Reading material for Basic Mechanisms of Diseases/S.

Wahyuni/Department of Parasitology/ Medical

Faculty, Hasanuddin University/2009

PROTOZOA
Structure, Classification, Growth, and Development
Introduction
The Protozoa are considered to be a subkingdom of the kingdom Protista, although in the
classical system they were placed in the kingdom Animalia. More than 50,000 species have been
described, most of which are free-living organisms; protozoa are found in almost every possible habitat.
The fossil record in the form of shells in sedimentary rocks shows that protozoa were present in
the Pre-cambrian era. Anton van Leeuwenhoek was the first person to see protozoa, using microscopes
he constructed with simple lenses. Between 1674 and 1716, he described, in addition to free-living
protozoa, several parasitic species from animals, and Giardia lamblia from his own stools.
Virtually all humans have protozoa living in or on their body at some time, and many persons are
infected with one or more species throughout their life. Some species are considered commensals, i.e.,
normally not harmful, whereas others are pathogens and usually produce disease.
Protozoan diseases range from very mild to life-threatening. Individuals whose defenses are able
to control but not eliminate a parasitic infection become carriers and constitute a source of infection for
others. In geographic areas of high prevalence, well-tolerated infections are often not treated to eradicate
the parasite because eradication would lower the individual's immunity to the parasite and result in a high
likelihood of reinfection.
Many protozoan infections that are inapparent or mild in normal individuals can be life-threatening
in immunosuppressed patients, particularly patients with acquired immune deficiency syndrome (AIDS).
Evidence suggests that many healthy persons harbor low numbers of Pneumocystis carinii in their lungs.
However, this parasite produces a frequently fatal pneumonia in immunosuppressed patients such as
those with AIDS. Toxoplasma gondii, a very common protozoan parasite, usually causes a rather mild
initial illness followed by a long-lasting latent infection. AIDS patients, however, can develop fatal
toxoplasmic encephalitis. Cryptosporidium was described in the 19th century, but widespread human
infection has only recently been recognized. Cryptosporidium is another protozoan that can produce
serious complications in patients with AIDS. Microsporidiosis in humans was reported in only a few
instances prior to the appearance of AIDS. It has now become a more common infection in AIDS patients.
As more thorough studies of patients with AIDS are made, it is likely that other rare or unusual protozoan
infections will be diagnosed.
Acanthamoeba species are free-living amebas that inhabit soil and water. Cyst stages can be airborne.
Serious eye-threatening corneal ulcers due to Acanthamoeba species are being reported in individuals
who use contact lenses. The parasites presumably are transmitted in contaminated lens-cleaning
solution. Amebas of the genus Naegleria, which inhabit bodies of fresh water, are responsible for almost
all cases of the usually fatal disease primary amebic meningoencephalitis. The amebas are thought to
enter the body from water that is splashed onto the upper nasal tract during swimming or diving. Human
infections of this type were predicted before they were recognized and reported, based on laboratory
studies of Acanthamoeba infections in cell cultures and in animals.
The lack of effective vaccines, the paucity of reliable drugs, and other problems, including difficulties of
vector control, prompted the World Health Organization to target six diseases for increased research and
training. Three of these were protozoan infectionsmalaria, trypanosomiasis, and leishmaniasis.
Although new information on these diseases has been gained, most of the problems with control persist.

Reading material for Basic Mechanisms of Diseases/S. Wahyuni/Department of Parasitology/ Medical

Faculty, Hasanuddin University/2009

Structure
Most parasitic protozoa in humans are less than 50 m in size. The smallest (mainly intracellular forms)
are 1 to 10 m long, but Balantidium coli may measure 150 m. Protozoa are unicellular eukaryotes. As
in all eukaryotes, the nucleus is enclosed in a membrane. In protozoa other than ciliates, the nucleus is
vesicular, with scattered chromatin giving a diffuse appearance to the nucleus, all nuclei in the individual
organism appear alike. One type of vesicular nucleus contains a more or less central body, called an
endosome or karyosome. The endosome lacks DNA in the parasitic amebas and trypanosomes. In the
phylum Apicomplexa, on the other hand, the vesicular nucleus has one or more nucleoli that contain
DNA. The ciliates have both a micronucleus and macronucleus, which appear quite homogeneous in
composition.
The organelles of protozoa have functions similar to the organs of higher animals. The plasma membrane
enclosing the cytoplasm also covers the projecting locomotory structures such as pseudopodia, cilia, and
flagella. The outer surface layer of some protozoa, termed a pellicle, is sufficiently rigid to maintain a
distinctive shape, as in the trypanosomes and Giardia. However, these organisms can readily twist and
bend when moving through their environment. In most protozoa the cytoplasm is differentiated into
ectoplasm (the outer, transparent layer) and endoplasm (the inner layer containing organelles); the
structure of the cytoplasm is most easily seen in species with projecting pseudopodia, such as the
amebas. Some protozoa have a cytosome or cell mouth for ingesting fluids or solid particles. Contractile
vacuoles for osmoregulation occur in some, such as Naegleria and Balantidium. Many protozoa have
subpellicular microtubules; in the Apicomplexa, which have no external organelles for locomotion, these
provide a means for slow movement. The trichomonads and trypanosomes have a distinctive undulating
membrane between the body wall and a flagellum. Many other structures occur in parasitic protozoa,
including the Golgi apparatus, mitochondria, lysosomes, food vacuoles, conoids in the Apicomplexa, and
other specialized structures. Electron microscopy is essential to visualize the details of protozoal
structure. From the point of view of functional and physiologic complexity, a protozoan is more like an
animal than like a single cell figure 1 shows the structure of the bloodstream form of a trypanosome, as
determined by electron microscopy.

Reading material for Basic Mechanisms of Diseases/S. Wahyuni/Department of Parasitology/ Medical

Faculty, Hasanuddin University/2009

Figure 1. Fine structure of a protozoan parasite, Typanosoma evansi, as revealed by transmission

electron microcopy of thin sections (Adapted from Vickerman K: Protozoology. Vol. 3 London School of
Hygiene and Tropical Medicine, London, 1977, with permission.)
Classification
In 1985 the Society of Protozoologists published a taxonomic scheme that distributed the Protozoa into
six phyla. Two of these phylathe Sarcomastigophora and the Apicomplexa--contain the most important
species causing human disease. This scheme is based on morphology as revealed by light, electron, and
scanning microscopy. Dientamoeba fragilis, for example, had been thought to be an ameba and placed in
the family Entamoebidae. However, internal structures seen by electron microscopy showed that it is
properly placed in the order Trichomonadida of flagellate protozoa. In some instances, organisms that
appear identical under the microscope have been assigned different species names on the basis of such
criteria as geographic distribution and clinical manifestations; a good example is the genus Leishmania,
for which subspecies names are often used. Biochemical methods have been employed on strains and
species to determine isoenzyme patterns or to identify relevant nucleotide sequences in RNA, DNA, or
both. Extensive studies have been made on the kinetoplast, a unique mitochondrion found in the
hemoflagellates and other members of the order Kinetoplastida. The DNA associated with this organelle
is of great interest. Cloning is widely used in taxonomic studies, for example to study differences in
virulence or disease manifestations in isolates of a single species obtained from different hosts or
geographic regions. Antibodies (particularly monoclonal antibodies) to known species or to specific
antigens from a species are being employed to identify unknown isolates. Eventually, molecular

Reading material for Basic Mechanisms of Diseases/S. Wahyuni/Department of Parasitology/ Medical

Faculty, Hasanuddin University/2009

taxonomy may prove to be a more reliable basis than morphology for protozoan taxonomy, but the
microscope is still the most practical tool for identifying a protozoan parasite. Table 1 lists the medically
important protozoa.

Table 1. Classification of parasitic protozoa and associated disease

Life Cycle Stages
During its life cycle, a protozoan generally passes through several stages that differ in structure and
activity. Trophozoite (Greek for animal that feeds) is a general term for the active, feeding, multiplying
stage of most protozoa. In parasitic species this is the stage usually associated with pathogenesis. In the
hemoflagellates the terms amastigote, promastigote, epimastigote, and trypomastigote designate
trophozoite stages that differ in the absence or presence of a flagellum and in the position of the
kinetoplast associated with the flagellum.
A variety of terms are employed for stages in the Apicomplexa, such as tachyzoite and bradyzoite
for Toxoplasma gondii. Other stages in the complex asexual and sexual life cycles seen in this phylum
are the merozoite (the form resulting from fission of a multinucleate schizont) and sexual stages such as
gametocytes and gametes. Some protozoa form cysts that contain one or more infective forms.
Multiplication occurs in the cysts of some species so that excystation releases more than one organism.

Reading material for Basic Mechanisms of Diseases/S. Wahyuni/Department of Parasitology/ Medical

Faculty, Hasanuddin University/2009

For example, when the trophozoite of Entamoeba histolytica first forms a cyst, it has a single nucleus. As
the cyst matures nuclear division produces four nuclei and during excystation four uninucleate metacystic
amebas appear. Similarly, a freshly encysted Giardia lamblia has the same number of internal structures
(organelles) as the trophozoite. However, as the cyst matures the organelles double and two trophozoites
are formed. Cysts passed in stools have a protective wall, enabling the parasite to survive in the outside
environment for a period ranging from days to a year, depending on the species and environmental
conditions. Cysts formed in tissues do not usually have a heavy protective wall and rely upon carnivorism
for transmission. Oocysts are stages resulting from sexual reproduction in the Apicomplexa. Some
apicomplexan oocysts are passed in the feces of the host, but the oocysts of Plasmodium, the agent of
malaria, develop in the body cavity of the mosquito vector.
Reproduction
Reproduction in the Protozoa may be asexual, as in the amebas and flagellates that infect
humans, or both asexual and sexual, as in the Apicomplexa of medical importance. The most common
type of asexual multiplication is binary fission, in which the organelles are duplicated and the protozoan
then divides into two complete organisms. Division is longitudinal in the flagellates and transverse in the
ciliates; amebas have no apparent anterior-posterior axis. Endodyogeny is a form of asexual division
seen in Toxoplasma and some related organisms. Two daughter cells form within the parent cell, which
then ruptures, releasing the smaller progeny which grow to full size before repeating the process. In
schizogony, a common form of asexual division in the Apicomplexa, the nucleus divides a number of
times, and then the cytoplasm divides into smaller uninucleate merozoites. In Plasmodium, Toxoplasma,
and other apicomplexans, the sexual cycle involves the production of gametes (gamogony), fertilization to
form the zygote, encystation of the zygote to form an oocyst, and the formation of infective sporozoites
(sporogony) within the oocyst.
Some protozoa have complex life cycles requiring two different host species; others require only
a single host to complete the life cycle. A single infective protozoan entering a susceptible host has the
potential to produce an immense population. However, reproduction is limited by events such as death of
the host or by the host's defense mechanisms, which may either eliminate the parasite or balance
parasite reproduction to yield a chronic infection. For example, malaria can result when only a few
sporozoites of Plasmodium falciparumperhaps ten or fewer in rare instancesare introduced by a
feeding Anopheles mosquito into a person with no immunity. Repeated cycles of schizogony in the
bloodstream can result in the infection of 10 percent or more of the erythrocytesabout 400 million
parasites per milliliter of blood.
Nutrition
The nutrition of all protozoa is holozoic; that is, they require organic materials, which may be particulate or
in solution. Amebas engulf particulate food or droplets through a sort of temporary mouth, perform
digestion and absorption in a food vacuole, and eject the waste substances. Many protozoa have a
permanent mouth, the cytosome or micropore, through which ingested food passes to become enclosed
in food vacuoles. Pinocytosis is a method of ingesting nutrient materials whereby fluid is drawn through
small, temporary openings in the body wall. The ingested material becomes enclosed within a membrane
to form a food vacuole.
Protozoa have metabolic pathways similar to those of higher animals and require the same types of
organic and inorganic compounds. In recent years, significant advances have been made in devising
chemically defined media for the in vitro cultivation of parasitic protozoa. The resulting organisms are free

Reading material for Basic Mechanisms of Diseases/S. Wahyuni/Department of Parasitology/ Medical

Faculty, Hasanuddin University/2009

of various substances that are present in organisms grown in complex media or isolated from a host and
which can interfere with immunologic or biochemical studies. Research on the metabolism of parasites is
of immediate interest because pathways that are essential for the parasite but not the host are potential
targets for antiprotozoal compounds that would block that pathway but be safe for humans. Many
antiprotozoal drugs were used empirically long before their mechanism of action was known. The sulfa
drugs, which block folate synthesis in malaria parasites, are one example.
The rapid multiplication rate of many parasites increases the chances for mutation; hence,
changes in virulence, drug susceptibility, and other characteristics may take place. Chloroquine
resistance in Plasmodium falciparum and arsenic resistance in Trypanosoma rhodesiense are two
examples.
Competition for nutrients is not usually an important factor in pathogenesis because the amounts utilized
by parasitic protozoa are relatively small. Some parasites that inhabit the small intestine can significantly
interfere with digestion and absorption and affect the nutritional status of the host; Giardia and
Cryptosporidium are examples. The destruction of the host's cells and tissues as a result of the parasites'
metabolic activities increases the host's nutritional needs. This may be a major factor in the outcome of
an infection in a malnourished individual. Finally, extracellular or intracellular parasites that destroy cells
while feeding can lead to organ dysfunction and serious or life-threatening consequences.
Resume
Protozoa are one-celled animals found worldwide in most habitats. Most species are free living,
but all higher animals are infected with one or more species of protozoa. Infections range from
asymptomatic to life threatening, depending on the species and strain of the parasite and the resistance
of the host.
Protozoa are microscopic unicellular eukaryotes that have a relatively complex internal structure
and carry out complex metabolic activities. Some protozoa have structures for propulsion or other types
of movement.
On the basis of light and electron microscopic morphology, the protozoa are currently classified
into six phyla. Most species causing human disease are members of the phyla Sacromastigophora and
Apicomplexa.
The stages of parasitic protozoa that actively feed and multiply are frequently called trophozoites;
in some protozoa, other terms are used for these stages. Cysts are stages with a protective membrane or
thickened wall. Protozoan cysts that must survive outside the host usually have more resistant walls than
cysts that form in tissues.
Binary fission, the most common form of reproduction, is asexual; multiple asexual division
occurs in some forms. Both sexual and asexual reproduction occur in the Apicomplexa.
All parasitic protozoa require preformed organic substancesthat is, nutrition is holozoic as in
higher animals.
References
1. Englund PT, Sher A (eds): The Biology of Parasitism. A Molecular and Immunological Approach.
Alan R. Liss, New York, 1988 .
2. Goldsmith R, Heyneman D (eds): Tropical Medicine and Parasitology. Appleton and Lange, East
Norwalk, CT, 1989 .

Reading material for Basic Mechanisms of Diseases/S. Wahyuni/Department of Parasitology/ Medical

Faculty, Hasanuddin University/2009

3. Lee JJ, Hutner SH, Bovee EC (eds): An Illustrated Guide to the Protozoa. Society of
Protozoologists, Lawrence, KS, 1985 .
4. Kotler DP, Orenstein JM. Prevalence of Intestinal Microsporidiosis in HIV-infected individuals
referred for gastrointestinal evaluation. J Gastroenterol. 1994; 89: 1998.
5. Neva FA, Brown H: Basic Clinical Parasitology, 6th edition, Appleton & Lange, Norwalk, CT, 1994
.