MSC Thesis 10
MSC Thesis 10
MSC Thesis 10
By
JULY 2005
DEBRE ZEIT, ETHIOPIA
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TABLE OF CONTENTS
Page
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2.3.3.2 Age of the host .................................................................................................. 14
2.3.3.3 Genetic factor of the host .................................................................................. 15
2.3.3.4 Arrested development of the parasite in the host (Hypobiosis) ........................ 16
2.3.3.5 Difference in the strain of parasite .................................................................... 16
2.4 DIAGNOSIS ....................................................................................................................... 17
2.4.1 Laboratory diagnosis................................................................................................ 17
2.4.1.1 Faecal examination ........................................................................................... 17
2.4.1.1.1 Qualitative methods ................................................................................... 18
2.4.1.1.1.1 Direct faecal smear examination......................................................... 18
2.4.1.1.2 Concentration techniques ........................................................................... 18
2.4.1.1.3 Quantitative methods/Egg counting techniques ........................................ 18
2.4.1.2 Faecal culturing................................................................................................. 19
2.4.1.3 Sentinel worm counts........................................................................................ 19
3. MATERIALS AND METHODS ....................................................................................... 20
3.1 STUDY AREA .................................................................................................................... 20
3.2 STUDY POPULATION ........................................................................................................ 24
3.2.1 Study animals ........................................................................................................... 24
3.2.2 Sample size .............................................................................................................. 25
3.3 STUDY METHODOLOGY .................................................................................................... 26
3.3.1 Study type ................................................................................................................ 26
3.3.2 Study procedures...................................................................................................... 26
3.1.1 Statistical analysis methods ..................................................................................... 28
4. RESULTS OF THE STUDY .............................................................................................. 29
4.1 POST MORTEM RESULTS ................................................................................................... 29
4.1.1 Nematode species identification and their prevalence ............................................. 29
4.1.2 Post mortem findings based on season, agro-ecology, age, species & sex
differences. ........................................................................................................................ 29
4.1.3 Correlation of total nematode count to total epg counts from slaughtered
sheep and goats. ................................................................................................................ 37
4.2 COPROLOGICAL RESULTS ................................................................................................. 38
4.2.1 Prevalence of nematode infection from faecal examination.................................... 38
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4.2.2 Results of faecal egg count ...................................................................................... 40
4.2.2.1 Fecal egg counts based on season, agro-ecology, age, species and sex
differences. .................................................................................................................... 40
5. DISCUSSION ...................................................................................................................... 43
5.1 IDENTIFICATION OF NEMATODE SPECIES........................................................................... 43
5.2 NEMATODE PREVALENCE ................................................................................................. 43
5.3 SEASONAL DYNAMICS IN PREVALENCE ............................................................................ 45
5.4 INTENSITY AND TYPE OF INFECTION ................................................................................. 46
5.5 CORRELATION OF EPG TO NEMATODE BURDEN ................................................................. 48
6. CONCLUSION AND RECOMMENDATIONS .............................................................. 49
7. REFERENCES.................................................................................................................... 51
8. ANNEXES ........................................................................................................................... 66
9. CURRICULUM VITAE.................................................................................................... 84
10. SIGNED DECLARATION SHEET ................................................................................ 86
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LIST OF TABLES Page
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LIST OF FIGURES Page
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LIST OF ANNEXES Page
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LIST OF ABBREVIATIONS
Buno: Bunostomum
Chab :Chabertia
Oesophago: Oesophagostomum
Spp.: Species
Tricho: Trichostrongylus
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ACKNOWLEDGEMENT
First of all tanks be to my heavenly Father for His guidance of my life according to His will.
I would like to express my heart-felt gratitude to my advisor Dr Abebe Wossene for his high
standard guidance, valuable and all round assistance in this research.
Southern Nation, Nationalities and Peoples Regional State for allowing me to follow
this postgraduate program.
Dr Zerihun W/ Senbet, Dr Nejib Mohammed, Ato Debebe Deboch and Wro. Asrat, the
staff members of Bonke and Mirab-Abaya wereda Bureau of agriculture for the help
during my field work at the weredas.
Dr Aseged Bogale, Dr Fikre Lobago and Dr Boja Hindabu for their technical
assistance and encouragement during paper writing and analysis of the results.
I would like offering my special thanks for the lovely sacrifice paid by my family during my
educational life.
The prayer and patience of my wife, Wro. Bayoush Getachew, my daughters; Deborah,
Eminet and Bersabeh during my absence for such a long period is also appreciated.
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ABSTRACT
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Out of 98.9% of infected animals in the study area, 97.7% were infected by more than one
nematode species. 64.0% of poly-parasitised animals in the study area were infected by 4 or
more nematode species. 58.4% of the infected animals harboured light infestation, 40.5% were
moderate and only 1.1% was severely infected by gastrointestinal tract nematode species. The
mean total nematode burden and mean total epg count both from slaughtered animals in this
study was positively correlated (Spearman’s rho (rs)=0.57, p<0.001) and the correlation of
mean epg to individual nematode species burden was significant for three nematode species;
Trichostrongylus colubriformis, (rs=0.50); Haemonchus contortus, (rs=0.45) and
Oesophagostomum columbianum, (rs=0.39) with p<0.001 for all correlations. Faecal
examination results from population of sheep and goats during four sampling periods indicated
significant seasonal variation (p<0.05) for all study sites. Majority of infected animals had a
faecal egg count in the range of 50-800 epg and only few proportions of animals had faecal
egg count over 1200.
The widespread existence of various nematodes in the form of polyparasitism both in sheep
and goats in all agro-ecology of the study area, and the high nematode burden both at
postmortem and coproscopic examination suggest the institution of various control measures
including strategic anthelmintic treatment for efficient utilization of the available small
ruminant resources at hand.
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1. INTRODUCTION AND OBJECTIVES
1.1 Introduction
The contribution of livestock to the human being particularly in the developing countries is
numerous and small ruminant production is an important component of livestock production in
Africa (Rege, 1994). Small ruminants in Africa represent 21% of the world’s small ruminant
population (sheep, 17% and goats, 30% of the world’s sheep and goats population
respectively) (Ibrahim, 1998).
Highly adaptive nature to range of environments, ability to utilize wide variety of plant
species, short generation cycle and high reproductive rates which lead to high production
efficiency made sheep and goats complementary to cattle and camel (Rege, 1994; Ibrahim,
1998). Sheep and goat are a major source of income (cash) and food protein for rural farmers
in most part of tropics including Ethiopia (Ibrahim, 1998), and skin, fiber, manure and as an
investment (Devendra and Mcleory, 1982; Rege, 1994). However, the full exploitation of
these resources is hindered in the tropical environment, and particularly in Africa due to a
combination of factors such as drought, poor genetic potential of the animals, traditional
system of husbandry and the presence of numerous prevalent diseases (Schillhorn van veen,
1985; Mboera and Kitalyi, 1994; Mtenga et al., 1994; Ndamukong, 1994; Rege, 1994;
Ibrahim, 1998).
Among the diseases that affect small ruminants; gastrointestinal tract (GI) nematode parasites
impose severe economic impact on sheep and goat production in the world exerting
deleterious effect on host, which may be manifested by lowered vitality, reduced rate of
reproduction, slower growth rate or death of infected individuals (Wilford, 1974; Belschner,
1976; Smith and Sherman, 1994; Kaufmann, 1996; Ibrahim, 1998) and it is a serious problem
of the developing nations of the world, particularly, where nutrition and sanitation are poor
(Mostofa et al., 1996).
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The severity of the disease and effects it cause is prominent in young flocks (lambs) and
yearlings (Belschner, 1976) and to those individuals that are stressed (Soulsby, 1982; Urquhart
et al., 1996; Radostits et al., 2000). Hence the disease deserves special attention as it
diminishes the capacity of sheep and goats to achieve their inherent potential level of
production for any given feeding and management regimen (Morris, 1988).
Various epidemiological factors are affecting the distribution and importance of parasitic
disease. One of the factors seriously affecting the pre-parasitic phase of nematode
development is the presence of suitable climate, particularly of temperature and moisture. The
requirements of the different free-living stages of nematodes in this regard vary from parasites
to parasites. As a result some parasites are more adapted to temperate cool environment while
others are adapted to warm tropical environment (Craig, 1998).
Some of the major nematodes responsible for GIT parasitosis in small ruminants under
tropical environment are: Haemonchus, Trichostrongylus, Nematodirus and Cooperia spp.
(Family: Trichostrongylidae); Bunostomum trigonocephalum and Gaigeria pachyscelis
(Ancylostomatidae); Chabertia ovina and Oesophagostomum spp (Strongylidae); and
Trichuris and Strongyloides spp. (Troncy, 1989). A number of published works indicate that
these nematode species are infecting sheep and goat population of tropical environment, either
singly or mixed in a host at a time. However, under natural field condition, polyparasitism
predominates in most cases and the prevalence varies depending up on the area and the
parasite species involved.
In Ethiopia, a number of attempts have been made so far to identify the different genera and
species of nematodes in small ruminants and determine the prevalence both by coprology and
post mortem examination. The report made by Graber (1975) was one of the earliest and
extensive attempts in describing the different types of helminth parasites of domestic animals
including that of small ruminants from different parts of Ethiopia. The existence of H.
contortus, Oe. columbianum, S. papillosus, Oe. venulosum, C. ovina, Skrjabinema ovis, B.
trigonocephalum, Trichostrongylus spp., Teladorsagia spp. and Trichuris spp. was reported
2
from Oromiya (Yabello, Debre Zeit, Bale); SNNPRS (Sidamo, Gamo Gofa); Amhara (Debre
Berhan, Wollo, Kombolcha, Shoa) and Somale region (Ogaden) (Graber, 1973; Graber, 1975).
A valuable input in identifying parasites of small ruminants was also made through externship
program by students of the Faculty of Veterinary Medicine of Addis Ababa University and by
some research institutes and regional laboratories of the Ministry of Agriculture. In most
cases, five genera of nematode parasites from different angles of the country were reported,
and these include: Haemonchus spp., Oesophagostomum spp., Bunostomum spp.,
Trichostrongylus spp. and Trichuris spp.. Longistrongylus elongata was reported in sheep at
Debre Berhan, ILRI research station (Tembely et al., 1997). Skrjabinema ovis was reported in
sheep at Debre Berhan(Tembely et al., 1997) and from sheep and goats of eastern part of
Ethiopia (Abebe and Esayas, 2001). Chabertia ovina was recovered from sheep bought from
Arsi and Wollo areas and slaughtered at Addis Ababa abattoir (Bekele et al., 1982). Details of
findings and the specific areas so far studied are presented in Annex (4 & 5).
Although extensive studies are lacking in southern part of Ethiopia, two preliminary studies by
the undergraduate students of the FVM of AAU at Wolayta and its surrounding indicated the
importance of gastrointestinal tract nematodes as a major problem to sheep and goat
production of the study sites (80 to 90% prevalence according to Dereje, 1992 and Haileleul,
2002). However, detail studies covering all agro-ecological zones and the important
epidemiological factors that may have great role in the disease control are lacking. Therefore,
the general purpose of this study is to identify GIT nematode species existing and to observe
the major epidemiological factors that determine occurrence and magnitude of GIT
nematodiosis of small ruminants in three different agro-ecological zones of Southern Ethiopia.
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1.2 Objectives of the study
A. To identify the nematode species parasitising sheep and goats and determine their
prevalence in three different agro-ecological zones of the study areas.
B. To observe the seasonal dynamics of GIT nematodiosis of small ruminants in the study
areas and identify the involved epidemiological factors associated with the GIT nematodiosis.
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2. LITERATURE REVIEW
The effect of parasitism on individual animal (host) is not distinctly manifested clinically for
specific parasite species. In general, all intestinal nematodiosis lead to reduced feed intake
which may be partly due to increased production of cholecystokinin by the intestinal cells,
absorbed into blood system and act on neuro-endocrine control centre and suppress the
appetite. However, based on the predominating clinical symptoms, we can group
gastrointestinal tract nematodes as those that cause decreased feed utilization by the host,
those that cause the destruction of host’s tissues and those that cause loss of blood (Wilford,
1974; Soulsby, 1982; Troncy, 1989; Fox, 1997).
2.1.1 GIT nematodes that cause decreased feed utilization by the host
This group includes mainly Teladorsagia spp., Trichostrongylus spp., Cooperia spp.,
Strongyloides papillosis, Oesophagostomum spp and Nematodirus spp.. The pathological
changes observed due to these parasites include: tunnel under mucosal epithelium of
abomasum (Teladorsagia spp., Trichostrongylus axei) and small intestine, erosion of mucosal
epithelium of abomasum and intestine, catarrhal enteritis, villous atrophy in anterior small
intestine, hyperaemia and oedema. In extreme cases, diphtheritic enteritis and exudates hinder
absorption (Dunn, 1978; Soulsby, 1982; Urquhart et al., 1996; Radostits et al., 2000).
5
The manifestation of these lesions is marked alteration in the influx and efflux of water and
electrolytes (chlorine and sodium ions) in the bowel and morphological and biochemical
changes in the epithelial cells and their microvilli (Soulsby, 1982) leading to protein losing
enteropathy, accompanied by excessive mucus production, diarrhoea, weight loss, hypo
proteinaemia (Smith and Sherman, 1994; Urquhart et al., 1996), pica (Troncy, 1989) and in
chronic cases this form is almost indistinguishable from malnutrition (Dunn, 1978).
2.1.2 GIT nematodes that remove the host’s tissue and blood
This group of parasite includes Haemonchus spp., Chabertia ovina, Gaigeria pachyscelis,
Bunostomum trigonocephalum and Trichuris spp. In most cases, the predominating syndrome
is said to be progressive debilitating anaemia (Smith and Sherman, 1994), but hyper acute and
acute infection of Haemonchus contortus and acute infection of Gaigeria pachyscelis could
lead to death without any more clinical manifestation than acute anemia (Belschner, 1976;
Dunn, 1978; Soulsby, 1982; Troncy 1989; Smith and Sherman, 1994; Bowman, 1999;
Radostits et al., 2000).
The losses caused by Haemonchus contortus are more severe and important due to its extreme
pathogenecity, wide geographical distribution in tropics and its high prevalence in small
ruminants (Dunn, 1978; Troncy, 1989). Gaigeria pachyscelis is a highly virulent nematode
that sucks blood and can cause death of a host even in small burden as few as 24 parasites
(Soulsby, 1982). Chabertia ovina is a plug feeding parasite of large intestine which causes
loss of blood when it draws plugs of mucosa by its wide mouth, and its blood sucking is said
to be accidental, and loss due to haemorrhage at the biting site is voluminous (Soulsby, 1982;
Troncy, 1989). Bunostomum trigonocephalum is a bloodsucker causing progressive anemia
(Dunn, 1978; Soulsby, 1982) and blood-sucking habit of Trichuris ovis was stated by Troncy
(1989).
The clinical manifestations due to these nematodes vary from sub-clinical to bleeding to death
(Dunn, 1978; Soulsby, 1982; Troncy, 1989; Urquhart et al., 1996) and could be generally
6
summarized as pale mucus membrane, oedema on the ventral aspect of the body, bottle jaw,
weakness, wool falls out in patches, prostration and death.
Marked tissue reaction manifested by formation of nodules in the intestine of infected small
ruminants is commonly observed in infestation by the 3rd stage larvae of Oe. columbianum
(Soulsby, 1982; Troncy, 1989; Urquhart et al., 1996). In previously exposed sheep and goats
due to sensitisation, 3rd stage larvae of Oe. columbianum pass into sub mucosa of small
intestine and some times under heavy infection to sub mucosa of large intestine and marked
inflammatory reaction around each larva takes place. These nodules (inflammatory reaction)
could be as big as 2.0 cm in diameter and containing greenish eosinophilic pus and fourth
stage larvae (Urquhart et al., 1996).
According to Soulsby (1982), these nodules are due to leucocytes, especially eosinophils and
foreign body giant cells collected around the parasites and the focus becomes encapsulated by
fibroblasts. Whenever there is massive infestation, the number of nodules will be numerous
while the colon contains few adult worms. Extensive nodules on the intestinal walls and
mucosa interfere with absorption, bowel movement, digestion and when these supurative
nodules rapture to peritoneal surface causing supurative peritonitis and multiple adhesions
(Soulsby, 1982; Smith and Sherman, 1994).
Most GIT nematodes have the same basic life cycle. Majority are oviparous and the eggs are
similar and very characteristic type, and immediate transfer of infection from one host to
another does not occur (Troncy, 1989; Urquhart et al., 1996; Craig, 1998; Hendrix, 1998).
After the eggs passed in faeces of the host, the first stage larva develops with in the egg and
then hatches in the external environment. The first stage larvae feed on bacteria in the faecal
pellet and moults to second stage larvae, during which the larvae shed complete cuticle,
including the lining of mouth opening and the excretory pore. Once the moult is over, the
larva is very active, it starts feeding vigorously on the bacteria with in faecal pellet and enters
7
to the second moulting period. The stoma closes and the larva is sealed off with in the
separate cuticle, and this is ensheathed third stage larva (L3), and it takes 6 to 14 days to reach
to this stage in most cases (Dunn, 1978). This ensheathed L3 can not feed and lives on its
stored food. During this stage, the larvae leave faecal pellet and available on nearby pasture
waiting for a host (Dunn, 1978; Soulsby, 1982; Troncy, 1989; Urquhart et al., 1996; Craig,
1998; Hendrix, 1998; Bowman, 1999).
Though most GIT nematodes follow the above basic life cycle, some nematodes are showing
exceptional circumstances. Nematodirus and Trichuris species produce egg with thick shelled.
In Nematodirus species, the L3 development takes place with in the egg shell, and the infection
of the host is acquired by the ingestion of the L3 with in the egg shell or hatched as L3 and
available on the pasture (Kaufmann, 1996; Urquhart et al., 1996). While in Trichuris species
the infective larvae develop after being passed in faeces with in the egg shell and the animal
will be infected by ingesting the infective larvae inside the egg shell (Kaufmann, 1996). In
Strongyloides species, the eggs when passed in the faeces contain fully developed larvae and
have a possibility of developing either to free living adult or infective third stage larvae. In
some parasites (Bunostomum and Strongyloides) these infective stages in the external
environment may infect the host through skin penetration, i.e. they don’t passively wait for
ingestion (Troncy, 1989; Hendrix, 1998).
In vast majority of nematodes the parasitic phase starts after ingestion of third stage larva (L3).
The retained sheath of infective stage larvae is exsheathed through host and parasite stimuli
(Soulsby, 1982). Then the exsheathed larva come in contact with the mucosa, penetrate deeply
between the villi or into the glands. Most of the subsequent development occurs in, rather
than on, the mucosa. In most cases these larvae emerge to the lumen at fifth stage (L5) though
some superficially developing genera emerge at the late fourth stage (Dunn, 1978; Troncy,
1989; Craig, 1998). In both cases these young adults feed, mature, copulate and become egg-
laying adults. Then, eggs pass to external environment with faeces (Dunn, 1978). The
prepatent period varies between 20 to 40 days in most cases (Troncy, 1989; Kaufmann, 1996;
Urquhart et al., 1996).
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2.3 Factors affecting the epidemiology of GIT Nematodes
Many factors are known to influence the transmission and prevalence of nematode infections
in grazing ruminants (Urquhart et al., 1996) and the disease pattern is determined by factors
influencing the susceptibility of the host, the number of infective larvae accumulating on the
pasture and the number of larvae undergoing hypobiosis (Radostits et al., 2000). Broadly the
three influencing factors that can determine the occurrence of small ruminant gastrointestinal
tract nematodiosis could be mentioned (Smith and Sherman, 1994; Stromberg, 1997).
The environment-host interaction that can influence the occurrence and prevalence of
gastrointestinal tract nematodiosis of small ruminants include:
Nutrition plays great role in worm burden control. Poorly fed animals are more susceptible
and carry more worm burden due to their failure to throw over of infection quickly (Radostits
et al., 2000), while adequately fed animals are better able to tolerate parasitism (Urquhart et
al., 1996). Protein supplementation to the young grazing sheep has reduced the need for
survival drenching and increased the production (Van Houtert et al., 1996).
In tropical Africa where small ruminants are kept under grazing management, the feed
availability and quality mainly depends on the season of the year. Wet season being good
both for feed availability and quality is also conducive for parasite larvae survival and
development on pasture, while dry season is bad both to the host and parasite larvae. But
during dry season, accumulation of flocks along water courses for the search of water and
green grass favours disease transmission and spread between flocks (Troncy, 1989; Teklye,
1991; Radostits et al., 2000).
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2.3.1.2 Grazing behaviour of hosts
Grazing behaviour of the host is one of the influencing factors affecting the epidemiology of
gastrointestinal tract nematodes. Sheep grazing close to ground are more exposed to massive
numbers of infective larvae compared to free ranging goats that are less exposed to the
infective larvae since their feeding behaviour includes a large component of browsing at levels
well above the ground (Smith and Sherman, 1994). However, domesticated goats managed in
areas where access to browse is restricted and pasture grazing is mandatory are equally or
even have greater risk of nematode parasitism (Smith and Sherman, 1994; Radostits et al.,
2000).
Overstocking and prolonged grazing on the same pasture leads to overgrazing (loss of
available herbage per animal) and hence lower plane of nutrition. It also encourages large
amount of faecal deposit on the grazing field and potentially to higher level of infectivity per
unit area. Thus overstocking and prolonged grazing on a plot of land, besides affecting the
growth rate of animals, it may exacerbate the pathogenic effect of acquired infections by
lowering the protective immunity (Dunn, 1978; Troncy, 1989; Ndamukong et al., 1994;
Thamsborg et al., 1996; Urquhart et al., 1996).
Rotational grazing, alternate grazing, mixed grazing and zero grazing having knowledge on
the epidemiology of a given nematode parasite, are some of important management techniques
to control the nematode parasite with out chemical use (Dunn, 1978; Kaufman, 1996). But the
use of rotational grazing to control ruminant GIT nematodiosis was said of little value because
the infective larvae may resist long period on the grazing field (Soulsby, 1982; Radostits et al.,
2000).
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2.3.2 Environment-Parasite Interactions
Fourth and fifth stage larvae and adult gastrointestinal tract nematodes of sheep and goats
reside in the GIT, while eggs, first stage; second stage and third stage larvae exist on the
external environment (Dunn, 1978). Having conducive temperature and moisture, nematode
eggs deposited by carrier sheep and goats are able to develop to the third stage larvae (L3) in
the external environment. But external factors do influence the development from egg to
infective larvae. These external factors to be included in environment-parasite interactions and
thus affecting the epidemiology of GIT nematodes are mentioned as follows:
In cool temperate climate free-living stage of parasite larvae could survive for longer period
than arid tropical environment (Dunn, 1978; Chiejina et al., 1989). In the cool tropical
environment of Ethiopia, Tembely et al., (1997) reported large number of nematode parasites
from small ruminants during wet season. Similarly peak infection rate of gastrointestinal tract
nematodes were reported during the rainy season and lower infection rate during dry season of
the year in Southern Nigeria (Fritsche et al., 1993) and in The Gambia (Anene et al., 1994).
This seasonal fluctuation may be due to a number of factors which are responsible for the
numbers and availability of infective stages, and these may be conveniently be grouped as
factors affecting contamination of the environment, and those controlling the development and
survival of the free living stages of parasites (Anene, 1994; Urquhart et al., 1996; Stromberg,
1997; Craig, 1998).
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2.3.2.2 Biotic potential and survival capacity
Some nematode eggs survive outside of their host for considerable period of time (ex.
Trichuris, Ascaris, Nematodirus), which may be dependable on the thickness of their egg shell
thus responsible for longer time contamination of the grazing field (Anene et al., 1994).
2.3.2.3 Desiccation
Temperature and humidity are major determining factors among others for the development
and survival of nematode egg and free-living stage of larva. Under favourable condition, the
egg will be hatched to first stage larvae with in few hours after its passage through faeces.
The consecutive first and second stage larvae are developed, and these egg and larvae are
extremely susceptible to the desiccation (Craig, 1998). A study conducted in Awassa, southern
Ethiopia, has indicated that there was significant difference in temperature and moisture
content of sun exposed and under shade faecal masses (Demelash et al., 2004). Higher
temperature and low moisture were observed in sun exposed and lower in under shade. And
also egg counts and larvae counts between sun exposed and under shade faecal masses was
revealed that higher egg and larvae counts were encountered in those under shade and lower
observed in sun exposed (Demelash et al., 2004). But embryonated eggs of some nematode
species and ensheathed third stage larvae are best equipped to survive the adverse conditions
of the environment (Anene et al., 1994). However, both direct sunlight and prolonged high
ground temperature are fatal and hence no stronglylid life cycle can be completed in totally
arid environment and desert region (Chiejina et al., 1989; Bowman, 1999) unless
microhabitats that provide enough moisture exist.
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2.3.3 Host-Parasite Interaction
Analysis of nematode population in ruminants over a period of several years unveiled the
presence of host and worm related factors affecting the survival of larval nematode inside a
host (Craig, 1998).
Generally, the effect of a protective host immune response is to limit the parasite population
with in certain limits acceptable to the well being of the host. This action is achieved by:
elimination or expulsion of parasites; retardation or inhibition of development of newly
acquired infection; reduction in the fecundity of female worms and reduction in the
establishment rate of infections (Kelly, 1973). The defence status of a host naturally vary
between species of host (sheep versus goats) and between individuals in a flock (Stear et al.,
1995; Radostits et al., 2000); however, in many host parasite relationships where strong
immunity develops, this is usually associated with previous exposure to the parasite (Bowman,
1999). Fluctuations in the immune status of grazing ruminants can also happen due to
concurrent diseases, stress, steroid therapy and other physiologically related factors. Most
commonly mentioned factors include:
This phenomenon is observed in pregnant and lactating ewes at the time of two weeks before
lambing to four weeks post parturition (Dunn, 1978; Urquhart et al., 1996; Bowman, 1999).
The responsible factor for this rise of faecal egg count is said to be the release of lactogenic
hormone, prolactine (Kelly, 1973; Urquhart et al., 1996; Bowman, 1999) which suppress the
immunity of the ewe and brings an apparent increase of the numbers of worms by the
resumption of development of previously inhibited larval stages, increased rate of
establishment of newly acquired larvae, failure in the elimination of existing infections,
uninhibited development to maturity of newly acquired larvae, increase in the fecundity of egg
13
laying adult female nematodes, which were repressed in ordinary healthy non-lactating ewes
(Kelly, 1973; Dunn, 1978). The condition in general brings increased worm egg out put which
is epidemiologically significant not only by contaminating the pasture but also its adjustment
with the period of existence of new susceptible population of lambs (Kelly, 1973; Urquhart et
al., 1996).
In endemic area, after heavy rainfall; the faecal egg out put of certain nematodes
(Haemonchus contortus) drop sharply to near zero level due to expulsion of adult worm
burden. This situation is related to ingestion of massive third stage larvae that trigger an
immediate type of hypersensitivity reaction due to ingested antigen (Soulsby, 1982; Urquhart
et al., 1996). Another explanation suggested was that self cure is due to ingestion of fresh
growing grass that induces diarrhoea and thus resulting in expulsion of adult parasites. But
whatever the cause, self-cure is probably of mutual benefit to both host and parasite. The host
gets temporary relief from persistent blood loss while the aging parasite population is
eventually replaced by a vigorous young generation (Urquhart et al., 1996).
The existence of age related difference in susceptibility/resistance of sheep and goat flocks to
nematode infections has been reported by several researchers. Soulsby, (1982) has indicated
that young sheep and goat are susceptible to Trichostrongylus species than aged (older ones).
Sheep aged 16 or 28 months showed better immunological response to nematode infection and
suppressed nematode egg count in their faeces than 4-month-old lambs (Douch and Morum,
1993). Others relate age related susceptibility to various stresses they face, such as weaning
stress and decreased milk production in ewes due to various reasons and hence reduced milk
intake by suckling lambs and kids (Troncy, 1989; Silva et al., 1998). Apparently, the immune
response controlling the parasite populations in the host is readily impaired or deficient in
young animals and, these are particularly susceptible to the pathogenic effects of infection
(Kelly, 1973), which is best indicated by the persistent of infection by young animals is
14
reflected by the continued passage of worm egg in their faeces for up to 60 days after infection
(Kelly, 1973). However, Fritsche et al., (1993) in their work have suggested that acquired
immunity to be the major factor for susceptibility or resistance to nematode infection.
Sheep industries through out the world are seeking alternative approaches to parasite control
because of production losses due to parasitism, increased frequency of drug resistance
parasites and consumers demand for minimal chemical use in sheep and goat production
(Owen & Axford, 1991; Kloosterman et al., 1992). Variation in susceptibility between breeds
and with in breeds due to genetic factor were reported by different researchers in the world
(Soulsby, 1982; Owen and Axford, 1991; Gray et al., 1992; Hohenhaus et al., 1995; Gray,
1997; Amarante et al., 1999). Research in sub humid coastal Kenya has clearly shown that
indigenous Red Maasai sheep are more resistant to gastrointestinal nematodes (predominantly
Haemonchus contortus) than exotic Droper sheep (cited by Woolaston, 1996). The study
conducted on lambs of Suffolk and Gulf Coast Natives for naturally acquired strongyle
infection, have shown that native lambs of the Gulf Coast developed resistance to
Haemonchus contortus infection during their first exposure to infection at the age when they
are considered immune incompetent and colostrally transferred anti Haemonchus contortus
immunoglobulin did not appear to be involved in the resistance (Bahirathan et al., 1996).
Lines of Romany sheep, selected as lambs from consistently low or high faecal egg count
(FEC) parents following periods of natural challenge in New Zealand, indicated significant
difference between breeding lines for abomasal and intestinal nematode resistance (Bisset et
al., 1996). These lambs of resistant genotype had lower faecal worm egg counts, lower worm
burden and higher level of resistance to larval establishment (Gray et al., 1992). This genetic
difference is moderately heritable, and could be effectively selected by using faecal worm egg
count although many alternatives, such as DNA markers, host antibody and parasite antigen
assays are being developed for use as selection criteria (Gray, 1997) and these genetically
resistant individuals can be used for disease control (Barger, 1989; Stear and Murray, 1994).
15
2.3.3.4 Arrested development of the parasite in the host (Hypobiosis)
The major factors that initiate hypobiosis are said to be acquired immunity of the host
(pervious exposure) or the experience of climatic conditions during the free-living phase. In
the tropical environment, reduced moisture and increased temperature are claimed to induce
hypobiosis (Eysker, 1997; Gatongi et al., 1998). The hypobiotic larva is not only able to evade
unfavourable conditions but also evade the host’s immunological surveillance system by being
metabolically inactive (Fritsche et al., 1992; Eysker, 1997; Gatongi et al., 1998; Craig, 1998;
Tembely et al., 1997; Tembely, 1998).
The presence of strain differences in parasite species was stated in Urquhart (Urquhart et al.,
1996). The experiment done in two strains of Haemonchus contortus; Sheep Derived Strain
(SDS) and Goat Derived Strain (GDS) indicated that, GDS appeared to affect goats more
severely than SDS even though there was no difference in their establishment (Rahman and
Collins, 1991).
16
2.4 Diagnosis
The problem of gastrointestinal tract nematodiosis is one of the major problems of small
ruminant production and health. Due to this great impact imposed, early detection and
correction is of paramount importance. Clinical diagnosis of gastrointestinal tract nematodes
of sheep and goats needs history of the area, history of anthelmenthics treatment, grazing
history, age of animal and clinical signs manifested by the disease (Troncy, 1989), but as GIT
nematodiosis share common clinical manifestations with other diseases laboratory diagnosis is
important.
Although there is much current interest in the use of serology as an aid to the diagnosis of
helminthosis, particularly with the introduction of enzyme linked immuno sorbent assay
(ELISA) test, diagnosis of gastrointestinal parasitic infections still depends mostly on
parasitological findings of eggs and/or parasites in faecal samples (Urquhart et al., 1996).
Faecal examination for the detection of worm eggs is most common and routine work in
gastrointestinal tract nematode diagnosis (Urquhart et al., 1996). Examination of faeces for
nematode eggs may vary from a simple direct smear to more complex methods involving
centrifugation and the use of floatation fluids (Hendrix, 1998). Various techniques were
developed and used, but some are summarized as follows:
17
2.4.1.1.1 Qualitative methods
The presence or absence of worm eggs in faecal sample by the use of direct smear of fresh
faeces on a microscope slide and examination under low power objectives of the microscope is
routine procedure. However, this technique is only useful to detect nematode eggs when it
exists in high concentration in the faeces. Other disadvantages of direct techniques include
difficulty to identify them since the eggs are partially covered by debris materials and
quantitative results could not be obtained although it is fast and easy technique (Hendrix,
1998).
Light infections are not easily detected by the use of direct smear; therefore concentration
technique was developed to over come the shortcomings of direct smear. The concentration
techniques that are widely used include; the use of salt or sugar solution and centrifugal
concentration techniques. In both cases the logic behind is to concentrate the nematode eggs in
a given portion of sample or processed faecal material (Hendrix, 1998; Bowman, 1999). In
flotation the type of egg recovered is related to specific gravity of solutions; half saturated
sodium chloride (NaCl) with specific gravity of 1.125 is capable of floating trichostrongyloid
and strongylid eggs while fully saturated sodium chloride solution with specific gravity of
1.204 is preferred as general purpose solution (Hendrix, 1998).
18
difficult to relate directly with the burden of parasites in large ruminants (Hendrix, 1998), still
it is widely used and best correlation was observed in small ruminants (Gray, 1997). And the
method is also used to detect anthelmenthic resistance and to distinguish between susceptible
and resistant breeds for genetic selection (Eysker and Ploeger, 2000).
Grazing sheep and goats usually have mixed nematode infections. Only few nematode
parasites have characteristic eggs that enable us to differentiate them to genus level
(Nematodirus spp., Trichuris spp., Strongyloides spp.), but those of tichostrongyle and
strongyles are not easily differentiated, for this reason faecal culturing and larval identification
based on the keys available is useful technique (Soulsby, 1982; Hendrix, 1998).
Representative animals are used (selected) and followed for longer period of time. This
method is used to follow the dynamics of nematode population. In each selected period of time
animals are necropsied and worm burden is counted (Eysker and Ploeger, 2000). Post-mortem
examination of gastrointestinal tract for adult worm is a definitive diagnosis; worms recovered
from specific sites could be identified by the use of morphological features based on keys
(Hendrix, 1998; Urquhart et al., 1996).
19
3. MATERIALS AND METHODS
The study was conducted in three selected agro-ecological zones of Southern Ethiopia
(Southern Nations, Nationalities and Peoples’ Regional State). The selected agro-ecological
zones of the study sites were Bonke (highland agro-ecology) and Mirab-Abaya (lowland agro-
ecology) from Gamo Gofa zone and Soddo-Zurya (midland agro-ecology) from Wolayta zone
based on the altitude and weather condition.
1. Bonke (Highland)
Bonke is one of the 11 weredas of Gamo-Gofa zone, situated at 554 kilometres southwest to
Addis Ababa. The wereda has three agro-ecological zones of which 46% is highland (>2400
masl), 24% midland (1600-2400 masl.) and 30% is lowland (600-1600 m.a.s.l.). The altitude
range of the wereda is 600 to 4207 masl (top of mount Guge). The topography of the area is
18% is mountainous, 64%is sloppy and 18% is plain land. 10.8% of it was cultivated, 21.3%
covered by forests and bushes, 10.8% was allocated for grazing and the rest 34.2% is for other
purposes (Gamo-Gofa zone BRD, 2002/03).
The area receives mean maximum rainfall of 2200 mm and the mean minimum was 600 mm,
with the mean average rainfall of 1500mm annually. The mean maximum temperature of the
area was 300c and the mean minimum 40c and the average of 160c. In this wereda the study
was based on the altitude range of 2430 to 3030 m.a.s.l. (Gamo-Gofa zone BRD, 2002/03).
2. Soddo-Zurya (Midland)
Soddo-Zurya is one of the seven weredas of Wolayta zone and located at about 390 kilometres
southwest to Addis Ababa. The wereda has three different agro-ecological zones; 6.13%
highland, 87.7% midland and 8.4% is lowland. The altitude range of the area is 1200 to 2950
m.a.s.l. (top of mount Damota). The area receives total annual rainfall of 1112.3 mm and the
20
annual average maximum temperature of 25.0 0c and annual average minimum temperature of
14.50c(W Z R D D, 2003).
3. Mirab-Abaya (lowland)
Mirab-Abaya wereda is one of the weredas of Gamo-Gofa zone located at about 465
kilometres south west of Addis Ababa. The total area of the wereda is 97, 975 hectare and
66.7% of it is lowland (1170 to 1600 masl), 10.5% is midland (1600 to 2400) and 21% is
highland (>2400masl). Of the total land area, 55% is plain land, 15% is sloppy, 19%
mountainous and 11% is valley. The annual rainfall of the area was 500 to 580 mm in lowland
and 1000 to 1100 mm in highlands (Gamo-Gofa zone BRD, 2002/03).
The vegetation of the study areas is based on the respective agro-ecological zones. In the
highlands coniferous trees, eucalyptus, bamboo plantation and intensively cropped fields with
enset and other crops. The lowland part of the study areas are characterized by acacia trees,
gully forests, bushes and grass lands. Various wild fauna are known to be hosted in these study
areas, among which, lion, leopard, hyena, foxes, antelopes, large wild ruminants such as
gazelle and buffaloes and zebras are predominantly exist.
21
Figure 1. Map of Ethiopia, SNNPRS and the Study areas
22
Live stock population
In all study areas the predominant species of the livestock are cattle, sheep and goats (CSA,
2003). The details of livestock population of study zones and study weredas are indicated in
tables 1 and 2 below.
In all agro-ecological zones of the study areas sheep and goats were housed in rainproof
houses either separately or together with large ruminants and human being. The livestock
management of the different agro-ecological zones of the study areas have more or less based
on grazing all year round on natural pastures. In all areas, communal type of grazing system
predominates with mixed livestock grazing in the state owned grazing fields but occasionally
sheep and goats were found tethered on pegs and trees.The goats of the lowland has free
23
access for browsing in wider permanent grazing lands (Annex 14, plates 1 & 2) that has
bushes and forests while those of midlands and highlands were forced to graze together with
other livestock classes (Annex 14, plates 4 - 9). Supplementation with crop residues, grass,
and household left over, grains and root crops was common practice in midlands and
highlands while occasional in lowlands during the night.
In all cases there are well-equipped veterinary clinics in all weredas of the study area at the
centre. The health problems of small ruminants that are frequently mentioned include:
gastrointestinal tract helminth and lung worms in highlands, gastrointestinal tract helminth and
ecto parasites in midlands and CCPP, mange mites and pasteurelosis in lowlands (GGZRDD,
2003; WZRDD, 2003).
The study animals are sheep and goat population raised in the highland areas of Bonke,
midland areas of Soddo-Zurya, and lowland areas of the Mirab-Abaya. According to Bureau
of Rural Development of Gamo-Gofa and Wolayta, the estimated figures are indicated in the
table 3 below.
24
Table 4. The age structure of the study animals
The sample sizes for the post mortem and coprological examination of the study were as
follows.
Post mortem examination for nematode species identification and worm burden determination
was carried out starting from September 2004 up to April 2005. Samples were obtained by
systematic random sampling method from sheep and goats slaughtered in three towns that
belong to different agro-ecological zones. 12 sheep and goats were sampled per season per site
from midland and lowland agro-ecological zone and 15 sheep and goats were sampled per
season in highlands. A total of 180 GIT samples were obtained during the study period; 61
from highland, 60 from midland and 59 from lowland agro-ecological zones of which 101
were goats and 79 were sheep. 172 were obtained from local hotels and 8 were purchased.
For each wereda and both species of animals the sample size was determined by the use of the
formula in Thrusfield, (1995) as indicated below:
25
N = 1.962 x PQ/D2
Where N is required sample size, P is expected prevalence based on previous preliminary
surveys, Q is 1-P and D is the level of precision (5%), 1.96 to indicate 95% confidence level.
The study period was divided into four based on the seasonal condition of the area as late wet
season (August and September), early dry (November and December), late dry (February and
March) and early wet (April and May). Faecal samples for prevalence and epg determination
were collected during these four seasons.
Therefore based on the formula our sample size for each wereda was 113 sheep and 138 goats
by considering 92.0% prevalence of GIT nematodiosis in sheep population and 90.0%
prevalence in goat population from previous surveys made in the surrounding areas (Dereje,
1992; Haileleul, 2002). This makes a total of 339 sheep and 414 goats, and a total of 753
faecal samples from three agro-ecological zones at a season but due to little goat population at
high lands only 80 to 90 animals were sampled at a time and a total of 2,828 faecal samples
were collected by systematic random sampling for prevalence determination and epg counting.
The samples of faeces for egg counting (epg) and worms recovered from GIT were preserved
in 10% formaline and 70% alcohol, respectively for later examination in the laboratory.
Cross sectional study type for prevalence determination by post mortem and faecal
examination and prospective longitudinal study type for studying the seasonal dynamics of
nematodes was employed (Toma et al., 1996)
A day before slaughtering, ante mortem investigation was performed and properly recorded
for each animal concerning its village of origin, age, sex, body condition score and its general
health condition (Annex 1 and 2).
26
The body condition and age of the animals were determined using the body condition scoring
and age determination method developed for respective species by Gatenby, 1991 and Mike,
1996(Annex 8 and 9).
Following slaughter the gastrointestinal tract was removed, and the abomasum, small intestine
and large intestine were immediately isolated by three ligatures (between omasum and
abomasum, abomasum and small intestine, ileum and caecum) to avoid mixing of the contents.
Collection of the contents of abomasum and intestines, and recovery of nematode species was
according to, MAFF, (1977) and Urquhart et al. (1996). Identification and counting
procedures was done according to MAFF, 1977; Soulsby, 1982; Jorgen, H. and Brian, P. 1994,
Kaufmann, 1996 and Urquhart et al., 1996(Annex 10, 11, 12 )
In a mixed infection by nematode species, the intensity of nematode count was classified as
low (< 2000 nematode), moderate (2000-10,000) and high (>10,000) as described in Radostits
et al., (2000).
B. Coprological method
Faecal samples obtained from field (population) and from slaughtered animals were subjected
to qualitative (flotation) and quantitative (modified McMaster) techniques (Annex 13). Faecal
egg count was determined using modified McMaster technique, each nematode egg counted
represents 50 eggs per gram of faeces, when the faecal samples became negative for nematode
egg in modified McMaster technique, it was subjected to flotation technique for prevalence
determination.
Classification of the intensity of infection was made based on faecal egg counts as light (50-
800 epg), moderate (801-1200 epg) and heavy infections (>1200 epg) as described in Jorgen,
H. and Brian, P. (1994) for the mixed infections in grazing small ruminants.
27
3.1.1 Statistical analysis methods
The data collected was recorded in Microsoft excel. The independent variables analysed were:
agro-ecological zones (highland, midland and lowland), species of animals (Goat and sheep),
sex of animals (male and female), age of animals (less than 1 year, 1-2 years, more than 2
years), body condition of animals (very thin, thin, moderate, fat) and season of the year (late
wet, early dry, mid dry, late dry, early wet) but in the case of faecal samples obtained from the
field mid dry was not used. The dependant variables analysed were faecal egg counts and
nematode counts. Because of the skew ness in their distribution, faecal egg counts and
nematode counts were subjected to a logarithmic transformation [log (x+1)] for analysis.
To compare the difference between means and proportions two soft wares were used.
STATA 7.0 was used for univariate analysis of variance (ANOVA) to compare more than two
means, t-test to compare the difference between two means, and chi-square to compare the
difference between the proportions for faecal egg count (epg) and post mortem (nematode
burden) results. SPSS version 11.5.0 was used to calculate means for egg counts and nematode
counts and to detect significant differences between and with in subject effects and to correlate
epg count to nematode burden of slaughtered animals.
Mean nematode egg counts and mean nematode burden were calculated and presented by
using geometric means (antilogarithm).
28
4. RESULTS OF THE STUDY
A total of ten different species of nematodes were recovered from the gastrointestinal tracts of
79 sheep and 101 goats during the study period (Table 5). The parasite species identified and
their percent prevalence in decreasing order was as follows: Oe. columbianum, 92.2; T.
colubriformis, 73.9; H. contortus, 68.9; T. axei, 61.7; T. ovis, 42.8; B. trigonocephalum, 18.9;
Teladorsagia spp.,15.6; T. probolurus, 5.0; S. papillosus, 1.1 and C. curticei, 0.5. The
Teladorsagia spp. and C. curticei were only encountered in the highland agro-ecological zones
of the study site where as Strongyloides papillosus and Trichostrongylus probolurus were
encountered in midlands and lowlands.
The over all prevalence of GIT nematodiosis of small ruminants in the three agro-ecological
zones of the study sites was 98.9% and in the agro-ecological bases; 98.3, 98.3 and 100.0% in
lowlands, midlands and highlands, respectively. There was significant individual nematode
species prevalence variations (p<0.05) among different agro-ecological sites except for Oe.
columbianum and S. papillosus as indicated in the table 5 below.
4.1.2 Post mortem findings based on season, agro-ecology, age, species & sex differences.
29
Table 5. Prevalence rates of GIT nematode species by agro-ecological zones
30
Table 6. Percent prevalence of GIT nematode species of small ruminant on age base
The identified nematode species from sheep and goats slaughtered during different seasons of
the year (41in late wet, 40 in early dry, 39 during mid dry, 42 in late dry and 16 during early
wet) have indicated significant seasonal variation. The seasonal point prevalence of H.
contortus, T. axei, B. trigonocephalum and Oe. columbianum were varied highly significantly
(p<0.001) between seasons of the year; while those of T. ovis and T. colubriformis point
prevalence were significantly varied (p< 0.05) between different seasons but the over all point
prevalence change for total nematode species was not significantly different between seasons
of the year(p>0.05). The seasonal point prevalence changes of six nematode species is
indicated in the figures 2, 3 and 4 below.
31
Figure 2. Oe. columbianum and T. axei seasonal point
prevalence
120
97.6
100
92.3
90.2
85.7
100
76.2
LW
69.2
75
80
ED
61
60 MD
35.7
40 LD
EW
20
0
Oe. columbianum T. axei
LW = Late wet season, ED = Early dry, MD = Mid dry, LD = Late dry, EW = Early wet
60
46.3
50
38.5
38.1
LW
40
31.3
ED
23.8
30 MD
20.5
19
LD
14.6
12.5
20
EW
10
0
B. trgonocephalum T.ovis
32
Figure 4. T. colubriformis and H. contortus seasonal point prevalence.
87.5
87.5
100
76.9
75.6
90
73.8
71.4
69.2
66.7
80
63.4
61.9
70 LW
60 ED
50 MD
40 LD
30
EW
20
10
0
T.colubriformis H. contortus
The total prevalence of GIT nematodiosis of small ruminants in this study was not
significantly different between agro-ecology, between seasons of the year, between species of
animals and between male and female. Significant differences were observed for mean total
nematode burden and mean total epg count from slaughtered animals during different seasons
of the year (p<0.05). There was no significant difference in the mean total worm burden and
mean total epg count for sex difference of slaughtered animals (p> 0.05 in both cases) &
species variation did not revealed significant difference in mean total nematode burden. The
details of findings are indicated in the table 7 below.
33
Table 7. Prevalence, MTNB and MTEPG count by agro-ecology, season, species and sex of animals.
PM = Post mortem, MTNB = mean total nematode burden, SE = standard error, MTEPG = mean total egg per gram of faeces,
HL = highland, ML = midland, LL = lowland, LW = late wet, ED = early dry, MD = mid dry, LD = late dry, EW = early wet
34
The results of mean nematode burden of different seasons of the year in three different agro
ecological zones showed significant differences both for seasonal and agroecological variation
(p<0.05). These significant variations were observed between seasons of late wet and early dry
(p<0.05), late wet and mid dry (p<0.05), late wet and late dry (p<0.05), late wet and early wet
(p<0.05) and between agro-ecological zone of highland and lowland during late wet season
(p<0.05). The mean nematode burden in seasons of the year of three different agro-ecological
zones is indicated in the table 8 below.
Table 8. Mean GIT nematode burden from sheep and goats by agro-ecology and season
The results of necropsied animals has shown that 58.4% of the infected animals harboured
light infestation, 40.5% were moderate infection and only 1.1% were severely infected by
gastrointestinal tract nematode species of small ruminants during the study period. The
findings are presented by species of animals in the table 9 below.
Table 9. Intensity of nematode count from sheep and goats in the study area
35
The results of post mortem indicated that out of 98.9% of infected animals in the study area,
97.7% were infected by more than one nematode species. On agro-ecological bases; 96.6%,
98.3% and 98.4% of infected sheep and goats in midland, lowland and highland agro-
ecological zones of the study sites respectively had mixed nematode infections and only 3.4%,
1.7% and 1.6% of infected individuals in respective agro-ecological areas harboured one
nematode parasite species. The number and percentage of poly parasitised animals in different
study sites are indicated in the table 10 below.
Table 10. Number and percentage of nematode species infecting parasitised sheep and goats
Poly parasitism of more than three nematode species in a single host predominates in all agro
ecological areas and varies from 57.4% in highland to 74.2% in lowland. A total of 64% of
poly-parasitised animals in all agro-ecological zones were infected by 4 or more nematode
species.
36
4.1.3 Correlation of total nematode count to total epg counts from slaughtered sheep and
goats.
The mean total nematode count and mean total epg count both from slaughtered animals in
this study was positively correlated, and their correlation was significant (Spearman’s rho
(rs)=0.57, p=0.000). The correlation of mean epg to individual nematode species burden in this
study was found to be significant for three nematode species; T. colubriformis, rs=0.50; H.
contortus, rs=0.45 and Oe. columbianum, rs=0.39 with p=0.000 for all correlations. The
correlation of mean total nematode burden to mean total epg is indicated in the figure 5 below.
Figure 5. MTNC & MTEPG both from slaughtered sheep & goats in different seasons.
4
3.5
3
Log(X+1)
2.5
MTEPG
2
MTNC
1.5
1
0.5
0
Late wet Early dry Mid dry Late dry Early wet
37
4.2 Coprological results
Faecal examination results in all study sites from population of sheep and goats during four
sampling periods; late wet (August), early dry (November), late dry (February) and early wet
(April) indicated significant seasonal variation (p<0.05) for all study sites. The coprological
point prevalence and their 95% confidence interval indicated the significant difference in the
prevalence of GIT nematodiosis between sheep and goats during late wet season in lowland
and highland; and late dry and early wet seasons in highland. In all seasons of the year the
point prevalence of GIT nematodiosis from coprology in highland indicated the higher
prevalence in goats than in sheep. Detail findings of coprological examination are indicated in
the figures 6, 7 and 8 below.
Figure 6. Seasonal coprological point prevalence(%) of GIT nematode infection of sheep (s)
and goats (g) by agro-ecology.
120
100
80
s
60
g
40
20
0
Late Early Late Early Late Early Late Early Late Early Late Early
wet dry dry wet wet dry dry wet wet dry dry wet
The coprological examinations of sheep and goats in the total study sites during different
seasons of the year indicated the highest prevalence during early dry period than other seasons
of the year. Significant difference in the point prevalence between sheep and goats was
observed during late wet and early wet seasons of the year.
38
Figure 7. Seasonal coprological point prevalence (%) of GIT nematode infection of sheep (s)
and goats (g) in all study sites
90
80
70
60
50 s
40 g
30
20
10
0
Late wet Early dry Late dry Early wet
Figure 8. Seasonal point prevalence percentage of GIT nematode infection of small ruminants
in all study area by coproscopic examination.
100
80
60
40
20
LW ED LD EW
39
4.2.2 Results of faecal egg count
The majority of infected animals had a faecal egg count in the range of 50-800 epg and only
few proportions of animals had faecal egg count over 1200. The results are indicated in the
Figure 9 below.
100%
80%
>1200
60% 801-1200
40% 50-800
0
20%
0%
Late wet Early dry Late dry Early wet
0= indicating zero faecal egg count (no nematode egg counted per gram of faeces)
50-800= 50 to 800 epg of faeces (light infection),
801-1200= 801 to 1200 epg of faeces (moderate infection) and
>1200= more than 1200 epg of faeces, indicating heavy infection with GIT nematodes.
4.2.2.1 Fecal egg counts based on season, agro-ecology, age, species and sex differences.
Mean epg count in agro-ecological zones have showed highly significant variation in different
season of the year (p<0.05, p< 0.05 and p< 0.001 for early dry, late dry and early wet seasons
respectively), but during late wet season mean epg counts of different agro-ecological zones
was not significantly different (p>0.05).
Age differences of the animals had not shown significant difference in epg count in all seasons
of the year (p>0.05). A species difference has revealed significant differences only during late
wet and early wet seasons of the year (p< 0.05). Difference in body condition of sampled
40
animals has shown significant differences only during early wet season of the year (p<0.05).
Detail results are presented in the table 11 below.
41
Table 11. Mean epg count by agro-ecology, age group, body condition, animal species and season.
42
5. DISCUSSION
The post mortem examination of 180 sheep and goats during the study period enabled the
identification of ten (10) nematode species parasitising the small ruminants in three agro-
ecological zones. The existence of nematode species such as Oe. columbianum, T.
colubriformis, H. contortus and T. axei in higher percentage through out the study period in all
agro-ecological zones of the study sites agrees to the findings of other workers in other parts
of the country (Bayou, 1992; Dereje, 1992; Esayas, 1999; Kasambara, 1999). But the
identification of Teladorsagia spp. and C. curticei from highlands and T. probolurus from
midland and lowlands in this study was new to the region and even to the country after Graber,
(1975) in which he reported Teladorsagia spp. from sheep at Debre-Berhan area and T.
probolurus from camels in Harrar.
The prevalence of GIT nematodiosis of small ruminants from necropsied animals indicated
98.9% (97.5% in sheep and 100.0%in goats). This high prevalence of GIT nematodiosis of the
present study agrees with previous findings in different corners of the country and other parts
of the tropical countries (Gebreyesus, 1986; Esayas, 1988; Tesfa-alem, 1989; Melkamu, 1991;
Dereje, 1992; Genene, 1994; Bonfoh et al., 1995(in Togo); Esayas 1999; but higher than the
reports of Jacquiet et al., 1995 (in Mauritania); Abdala and Elmalik, 1997(in Sudan); Achnef,
1997; Berrag and Cabaret, 1998 (in Morocco) and Haileleul, 2002(in southern Ethiopia)).
The moderate prevalence rate of Teladorsagia spp. in highland (45.9%) is indicative of its
economic and pathogenic importance to small ruminant production of the area since it is
pathogenic especially during its immature stage (Dunn, 1978; Jorgen and Brian, 1994;
Urquhart et al., 1996). The higher prevalence rate of Oe. columbianum (90.0% in midland,
91.8% in highland and 94.9% in lowland) agrees with the previous findings of Dereje (1992);
Kasambara (1999) but higher than the findings of Ahmed (1988); Esayas (1988); Tesfa-alem
43
(1989); Melkamu, (1991); Bayou (1992); Yoseph (1993); Getachew (1998); Abebe and
Esayas (2001); Haile-leul (2002) in different parts of Ethiopia and Bonfoh et al. (1995) in
Togo; Nwosu et al. (1996) in Nigeria. Oe. columbianum is known for its ability to form
numerous nodules in small and large intestines of previously exposed animals thus causing
enormous economic loss by damaging the mucosa of intestines and hampering the digestive
and absorptive function of gastrointestinal tract of infected sheep and goats (Graber, 1975;
Soulsby, 1982; Smith and Sherman, 1994; Urquhart et al., 1996).
The magnitude and distribution of some of the identified nematode species seems to be related
with the agro-ecological zone of the study area: Teladorsagia spp. was identified only in the
highland from sheep and Haemonchus contortus prevalence was significantly higher in
lowlands than highlands (p<0.001). This difference in their prevalence might be related to
their geographical origin; since Teladorsagia is a temperate parasite and therefore it is adapted
to cool environment, while Haemonchus as tropical or sub-tropical parasite is well adapted to
warm climate (Craig, 1998; Radostits et al., 2000).
The findings of the overall high nematode prevalence together with high nematode burden in
all study sites is suggestive of the importance of the infection in small ruminants all over the
region, irrespective of the variations in climate and vegetations that exist, and this finding
differs from the work of Anene et al. (1994) in Southern Nigeria, who reported significant
variations in prevalence and worm load in various agro-ecological zones.
44
The finding of significantly higher nematode prevalence by coprological examination in goats
than sheep in highlands during most of the seasons of the study period agrees with the finding
of Jacquiet et al. (1992) in south-west Mauritania and this might be related to the more
susceptibility of goats than sheep when they are equally challenged (Smith, 1994).
The coproscopic finding of a significant increase (p<0.05) in early dry season prevalence of
nematodes (77.4%) compared with 57.5% prevalence in late wet season is in agreement with
the results of Yoseph (1993) and Achenef (1997). The increase in coprological point
prevalence during the early dry season might be related to the lowered host immunity because
of the decline in feed quality and quantity as dry season starts. As the temperature and
moisture during such period are, however, conducive to free living stages of parasite, there
will be increased establishment with continued challenge and increased fecundity of the
existing adult nematodes (Troncy, 1989; Agyei, 1996).
The prevalence of Oe. columbianum and Trichostrongylus spp. infestation has shown
increased seasonal pattern, September through April, while those of Trichuris ovis and B.
trigonocephalum were decreasing starting from September to April. An increasing trend in
prevalence of Trichostrongylus spp. in relation to season was observed by Bekele et al.,
(1982). This variation in prevalence with changes in the season might be related to nematodes
specific behaviour. The release of inhibited larvae from nodules during dry season when there
was feed shortage (stress) in case of Oe. columbianum and the resistance of Trichostrongylus
spp. larvae to desiccation of dry season than others might favoured its increasing trend (Dunn,
45
1978; Troncy, 1989). The decreasing pattern in point prevalence of B. trigonocephalum might
be due to the unfavourable condition of dry season for skin penetration and that of Trichuris
ovis might be due to the ingestion of the infective larvae within the eggshell by the host
usually at the end of dry season, when the grass biomass diminishes and the animals are forced
to graze near to the ground (Graber, 1975; Radostits et al., 2000).
The increased mean nematode burden obtained during the early wet season (1927.12359.9)
compared with late dry season (1348.0334.3) is in agreement with other findings in different
parts of the world ( Agyei, 1996; Nwosu et al., 1996; Abdalla and Elmalik, 1997; Egbe, 1999;
Nginyi et al., 2001; Etana, 2002 ) and is probably attributed to the highest number of infective
larvae present on the pasture at early wet season and exposure to a high larval challenge in an
already weakened animal due to previous drought (Troncy, 1989).
The increment in mean nematode burden recorded during mid dry season than early dry
season, when the weather was harsh to free living stage of nematodes, might be due to the
previously established larvae in the host during early dry season and the reduced efficiency of
the host immune system due to decreased quality and quantity of feed (Troncy, 1989;
Urquhart et al., 1996).
The nematode egg out put (epg) variation obtained during the different seasons of the year was
significant particularly between late wet and late dry (p<0.05); between late wet and early wet
(p<0.05); between early dry & late dry (p<0.01) and between early dry & early wet (p<0.05).
This finding of present work agrees with the results of Jacquiet et al. (1995), Fritsche et al.
(1993), Assefa & Sissay (1998), Etana (2002) and Agyei, (2003) who indicated the highest
FEC during the rainy season and the lowest during the extreme dry season.
The lowest epg count obtained during the early wet season of this work could be explained by
the fact that, those larvae which get access to the host after first rain might have not been
reached to egg laying adults during the time of sampling, therefore their existence in high
number was not indicated by the presence of proportionally high number of eggs in the faeces
(Troncy, 1989; Jorgen and Brian, 1994).
46
The relatively low faecal egg count obtained in the lowland study site (Mirab-Abaya) during
the early dry period could be explained by the better opportunity goats have for browsing
during early dry season, therefore less challenge by the infective larvae and less nematode
burden during previous wet season (Smith, 1994).
The significantly higher (p<0.05) mean nematode egg count observed in sheep than goats in
late wet season (331.9±79.5 versus 261.8 ± 55.4) and the significantly higher epg out put
(p<0.05) of goats than sheep during early wet season (252.3 ± 46.6 in goats versus 192.6 ±
29.1 in sheep), might probably be due to the variations in feeding habit of the two species and
partly also due to the seasonal influence. Goats are browsers and they prefer browsing than
grazing in late wet season when the plant leaves are plenty, so that the challenge by the
infective stage of larva is less than that of sheep, which prefer to graze than browse at any
season. During the early wet season, however, due to the scarcity of plants for browsing and as
most of the plant leaves shaded out during the late dry season until few days after the rain; the
goats are equally exposed to the infective larvae present on newly emerged grass. Nonetheless,
as the immune status of goats is weaker than sheep (Craig, 1998; Radostits et al., 2000), the
establishment rate of infective larvae and the fecundity of adult female nematodes in goats
might be higher than those of sheep so that higher burden together with higher fecundity of
nematodes in goats revealed it self by higher nematode egg out put.
The significant variation (p<0.05) obtained in mean faecal egg out put of male animals
(293.4±43.4) than females (262.6±21.0) differs from the findings of previous works (Esayas,
1988; Achenef, 1997; Getachew, 1998) who indicated absence of difference in sex variation;
and from that of Assefa and Sissay (1998) who reported female animals to have higher faecal
egg out put than male animals. The difference of present finding might be related to the fact
that male animals in the study areas are sold with good market price as they have greater
demand for export to Arab countries and only few were allowed to remain in the flocks mainly
for the purpose of breeding service. The action and process of serving numerous breeding
females by few male animals in a flock, wastes their grazing time and energy and could lead to
stress unless they are supplemented, leading to increased establishment of infection and higher
nematode faecal egg out put in male animals of the study site.
47
The intensity of nematode infection, 58.4% light; 40.5% moderate and 1.1% heavy in
necropsied small ruminants of the present study differs from other findings in different regions
of the country (Melkamu, 1991; Dereje, 1992; Yoseph, 1993; Achenef, 1997; Haile-leul,
2002). The difference of present finding might be due to different categorization systems used.
The percent of poly parasitism from necropsied animals in this study (97.7%) agrees with the
work of Yoseph, (1993). The existence of more than one nematode species in a single host has
an additive pathogenic effect on the host and the pathogenecity is usually high when
Haemonchus, Trichostrongylus and Oesophagostomum are present together (Jorgen and Brian,
1994) and epidemiologically important as it creates a year round task for small ruminant
breeders.
A strong positive correlation (rs = 0.57, p<0.001) of mean total nematode burden to mean total
epg count both from slaughtered sheep and goats of the study area was observed during the
study period. A similar positive correlation was found in sheep of Debre Brehan (Tembley et
al., 1997) with a correlation coefficient of rs= 0.52. Silvestre et al. (2000) also found positive
correlation (rs=0.47; p= 0.02) in dairy goats and a higher positive correlation of r=0.74 was
found in young sheep (McKenna, 1981)
The correlation of mean epg to individual nematode species burden in this study was found to
be significant for three nematode species, T. colubriformis, (rs=0.50); H. contortus, (rs=0.45);
and Oe. columbianum, (rs=0.39) and p <0.001 for all correlations. This finding also agrees
with the findings of Tembley et al., (1997); Silvestre et al., (2000) and Craig, (1998) who
stated the positive correlation between faecal egg count and the worm burden of economically
important parasites such as Haemonchus contortus, T. colubriformis and Oe. columbianum.
48
6. CONCLUSION AND RECOMMENDATIONS
The study carried out in three different agro-ecological zones of southern Ethiopia during the
last nine months has revealed the wide spread existence of six nematode species (Oe.
columbianum, T. colubriformis, H. contortus, T. axei, T. ovis, B. trigonocephalum) and four
previously non-reported nematode species (Teladorsagia spp., T. probolurus, S. papillosus
and C. curticei) infecting sheep and goats in the area.
The over all high prevalence of GIT nematodiosis of small ruminants in all agro-ecology
suggests the importance of the problem in the study area in hampering the productivity and
health of small ruminants.
The assessment of seasonal dynamics in prevalence of total GIT nematodiosis in post mortem
has shown lack of significant changes over time, which indicates that in all the seasons of the
year, there was high prevalence in all agro-ecological zones of the study area. However, the
economically important nematode species such as H. contortus, T. colubriformis, T. axei and
Oe. columbianum have indicated significant differences in their prevalence in different
seasons of the year especially during the beginning of dry season and the beginning of wet
season. This seasonal difference was also shown in coprological examination.
Some nematode species such as H. contortus has significant variation on agro-ecological basis
(42.6% in highland versus 85.0% in midland) while Teladorsagia spp. and C. curticei were
identified only in highland agro-ecological zone. Others such as B. trigonocephalum and T.
ovis were of secondary importance in all agro-ecological zones of the study areas. Nematode
species such as T. probolurus, S. papillosus and C. curticei were existed with lower
prevalence. Most of the examined infections (97.7%) were of mixed type and of light and
moderate intensity (58.4% were light, 40.5% were moderate).
Mean nematode burden and mean nematode faecal egg count was varied seasonally; nematode
burden was significantly high during early dry and early wet seasons
From the results of the present study, GIT nematodiosis of sheep and goats in the study areas
was found to be one of the major problems that hampered efficient utilization of the available
small ruminant resources at hand and thus requires serious attention by all concerned bodies or
49
institution to properly address and give solutions to the problem. Therefore the following few
points are recommended based on the findings.
2. Supplementation of important nutrients (protein) especially during the dry season should
be exercised to avoid nutritional related stresses that affect host immunity and
susceptibility to parasitic infections.
3. Further studies covering all agro-ecological zones should be done in all regions of the
country, as there could be unidentified nematode species that could cause significant
economic losses.
50
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8. ANNEXES
No. Date Site(origin) Spp. sex Age BCS Qualitative epg Other
result parasites
observed
1
2
.
66
Annex 3. Chemicals and reagents used in the study
20% phenol crystals in mixture of 20% distilled water, 8ml. Glycerine, 16.5ml lactic acid
67
Annex 4. Prevalence (%) of GIT helminthes of sheep and goats in Ethiopia based on
coprological and post mortem examination.
3. Oromiya
68
Annex 5. Major nematode species and their prevalence in different regions of Ethiopia based on post mortem findings.
Region Haemonchus spp. Trichostrongyl Bunostomum Oesophagostomu Trichuris spp. Chaberti Source
us spp. spp. m spp. a ovina
Sheep Goat Sheep Goat Sheep Goat Sheep Goat Sheep Goat Sheep
Tigray 95.5 90.4 28.9 68.7 - 18.1 82.2 24.1 6.7 81.9 - Getachew (1998)
Amhara: Gondar 36.4 45.8 6.8 20.8 34.1 81.3 77.3 68.8 54.6 60.4 Gebreyesus (1986)
Kombolcha 83.8 63.4 40.7 79.6 63.4 Genene (1994)
D/Berhan 62.1 51.7 13.8 65.5 3.5 Achenef (1997)
Arsi&Wollo 66.8 89.4 34.4 57 83.2 4.0
Bekele etal.(1982)
Somale: Ogaden 93.6 83.8 36.1 16.6 32.0 59.4 53.0 61.1 34.1 Solomon (1987)
&Esayas (1988)
Oromia: E.Shoa 65.5 72.6 39.3 41.1 50.0 82.1 72.6 67.1 60.7 69.9 Melkamu (1991)
Asela 63.1 54.8 44.5 85.7 59.5 Yosef (1993)
Bale 59.2 44.1 38.2 19.1 55.3 85.3 76.3 67.7 61.8 60.3 Tesfalem (1985)
Wollega 88.2 29.4 35.0 53.0 76.5 Ahmed (1988)
Illubabor 58.0 43.9 26.0 22.0 44.0 34.0 86.0 63.0 24.0 26.0 Bayou (1992)
SNNPRS: 80.0 81.0 10.0 13.6 10.0 17.2 90.0 88.2 50.0 27.8 Dereje (1992)
Wolayta
61.6 54.8 22.1 21.4 41.9 35.7 74.4 66.8 36.0 28.6 Haileleul (2002)
E/Shoa, Harar, 98.8 49.6 (T. a), 42.4 (B. phle.) 92.0 (O. col.) 67.6 (T. ovis) Kasambara (1999)
88.4(T. colu.)
Afar, Sidamo.
East shoa 89.5 78.1(T. colu.) 77.1 (O. col) 54.0 (T. ovis) Dessalegn(1999)
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Annex 6. Determining the age of the goat (Mike Steel, 1996)
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Annex 8. Body condition scoring of the goat (Mike Steel, 1996)
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Annex 9. Body condition scoring of the sheep (Gatenby, 1991)
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Annex 10. Post-mortem examination procedures
1. As soon as possible after removing the alimentary tract from the body cavity, the
abomasal-duodenal junction should be ligatured to prevent transfer of parasites
from one site to the other.
2. Separate the abomasum, small intestine and large intestine.
3. Open the abomasum along the side of the greater curvature wash the contents in to
the bucket under the running water and make the total volume up to 4 litres, and
then the mucus membrane will be carefully rubbed with fingers to remove any
worms adhering to the mucus membrane.
4. After thorough mixing transfer duplicate 400ml. samples to suitably labelled
containers and preserve in 70% alcohol.
5. The small intestine is run off the mesentery, open it along its entire length and
wash the contents in the bucket. Treat as for the abomasal content.
6. The contents of large intestine are washed in to a bucket, passed through a coarse
mesh sieve (aperture of 2 – 3 mm) and any parasite present collected and
preserved in 70% alcohol.
Parasites collected and preserved are counted and identified based on the
morphological features described in MAFF, 1977; Soulsby, 1982; Jorgen, H. & Brian,
P. 1994; Urquhart, 1996 and Bowman, 1999.
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Annex 11. Worm counting procedure (Urquhart et al., 1996)
A. Gross characteristics
Abomasum
Small intestine
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Large intestine
B. Microscopic confirmation
Abomasum
Haemonchus
Male: Dorsal ray of bursa asymmetric; spicules barbed near the tip.
Female: Vulval flap, usually linguiform, present; gravid worm contains several
hundred eggs; ovary coiled around intestine.
Teladorsagia
Male: spicules slender, rod like (T. circumcincta) or stout with branch near middle (T.
trifurcata)
Trichostrongylus axei
Female: vuval flap absent; gravid worm contains 4 – 5 eggs pole to pole.
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Small intestine
Trichostrongylus
Male: spicules leaf shaped (T. vitrinus) or spicules with “stap” near tip
(T. colubriformis)
Strongyloides
Only females present; long oesophagus; ovary and uterus show twisted thread
appearance behind oesophagus; objectors absent.
Cooperia
Both sexes: small cephalic vesicles present, giving anterior end a cylindrical
appearance; prominent cuticular striations in oesophageal region.
Nematodirus
Male: Spicule long; slender and fused; with extended tip which is heart- shaped
(N.battus); lanceolate (N.filicollis); bluntly rounded (N.spathiger).
Bursa shows two sets of parallel rays (N. battus) or four sets (other species)
Female: large egg present; tip of tail is pointed (N. battus) or truncated with small
spine (other species).
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Large intestine
Trichuris
Tail of female is bow shaped and that of male is spirally coiled with one spicule.
Chabertia
Large bell shaped bucal capsule with out teeth and rudimentary leaf crowns.
Oesophagostomum
Relatively small bucal capsule; cephalic vesicle is with cervical groove behind. Leaf
crowns and cervical alae often present.
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Annex 13. Faecal Egg Counting Techniques
The method involves the use of a Mc Master egg counting chamber as described
below.
6. Pour off supernatant, agitate sediment and fill tube to previous level with floatation
solution.
7. Invert tube six times and remove fluid with pipette to fill both chambers of
McMaster slide.
8. Examine the two chambers and multiply the number of eggs by 50 to arrive at
number of eggs per gram of faeces (epg).
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Annex 14. Field pictures from sheep and goats grazing and browsing during different
seasons of the year.
Plates 1 and 2, goats browsing during late wet season of the year in lowland AEZ of
the study area during the study period.
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Plates 4 and 5. Sheep grazing in highland AEZ of the study site during late wet and
mid dry season of the study period.
Plate 6. Goats grazing in highland AEZ of the study site during late wet season of the
study period.
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Plates 7 and 8. Goats grazing during early dry and late dry seasons of the year in
midland AEZ of the study sites during the study period.
Plate 9. Numerous nodules on the large intestine of sheep slaughtered at a local hotel
of highland AEZ of the study site during the study period.
Arrows
pointing at nodules on large intestine due to arrested Oe. columbianum larva.
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Annex 15. Microscopic pictures of male bursa and spicules of C.curtiei, Teladorsagia
spp., T. axei and T. probolurus identified from the study sites during the study period.
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Plates 12 and 13. Teladorsagia spp. from sheep (High land).
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9. CURRICULUM VITAE
I. Personal Data
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V. Papers and publications
VII. Referrences
Dr Abebe Wossene
Associate Professor, AAU,FVM,
Dr Asseged Bogale
Associate Professor, AAU, FVM
Dr Terzu Daya
Laboratory head, Soddo regional Veterinary Laboratory, Southern Ethiopia,
Wolayta Soddo. POB.82, Tel.06-51-25-93
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10. SIGNED DECLARATION SHEET
I the under signed, declare that the thesis is my original work and has not been
presented for a degree in any university.
This thesis has been submitted for examination with my approval as University
Advisor.
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