotypes are
immunologically distinct and antibodies against one serotype may not protect the birds
against the infection caused by other serotypes. Some of the common serotypes are
Massachusetts, Connecticut, and Arkansas etc. some of the strains are MA5, M48, M41,
H120, H151, Holte, Gray, Florida etc.

Cultivation
IBV is cultivated in embryonated eggs and tracheal organ culture system. This
virus grows very well in allantoic route of 9-11 day old chicken embryos. After 3-4
passages, the virus will produce characteristic curling and dwarfing of embryo. Urate
deposits in embryonic mesonephros and allantoic fluid also seen. The allantoic fluid
should not agglutinate red blood cells until treated with enzyme.

Tracheal oragn culture systems prepared from 20-day old embryos can be used to
isolate IBV directly from field material. Tracheal rings of 0.5-1.0 mm thick are prepared
using automatic tissue chopper. Infection of tracheal organ cultures produce ciliostasis
within 24-48 hrs.

In chicken embryofibroblast, chicken embryo kidney and chicken embryo lung

cell cultures the IBV produce syncyita and plagues with intra cytoplasmic inclusions.
Epidemiology
IB is mainly a disease of chickens. Pheasants, pigeons and guinea fowl are also
affected. All ages, both broilers and layers are susceptible. IBV is the most rapidly
spreading virus known in birds. Virus was isolates from respiratory tissue, oviduct and
eggs after 50 days of post infection and 20weeks post infection in faeces. Site of virus
persistence is kidney. Affected chicks expelled large quantities and high titre of virus in
respiratory secretions. In the environment the virus can survive on fomites for several
days and possibly for weeks, especially at low environmental temperatures. Virus can
travel several miles through air. The most important route of transmission is by aerosols
and spread of infection also occurs by ingestion of food contaminetd with faeces.

Pathogenesis
The virus replicates to high titre first in the respiratory tract (ciliated epithelial
cells); this is followed by viremia (within 1 to 2 days of infection), which distributes the
virus to many organs. Eventually the virus causes major damage to the reproductive
system and the kidneys. The intestinal tract is another site of primary infection. The virus
becomes widely distributed throughout the body, particularly in the oviducts, kidneys and
bursa of Fabricus.

IBV has unique ability among avian virus of producing permanent anatomical
change to the immature oviduct.

Carrier status may also be seen in some birds and these birds remain persistently
infected and shed the virus in feces for many months.

Clinical Features
Incubation period is 18 to 36 hours. Age, immune status, time and route of
infection, and strain of the virus strongly influence the nature and severity of disease
observed in a flock. Chicks between 1 and 4 weeks of age show the most severe disease,
which is characterized by gasping, coughing, rales, nasal exudates, and respiratory
distress. Virus spreads very quickly and the entire flock gets affected within a short span
of time. Chicks demonstrate high-pitched cheeps, become lethargic, loss appetite, and
may die suddenly. The course of the disease is 7-21 days. Mortality usually 25-30 % but
sometimes reaches 70-80%.

In adult birds the disease may go unnoticed. But the layers show respiratory rales
and coughing with prominent involvement of the reproductive tract and a decline or
cessation of egg prouction. When laying is resumed, many eggs show watery albumen,
no shells, thin shells, shells with deposits, distortions, depression or ridging. There is loss
of albumen quality, sometimes the thick white disappearing almost completely. Strains of
low virulence cause pasting i.e. the accumulation of white, sticky exudates around the
vent that can plug the cloaca.

Infection with nephrotropic strains (e.g. Holte and gray strains) of IBV is
associated with interstitial nephritis and mild respiratory signs with moderate to high
levels of mortality. Mortality in renal form may goes upto 60%. Nephritis is an immune
complex disease. It is occur mainly due to the long time persistence of virus leads to the
production of high level of antibody.

Lesions
Loss of ciliary epithelium, thickening of mucus, swollen sinuses, tracheitis,
bronchitis, air sacculitis and conjunctivitis are seen. Yellowish caseous dry exudates
block the main bronchi or trachea may be the main cause of death.

In layers, the ova can be congested and sometimes ruptured, with free yolk in the
abdominal cavity. Kidney gets enlarged to 4 or 5 times more than normal size and urate
deposits are seen.

Diagnosis
Samples to be collected
Live birds: Scoop the tracheal mucus by sterile scalpel and faeces.
Dead birds: Tissue samples from kidney, oviduct, spleen and bursa

Isolation and identification of virus

The samples are inoculated into allantoic route in 9 to 11 day old embryos
obtained from seronegative hen. By means of blind passage- mortality rate increases as
the passage level increases which kills embryo within 48 hrs, "Dwarfing and curling of
embryo" is very characteristic. HA test and ciliastais in tracheal organ culture system are
also helpful

By serology
Serotype indetification by HI and VNT are highly essential.
Demonstration of viral antigen in tracheal tissue smear by IFT is highly valuable.
AGPT, CIE, SNT, ELISA and RT-PCR are highly valuable in detection of antigen.

Differential diagnosis
IB should be differentiated from ND, ILT and Infectious Coryza.

Prevention and control

IB is difficult control because of persistent infection. Attenuated virus vaccines
are widely used. These vaccines are derived either from avirulent field isolates or by
passage in embryonated eggs. They are administered in drinking water, by coarse spray,
or by parenteral route. Vaccine is usually given between 7 and 10 days of age, again at 4 th
week and 12th week by using M41/ M48/ MA5/ H120. Day old vaccination or at 7 th day of
age is followed in broilers.

Note: Vaccination breaks are common because of the variable presence of new antigenic
variants. Such variants will continue to emerge and spread, posing continuing problems
for poultry producers.

BLUE COMB DISEASE

 The disease affects turkeys of all ages but is most severe in I to 6 week old poults.
Only one serotype of blue comb virus is recognized. The disease is characterized by loss
of appetite, constant chirping, diarrhea, weight loss and depression. The skin of head and
neck may become cyanosed. An inactivated vaccine is available, but it is generally
considered to be ineffective. Young poults usually die.

TRANSMISSIBLE GASTROENTERITIS

Transmissible gastroenteritis (TGE) is a highly contagious, coronaviral disease of

young pigs, which occurs worldwide. There is one serotype of transmissible
gastroenteritis virus (TGEV), which is closely related antigenically to feline coronavirus
and canine coronavirus.

Epidemiology
Transmission of TGEV is usually by the faecal-oral route. The virus is moderately
stable in the presence of proteolytic enzymes and at pH 3.0, ensuring survival in the
stomach and small intestine. Viral shedding in faeces can persist for up to two weeks.
Outbreaks of TGE tend to occur in winter. In fully susceptible herds, the virus spreads
rapidly infecting animals of all ages. The disease, however, is most severe in newborn
piglets. Outbreaks usually terminate in a few weeks if no new susceptible animals are
introduced into the herd.

Pathogenesis
Following ingestion, the virus replicates mainly in mature enterocvtes at the tips
of the villi on small intestine. Viral replication results in villus atrophy throughout the
length of the small intestine. Digestion and cellular transport of nutrients and electrolytes
are severely disrupted resulting in the accumulation of fluid in the intestinal lumen and
diarrhoea. Young piglets are particularly susceptible to the ensuing dehydration and
metabolic acidosis.

Clinical signs
The incubation period is up three days. Vomiting and watery diarrhoea may be
evident in affected piglets less than seven days old. Rapid dehydration and weight loss
follow. Mortality may approach 100% in newborn piglets and is usually confined to
animals under three weeks of age. Inappetence and transient diarrhoea may be observed
in older pigs. Subclinical infections also occur. Sows quickly become immune and
maternally derived immunity reduces the severity of clinical signs in piglets. Outbreaks
usually last a few weeks. However, TGEV infection may become endemic in a herd if
consecutive litters become infected as maternally derived immunity wanes. Clinically,
such infections are usually mild.

Diagnosis
The sudden onset and rapid spread of diarrhoea among newborn pigs along with
almost 100% mortality is highly suggestive of TGE. Postmortem examination of washed
small intestine discloses paper-thin walls, due to villous atrophy. The walls of the
jejunum and ileum are affected while those of the duodenum are usually normal.
• Viral antigen can be detected in mucosal smears or cryostat sections of the small
intestine by immunofluorescence. Viral antigens can be demonstrated in faeces by
ELISA.
• Virus can be isolated from faeces in a swine testis cell line.
• Serological testing for antibodies can be carried out using virus neutralization.
TREATMENT AND CONTROL
• There is no specific treatment but fluid replacement therapy may be beneficial.
Maintaining the farrowing house at an optimal temperature may enhance survival.
• In acute outbreaks of TGE, deliberate exposure of pregnant sows to the virus
may reduce neonatal mortality. After exposure, sows due to farrow should be moved to
clean premises. Newborn piglets born to exposed sows will usually receive passive
antibody protection following suckling.
• Modified live and inactivated vaccines are available. Modified live vaccines are
administered orally to sows five to seven weeks before farrowing and a booster
inoculation is administered parenterally one week before parturition. Vaccination reduces
mortality but does not eliminate infection.

Porcine Epidemic Diarrhea

Cause
Porcine epidemic diarrhea virus.
Occurrence
Porcine epidemic diarrhea virus affects swine in Asia and Europe, but not in the
Americas.
Transmission
Infection is by the oral or nasal route. Spread is by direct or indirect contact.
Clinical & Pathologic Features
The virus infects the epithelial cells of the villi of the small intestine, causing
histopathologic lesions similar to but milder than those of TGE.
The incubation period is about 1 - 4 days. The principal clinical sign is watery diarrhea
and all ages of pigs may be affected. The clinical disease is somewhat similar to TGE but
differs in that spread of the virus throughout the herd is much slower, vomiting is not a
predominant clinical sign, and mortality is lower. In addition, older pigs are occasionally
more severely affected than younger ones.
Diagnosis
 Clinical specimens: Portions of small intestine with contents.
 The virus can be propagated in cell cultures if trypsin treatment (to cleave the
fusion protein enabling the virus to infect the next cell) of inoculum is utilized
between transfers.
 Diagnosis is usually accomplished by fluorescent antibody examination of
cryostat sections of affected intestine or by immune electron microscopic
examination of feces or intestinal contents.
 Paired serum samples are tested for antibodies with an ELISA or an indirect
immunofluorescence assay.
Prevention
 Vaccines are not available.
 Strict sanitary measures to prevent spread, along with deliberate infection of
pregnant sows have reduced losses.

Feline Infectious Peritonitis

Cause
Feline coronavirus.
Occurrence
This frequently occurring, widespread disease affects domestic and wild felidae. The
morbidity rate is low, and although a number of cats in a household or colony may be
affected, most cases are sporadic.
Transmission
The virus is present in blood and exudates of infected animals. Transmission requires
close contact with carrier or clinically infected cats, and is thought to occur by the oral
route and through inhalation of aerosol droplets.
Pathogenesis
Infection is thought to begin in the intestinal epithelium and regional lymph nodes with
spread to target organs via infected macrophages. Development of disease depends on the
degree and nature of pre-existing immune responses. Two forms of the disease, "wet" and
"dry", are described but a definite distinction between them cannot always be made. It is
suggested that the cell-mediated response accounts for the dry or non-effusive form,
while the wet or effusive form develops in the absence of a sufficient cell-mediated
response. Antigen-antibody complexes are considered responsible for the lesions in the
wet form.
Clinical & Pathologic Features
Feline infectious peritonitis (FIP) is a progressive debilitating febrile disease affecting
cats of all ages, although most cases occur in cats six months to two years of age.
The incubation period may be as short as two weeks or as long as several months.
In the wet or effusive form, the initial clinical signs consist of anorexia, high temperature,
and depression followed by progressive emaciation. The abdomen is often enlarged as a
result of the accumulation of fibrinous fluid. There is a pyogranulomatous vasculitis due
to precipitation of immune complexes associated with serous membranes. Dyspnea is a
common sign when fluid accumulates in the thoracic cavity.
In the dry form, there is little or no fluid buildup and a febrile response may be the only
initial overt sign of infection. Ocular involvement (anterior uveitis) and CNS dysfunction
are more commonly seen with the dry form. Both forms are generally fatal, but the wet
form proceeds much more rapidly. Experimentally, a more rapid and severe development
of clinical disease (accelerated FIP) occurs in cats with preexisting FIP antibody. This
form has been attributed to the enhancement of viral infection mediated by Fc receptors
present in macrophages ("antibody-dependent enhancement", ADE).
There is considerable variation in the virulence of viral isolates, and most cats with
serologic evidence of FIP never develop the disease. However, a confirmed diagnosis is,
with rare exception, eventually fatal. Most cats with clinical FIP die several weeks to
months after diagnosis.
 The very closely related feline enteric coronavirus (FeCV) is endemic in cats in
which it may cause a mild enteric infection. Evidence suggest that virulent FIP viruses
arise as mutants of FeCV. The FIP virus is also closely related to transmissible
gastroenteritis virus of swine and canine coronavirus.
Diagnosis
 Clinical specimens: Abdominal and thoracic fluids, lung, kidney, liver, spleen and
brain.
 The disease is often diagnosed by gross and histologic examination. In the wet
form, there are varying quantities of a characteristic straw-red fluid with fibrin in
the abdominal and/or thoracic cavities and fibrinous adhesions are present on
visceral organs, particularly the spleen and liver. Multifocal grayish-white
granulomatous-like lesions are usually noted on the surfaces of various organs.
These granulomatous-like lesions are usually larger and penetrate the tissue more
deeply in cats with the dry form of FIP. Fibrinonecrosis and pyogranulomas are
observed upon histological examination. The dry form is difficult to diagnose
clinically. Lesions most commonly affect the eyes, brain, liver and kidneys. X-ray
or ultra sound can be used to detect small amounts of fluid in the chest or
abdominal cavities.
 Virus infected cells can be demonstrated in cryostat sections of affected tissues by
immunofluorescence. Infected cells are usually restricted to or near the surface of
affected tissues, and in and around the granulomatous lesions. Failure to sample
appropriate areas may result in false negative results.
 Ante-mortem diagnosis is difficult and particularly so in the dry form.
Hyperproteinemia is suggestive. Positive immunofluoresence results on cells
collected from effusions are definitive.
 Serologic test (ELISA, indirect immunofluorescence assay) results are considered
of little value in diagnosis, although it is claimed by some that high titers are
suggestive of FIP.
Prevention
 Vaccines are available but their value according to many practitioners is
questionable. They are probably of no value in cats already exposed.
 Disinfection of premises and isolation of seropositive cats. Only admit
seronegative cats.

Canine Coronavirus Infection

Cause
Canine coronavirus. Because the virus can be recovered from many normal dogs, some
investigators have questioned its pathogenicity. Most dogs from shows and in kennels
have antibodies.
Occurrence
Canine coronavirus is worldwide in distribution. The virus is highly contagious, affecting
dogs of all ages, but the disease in puppies is more severe.
Transmission
Virus is shed in the feces and infection occurs mainly by ingestion. Transmission is by
direct and indirect contact.
Clinical & Pathologic Features
Initially, clinical signs are anorexia, depression, and loose stools, followed by vomiting
and diarrhea. Feces often contain mucus (seldom blood) and have a fetid odor. Affected
puppies may become dehydrated.
Asymptomatic infections are common, especially in older dogs. Recovery usually
occurs in 1 - 2 weeks and fatal infections in uncomplicated cases are rare.
Gross necropsy lesions are those of a nonspecific enteritis; microscopic lesions consist of
atrophy and fusion of intestinal villi.
Diagnosis
 Clinical specimens: Fresh feces and intestine.
 Diagnosis is best accomplished by the electron microscopic demonstration of
coronavirus in feces or by the fluorescent antibody staining of cryostat sections of
intestine.
 The virus, which is antigenically related to transmissible gastroenteritis of pigs
and feline infectious peritonitis, can be isolated in cell cultures of canine origin.
 Histopathologic lesions of atrophy and fusion of intestinal villi are suggestive.
Prevention
 Inactivated vaccines are available, usually in combination with other viral and
bacterial agents. Puppies are generally vaccinated twice during the first 1 - 2
months of life and thereafter on a yearly basis.
 Sound sanitation and the use of effective disinfectants (sodium hypoclorite) help
control the disease in kennels.

Feline Coronavirus Infection
Feline coronavirus causes subclinical infections in cats and mild gastroenteritis in kittens.
Feline infectious peritonitis virus is considered to have derived from strains of feline
coronavirus.

Bovine Coronavirus Infection

Cause
Bovine coronavirus.
Occurrence
Bovine coronavirus infection occurs in cattle throughout the world.
Transmission
Virus is shed in the feces. Spread is by direct and indirect contact. The mode of infection
is ingestion. Dams may carry the viruses and infect neonatal calves.
Clinical & Pathologic Features
Coronavirus affects calves in the first three weeks of life. Another virus of bovine
neonates, rotavirus, is seldom found in calves over 7 days of age. Both viruses produce
disease characterized clinically by profuse, watery diarrhea of sudden onset leading to
extreme dehydration. Morbidity is often near 100% and mortality may range from none
to >50%.
Damage to the intestinal mucosa including loss of villous absorptive cells results
in malabsorption. Bovine coronavirus has also been incriminated as a cause of "winter
dysentery" in adult cattle.
Diagnosis
 Clinical specimens: Fresh feces and segments of intestine including spiral colon,
ileum, and jejunum.
 There are several convenient and rapid methods used to diagnose rotavirus and
coronavirus infections, including the electron microscopic examination of
distilled water lysates of feces and fluorescent antibody staining of fecal smears
and frozen sections of intestine.
 Commercial kits (ELISA and Latex agglutination) are available commercially for
detection of rotavirus.
 Both rotaviruses and coronaviruses can be cultivated in cell cultures of bovine
origin but often with some difficulty.
Prevention
 Commercial vaccines are available for bovine rotavirus and coronavirus but their
efficacy is questionable. These may be administered to heifers or cows before
mating.
 Sound management practices including cleaning, disinfection and the use of
footbaths, and provision of colostrum.

Porcine Hemagglutinating Encephalomyelitis Virus Infection
(Vomiting and Wasting Disease)
Cause
Porcine hemagglutinating encephalomyelitis virus. Only one serotype has been identified.
Occurrence
Porcine hemagglutinating encephalomyelitis virus (PHEV) infection is widespread in
North America and Europe.
Transmission
Spread is by contact and aerosol droplets.
Pathogenesis
The virus first replicates in the tonsils and epithelial cells of the upper respiratory tract,
followed by spread via peripheral nerves to the central nervous system. Damage to the
vagal ganglion results in gastric irregularities including vomitting.
Clinical & Pathologic Features
Swine less than two weeks old are more susceptible, but most infections are subclinical.
Clinical disease that occurs in some pigs consists of anorexia, rapid loss of weight, and
depression. Vomiting and constipation may follow. Neurologic signs, such as muscle
tremors, incoordination, and paddling movements, may develop later in the course of the
disease.
Mortality rate may reach 100% in baby pigs.
Diagnosis
 Clinical specimens: Tonsil, lungs, stomach, small intestine, brain, spinal cord, and
acute and convalescent sera.
 Clinically PHEVI may resemble some of the neonatal diseases of pigs, such as
TGE, colibacillosis, and clostridial enterotoxemia; the absence of diarrhea and the
irregularity of vomiting may help in differentiation. The neurologic signs may be
confused with pseudorabies, hog cholera,erysipelas and salt poisoning.
  Laboratory diagnosis is required to confirm PHEV infection. This is most
conveniently accomplished by fluorescent antibody staining of frozen sections of
tissue (brainstem) and virus isolation.
 The virus can be isolated in early passage of porcine cell cultures in which it
produces syncytia.
 Hemadsorption can be demonstrated on infected cells. The virus agglutinates
chicken, sheep, rabbit, rat, mouse, and hamster erythrocytes.
 Histopathologic examination of brainstem may reveal non-suppurative
encephalitis.
Prevention
 The virus is widespread and most infections are subclinical.
 There is no vaccine available and no control measures are practiced.

Arteriviridae
This family of enveloped, positive-sense, single-stranded RNA viruses, established in
1996, was classified formerly in the Togaviridae family. It has only one genus,
Arterivirus, whose virus species are antigenically distinct from each other.
Viral Characteristics
 These positive sense, single-stranded RNA viruses are medium in size (50 - 70
nm) have a spherical appearance due to the envelope, but the nucleocapsid is
icosahedral in shape.
 They possess a lipoprotein envelope with ring-like structures on the surface, but
no gross surface spikes.
 They replicate in the cytoplasm of macrophages and endothelial cells.
 The genome is 13K nucleotides in length, has a 5'-methylguanosine cap and a 3'
poly A tail of approximately 50 nucleotides. The genome alone is infectious.
 Individual viruses are antigenically distinct and host specific; they establish
persistent infections.

Classification
There is only one genus:
 Arterivirus
o Equine viral arteritis virus
o Porcine respiratory and reproduction syndrome virus
o Lactate dehydrogenase-elevating virus of mice
o Simian hemorrhagic fever virus: Causes hemorrhagic fever in several
species of African monkeys.

Arterivirus
Equine Viral Arteritis
Cause
Equine arteritis virus.
Occurrence
Hosts are horses, donkeys and mules. Equine viral arteritis (EVA) is an important
contagious disease occurring worldwide. Outbreaks have frequently been associated with
racetracks. The virus is said to be endemic in standard-breds.
Transmission
The virus is shed in excretions/secretions and transmission occurs by direct and indirect
contact, including by breeding. The most common route of infection in young animals is
thought to be respiratory. Aerosol spread and transmission occurs mainly when horses are
congregated at sales, racetracks and shows.
Pathogenesis
Following infection by the respiratory route the virus replicates in pulmonary
macrophages and then in bronchial lymph nodes. There is a viremia with infection of
endothelial cells leading to a general necrotic arteritis.
Clinical & Pathologic Features
Horses infected with EVA virus may have no overt signs of disease or may display a
variety of clinical signs, including fever, depression, anorexia, and nasal and ocular
discharge. Affected horses often have hind limb and scrotal edema and some develop a
skin rash over various parts of the body. Fatal cases with pulmonary edema and
interstitial pneumonia have been described in foals. Mares are likely to abort. Aborted
fetuses may only show autolysis. Infected stallions recover clinically, but often remain
persistently infected and spread the virus during natural breeding or artificial
insemination.
Gross lesions include edema, congestion and hemorrhages particularly in the
subcutaneous tissues of the abdomen and limbs and widespread necrotic arteritis.
Diagnosis
 Clinical specimens: Nasal swabs, ocular swabs, whole blood, serum, fetal tissues,
and semen from suspect stallions.
 Diagnosis is usually made on the basis of virus isolation or by the demonstration
of a significant increase in specific antibody between acute and convalescent sera.
 The virus can be propagated in a variety of cell cultures, including those derived
from horses and rabbits.
 The serologic test most often used to measure antibody response is virus
neutralization, performed in the presence of 10% guinea pig complement. Other
serological procedures are used but they are thought to be less reliable.
Prevention
 A modified live vaccine given at 6 - 12 months is effective but it is not used in
pregnant mares.
 Prevention is best accomplished by isolation and quarantine of new additions and
horses returning from race tracks and shows.
 Serologically positive stallions should be evaluated as to carrier status through
virus isolation attempts on the sperm-rich fraction of semen or by a program of
test breeding of seronegative mares.
 Pregnant mares should be isolated from other horses.

Porcine Reproductive and Respiratory Syndrome

Cause
Porcine reproductive and respiratory syndrome virus. Antigenic differences have been
noted among different isolates. Three different genotypes have been identified recently.
Occurrence
The virus of porcine reproductive and respiratory syndrome (PRRS) has been reported
from many countries but not all. It has not been reported from Brazil and Argentina.
Transmission
By aerosol, contact, and fomites. Infected boars shed virus in their semen.
Clinical & Pathologic Features
Most infections occur by the respiratory route with initial viral replication in alveolar
macrophages.
The incubation period is typically 2 - 7 days. Clinical signs in adult swine are usually
nonexistent to mild, consisting of a slight febrile response and anorexia of short duration.
A bluish discoloration of the ears due to erythematous plaques has been associated with
PRRS. Pregnant sows may abort late in gestation or deliver prematurely. Abortions
and /or stillbirths may reach epidemic proportions in fully susceptible herds. Young pigs
infected with PRRS virus frequently exhibit depression, anorexia, and rapid respiration
with some coughing and sneezing. Mortality may be as high as 50% in nursing pigs, but
is generally low in older pigs. Secondary bacterial infections may result in poor growth
and performance.
Gross necropsy lesions are minimal in the uncomplicated respiratory form of PRRS, but
interstitial pneumonitis is a consistent histopathologic finding. There are no gross or
histopathologjc lesions noted in aborted or stillborn fetuses.
Diagnosis
 Clinical specimens: Nasal swabs, blood, serum, lung tissue, and aborted fetuses.
 The PRRS virus can be cultivated in porcine alveolar macrophages and in a
continuous cell line derived from African green monkey kidney cells.
Identification is most easily accomplished by indirect fluorescent antibody (IFA)
examination of infected cultures. IFA is also used to detect specific anti-PRRSV
antibody.
 Infection can also be diagnosed by using the IFA test on frozen sections of lung
and fetal tissues.
 Demonstration of seroconversion in affected pigs or in sows that have aborted is
also employed.
Treatment
Appropriate antimicrobial therapy may be indicated to cope with secondary bacteria.
Prevention
 This is best accomplished by good management practices, including the
quarantine and serologic testing of replacement stock and show animals.
 The possibility that previously infected boars might shed virus in their semen for
extended periods of time should be considered.
 A modified live vaccine is given to sows prior to breeding and to pigs 3 - 20
weeks of age. More effective and safer vaccines are currently under development
and may be available in the near future.

Lactate Dehydrogenase-Elevating Virus of Mice

Although of little veterinary significance, this agent illustrates the remarkable capacity
some viruses have for adaptation and survival. Wild and laboratory mice infected with
this virus remain persistently infected for life. The virus infects macrophages and there is
a life-long viremia. They have no overt signs of illness but plasma lactic dehydrogenase
levels are elevated, not as a result of increased production but as a consequence of
impaired clearance. Mice are most often infected during fighting and experimental
procedures, such as the use of common needles, transplantation studies, etc. Natural
transmission appears to require close contact with contaminated blood or tissue, and is
more common in males that fight.

PICORNAVIRIDAE

Picornaviruses have played an important role in the history of virology and in the
history of veterinary medicine. 1n 1897, Loeffler and Frosch first demonstrated that a
disease – FMD - of animals was caused by a filterable virus. A Century later the same
virus was among the first animal viruses to have its structure resolved at the atomic level
by x-ray crystallography.

Properties
The family Picornaviridae is divided into six genera; Aphthovirus, Enterovirus,
Cardiovirus, Rhinoviurs, Hepatovirus, and Parcheovirus. An important difference
between viruses of the six genera is their stabiliy at low pH. The aphthoviruses are
unstable below pH 7, the rhinoviruses lose infectivity below pH5, and the enteroviruses,
hepatoviruses, cardioviruses, and parcheoviruses are stable at pH3.

Picornaviruses of animals

Genus Virus Principal Species Disease

affected
Foot-and-mouth disease Cattle, Sheep, FMD in ruminants and
viruses A, O, C SAT1, Goats, Swine, swine
Aphthovirus SAT2, SAT3, Asia ruminant wild life
species
Equine rhinovirus 1 Horses Resp. signs
Swine vesicular disease Swine Swine vesicular disease
vrius
Entero virus Porcine enterovirus-1 Swine Polioencephalomyelitis
Avian entero viruses Chickens Avian
encephalomyelitis
Ducks Duck viral hepatitis
Cardio virus Encephalomyocarditis Swine, elephants, Rarely,
virus other mammals encephalomyocarditis in
swine and elephants
Rhino virus Bovine rhino viruses Cattle Mild rhinits
1-3

Virion Properties
Virions are non enveloped, 27nm in d.m., and have icosahedral symmetry. The
genome consists of a single molecule of postive sense, ssRNA. The genomic RNA is
polyadnylated at its 3’ end and has a protein, VPg, linked covalently to its 5’ end.
Genomic RNA is infectious. Sixty capsomers are constructed from 60 copies of each of 4
capsid proteins, VP1, VP2, VP3 (M.W. 30,000 each) and VP4 (M.W. 7000 – 8000). VP1,
VP2, VP3 are structurally rather similar to one another differing primarily in the size and
conformation of the loops. Aminoacid substitution correlating with antigenic variation
ocuur in the surface oriented loop regions. The VP1 proteins are located around the five
fold axes of icosohedral symmetry, and VP2 and VP3 alternate around the two-and
threefold axes. VP4 is located entirely at the inner surface of the capsid, probably in
contact with the RNA.

In poliovirus and rhinovirus virions the packing together of VP1, VP2 and VP3
results in the formation of a canyon. The conserved aminoacids on the floor of the canyon
are believe to form the points of attachment of the viruses to cell surface receptors and it
was proposed that this location might shield attachment sites from immune surveillance,
because antibody molecules can not fit into the canyon.
Rhinoviruses and FMDV have a comparatively smooth surface with no canyon
structure; attachment site for host cell receptors are located at the tips of protrubences on
the virion surface. These sites are strongly antigenic and they also have serotype and
subtype antigenic specificities that differ among various FMD viruses.

Virions replicate in the cytoplasm. Virion RNA acts as mRNA and is translated
into a polyprotein which is then cleaved to yield 11 individual proteins.

FOOT AND MOUTH DISEASE

Synonym : Aphthous fever, Epizootic aphthea, Aftosa, Fast Moving Disease

FMD is an acute, febrile, highly contagious disease of almost all cloven hoofed
animals characterized by the formation of vesicles (fluid-filled blisters) and erosions in
the mouth, nose, teats and feet with high morbidity and low mortality.

Importance of FMD
 Very short incubation period
 Rapid replication cycle 3 to 10hrs. (Eclipse period 3hrs)
 The virus shed in all excretions even 24 hrs before onset of symptoms
 Virus spread very fast by windborne
 High morbidity (100%) and Low moratlity (2% in adult & 23 % in young)
 Recovered cattle act as permanent carrier
 Drop in milk production, droughting power, growth of beef cattle, semen
quality never return to normal
 Approximately 5000 outbreaks of FMD occur every year in INDIA results
in loss of Rs. 500 crores per annum (NDDB, 1976).
History
Worldwide the disease was first recorded in U.K. by Stratford in 1839. FMD virus
was the first animal virus reported by Loeffler and Frosch in 1897 responsible for causing
the disease. The OIE (Office International des Epizootics) founded in Paris in 1924 was
the first international organization to become involved in the problem of FMD. IN India,
the work on FMD was started in 1943, under an ICAR -adhoc research scheme, at IVRI,
Mukteshwar. In 1971, the all India co-ordinated research project ( AICRP) for
epidemiological studies on FMD was initiated and it’s still continuing.

Distribution
FMD is endemic in Africa, Asia, South America and parts of Europe.The
disease can occur in any country but Japan, Newzealand and Australia are disease free.
In India the incidence is very high.

Properties
Virion properties are similar to family. During replication, for entry the FMD
viruses employ two receptors: initial binding involving heparan sulfate followed by high
affinity binding via integrins. FMD viruses can also enter cells via Fc receptors if virions
are complexed with non-neutralizing IgG molecules. This pathway, termed the antibody
dependant enhancement of infection pathway, is of unknown significance, but may be
important in the long-term carrier state that may occur in certain ruminants.
Host affected
Occurs naturally in cloven-hoofed animals (Both domestic and wild animals).
Cattle is most susceptible followed by pigs and rare in sheep and goats. Several wildlife
species including African buffalo, elephants, hedgehogs, deer and antelopes are also
susceptible. Man may contact the disease with mild symptoms such as vesicles on the
hand.

Morphology
One of the smallest animal viruses known. 18-25 nm in diameter. Spherical or
hexagonal in shape with 32 capsomeres. It has 2 distinct particles measuring 25 nm and 7
nm. The large part carries the infectivity property of the virus.

Physicochemical properties
Single cellular RNA, Mol. wt 2.8 x 10 6 daltons. Resistant to ether, chloroform,
bile salts and detergents. It is inactive at 56°C for 30 minutes. Stable at pH 7.4 - 7.6. Acid
labile (at pH 3 for 1-3 hours).
Resistance
The virus is stable at pH 7.4 - 7.6. Acid labile (at pH 3 for 1-3 hours). It is
inactive at 56°C for 30 minutes. Virus is readily destroyed when exposed to sunlight and
desiccation. But may survive for several weeks in tissue fragments or on contaminated
materials such as hay, straw, hair, hides etc

Virus in dried secretion and excretion on floors, walls bedding etc inside
contaminated buildings may be infective for atleast a month in summer and for 2 months
or longer in winter.

The virus in general is more resistant to many chemicals, which are very good for
other microbes. Phenolic disinfectants have little effect on the virus. So also alcohol,
ether, chloroform, acetone, bile salts, detergents and Tween 80 have no effect. Under
field condition 4% Na2Co3 (washing soda), 2% NaOH, 1% sodium meta silicate and 0.2%
citri acid are effective. In milk and milk products, the virus can survive at 70 0C for 15
seconds. But the virus in milk is destroyed at 70°C for 15 minute. 'The virus may survive
in sewage for several weeks. The haemagglutination property has not been demonstrated.

Antigenic property
7 main serotypes exist (O,A,C, SAT1, SAT2, SAT3 and Asia 1).
Seven serotypes of FMDV have been identified by cross –protection and
serological tests. They are designated as O (Oise), A (Allemaque), C (Carre), SAT1,
SAT2, SAT3 (South African Territories1,2 &3) and Asia 1.

Each virus type has been further subtyped on the basis of quantitative differences
in cross protection and serologic tests. Antigenic variation within a type occurs as a
continuous process of antigenic drift. So far 65 subtypes have bee identified. They are, O
= 11 subtypes, A = 32 subtypes, C = 5 subtypes, SAT I = 7 subtypes, SAT2 = 3 subtypes,
SAT3 = 4 subtypes, Asia 1 = 3 subtypes (65 subtypes).
The central laboratory at Mukteshwar, the antigenic charcterization was carried
out by microcomplement, micro neutralization, IPT, indirect and sandwich ELISA,
PAGE, PCR, monoclonal antibodies and immuno electrophoresis.
In our country only four major types, O, A, C and Asia 1 are known to occur.
Among, which A 22, A 5 & A 10 subtypes are more commonly occur.

Cultivation
Experimentally Guinea pigs, suckling mice, hamsters and rabbits can be infected.
Some strains grow in embryonated eggs (14 days old). Route is CAM or intravenous.

The virus grows very well in cell cultures. The cell cultures that are commonly
used are primary bovine tongue epithelium or bovine thyroid cells, primary pig kidney
calf kidney, and lamb kidney cells. Cell lines such as BHK-21 and IB-RS-2 cells are
highly suitable. Most strains of the virus multiply to produce CPE wthin 24-48 hrs. this
virus is highly cytolytic.

Epidemiology
In India, FMD most commonly occurs in Cattle, buffaloes, pigs, camel, sheep and
goats. Other domestic and wild animals, namely camel, elephant, mithun, deer, neelgai
bison and yak are also suffered from this disease. Man may contact the disease with mild
symptoms such as vesicles on the hand. Horse is refractory to FMDV infection.

FMD is also called as fast moving disease because

- highly infectious nature of the virus
- production of high titre virus in respiratory secretions
- large volumes of droplets and aerosols of virus shed by infected animals
- stability of virus in droplets
- rapid replication cycle with very high virus yield
- short incubation period
 The healthy animals are affected by the secretions and excretions of the FMD
affected animals. The excretion of virus for up to 24 hrs prior to onset of the symptoms
and the amount of virus ejected out by an infected animal helps in the spread of infection.
The titre of the virus during that period in milk or saliva is high about 10 5.5 infectious
units/ml. Virus is excreted in saliva, milk, urine and faeces after 2-8 days of infection.

Cattle, sheep and goats infected with FMD excrete virus and the amount varies
from 103 to more than 108 infectious units/day. Vesicular fluid contain high amount of
virus. Most of the air-borne particles are 6ug in diameter and normally fall on the ground
and spreads through winds. The aerosol spread is favoured by wind speed, low
temperature and high humidity. Spread occurs 60km or moreover land and 250 km over
sea.
In addition to inhalation, transmission also occurs by direct or indirect contact
with infected animals such as through abraded skin, conjuctiva, ingestion of
contaminated garbage, inoculation with contaminated vaccines and insemination.
Mechanical transmission may also occur with wild animals, birds and other non-
susceptible domestic animals.

Cattle, buffaloes, sheep, goats and humans act as carriers. The virus is persistent
in the pharyngeal region of cattle and buffaloe for up to 2 years or more and 9-11 moths
in sheep and goats. Virus may also persist in the mammary tisuue for 3-7 weeks. Virus
persistence does not occur in swine. Animals that have recovered from FMD are usually
resistance to infection with the same virus type for a year or more. But they can get
infected immediately with one of the other types or subtypes of FMDV.

In an outbreak of FMD, the roles of the primary hosts in transmission are as

follows:
 Sheep act as maintanence host
 Pigs act as amplifiers
 Cattle act as indicators
When sheep or goats become infected with FMDV, the disease may not be
diagnosed for a long time because signs and lesions are very mild. But during this time,
the animals will be producing infectious aerosols, contaminating fomites, and spreading
the virus by contact. FMD in pigs spreads more very rapidly, for they produce 30 to 100
times more virus in aerosols than sheep or cattle. (An infected pig can produce a hundred
million infectious doses per day). When cattle are infected with FMD, signs and lesions
usually develop more rapidly and are more severe than in pigs, sheep or goats. If cattle,
sheep, and pigs are exposed together, cattle usually get sick first.

Pathogenesis

The dose of infectious particles varies with the route of infection and
susceptibility of host. By respiratory route cattle require 10-100 infective virus particles,
while more than 105 virus particles are required by ingestion and only one infectious
virus particle is required through tongue. After entry in animal body the virus starts
multiplying, the common site is pharyngeal area and soft palate; in pigs it is bronchi,
bronchioles besides pharyngeal region. In sheep, it multiplies in tonsils, pharyngeal area
and soft palate.

Virus particles first attach to mucousal epithelial cells, penetrate into the
cytoplasm and replicate until the cells disintegrate. This release more viral particles to
infect other cells, including macrophages, which drain into the efferent lymphatic system
and then the blood. Viraemia results in further viral replication in lymph nodes,
mammary glands and other organs as well as the epithelial cells of the mouth, muzzle,
teats, interdigital space and coronary band.
Pathogenicity
Incubation period is 2 days to 3 weeks. In cattle, first indication is rise in
temperature which is more marked in young animals and associated with dullness, loss of
appetite and sudden drop in milk production. Within few hours vesicles gums and dental
pad and on the skin of the interdigital space, coronary bands of the feet, on teats and
udder. As the vesicles develop there is characteristic smacking of the lips, copious
salivation, and difficulty in chewing and marked lameness. The vesicles rupture in 2-3
days leaving raw, eroded red areas. Then the temperature drops and the affected animals
appear to gain some relief. Pregnant animals may abort.
Secondary bacterial infection occurs especially on feet. Myocardial damage is
severe in some outbreaks and is fatal in calves. Animals loose condition rapidly and
become emaciated. Morbidity is high and mortality is very low (less than 2%).
Symptoms in sheep, goats and pigs are similar to those of cattle except salivation
and obvious signs include sudden and acute lameness. The virus is present in secretions
and excretions during the viraemia stage (with in 7 days).

Clinical features

Incubation period is 2 days to 3 weeks. ( In pigs, the incubation period is as short

as 1-3 days)

Cattle
The onset is heralded by a precipitate fall in milk yield and a high fever ( 40-
0
41 C) accompanied by loss os appetite, depression. This is followed by appearance of
painful stomatitis and the temperature subsides. There is profuse salivation, the saliva
hanging in long, ropy strings, a characteristic smacking of the lips, and the animal chews
carefully. Vesicles and bullae (1-2 cm in diameter) appear on the buccal mucousa, dental
pad and tongue. The vesicles are thin walled contain a straw coloured fluid. Vesicles
rupture within 24hr, leaving a raw painful surface, which heals in about 1 week.

Vesicles appear on the feet, particularly in the clefts and on the coronet. The
lesions on the tongue often heal within a fewdays but those on the feet and nasal cavities
are contaminated with bacteria, maggot’s, which result in lameness and mucopurulent
discharge. Vesicles may also develop on the teats, which results in severe mastitis. The
virus does not cross the placenta but the abortion mostly during second trimester of
pregnancy mainly due to fever. Morbidity is high (reaches 100%) and mortality is very
low (less than 2%). Due to myocarditis the mortality, in young calves up to 6 months of
age, is very high.

A sequel to FMD in cattle, due probably to endocrine damage, is a chronic

syndrome of dyspnea, deleterious effects on testes causing production of poor quality of
semen, anemia, over growth of hair and lack of heat tolerance described as ‘panting’.

Sheep and goats

The pronounced clinical sign is sudden and acute lam eness. The vesicles develop
in the interdigital space of the feet, which rupture in about 2-3 days. Sometimes, the
upper layer of the hoof is lost. Oral lesions are rare or they may develop only on the
tongue and upper palate but there is no salivation.

Pigs
Large vesicles and bullae occur in the snout and feet and these may rupture to
expose large raw surface. Lamenesss is the first sign. The foot lesions are very painful.
Vesicles in the mouth are very less prominent.

Lesions
In tongue
 A small blanched whitish area develops in the epithelium
 Fluid fills the area, and a vesicle is formed
 Vesicle enlarges, coalesce and rupure leaving an eroded area
 Gray fbrinous coating which later becomes yellow, green or brown.
In feet addition to vesicles, separtion of the skin and horn and coronitis is seen
When the death takes place, the heart muscle shows degeneration and necrosis
leading to the formation of white streaks in heart muscles. (Tiger heart).

Ecology: The virus spreads by both direct and indirect means.

Direct
From animal to animal by droplet infection or when the virus contacts the mucosa
of the mouth, nose, conjunctiva, or abraded skin surface. Virus is excreted in saliva, milk,
urine and faeces after 2-8 days of infection.

Indirect
Spreads mostly through contamination of food, water, bedding and pasteurs by
the infected discharges of diseased animals. Airborne infection may also occur.
Migratory birds spread the disease.

Diagnosis
The highly contagious nature of the disease, profuse salivation and the presence
of typical raised vesicles with blanched covering epithelium filled with a clear straw
colour fluid is usually pathognomonic.

Samples to be collected
 The specimen usually collected is vesicular fluids, epithelial fragments of recent
vesicles especially from lesions of tongue, feet, udder or lips of about 1 gm. In recently
dead animals pieces of cardiac muscle and pancreas should be collected. The samples
should be kept in a solution of phosphate buffer saline at pH 7.6 which contain equal
parts of glycerol and 0.00196 phenol red indicator. The outside of the container should be
disinfected with 4% Na2 Co3 or 0.2% citric acid. The container is wrapped with cotton
wool and kept in a sealed fluid tight container packed in a strong outer box of wood and
sent to the lab.

By Serological tests
Complement fixation test (CFT) is the most commonly used and important test to
identify the type of virus. The suspension of the original serum is used as antibody.
Virus and serum neutralization tests may be used to detect specific antibodies in
the serum of recovered animals or for identifying the causative virus. The test is done
with known hyperimmune sera. Gel diffusion test, FAT, IPT and Iso electric focusing are
also useful.
ELISA and PCR are now standard methods for virus detecion and typing in
reference laboratories.
Cell culture - Monolayers of bovine thyroid or pig kidney cells are inoculated and CPE
develops with in 24-48 hours at 37°C.

Mouse inoculation
The suspected material is inoculated in unweaned mice and the mice dies within
1-7 days.

Cross immunity
For accurate identification inoculation of groups of susceptible and immune cattle
are used.

Differentail diagnosis
Animal species Route of inoculation FMD VS VE SVD
Natural infection
Cattle + + - -
Pig + + + +
Sheep and Goat + + - -
Horse - + - -
Experimental infection
Horse Intra dermal in tongue - + + -
Guinea-pig Intra dermal in foot pad + + - -
Suckling mice Intra dermal in foot pad + + - +
Adult chicken Intra dermal in tongue + + - -
VS- Vesicular stomatitis, VE- Vesicular exanthema, SVD – Swine vesicular disease

Control
 Spread by air borne route is very difficult to control. Control of movement of
livestock is one of the effective measures. Where the disease is not endemic the policy of
quarantine, slaughter and disinfection of infected premises has proved efficient and
economical also.
Where the disease is not endemic the policy of quarantine, slaughter and
disinfection of infected premises has proved efficient and economical also.
In endemic areas aphthization is done (ie. immunization by deliberate spread of
the disease). Cattle in the infected areas are inoculated with live virus and held in
quarantine for a few weeks while the disease is spreading through the group. This method
is dispensed with because it may aid in the spread of the disease. Virus inactivated with
formation and adsorbed on to alum may be used. Previously vaccine was prepared by
tongue epithelium of infected cattle. Recently the virus is grown in cell cultures of bovine
tongue epithelium or in BHK21. A lapinised virus vaccine is also used (Rabbit adapted).
Vesicles containing live virus is attenuated by passage in adult mice, cell cultures or eggs.
FMD virus vaccines are produced as monovalent preparations and mixed before use to
get polyvalent vaccines.
In India, there is no vaccine available for pigs. For the control of disease,
programmed application of vaccines in cattle, buffaloes, sheep and goats have to be
followed to bring down the disease outbreaks. A practical approach in this direction has
been demonstrated by NDDB by launching FMD control programme in Nilgiri district of
Tamil Nadu in 1983. The Indian dairy development corporation has exended the
programme in 13 districts of Tamil Nadu, 5 districts in Karnataka and 9 districts of
kerala. Under this, all the cattle, sheep and goats are vaccinated at 6 monthly intervals for
one and a half year, followed by ring vaccination in 15 km belt and a 3km belt on either
side of cattle routes. An information system reporting of FMD outbreak has been
developed. An epi-demiological surveillance of the project area is carried out. The strains
of virus causing outbreak have been studied in detail through AICRP on FMD.

Note: when vaccinating animals, it is important that the vaccine contain the same subtype
of virus as is in the area. This necessitates frequent checking of the serotype and subtype
during an outbreak because FMD virus frequently changes during natural passage
through various species.

Avian Enteroviruses
AVIAN ENCEPAHLOMYELITIS

Synonym: Epidemic Tremor

Acute viral disease of very young chickens (1-21 days) characterized by ataxia,
paralysis, tremors ih head and neck. AEV is not pathogenic to older chicken.

This diease was first described in the New England states of the U.S.A. in 1932
and is now recognized worldwide. In addition to chickes this disease has also been
recorded in pheasants, quail and turkeys.

Properties
 Avian encephalomyelitis caused by avian entero virus belongs to the genus
picornaviridae. Morphology is similar to the family. This virus is reisitant to lipid
solvent, acid pH, trypsin and pepsin. The virus is able to survive very well at ambient
temperature. Formaldehyde fumigation is effectively inactivating AEV with an 18hr
exposure period. Only a single antigenic type but strains vary in virulence. AEV grows
very well in yolk sac route of embryonated chicken eggs, chicken embryo fibroblast and
chicken embryo kidney cell culture.

Transmission

AE is an enteric infection and affected chicks shed the virus in the faeces for
about 2 weeks. The virus is live in litter material for longer time. Horizontal spread by
fomites occurs. Vertical transmission is a very important mode of transmission. Breeder
hens get infected. A proportion of the eggs of infected hens are infected. Chicks infected
in ovo hatch normally but shed virus in their droppings and infect other chicks in the
incubator shortly after hatching.

Pathogeneis
Egg adapted strains are highly virulent and neurotropic. Natural strains are
enterogenic.
Oral route

Alimentray canal

Vireamia

Infection of CNS (Purkinje cells of the cerebellum)

Variety of visceral organs and tissues (Pancreas, Heart, Kidney, Liver & Spleen)

Replication of the virus in intestinal tract followed by oral route results in

viraemia and a competent immune response is required to prevent infection of the central
nervous system. Affected chicks excrete the virus for 2 weeks after second week of
exposure.

Clinical signs
Incubation period is 1-7 days. Affected chicks exhibit dullness, depression,
progressive ataxia, tremors in head & neck and weight loss. They fall on one side or sit
back on their hocks or refuse to move. Blindness, paralysis, prostration and coma also
occur. Death is mainly due to affected chicks nto able to take feed and get trampling by
penmates.
Lesions
No characteristic lesion. Nonsuppurative encephalomyelitis, which is,
characterized by perivascular cuffing, neuraonl degeneration and microgliosis. Large
amount of lymphocytic accumulations in viscera especially in pancreas, myocardium and
proventriculus are characteristic. Mononuclear cell infiltration in the iris leads to cataract.

Diagnosis
Samples to be collected
Brain is the best choice. Pancreas also suitable. Inoculate suspected material into
yolk sac route of chicken embryo. Allow the egg to hatch out and observe characteristic
signs in chick for 7 days.

Demonstrate the viral antigen in impression smear by FAT & IPT taken from
brain.

Embryo susceptibility test

Inoculate embryo-adapted strain into yolk sac route in embryonated eggs. At the
age of 18 day old, all the (100%) embryos exhibit paralysis and severe muscular
dystrophy.
Diiferential diganosis should be made with New Castle diseas

Control
1. Depopulation
2. Attenuated live-virus vaccine administered in the drinking water to be
administered after chickens reach 10 wks of age or 4 weeks before laying--
Passive immunization. High levels of maternal antibodies protect the chicks up to
21 days.
3. Inactivated vaccine.

DUCK VIRAL HEPATITIS

Acute, highly contagious, usually fatal disase of very young ducklings (less than
21 days of age) characterised by sudden onset, rapid spread and high mortality.

Duck viral hepatitis was first recognized in long Island, New York, USA in 1945.
In India this disease was first reported by Rao and Gupta in 1967.

Properties
Duck viral hepatitis caused by avian entero virus belongs to the genus
picornaviridae. Morphology is similar to the family. This virus is reisitant to lipid
solvents, acid pH, trypsin and pepsin. It is stable 50 0C for one hour. Inactivated at 62 0C
for 30 mts. Three sero types are there (DHV-1, DHV-1& DHV-3). Among which DHV –
1 is highly prevlent in India. DHV grows very well in allantoic route of duck, Goose and
chicken embryonated eggs. Dead occur within 4 days post inoculation. Stunted growth,
odema, patchy haemorrahges on embryo, enlarged and greenish liver and greenish
discouration of the allantoic fluid are characteristic.
Cell cultures of duck origin, such as, duck embryo liver cells, duck embryo
kidney or, duck kidney are highly suitable.

Transmission
Domestic ducks are natural host. Turkey, quail and guinea fowl are also
susceptible. DHV is an enteric infection and affected chicks excrete the virus in the
faeces upto 8 weeks. Because of the hardy nature of the virus the horizontal spread (fecal
–oral route) transmission occurs very rapidly.

Pathogenesis
Oral route is the portl of entry. Pathogenesis not well characterized. The virus was
isolated from various visceral organs, blood and faeces.

Clinical signs
Incubation period is very short i.e. less than 24 hrs only. Disease occurs most
commonly in ducklings less than 21 days of age. Affected ducklings isolated from the
penmates, stand still with partially closed eyes, fall to one side, kicked spasmodically and
displayed opisthotonus at death (is very characteristic). The interval between the first sign
and death was not more than an hour. Within 3 days period mortality reaches 100%.

Lesions
Red, soft, enlarged liver (twice the normal), oedematous, mottled with echymotic
haemorrhages. Spleen and kidney are enlarged. Histologically extensive hepatic necrosis,
inflammatory cell infiltration in various organs, proliferation of bile duct epithelium,
encephalitis, perivascular cuffing and gliosis are seen.

Diagnosis
i) History, Clinical signs, post mortem examination.
ii) Demonstration of viral antigenin various tissues by lmmunofluorescence.
iii) Young ducklings (1 to 7 days) are most suitable for virus isolation. Virus
isolation in cell culture or by allantoic inoculation of 10-day-old embryonated hen's eggs.
Infected eggs when candled often show characteristic greenish discolouration of the
embryonic fluid. Most are dead within 4 days after inoculation. Differential diagnosis
should be made with ND, Duck Plaque and influenza.

Control
Recovered are ducks are immune. Hyperimmune serum is useful to reduce losses
during outbreaks. Attenuated egg-adapted live virus vaccines are useful to vaccnate
breeder ducks.

Enterovirus
Teschen and Teschen-like Diseases
(Porcine encephalomyelitis)
Cause
The enteroviruses causing these diseases have been placed in one serological group,
which is further divided into three subgroups. One of these subgroups includes porcine
teschovirus 1; originally called porcine enterovirus serotype 1, the cause of Teschen
disease. The cause of Talfan disease is in another subgroup.
Occurrence
Teschen disease, which is a severe porcine encephalomyelitis, is thought to occur rarely
only in Europe and Africa. The less severe form, Talfan disease, probably occurs
worldwide.
Transmission
The virus is shed in feces and saliva; infection is by ingestion. Spread is by direct and
indirect contact.
Clinical & Pathologic Features
These viruses usually cause an infrequent disease in young pigs. The incubation period is
approximately 10 days and pigs of all ages are affected during outbreaks. The most
serious outbreaks (Teschen) have been reported from Europe.
Clinical signs are fever (104 to 106°F), ataxia, stiffness, tremors, dog-sitting position,
nystagmus, convulsions, and prostration. Deaths usually occur within a few days after
onset of signs. Mortality in Teschen disease is 50 - 75% or higher in young pigs.
The lesions include glial nodules, neuronal degeneration and lymphocytic perivascular
cuffing mainly in the spinal cord.
Talfan disease is clinically similar to Teschen disease but is less severe and the fatality
rate is lower. The disease may be confused with pseudorabies, hog cholera, and
hemagglutinating encephalomyelitis virus (HEV) infection.
Diagnosis
 Clinical specimens: Brain, spinal cord, and intestine.
 These viruses can be propagated in cell cultures of swine origin in which they
produce cytopathic changes. Identification is accomplished by virus neutralization
tests using specific antisera. With regard to significance of viral isolation, it
should be kept in mind that enteroviruses occur commonly in the intestine.
Prevention
 Animals that recover are immune.
 Live modified virus and an inactivated virus vaccine have been employed with
success in Europe.

Swine Vesicular Disease

Cause
Swine vesicular disease virus (SVDV) of which there are several antigenically different
strains. The SVD virus is closely related to the human enterovirus, Coxsackie B-5.
Occurrence
This disease was first reported in feeder pigs in Italy in 1966. It has since been reported in
Great Britain (declared free in 1980), several European countries, and Asia. There have
been reports of aseptic meningitis in humans caused by SVDV.
Transmission
Spread is by direct contact and fomites. Several outbreaks have been traced to uncooked
or insufficiently cooked pork in garbage.
Clinical & Pathologic Features
The disease is clinically indistinguishable from foot-and-mouth disease (FMD), vesicular
stomatitis (VS), and vesicular exanthema (VE).
Epithelial tissue is initially involved, followed by viremia with generalized infection of
lymphoid tissues.
The first signs are reduced feed intake, lameness and tenderness of the feet, fever up to
106°F, and the formation of vesicles on the feet, snout, tongue, mouth, nostrils, and teats.
The prognosis is favorable, but in most countries infected animals are slaughtered.
Experimentally infected pigs may show central nervous system involvement.
Diagnosis
 Clinical specimens: Vesicular fluid, affected skin and mucous membranes, blood
with anticoagulant, and serum.
 The virus can be propagated in cell cultures of porcine kidney. Cytopathic
changes typical of picornavirus are produced in 2 - 4 days.
 In many countries regulatory officials should be contacted if SVD is suspected.
They will collect appropriate clinical specimens to be tested, typically at a central
government laboratory.
 Among the tests used is an ELISA to detect viral antigen in vesicular material.
Because SVD resembles clinically FMD a correct diagnosis is imperative.
Prevention
 Vaccines are not available to prevent SVD.
 SVD is a reportable disease and, if it is suspected, strict measures of control
including quarantine and slaughter are implemented.

Porcine Enteroviruses: Reproductive Infections

(SMEDI virus syndrome)
Cause
Porcine enteroviruses (Enterovirus). The viruses are designated by the name SMEDI,
which is derived from Stillbirth, Mummification, Embryonic Death, Infertility).
Occurrence
The causal enteroviruses occur widely in swine herds.
Transmission
The viruses are present in feces and infection is presumably by direct and indirect
contact.
Clinical & Pathologic Features
Susceptible pregnant gilts or sows are infected initially and then their embryos or fetuses.
The SMEDI viruses have been isolated from stillborn pigs, "3-day" dead pigs, and fetuses
found dead in the uterus in mid-pregnancy after hysterectomy. Herds from which these
enteroviruses were recovered generally had small litter sizes and low survival rates.
Sows were immune after infection but the disease manifestations frequently appeared
cyclically every 2 - 3 years; perhaps because of the susceptibility of new gilts. The typical
disease manifestations of this group of viruses were produced experimentally. The five
SMEDI virus groups reported fell into four groups of serologically related viruses. At
least six other enterovirus groups have been identified that are not associated with
reproductive problems in sows.
Porcine parvovirus is considered to be the primary viral cause of reproductive problems
in swine and little significance is given to the SMEDI viruses.
Diagnosis
 Clinical specimens: Tissue from fetuses and stillborn pigs.
 The viruses can be cultivated in primary pig kidney cell cultures; however, the
frequency of isolations has been low, mainly because of the absence of viable
virus at the time clinical disease becomes apparent.
Prevention
There are usually no attempts to prevent or control enterovirus infections in swine.

Bovine Enteroviruses
Two serotypes are recognized. Serotype 1 has been isolated from a wide range of animals
species, while serotype 2 has only been recovered from cattle. Most infections are
subclinical but abortion, stillbirths, and infertility in bulls has been reported.

Avian Enteroviruses
These include: duck hepatitis virus 1, duck hepatitis virus 3, avian nephritis virus and
enteroviruses recovered from normal birds. Some strains have the capacity to cause
serious hepatitis and nephritis in chickens and turkeys.

Caliciviridae
This family is composed of small, non-enveloped, positive-sense single-stranded RNA
viruses with distinctive morphological features. The viruses infect a wide variety of
animals and two are important pathogens of domestic animals.
Viral Characteristics
 Non-enveloped, positive sense, ssRNA viruses, 35 - 40 nm in diameter with
icosahedral symmetry
 The 32 cup-shaped depressions on the spherical capsid surface give caliciviruses a
distinctive morphology.
 The genome is a single molecule of linear single-stranded RNA (7400 to 8300
nucleotides in length) with positive polarity. It contains a 5' protein (VPg) or a
methylated nucleotide (for hepatitis E) and a 3' polyA tail.
 The genome alone is infectious.
 Replication takes place in the cytoplasm and virions are released with cell lysis.
 The virions are stable in the environment and resistant to some disinfectants;
sodium hypochlorite is effective.
Classification
The Caliciviridae consists of four genera, Vesivirus, Lagovirus, "Norwalk-like viruses"
and "Supporo-like viruses". The last two categories contain viruses that infect humans.
The first two categories contain viruses of veterinary significance as follows:
 Vesivirus
o Vesicular exanthema virus
o Feline calicivirus
o Canine calicivirus: Not considered of pathogenic significance; recovered
from dogs with diarrhea.
 Lagovirus
 o Rabbit hemorrhagic disease virus
o European brown hare syndrome virus
 "Norwalk-like viruses"
o Norwalk virus: A major cause of acute gastroenteritis in humans.
 "Sapporo-like viruses"
o Many non-pathogenic strains recovered from humans.
Vesivirus
Vesicular Exanthema
(San Miguel sea lion virus disease)
Cause
Vesicular exanthema of swine (VES) virus. Thirteen serotypes of the virus designated A,
B, C, etc, have been identified to date. A number of serotypes have been implicated in
San Miguel sea lion disease.
Occurrence
The virus naturally infects swine; horses and dogs. Hamsters can be infected
experimentally.
The disease, with the exception of isolated outbreaks in Hawaii and Iceland, has only
been seen in the continental United States. It has not been reported since 1956.
A virus closely related or identical to VES virus was isolated from San Miguel sea lions
along the California coast. It was capable of producing vesicular lesions in pigs
experimentally inoculated. Several serotypes of this virus affect other marine mammals,
including northern fur seals, and antibodies to these viruses have been detected in a
variety of terrestrial mammals. It has been suggested that marine animals are the reservoir
of VES virus.
Transmission
Vesicular exanthema virus is transmitted by direct and indirect contact and by fomites.
The virus is present in the saliva and feces of infected pigs. It has been conjectured that
pigs may have been infected by eating food containing portions of sea lion carcasses.
Clinical & Pathologic Features
The disease in swine clinically resembles foot-and-mouth disease but is milder. There is
fever and vesicles appear on the snout, the mucous membranes of the mouth, the feet, and
the udder of nursing sows about 48 hours after viremia. The vesicles break in 24 - 48
hours leaving erosions that rapidly are covered with new epithelium. The disease is
usually benign and runs a course of about two weeks.
Diagnosis
VES is an important disease because of its similarity to foot-and-mouth disease, vesicular
stomatitis and swine vesicular disease.
 Clinical specimens: Vesicular fluid, affected mucous membranes, blood with
anticoagulant, and serum.
 Vesicular fluid contains virus that can be identified by complement fixation and
ELISA and detected by electron microscopy.
 The virus can be cultivated in swine embryonic kidney cells in which cytopathic
changes are observed. In cell cultures, the virus can be identified by neutralization
tests and viral antigens can be detected by immunofluorescence.
Prevention
 The disease is now considered eradicated from the United States. The last
occurrence was in 1956. However, the possibility of infection from marine
mammals must be kept in mind.
 It is a reportable disease and is dealt with by the same strict measures used to
control other vesicular diseases.

Feline Calicivirus Infection

Cause
Feline calicivirus of which there are several serotypes.
Occurrence
Feline calicivirus infection is highly contagious, occurs worldwide, and rivals feline viral
rhinotracheitis (FVR) in frequency of occurrence.
Transmission
Spread is by direct and indirect contact and the mode of infection is mainly by inhalation
of aerosols.
Clinical & Pathologic Features
Infection begins in the oropharynx with extension to the upper respiratory tract and eye.
The incubation period (2 - 5 days), course, and clinical manifestations resemble FVR.
Clinical signs include fever, mild rhinitis, sneezing, nasal discharge, conjunctivitis,
palatine or glossal ulcerations, and, in some instances bronchopneumonia. There is a
great variation in the severity of the disease, probably owing to variability in the
virulence of different strains of the virus and host factors. Kittens 2 - 6 months of age are
most severely affected.
The fatality rate can be substantial in kittens and older cats. Infected cats become
carriers and the virus is shed continuously from the pharynx and tonsils for months and
sometimes for years.
Abortion may occur in queens.
Diagnosis
 Clinical specimens: Nasal, oropharyngeal, and ocular swabs; lung and trachea
from necropsied cats.
 Isolation and identification of the virus are required for a definitive diagnosis. The
virus can be easily propagated in cell cultures of feline origin, in which a rapid
and well evidenced cytopathic effect is produced. Upon production of
characteristic CPE, the virus can be identified by virus neutralization with specific
antiserum.
Prevention
Feline calicivirus is strongly immunogenic, and killed and modified-live vaccines of cell
culture origin are available, often in combination with other viruses. They are
administered intranasally or intramuscularly.
Lagovirus
Rabbit Hemorrhagic Disease
Cause
Rabbit hemorrhagic disease virus.
Occurrence
Rabbit hemorrhagic disease (RHD) affects domestic and wild rabbits and hares.
Destructive outbreaks have been reported worldwide. Although not endemic in the USA
outbreaks have occurred, most recently in 2001 and 2005.
Transmission
Spread by is by direct and indirect contact and mechanically by mosquitoes and other
insects. The virus is present in secretions and excretions.
Clinical & Pathologic Features
Infection is by the oral/fecal route and the principal target cells are mononuclear
phagocytes. The disease is highly contagious and infections are characterized by acute
onset and frequently death without premonitory clinical signs. Some affected rabbits may
have signs of respiratory distress, such as dyspnea, abdominal respiration, and mild
nosebleed. Swollen, congested and hemorrhagic viscera are noted at necropsy.
Histologically, the most characteristic lesion is a coagulative hepatic necrosis. There are
hemorrhages in the lung and focal areas of necrosis in the spleen and heart. Severe crypt
necrosis may be ocasionally noted in the small intestine.
Diagnosis
 Clinical specimens: Liver, spleen and lung.
 A presumptive diagnosis is made on the basis of the peracute nature of the
disease, liver necrosis and other characteristic gross lesions.
 Virus in tissues can be detected by electron microscopy and antigen identified by
ELISA and immunofluorescence.
 The virus in tissue preparations will agglutinate human type O erythrocytes.
 The virus has not been propagated in cell cultures.
Prevention
 Where the disease is endemic inactivated vaccines are used.
 Outbreaks in North America are dealt with by strict quarantine and slaughter.
 The disease is of such severity that it has been used to reduce rabbit populations.
European Brown Hare Syndrome
The disease caused by this lagovirus is similar to rabbit hemorrhagic disease (RHD) and
has been reported in wild and domestic hares in many European countries. The European
brown hare syndrome virus is antigenically similar to the virus that causes RHD.
This hemorrhagic disease with liver necrosis has a high mortality rate in adult hares.
Astroviridae
This is a family of small, single-stranded RNA viruses that are recovered from a wide
variety of birds and mammals and are rarely associated with clinical disease. The name of
the family comes from the appearance of viral particles under EM examination; some
Astrovirus virions display a five or six-point star-like appearance.
Members of the Astroviridae are species-specific and included in a single genus,
Astrovirus. Different antigenic types are recognized; specifically three types have been
recovered from cattle and seven from humans.
Viral Characteristics
 The genome consists of a linear, positive-sense single-stranded RNA molecule.
 They are naked viruses with icosahedral symmetry (27 - 30 nm in diameter).
 Some virions possess a star-like appearance with five or six-point surface
projections.
  Replication takes place in the cytoplasm and viruses are released by cell lysis.
 They have considerable resistance to heat and detergents.
Significance
Astroviruses were initially observed by electron microscopy examination of children’s
feces and were subsequently observed in feces of dogs, cats, sheep, cattle, swine, ducks,
turkeys and other animals.
Mild gastroenteritis due to astroviruses has been reported in cattle and other
domestic species. The disease associated with astroviruses is ordinarily self-limiting.
Mild gastroenteritis has been reported in avian species. Young ducks may develop an
acute fatal hepatitis.
Astroviruses have also been implicated as a cause of a watery diarrhea in children.
Astrovirus gastroenteritis is uncommon in adults; however, many adults have antibodies
attesting to the ubiquity of the virus.
Laboratory diagnosis is based on observation of large numbers of astroviruses in
feces by electron microscopy.

Togaviridae

This is a family of enveloped, positive sense, single-stranded linear RNA viruses. Only
one of the two genera, Alphavirus, has viruses of veterinary significance. They are
arboviruses causing important equine encephalidides.
Viral Characteristics
 They are enveloped viruses (~ 70 nm) with an icosahedral nucleocapsid
containing a single linear, positive sense, single-stranded RNA.
 The genome has a 5' cap and a 3' poly A tail.
 The envelope has characteristic glycoprotein spikes.
 They replicate in the cytoplasm and the nucleic acid alone is infectious.
 The virions mature by budding from the plasma membrane.
 They agglutinate goose and chick erythrocytes.
 They are labile in the environment.

Classification
The family Togaviridae has two genera, Alphavirus and Rubivirus. Only Alphavirus
contains viruses of veterinary significance. The diseases caused by the viruses of each
genus are as follows:
 Alphavirus: The following are significant veterinary alphaviruses:
o Eastern Equine Encephalomyelitis virus
o Western Equine Encephalomyelitis virus
o Venezuelan Equine Encephalomyelitis virus
o Highlands J virus. This virus was originally thought to be a WEE virus
that occurred in the eastern USA. It has been isolated from rodents, wild
and domestic birds, and mosquitoes in the eastern USA and is now
considered a distinct alphavirus.
o Getah Virus
 Rubivirus
 o Rubella virus: The cause of rubella (German measles) and the more
serious congenital rubella syndrome involving serious human fetal
abnormalities.
Alphavirus
Eastern Equine Encephalomyelitis (EEE),
Western Equine Encephalomyelitis (WEE) and
Venezuelan Equine Encephalomyelitis (VEE).
General
Cause
Each of the three equine encephalidides, EEE, WEE and VEE is caused by a
serologically distinct Alphavirus designated as WEE virus, EEE virus and VEE virus.
RNA sequence analysis shows that most of the WEE virus genome is closely related to
that of the EEE virus genome. The constituents of these "categories" are referred to
below.
Transmission / Reservoir
Mosquitoes of various genera are the principal vectors. These viruses infect and replicate
in mosquitoes for their entire life. In tropical and subtropical regions the viruses are
endemic in swamp habitats in a bird/rodent-mosquito cycle. It is not clear how the viruses
are maintained from one year to another in temperate regions.
Pathogenesis
After initial infection the virus travels to lymph nodes via the lymphatics. It replicates in
neutrophils and macrophages followed by viremia and replication in other organs,
including the brain. Neurologic signs, when seen, develop in less than a week of
infection. Subclinical infections occur with all of these viruses.
Clinical Signs
Infections are most common during summer and early fall when mosquito populations
are high. After exposure, there is an incubation period of about 1 to 7 days followed by
fever, depression, anorexia, sopor, pharyngeal paralysis, head pressing, incoordination,
paralysis of the legs, recumbency and death frequently occurs in 2 - 7 days.
Specific
Eastern Equine Encephalomyelitis
 EEE is caused by two antigenic variants, the North American and the South
American.
 Causes disease in equids, pigeons, pheasants, quail and humans.
 The North American variant occurs in the Caribbean, states east of the
Mississippi, Texas and eastern Canada. The South American variant occurs in
Central and South America. It is less pathogenic than the North American variant.
 The principal vectors are mosquitoes. In North America, EEE is commonly
transmitted by Culiseta melanura and other mosquito species in some regions.
 The reservoir hosts are mainly wild birds and small rodents.
 It is estimated that ~10% of the horses infected develop clinical disease.
 The mortality in horses may reach 90%; in humans 30 - 50%.

Western Equine Encephalomyelitis

 The genome of WEE virus is closely related to that of EEE virus. Several
subtypes have been identified including Sindbis, Fort Morgan, and Aura, which
 are not important causes of equine encephalitis. It has been suggested that WEE
virus arose as recombinant between Sindbis virus and EEE virus.
 Causes disease in equids and humans.
 Occurs in western Canada and in states west of the Mississippi in the USA,
Mexico and South America.
 Principal vectors are Culex and Aedes spp. mosquitoes and the tick Dermacentor
andersoni.
 The reservoir hosts are wild birds; the avian and mosquito infections are harmless.
 WEE is milder than EEE. The mortality in horses is 20 - 30%; in humans about
10%.

Venezuelan Equine Encephalomyelitis

 Six antigenically related subtypes of VEE virus have been identified and given
names. The most important subtype is designated subtype 1; it contains five
serovars of which several are the principal causes of VEE.
 Causes disease in equids and humans.
 Occurs in the Central and South America, Mexico and infrequently in the
southern USA Subtypes other than 1 have the same distribution and are enzootic
with a rodent/bird-mosquito cycle. They are not causes of VEE.
 The principal vectors are considered to be mosquitos and hematophagous insects.
 The reservoir hosts are birds and small forest rodents.
 The virus of VEE is shed in oral secretions and contact transmission may occur.
 The disease is more viscerotropic than neurotropic with damage to blood vessels
and lesions in many organs including less frequently the brain.
 The mortality rate in horses may approach 80%; in humans, who suffer a milder
disease, mortality is about 1%.
General
Diagnosis
 Clinical specimens: Whole blood collected during the febrile stage and brain
tissue from horses that have died. Acute and convalescent sera.
 A presumptive diagnosis is often based on clinical signs and microscopic brain
lesions, which consist of an inflammatory cell infiltrate with perivascular cuffing,
and congestion and edema of the meninges (EEE and WEE). A neutrophilic
inflammatory cell response is characteristic of EEE virus infection.
 Confirmation requires isolation and identification of the virus. The viruses can be
propagated in various cell cultures and in young mice inoculated intracerebrally.
 A definitive diagnosis can also be obtained by demonstrating a significant
increase in specific antibody between acute and convalescent sera. The
procedures used include hemagglutination-inhibition, virus neutralization,
complement fixation and capture ELISA.
 A presumptive diagnosis can be made on the basis of signs and a single positive
serum sample, if the horse has not been vaccinated.
Treatment
 Steroids to reduce inflammation.
 General supportive care.
  Horses that recover and have had neurologic signs frequently have neurologic
deficits.

Getah Virus Infection

An outbreak of Getah virus infection occurred in racehorses in Japan in 1978. The
infection was mild and characterized by fever, edema of the hind legs, and urticaria.
Recovery was uneventful in about seven days.
Getah virus has been isolated from mosquitoes in Japan, Southeast Asia, and Australia.
Serologic studies indicate that infections occur in a number of other animal species,
including cattle and especially pigs. The virus has been incriminated as a cause of fetal
death in infected sows. Transmission was thought to be by mosquitoes principally.
The virus can be propagated in cell cultures and in mice inoculated intracerebrally.

Family :Flaviviridae

Members of the family are 40 to 60 nm in diameter with icosahedral capsids and

tightly adherent envelopes. The genome is composed of positive sense single stranded
RNA. 5' end capped, but 3' end usually not polyadenylated, infectious: genes for
structural proteins located at 5’ end of genome. Cytoplasmic replication; polyproteins
translated from genomic RNA are cotranslationally cleaved to yield several nonstructural
proteins and three or four structural proteins: maturation occurs on intracytoplasmic
membranes without evidence of budding and release by exocytosis.
The family comprises three genera namely Flavivirus, Pestivirus and Hepacivirus.
Two genera, Flavivirus and Pestivirus contain viruses of veterinary importance. Most
members of the genus are arboviruses, which require either mosquitoes or ticks as
vectors. Viruses in the genus agglutinate goose red cells. The genus Pestivirus contains
three viruses of veterinary importance namely bovine viral diarrhoea virus, border disease
virus and classical swine fever virus.

PESTIVIRUS
SWINE FEVER
Synonym :Hog cholera

Highly contagious viral disease of pig characterized in acute cases by sudden

onset, marked haemorrhage of internal organs and high mortality.

Host
Swine. The only animal in which the disease occurs naturally.

History and distribution

First described in Ohio in 1833. In 1903 Salmonella choleraesuis was found to be
the secondary invader. Outbreaks were recorded in England, France, Germany, Spain,
Africa, India, Australia, and Japan.

Morphology
The virions are small spherical particles measuring about 40 nm in diameter.
Physiochemical properties
It is sensitive to ether and other lipid solvents. Moderately sensitive to Trypsin. It
is a stable virus and virulent in blood remains viable for a considerable period when
stored at +4°C. Virus in defibrinated blood is more stable when held at pH 4.8-8.5 than at
neutral pH, very sensitive to putrefaction and looses infectivity with in 3-4 days if carcass
is allowed to decompose. In the other hand virus persists for long periods in infected pork
and garbage and also in meat kept in cold storage. The virus is fairly resistant to most of
the ordinary disinfectant. 5% NaOH destroy the virus in one hour.

Antigenic properties
Only one serological type has been recorded. Recovered pigs are permanently
immune.

Cultivation
Cultivation is done originally in susceptible pigs. Later on various cell culture
systems derived from swine tissues have been used for primary isolation and for
fundamental studies of the virus. The tissue is most commonly used for cell cultures
including kidney, spleen, bone marrow, testis, lymph node and leucocytes. There is
usually no evidence of CPE or only minimal. In 1958 it was observed that a mild CPE of
Newcastle disease on monolayer culture is enhanced by all strains of swine fever virus.
The marked cellular degeneration that results is a reliable method for detecting the
presence of swine fever virus in suspected tissues. Swine fever immune serum
specifically inhibits the exalting effects of swine fever virus on Newcastle virus. This test
is called as exaltation of Newcastle disease or END test. Virus passage directly through
rabbits or by alternate passage with the pig and rabbit is modified and looses its virulence
so that it can be used as a live attenuated virus for pigs.

Epidemiology
Infection is transmitted readily by direct and indirect contact. The virus is usually
acquired by ingestion of food and water contaminated with discharges and secretions
from infected pig. Urine and nasal and ocular discharges are generally regarded as most
infective.

Pathogenesis
Pigs are usually infected by the oronasal route. The tonsil is the primary site of
viral multiplication. Virus spreads to regional lymph nodes and viraemia develops after
further viral multiplication. Virus has an affinity for vascular endothelium and
reticuloendothelial cells can be isolated from all major organs and tissues.

Pathogenicity
Domestic pigs of all breeds and ages especially young animals are highly
susceptible. Incubation period is 3-8 days. The earlier signs are dullness, restlessness,
anorexia accompanied by marked thermal response (41-42°C). Most affected animals
show apparent weakness in hindquarters manifested by in coordination of movement and
staggering gait. In many cases there is conjunctivitis, vomiting, constipation followed by
diarrhoea. Red or purple haemorrhagic area may appear on the skin of abdomen or on the
ears and snout and there may be raised cutaneous necrotic areas. Course of the disease
varies, usually it is 5-16 days. Although in extremely acute cases the number of pigs
found dead in a overnight without having shown any signs. Mortality extremely high over
90%. High mortality usually associated with secondary bacterial infection of intestine and
lungs. The virus enters the body either by ingestion or by inhalation. The virus has got
special affinity for tissues of mesodermal origin particularly haemopoietic and vascular
tissues. Damage to these cells leads to enlargement of lymph nodes and wide spread
haemorrhages.
Postmortem lesions include degeneration of small blood vessels leading to
haemorrhages in kidney, bladder, skin and lymph nodes. Circular or oval raised button
are most prevalent in caecum and proximal portion of the colon. Turkey egg appearance
of kidney is the pathognomonic lesion of the disease.

Diagnosis
In typical cases tentative diagnosis is based on characteristic symptom and
lesions.
Inoculation of suspected material into groups of susceptible and immune pigs.
Virus isolation can be carried out in porcine cell lines using homogenates of
spleen and tonsil.
Immunofluorescence and other serological tests like gel diffusion tests were used
successfully for testing suspected tissues for the presence of specific swine fever
precipitinogen. Pancreas was found to give a more marked reaction than the other tissues.
Fluorescent antibody technique is the most useful test for detecting virus both in field
specimen and infected cell cultures particularly when there are no CPE in the infected
culture.

Control
i. Simultaneous vaccination with virulent swine fever virus and high titre
antiserum is still practiced in some countries.
ii. Attenuated live virus vaccines modified by serial passages in rabbits, swines or
tissue cultures are most frequently used.
iii. Inactivated virus vaccines are safe to use and do not clinical disease. Virus
inactivated by crystal violet is widely used for the past 30 years. Most countries prohibit
the feeding of uncooked garbage to pigs.

Bovine Virus Diarrhea - Mucosal disease

(BVD - MD)
Bovine virus diarrhea and mucosal disease are clinically dissimilar disease
syndromes, and were originally described as separate diseases, but they are now known
to have a common viral etiology. The virus can cause both acute disease, bovine viral
diarrhoea (BVD) and a protracted form of illness, mucosal disease, arising from
persistent infection.
BVD may occur at any age. MD is a persistent infection acquired in utero and
characterized by high mortality and low morbidity. '
1. BVD virus can be either noncytopathogenic or cytopathogenic.
 2. Isolation and demonstration from both BVD and MD were serologically similar
and gave the same experimental disease.
3. The clinical evidence that abortions were a constant finding of field outbreaks
and also the demonstration of "transplacental transfer of the virus to the fetus".
4. Early fetal infection with BVD virus may lead to persistent viremia and a
failure to develop antibodies.
5. Persistently viremic animals could subsequently develop MD. Persistently
viremic animals are infected with only non- cytopathogenic virus, whereas those
developing MD are infected with both non-cytopathogenic and cytopathogenic forms.
6. Experimental production of BVD, result of an acute infection in susceptible
cattle with a clinical syndrome like abortion, teratogenic defects, still births and weaker
calves which lasts for a few days with negligible mortality.

MD is almost invariably fatal with low morbidity. It occurs in cattle, which have a
persistent BVD infection acquired as fetuses characterized by a specific immune
tolerance to the infecting virus strains and consequent lack of antibody to it.
In MD, if the fetus infected at 110-120 days of gestation, i.e., before the age of
immnocompetence, the virus become accepted as self and persist through out the life of
the animal. This explains the lack of antibody to persisting virus and the continued state
of immuno tolerance. Sometimes after birth, when the animal is about 6-18 months of
age, super infection of these persistently viremic animals with the cytopathogenic bio-
type may occur causing MD.
Pathogenesis
The virus is usually acquired by the oronasal route and initial replication occurs in
the oronasal mucosal. In the subsequent viraemia, virus spreads throughout the body
either free in the serum or in association with leukocytes.
Clinical features
The clinical and pathologic manifestation of infection in individual cattle vary
with age and pregnancy status. Three situations are considered: postnatal infection in
nonpregnant cattle, infection in pregnant cattle, infection in pregnant cows and persistent
infection in calves and mucosal disease.

I. Postnatal infection of susceptible non pregnant cattle

a. Acute infection
Infection is most common in animals 8-24 months of age. Although calves may
receive antibodies in colostrum, antibody levels decline by 3-8 months of age and
animals can then become infected. Incubation period 5-7 days, pyrexia (40-41°C),
leucopenia, viremia. In majority it is subclinical infection. There may be diarrhea of
explosive nature with very high morbidity but no mortality. Drop in milk yield in dairy
cows, oculo-nasal discharge and mouth ulcers is referred to as BVD. Animals develop
serum neutralizing antibodies which persist life long.
b. Immuno suppression
Virus induces transient but profound immuno suppression. The virus suppresses the
interferon production, affects lymphocyte function, humoral antibody production and
phagocytosis which paves way for many respiratory and other diseases in calves.
c. Venereal infection
Semen from persistently infected bulls by Al are Natural Service infect cows which
results in early embryonic mortality and repeat breeding.

II. Infection of susceptible pregnant cattle

a. Transplacental infection
The infection to the fetus causes abortion, weak calves, undersized calves and
congenitically deformed calves. The virus replicates in almost all the fetal tissues and the
extent of damage is more in actively dividing cells. Regarding the nervous system it
causes cerebellar hypoplasia, dysplasia, cavitation of the cerebrum and retinal
displacement.
b. Immunological competence of the fetus
Infection in early fetal life becomes persistent viral infection in many tissues and organs.
After birth, the calf remains infected for life. These calves excrete the virus in large
quantity and transmit to all the susceptible healthy animals. Therefore, high probability of
development of MD is possible.

Ill Persistently infected cattle

In susceptible healthy herds in which virus was recently introduced, high
proportion of calves in the next calving season become persistently infected. Mortality
rate exceeds 50% in the preslaughter age i.e., 6-18 months age and the most classical
clinical manifestations are that of MD. MD is characterized by pyrexia, anorexia, profuse
watery diarrhoea, nasal discharge, buccal ulceration and sometimes lameness. Death is
within a few days to 3 weeks.

Lesions
Erosive or ulcerative lesions extending from the mouth through esophagus,
forestomachs, abomasums and intestine. In the intestine, discoloration of the mucosal
folds due to hyperemia and haemorrhage may occur, giving a striped appearance to the
lumenal surface. Necrosis of lymphoid tissues occurs, particularly in the intestine-
Peyer’s patches become hemorrhagic and necrotic.

Diagnosis
History of the herd, abortion, excessive neonatal mortality, congenitally abnormal
calves, episodes of enteric disease and undiagnosed deaths in growing calves.
Viral isolation in cell cultures
Paired sera samples-SNT
FAT, ELISA and DNA Probes.

Control
Strict managemental practices.
Replacement animals especially breeding bulls.
Cattle should not be allowed to mix with sheep and goats.

Vaccination
i. Modified Live BVD vaccine
 ii. Killed BVD vaccine, which is safer for administration even to pregnant cattle.
A booster dose after 3 to 4 weeks of initial vaccination.
Animals should be vaccinated at 5-6 months of age when there is decline in
colostral immunity. For heifers to be done just before breeding and booster one month
before calving or in the last trimester.

Border Disease
(Hairy shaker disease)
Cause
Border disease virus. The virus of border disease is antigenically closely related to the
viruses of bovine viral diarrhea and hog cholera.
Occurrence
Border disease of sheep was first described from the border region of Wales and England.
It occurs in sheep flocks in Great Britain, New Zealand, Australia, and other countries,
including the United States and Canada. Kids (goats) are susceptible and the disease has
been seen infrequently in calves.
Transmission
The virus is shed in excretions and secretions. Spread is by direct or indirect contact.
Clinical & Pathologic Features
The virus causes no overt clinical signs in adult sheep, but infection of pregnant ewes
prior to ~80 days of gestation results in fetal infections with much the same consequences
observed in bovine fetuses infected with BVD virus (see above). Fetal resorption,
abortion, and the birth of persistently infected animals are common sequelae. Persistently
infected lambs often have abnormally hairy birthcoats and congenital tremors resulting
from defective myelination of CNS. Persistently infected lambs that are severely affected
usually die. Those minimally affected may live and provide a source of infection for
other animals.
Diagnosis
 Clinical specimens: Whole blood and serum; affected lambs for necropsy.
 Clinical signs and history are diagnostic.
 The histological finding of nerve fibers with defective myelin sheaths is
supportive.
 The virus can be propagated in a variety of cell cultures but without observable
cytopathic effects. Viral antigen can be demonstrated in infected cell cultures,
blood smears (precolostral blood preferred), and affected tissues (frozen sections)
by immunofluorescence.
 Virus neutralization and ELISA can be used on serum samples to determine the
degree of flock infection.
Prevention
 Prevention is best accomplished by maintaining closed flocks. Efforts to control
the disease involve serologically testing (ELISA, virus neutralization) of all
animals and removing those that are positive. Only negative animals are admitted
to the flock.
 If eradication is not feasible, breeding animals should be exposed to persistently
infected individuals at least two months before breeding.
 An inactivated, adjuvanted vaccine containing BDV and BVDV-1 is used.
 Flavivirus

Louping Ill
(Ovine encephalomyelitis)
Cause
Louping ill virus. Four subtypes are recognized: Spanish, British, Irish and Turkish.
Occurrence
Louping ill is a tick-borne disease, principally of sheep, that occurs in England, Ireland,
and Scotland and in some other European countries. The hosts are sheep and less often,
cattle, deer, horses, pigs, wild rodents and humans. Wild rodents, deer, shrews and grouse
may be naturally infected without clinical signs of disease.
Transmission
The vector and only reservoir is the tick Ixodes ricinus.
Clinical & Pathologic Features
The lymph nodes are initially infected. A viremia follows with spread to other tissues
including in some animals the CNS.
Louping ill is characterized clinically by a biphasic fever and progressive central nervous
system dysfunction. Initial clinical signs are those of high fever and depression of about
24 - 48 hours duration. A second febrile response occurs a few days later accompanied by
signs of central nervous system dysfunction including excitability, incoordination,
muscular tremors, and paralysis. The mortality rate is high in those animals that develop
CNS disease. Some animals recover but display mild neurological signs.
Diagnosis
 Clinical specimens: Fresh and formalin fixed brain tissue.
 A presumptive diagnosis is based on clinical signs and history. Histologic lesions
of meningoencephalomyelitis are supportive.
 The virus can be isolated in young mice inoculated intracerebrally and in cell
cultures.
 The presence of specific antibodies is diagnostically significant. Serological
procedures include ELISA, complement fixation, hemagglutination inhibition
(virus agglutinates goose red cells), gel diffusion and indirect
immunofluorescence.
Prevention
 Inactivated tissue culture vaccines are used in areas where the virus is endemic.
The colostrum of vaccinated ewes protects lambs for a year.
 Animals are dipped, sprayed, etc. in an effort to control ticks.
Public Health Significance
This virus can cause severe encephalomyelitis in humans. Humans may be infected by
the bite of infected ticks, aerosol or contact with infected carcasses. Few cases have been
attributed to ticks and infected animals; most have occurred in laboratory workers.

West Nile Virus Infection

Cause
West Nile virus. It belongs in the Japanese encephalitis serogroup.
Occurrence
West Nile virus infection (WNVI) occurs in countries of Asia, Africa, Europe and North
America. The virus was introduced to the USA in 1999 and by 2004 it had spread
throughout the continental USA. There were >4000 human cases in 2002, with 274
deaths, mainly in people over 60 years of age. Less than 1% of those infected are
symptomatic. The number of cases will probably decrease as the infection becomes
endemic.
As human cases occurred there were many reports of the disease affecting horses in the
USA. More than 15,000 cases were reported in 2002 and somewhat less in 2003. Older
horses are more susceptible.
In 1997 in Israel a neuroparalytic disease of young geese was attributed to WNV. Geese
appear to be the only natural host among domestic avian species.
Transmission
It is estimated that at least 58 mosquito species can carry the virus. Culex pipiens is
considered the most important for maintaining the virus. Upwards of 300 bird species
harbor the virus. House sparrows are thought to be the most important in dissemination.
They are readily infected and have high levels of virus. Although some birds such as
crows may die of infection house sparrows don’t.
Clinical & Pathologic Features
The incubation period in horses is 7 - 14 days. Signs may appear suddenly or gradually;
they may include incoordination, dragging hooves, buckling at knees, difficulty eating
and drinking, stumbling, muscle weakness, dullness, somnolence, paralysis, inability to
rise. Some horses may have a mild infection with low fever, muscle trembling and
evidence of a less severe disease.
Many horses that recover from the disease have residual abnormalities such as
irregularities of gait, behavioral changes and neurological deficits six months after
diagnosis. If horses don’t recover sufficiently they may have to be euthanized. The
mortality rate in unvaccinated horses developing clinical disease is ~33%. Horses
surviving infection are considered immune for life.
The lesions in horses are very similar to those of eastern equine encephalitis (EEE).
There is perivascular lymphocyte cuffs, gliosis and neuronal degeneration. What is
distinctive with West Nile infection is a poliomyelitis affecting the gray matter of the
spinal cord. The brainstem and midbrain are also affected but not usually the cerebral
cortex. In EEE the whole brain is affected with the cerebrum most affected.
Diagnosis
 Clinical specimens: Whole blood collected during the febrile stage and brain
tissue from horses that have died. Acute and convalescent sera.
 A presumptive diagnosis is often based on clinical signs and the microscopic
brain lesions referred to above.
 A definitive diagnosis can be obtained by demonstrating a significant increase in
specific antibody between acute and convalescent sera.
 The virus can be propagated on the chorioallantoic membrane, where it produces
plaques, and in various cell cultures.
 A presumptive diagnosis can be made on the basis of clinical signs and results of
a single serum sample, if those results are positive and the horse has not been
vaccinated.
Treatment
 Steroids to reduce inflammation. General supportive care.
 An equine product containing specific viral antibody is being used in treatment
although its value is questionable.
Prevention
 Vaccination is recommended. A DNA vaccine (first of its kind licensed in the
USA) is available for the prevention of the equine disease. One inactivated
vaccine is given in two doses three to six weeks apart. A recombinant vaccine
using a canarypox vector is also available. Some practitioners vaccinate semi-
annually where mosquitoes are present year-round.
 Strict mosquito control greatly reduces the chances of exposure. Keep horses
indoors from dusk to dawn when insects are most active; keep lights off during
the evening and keep stable areas clear of birds and poultry.

Japanese Encephalitis

Japanese encephalitis virus is the most important mosquito-borne human

pathogen in Japan, China, Korea, Thailand and other countries of southeastern Asia, and
it has recently extended its range westward into India, Nepal, and Sri Lanka and eastward
into the Pacific islands of Saipan and the northern Marinas.
The mosquito Culex tritaeniorynchus, which breeds in freshwater habitats and
irrigated rice fields, and feeds on birds, swine and humans is the most common vector,
swine are the most abundant species of domestinuously provide generations of
susceptible hosts. The mosquito-swine-mosquito transmission cycle serves as an efficient
mode of virus amplification.
In several Asian countries, Japanese encephalitis infection in swine causes
considerable losses because of a high abortion rate in sows and a high neonatal mortality
rate. Adult swine, horses, cattle, and sheep usually do not manifest clinical disease, but
they may serve as virus amplifiers.
The disease in horses is clinically similar to EEE, WEE, VEE, and Borna disease,
but the mortality rate is relatively low (0 - 10%). Domestic pigs may also be affected.
The virus can be isolated employing cell cultures of ovine origin and the yolk sac of
chicken embryos.
In Japan, control of the human and swine diseases has been very successful; a
national program is based on immunization of children and swine with different
inactivated virus vaccines produced in mice. Attenuated live-virus vaccines for human
and animal use are under development; these vaccines offer the possibility of lower costs
and may be suitable for use in large areas of southeastern Asia.

Wesselsbron Disease
This disease is caused by Wesselbron virus (Flavovirus) a member of the yellow
fever group and affects sheep, cattle and humans. It occurs widely in Africa and has been
reported from Thailand. It is transmitted by mosquitoes. The disease in sheep is
characterized by high fever and death in newborn lambs and abortion in pregnant ewes.
Infection of pregnant cattle may result in abortions and congenital anomalies of the
central nervous system. Flu-like symptoms are noted in human beings. The virus can be
isolated from liver and spleen employing cell cultures of ovine origin and the yolk sac of
chicken embryos. Modified live virus vaccines are used in areas where the virus is
endemic. Mosquito control reduces exposure.

Prions and Transmissible Spongiform Encephalopathies

Prions are proteinaceous infectious particles that cause chronic degenerative,

neurological, invariably fatal diseases of animals and humans referred to as transmissible
spongiform encephalopathies (TSE). The animal TSEs are the following:
Scrapie
Bovine spongiform encephalopathy
Feline spongiform encephalopathy
Transmissible mink encephalopathy
Chronic wasting disease of mule deer and elk
Spongiform encephalopathy in captive ruminants
Historical
Scrapie has been recognized as a distinct disease entity for at least 300 years (it was
formally recognized in England in 1732). The nature and etiology of Kuru, a human TSE
transmitted by ritual cannibalism, was elucidated in the 1970s. The human TSE
Creutzfeldt-Jakob disease (CJD) has been known for many years. Bovine spongiform
encephalopathy (BSE) was recognized in Britain in 1986.
William Hadlow is credited as the first to make the connection between kuru and scrapie.
Carleton Gajdusek received the Nobel Prize in 1976 for his work correlating human (kuru
and Creutzfeldt-Jakob disease) and animal TSEs. Prusiner was awarded the Nobel prize
in 1997 for his discovery of prions. He had introduced the appellation "prion" in 1982.*

Prion Characteristics
The proteins involved are thought to be derived from a post-translational modification
of a normal cellular protein with unknown function. The normal cellular protein is
designated PrP and the prion PrP with a superscript associated with a particular
disease, for example PrPsc in scrapie.
The protein involved is approximately 254 amino acids in length. Its function is not
known. In humans, the PrP gene is located on the short arm of chromosome 20.
They multiply or reproduce by converting the normal cellular protein PrP to the PrP sc
the infectious form via a conformational change.
Prions are particularly resistant to: 135°C for 18 minutes, ionizing radiation,
ultraviolet light, concentrations of formaldehyde that kill viruses, and nonionic
and non-denaturing anionic detergents. They are sensitive to urea, phenol, sodium
dodecyl sulfate (SDS), sodium hypochlorite and other agents that denature
proteins.
Prions do not elicit antibodies in the host they infect; however, antibodies can be
raised against them in other animals.
Agents of TSEs can be transmitted to laboratory animals by various injection routes.
However, prions show a marked restriction of host range. For example, PrP sc will
readily passage in sheep, but will only rarely infect other species such as mice,
hamsters, and certain primates. All attempts to transmit scrapie to guinea pigs,
rats, and chimpanzees have failed to date. Maintenance of prions in cell culture is
 also very difficult, although low titers of PrP sc have been maintained in a mouse L
cell line.
Prions are partially resistant to proteases and have strong tendency to form aggregates
or polymers.
Antibodies to prions allowed the visualization of rod structures in infected brains,
which consisted of filaments or fibrils made up of prion protein. Amyloid plaques
were shown to be made up of these fibrils.
Amyloid plaques and fibrils are located between nerve cells, presumably released
following destruction of cells.

Transmissible Spongiform Encephalopathies of Animals

Scrapie
Cause
A prion. Several polymorphisms of the PrP gene have been identified and associated with
differences in susceptibility to scrapie in breeds of sheep. These polymorphisms result in
the production of amino acid substitutions in PrP.
Occurrence
This fatal neurological disease affects sheep and goats. The disease is endemic in the
United Kingdom, but less common in other European countries. It has been eradicated
from Australia and New Zealand. It occurs rarely in North and South America. The
disease was first diagnosed in the United States in Michigan in 1947.
Transmission
The agent of scrapie is transmitted both vertically in family lines of flocks and
horizontally by direct or indirect contact with infected placenta (e.g., ingestion,
abrasions).
Pathogenesis
The scrapie prion is found in the spleen, mesenteric and retropharyngeal lymph nodes,
placenta, intestine, ovary and palatine tonsil where it replicates and reaches the CNS via
fibers of the autonomic nervous system. As the disease progresses high concentrations of
the infectious prion accumulate in the brain. When clinical signs appear death usually
occurs within six months.
Clinical & Pathologic Features
Scrapie is a non-febrile, insidious disease with a long incubation period with the peak
incidence of signs at 3 - 4 years. In infected animals the agent may be detected in
lymphoreticular tissues as early as eight months of age and in the CNS at about two
years. Clinical signs may appear as early as three and a half years. Initial signs are
restlessness, excitability, and grinding of the teeth. Later signs are tremors, intense
pruritus resulting in shedding of wool and laceration of the skin, and convulsions. Death
occurs from several weeks to several months after onset of signs.
Changes are seen microscopically in neurons of the medulla, pons, midbrain and spinal
cord. There are single or multiple vacuoles surrounded by zones of cytoplasmic
degeneration in neurons. Similar spongiform changes may also be seen in the neuropil of
the CNS and peripheral nervous system. Fibrils and amyloid plaques are commonly seen
in the brain.
Diagnosis
Clinical specimens: Brain and spinal cord.
Clinical signs are suggestive of scrapie but definitive diagnosis requires histological
examination of brain tissue.
Supportive is finding characteristic fibrils with electron microscopy.
Other procedures used to confirm the diagnosis are immunoblotting to detect PrP sc,
and immunohistochemical staining of PrPsc.
Some of the methods used for the diagnosis of variant CJD, including demonstration
of the agent in tissue of the palatine tonsil, are being explored.
Prevention
 This is best accomplished by removing affected animals and maintaining closed
flocks.
 In countries where the disease is notifiable, affected flocks are quarantined and
slaughtered (particularly bloodline-related animals) or depopulation is carried out.
 Measures are implemented to prevent importation of infected animals.
 In some countries, the frequency of the disease is such that only gradual
elimination is feasible.
 Genetics is being used as a tool in eradication of scrapie. This is accomplished by
eliminating susceptible bloodlines. Several polymorphisms of the PrP at several
codons are associated with resistance to the disease. No such polymorphisms have
been identified in goats.

Bovine Spongiform Encephalopathy

(Mad cow disease)
Cause
A prion that is not considered host species-specific. It has been concluded that the prion
involved was acquired by eating contaminated (scrapie prion) meat and bone meal
derived from slaughterhouse offal. A change had been made in the rendering process that
allowed survival of the scrapie agent. Factors involved in the extent of the epidemic were
no doubt the fact of the endemicity of scrapie and large proportion of sheep tissues and
offal in the meat-bone meal supplement. Subsequently the supplement also contained
offal and tissues from cattle with BSE.
Occurrence
It is estimated that more than 170,000 cases were confirmed in Britain prior to
eradication. More than 99% of the laboratory confirmed cases of BSE have occurred in
Great Britain. A small number of cases attributed to the importation of British cattle were
identified in Switzerland, France, Portugal and Ireland. Two cases have been identified in
Canada and two cases in the USA, one of which was traced to an affected herd in
Canada.
Transmission
This occurred orally through the eating of the meat-bone meal supplement referred to
above. There was no evidence of horizontal or vertical transmission.
Clinical & Pathologic Features
The pathogenesis is probably roughly analogous to other TSEs, but details are not yet
clear. Following exposure to the agent of BSE, there is a long incubation period prior to
development of clinical signs. The average time appears to be about four to five years.
Affected cattle display clinical signs associated with progressive CNS dysfunction,
including behavioral changes, incoordination and hypersensitivity to various stimuli. The
disease is uniformly fatal within 1 - 6 months after the onset of signs.
Diagnosis
 Clinical specimens: Fresh or formalin-fixed brain tissue.
 Diagnosis of BSE is based on clinical signs and the subsequent demonstration of
the characteristic spongiform changes in the brain by histopathologic
examination.
 Prion fibrils can be observed in brain extracts with electron microscopy.
 Immunohistochemical staining of tissues for the prion protein.
 Immunoblotting employing monoclonal antibodies to demonstrate the proteinase
K resistant prion of BSE.
 There is an urgent need for a reliable live animal diagnostic test. One approach
has involved olfactory and tonsillar biopsies and monoclonal antibodies.
 The rapid screening test used by the United States Department of Agriculture is
the TeSeE® test (Bio-Rad Laboratories), which is an ELISA-based test for the
BSE-specific prion protein. The company also produces similar tests for the
detection of chronic wasting disease and scrapie.
Prevention
 All protein derived from ruminants must be excluded from cattle feed.
 BSE is a reportable disease in most countries. Its occurrence, even one case, can
have a devastating effect on the cattle industry. Quarantine and slaughter of the
herd is usually carried out.
 Affected cattle are incinerated. Decontamination of premises is carried out with a
strong solution of sodium hydroxide or sodium hypochlorite.

Feline Spongiform Encephalopathy

Cause
This is presumed to be the same as that causing BSE and scrapie of sheep and goats, viz.
a self-replicating protein, referred to as a prion.
Occurrence
Since 1990 more than 80 cases of feline spongiform encephalopathy (FSE) have been
reported from Great Britain. One case has been reported from each of the following
countries: Northern Ireland, Norway, and Liechtenstein. Nine cases have involved
cheetahs and two in lions. Three of the nine cases in cheetahs occurred abroad but
originated in Britain. FSE has not been reported from North, South, or Central America.
Given the strict regulations for the control of scrapie and BSE the disease in cats, even in
Great Britain, should markedly decline.
Transmission
It is presumed that cats acquire the infectious agent by eating infectious bovine, ovine, or
goat tissue most likely in prepared cat foods, raw meat or table scraps. It is not known if
transplacental transmission takes place and it seems unlikely that cat-to-cat transmission
occurs.
Clinical Features
The incubation period is not known, but is presumed to be long as is the clinical course.
The disease is seen most frequently in cats 2 - 7 years of age. Signs include behavioral
changes, salivation, hyperesthesia, muscular tremors and ataxia. All cases terminate
fatally.
Diagnosis
 Clinical specimens: Brain and spinal cord.
 Although the long course and clinical signs may suggest FSE, a definitive
diagnosis can only be made after necropsy.
 The changes observed in the brains of cats with FSE are indistinguishable from
those seen in brains of cattle with BSE. The former differs somewhat from the
changes seen in sheep with scrapie. Characteristic histological changes include
vacuolation of the grey matter neuropil and neurons. Fibrils of the kind seen in the
brain in scrapie and BSE can be seen in the brain of cats with FSE by electron
microscopy.
 Detection of the prion protein involved is not carried out in most veterinary
diagnostic laboratories.
Prevention
With the near eradication of BSE it seems very unlikely that its prion will be found in cat
food.

Transmissible Encephalopathy of Mink

Transmissible encephalopathy of mink (TEM) is an insidious neurologic disease
characterized by a long incubation period (eight months or longer) and clinical signs of
hyperirritability, loss of weight, ataxia, compulsive biting, somnolence, and ultimately
death in about 2 - 6 weeks.
There are no gross necropsy lesions, but characteristic spongiform changes in the brain
are noted histologically. These spongiform changes are essentially identical to those
observed in the brain of sheep with scrapie and with the spongiform encephalopathies of
other animals, including cattle. Mink are thought to be infected by eating contaminated
meat from ruminants. Cannibalism and fighting may spread the agent among other mink.

Chronic Wasting Disease of Deer and Elk

Chronic wasting disease (CWD) was first recognized as a clinical syndrome in a farm-
reared mule deer in Colorado in 1967. That the disease was a TSE was confirmed in
1978. Clinical features and pathology were similar to other animal TSEs. The disease has
subsequently been seen in captive and wild deer, and elk in western states of the U.S.,
Wisconsin and Minnesota, and two western Canadian provinces. Like other TSEs the
incubation period is long with progressive loss of condition, striking weight loss and
always followed by death.
Efforts are being made to eliminate the disease from some herds by quarantine and
removal of positive animals.
As yet, there in no evidence of transmission of the disease to humans or to domestic
cattle and sheep.

Spongiform Encephalopathy in Captive Ruminants

During the outbreak of BSE in Britain in 1986, spongiform encephalopathy was observed
in some captive wild ruminants including nyala, oryx and greater kudu. These cases were
attributed to the feeding of the same kind of dietary supplement that gave rise to BSE*.
 Human Transmissible Spongiform Encephalopathies
 Creutzfeld-Jacob Disease (CJD)
 Variant CJD (vCJD)
 Kuru
 Gertsmann-Straussler-Sheinker Syndrome
 Fatal Familial Insomnia

Creutzfeld-Jacob Disease
This human TSE is rare with a worldwide incidence of approximately one in a million. It
appears spontaneously, but also with a familial association of ~10%. The means of
transmission, except for iatrogenic transmission, is unknown. Means of iatrogenic
transmission include corneal transplant, via intracerebral electrodes, surgical operations
with contaminated instruments, dura mater grafts, and treatment of children and teenagers
with growth hormone extracted from the hypophysis (pituitary gland) of people dying of
CJD. Most cases are seen in people 50 - 70 years of age. The course of the disease is
about six months and the average age of death is 58.

Variant Creutzfeld-Jacob Disease (vCJD)

This is the human TSE attributed to consumption of meat from cattle with BSE. It was
identified in Britain in 1996. However, the mode of transmission and the factors that
affect vCJD susceptibility are not understood. Ways in which vCJD differs from CJD are:
 Early age of onset, an average of 27 years as compared to > 50 for CJD.
 Presentation is psychiatric as opposed to neurological symptoms.
 The typical electroencephalogram appearances of CJD are absent.
 In vCJD flower-like amyloid plaques are seen.
 The period of illness prior to death is 13 months compared to three months for
CJD.
 In the UK from 1994 through 2003, 104 deaths were confirmed to be from vCJD.
However, the number of reported cases has been decreasing in recent years. In the
USA, CJD deaths among persons < 30 years of age are fewer than five deaths per
billion per year.
Procedures for the diagnosis of vCJD are the same as those used for the diagnosis of
BSE.
Kuru
This human TSE occurred among the Fore tribes of New Guinea as a result of ingesting
human brains of the dead during ritual cannibalism, a rite of mourning for their dead.
Death occurred within a year after the onset of symptoms.

Gertsmann-Straussler-Sheinker Syndrome
This is a very rare form of CJD that appears to result from a mutation in the prion gene.

Fatal Familial Insomnia

This human TSE is considered a variant of CJD. It was found recently in patients with a
prion gene mutation of an amino acid change at position 178. The first symptoms are loss
of sleep, followed by hallucinations and death in less than a year.
 Hepadnaviridae

This is a family of small double-stranded DNA viruses with an icosahedral capsid

covered by an envelope. They infect herons, ducks, woodchucks, squirrels and humans
(the important hepatitis B virus), but are of little veterinary significance.
Viral Characteristics
 These double-shelled viruses have an icosahedral capsid covered by an envelope
(40 - 48 nm in diameter).
 They have a circular DNA genome, which is partially double-stranded (~ three-
fourths) and partially single-stranded (~ one-fourth).
 They require a viral encoded reverse transcriptase to replicate in that the virion
DNA is synthesized from an intermediate RNA template.
 The DNA may be integrated into the host DNA of the cell during replication.
 These viruses are highly host specific, have a predilection for the liver and
produce persistent infections.
Viruses of the Hepadnaviridae are placed in genera based on differences in genomic
structure and on whether they are mammalian viruses (Orthohepadnavirus) or avian
viruses (Avihepadnavirus).
Significance
Orthohepadnaviruses infect woodchucks and ground squirrels and one species, hepatitis
B virus, causes the important human disease, hepatitis B (HB), which is often referred to
as serum hepatitis.
Animal hepadnaviruses include viruses of captive woodchucks, ground squirrels and
ducks. In recent years, other hepadnaviruses have been recovered from wild and domestic
species such as herons, geese, marsupials and orangutans. The avian viruses can be
propagated in fetal duck liver hepatocytes.
One species, duck hepatitis B virus, causes a worldwide natural infection of Anas species
but not of Muscovy ducks. The infection is acquired congenitally and produces a chronic
viremia. It does not appear to be economically significant and shouldn’t be confused with
the duck hepatitis caused by types of Picornavirus.
It is of interest that duck hepatitis B virus is being used as an experimental model for
studies of hepadnavirus infection and replication.
HBV is among the most important human pathogens and is thought to infect chronically
over 300 million people worldwide. Acute or chronic hepatitis, asymptomatic persistent
infection, cirrhosis and hepatocellular carcinomas are among the consequences of HBV
infection.

Deltavirus
Hepatitis delta virus (the only species) infection; complicates hepatitis B infection. It is of
interest that hepatitis delta virus is distinct from all other viruses in resembling in some
features viroids and satellite viruses found in plants.