HEPATOZOONOSIS

Hepatozoonosis is tick borne protozoan disease of carnibores eg.dog,cat etc.it is caused by protozoal agent Hepatozoon spp.it is also reported from wild animals like hyena,jaguar etc.In dogs hepatozoon infection occur in two forms Hepatozoon canis and Hepatozoon americanum.

Transmission-Unlike other vector borne diseases this is also transmitted by tick.Hepatozoon canis is transmitted by Brown dog tick Rhipicephalus sanguineus and Hepatozoon americanum is transmitted by Gulf host tick Ambylomma maculatum.Infection occur when an infected tick is eaten by dog.this is not transmitted by biting of tick.

Clinical sign-Signs of disease include high fever,loss of weight,loss of appetite,anaemia,weakness,nasal discharge,poor body condition,weakness of the rear limbs.sometimes diarrhoea is also observed.As the disease progresses lameness,severe muscle pain and inability to raise the weight on its hind limb.
                                                    watch video

Diagnosis-Confirmatory diagnosis of H.canis is done by microscopic examination of blood of infected dog.The microscopic detection of H.canis Gamont in blood smear stained with Giemsa stain.the gamonts always found in neutrophils.they are capsule shaped stained ice or light blue colour with pink nucleus detected in cytoplasm of neutrophils.the number on neutrophis are also increase.

                                         

Fig Gamont of H.canis in the cytoplasm of neutrophil.

                                                                           

Fig H.canis in neutrophil of dog.

Treatment-H.canis can be treated by Tab Doxycycline 10 mg/kg body weight oral for 10-15 days or clindamycine 5-10 mg/kg body weight for 10 days oral along with supportive therapy like appetite stimulant(syp. Aptiquick 1tsf bid for 10 days),hematinics(Syp Heamup 1 tsf bid for 10 days),liver tonic(syp Liv 52 1 tsf for 10 days).

Microscopic examination of Hepatozoon canis in Canine(German shepherd)

ANAPLASMOSIS

Anaplasmosis is a tick borne protozoan disease caused by several species of  rickesttsial parasite.Anaplasma marginale and Anaplasma centrale are two most common pathogens of cattle and buffalo but A.marginale primarly causes disease in cattle in india.Cattle and buffalo are two most common suseptible animal among these cross breeds are mainly effected.Sheep and goats are much less commonly effected.Anaplasmosis is also called "yellow beg" or "Yellow fever" because can develop jaundice.

Host-it causes disease in all animals e.g. cattle,buffalo,sheep,goat,horse and dog but cattle and buffalo are most commonly effected animals.

Transmission-It is not a contagious disease.it is commonly transmitted by ticks includes hyalomma,Rhipicephalus,Ixodid etc.It can be transmitted mechanically when red blood cells infected with A.marginale are inoculated into healthy and susceptible cattle.it can occur through needles or any infected surgical instruments.it can also transmitted by bite of fleae or mosquito.

Clinical sign-Clinical sigms are depend upon age.Cattle of all age can become infected but severity of disease is age dependent with cattle less then a year of old showing no or very mild clinical sign of disease but it will become a carriers.Carrier animal have immunity against anaplasma.Cattle 1-3 year of age will show more severe clinical signs.clinical sign manifested by high fever,anorexia,rapid shallow breathing,pale mucus membrane and decrease milk production.cow with light skin with initially look pale around the eyes and muzzle,but latter this can change to a yellowish color because of jaundice.Jaundice is develop due to destruction of red blood cells and their content being release into the blood stream.there is rapid weight loss.

Diagnosis-it is very difficult to diagnosed the anaplasmosis with theileriosis by clinical sign and symptoms because clinical signs of both disease are almost similar.confirmatory diagnosis is done by examination of blood of sick animal and identification of organism in the red blood cells.A.marginale is looks like a pinkish round or disk shape ponit with a wide zone around it and present of the margin of red blood cells.A.centrale is similar to A.marginale but it is found anywhere in the red blood cells but mostly found on center of RBCs.

Fig. Anaplasma marginale present at the periphery of erythrocyte

anaplasma margiale

Fig. Anaplasma marginale present at the periphery of erythrocyte

anaplasma marginale

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Fig. Anaplasma marginale present at the periphery of erythrocyte

Treatment-Treatment of anaplasmosis is most effective if given in early stage of disease.A single injection of Oxytetracycline @ 10 mg/kg body weight is administered I/V two times in a day is very effected.supportive therapy must be given for early recovery.like Hematinics(Ferritas @ 10 ml I/M on alternate days,liver tonics,ruminotorics.

BABESIOSIS

Babesiosis is one of the most fatal tick bornae protozoan disease of cattle and buffalo caused by babesia spp.it causes significant mortility,morbidity and loss of production.it is most important tick borne disease of cattle and buffalo in india.it is also occur in humans.In humans disease is called Lime disease.this disease infects most animals like cattle,buffalo,sheep,goat,horse and dog but it is most occurs cattle and buffalo even in dogs.this disease is highly dangerous then other protozoan diseases because this causes destruction of red blood cells of animal and produce anemia and cause death of animal within 3 to 4 days.

Causes- Babesiosis is protozoan disease caused by babesia spp. Babesia bovis and Babesia bigemina are two most common species of babesia which causes disease in cattle and buffalo.Babesia canis and Babesia gibsoni are most common strain of babesia which causes disease in dog.Babesia gibsona is mostly effect the dogs.Babesia equi and Babesia cabali causes disease in horses. Babesiosis is transmitted by tick Rhipicephalus spp.
B.bigemina is transmitted by Boophilus(Rhipicephalus) microplus-one host tick.
B.canis is transmitted by Rhipicephalus sanguineus-three host tick.

Host-Babesiosis is mostly occurs in cattle,buffalo,dog and horses sometimes occurs in sheep and goat even in human.cattle,buffalo and dogs are mostly infected by babesia in india. cross breeds of cattle,buffalo and dogs are highly.indiginous breeds of these animals are highly resistant for the disease.

Symptoms-Babesiosis is highly fatal disease.Babesia cause severe distruction of red blood cells result liberation oh hemoglobin and this hemoglobin is passes through the unine thats why clolur of uribe changed to coffee colour severe distruction of RBC causes anaemia.in acute cases the common sign is Haemoglobinurea,animal passes coffee colour urine,urination is freqent,high rise of temperature,swelling of superficial lymphnodes,severe anaemia,loss of apetite,reduce milk production.
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Fig Hemoglobinuria in cattle due to Babesiosis

Fig Hemoglobinuria in cattle 

Fig Hemoglobinuria in cattle

Diagnosis-Babesiosis can be confused with other diseases like post parturient hemoglobinuria,theileria etc.therefore confirmatry diagnosis by microscopic examination is very essential.under microscope morphology of babesia is depend of types of species.deferent species has different morphology but all species of babesia always present in the erythrocytes(red blood cells.Babesia bigemina is a largest species of babesia.it is occurs in pairs in the infected cell.its looks like two pear shape organism attached with each other but mainly single form is observed in the RBCs.B.bovis is smaller then bigemina.it is small,oval organism looks like large form of theileria sometimes its create confusion to the theileria.B.canis is also occur in pairs but the angle between to pairs is larger then B.bigemina.B.gibsoni is small,round half ring like structure.it smallest babesia species.B.equi occurs in four pairs which are attech to each other and looks like maltes cross structure.B.cabali is also occur in pairs looks like B.bigemina but it is only occur in horses.

Fig 1 pear shaped piroplasm of Babesia bigemina

Fig 2 paired piroplasm of B.bigemina inside the erythrocyte

Fig 3 B.bigemina 

Babesia bigemina

                               

Babesia bigemina in buffalo

                                                       Babesiosis in dog

Introduction-canine babesiosis is a tick borne haemoprotozoan disease of dogs occuring in topical and subtopical area of India particularly where vector ticks are present in sufficient number.The disease is caused by two species of Babesia i.e. Babesia canis and Babesia gibsoni both are transmitted by  transmitted by Rhipicephalus sanguineus-three host tick.Babesia canis is large form whereas Babesia gibsoni is small form of babesia.

Fig-Babesia canis larger then the Babesia bigemina

Babesia canis

Symptoms-it is characterised by high fever, anorexia,lethargy,haemolytic anaemia sometimes icterus,vomition and loss of body condition.The clinical syndrome produced by B.canis and B.gibsoni is indistinguishable but B.canis infection is generally consideres to be more acute.

Note-Haemoglobinuria is not observe in case of canine babesiosis but it is markedly observed in bovine babesiosis because canine RBCs are larger then the cattle that'swhy B.gibsoni are not able to breakdown of dog's rbcs.but sometimes haemoglobinuria can be observed in savere case of B.canis because it is largest form dof babesiosis.

                   
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Treatment-For therapeutic management the effected dog is treated with Diminazine aceturate@5mg/kg body wt intramuscular at 48 hr interval or Doxycyline @150 mg bid for 15 days orally.

Supportive therapy must be given daily untill recovery.supportive therapy includes Fluid therapy(dextrose saline @ 40ml/kg bodt wt till oral consumtion began),inj. Imferon(iron dextron 50 mg/ml) 2ml I/M on alternate day for 3-4 days,Liver tonic syp. Liv 52 or inj belamyl @ 1 ml I/M on alternate day for 3-4 days and syp. Polybion 1-2 tsf BID orally for 1-2 weeks.

                                                           

Choice of drug for babesiosis

THEILERIOSIS

Theileriosis is the tickbornae haemoprotozoan disease of livestocks,domestic and wild animals caused by theileria spp.Among livestocks this disease is highly prevalent in cattle and buffalo.the most important species which are highly effecting the cattle and buffalo are Theileria annulata which causes disease in india and Theileria parva causes disease in african countries.In india disease is also called as bovine tropical theleriosis.

Causes-Theileriosis is not contageous disease.it is tick bornae disease transmitted by Ixodid tick of genus Hyalomma spp.In india tropical theileriosis is mainly transmitted three host tick Hyalomma anatolicum which transmit theileria annulata.

Host-Theileriosis occurs in both domestic and wid animals.this disese is highly prevalent in cattle and buffalo.Among cattle disease is mostly occurs in cross breeds of cattle like holstein fzogen and jersy.Indian breeds of cattle are resistent for this dosease like sahiwal,hariyana etc.prevalence of this disease in other animals is low like sheep,goat,horse,dog and cat.

Symptoms-Theileriosis is fatal disese of livestocks.the sign and symptoms of the disease is depend upon severity of the infection.if infection is moderate animal reveals mild fever about 103-104°F,mild anaemia,animal become lethargy,ruduced feed intake ans sometimes animal shows difficult breathing.lymphnodes swelling is usually absent in low infection.if animal is severly infected then animal will show high rise in body temperature usually 105-106°F,reduce feed intake,suddenly drop in milk production,swollen lymphnodes,dyspnea,mucous membrane become pale sometime animal show juglar pulsation and there mey be presence of haemorrhage on mucous membrane of gum and conjuctiva or corneal opacity.

corneal opacity

Diagnosis-Sign and symptoms or blood parameters are not important tools for diagnosis of disease.the most common and reliable method of diagnosis of disease is examination of blood picture of infected animal,procedure is started with collection of blood from infected animal before given any antibacterial therapy.collection of blood either from juglar and ear vein.it's better to collect the blood and make blood smear on the spot imidiately collection of blood without mixing of any anticoagulant.otherwise collect the blood and send to the lab.blood smear is properly stained and examined under microscope.under microcroscope you should examined both erythrocyte and lymphocyte.piroplasm of theileria is present inside the erythrocyte and schizont stage is present in lymphocyte(koach's blue body).there are many form of piroplasm like oval,comma,annular,ring and rod form.annular and oval form are most common.sometimes lymphnodes aspiration or biopsy is also done for detection of koch's blue bodies in the lymphocytes.

Figure 1 erythrocytes of cattle are highly infected with pleomorphic intra-erythrocytic    piroplasm of theileria spp.

 

Figure 2 erythrocyte of bovine infected with pleomorphic theileria organism.

Figure 3  blood smear of cattle,erythrocytes are infected with annular form of theileria. annular form of theileria looks like small babesia piroplasm but not present in pair.

Theileria annulata

Figure 4 piroplasm of theileria spp.

Figure 5 Blood picture of goat contain oval shape piroplasm of theileria ovis,which is usually non-pathogenic.

piroplasm of theileria

 

 

Theileria annulata (Annular form)

Figure 6 piroplasm of theileria in the red blood cells

Fig.life cycle of theileria annulata  

 Video- https://youtu.be/V2iEqSwricA

 Video- https://youtu.be/V2iEqSwricA

Treatment-Treatment of Theileria depends upon the level of infection present in blood.use of oxytetracycline @ 10mg/kg body weight if infection is low.oxytetracyclin is 40% effective on Theileria.In case of heavy infection use the buparvaquone @ 2.5mg/kg body weight intramuscular along with oxytetracycline.single dose of buparvaquone is sufficient for treatment but sometimes double dose is required.sometimes combination use the berenyl (dimenazine aceturate) along with oxytetracycline is also effective.use of antipyretics, liver tonics, ruminotorics and hematinics require for supportive treatment.

PREVENTION-prevention of theileriosis is somewhat difficult because one tick is also capable of transmitting disease.prevention of disease is starts from the control of tick infestation for this regular check the body of animal.ticks are mainly present on thigh, wellly back and the inner side of hinD limbs.removal of ticks is by hand is a difficult job for this give the injection of ivermectin @ .2mg/kg body weight along with antihistaminic.this is highly effective for ectoparasites.there are many drugs available on the marjet in the liquid form for removal of ticks eg.  deltamethrin, flumethrin and cypermethrin.use one of them.regular clean place where the animal is rear.use of lime powder on the corner of the animal house because the effs of the ticks present on the corners of walls. 
one vaccine is also available for immunization of animals against theileriosis eg. Raksha vac T @ 3ML subcutaneous at the age of 3 year this gives immunity for one to two years.                 

TRYPANOSOMES

PHYLUM-SARCOMASTIGOPHORA  
ORDER-KINETOPLASTIDA

FAMILY-TRYPANOSOMATIDAE
CLASS-ZOOMASTIGOPHORA

• Members are all parasitic. They are found in blood, plasma, lymph and tissue fluids of mammals and birds, hence called as haemoflagellates.and may also have Pseudopodia.
• They have a characteristic leaf- like body with a single thread like flagellum pass along the body and remain attached to the body by the undulating membrane. Disease caused  is known as trypanosomosis.
• The nucleus is usually vascular and reproduction is generally by longitudinal BINARY FISSION.The nutrition is HOLOZOIC.
• In man, trypanosomes cause African sleeping sickness and Chagas' disease. In domestic animals, this disease is known as surra which means rotten. They are transmitted cyclically and mechanically by hematophagous flies.
• It affects a wide range of hosts and commonly seen in tropical countries like India, South Africa, and the UAE.
   Video- https://youtu.be/4dz6cdIUdqs
                        

  

  Trypanosoma

  
  

  

  

  Trypanosoma in cattle

  
  

  

  Fig.Trypanosoma evensi in blood smear of cattle

  
  

  

  
  

  

  
  

  

  
  

  
  

  

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MORPHOLOGY

• Body is usually elongated flattened leaf like in shape somewhat rounded with a vesicular nucleus(having one nucleus) and  kinetoplast located posterior to blepheroplast.kinetoplast contain DNA.
• they have single flagellum attached to the body by undulating membrane.the flagellum is arise from the BLEPHEROPLAST and passes anteriorly.
• Axoneme arises from kinetosome of basal granule attached to the body by undulating membrane and is continuous as free flagellum.
• Movement may be active or sluggish.
• Trypanosomes vary in shape.eg round,elongated leaf like.

• short and  stout stumpy forms
• long and slender forms ,and
• intermediary forms. Such trypanosomes with varying shape are called polymorphic trypanosomes are polymorphic trypanosomes.
• Some may be uniform in size – monomorphic tryps.

TRANSMISSION OF TRYPANOSOMA

• Multiplication is by longitudinal binary fission. Division starts form kinetoplast followed by nucleus and cytoplasm.
• With a single exception of T.equiperdum of equines,transmission from one vertebrates to another is carried out by blood sucking flies(TABANUS) in which development stages occur (cyclical transmission).
• Some of the trypanosomes are transmitted mechanically in which the infective stages are alive for a few minutes and they have to be transferrec to a new host for successful transmission.e.g. Trypanosoma evansi by Tabanus sp.
• Transmission is either cyclical or mechanical.
• CYCLIC TRANSMISSION
• Cyclical development occurs in the vector and may be in anterior station (salivaria) or in posterior station (stercoraria).
• In the anterior station development, infection is transmitted by inoculation by arthropod vectors when they suck blood from the host. eg. Trypanosoma vivax,T.congolense.T.brucei are transmitted by tse tse flies(GLOSSINA spp.)
• In the posterior station develpoment ,the metacyclic trypomastigotes accumulated in the hind gut are passed in the faeces of the arthropods. Infection of vertebrate host occur by contamination of skin or skin wound. e.g. Trypanosoma cruzi transmitted by REDUVIID BUGS.A relative non pathogenic trypanosomes like T.theileri and T.melophagium are transmitted by TABANUS FLIES.
• Any trypanosomes can be transmitted mechanically without cyclical changes experimentally this can be done by syringe passage.
• In nature this is accomplished by blood sucking insects like Tabanus, Stomoxys, Hippobosca sp., etc. which feed several times on different animals before repletion

DEVELOPMENTAL STAGE

usually 4 types of developmental stages...

• Amastigote (Leishmania like bodies, without flagellum)
  1. The body is spherical.
  2. Flagellum is absent or represented by short fibrils or degenerated into a tiny fibril inside the body.
  3. Kinetoplast is present.
  4. This stage is found in vertebrates and arthropods.
• Promastigote (Leptomonad,with a short free flagellum)
  1. Body is elongated.
  2. Kinetoplast and axoneme are towards the anterior tip of the body with no undulating membrane.
  3. It is mostly seen in invertebrates or cultures.
• Epimastigote (Crithidial,small free flagellum and short undulating membrane)
  1. Kinetoplast with axoneme is anterior to nucleus.
  2. Undulating membrane is short.
  3. This stage is seen in vertebrates but principally a stage in arthropod.
• Trypomastigote (Trypanosomes,complete undulating membrane and free flagellum)
  1. The body is leaf like or blade like.
  2. This is the form seen normally in blood films of infected animals.
  3. It is blade like form, kinetoplast posterior to nucleus and near to posterior extremity.
  4. Undulating membrane is well developed.
  5. Free flagellum is often present.
  6. Found in vertebrate host and also in arthropods.
  7. It is the infective stage for the invertebrate host.

  PATHOGENESIS

• Mainly anemia (Hemolytic Anemia).
  1. This may be due to immunological mechanism resulting in haemolysis and erythrophagocytosis.
  2. Trypanosome antigen attaches to RBCs and increases erythrophagocytosis.
  3. Anaemia may result due to reduction in half - life of circulating cells.
  4. Due to immune mechanism - enhanced haemolysis occurs; a hemolytic factor produced resulting in direct hemolysis of RBC.
  5. Anemia is also associated with disorders of clotting like thrombocytopenia and disseminated intravascular clotting ( DIC).
  6.  Damages to blood forming organs by trypanosomes.
• Hypoglycemia
•  Trypanosome absorbs glucose in blood leading to increased production of lactic acid. This leads to less intake of O2 by RBCs resulting in asphyxia. and acidosis.
• Serum potassium level is increased due to destruction of blood cells. Calcium and phosphorous ratio is disturbed.
• The lysed  trypanosomes release endotoxin resulting in toxemia and death.
• Due to the anaemia, mucous membrane is pale.
• Some of the clinical signs are
  1. Petechial hemorrhages and emaciation.
  2. Enlargement of spleen, lymph gland and liver.
  3. Congestion of mucosa of intestine, stomach, kidney and  bone marrow
  4. Oedema of dependant parts is common. Animal will be unable to get up.
  5. Keratitis  and conjunctivitis.
  6. Nervous symptoms.
  7. CSF section of the brain will show perivascular cuffing,  infiltration meningitis and  encephalitis.

TRYPANOSOMA EVANSI

      It is the first trypanosome shown to be pathogenic to mammals identified by Griffith Evans a British Vet in India.



HOST

• Natural hosts-Camel, horse, donkey, mule, ox, goat, pig, dog, water buffalo, elephant , mongoose , deer and other wild animals like fox, hyena and tiger.
• Experimental hosts- Many laboratory animals including mouse , rat, rabbit, guinea pig and chicken.

LOCATION

• Blood and lymph. The disease is called trypanosomosis.
• Name of the disease is different in different countries- most widely used is surra.
• Classical disease entity in Indian sub-continent  and occurs in horses and is known as surra.
• It is a Hindi word meaning rotten (or) putrified.
• Disease in camel is called Gufar; murrina – in panama; dorrengadera in Veninzula.
• T.equinum now regarded as dyskinetoplastic strain of T.evensi causes mal de caderes in horses in South America.

 PREVALENCE -Common in Northern Africa, Asia, Northern and South America

  STRUCTURE 

• Mean length varies considerably in different hosts and geographic strains.
• Typically they are 15-34 µm long with a mean of 24 µm.
• Most ones are slender (or) intermediate in shape. But stumpy forms also occur.
• Sporadic strains without a kinetoplast( dyskinetoplastic)  and visible with light microscope may arise  occasionally and  spontaneously or post-treatment with certain dyes (e.g. Acriflavin) and  drugs such diminazene aceturate.

INFECTION IN CATTLE OR BUFFALOES

• These two animals are  the main reservoirs of infection to equines. Infection is sub- clinical, mild and inapparent.
• Occasional outbreaks of acute disease are reported from many places. This may be due to lowered resistance in carrier animals following debilitating intercurrent diseases like rinderpest (or) FMD ,etc., stress after FMD vaccination and over work in draught animals.
• Introduction of new strain of parasite into newer areas  may result in acute form of the disease.  It appears as epizootic of variable severity. This can be highly fatal.

Clinical signs are

• High fever 41⁰C
• Intense excitement alternative to those of severe depression (coma)
• Animals move aimlessly in circles frequently falling down,  show colic, grinding of teeth, eyes staring wide open , breathing hard  and noisy
• Goring against wall apparent blindness stamping of feet following groaning, micturition, profused salivation, twitching of muscles followed by partial loss of senses  and prostration
• Parasitaemia due to factors unknown becomes too high; blood smears are seen with large number of parasites which occlude cerebral capillaries.
• Death may occur in 18 hr to 3 days.
• Sudden death may be mistaken for poisoning or  snake bite or  anthrax.
• In per acute cases death occurs in 2-3 hours. The nervous form of the disease show symptoms as above.
• Sub acute and chronic
  1. The animals look dull, sleepy
  2. lacrimation of eye, progressive emaciation rapid pulse
  3. Intermittent fever, oedema of leg, diarrhoea  and death
  4. Corneal opacity – twitching of muscles below eye
  5. Sub normal temp

INFECTION IN DOGS

• Disease is acute (or) fatal.
• Death in 2-4 weeks.
• Oedema is marked.
• The classical signs are corneal opacity, oedema of larynx resulting in voice changes and similar to that occur in rabies.

CAUSE OF DEATH

To sum - up, death in surra is mainly due to

• High fever, toxaemia
• Anaemia – PCV is less than 25 and 30 and decreased haemoglobin
•  damage of bone narrow  due to trypano toxin
• Increase in erythrocyte sedimentation rate   
• Hypoglycemia –  reduction of blood glucose  level by 30%
•  Exhaustion of glycogen reserves
•  Failure of liver cells  to compensate the loss in glycogen reserve

 DIAGNOSIS - TRYPANOSOMA EVANSI


  

• Clinical diagnosis can be done with history and clinical signs as described.

Laboratory diagnosis

    

• Direct examination or wet film examination
  • It is a quick method of detecting the organism by studying their movement and relative size . Species of trypanosomes involved can be guessed but it is to be confirmed by staining
• Peripheral blood smear examination
  • Thick and thin blood smears at the height of temperature is more desirable.
  • Parasitaemia is common in equines and canines but not readily seen in cattle, buffalo & camel.
  • Smears are stained with any of the GIEMSA OR LEISHMAN stain.
• Lymph node biopsy smears
  • Inject sterile normal saline into lymph node preferably prescapular lymph node with the help of a tuberculin syringe, massage and then aspirate.
  • It is risky to the operator
• Buffy coat smear
  • The suspected blood is spun and the buffy coat is examined for the presence of trypanosomes
• Biological test or animal inoculation test
  • The suspected blood is injected intra peritoneally  into susceptible laboratory animals like white rat, white mice, guinea pigs, rabbits .etc.,
  • Cryptic (or) sub- clinical trypanosomes in blood will multiply in these animals and cause death of these laboratory animals- Mouse: 48-72 hours; guinea pigs- 7 days; rabbits-60 days.
• Culture RPMI-1640,CAM
• Indirect test or Non-specific test to measure alteration of serum proteins
  • Mercuric chloride test
    • Reliable for surra in camels only.
    • This is done by adding one drop of suspected serum into 1: 20,000 solution of mercuric chloride.
    • A white precipitate is formed. In infected camels , this test gives  positive results in  2-3 weeks post infection
  • Stilbamidine test
    • Useful in bovine surra.
    • 0.5 – 2.5 ml of freshly prepared 10 % stilbamidine solution is taken in agglutinating tubes and one drop of inactivated serum is added.
    • In positive cases, coagulation occurs on the surface,begins to settle down in half a minute and dissolves in 5-10 minutes.
  • Formal get test
    • Useful in camel surra.
    • Two to four drops of formalin (40%) is added to 1ml of suspected serum and allowed it to stand .
    • In positive cases, gel formation occurs in 2-3 hrs.
  • Nitric acid test
    • One ml of 1.7 % nitric acid test is added to one drop of suspected serum.
    • In positive cases, white floccules are seen.
• Examination of other fluids
  • Cerebrospinal fluid will be examined in nervous form for the presence of trypanosomes but it is not important in veterinary practice.
  • Aqueous humor will be examined in case of blindness and in corneal opacity.
• Serological test
  • Some of the serological tests standardised are indirect haemagglutination, gel Diffusion, enzyme linked immunosorbant assay, complement fixation test and flourescent antibody test.
• Molecular test polymerase chain reaction (PCR) and PCR- restricted fragment length polymorphism (PCR-RFLP) are available for diagnosis and to know strain variation.