PROTOZOOLOGY

Protozoology is that branch of biology science, which deals with unicellular animal
life known as protozoa. The history of protozoa dates back to 1674, when Dutch
naturalist, Leeuwenhoek described them as animalcules. The term Protozoa was applied
by Goldfuss in 1920.

Morphology of the protozoa: All protozoa consist of a eukaryotic type of cell. Body of
the protozoa consists two parts i.e., nucleus and cytoplasm.

Nucleus: Most protozoans have a single nucleus, which is known as vesicular nucleus.
But in some, such as B. coli there are two nuclei. In this case one is small in size –
known as micronucleusor vesicular nucleus. This is similar to vesicular nucleus of the
uninucleated protozoa and it performs the function of reproduction. The second large
in size nucleus is known as meganucleus or macronucleus or massive nucleus. This is
usually kidney shaped and it performs the metabolic function of protozoa. Opalina is a
multinucleated organism and contains about two to hundred similar sized nuclei. Other
different characters of vesicular nucleus are a clear nuclear membrane is present and
at the center and there is a nucleolus generally called karyosome. In case of mega or
massive nucleularge number of chromatic granules are scattered through out the
nucleoplasm.

Cytoplasm: It is extranuclear part of

the protozoa. This is again divided
into two parts outer homogenous
ectoplasm and inner granular
endoplasm. The endoplasm contains
food vacuoles and pigmented
granules. In non parasitic protozoa
there is specific organelles known as
contractile vacuole which is
responsible for excretion of waste
materials. But in most of the
parasitic protozoa it is absence.
The ectoplasm is homogenous and
sometimes protected by thin cuticlar
structure known as pellicle.

Locomotion of protozoa: The protozoa may move either by gliding or by means of

locomotory organs. Gliding movement is seen in case of Toxoplasma and Sarcocystis. They
do not have any locomotory organelles. On the body surface there are numerous
ridges like structures which are known as microtubules which help in gliding movement.
But other protozoan can move with the help of different types of locomotory organs
such as pseudopodia, flagella and cilia.

Pseudopodia: The pseudopodia are finger like projections (Temporary finger like
projections). These are produced only at the time of requirement. These types of organelles
are found in Amoeba.
Flagella: These are filamentous structure, very fine and highly mobile in nature. Each
flagellum consists of a central axial rod known as axoneme, which is surrounded by a
cytoplasmic sheath. The axoneme arises from a structure known as kinetoplast, which
consists of a nucleus or chromatin mass known as kinetonucleus or parabasal body. Near this
kinetoplast another granular substance is present known as blepheroplast or basal granule. In
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some protozoa the flagellum runs the side of the body and is attached to the body by means of
delicate membrane known as undulating membrane.
Cilia: These are also very fine short and small hairs like structure. They arise from the
ectoplasm. They are more in number and coverers entire body of the ciliate protozoa. A part
from movement they also help in the ingestion of food materials and other function such as in
reproduction.

Nutrition of protozoa: In case of Amoeba the food materials are engulfed by means of
pseudopodia. In case of B. coli there is special opening known as cytostome which acts as the
mouth and food material are taken through this cytostome. In case of Apicomplexa or
Sporozoa protozoans there is a special opening towards the side of anterior part of the body
known as micropore or micropyle through which the food materials are taken. In other
protozoan food materials are absorbed through the body surface. In all protozoa these food
materials enter into a vacuole in which digestion takes place. In B. coli and E. histolytica the
used host cells as a food material and in case of E. coli the food material is absorbed through
the host intestine.

Types of nutrition: Broadly two main types of nutrition are seen:

1. Holozoic nutrition: In case of parasitic and commensal protozoa ready made or
preformed food material derived from the body of the host.
2. Holophytic nutrition: In this case food is synthesized with the help of chlorophylls
carried on the chromatophore in sunlight. The holozoic nutrition is again divided into
number of types.
a. Saprozoic nutrition: This is absorbed nutrient through the body wall, these being
utilized directly by the organisms.
b. Coprozoic nutrition: In this case the protozoan takes organic food from the faeces. c.
Parasitic nutrition: - This type of nutrition only found in protozoan parasites.

Respiration: Most of the protozoa absorb oxygen through the body surface and remove
carbondioxide through the respiratory vacuole.

Excretion: The waste materials are excreted through the contractile vacuole in case of
freeliving or none parasitic protozoa. In case of parasitic protozoa such as B.coli a spiral
opening is present which is known as cytopyge through which waste materials are excreted.
In other parasitic protozoa the waste materials are excreted as small vacuoles containing
fluids. These vacuoles come out through the body surface. Parasites not only eat but also
drink. It takes fluids into small vacuoles even when it is not feeding. This method of
ingestion is known as pinocytosis.

Reproduction: Reproduction in parasitic protozoan usually occurs by two methods

1. Asexual and 2. Sexual.

1. Asexual reproduction is divided into number of types.

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 a. Binary fission: Binary fission is the form of asexual reproduction. In this two
daughter cells result from a parent cell. In most of protozoans division commonest takes
place along the longitudinal axis known as longitudinal binary fission. But in ciliates like
B. coli division is along the transverse axis. In this case the nucleus divides first then the
cytoplasmic division takes place.

b. Multiple fission or schizogony: In this case from one individual more than two
individuals are produced. This occurs by the mitotic division of nucleus into a number
of small fragments. Each fragment is surrounded by a portion of the cytoplasm. The
division form is known as a schizont and the daughter form is known as merozoites. The
process of the division of nucleus upto the formation of merozoites is known as
schizogony.

c. Budding: It is an asexual reproductive process in which two or many daughter forms

are produced by the parent cell.
There is usually an unequal fragmentation of the nucleus and cytoplasm, but the budded
forms are separated off and then developed to full size.

d. Endopolyogeny: It is a form of asexual multiplication or internal budding; whereby new

progeny are formed within the parent cell. E.g. Toxoplasma, Sarcocystis etc.

e. Endodyogeny: It is a simple form of endopolyogeny in which division occurs inside the

mother individual i.e. two daughter individual are formed inside the mother cell later they are
separated from the mother cell. It is seen in forms such as Toxoplasma and Sarcocystis.

f. Sporogony: This is a process of multiple fission after sexual reproduction or syngamy. This
division normally follows syngamy and a number of sporozoites are formed within the walls
of a cyst.

2. Sexual division:
a. Conjugation: It is a form of sexual reproduction, which occurs in the ciliates. In this,
two organisms are paired and exchange their nuclear material. The individuals separate
and nuclear reorganization takes place. e.g., B. coli

b. Syngamy: It is a sexual reproduction in which two gametes fuse to form a zygote.

The male gamete is a microgamete and the female is called macrogamete, which are
produced from microgametocytes or microgamonts and macrogametocytes or
macrogamonts,
respectively. The process of gamete formation is gametogony and the gametes may be similar
in size i.e., called isogamy or may markedly differ called anisogamy.

Somecommon term used in protozology:

1. Trophozoite: This is the vegetative form of
protozoa, which feeds, grows and ultimately
divides to form daughter individuals, which
repeat the same process.

2. Cyst: It is the immotile form of protozoa

which are protected by a cyst wall. The
encystation may occur before or after fertilization
and they may be transmitted from one individual
to other without multiplication e.g., cyst of E.
histolitica, Giardiaand oocyst of Coccidia.

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3. Spores: These are the multinucleated forms, developed inside the encysted zygote
during a process known as sporogony and are called spores which produce sporozoites
e.g., Coccidia

PHYLUM: SARCOMASTIGOPHORA
SUBPHYLUM: SARCODINA
Family 1: Endamoebidae
Genus: Entamoeba
Species: E.histolytica and E.coliLocation and host: It is found in the caecum and colon of man,
pig, dog, cat, monkey and sometimes rat. The disease caused by this protozoon is called Amoebic
dysentery or Amoebiasis.
Morphology: It is found in two forms; first is
trophozoite or vegetative and second is cystic form.
Structure of trophozoite: These are active forms and
are found as two varieties – large and small. Large
forms are more pathogenic and size is more than 20-30
µm in diameter. They have crawling type of movement
with the help of pseudopodia. These vegetative forms
sometime passed out in the dysenteric stool. The
cytoplasm has a clear ectoplasm and endoplasm. The
endoplasm has foodvacuole, which contains host cells,
RBC and sometime bacteria. The nucleus is single,
spherical with an endosome or nucleolus present in the
center. The nucleus membrane is lined by a row of fine
chromatin granules.
Morphology of cyst form-The cysts are spherical or subspherical in shape and size is 5-20µm in
diameter. These cysts have a double layer cyst wall and these cysts are passed out in
constipated stool. The immature stages contain 1-2 nuclei, glycogen granules and chromatoid rods.
This glycogen granule and chromatoid rods act as reserve food material. The mature cyst has
four nuclei and diffused glycogen granules but no chromatoid rods. In some cases, they disappear
when they are consumed.

Life cycle: The cysts are passed out in faeces or when the uninucleate cyst form is passed out,
they grow into mature cysts. When these cysts are ingested by the host with contaminated food
and water then the host is infected. In the intestine some more nuclear division and
cytoplasmic divisions take place and four nuclei form; then eight nucleated forms are formed.

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On rupture of cyst wall, these individuals having uninucleate bodies come out and these are known
as amoebulae. They further pass into the large intestine and develop into trophozoites.

Multiplication: The larger trophozoites (more pathogenic), multiply by binary fission and
penetrate into the wall of large intestine by means of pseudopodia and there cyst formation
also takes place. Before turning into a cyst, trophozoites change into smaller forms and expel
out all the food vacuoles. They become round and stop feeding. Immature stages have 1-2
nuclei and mature cyst have four nuclei after division.

Pathogenicity and symptoms: Large varieties of E. histolytica are more pathogenic. After
being excysted in the intestine they penetrate into the intestinal mucosa of large intestine by
dissolving the surface (mucosa, submucosa) epithelium with the help of proteolytic enzyme,
trypsin and pepsin. They penetrate into the intestinal mucosa to form colonies which
penetrate further deep into the submucosa and spread. Due to this fast spreading, flask shaped
ulcers are produced with narrow ends directed towards the lumen. Sometimes they penetrate
deep into muscularis mucosa of large intestine and reach lymphatics and blood vessels. Then
they are carried to different soft organs of the body inside which they form ulcers. Right lobe
of the liver can be highly affected. Other organ such as lungs and brain may also be affected.
Symptoms: From the pathogenesis it is known that there are two types of disease duet to this
parasite - Intestinal forms and Extra-intestinal forms.
Intestinal form: It may be of two types – Acute and Chronic

Acute: Symptoms: - Dysentery. Abdominal pain. Nausea and vomiting. Loss of appetite.
Stool may be mixed with blood and mucus. There is rectal pain and
some time fever is seen.
Chronic form: Any type of symptoms may be seen such as Abdominal pain.
Intestinal ulcer. Headache.
Nausea and vomiting. There is bowel
irregularity.
Extraintestinal form: Abscesses in liver may produce pain in the liver; lesion may be more in
case of right side of liver. There is high fever and the leucocytes count increases, some
timebrain and lung abscess are also found.

Diagnosis: It is done by:

1. Examination of faecal sample and finding the cyst or trophozoites. For this purpose 1-2%
iodine solution; containing solution of iodine and 1% KI is put into the smear and then
examined under the microscope. Iodine solution stains the nucleus.
2. From clinical symptoms.
3. In chronic case faecal culture may be done in which Amoeba develop in
the culture.
4. Liver abscess detected from X-ray.
5. Blood sample examination reveals high leukocyte count.
6. There are serological methods such as complement fixation test.

Treatment:
For intestinal form:
a. The drug of choice is Metronidazole-400mg every eight hour for 5days in
chronic cases in human and 800mg every eight hours for 5days in acute cases in
human; 20mg /kg /day for 10days in animals
b. Niridazole - 25mg/kg daily for 5-10 days in adult human.
For extraintestinal form:
a. Emetine hydrochloride- 65mg dissolved in 1ml of distilled water and provided
deep I/m or s/c injection for 5-10 days in human and dog.

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 b. Chloroquine - 300mg once daily orally for 5days in human and animal.

Control: 1. Good sanitation.

2. Improved sewage disposal.
3. Avoid contamination of faecal material in the food material.
4. Improvement of personal hygiene.
Amoebiasis is a primary disease of man. Although the parasites are found in domesticated
animals, they get this infection from human faeces and clinical type of infection is rarely seen
in domesticated animal.

E. coli
It is non-pathogenic amoeba found in the caecum and colon of man and other animals. It is
world wide in distribution being commoner in warm, moist climates. The morphological
characters to differentiate it from E. histolytica are:
Characters E. histolytica E. coli
Vegetative form or Trophozoite form
Size 20-30µm 20-40µm
Motality Active & progressive Slugish & no progressive
movement

Inclusion in the Only RBC in fresh

Bacteria
specimen
are present
food vacuole

Nucleolus Situated at the centre of Excentrical

nucleus (Eccentric)

Nuclear membrane Inner surface is lined Lined by

by row of fine coarse
chromatin granules chromatin granules.

Size 5-20 µm 15-30µm

Shape Spherical Spherical
Cytoplasm Greenish yellow Yellow brown
Chromatin rod Thick rod like with
Filamentous
rounded with
ends pointed
or square shape.

Nuclei 4 nuclei in mature cyst 8 nuclei in mature cyst

Nucleolus Centrally present Excentriacally present

E. gingivalis
These are found in the gums of man, dog, cat, and monkey etc. They sometimes cause
pyorrhoea. In this case no cyst formation takes place. Only the trophozoite form is seen
which is 5-35µm in diameter (usually 10-20µm in diameter). They produce multiple
pseudopodia, which are blunt and large. The chromatin granules lining the inner surface of
nucleare membrane are small and continuous. They are transmitted by fomite infection.

E. bovis
This parasite is found in cattle. The sizes of trophozoites are 5-20µm. Its cytoplasm is filled
with vacuoles. Nucleus is large and with a large central endosome made up of a compact
mass of granules. The cyst is uninucleated and size is about 4-15um in diameter.
Amoebae with 8 nucleated cyst - E. coli
" " 4 " " - E. histolytica, E. hartmanni, E. equi etc.
" " 1 " - E. bovis, E. ovis, E. suis etc.
" " unknown " " " - E. gingivalis, E. canibuccalis etc.
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 SUBPHYLUM: MASTIGOPHORA
CLASS: ZOOMASTIGOPHOREA
ORDER: KINETOPLASTIDEA
Family 2-Trypanosomatidae
These are leaf like or rounded organisms. Only two genera are important in veterinary field.
They are Trypanosoma and Leishmania.

General characters of the family Trypanosomatidae:

These are commonly known as Trypanosome and they are found in blood and tissue fluid but
rarely on the tissue of the mammals and birds. During their life cycle most of the parasites
show at least one further developmental stage which is undergone in the insect vector and
these stages are morphologically different from each other. These stages are:
Amastigote Promastigote Epimastigote Trypomastigote Opisthomastigote Choanomastigote
Previously Leishmanial form Leptomonad form
Crithidia form or barley corn form Trypanosome form
Trypanosome or trypanomorphic form or Herpetomonad form Barley corn form

The different characters of these stages are

Characters Amastigote Promastigote Epimastigote Trypomastigote

Shape Circular or oval Leaf like or Leaf like or Leaf like or

elongated elongated elongated

Nucleus Central or At the middle of the At th posterior At the middle of

position peripheral body end of the body the body

Kinetoplast Rod shaped or Dot like and Dot like but Dot like
present in front of present anteriorly. present at the middle present posterior
the nucleus Towards the of the body at to the nucleus.
nucleus at the the anterior side of the
anterior tip of the body. nucleus
Flagellum and Absent Flagellum present but Flagellum and small Flagellum and
undulating membrane undulating undulating undulating
membrane absent membrane present membrane well
developed
Host Vertebrates Invertebrate such Arthropods Vertebrates
as arthropods

Development Develop into Develop into Develop into May develop into
promastigote form amastigote form inside promastigote form inside previous 3 stages.
inside the insect the vertebrate host. the arthropod vector.
vector

Genus Trypanosoma
These are found in the blood and tissue
fluids and a few species are also found
inside the tissue of mammals, birds and
reptiles. These parasites may be
polymorphic or monomorphic.

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 Polymorphic - The same species has
numerous forms such as elongated,
epimastic and metacyclic form in insect
host.

Monomorphic - It does not change the shape

and size throughout their development.
These are transmitted by insect vectors.

Both forms are usually transmitted by insect vectors. With vectors mainly two things may
happen. Any Trypanosome can be mechanically transmitted, that is, there may be no cyclical
development in arthropods. Or, according to cyclical development, Trypanosoma has two
types of development cycles (classification according to development cycle) inside the insect
vector.
a. Anterior station development
b. Posterior station development

a. Anterior Station Development or Salivaria- The kinetoplast is small and more

terminal or subterminal, posterior extremity is blunt, may be flagellum free, undulating
membrane is well developed.

i) Brucei group- All are polymorphic; intracellular forms have been found. In some species
parasite is long, intermediate and stumpy in form and they are found in the intracellular tissue
fluids and cyclical development is present in mid gut, proboscis and salivary gland of
Glossina spp.

T. brucei- This parasite develops on the posterior part of the mid gut of G. morsitan. It is a
parasite of sheep, goat and other domesticated animals and in antelops. The parasite causes
Nagana disease in domesticated animals in West Coast of Africa.

T. gambiense- This parasite shows polymorphic forms. It is found in man and also in
antelopes. It causes African Sleeping Sickness in humans in West Coast of Africa.

T. rhodesiense- This parasite causes Encephalitis, Keratitis in cattle, sheep, goat etc, and
Trypanosomiasis in man in South Africa. Both strains are transmitted from wild antelops or
domesticated animals to man.

ii) Congolense group- Development of parasite is in mid gut and proboscis of G. palpalis;
free flagellum is absent. Parasites are intracellular forms, they are monomorphic or
polymorphic.

T. congolense - Found in cattle, sheep, horse and pig. It is monomorphic.

T. dimorphic - Found in cattle, sheep, horse and pig. It is monomorphic.

T. simiae - Most pathogenic in pig. Camel, horse and cattle are also infected. They are
polymorphic.

iii) Vivax group- Monomorphic development occurs only in the proboscis of Tse-tse fly and
free flagellum is present and high sugar utilized.
T. vivax- Cattle, sheep, goat and antelope.
T. uniformae- Cattle, sheep, goat and antelope.

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iv) Evansi group - Although it belongs to salivaria no cyclical development takes place inside
the insect host and they are transmitted mechanically. All species show polymorphism.
T. evansi- Only found in the proboscis of Stomoxys, Tabanus etc. and survive only 10-15
minutes. They cause Surra disease and infect cattle, horse, dog, camel and elephant.

T. equinum - They cause Mal de caderas disease or weakness of the hinder quarters result in
a staggering gait in horses.

T. equiperdum - Causes venereal diseases (Stallion's disease or Dourine) in horse and mule.

b. Posterior Station Development or Stercoraria- Parasite consists of a large kinetoplast

which is not in terminal in position. The posterior extremity is tapering, free flagellum is
present, undulating membrane is not well developed. This section consists only one group.

1) Lewisi group
T. lewisi - Rat acts as the host, epimastigote and mastigote stages are found.
T. theileri - Cattle acts as the host, in ordinary condition it is non-pathogenic.
T. melophagium - Transmitted by sheep ked in sheep not so pathogenic.
T. rangeli - Dogs acts as host.
T. gallinarum - Hosts are fowl and other birds found in South East Asia.
T. avium - Hosts are fowl and other birds found in South East Asia, Europe, Canada, and
N. America. All are nonpathogenic except T. cruzi.
T. cruzi - Causes "Chagas" disease in man and dog in South America.

Out of these groups only the Evansi group are the pathogenic, which is found in the Indian
subcontinent and Far East, North East and in North Africa, Phillippines and Central and
South America. Other species are transmitted by Glossina spp., which is not found in Nepal
and India. Most of the species are naturally transmitted by the cyclical development inside
the insect vector. A few spp. of Trypanosome can be transmitted mechanically.

Developmental stages
Inside the definitive host- After the metacyclic form of parasites are injected into the blood of
host by infected arthropods vector, they change into trypomastigote form in blood and
multiplication occur by longitudinal binary fission. No sexual process has been observed.
Division commences at the kinetoplast followed by the nucleus and then cytoplasm. The
flagellum donot divide and remains attached to one individual and second daughter individual
develops new flagellum. Later trypomastigote form changes into intermediate short stumpy
form, which is carried by the insect host.

Development inside the insect vector. The Evansi spp. of Trypanosome does not undergo
any cyclical development inside the insect vector and they are transmitted mechanically. For
this mechanical transmission the feeding habit of fly should be such that it bites another host
immediately after sucking blood from the infected host. In the cyclical development the
development may be anterior station or posterior station. The anterior station development
occurs inside the mid gut, proventriculus, salivary gland and proboscis of insect. Posterior
station development occurs in the hindgut of the insect.
Anterior Station Development- Three groups of Trypanosome develops in the anterior
station of the flies. Out of these the Brucei group develops in the midgut, proventriculus and
salivary gland. The Congolense group develops in midgut and proboscis and Vivax group
develops only in the proboscis. In this case (Anterior Station Development) Glossina flies
acts as intermediate host. In the polymorphic trypanosome, the stumpy forms of
trypanosomes are responsible for initiating development in the insect vector. There they
change into elongated forms of trypomastigote form and then to epimastigote form inside the
insect vector and then in the proboscis of insect vector they develop into metacyclic form of

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trypomastigote form. It is the infective form and it resembles to stumpy form of
tryponastigote found in definite hosts e.g. T. brucei, T. congolense and T. vivax developed in
Glossina fly.
Posteriror Station Development- Only one group – Lewisi develops in this manner.
Intermediate or insect vector may be different for different species such as fleas, bugs -bed
bug (Reduviid bug) and Triatoma spp. of bug (Kissing bug), Triatoma spp. acts as vector of
T. cruzi, Melophagus ovinus (sheep ked). In this case development occurs in hindgut and they
develop into epimastigote form and then into metacyclic trypanosome. These metacyclic
trypanosomes are paddled out in the faeces, infection occurs after contaminating the skin
wounds by the faeces. E.g. are T. cruzi, T. lewisi etc. that develop in the body of Kissing bug
and flea.

Pathogenesis and symptoms-

1. The bites of fly or insect causes small papillae formation on the skin with skin
irritation.
2. The multiplication of Trypanosome in the blood of host produces the following
symptoms-
 Pyrexia (rise of temperature) Progressive anaemia
 Paleness of visible mucous membrane e.g. lip and conjunctiva
 Oedema of the dependent parts of the body
 There is emaciation and animal is unable to rise.
 This makes it difficult for the animal which leads to death.
 There is decrease in potassium content of blood and increase in
sodium.
 There is a decrease in the leukocytic count, which may cause .
 The pH of the blood decreases and it becomes up to 5.5.
 There is decrease in albumin content of blood and an increase in euglobulin contents.

Changes in the nervous system

In some case the Trypanosome penetrates through the blood capillary and reaches the
cerebro-spinal fluid and cause Encephalomeningitis. In the brain parenchyma they form
Hyaline vacuolated cells known as mononuclear cells of central nervous system.

Causes of death in case of trypanosomiasis-

The actual cause of death is not known but death may be due to following reasons:
1. Hypoglycemia-The Trypanosome consumes lot of blood sugar due to which there
is hypoglycemia (less sugar).

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 2. Asphyxia or hypoxia-This is due to decrease in blood sugar level due to which
there is increase in lactic acid production and there is lesser absorption of oxygen
by Hb due to which there is difficult in respiration.

3. Stress factor- Loss of glucose shows stress on the liver resulting in dysfunction of liver and
toxemia.

4. Endotoxins- Disintegration of Trypanosome may result in the formation of

endotoxins, this endotoxin is proteolytic in nature and death may occur due to this
endotoxin.

Postmortem changes-
 Enlargement of spleen, liver, lymph gland
 Bone marrow is congested
 There is gelatinous infiltration of the subcutaneous tissue
 Pinpoint haemorrhages of visible mucous membrane and sub cutaneous tissue
 There is congestion and haemorrhage in stomach and intestine
 In the eye there is keratitis and conjunctivitis.

Diagnosis
1. Microscopic- Examination of blood, serous fluid and cerebro-spinal fluid, often
stained with Giemsa stain and Leishmania stain.
2. Biological - 1-5ml of affected blood may be injected intra-peritoneally or I/V into
the susceptible laboratory animal. Such as rat, mice, guinea pigs. Rat and guinea pig
shows the symptoms within 2-3 days. Guinea pigs and rabbits take 1-2 weeks to
show the symptoms.
3. Cultural- Non pathogenic Trypanosomes can be cultured in NNN medium (Novy
and MacNeal Nicolle). All Trypanosomes can be cultured in chick embryo.
4. Serological-
i. Formal gel test- When a drop of formalin is added to 1ml of suspected
serum, in positive case gel formation takes place.
ii. Mercuric Chloride test- It is specific for camel when drop of 1:25000 or
1:30000 dilution of mercuric chloride is added to 1 drop of suspected
serum while precipitation takes place within few minutes indicating the
positive reaction.
iii. Stilbamide test- It is specific for cattle serum from suspected animal is
collected and kept for 48 hrs. In refrigerator when a drop of this serum is
added to 1 ml of 0.3% solution of stilbamide in distilled water, the following
reaction takes place. In strongly positive cases the serum coagulates
immediately (after one or two minutes) and sinks to the bottom as a large
mass and it again dissolves within 5 minutes. In mild positive test the serum
drops sinks to the bottom but it dissolves slowly. In light positive case the
serum drops sinks to the bottom as a fine thread and it dissolves slowly. If
negative the serum drop is coagulated on the surface and dissolves quickly.

5. Xenodiagnosis- The insect vector is allowed to bite the suspected animal and then
these insects are dissected to see the developmental stages of parasites inside the insect
vector.

Treatment-
1. Tartar emetic - 1gm (1g. dissolve in 100ml distill water, 20-35ml I/V at 7days interval for
6-8 times at weekly interval. In horse 4g/45 kg.b.wt.
2. Naganol/Suramine - 1-2/100kg. b.wt. given as 10% soln
3. Naganol and Tartar emetic- 4 injections of tartar emetic, then 3 injection of Naganol
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 given at the sesequence of NTTNTTN.
4. Antrycide methyl sulfate (Quinapyramine dimethyl sulfate) 5mg/kg b.wt. S/C as 10% soln
5. Antimosan- 3mg/kg b. wt. S/C
6. Berenil-0.8-1.6mg. It is a satative (leading drug) and it is useful in those which
are resistant to other drugs.

Control-
Use of fly repellents and insecticides and control of breeding of insect vectors.

Immunity- The most important immunological aspect is that though antibodies are produced it
does not act against the parasites due to production of relapse strain with different antigenic
characters. The antibodies produced against previous strains cannot be destroyed due to
repeated change of antigenic character.

Species of Trypanosome

2) Evansi group

a) T. evansi- It is the most prevalent species of Trypanosoma in Nepal and India is which
causes "Surra"disease, it is derived from Hindi word sudda.
Host- Horse, camels, cattle, buffaloes and dogs.

Transmission- This is mechanically transmitted by different spp. of fly belonging to the

genera Tabanus, Stomoxys, Haematobia and Lyperosia etc

Symptoms in horses
 Surra is always fatal in case of horse and death occurs between three weeks to
three months of infection.
 There is high rise of temperature about 104-106ºF
 There is watery nasal discharge seen.
 There is oedema of the limbs, chest region and sheath of the
scrotum.
 Gradual emaciation and weakness develops.
 There is conjunctivitis, paralysis of hind legs followed by death.

Symptoms in cattle and buffalo- Here four form of the disease are seen.
 Slow developing form.
 Acute and rapid developing form.
 Hyper acute type.
 Latent type

Of these four types the rapid and acute form is more common and symptoms are:
The animals are excited with high rise of temperature from 103-105ºF. Nervous symptoms
such as circulatory and striking the head to hard object. The symptoms of nervous excitement
last for 2-6 hrs. Then the animal becomes depressed and lies down. Such period of
excitement and depression last for 3days during which period the animal may die or recover.
Sometimes the animal may die without showing symptoms. The affected animal looks dull
and sleepy.

Symptoms in Dogs -
 There is rise of temperature
 The conjunctiva is congested.
 Oedema of head, throats and limbs.
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June 10, 2020 12
  Corneal opacity, keratitis, these may be partial or completely blindness.
 The pulse and respiration rate increased and death occurs within 1-3 month's time.

b) T. equinum- Morphological difference in this case is that the kinetoplast is absent. It causes
a diseased condition called "Mal de caderas" in equine. The symptoms are
a. Rise of temperature
b. Weakness of the hind legs
c. Paralysis of the hind leg followed by death
Tabanus fly mechanically transmits these parasites. The disease mostly occurs in S. America,
Argentina, Bolivia and Paraguay.

c) T. equiperdum- It is commonly known as tissue trypanosome because it is found in the

genital tissue of equine and the disease produced by this species is known as "Mal de
coitus" or "Dourine" or "Horse sleepless". It is transmitted mechanically by coitus.
Mostly found in North and South Africa, Central and South America, The Middle East and
Asiatic Russia.

3) Brucei group
1. T. brucei- Prevalent in South Africa and causes a disease condition called
"Nagana" in cattle, horse and dog and transmitted by Glossina fly.
2. T. gambiense- It is also prevalent in South Africa causes and the disease condition
called Sleeping sickness in man, sometime pigs, cattle and dog are also infected. This
parasite is transmitted by Glossina palpalis. Mostly prevalent in West Coast of Africa.
3. T. rodesiense- This parasite causes Rhodensiense disease or East African
sleeping sickness in man and trasmitted by Glossina morsitans.
4) Vivax group
1. T. vivax - The disease produced by this is also known as "Nagana". This
parasite produces chronic and acute type of Nagana in case of sheep, goat, cattle and
it is chronic in Africa and mainly transmitted by Glossina morsitans.
2. T. uniformae- It also produces the disease called Nagana in sheep and cattle.

5) Congolense group
1. T.congolense- This parasite produces acute and serious types of "Nagana"
disease in sheep, goat, cattle, horse and pig and it is found in the plasma of these
animals. It is transmitted by different species of Glossina spp.
2. T. dimorphon- The size of this parasite is about 11-24μm length and is
dimorphic, slender, has no free flagellum and the kinetoplast is marginal and
subterminal. It occurs in domesticated animals and is wildly distributed in Africa and
transmitted by Glossina spp.
3. T. simiae (syn. T. porci, T. ignotum, T. rodhaini) – These are polymorphic in
form resembling T. congolense. The majority of these parasites are 16-24μm in
length, being long and stout with a distinct undulating membrane. It is highly
pathogenic for pigs and camels. It is mostly distributed in tropical East and
Central Africa and transmitted by different spp. of Glossina.

Zoonotic Importance of Trypanosoma- The species of Trypanosoma which are found in

Nepal and India are from the Evansi group and do not have much zoonotic importance.
Though some accidental infection has taken place in man they are not of importance. Natural
transmission is not seen from animal to man. But African types of Trypanosoma are the
species that infect man such as T. gambiense and T. rhodensiense which are also found in
domesticated animals and wild animals. These animals act as a reservoir host for human
infection.

Genus Leishmania
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June 10, 2020 13
Leishmania is primarily a disease of man and dog, jackal, rodents etc. act as reservoir host.
The Leishmania strain found in Nepal and India is mainly L. donovani and it does not have a
reservoir host. These are intracellular parasites of macrophases and reticuloendothelial cells
of spleen, liver, bone marrow, lymphnodes and skin

Morphology- The parasite exists in two

forms:
1. Amastigote form: Occurs in vertebrate
host.
2. Promastigote form: Occurs in gut of sand
fly and can be cultivated in artificial
culture.

Amastigote form (A flagellar stage): The

parasite at this stage resides in the cells of the
reticulo-endothelial system of vertebrate hosts
(man, dog and rodents). The characters of the
amastigote form are as follows:
It is rounded or oval body measuring 2 to 4 µm along the longitudinal axis. Cell
membrane is delicate and can be demonstrated in fresh specimens only. Nucleus measures a
little less than 1µm in diameter. It is oval or rounded and is usually situated in the middle of
the cells or along the side of cell wall. Kinetoplast lies tangentially or at right angle to the
nucleus. It comprises a DNA containing body and a mitochondrial structure. Axonemes
(rhizoplast) are a delicate filament extending from the kinetoplast to the margin of the body. It
represents the root of the flagellum. The vacuole is clear unstained space lying alongside the
axoneme.

Promastigote form (Flagellar stage): This stage of the parasite is only encountered in
cultures and in insect vectors. The earlier stages are short oval, or pear-shaped bodies,
measuring 5 to 10 µm in length by 2 to 3 µm in breadth. The fully developed forms are long
slender, spindle shaped bodies, measuring 15 to 20 µm in length by 1 to 2 µm in breadth.
Nucleus is situated centrally. Kinetoplast lies transversely near the anterior end. Vacuole is a
light staining area lying in front of the kinetoplast over which the root of the flagellum runs.
Flagellum may be of the same length as the body or even longer, projecting from the front.
The flagellum does not curve round the body of the parasite and there is no undulating
membrane.

Life cycle- The Leishmania parasites are ingested by sand flies (Phlebotomus spp.) while
sucking blood from infected hosts. These parasites reach the midgut of the sand flies where
they change into promastigote form and the promastigote form multiplies in the midgut by
binary fission. After 6-9 days they move forward and block the pharynx of sand fly. When
these flies bite a new host these promastigote forms are injected into the blood vessels of skin.
Then these promastigote forms are engulfed by the macrophages and they change into
Leishmanial form or amastigote form and by beans of blood circulation reach various organ
of the body. Infection of animal and man is mainly by biting of infected flies. But some times
when the infected flies are crushed over broken skin; surface infection may also take place.

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June 10, 2020 14
 There are three spp. of Leishmania: donovani, L. tropica and L. brasiliensis
Infection- D. N. Banerjee (1955) has reported that congenital transmission also occurs
through blood. Symmers (1960) reports possible transmission by coitus in a curious case of
women who had never left England but who developed a leishmanial sore on the vulva; her
husband had inadequate treatment of Sudanese kala-azar some year before.
Comparative studies of three species of Leishmania
S.N . Characters L. donovani L. tropica L. brasiliensis

1 Definitive man, dog, jackal & man and dog man and forest
host rodents rodents
2 Intermediate p. argentipes, p. p. papatasi & lutzomyia spp.
host major, p. orientalis, p.sergenti
Lutzomyia spp.
3 Location macrophages & cell macrophages & endothelial & large
of the RES clasmaptocytes of mononuclear
RES, skin, cells of
endothelial cell of skin
capillaries & lymph &mucou
nodes of skin s membrane of
4 Disease visceral cautaneous nose, mouth
muco-cutaneous &
leishmaniasis leishmaniasis, delhi pharynx
leishmaniasis,
kala-azar, delhi boil, allepobutton, euspondia, uta
fever,dumdum fever, oriental sore
tropical splenomegaly

5 Incubation 3-6month,sometime few week to six a few days to few

period 1-2 yrs. month or some time weeks
1-2 yrs.

Pathogenicity
Visceral leishmaniasis caused by L. donovani- Soon after the promastigote forms are
injected in the skin capillary by the sand flies a local reaction takes place. There is infiltration
of large no. of macrophages resulting in the formation of papillae on the skin. These
macrophages engulf the promastigote form in which the Leishmanial stage is formed. They
are carried into different organs of the body such as liver, lymph gland, spleen and bone
marrow where they grow, multiply and result in impairment of their functions. From this it is
seen that Leishmania donovani or visceral leishmaniasis may result into cutaneous
leishmaniasis but not vice versa. The incubation period is about 3-6 months but it may
exceed, sometimes it takes one or two years. The main pathogenicity of Indian kala-azar is

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splenomegaly, hepatomegaly with fever and weight loss. The haemopoetic process gets
hampered leading to anaemia, leucopenia with neutropenia and thrombocytopenia.
Hypoproteinaemia, reduced immunity and muscle wasting can also be seen.

Clinical symptoms-The initial pathogenesis and symptoms are:

 Fever and headache
 Progressive enlargement of spleen and lymph gland caused lymphadenopathy
 Occasionally acute abdominal pain
 Profound anaemia with haemoglobin level 5-10gm/100ml

Later on the symptoms are:

There is marked enlargement of spleen and liver, anaemia, emaciation, oedema of skin,
diarrhoea, bleeding from the mucous membrane of gum region or epistaxis may be a
presenting symptom. If untreated 74-95% of patients die within a period of 2yrs.

Post Kala-azar Dermal Leishmaniasis (PKDL) - Post Kala-azar Dermal Leishmaniasis

(PKDL) is a morbid condition of kala-azar. This is a type of non-ulcerative cutaneous lesion
prevalent in endemic areas of kala-azar in India, Bangladesh and some part of Africa. Reports
of PKDL in China and Iraq have also been documented. It develops in about 10% of kala-
azar patients generally one or two years after completion of antimonial treatment. This
phenomenon is the result of an immune response on the part of the host that protects viscera
but not the skin, PKDL vary considerably in appearance and these lesions are of three forms.

1. Depigmented macular lesions- These are the earlier lesions. These lesions mostly occur in
the trunk and extremities but face is less commonly affected.

2. Erythematous patches- Theses are also earlier lesions which appear on the nose, cheeks
and chin. They are very photo-sensitive, becoming prominent towards the middle of the day. 3.
Yellowish pink nodules: These replace the earlier lesions and usually donot appear from the
very beginning. The lesions are found mostly on the face and skin and rarely on the
mucous membrane of the tongue and eye. Lesions are mostly found on the face.

Cutaneous Leishmaniasis: The promastigote form multiplies inside the macrophages and
changes inti the leishmanial form. They again multiply inside the macrophages resulting in
the rupture of macrophages and new macrophages that arrive at the ulcerative are also
affected. The ulcer persists for a month.

Muco-Cutaneous Leishmaniasis or Espundia -This leishmaniasis is due to L. brasiliensis. It

not seen in Nepal and India but it causes severe and horrible skin lesions in the mucous
membrane of mouth, nose and pharynx. They take considerably long time to heal up.
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June 10, 2020 16
Diagnosis-
Microscopical examination- For this purpose different material is examined.

1. Skin scrapping or smear prepared from the skin scrapping which is taken from the
periphery of the ulcer.
2. Blood smear examination- After staining with Leishmania or Giemsa staion.
3. Smear from biopsy material such as from the spleen, lymphnode, liver, iliac creast or
sternum and bone marrow and finding the Leishmania parasite.
4. After post-mortem- Impress as smear prepared from spleen, liver and bone marrow may
be examined.
5. Cultural method- Leishmania parasite can be cultured in NNN media. For this
suspected blood, biopsy material, post-mortem or skin scrapping only for L.
brasiliensis may put in the culture in NNN (Novy-MacNeal-Nickolle) media and
promastigote forms are found in the positive cases, it takes one month.
6. Biological method- The blood or skin scrapping of infected animal are injected
intraperitoneally to rat or this may be injected subcutaneously at the base of the tail and in
7. Serological test- Formal gel test or Henery's gel test- If a drop of formalin is added to a
drop of suspected serum in +ve cases gel formation takes place and whole mixture becomes
milky white in colour. It is more reliable in dog infection.
8. Chopra's et al, antimony test- About 3-4 ml suspected serum is taken in test-tube and 1-2
ml of 4% urea stilbamine is gradually added in +ve case precipitate are formed at the bottom
of the test-tube.

Treatment-
 Mapacrin-dose 10% soln of mepacrin1-2ml injected at the site of the skin lesions.
 Fuadin- dose 0.1-0.2gm in 2ml of distill water and injected I/M
 Pentamidine- dose 2mg/kg b. wt. dissolved in 2-5ml of distilled water injected I/M has
satisfactory result.
 Sodium Antimony Gluconate- 6-8gm I/m available in 20,40or 100mg/ml of
concentration.
 Amphotericin B (Fungizone) an antifungal agent @ 1mg/kg of b.wt daily by slow I/V
dissolve in 0.5 litre of 5% dextrose (given over 6hrs.). It may be increased to 0.25mg/kg b.
wt. total dose should not be more than 2gm.

Control
By controlling the sand fly or Phlebotomus flies. Once man is infected by this disease, he has
active acquired immunity developed.

ORDER-DIPLOMONADIDA

Family 3-Hexamitidae
a) Genus: Giardia lamblia
Synonym-Giardia intestinalis or Lamblia intestinalis

Location and host: The organism is found in the duodenum and other parts of the small intestine
and occasionally in the colon of man, monkeys, pigs and rats.

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Morphology: It exists in two phases i.e., trophozoite and cysts.
Trophozoite- Size of trophozoite is 14µm long by 7µm broad. The body of the parasite is pear
shaped. Anterior end is broadly rounded and posterior is pointed. The dorsal surface is
convex and ventral surface is concave. The body is bilaterally symmetrical. The anterior half of the
body has a large disk like structure known as sucking disk and these sucking discs contain two
nuclei both are vesicular type. A pair of dark staining body known as parabasal body is
present centrally. There are two axostyles and two nuclei and four pairs of flagella.

Cyst- The fully formed cyst is oval in shape and measures 12µm long by 7µm broad. The
axostyles lie more or less diagonally, forming sort of dividing line within the cyst-wall. There are four
nuclei, which may remain clustered at one end or lie in pairs at opposite poles. The remains of the
flagella and the margins of the sucking disc may be seen inside the cytoplasm.

Life cycle- Life cycle of G. lamblia is simple and completes in a single host, man. No
intermediate host is required. Infection is acquired orally by ingestion of cyst from contaminated
hand or water or food. Excystation occurs in the stomach and in the duodenum in the presence
of gastric acid, pancreatic enzymes (chymotrypsin and trypsin). Multiplication in vegetative form
is by longitudinal binary fission. Some of the trophozoites then pass down on the large intestine
where they again encyst in the presence of neutral pH and bile salts, sometime in chronic stage
of disease trophozoite reach into gallbladder. Formation of cyst begins by shortening of flagella
followed by condensation of cytoplasm and finally secretion of thick hyaline cyst wall.

Transmission - It is mainly by contaminated food and water.

Pathogenesis and symptoms- These are more pathogenic in human being and mostly
children suffer more and causes retardation of growth and so young animals suffer more
compare to adult. In chronic stage of disease it may cause cholecystopathy in infected person.

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Symptoms- Chronic diarrhoea, containing more mucous is seen. This also hampers in the fat
metabolism by forming a layer over the intestinal mucosa and due to this there is defective fat
metabolism called steatorrhea, resulting in the deficiency of fat-soluble vitamin.

Diagnosis- By finding the cyst in the faces.

Treatment- Similar to amoebiasis.

 Mepacrine (atebrin) - 200mg 30 days.
 Meteronidazole (flagyl)- 100mg thrice a day for 3-5
days.
 Chloroquine - 300mg daily for 5days in human and
pig.
 Emetine hydrogen chloride -0.5-1gm I/M or S/C injection for 10 days.
Other spp. of Giardia that are found in the domastic animal are G. canis, G. cati, G. bovis, G.
caprae, G. equi, G. muris (mouse or rat), G. duodenalis (rabbit), G. cavie (guinea pig) etc.

b) Hexamita meleagridis

Geographical distribution- It is found throughout the world and is quite common tropical
region.
Location and host- This parasite is found principally in the duodenum and small intestine of
young turkeys. It is also found in the bursa fabricious and caeca of domesticated adult birds
and wild birds like peafowl and various pheasants etc.
Morphology- The organism is bilaterally symmetrical, pear shaped and measures 6-12µm in
length by 2-5µm broad. There are six anteriorly directed and two posterior directed flagella
present. There are two nuclei and two axostyles are present. Only trophozoite stage is found.
Ant. flagella Nuclei Axostyles

Post.flagella
Life cycle- Trophozoite of parasite multiplies by longitudinal binary fission. There are no
cysts, and transmission is direct from bird to bird through contaminated feed and water.
Pathogenesis and symptoms- Young turkey up to the age of about two months are the most
susceptible. Death may occur within a week of infection in some cases, and the mortality in
the flock may reach up to 80%. There is catarrhal enteritis of the upper digestive tract and this
produces a marked lack of tone in the duodenum and jejunum. The small intestine is
inflammed and oedematous, there is congestion of the glandular tissue of the caecum. The
diseased birds also produced watery diarrhoea, later it becomes listless, loss weight rapidly
and finally die. If birds are recovered, they grow poorly and may act as carriers.
Diagnosis- This is made by the demonstration of living organisms in a drop of the contents of
the small intestine.
Treatment-
 Furazolidone: at a rate of 110mg/kg in feed could be prevented by continuous feeding.
 Nithiazide: at a concentration of 0.02% in drinking water controlled, fatal out breaks due to
Hexamita and Histomonas in turkey.
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ORDER- TRICHOMONADIDA

Family 4-Monocercomonadidae

a) Histomonas meleagridis
Location and host- It is found in caecum and liver of turkey and also sometimes in fowls. It occurs in
young between 2-12 weeks of age. It causes Histomoniasis, infectious Entero-hepatitis or 'Black-
Head,' in the turkey. It may also occur in the chicken, peafowl, guinea fowl, pheasant, partridge
and quail.

Flagellum Nucleus

Morphology- it is extracellular parasite and pleomorphic in character. It is amoeboid and

contains one or more pseudopodia. It measures 5-30µm in diameter. It has a clear and granular
endoplasm. Endoplasm contains food vacuoles with bacteria and starch granules. Nucleus is
vesicular and only one flagellum or in some there are two flagella and axostyles present. No cyst
formation takes place. It multiplies by binary fission.

Life cycle- infection can take place by simple fecal-oral transfer within a contaminated area.
Wider transmission, however, is an intricate process which starts when an intestinal histomonad
invades the tissues of another parasite co-inhabiting the caeca of the avian host. This is the
nematode worm, Heterakis (see in fig.). the protozoan travels to the ovary of the worm and is
incorporated into an egg which eventually passes into the environment with the bird’s
droppings. The worm eggs are tough and long-living. If an egg is subsequently ingested by an
earthworm, the nematode larva contained within batches and remains inside its newly adopted
transport host. Thus, the histomonad is now inside a nematode larva which, in turn, is inside and
earthworms (see in Fig.). If when the earthworm is caught and eaten by another bird, two
parasitic transmission cycles (Heterakis and Histomonas) are completed.
Dr.H.B.Rana
June 10, 2020 20
Transmission- It is by ingestion of trophozoites which are passed out in faeces.
Ingestion of infected embryonated egg of Heterakis gallinarum carries the parasite from
caecum of birds also causes infection. Incubation period is 15-20 days. Sometimes birds may be
infected after ingestion of earth worm which acts as paratenic host.
Pathogenesis - Parasite penetrates the caecal wall and causes necrotic ulcer. Lesions are mostly
found in caecum and liver. Caeca are filled with lot of exudates, which later on becomes very
hard. The liver is enlarge greatly and contains circular ulcer with a depression in the centre. The
ulcers are first yellowish green and later a fibrous tissue capsule of white colour surrounds them.
Necrotic foci may sometimes be seen in the kidney.

Symptoms- Young birds are more susceptible and adults act as carrier. Depression, dropping
wings, lowering of head and sulfur yellow colour diarrhoea is seen in birds. The disease lasts
for 3-10 days. Due to cardiac weakness the feather less part of the head has a cyanotic blue or
black colour.

Diagnosis- From the symptoms i.e. sulfur yellow coloured diarrhoea. Postmortem shows
characteristic liver ulcerations. Finding the trophozoites in faeces is also useful.

Treatment-
 Furazolidone - 0.01-0.02% concentration in food in a
day. Entramin - 0.05% as prophylactic and 0.1-0.2% as
curative  Nitrofurazone- given in food 0.1-0.4%
 NAB (Nav arseno billon) 0.1c.c./lb. b. wt. 5% soln in transmuscularly, may
be repeated once after 3 days.
 Nithiazide at a concentration of 0.02% in drinking water.

Control- Good management and hygiene.

Turkeys and chickens should be raised
separately. Young birds should be kept
separately.
Treatment of caecal wall i.e, H. gallinarum by Phenothiazine.

Parahistomonas wenrichi: The parasite is similar to Histomonas but it has four flagella
instead of one or two, an axostyle and there is rod -shaped parabasal body. This species is not
pathogenic for of gallinaceous birds. It is measures 9-27µm in diameter. It does not multiply
in the host tissues, nor does it produce any visible pathological change. The embryonated
eggs of Hererakis gallinarum transmit it.

Monocercomonas ruminantium: This paraste is found in the rumen of cattle and it is non-
pathogenic. It has only one flagellum.

Family 5 - Trichomonadidae

a) Trichomonas
The parasite consists of 4-6 flagella, out of these only one flagellum runs backward as trailing
flagellum. Only one genus is of veterinary importance in this family
Tritrichomonas foetus- Found in genital tract of cattle.
T. ruminantium- Found in the rumen of cattle and is pathogenic.
T. suis- Large trichomonas mostly found in stomach but some time nasal
passage, caecum and small intestine. It is non-pathogenic.
T. equi- It is found in caecum and colon of horses.

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 Trichomonas gallinae- Found in upper digestive tract of especially in pigeon
and sometimes found in fowls and turkey.
Tetratrichomonas gallinarum- Found in lower digestive tract and sometimes the
liver of fowl, turkey and guinea fowl.

Tritrichomanas foetus

Location and host: These are found in the vagina and uterus of cow and preputial sheath of
the bull. In case of heifer they are not found. The disease produced by this called Bovine
Trichomoniasis in which early abortion within 8-16 weeks of gestation is seen. It may also
occur in pig, horse and deer but pathogenic effects are seen only in the bovine.

Morphology- These are pear shaped with broad anterior and narrow posterior end. Size of
the parasites is 10-25µm by 3-15µm wide. The nucleus is situated at the anterior part of
the body. A parabasal body is present, which is sausage shaped or rod like in
nature. Supporting structure such as costa and axostyles are present. There are four
flagella; out of these three are anterior and 4th flagellum runs backwards along the body
of the parasite being attached to the body with the well- developed undulating membrane and
end as free posterior flagellum. At the anterior part of the body a cytostome is present.
Multiplication takes place by longitudinal binary fission. There is no cyst formation.

Transmission: Some parasites are also transmitted by coitus or copulation. A part from these
contaminated A.I. instrument, unhygienic gynaecological examination of cows acts as source
of infection and flies mechanically transmitted the parasite.

Pathogenesis and symptoms: In case of cows the parasite first invades the vagina and causes
the initial lesion as vaginitis of varying intensity. The floor of vagina may be filled with
mucopurulent discharges, these parasites further pass into the uterus through cervix and cause
endometritis and pyometra (inflammation of uterus with pus formation). The infected cow
may conceive but the parasite do not allow the foetus to grow inside the uterus this results is
early abortion. Characteristic feature of Trichomoniasis is the abortion occurring within 8-16
weeks of gestation.

Two types of abortion can be seen-

1. In first type both the foetus and foetal membrane are expelled out and not much damage is
caused to the wall of the uterus.
2. In second type only the foetus comes out but the foetal membranes are retained in the
uterus which gradually putrefy resulting in pyometra. In such case the cows usually
become sterile.
In some other cases there is death of the foetus inside the uterus, which becomes macerated
and catarrhal type of endometritis develops. In some other cases within the cervix the serous
fluid accumulates inside the uterus resulting in pendulous uterus and in such cases the fluid
that is present is grayish white or colourless and containing large number of parasites.

Dr.H.B.Rana
June 10, 2020 22
In case of bull: There is inflammation and swelling of preputial sheath and due to this there is
pain during micturition and copulation. Small reddish papillae may be present over the
prepuce and epididymis and mucopurulent discharge may be present.

Diagnosis:
 History of the herd- if there is early abortion, we can suspect this disease.
 By microscopical examination of vaginal and uterine discharge: Amniotic and
allantois fluid of freshly aborted foetus, and microscopical examination of the washing
of the prepuce of the bull. In positive case parasites are found.
 Serological type: Here one antigen known as Tricin is available and thus, 0.1ml of
1:40,000 dilution of Tricin is injected intradermally to suspected animal. In positive case a
marked swelling is seen within ten minutes of injection. It disappears within 6 hours. It is
called internal test.
 By mucous agglutination test.
 Cultural method- These parasites can be cultured in peptone broth media to which 10%
cattle serum and antibiotic like penicillin or streptomycin is added. The culture is kept at 370c
and in positive cases the parasite will grow within 28 hours.

Treatment:
For bulls- Wash the penis with a weak soln of detergent, dry it and Flavin ointment is
introduced into the preputial cavity and massaged for 15- 20 minutes. Then wash by means of
1% Acriflavine soln to kill the parasite.
For cow- Wash the vagina and uterus with 1% Acriflavine or 3% Lactic acid soln. Next
Berenil 1% soln 100-150c.c. is put into the uterus and kept these for 15-30 minutes to kill
parasites.

Control: Treatment of infected bulls, castration of infected bulls. Artificial insemination

equipment should be properly sterilized. The major control measure for trichomoniasis in
cattle is the use of A.I. in infected cow should be given breeding rest. Suspected cows should
be kept separate from other animals and without breeding.

PHYLUM- APICOMPLEXA CLASS- SPOROZOEA

SUBCLASS- COCCIDIA ORDER- EUCOCCIDIIDAE

General Characters of the class Sporozoea- The pseudopodia, flagella and cilia are absent.
There is limited motility, which is sufficient to penetrate the cell. It produces spore and
sporozoites.

Characters of the family Eimeriidae- These are intracellular parasites, found in epithelial
cell of intestine and mostly host specific.
This family has many genera, which are – Eimeria, Isospora, Wenyonella, Tyzzeria etc.

Eimeria- Sporulated oocyst; has four sporocysts and each sporocyst has two sporozoites.
Isospora- Sporulated oocyst; has two sporocysts and each sporocyst has four sporozoites.
Wenyonella- Sporulated oocyst; has four sporocyst and each sporocyst has four sporozoites.
Tyzzeria- Sporulated oocyst; has no sporocyst and oocyst consists of Eight sporocysts.

Dr.H.B.Rana
June 10, 2020 23
 Eimeria oocyst Oocyt of Isospora Oocyst of Wenyonella Oocyst of Tyzzeria

SUBORDER: EIMERIINA
Family 6 -Eimeriidae
Genus – Eimeria
Location- They are intracellular parasites and are mostly found in the epithelial cells of
intestine but some may also be found in liver and kidney of mammals and birds. Examples
are- E. tenella, E. necatrix, E. acervulina, E. maxima, E. brunetti, E. hagani, E. mitis, E.
praecox are found in the alimentary canal of poultry; E. nina, E. arlongi, E. faurei, E.
intricata etc. are found in the alimentary canal of goat and sheep. E. stiedai is found in the
liver of rabbit and E. truncata in found in the kidney of goose.
Host- All type of animals and birds act as the host of
different species of coccidia. Birds and rabbit are most
susceptible hosts; sheep, goat, cattle, buffalo, pig,
dog and cat are less susceptible hosts. Horse and
mule are rarely infected. The young animals and
chicken are most susceptible and heavily
infected.
Morphology of unsporulated oocyst- The oocyst
which has passed out freshly in faeces is known as
unsporulated oocyst. The shape is spherical, oval or
cylindrical. It is surrounded by a double layer of
oocyst wall. It is the resistant stage of the life cycle
of coccidia. There is a single mass in the centre of
the unsporulated oocyst, which is called sporont
and under favorable condition it changes into
sporulated or infective oocyst.
Morphology of sporulated oocyst- The development of oocyst outside the host body is
known as sporulation. Fully developed oocyst is known as sporulated oocyst and it is the
infective form of parasite. Sporulated oocyst of t h e genus Eimeria has four sporocysts and
each sporocyst contain two sporozoites. The anterior end has small opening called
micropyle. Micropyle is covered by a cap, which is called polar cap. In some species
micropyle and polar cap is absent. Below the micropyle there is a clear refractile granule
called polar granule. During the formation of oocyst a portion of cytoplasm is left called
oocyst residual body. The narrow end of oocyst is called stieda body. During the formation
of sporozoites some cytoplasmic mass is left which is known as sporocyst residual body. The
sporozoites are banana shaped. One end is broad and another end is narrowly pointed with a
nucleus at the middle. Cytoplasm is granular with a vacuole at each end.

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June 10, 2020 24
Life Cycle- The oocysts are passed in the faeces of host. The freshly passed ooocysts contain a
single cell; the sporont. They must have oxygen, moisture and suitable temperature in order
to develop to the infective stage. The process is known as sporulation or sporogony. The
sporont, which is diploid, undergoes reduction division. The sporont divides directly forming
four sporoblasts, each of which then develops into a sporocyst. T w o s porozoites develop
within each sporocyst. Sporulation usually takes two to four days at ordinary temperature, and
then sporonts are ready to infect the new host. Definitive host then gets infection
through contaminated food and water. In the lumen of the intestine of definitive host the
oocyst wall is ruptured in the presence of carbon dioxide, reaction of enzymes and bile juice.
Sporozoites are liberated in the small intestine. Sporozoites enter in the epithelial cells of
the intestine either directly or via white blood cells and round up to become trophozoites. The
nucleus of trophozoites divides mitotically into a large number of small bits. This
multinucleated body is called schizont. Later a portion of cytoplasm surrounds each bit of
nucleus and large numbers of merozoites are formed. When the wall of the schizont is
ruptured a large number of uninucleated first generation of merozoites are released.
The merozoites of the first generation further attack to the fresh epithelial cells and second
generation of merozoites are formed within five days after inoculation. Some of them enter
in new epithelial cells and round up to form third generation of merozoites. The number of
merozoite released from the schizont is different according to their species. In this way after
3-5 schizogonous life cycle takes place, many of the merozoites of second generation
penetrate again the fresh epithelial cell and start gametogony cycle. They produce male
and female gametocytes. The male gametocyte is called microgametocyte and the female
gametocyte is called macrogametocyte. During production of merozoites most of the
merozoites change into macrogametocytes and few merozoites are change into
microgametocytes. Later macrogamete or female gamete is formed from the
macrogametocytes and microgamete or male gamete is formed from the microgametocyes.
Microgametes are small and having flagella and many microgametes surround the single
macrogamete. In case of E. tenella only one microgamete enters inside the macrogamete
through the micropyle; the nuclei of both gametes fuse to form zygote. The cytoplasm of
zygote shrinks and become hard to form the outer wall, now the zygote is called oocyst. Inside
the oocyst the refractile granule joined together to form the inner wall of the oocyst. Oocyst
after rupturing the intestinal epithelial cell reaches in the lumen of the intestine and then
is passed out with faeces. The freshly passed oocysts are unsporulated. The sporulation of
oocyst is depend on the species and temperature and moisture of the environment. The
prepatent period of Eimeria species varies considerably and may be as short as five days in
poultry and up to 3-4 weeks in some ruminant species.

Dr.H.B.Rana
June 10, 2020 25
Mode of infection- The disease cause by the coccidia is called coccidiosis. Birds are infected
by the ingestion of sporulated oocysts of coccidia through the contamination of food and
water.

Pathogenicity and Symptoms- The clinical symptoms of coccidiosis varies with the number
of sporulated oocyst ingested and age of the birds and animals. In birds no symptoms are
observed till the 4th day of infection. There are mainly two types of pathogenicity cause by
coccidia in birds.

Caecal Coccidiosis-This type of coccidiosis is caused by E. tenella. In this case caeca

may be filled with blood tinged contents and caecal wall seems patchy or diffusely
haemorrhagic. Some times in chronic cases there may be cheesy cores in congested caeca.
In infected birds no symptoms are seen till the 4th day of infection. When the mature
merozoites are released on the 4th day of infection they pinch the capillary of caecum and
blood stained faeces pass out. Then extensive bleeding occurs on the 5th day and 80-90% of
infected birds die on 5th day. Prepatent period is seven days.

Symptoms:
 Poor growth, emaciation
 Poor egg production
 Loss of appetite
 Restlessness and drooping wings
 On the 6th day there is sluggish movement
 There is a sulphur diarrhoea
 In bird jaundice may be seen in advanced cases

In less acute case the faeces may be chocolate brown or red mixed blood.
Occasionally paralysis is seen. Mortality rate is highest between 4-6th day of infection. Death
occurs due to sever blood loss. If the birds survive the oocysts are passed out on 7th day and
birds usually recover.

Postmortem Changes- Caeca may be swollen and enlarged, the wall may be
thickened and inflammation occurs. On the 3 rd day of infection pinpoint hemorrhages may be
seen in the caeca. On the 4th day of infection a large number of haemorrhagic spots are seen
on caecal wall. On the 5th day of infection caeca is filled with large
amount of unclotted blood or partly clotted blood. The caecal content contains
schizonts and merozoites. On 7th day of infection the caeca is filled with fibrinous and necrotic
material and a large number of oocyst.

Intestinal Coccidiosis – E. necatrix causes Chronic Intestinal Coccidiosis and mostly the
middle 3rdof the small intestine is affected. The small intestine is greatly swollen with small
white small foci. Haemorrhage is seen on 5th day and lumen of intestine is filled with a large
amount of clotted or unclotted blood. In E. acervulina the anterior half of small intestine is
infected. It causes chronic diseases with mucoid diarrhoea but no blood. Intestine is
thickened with white transverse patches and the white spots take linear configuration. In case
Dr.H.B.Rana
June 10, 2020 26
of E. brunetti, the large intestine below the caecal junction and in the terminal part of the
small intestine show haemorrhagic enteritis and sometimes catarrhal enteritis.

Diagnosis- Coccidiosis can be diagnosed by the symptoms, finding the oocyst in faeces,
at postmortem from characteristic lesions on the caeca and small intestine. Intestinal and
caecal scrapping will show the presence of oocyst and other developmental stages.

Identification of oocyst- The oocyst of different species can be identified on the basis of their
shape, size, colour, location in the host type and degree of their pathogenic lesions and
sporulation time. For sporulation the caecal sample positive for coccidia parasite are kept in
0
2.3% potassium dichromate solution at 25-30 C. Sporulation takes
place within 48-72 hours.

a) E. tenella- This species is very common worldwide in the caeca of the chicken.

Oocyst- The oocysts are ovoid 14-31µ by 9-25 µ m in diameter with a smooth, two layered
wall without a micropyle or a residual body. Sporocysts are ovoid, without a residual body.
The sporulation time is 18 hours to 2 days.
Life Cycle- Prepatent period is six days. Prepatent period is about 7 days.

Pathogenesis and Symptoms- Caecal coccidiosis is found most frequently in young birds.
Chickens are most susceptible at four weeks of age. Chicks 1-2 weeks old are more resistant.
Older birds developing immunity as a result of exposure to coccidiosis due to E. tenella may
 Vary in severity from an inapparent infection to an acute, highly fatal
disease, depending on the infective dose of oocyst
 The pathogenicity of the coccidian strain
 The breed and age of the chickens
 Their states nutrition and
 Other diseases and stress often to which they are concomitantly subjected

Caecal coccidiosis in an acute disease characterised by diarrhoea and massive caecal

haemorrhages. The first signs appear when the second-generation merozoites begin to enlarge
and produce leakage of blood into caeca. Blood appears in the droppings 4DAI (Days After
Infection). The greatest amount of haemorrhage occurs in 5-6 DAI.

b) E. necatrix- This species is worldwide distributed in chicken. The first and second-
generation meronts are
produced in the small
intestine and the third
generation meronts, gamonts
and gametes are produced in
the caeca. Prepatent period is 8
days.
Small intestine
Caecum

Rectum

Oocyst- The oocysts are ovoid and 12-29µm by 11-24µm in diameter with a smooth
colourless, two layered wall without a residuum. The sporulation time is 18 hours to 2days.

Dr.H.B.Rana
June 10, 2020 27
Treatment-
 Amprolium- A combination of Amprolium and Sulphaquinoxaline at levels
of 0.006% of each in the food is more effective against poultry coccidia.
Or
 Amprolium powder or Amprosol 1gm dissolved in 1litre of water for 5-7 days.
 Coxiquin or Duoxoxin- 1gm powder dissolved in two litres of water for 5 days.
 Codrinal powder- 4gm dissolved in 1litre of water for3-6 days.
 Esb3 powder- 1gm powder dissolves in 1litre of water and for 3-6 days.
Sulphaclozine or sulphaclorpyrazine.
 Duocoxin- 30gm/ 30lit water provided for 3 days.
 30gm/ 50lit water for 3 days.
 Coccinil- 30gm/30lit water for 3 days.
 Coximar- 30gm/30lit water for 3days.
 Supercox- 1gm/1lit water for 3days (Only pure water for two days.)
1gm/2 lit of water for 3days. = Total eight days should be treated.
 Other coccidiostats are: Coxistac, Sacox-120, Meridot, Cocciwin, DOT
Maduramycin etc.
 Supplements-Vitamin K containing Kaysol Forte 5gm for 200 birds,
liquid 5ml/100birds or 10ml/100birds provides for 3-5 days in water.
 Vita Blend WM Forte (Vit.A) 2ml/100 birds provided continuously for 3-5
days, it helps in the regeneration of epithelial tissues. But we should not use Vit.
B complex since it contains Thiamin which promotes the growth of the parasite.

Prevention and Control Measures of Poultry Coccidiosis-

1. Destruction of oocyst- The oocysts can be killed by the following methods.
a. Fumigation of poultry houses and litters by 0.044% solution of ammonia. This
will kill the oocyst with 2 hours.
b. By application methylbromide at the rate of 1lbs/1000sq. ft of poultry litter.
2. Proper sanitation and management should be adopted in poultry houses.
3. Young birds should be kept separately from adult ones.
4. The poultry litters should be kept dry and stirred frequently or it should be
changed frequently when it becomes wet.
5. In case of out breaks, sick birds should be removed immediately and kept in a
separate house. The remaining healthy birds should be treated with coccidiostats for
preventing the disease and the infected litter should be changed.
6. As the oocyst can be carried by cloths of attendants of the poultry house the shoes
should be removed outside the poultry house before entering into it.
7. In some places vaccines are used against coccidiosis. These vaccines are helpful
by immunizing the chick. The vaccines are prepared from a mixture of four
common pathogenic species such as E. tenella, E. necatrix, E. acervulum and E
.maxima, sometime also for E. hagani. These oocysts mixture is given to 3-5 days
old chick in a small dose mixed with the feed. Along with this oocyst some
coccidiostats are also given to check the clinical sign of infection.

COCCIDIOSIS IN RABBITS
The species of Eimeria found in rabbit are:
 E. stiedai – Most pathogenic coccidiosis of rabbit found in liver and bile duct.
 E . megna
 E. perforans
 E. irresidua
These all are found in the intestine of rabbit.
a) Eimeria stiedai

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June 10, 2020 28
Geographical Distribution- It is cosmopolitan in distribution.
Location and Host- Parasites occur in the liver and bile duct of rabbits. This is the most
common pathogenic coccidium of domestic rabbits.

Sporulated oocyst of E. stiedae

Morphology- The unsporulated oocyst is ovoid and sometimes ellipsoidal with a flat
micropylar end, being smooth and measures about 26-40µm by 16-25µm in diameter.
Life Cycle- The meront (Schizont) are in the epithelial cells of the bile ducts. The number of
asexual generations is unknown but is apparently indefinite. At least five or six are known and
most of them have two types of meront (Schizont). One type of meront produces a small
number of plump merozoites and other type produces numerous slender ones. The prepatent
period is unknown but it may apparently be several weeks.
Pathogenicity and Symptoms- In mild infections little or no clinical signs are evident but in
heavy infections severe liver involvement is seen. Liver is greatly enlarged with many white
spots (necrotic foci). In heavy infection diarrhoea and death may occur. In fatal cases the
liver may be five to ten times its normal size. This is also associated with Ascites sometimes.
Petechial haemorrhages are seen in liver and kidneys. Oedema is seen in the peritoneal cavity
and the whole body may be oedematous. If the animal survives, an invasion of the lesions by
fibrous tissue occurs, new bile duct proliferates, and large area of the liver is transformed into
fibrous tissue. E. steidai is a scourage of rabbits and is difficult to eradicate from the colonies.
Treatment- Sodium sulphamezathine – 0.2% in water for 6-7 days highly effective
Sulphaquinoxaline – 0.03% in feed for 6-7 days
Sulphaguanidine – 0.5% in feed for 6-7 days
The intestinal forms cause loss of appetite, diarrhoea and rough hair coat. Coccidiosis is a
major problem in rabbits and it is one of the major causes of deaths in rabbits as well.

COCCIDIOSIS IN SHEEP AND GOAT Various species of Eimeria are:

 E. arlongi
 E. nina = Kholyokimovae
 E. faurei
 E. intricata
E. arlongi is the most pathogenic one of these species of Eimeria. Young animals are mostly
affected and symptoms include restlessness, weakness, abdominal pain, lost of appetite,
diarrhoea, which may be foul and faeces mixed with blood and mucous is also seen.

COCCIDIOSIS IN CATTLE AND BUFFALO

The species of Eimeria found in cattle and buffalo are:
 E. zuernii
 E. bovis
 E. cylindrica
 E. subsperica

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June 10, 2020 29
a) E. zuernii
Geographical distribution- Worldwide in distribution.
Location and host- Schizogony stage of parasites are mostly found in the small intestine and
gametogogy stage of the parasite is found in the caecum, colon and rectum of the cattle and
buffalo. This species of the coccidium is the most common and most pathogenic ones.

Unsporulated oocyst of E. zuernii

Morphology- The unsporulated oocyst is spherical, sub-spherical to ellipsoidal. The sizes the
oocysts are 15-22µm by 13-15µm in diameter. The oocyst wall is thin, and
colourless to pale yellow in colour. The micropyle is not located. Sporulation time is 3days at
200 C and 23-24 hours at 30-32.50C.

Life cycle- By the second and third days of infection, trophozoites are found in the mucosa,
some penetrating as far as the muscularis mucosa. By the sixth day schizonts are found in the
epithelial cells of the upper and lower parts of the small intestine. Schizonts may still be
present up to the nineteenth day and by this time parasites occur throughout the small
Dr.H.B.Rana
June 10, 2020 30
intestine and also in the caecum and colon. Mature schizonts measure up to 7-9.8µm in
diameter and it produces 24-26 merozoites. There is two generation of schizogony the first
occuring in the lower ileum and the second in the colon and caecum. The earliest sexual stage
is the macrogamete and it seen 12 days after infection in the epithelial cells of the villi of the
lower small intestine and in the caecum, colon and rectum. The microgamonts are seen first
on the fifteenth day, being found in the lower colon and rectum. They are much fewer than
the macrogamonts. Oocysts may be found in the tissues of the caecum and colon as early as
12 days after infection but oocyst production is highest at 19-20 days after infection.

Pathogenicity and symptoms- E. zuernii is the major pathogenic coccidium of cattle and
buffalo. Haemorrhagic diarrhoea, anaemia, weakness and emaciation are characteristic
features of acute diseases. In severe infections death may occur as early as seven days after
the one set of clinical signs. At postmortem the major lesions occur in the large intestine,
although gerneral catarrhal enteritis may be present in both the small and large intestine.
Smear from the mucosa show very large number of developmental stages of oocysts.

Diagnosis- This disease is diagnosed by the finding of oocysts of parasite in the faecal
examination.

Treatment- Amprolium- 20-30mg/kg b. wt. daily in feed for 4to 5 days.

Lincomycin hydrochloride- 1gm/calf given in the drinking water for 21 days.
Preventive measures- The prevention of bovine coccidiosis is based on treatment and good
sanitation.

b) E. bovis

Geographical distribution- Worldwide in distribution.

Location and host- Schizogony stages of this parasite occurs mostly in the small intestine
and the sexual stages in the caecum, colon and terminal part of the ileum of ox, zebu and
water buffalo.
Morphology- Unsporulated oocysts are ovoid, smooth and average size is 27-29µm by 20-
21µm in diameter.
Life Cycle- There is two asexual generations. The mature first generations of meronts are
about 281µm by303µm in diameter. It is the largest schizont among the all species of Eimeria
and contains an average of 120,000 merozoites in each. They are easily visible to the naked
eye as whitish balls. They lie in the endothelial cells of the lacteals within the villi in the
posterior half of the small intestine. The second generation of meronts or schizonts is in the
epithelial cells of the villi of the caecum and colon. The average size is 10µm by 9µm in
diameter in tissue sections and contains 30 to 36 merozoites in each. The gamonts occur in
the epithelial cells of the intestinal villi, many in the caecum and colon, but may extend into
the small intestine in heavy infections. The prepatent period is 15 to 20 days and the patent
period is 5 to 26 days.
Pathogenisis and Symptoms- E. bovis is probably the most common cause of coccidiosis in
cattle. In severe infections majority of the crypts of the large intestine and sometimes the
terminal part of small intestine are destroyed and the lumen of intestine is filled with blood.
The mucosa is necrotic and this damage may extend to the submucosa. The wall of the intestine
is congested and oedematous.
Diagnosis- By finding oocysts in faecal sample and from the symptoms of the disease.
Treatment- Amprolium 10mg/kg b.wt. for one week.
Decoquinate 0.5mg/kg b. wt., at least 24 days for protection.

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June 10, 2020 31
PHYLUM: APICOMPLEXA CLASS: CONOIDASIDA
SUBCLASS: COCCIDIASINA
ORDER: EUCOCCIDIA

Family 7 : Cryptosporidiidae
Genus: Cryptosporidium
Species: C. parvum
Cryptosporidium is a protozoan organism which causes the parasitic infection,
cryptosporidiosis. It exists in either the free-swimming (trophozoite) form or the oocyst
(dormant) form. Cryptosporidium parvum is now recognized as a human pathogen which can cause
severe diarrheal illness.
Geographical distribution- Worldwide.
Location and host- Trophozoite stage are found in hat are located along the gastrointestinal
epithelium, specifically the small intestine cattle, sheep and pigs, wild animals and man.

Oocyst of Cryptosporium parvum (A, 400x and B,1000x) Sporulated oocysts of C. parvum

Morphology- The oocyst of parasite is 3- to 4µm in diameter, which is spherical structures. At light
microscopic levels, the spherical structures are often seen attached to the epithelial cell surface. The
attachment site usually lacks microvilli and is dense and raised. Sporulated oocyst has four sporozoites.
Lifecycle- Lifecycle is completed in only one host. Infection occurs with the ingestion of
viable Cryptosporidium oocysts of contaminated drinking water, the environmentally
resistant form of the organism, is the major mode of transmission. Cryptosporidium
is introduced into the water supply via animal excreta containing oocysts. Important
reservoirs of the oocysts include cattle, sheep and pigs. Cryptosporidium is also found in
wild animal populations. The organism is more prevalent in wild ruminants such as deer, elk,
moose and caribou and is primarily found in neonates of these species. Person to person
transmission is common, especially in child daycare settings. Direct contact with infected
animals, especially calves and lambs, can cause illness in exposed persons. Contaminated
food can also cause infections.
The Cryptosporidium life cycle is complicated and fascinating. For the unfortunate person
(often called the host) who contracts this parasitic disease, it begins with swallowing a
microscopic Cryptosporidium parvum oocyst.
 Oocysts, found in untreated surface water, contaminated water supplies, cattle
manure, and on objects contaminated with human feces, pass through the stomach and enter the
small intestine.
 Induced by the favorable environment in the intestine, oocysts open (excyst), each one
releasing four crescents shaped sporozoites. Each sporozoite is a tiny parasite.
 The sporozoites push through the surface of intestinal cells, but they don’t go all the way
inside. They stop between the layers of the cell wall and begin to multiply, producing
the next parasitic stage six to eight merozoites. This is asexual multiplication: male
and female parasites don’t exist at this stage.
 Merozoites break out of the host’s cells, invade new cells and repeat the process.
Dr.H.B.Rana
June 10, 2020 32
 Merozoites can continue to invade new cells and multiply in a repeating cycle that result
in vast numbers of Cryptosporidium parvum parasites and infested intestinal cells.
Eventually, however, some merozoites develop differently and become male or female
gametocytes.
 Male and female gametocytes join to form gametes, developing between the layers of the
cell wall as before. They become oocysts.
 Oocysts break out and are carried away with the intestinal contents. At this stage, the host
is likely to have symptoms of cryptosporidiosis—diarrhea quickly carries oocysts out of the
body.
 Some oocysts mature very quickly and are able to release their sporozoites before they
are passed in the stool. These sporozoites immediately invade intestinal cells and begin to
multiply, starting the process of Cryptosporidium infection over again.
 When the host’s immune system is healthy, the body eventually fights off the parasite, the
intestine heals and symptoms subside. For those whose immune system can’t kill off the
parasite, Cryptosporidium infection can be virtually endless.
 Oocysts passed into the environment are infective and resilient. They are unaffected by
cold, chlorine, and other chemicals added to water to disinfect it. They survive a long time
in moist conditions, but perish if they dry out. If they are swallowed by another suitable
host, a new Cryptosporidium infection ensues.
There are several stages in the Cryptosporidium life cycle where the parasite multiplies
within the host: when the original sporozoites multiply asexually, when the merozoites
continue asexual multiplication, and when oocysts mature quickly and excyst releasing
more sporozoites. This is why a symptomatic Cryptosporidium infection can develop
after a person swallows just a handful of oocysts. A person with cryptosporidiosis passes
millions of infective Cryptosporidium parvum oocysts.

Potential Health Hazards: Development of Cryptosporidium occurs more rapidly than

many textbooks imply, and each generation can develop and mature in as little as 12-14
hours. Due to the rapidity of the life cycle, plus the auto-infective cycles, huge numbers of
organisms can colonize the intestinal tract in several days. The ileum soon becomes crowded
and secondary sites are often infected, such as the duodenum and large intestine. In
immunosuppressed individuals, parasites can sometimes be found in the stomach, biliary and
pancreatic ducts, and respiratory tract. Diarrhea, weight loss, and abdominal cramping are
clinical signs of the disease and in immunosuppressed individuals electrolyte imbalance may
occur.

Dr.H.B.Rana
June 10, 2020 33
The prepatent period is generally 4 days (3 days in heavy infections). Patency, which is the
length of time oocysts are shed in the feces, generally lasts 6-12 days in immunocompetent
individuals but may be prolonged in immunosuppressed patients. Infections are most
common in young children and immunocompromised individuals. (AIDS/HIV patients,
chemotherapy patients and organ transplant patients.) Symptoms of cryptosporidiosis usually
appear within two to ten days after ingestion of the parasite. Symptoms include watery
diarrhea, headache, abdominal cramps, nausea, vomiting and lowgrade fever. These
symptoms may lead to weight loss and dehydration. In otherwise healthy individuals,
symptoms usually last from one to two weeks, at which time the immune system is able to
stop the infection. In the immunocompromised, the infection may continue and become life-
threatening. At present, there is no effective drug therapy for cryptosporidiosis.

Treatment: Water treatment for Cryptosporidium relies on properly designed and operated
filtration systems. Chlorine disinfection of the organism is ineffective, as it has been shown
that even one oocyst can withstand pure bleach (50,000 ppm chlorine) for 24 hours and still
cause an infection. Filter systems usually consist of several filters. A "roughing filter"
containing a 5µ - 10µ (micron) cartridge filter is installed to remove any large diameter
sediments, such as iron sediments, sand, salt, etc. Downstream from the roughing filter, a
"polishing filter" containing a <1µ absolute cartridge filter is installed to remove small
particles including Cryptosporidium, from the water. Most reputable water system vendors
are currently recommending a filter porosity of <1µ to submicron or membrane filters to
remove Cryptosporidium cysts and trophozoites from drinking water.

Supportive therapy such as IV fluids is the primary for C. parvum infection. Paromomycin
and Nitazoxanide may alleviate some of the diarrheal symptoms, however the latter is
contraindicated for AIDS patients. Continuing antiretroviral drugs to boost the immune
system may also control infection. Research into other potential drugs and therapeutics
targets, as well as vaccine candidates, is ongoing. Spiramycin can be used for
immunosupressed patients.

FAMILY 8–SARCOCYSTIDA
Sub families -
1. Toxoplasmatinae 2. Sarcocystinae
Genera a. Toxoplasma a. Sarcocystis
b. Besnoitia
c. Hammondia

Characters of the sub family 1- Toxoplasmatinae

 Oocyst with two sporocysts, each sporocyst with four sporozoites
 Parasites are facultatively or obligatory and heteroxenous
 Definitive hosts are feline animals
 Merogony in both intermediate and final hosts and infection may occur
in intermediate and final hosts
 Metrocytes are not formed
 Schizonts and gametocytes are found in the enteric cells of felid and
sporogony is occurs in the outsides of the host body.
a) Toxoplasma gondii
Geographical distribution- Cosmopolitan in distribution.
Location and Host – Definitive host is domestic cat and various wild feline
species. About 300 species of warm-blooded animals such as mammals and birds
act as intermediate host.

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June 10, 2020 34
 Usporulated oocyst Sporulated oocyst of T. gondii Sporozoite stage of T. gondii
Morphology – Oocysts has two sporocysts and each sporocyst consists of four sporozoites.
The size of oocysts is about 12µm by 10µm in diameter and size of sporulated
oocyst is about 13µm by 10µm in diameter. The size of sporocyst is about 8.5µm by 6µm in
diameter and the size of sporozoite is 8µm by 2µm in diameter.
Life Cycle- Toxoplasma gondii has two developmental cycles i.e. Enteroepithelial cycle and
Extra-intestinal cycle.

Enteroepithelial Cycle – The enteroepithelial cycle occurs in cat. Cat may be infected by
sporulated oocyst or by eating Bradyzoites or Tachyzoites containing tissue of the infected
intermediate host. Kittens are infected with cysts containing Bradyzoite derived from mice.
Bradyzoites enter intestinal epithelial cells and a number of morphological types of
multiplicative stages occur. Multiplicative stages are divided into A, B, C, D, and E. Type A is
the smallest multiplicative type and it appears 12-18 hours after infection. Division of A
type is by endodyogeny and this type changes into type B and it occurs 12-34 hours after
infection. Type B has a centrally located nucleus and it divides by endodyogeny and
endopolygeny and it forms C type and it occurs 24-54 hours after infection. Type C divides by
schizogony and forms type D and it occurs from 32 hours to 15 days after infection. Type D is
the smaller than type C, it divided by endodyogeny, schizogony and produces type E, which
occurs 3-15 days after infection. Type E is multiplied by schizogony and it resembles with
type D. Later on, type E produces male and female gametes by the process of
gametogony. These gamonts occur throughout the small intestine and are common in the
ileum 3-15 days after infection. Oocyst formation occurs in the epithelial cells of the small
intestine after fusion of male and female gametes. Oocysts are discharged from the epithelial
cells and expelled out with faeces. In this case the prepatent period is three to five days and
peak oocyst production occurs between five to eight days. After feeding on sporulated
oocysts the prepatent period in cats is 21-24 days and after feeding tissue containing
tachyzoites is 9-11 days.

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Extra-intestinal Cycle- This type of life cycle mostly occurs in extra-intestinal tissue of
nonfeline species like mammals and avian host but they may also occur in the cat. The extra-
intestinal life cycle in intermediate host may start after the ingestion of sporulated oocyst or
by tissue containing Bradyzoites or Tachyzoites stages of parasites. Later different organs
of the intermediate host are infected via blood circulation. In the definitive host, extra-
intestinal cycle may start almost simultaneously with the entero-epithelial cycle of
development. There are two developmental stages found in extraintestinal development:
a. Tachyzoite or Endozoites in a rapidly multiplying form during acute infection.
b. Bradyzoites or Cystozoites in a slow multiplying encysted form, which is seen in chronic
infection.

Tachyzoite or Endozoite formation- This type of infection is seen especially in acute

visceral infection. In cats this type of development occurs in mesenteric lymph nodes and
other organs in which there is possibility of coexistence with the enteroepithelial cycle. In other
animals Tachyzoites infection occurs by ingestion of sporulated oocyst or infected meat of
another intermediate host. It develops in a vacuole in various types of cells, including
Fibroblasts, Hepatocytes, Reticular cells and Myocardial cells. In this case organisms
multiply by endodyogeny. About 8-16 organisms accumulate in a host cell after division.
Later on, they infect to new cells. The accumulation of Tachyzoites in cells is called terminal
colonies aggregates or Pseudocysts.

Bradyzoite or Cystozoite formation- Bradyzoites contained in cysts are characteristic of

chronic infection and occur mainly in the Brain, Heart and Skeletal muscles. Bradyzoites
multiply slowly by intracellular endodyogeny. Cysts may measure up to 100µm in diameter
and contain up to 60,000 organisms. Bradyzoites resist peptic and tryptic digestion. Oocyst,
which are infected from oral route or subcutaneously or intra peritoneally are more infective
than Tachyzoites and Bradyzoites in cysts.

Toxoplasmosis in Cats - Cats play an important role in the epidemiology of Toxoplamosis.

Generally, cats are commonly infected in nature and the parasite produces millions of oocysts
but rarely produces clinical disease. In cats Toxoplasmosis causes enteritis, enlargement of
mesenteric lymph nodes, pneumonia, perivascular and degenerative changes in the central
nervous system, encephalitis, diarrhoea etc.

Toxoplasmosis in Dogs – In dog toxoplasmosis causes fever, anaemia, respiratory distress

and haemorrhagic diarrhoea etc. The main pathogenic lesions are necrotic nodules produced in
the parenchymatous tissue of lungs. The spleen and liver are usually enlarged and the
organisms can be found in the liver. Ulcers in the digestive tract can also be seen.

Toxoplasmosis in Cattle – The clinical signs in cattle are dyspnea, coughing, sneezing,
nasal discharge, trembling, shaking of the head and rise of the temperature is also seen in
several cases. The main pathogenesis in cattle is fibrinous deposition in the peritoneal cavity,
enlargement of sub maxillary and branchial lymph glands, haemorrhagic tracheitis and
pneumonia with consolidation and in chronic form the blood vessel wall is calcified.

Toxoplasmosis in Sheep – The main symptoms of the disease in sheep is abortion. The
severity of congenital infection depends on the duration of gestation at time of infection.
Infection early in gestation (e.g. 45-55days) causes death of the foetus and infection at a later
time of gestation (90 days) causes other serious symptoms of the disease. Infection at 120
days of gestation causes faetal infection but death does not occur.

Toxoplasmosis in Pigs – Usually young pigs are affected by this disease and symptoms are
fever, shivering, weakness, coughing, relaxation of abdominal muscles and diarrhoea. The

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June 10, 2020 36
most common post-mortem signs are pneumonia, necrosis of the liver cell, hydrothorax,
ascites, lymphadenitis, enteritis etc.

Toxoplasmosis in Birds – A large number of records have been made concerning the
prevalence of Toxoplasma infection in birds. In fowls the symptoms are anorexia,
emaciation, diarrhoea, blindness, sometime death also occurs without any evidence of
previous illness. Histopathological symptoms show pericarditis, diffused myocarditis, a
necrotic hepatitis and also ulcers are found in the gastrointestinal tract, sometime active
follicles develop in the Bursa Fabricio’s.

Toxoplasmosis in Man – The common symptoms of Toxoplasmosis are abortion. Most of the
cases of Toxoplasmosis in children are congenital in origin; the mother usually shows a mild
infection or no infection. Maternal to fetal transmission of parasites occur less commonly
early in pregnancy than later but the severity of the damage to the fetus is greater with early
infection than later. Severe infections acquired early in pregnancy causes abortion. In less
severe infections the pathogenesis is mostly found in central nervous system than in the
visceral and somatic tissues. The main lesions are cerebral calcification, choroido-retinitis,
hydrocephalus and psychomotor disturbances. The child may be either alive or dead, and if
born alive may suffer serious mental retardation within a few weeks of birth. Acquired
toxoplasmosis (i.e., non-congenital) is suspected when lymphadenopathy, lassitude
accompanied by fever, lymphocytosis, eye lesions of doubtful origin or myocarditis are
observed. The transmission from infected meat is very rare.

Public Health Significance of Toxoplasmosis – Transmission by cats is an important factor

in the epidemiology of the infection. The role of non- cat transmission (e.g., by meat) has not
yet been established. In Britain a study showed that the general prevalence of toxoplasmosis
was 25% in the population. The prevalence increases up to the age of 20 years. Preventive
aspects for pregnant women should be to stop playing with cat, washing of hands before
eating; gloves should be worn when gardening and uncooked meat should not be fed to cats.

Treatment- No completely satisfactory treatment for toxoplasmosis is known

1. A combination of Sulphadiazine (120mg/kg b. wt.) and Pyrimethanmine (1mg/kg b. wt.)
given orally has reduce the oocyst shedding of Toxoplasma in cat.
2. Intramuscular injection of 2mg/kg bd. wt. Pyrimethamine with 100mg/kg b. wt.
Sulphadiazine inhibits oocyst shedding.

Genus- Besnoitia
Species- besnoiti
Geographical distribution- Besnoitiosis is epizootic in the south of France, but is now widely
distributed in Africa, Asia and in Southwestern Europe.
Location and host- Besnoitia besnoiti is the causative agent of bovine besnoitiosis acts as intermediate
host, is an obligate intracellular protozoan. The disease affects mainly young cattle and the final host is
cat.
Morphology- The size of the oocysts measures 14-16µm by 12-14µm in diameter and developed
intestine of cat. In bovine the parasite is found in the dermis, subcutaneous tissue and fascia, larynx,
nasal and other mucosae. The Besnoitia cyst may reach up to 600µm in diameter. Mature cysts packed
with crescent trophozoites called bradyzoites.
Lifecycle- Lifecycle completed in two hosts bovine are infected after ingestion of sporulated oocysts.
Trophozoites (bradyzoites) appears after about 16-18 days of infection and found extracellularly or in
monocytes in the blood. The most likely pathway of transmission would be transcutaneous, by stinging
insects (tabanids, stomoxys) in case of bovine. Final host is infected by ingestion of bradyzoite or
tachyzoite stage containing muscles of infected cattle. In cate parasite is developed in intestinal epithelial

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June 10, 2020 37
cell by schizogony process of multiplication then oocysts are formed. Later, oocyst infect the cattle and
the lifecycle is repeated.

Pathogenesis and symptoms- The mortality of infection is usually below 10%, although animals
may lose condition markedly, pregnant animals may abort, bulls may become sterile and the hides are of
little value for leather making purposes. During infection, an incubation stage of 3 to 6 days is followed
by 3 successive clinical stages:

- A febrile stage of 3 to 7 days, while the tachyzoites multiplication in endothelial cells of blood vessels
increases the animal temperature

- A second stage of 1 to 2 weeks, while the bradyzoites cysts generate subcutaneous oedema.
- A chronic stage of several months, characterized by alopecia and scleroderma. The skin becomes then
markedly thickened and wrinkled, and parasitic cysts are observed on conjunctiva and sclera. This
ultimate phase leads generally to the death of the animal or to its euthanasia.

Diagnosis- Serologic tests are available for the detection of the specific antibodies of Besnoitia
besnoiti present in the chronic stage. However, in order to avoid contaminated animals transfers and
control the spread of the bovine besnoitiosis in France, the diagnostic tools have to detect the pathogens
at the early stages of the disease.
PCR tests are able to detect the parasites by the detection of tachyzoites of Besnoitia besnoiti during
febrile stage in the monocytes of blood and in the skin during oedema and chronic stages.
Treatment- Decoquinate= 0.5mg/kg b. wt., at least 24 days for protection.
Sulphadiazine= 10mg/ml I. m for 5 days
Diclazurilay , Imidocarb, my also try.

Genus-Hammondia hammondai
Geographical distribution- Worldwide.
Location and host- Definitive host is cat and parasite multiply in intestinal wall, tachyzoite or cyst
stage found in skeletal muscle, heart muscle and brain.
Morphology- Sporulated oocysts has two sporocysts and each sporocyst has four sporozoites.
Tachyzoite or cystic stage is found in rat, dogs, guinea pigs and hamsters.
Lifecycle- Lifecycle completed into two hosts. Intermediate hosts are either infected by ingestion of
oocyst or ingestion of infected muscle of others intermediate host. The final host is usually infected by
ingestion of infected muscle of intermediate host. In cat, after ingestion of tachyzoite the parasite
multiply in the lamina propria of intestine, peyer’s patches and the musculature. Necrosis of infected
cells occurs and cysts appear after two weeks of infection.
Pathogenesis and symptoms- In cats Toxoplasmosis causes enteritis, enlargement of
mesenteric lymph nodes, diarrhoea etc. in intermediate host the main pathogenesis in cattle is
fibrinous deposition in the peritoneal cavity, enlargement of sub maxillary and branchial
lymph glands, haemorrhagic tracheitis and pneumonia with consolidation and in chronic
form the blood vessel wall is calcified.
Diagnosis- By finding oocysts in faecal examination of final host. Serological examination of
intermediate and final hosts.
Treatment- Same as Besnoitia.

Genus- Neospora caninum

It is a recently recognized protozoal infection of dogs and experimentally has been induced in
rodents and cats. Neospora caninum is an obligate intracellular parasite was confused
previously with T. gondii. Only sexual stages are known and thus they resemble with T.
gondii. The complete life cycle on N caninum is unknown, but it can be transmitted
trans placentally in dogs and subsequent litters may be affected.

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The tachyzoites are 5-7 by 1-5µm in diameter, depending on the stage of division. They
divide by endodyogeny. Tachyzoites are found in myocytes, neural cells, dermal cells,
macrophages and other cells. They are often located directly on the host cell cytoplasm
without vacuole. Tissue cyst up to 10µm in diameter are found in neural cells, the cyst wall is
amorphous and up to 4µm thick. Cysts have no septa, and enclose slender 7 by 1.5µm in
diameter of the bradyzoites.

Clinical Findings – Both pups and older dogs are affected. Not all littermates are affected.
Most severe infections are in young pups and typically evident as ascending paralysis of
limbs particularly hind limbs. The paralysis is often progressive and results in rigid
contraction of the muscles of affected limbs. In some dogs, only neural signs are observed.
The syndrome of Polyradiculoneuromyositis appears typical of Neosposis. Ulcerative
dermatitis, hepatitis, pneumonia and encephalitis may also occur.

Lesions – Non suppurative encephalomyelitis, polyradiculoneuritis, acute necrotizing

myositis, phlebitis, multifocal/coagulative hepatic necrosis, and atrophy of muscles are
predominant findings.
Diagnosis – Occasionally parasites are observed in biopsy specemens. An immunoperoxidase
test using specific antibodies can identify N. caninum in tissue section of biopsy smears, and
indirect FA test can be used to detect antibodies.
Treatment – Treatment is unknown but drugs used to treat Toxoplasmosis (sulphadiazine,
daraprim, scindamycin) can be tried.

Subfamily- Sarcocystinae
Genus- Sarcocystis cruzi

Location and Host- Definite hosts are dog, cat, wolves, foxes and hyenas and intermediate
host are cattle, sheep, goat, pig and man. Parasites are found in the submucosa of intestine in
definitive host and striated and cardiac muscles of intermediate host.

Morphology- The shape of the oocyst is rounded and sporulated oocyst consists two
sporocysts and each sporocyst consists four sporozoites. Size of the sporocyst is about 14-
17µm by 9-13µm in diameter. Sporozoites are banana shaped and measure about 10µm in
length. The size of the cysts is about 25-35cm in diameter which is usually found in striated
and cardiac muscle of intermediate host.

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Life Cycle- Sarcocystis has adapted its life cycle completely to the predator-prey relationship
existing between its two hosts. The prey host cattle is infected by ingestion of sporulated
oocyst from faeces of the predator like dog. The intermediate host is infected with
sporozoites; first multiplication takes place by schizogony in vascular endothelial cells of
kidney, brain and striated muscle which can be seen one month after infection. Muscle cysts
are formed in striated or cardiac muscle which contains metrocytes. The parasites occur in
elongated cysts such that may be either so small that they can’t be seen without a
magnifying lens or so large that the size reaches up to 5cm in diameter. The motile
merozoites or bradyzoites are formed by metrocytes and are present inside the cyst after
two and half to three months. After eating the cyst containing meat of intermediate host
the final host is infected. In the final host the merozoites changes directly into macro and
microgametes and start gametogony and sporogony below the epithelium of the gut. Contrary to
coccidias, no schizogonic multiplication occurs in the gut wall of the final host. Mortality
occurs 33 days after infection by feeding 105 -10 6 of sporocysts. Sporozoite formation takes
place in the gut and results in the formation of oocysts sporocysts containing two each
with four sporozoites. These sporulated sporocysts are eliminated with the host faeces. Life
cycle of S. cruzi in dog, wolves etc. shows that they shed sporulated sporozoites in faeces.
Schizonts occur in the endothelial cells of blood vessels, especially the kidney and brain.
Later cysts (metrocysts and bradyzoites) are formed in the muscles.

Pathogenicity and Symptoms- Abortion is a common phenomenon in cattle; anorexia,

pyrexia, anaemia and loss of weight, reduced milk yield, dyspnea, chronic illness, wasting
characterized by emaciation, submandibular oedema etc is seen. At autopsy haemorrhages in
various organs including the myocardium can be seen. At postmortem a generalized
lymphadenopathy is also observed.

Diagnosis- By finding sporulated oocysts in faeces of definitive hosts.

Other species of Sarcocystis

1. Sarcocystis in sheep- Sarcocystis ovicanis- Definitive host is dog and intermediate host
is sheep. This parasite is highly pathogenic for lambs which become anorectic, weak and
may die. Pregnant ewes may abort in some cases when infected by this parasite. Other
symptoms are anaemia and loss of weight, the heart is the most severely affected organ.

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This parasite is not found in the foetus, placenta or uterine tissue and this indicates that the
intrauterine infection is rare.
2. Sarcocystis in swine- Sarcocystis porcifelis-This parasite consist of a cat-pig cycle and
it is pathogenic for swine. The prepatent period is five to ten days.
3. Sarcocystis of man- Sarcocystis hominis and Sarcocystis porcihominis- Definitive host
is man and intermediate host is pig. Man is infected by the infected flesh of pig. The main
symptoms of this disease are anorexia, nausea and diarrhea.

Family 9 – Haemogregarinidae

Organisms of this family are similar to Eimeridae, but micro- and macrogamonts become
attached to each other in pairs (syzygy) during development into gametes; the zygote
becomes an oocyst producing numerous sporoblasts, each of which develops into a spore
containing two or four sporozoites. Parasites occur in the cells of circulatory system.

a) Hepatozoon canis
Hosts include dog, cat, jackal and hyena. Schizonts occur in the endothelial cells of spleen,
liver and bone marrow as round or oval bodies, more or less filling the host cell and
containing 30-40 nuclei. Various types of schizonts have been described; one produces a
small number of large merozoites (usually three) which Weyon (1926) considered became
schizonts, and another produces a large number of small merozoites which are the forms that
enter leucocytes. These forms, gamonts, are elongated rectangular bodies measuring 8-
12µm by 3-6µm. They are surrounded by a delicate capsule and stain pale blue with a dark
reddish-purple nucleus and have a numerous pinkish granule in the cytoplasm.

Life Cycle: Dogs are infected by ingestion of infected vector tick, Rhipicephalous
sanguineus, which contains sporocysts in its body cavity. Liberated sporozoites penetrate the
wall of the intestine of the dog, pass via the blood stream to the spleen, liver and bone
marrow, and here the enter tissue cells and become schizonts. Several generations of
schizonts occur, but ultimately merozoites enter the circulating leucocytes and become
gametocytes. Then the gamonts are ingested by a tick, in which they become associated in
pairs and the microgamont produces two non-flagellated microgametes, one of which
fertilizes the macrogamete to produce a zygote. This is motile (ookinete) and it penetrates the
intestinal wall to enter the haemocele of the tick, where it grows to become the oocyst.
Sporoblasts and then sporocysts are formed, each of which produces about 16 sporozoites.
On ingestion of the tick, these oocysts and sprocysts release sporozoites.

Pathogenesis and Symptoms: Parasites have been shown to be found in the myocardium of
domestic cats. Schizonts have an affinity to peripheral blood, spleen and lymph nodes. Still
inflammatory reactions from schizonts are scarce. The organism maybe found in apparently
healthy dogs, but it is associated with pathogenic effects only in the Far East and Africa.
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June 10, 2020 41
Clinical sign consists of an irregular fever, anaemia, progressive emaciation with the
enlargement of spleen. Lumbar paralysis may also be seen. Death occurs four to eight weeks
after the onset of clinical signs.

Diagnosis: Hepatozoon canis infection is diagnosed by blood smear demonstrating

gametocytes, or schizonts in spleen and bone marrow.

There is no treatment, and control is based on tick control.

SUBORDER - HAEMOSPORINA
FAMILY 10 - PLASMODIIDAE
Genus- Plasmodium
Species- gallinaceum
Geographical distribution- Commonly prevalence in tropical and subtropical regions.
Location and Host- It is primarily a parasite of domestic fowl in India and Nepal, but
experimentally other birds are also infected. In fowl Pre-erythrocytic phase is found in
macrophages of skin, Erythrocytic phage is found in RBCs and Exoerythrocytic phages occur in the
endothelial cells and the reticuloendothelial cells of the spleen, brain and liver. Sexual reproduction is
found in Culex, Aedes and Armigeses mosquitoes.
Morphology- The size of the oocyst is about 50-60µm in diameter and the size of sporozoite
is about 15µm in length. Schizonts are round to irregular in size and produce 8-30
merozoites.

Life cycle- The life cycle of P. gallinaceum is completed in two hosts. Asexual reproduction
is found in birds and sexual reproduction in Culicine mosquitoes. Birds are infected after
biting of the infected Culicine mosquito. The pre-erythrocytic schizont phase is found in the
macrophages and fibroblasts of the skin near the point of entry of the sporozoites. The
production of merozoites after 1 st pre-erythrocyte stage is called cryptozoites. The first pre-
erythrocytes (merozoites or cryptozoites) infect mostly macrophages and produce 2 nd stage of
cryptozoies called metacryptozoites. The metacryptozoites either enter into erythrocytes or in
the haemopoietic cells or endothelial cells. The merozoites of exoerythrocytic schizogony
also again start erythrocytic schizogony. The merozoites of erythrocytic schizogony from
RBCs are liberated and some merozoites of erythrocytic schizogony produce mega and micro
gametocytes. After sucking the infected blood, in Culicine mosquito (Aedes and Armigeses
etc), the male gametocyte undergoes further development into exflagellated form (6-8
flagella) and female gametocyte also develops.
After separation from the exflagellated form male gamete having one flagellum penetrates the
female gamete and forms a motile zygote called as ookinete. This ookinete penetrates the
midgut mucosa and comes to lie on the outer surface of the stomach. The nucleus of the
oocyst divides repeatedly to produce a large number of sporozoites. These are long with a
central nucleus and proceed to form oocysts. After 10-20 days of maturation, the oocyst
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ruptures and liberates the sporozoites into the coelom of mosquito. Some sporozoites enter
into salivary glands. These sporozoites infect the new host when the mosquito takes a blood
meal.

Pathogenesis and Symptoms- Chickens are particularly susceptible and even adult birds
may suffer; mortality may reach up to 80%. Birds become progressively emaciated and the
disease progresses. There is anaemia and spleen and liver enlargement is seen. Paralysis may
occur due to massive number of exoerythrocytic forms in the endothelial cells of the brain
capillaries.
Treatment-
Chloroquine 5 mg/kg b. wt.
Paludrine at 7.5 mg/kg. b. wt. and
Pyrimethamine at 0.3mg/kg b. wt. are effective against P. gallinaceum.

PHYLUM-APICOMPLEXA
CLASS- SPOROZOA
SUBCLASS- PIROPLASMIA
ORDER- PIROPLASMIDA
Family11 - Babasiidae
Genus- Babesia
a) Babesia bigemina

Geographical Distribution- This parasite is found worldwide in distribution but mostly found
in tropical and subtropical region of the world.
Location and Host- Asexual reproduction of B. bigemina is completed in red blood cells of
zebu and water buffalo and other cattle and horses. The sexual reproduction of these parasites is
completed in the gut wall of Boophilus annulatus, B. microplus and other species of ticks.

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Morphology- Babesia bigemina is a large piroplasma, 4-5µm in length and 2µm wide with a
rounded structure of 2-3µm in diameter. The organisms are found in RBCs and it has round,
oval or irregular forms, depending on the stage of the development of the parasite in RBC.

Life cycle of B. bigemina G=gamete, M=merozoite, Sg=salivary gland, Sk=strahlenkorper, Sz=schizont, T=trophozoite
To=transovarian, Ts=transstadial transmission, Z=zygote
Life Cycle- The life cycle of B. bigemina is completed in two hosts. Asexual life cycle is
completed in different species of ticks. This parasite causes the commonly lethal
Hemoglobinuria fever, Red-water fever, or Texas fever in cattle. Infection of cattle is initated with
the introduction of sporozoites into the blood through the bite of the infected tick Boophilus
annultus, B. microplus or other species of tick. Within the erythrocytes the parasites increase in
size and usually multiply by binary fission. The schizogony phase is rarely seen and probably
only occurs under abnormal conditions. Generally, multiplication is by binary fission and
budding processes represents asexual reproductive phase in this species. After dividing the
daughter cells may escape from the blood cells and reinvade others cells. Usually transovarian
transmission is seen in tick host. After the tick has taken a blood meal from the infected cattle,
intracellular parasites are released within its gut. Some of these individuals transform into
motile, vermiform isogametes. Ookinetes result from fertilization and penetrates the gut wall and
reaches into the ovary of the tick where it rounds up and transform into sporonts. These sporonts
secrete individual cyst walls within which each sporont divides to form naked sporoblasts. The
sporoblasts give rise to multinucleated sporokinetes as the result of rapid nuclear division. A second
cycle of development starts after involvement of the entry of vermicule cells into the malpighian
tubules and macrophages where they undergo further multiple fission.

Pathogenicity and Symptoms- The disease caused by B. biegemina resembles haemolytic

anaemia. High fever lasts from two to seven days or more and during this period a profound
anaemia frequently develops. There is haemoglobinuria and cardiac palpitation also occurs.
Initially, there is profuse diarrhoea and the mortality may be very high in acute cases, death
occurring four to eight days after the onset of clinical sighs. In the cerebral form of B.
bigemina, the parasites appear to accumulate and probably multiply in the cerebral
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June 10, 2020 44
capillaries. At postmortem in the cerebral form there is perivascular; perineuronal and
interstitial oedima observed throughout the brain and spinal cord.

Diagnosis- Diagnosis is based on the clinical signs and confirmed by the detection of
parasites in the peripheral blood.

Treatment-
 Pirevan or quirorium sulphate (Acaprin babesian), piroparv, acaprin, piroplasmin)=
1ml of 5% subcutaneous for per 50kg b.wt.
 Berenil or 4, 4 diamidinodiazoaminobenzene aceturate (Diminozene) 2-3.5mg/kg by
deep intramuscular injection for all babesiasis.
 Diampron (amicarbilide)-10mg/kg intramuscularly or subcutaneously for B. bigemina
and B. divergena
 Phenamidine 12mg/kg s/c ina 40% aqueous solution Imidocarb carbanilide.
 Imidazole 0.5-1mg/kg b.wt s/c.

Prevention- Prevention is by controling the tick population.

Family12 - Theileriidae
a) Genus- Thileria parva

Location and Host- This parasite is found in RBCs, lymphocytes and Histiocytes of cattles
in East, Central and South Africa and India. The vector for this parasite is Rhipicephalus
appendiculatus, but other species of Rhipicephalus and Hyalomma also transmit it.

Morphology- Two forms of this parasite are found in cattle. The forms which are found in
RBCs are mainly rod-shaped, 1.5- 2µm by 0.5-1µm in diameter. Sometimes round, oval,
comma and ring shaped forms may also occur. In the forms which are found in RBC, there is
no evidence of multiplication. The actively multiplying forms of this parasite occurs chiefly
in the cytoplasm of lymphocytes and occasionally, in the endothelial cells, especially of the
lymphatic glands and the spleen. These are called schizonts or Koch’s blue bodies. The shape
of schizont, which is an actively multiplying form, is circular or irregular shaped and
measures about 2µm to 12µm in diameter. Two forms of schizonts are recognized. The
schizonts which contain large chromatin granules 0.4-2µm in diameter are called
macroschizonts and produces macromerozoites. The other forms contain smaller chromatin
granules, 0.3- 0.8µm in diameter and are called microschizonts and produce micromerozoites.
Later they invade the RBCs and may represent sexual stages of the parasite.

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Life cycle- In Cattle
When the cattle are infected by infected tick vector then a large number of sporozoites are
released in blood. The transmission to the animal does not occur immediately on
attachment but sporozoites develop in the salivary gland during the first two to four days of
engorgement of nymph or adult stages of the tick. After the infection of sporozoites, the first
visible stages occur in the lymph nodes 5-8 days after infection. The first visible stages
are small rounded bodies with round nucleus. Schizogony of the parasite occurs in the
lymphoid and reticulo-endothelial tissues.They appear as multinucleated
macroschizonts, which is called as Koch’s blue bodies. Macroschizonts contain an
average of eight nuclei and the release of macro-merozoites from macroschizonts is not
observed. Later macroschizonts contain 50-120 nuclei. After the formation of macroschizont,
the formation of microschizont starts. The size of microschizont is smaller than
macroschizont. Micromerozoites are released and they penetrate the RBC’s of cattle. Later
on both forms form micro and macrogamont inside RBC.
In Tick Host- After the ingestion of intra erythrocytic stages of parasites by the tick host
(Boophilus and Rhipicephalus spp.), lysis of erythrocytes occurs and merozoites are liberated
which differentiate into sexual stages. In the lumen of the gut of infected nymphs, spindle-
shaped microgamonts develop from ring forms. These break up into several thread-like
microgametes after nuclear division and they develop thread like cytoplasmic
projections. Ring forms also develop into round forms, which are considered to be
macrogametes. Later both gametes fuse to form zygote. Six days afterwards, zygote appears
in the epithelial cells of the gut. There is an increase in size and progressively denser
cytoplasm occurs up to the third day after moulting to adult ticks. By the fifth day after
moulting a club shaped, motile ookinete is produced. The ookinete is passed into the
salivary gland which are found only in the granular secretory cells and round up into
sporoblast. Inside the sporoblast cell sporonts are formed by the processes of sporogony.
These sporozoites are formed after the fourth day of engorgement by the tick and ultimately
infect the new cattle host.

Pathogenicity and Symptoms – The disease caused by this parasite is called East Coast
fever. It is a serious disease with high mortality in susceptible stock, being characterized by
lymphoid hyperplasia, followed by exhausting of the lymphoid tissues and leucopenia. Zebu
cattle (Bos indicus) have a high level of natural resistance. The incubation period following
exposure is 10-25 days and the acute form of this disease is the most common, lasting 10-23
days. The main symptoms are high fever, swelling of lymphnodes, nasal discharge, swelling
of the eyelids and ears and diarrhoea with blood and mucus in the faeces. In adult animals the
mortality may be 95%. There is no haemoglobinuria because red blood cells are not
destroyed in case of T. parva.

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June 10, 2020 46
Diagnosis- The most satisfactory diagnosis is made by the demonstration of the schizonts in
material obtained from superficial lymphnodes or by spleen puncture. The forms in the
erythrocytes may be difficult to see.

Treatment:
 Oxytetracycline and Chlortetracycline- arrest macro and micro-schizonts
 Manoctone halts macroschizont production when injected on the first day of
fever  Oxytetracycline and chlortetracycline- 5-10mg/kg b.wt. I/M.
 Parvaquone – 20mg/kg b.wt. single dose I/M, or 10mg/kgb.wt. given 48hrs.apart.

b) Theileria annulata

Geographical Distribution- Parasite is mostly prevalent in North America, Middle and Far
East, Russia and Southern Europe, Iran, Nepal, India, Japan, Korea and Siberia. It is a highly
fatal disease of cattle in North America and trasmitted by Hyalomma species of tick.

Location and Host- Parasites are mostly found in the RBCs and lymphocytes of the spleen
and lymph node of cattle and buffalo.

Morphology- The parasite in the red blood cells are mostly round, oval or ring shaped and it
measures about 0.5-1.5µm in diameter. Sometimes rod shaped or comma shaped Anaplasma
like structure may also be found. The erythrocytic forms undergo binary fission with
formation of two daughter individuals which is the method of multiplication in T. annulata.
Macroschizonts and microschizonts are found in the lymphocytes of spleen and lymph nodes,
being similar to those of T. parva. Theileria annulata is easily transmissible by blood passage
and schizonts are found numerously in the circulating blood.

Life Cycle- The tick vectors for this parasite are various species of the genus Hyalomma. H.
savignyi (Syn. H. marginal) acts as the vector of T. annulata in India and Middle East. H.
dromedaril in central Asia.

The development of life cycle in the vertebrate host is related to that of T. parva and in T.
annulata intra-erythrocytic division is the most important method of multiplication but the
organisms also multiply in lymphoid cells.

There are two forms of erythrocytic stages which represent the gamonts. After ingestion of
the infected RBC by the tick, gamonts in the RBCs produce male and female gametes. In the
gut of the ticks male and female gametes fuse and zygote is formed. The size of zygote
increases and reaches about 10µm at 12 days of infection. At 13th day after division of the
nucleus a club shaped form is produced, which is called ookinete. These ookinete pass in the
acinar cells of the salivary glands (types II & III) of tick where spozoites are formed by the
process of sporogony. Later on sporozoites are released and mixed into saliva. The new hosts
are infected during engorgement by the infected tick.

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Pathogenesis and Symptoms- More pathogenic strains of this parasite occur in India, Iran
Russia and Israel. Mortality rate reaches up to 90% in some cases. The acute form of disease
occurs in all breed and all ages of cattle as well as buffalo and zebu. The incubation period is
9-25 days and the acute form of the disease may last from 3-20 days. The symptoms are:
 Marked rise of body temperature reaching40-410C depression
 Lacrimation
 A prograssive chronic anaemia is common
 Nasal discharge and swelling of the superficial lymphnodes
 Emaciation is rapid
 Haemoglobinuria may occur
The postmortem finding shows marked enlargement of spleen and liver. The lungs are
usually oedematous and lymphnodes may be swollen specially in the acute form of disease.

Diagnosis – It is based on the demonstration of parasites in the red blood cells or in smear of
material obtained from lymphnodes or spleen. Differentiation between T. annulata and T.
parva is not easy and diagnosis is based on the evaluation of the enzootic parasitic conditions
in the area.

Treatment-
Oxytetracycline and chlortetracycline- 5-10mg/kg b.wt. I/M.
Parvaquone– 20mg/kg b.wt. single dose I/M, or 10mg/kgb.wt. given 48 hrs. apart.

c) Theileria lawrenci
 Parasites are distributed in East and Central Africa and Angola.
 Parasite is mostly found in the RBCs of cattle and water buffalo and vectors are ticks
such as Rhiphicephalus appendiculatus and possibly R. duttoni.
 This parasite produces severe and fatal disease in cattle and water buffalo. The disease
caused by this parasite is called "Corridor disease". Corridor disease is highly
pathogenic and mortality reaches upto 80%.

d) Theileria mutans
 This parasite is mostly found in Africa, Asia, Australia and Russia.
 Parasite is mostly found in the RBCs of cattle and different species of ticks act as
vector some of which are Rh. appendiculatus, Rh. evertsi, H. bispinosa, H. punctata, B.
annulatus and B. microplus.
 Life cycle is same as T. parva.
 High fever and subclinical anaemia may also occur.

Theileria of sheep and goat-

Theileria ovis:
 Parasite is mostly endemic Africa, Asia, India, Russia and some parts of the Europe.
 Parasite is mostly found in the RBCs of infected sheep and goat. Different ticks Rh.
bursa, Rh. evertsi, D. sylvarum, H. sulcata etc. act as the vector of this parasite.
 Lifecycle is similar to T. parva
 The pathogenically is mild and there is seldom mortality occur.

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June 10, 2020 48
PHYLUM – CILIOPHORA
CLASS- KINETOFRAGMINOPHOREA
ORDER-TRICHOSTOMATIDAE

Family 13 - Balantidiidae
Genus- Balantidium coli
Geographical Distribution- Worldwide in distribution.

Location and Host- Parasites are mostly found in the large intestine of pig, man and
monkey.

Morphology- In case of B. coli, there are two morphological structures.

Vegetative form- It is a large form and average size of parasite is 50-60µm in length, but
sometimes reaches up to 150µm in length and it is easily observed in the stool of infected
animals. Body structure is oval in shape and is covered by cilia. There are two types of
nuclei, the micro-nucleus and mega-nucleus. The mega-nucleus is kidney shaped and
micronucleus is small rounded shaped. Food vacuoles and contractile vacuoles are present
in the endoplasm of the body.
Cyst- Cysts are ovoid to spherical shape and measure about 40-60µm in diameter. They are
faintly yellowish green in colour; the organism can be recognized within the cyst by the
macronucleus.

Reproduction- This parasite multiplies by transverse binary fission but conjugation also
takes place. Transmission to other host is by the cyst.

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June 10, 2020 49
Pathogenesis and Symptoms- Pigs acts as a primary host; and in pig B. coli is generally
regarded as a commensal. Under normal condition it is found in the lumen of the large
intestime and is associated with no change in the mucosa. But in heavy infection it may
invade the mucosa and cause superficial and even deep ulcerations. This is associated with a
mild to severe enteritis. In acute and at the time of fatal infection in pig it causes
haemorrhagic dysentery. This parasite is of zoonotic importance in Humans because usually
humans are infected by pig.

Diagnosis- By finding the cyst or some time also finding of trophozoite of the parasite in
faecal examination.

Treatment- Acute infections may be treated with the tetracycline antibiotics.

 Oxytetracycine –0.5gm orally
 Carbarsone dose of 250mg is given daily for ten days.
 Diodoquin- 300mg once daily for 5days.

ORDER: RICKETTSIALES (RICKETTSIA)

Genus: Anaplasma marginale

Geographical Distribution- Parasite is widely distributed throughout the tropical,

subtropical and some temperate areas of the world.

Location and Host- Cattle are the major hosts but infections also occur in the RBC of water
buffalo, bison, zebra, camel, sheep, goat and deer. The vector of this parasite is mainly
different species of ticks but stable flies, tabanids and mosquitoes also transmit the disease.

Morphology – Anaplasma appears as small, spherical body. Sometimes filamentous structure

may be found and the colour of the parasite is dark red in colour when stained inside the
RBC. The size of parasite is 0.2-0.5µm in diameter. Two morphological types of A.
marginale are found; a normal rounded form and a filamentous forms, both of which occur in
the majority of infected animals, but in some cases only the round form is present.

Life Cycle- Some 20 species of ticks and blood sucking flies such as tabanids, deer flies,
stable flies and mosquitoes transmit this parasite. Carrier cattle play an important role in the
epidemiology of infection although, deers have also been demonstrated to serve as carriers
for cattle and deer to deer transmission may occur in absence of cattle. Mechanical
transmission of Anaplasmosis is well known, and major and minor operations in cattle
husbandry such as dehorning, castration, vaccination, blood sampling may be responsible for
the transmission of Anaplasmosis.

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June 10, 2020 50
Pathogenesis and Symptoms – Anaplasmosis is essentially a disease of adult cattle and in
general, severe clinical infections do not occur until an animal is about 18monts of age. In
mature cows the incubation period is 15-36days with an average of 26days. The main
pathogenicity is high fever, anorexia develops and animals show severe anaemia. Mortality
rate may reach up to 80%.

Treatment- Tetracyclines (Chlortetracycline, Tetracycline and Oxytetracycline) – 6-10mg/kg

b.wt. I/M.

Anaplasma centrale
This organism is morphologically similar to A. marginale. It is names so because it is placed
in the centre of RBC. The infection is milder in comparison to that of A. marginale.

Anaplasma ovis
Parasite is mostly found in the RBCs of sheep and goat, it is usually non-pathogenic.

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June 10, 2020 51