By Dr. ASucharita, M.V.Sc., Veterinary Assistant Surgeon, Animal Disease Diagnostic Laboratory Kakinada..

 For confirmation of diseases.  For prognosis  To observe the effect of treatment and give directions for future therapy.  Improper collection & preservation may give False results. i.e : 1.Failure of Disease management programme. 2.Survey data on disease morbidity. and mortality becomes invaluble.

 Bacteriological  Viral  Parasitological  Histopathological  Biochemical  Clinical pathological  Toxicological

 1. Species, age and sex of animal  2. History  3. Date of collection of samples  4. Type of examination desired  5. Kind of samples and preservative used  6. Disease suspected  7. Number of animals affected and number of animals died  8.Symptoms  9. Autopsy findings

 1.Correct sampling  2. Appropriate preservation  3. Precise labelling and identification  4. Covering letter  5. Careful packing

1.Blood sears : for identification of bacteria and blood parasites.  Glass sides-clean & greese free.  Marking  Two thick & two thin sears needed. 2.Whole blood: for Bacteriological or virological examinations Collected in sterilised vials.

3.Citrated Blood: Useful in pathological examinations. Upto 6 hours – No need to add preservative Later – on ice. 4. Serum : Only potion in absence of symptoms Collect in test tubes and allowed to clot. After 6-12 hrs of refrigeration serum will be seperated. Urine : For Microbiological – 3 TO 5 ml For pathological (sugar,albumin,ketone bodies)-10ml

5.Faeces : Collect in sterile pencillin vials For Bacteria – on ice For parasitologiacl-10% formal Saline (1 in 3 parts) 6. Impression sears : taken from tissues for identification of bacteria & inclusion bodies. 7. Skin scrappings: for identification of fungal & Mite infctions. 8.Buccal epethelium : In case of FMD mucus membrane of vesicles in 50% Glycerine phosphate buffer. 9.Tissues : For Histopatological examinations 1sq cm piece Preservative : 10% formal saline.

Heart Blood : For Bacterial isolation Hot spatula is touched on the heart and sterile pastuer pipette is inserted.

 If not properly packed -preservative leak out  Packing Material : Packing Boxes: light unbreakable wooden boxes,thick paper cartons of aspirate sizes for keeping slides/vials/Bottles. Polythene bags : thick polythene bags for keeping the stomach contents & morbid tissues/faeces etc. Stuffing Material : For transportation of glass Ware absorbent cotton,husk,saw dust,paper pieces.

S.No.DiseaseMaterial to be collectedPreservativeRemarks 1. Foot & Mouth Disease a) Vesicular fluid from oral lesions for virus isolation or antigen detection b) Epithelial lining/buccal mucosa from lesions for virus isolation or antigen detection Neat in sterile vial on ice or in 50% Glycerine saline solution Can be sent by post or through a special messenger 2. Blue tongue a) Heparinised blood preferably at height of temperature for virus isolation or antigen detection b) Serum samples from maximum possible number of animals for serological tests c) Spleen from dead animals after postmortem for virus isolation or antigen detection Blood collected in Heparin Merthiolate Neat in sterile container on ice Through special messenger if it is neat material on ice and serum samples by post

S.No. Disease Material to be collected PreservativeRemarks 3.PPRa) Citrated or heparinised whole blood collected at the height of temperature b) Spleen and mesenteric lymphnode c) Swabs from lachrymal, nasal secretions d) Swabs from oral lesions On iceCan be sent by special messenger

S.No.Disease Material to be collected Preservative Remarks 4.Sheep PoxSkin scabs dry in a sterile container or in 50% glycerinated phosphate buffer solution 50% Glycerine phosphate buffer Can be sent by post

S.No. DiseaseMaterial to be collectedPreservativeRemarks 1.AnthraxDo not conduct postmortem examination a)Blood smears from ear vein as early as possible or within 6 hours of death for organisms identification b)If period is beyond 6 hours after death cut a piece of ear tip Air dried blood smears If possible on ice in dry bottle Can be sent by post or through a messenger

S.No.Disease Material to be collected PreservativeRemarks 2.Haemorrhagic Septicaemia a)Whole blood taken at height of fever b)Throat swelling exudates c)Piece of lung and spleen d)Smears of blood exudates- 2 thick and 2thin smears for staining On ice Air dried Neat material sent by special messenger

S.No.DiseaseMaterial to be collectedPreservativeRemarks 3.Black Quartera)Unstained smears from lesion b)Affected muscle tissue 1” cube in formal saline c)Affected muscle tissue 1” cube packed in salt d)Affected muscle tissue strips dried in shade e)Peritoneal and liver impression smears f)Swabs from the lesion Can be sent by post or through a messenger

S.No.DiseaseMaterial to be collectedPreservativeRemarks 4.Enterotoxaemia Stomach and intestinal contents with a few drops of chloroform in a sterile vial preferably a loop of intestine with contents inside Neat material on ice Sent by special messenger 5.Leptospirosisa)Freshly voided urine, Heparinised blood collected at height of temperature, kidney & liver b)Serum samples for serological tests c) Smears of blood and urine sediment On iceUrine or blood should reach lab within one hour

S.No.DiseaseMaterial to be collectedPreservativeRemarks 6.Johne’diseasea)A pinch of rectal mucous membrane and faecal sample for smear preparation and mesenteric lymph node impression smear for staining b)In dead animal affected portion of intestine particulary at ileocaecal valve for Histopathological examination 10% formal saline HP material by post. Other materials by special messenger

S.No.DiseaseMaterial to be collectedPreservativeRemarks 7.Foot rot a)Pus swabs from foot lesion in a sterile test tube for culture b)Smears of pus from foot lesions for staining On ice Air dried Pus to be sent immediately for anaerobic cultivation 8.MastitisAbout 5-8 ml of milk from affected teats in a sterile bottle. Preferably collect milk before treatment and if treatment is already given indicate the drugs used. If possible on ice Can be sent by messenger

S.No. DiseaseMaterial to be collectedPreservativeRemarks 9.Infectious abortions due to Brucellosis, Vibriosis, Leptospirosis etc. DAM: Uterine discharges swab, cervical swab, vaginal discharges, serum samples collected athe time of abortion and after on week and 3 weeks after abortion. Aborted foetus: Heart blood in sterile Pasteur pipette, stomach contents 1-2 ml in sterile test tube, foetal membranes if fresh SIRE: Serum samples for serological tests, preputial washings and semen for antigen detection & isolation For serum samples add Merthiolate and other materials in a sterile container Serum samples by post and other materials by special messenger

S.No.DiseaseMaterial to be collectedPreservativeRemarks 1Babesiosisa)Thin blood smears at least 6 numbers at the height of temperature b) Spleen and lymph nodes in case of dead animals c) Ticks from the body of the affected animal Air Dried In 50% Glycerin saline solution in dry bottle Can be sent by post

S.No.Disease Material to be collected PreservativeRemarks 2TrypanosomiasisBlood smears (thick) lymph node smears and citrated blood Air dried smears Blood through messenger or blood smears by post 3AnaplasmosisThin blood smearsAir driedCan be sent by post 4Coccidiosisa)Faecal sample 10 gms b)Scrapings of small and large intestine mucosa 10 % formal saline Can be sent by post

S.No.Disease Material to be collected PreservativeRemarks 1212 Parasitic infestations :Amphistomiasis Fasciolasis, Cestodes & Nematodes Microfilaria Faecal materials in a clean container a)Citrated blood to make wet films b)Thick blood smears in duplicate On ice Air dried By messenger By post 3Fungal DiseasesSkin scrapings, hair and hooves In a clean vial Sent by post

Commonly collected specimens Blood - from cases of bacteraemia or septicemia ( ex. Salmonella, Pasteurella etc.) Pus - from suppurative conditions (ex. Staphylococcus, Streptococcus, Enterobacteria) Faeces - from cases of enteritis and gastrointestinal infections (ex. Enteric viruses, enterobacteria, parasites) Respiratory discharge/ exudates - from cases of respiratory conditions (ex. TB, Candida)

Urine – from cases of urinary diseases (ex. Escherichia & Staphylococcus) Serum – for detection of antibodies (ex. Brucella, FMD etc.) Cerebrospinal fluid – from cases of nervous diseases (ex. JE, Rabies, Listeria)

Skin scrappings – from cases of fungal infections and ectoparasites (ex. Ring worm, Mange) Milk – from cases of mastitis and other systematic infections (ex. Brucella, Staphylococcus, Mycobacterium) Vaginal swabs – from cases of genital tract infections (ex. Bacterial infections, Chlamydia)

 Samples for toxicological analysis should be sent in saturated salt solution  A full written report of the clinical and necropsy finding along with requisition for which poison/poisons examination is warranted  The laboratory should be warned if there is any legal action that may arise out of the case  Details of the medication should be provided  Each organ may be sent in separate container with clean label  The containers in which specimens are collected should be chemically pure

Clean wide mouth glass bottles Saturated salt solution Add preservative up to two - thirds Slash the organs Apply Vaseline to stopper & seal Labelling Sample of preservative Keep in card board box & seal Paste address label Covering letter, details of materials, PM report & police report Send through authorized messenger & obtain receipt If not forwarded – Should be preserved for 6 months & Discard with  the written consent of police.  If further investigation needed send it to VBRI through ADDL(Histopathology,Bacteriology etc,)

 Quantity of the sample for toxicological analysis:  1. Serum- 5ml  2. Whole blood – 10 ml  3. Urine- 50 ml  4. Liver- 500g  5. Kidney- 50 g  6. Spleen- 50 g  7. Stomach or rumen contents – 500g

General Instructions Collect the sample as early as possible after death of the animal. Collect the Tissues Directly in Fixative. Hard organs like Liver, Kidney etc should be collected along with capsule.

In an outbreak fresh carcass must be autopsied, and at least 4-5 specimens must be collected from each case. All specimens collected for bacteriological / virus isolation must be collected aseptically and despatched on ice through courier with instructions to change ice on the way

10% formalin for histopathological examination Parasitic diseases – blood smears and stools / faeces from animals. Put a drop of formalin in stools and fix smears with alcohol.

FMD – 50% Glycerin phosphate buffer For confirmation of other viral disease, the material should be sent in 50% Glycerin saline

For sero – diagnosis collect paired serum samples in sterile vials and add antibiotics. For toxicological examination, collect fodder and stomach contents (500 gms approx) in rectified spirit or saturated salt solution.