Veterinary Laboratory Manual

9th edition Veterinary Laboratory Service

Edition 1st edition 2nd edition 3rd edition 4th edition 5th edition 6th edition 7th edition 8th edition 9th edition

Year 1945 1947 1950 1961 1974 1981 1985 1995 2006

Under the Title Instructions for the Collection and Despatch of Material for Pathological and Bacteriological Examination. Instructions for the Collection and Despatch of Material for Pathological and Bacteriological Examination. Instructions for the Collection and Despatch of Material for Laboratory Examination Collection and Despatch of Material for Laboratory Examination Collection and Despatch of Material for Laboratory Examination Collection of Material for Laboratory Examination Collection of Material for Laboratory Examination Laboratory Specimen Submission Manual Veterinary Laboratory Manual

Prepared by staff of Elizabeth Macarthur Agricultural Institute, Menangle and Regional Veterinary Laboratories, Orange and Wollongbar

ISBN 0 7305 6656 0

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INTRODUCTION
This 9th edition of the Manual is published for the first time on the Web. Its aim is to provide accessible, up-to-date information to laboratory clients. The general format of the previous edition, edited by Graeme Eamens, has been retained. Many laboratory staff have contributed to this revised Manual. Also, field veterinarians have provided suggestions, which have been incorporated. The Manual includes information about specimen collection and submission in general, as well as for specific diseases. For further information about tests for diseases of commercial livestock, please contact your nearest Regional Veterinary Laboratory or Customer Service at 1800 675 623.

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........ 14 STORAGE OF SPECIMENS PRIOR TO DESPATCH.................................................... 11 BACTERIOLOGY .................................................. 18 STORAGE AND DESPATCH OF SPECIMENS ............................................................................................................................... 18 BLOOD ................................................................................ 20 STORAGE AND DESPATCH OF SPECIMENS ...................................................................................................................................................................................................................................................................................................... 16 COLLECTION OF SPECIMENS ................................. 25 STANDARD CONTAINERS AND EQUIPMENT ................. 4 TRANSPORT OF SPECIMENS.............................. 21 TOXICOLOGY .. 15 DESPATCH OF SPECIMENS .................................... 31 ABORTION IN SHEEP AND GOATS ............................................................................................ 3 PACKING OF SPECIMENS......................................................................................................................................................................................................................................................................................................................................................................... 18 COLLECTION OF SPECIMENS ............................................................................ 15 GENETICS .............................. 1 SPECIMEN SUBMISSION FORM ................................................................................................................... 16 STORAGE OF SPECIMENS PRIOR TO DESPATCH....................................................................................................................................................................................................................................................................................................................................... 15 STORAGE OF SPECIMENS AND DESPATCH OF SPECIMENS ................ 28 SPECIMENS (BY DISEASE OR SYNDROME).................................................... 31 ABORTION (GENERAL)....................... 27 STORAGE AND DESPATCH OF SPECIMENS ............................................................................................................................................ 19 STANDARD CONTAINERS AND EQUIPMENT ............................................................. 9 SPECIMENS (BY DISCIPLINE)............................... 16 COLLECTION OF SPECIMENS FOR GROSS PATHOLOGY ............................................................................................................................................................. 8 CHECKLIST OF EQUIPMENT FOR CLINICAL AND NECROPSY EXAMINATIONS ......... 18 PARASITOLOGY ............................................................................................................................ 18 BLOOD FILMS............................. 7 SPECIAL REQUIREMENTS FOR TESTING OF STOCK FOR INTERSTATE MOVEMENT .........................................................................................................................TABLE OF CONTENTS INTRODUCTION .................................................................... 27 COLLECTION OF SPECIMENS .............................................................................................................................................................................................................................................................................................. 14 BIOCHEMISTRY ........................................................................................................................................................................................................... 21 TYPES OF SEROLOGICAL TESTS ...................................................... 24 STANDARD CONTAINERS AND EQUIPMENT .................... 18 STANDARD CONTAINERS AND EQUIPMENT ......................................................................................................................................................................................................................................................................................................................................................... 17 HAEMATOLOGY.. 20 COLLECTION OF SPECIMENS ... 1 LABORATORY CHARGES................. 31 ABORTION IN HORSES ............................................................................................................................................................... 19 STORAGE AND DESPATCH OF SPECIMENS ......................................................................................................................................................................................................................................... 31 DISEASES OF LIVESTOCK .....................................................iii VETERINARY LABORATORY DIRECTORY ................... 24 COLLECTION OF SPECIMENS ................................................................................................. 25 DIAGNOSIS OF VIRAL DISEASE ...................... 15 STANDARD CONTAINERS AND EQUIPMENT .............. 24 STORAGE OF SPECIMENS PRIOR TO DESPATCH............. 16 STANDARD CONTAINERS AND EQUIPMENT ............................................................................................................................................. 1 CONDITIONS FOR ACCEPTANCE .......................................................................................................................................................................................... 16 GROSS PATHOLOGY ............................................................................ 15 COLLECTION OF SPECIMENS .................................................................................. 14 COLLECTION OF SPECIMENS ............................................................... 1 COLLECTION AND LABELLING OF SPECIMENS .............................................................................................................................................................................................................. ix SPECIMENS (GENERAL) .......................... 31 ABORTION IN CATTLE.......................................................................................................................................................................... 11 COLLECTION OF SPECIMENS ............................................... 32 VETERINARY LABORATORY MANUAL IV ............................................................................... 11 STORAGE OF BACTERIOLOGICAL SPECIMENS PRIOR TO DESPATCH ................................................... 14 STANDARD CONTAINERS AND EQUIPMENT ............................................................................................................................................................................................................ 5 DISEASE SURVEILLANCE .......................................................................................................................................... 18 PREPARATION OF BLOOD FILMS ................................................................................................................................................................................................................................................................................................................................................................................................................ 24 VIROLOGY ................... 11 STANDARD CONTAINERS AND EQUIPMENT .............. 16 HISTOPATHOLOGY .................................................. 19 SEROLOGY.................................. 13 DESPATCH OF SPECIMENS ........................................................................................................................................... 6 SPECIAL REQUIREMENTS FOR TESTING OF STOCK FOR OVERSEAS EXPORT.............................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................. 16 STANDARD CONTAINERS AND EQUIPMENT .......................................................................................................................................................................................

......................................................... 45 CONTAGIOUS EQUINE METRITIS (CEM) ................................................................................................................ABORTION IN PIGS ........................................................................................................... 47 DERMATOPHILOSIS ............. 43 CLOVER DISEASE..................................................................................................................................................... 32 ACETONAEMIA................................................................................................................................................................................................................................... 42 CHLAMYDIAL INFECTIONS ............................................................................................................................................................................................................... 44 COLIBACILLOSIS................................... 46 CYANIDE POISONING................................................................ 33 ALPHA MANNOSIDOSIS OF CATTLE.... 47 CYSTICERCOSIS.......................................................................... 47 DRENCHING MORTALITIES ............................................................................................................................................................................................................. 33 ANAPLASMOSIS............................................... 32 ACTINOBACILLUS SEMINIS INFECTION IN RAMS ........................................................................................................................................................................................................................................................................................................................................... 43 CITRULLINAEMIA ........... 38 BLACK DISEASE.................................................................................................................................................................................................................................................................... 33 ANTHELMINTIC RESISTANCE........................................................................................................................................................................................................ 37 ASPERGILLOSIS ............................................. 38 BLOAT ....................................................................... 46 CRYPTOSPORIDIOSIS.................................................................................................................................... 48 ENCEPHALOMYELITIS AND ENCEPHALITIS ................................ 48 VETERINARY LABORATORY MANUAL V .......... 38 BALANO POSTHITIS IN RAMS......................................................................................................................................................... 40 BRACKEN FERN POISONING................................................................................................................................... 48 DWARFISM IN DEXTER CATTLE........................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................ 34 ANTHRAX....................................................................................... 41 CAMPYLOBACTERIOSIS OF PIGS................................................. 40 BRUCELLOSIS OVINE................................................................................... 41 CAMPYLOBACTER ENTERITIS ................................................................. 39 BOTULISM .......................................................................................................................................................................................................................................................................................... 33 ANAEMIA................. 45 COPPER DEFICIENCY ............................. 39 BOVINE LEUCOCYTE ADHESION DEFICIENCY (BLAD) ....................... 42 CARDIOMYOPATHY AND WOOLLY HAIRCOAT (CWH) SYNDROME ..................................................................................... 38 BABESIOSIS ........................................................................................................................................................................................................................................................... 32 AFLATOXICOSIS ............................................................................................................................................. 39 BORDER DISEASE IN SHEEP ..................................................................................................................................................................................................................................................................................................................................... 40 CALCULI.......... 41 CAMPYLOBACTERIOSIS OF CATTLE..................................................................................... 45 COPPER POISONING OF SHEEP AND CATTLE ........................................ 36 ARTHRITIS AND POLYARTHRITIS ................................................................................................................................................................................................................................ 38 'BIG KNEE' IN GOATS ................................................................................................................................................................. 37 ATAXIA ................ 45 CONTAGIOUS OPHTHALMIA........................................................................................................................................................................................................................................................................................... 39 BOVINE LEUKOSIS .. 32 ACTINOBACILLOSIS AND ACTINOMYCOSIS ................................... 43 COCCIDIOSIS ......................... 33 ANNUAL RYEGRASS TOXICOSIS (ART) ............ 40 BRUCELLOSIS PORCINE ................................................................................................................................ 37 ATROPHIC RHINITIS OF SWINE .. 44 CONGENITAL ABNORMALITIES................................. 33 AKABANE DISEASE IN SHEEP AND CATTLE....... 36 ARSENIC POISONING........................................................................................................................................................................................................................................................................................................................................................... 47 DEFICIENCY OF URIDINE MONOPHOSPHATE SYNTHETASE (DUMPS) ...................................................................................................................... 38 BLACKLEG .............................................................................................................. 46 CORYNEBACTERIUM EQUI IN HORSES OR PIGS ........... 38 BETA MANNOSIDOSIS OF GOATS .......................................... 39 BOVINE VIRUS DIARRHOEA .............................. 43 COBALT DEFICIENCY ...................................................................................................................................................................... 38 BLUE-GREEN ALGAL POISONING.............................................................................................................................................................................................................................................................................. 38 BENIGN FOOTROT............................................................................................................................................................................................................................................................................... 42 CASEOUS LYMPHADENITIS (CLA) ..................................................................................................................................................................................................................................................................................................................................................................... 41 CAMPYLOBACTER ABORTION OF SHEEP .................. 47 DIARRHOEA............................................................................................ 33 ALGAL POISONING .................................................................................................................................................................... 37 ARTHROGRYPOSIS AND HYDRANENCEPHALY.. 40 BRUCELLOSIS BOVINE ....................................................................................................................... 44 COMPLEX VETERBRAL MALFORMATION (CVM) ....... 39 BOVINE MALIGNANT CATARRH ..................................................................................................................................................... 42 CAPRINE ARTHRITIS ENCEPHALITIS (CAE).................................. 45 CONTAGIOUS PUSTULAR DERMATITIS .............................................................................................. 44 COCCIDIOSIS – HEPATIC IN RABBITS............................................................................................................................................................................................................

............... 70 MALNUTRITION ..................................................................................................................................................... 71 ALPHA-MANNOSIDOSIS ................................................................................................................................................................................................................................................................ 51 EXOTIC DISEASES.................................................................................................................................................................................................................... 62 INFERTILITY IN CATTLE...................................................................................................................................................................................................................... 56 FUNGAL INFECTIONS (OTHER THAN MYCOTOXICOSES).................... 59 HEPATOSIS DIETETICA........................................................................................................................................................... 50 EQUINE VIRAL ARTERITIS (EVA).................OVINE .................................................................................................................................................. 51 FACIAL ECZEMA ...................................................................................................................... 48 ENTEROTOXAEMIA ........................................... 60 HERPESVIRUS VULVOVAGINITIS/BALAN0POSTHITIS IN GOATS.............................................. 59 HELIOTROPE POISONING . 63 JAUNDICE ....................................................................................................................................................................................................................................................................................................................................................................... 63 ITCHMITE IN SHEEP .............................................................................................................................................................................. 53 FOOTROT IN SHEEP.............................................................................................................................................................................. 62 INHERITED DEFECTS............................................................................................................................. 59 HAEMOPHILIA .................................................................................................... 60 HYPOGLYCAEMIA........................................................................................................................................................................................................................ 66 LEPTOSPIROSIS .................................................................................................................................................... 53 FOOTROT IN GOATS AND CATTLE . 49 ENZOOTIC BOVINE LEUKOSIS (EBL)......... 50 EQUINE BABESIOSIS........................................................................................................................ 62 INHERITED CONGENITAL MYOCLONUS ..................... 51 EXUDATIVE EPIDERMITIS IN PIGS..... 62 IRON TOXICITY SYNDROME IN PIGLETS ........................................................................................................................................................................................................................................................................ 49 ENZOOTIC HAEMATURIA OF CATTLE .................................. 50 EPHEMERAL FEVER............................................................................................................................. 70 MANNOSIDOSIS ..... 66 LICE RESISTANCE TO INSECTICIDES IN SHEEP ............................................................................... 58 GRASS TETANY ......................................................................... 61 INFECTIOUS BOVINE RHINOTRACHEITIS (IBR) AND INFECTIOUS PUSTULAR VULVO VAGINITIS (IPV) ....................................... 61 ILL THRIFT ........................................................................................................................................................ PIGS ...................................................................................................................................................................................................................................................................................................................... 71 VETERINARY LABORATORY MANUAL VI ............................................ 49 EPERYTHROZOONOSIS OF SHEEP.................................................................................................ENCEPHALOMYOCARDITIS (EMC) OF PIGS ..................................................... 71 BETA-MANNOSIDOSIS .................................................................... 60 HYPOCALCAEMIA ........................................... 59 HENDRAVIRUS INFECTION.............................................................................................................................................................................................................................................................................................................. 48 ENZOOTIC ATAXIA............................................................................................................................................................................................................................................. 59 HAEMOGLOBINURIA............................................................................................................................................................................................................................................................................................................................................................................................................................ 52 FASCIOLIASIS IN SHEEP AND CATTLE .................................. 50 ERYSIPELAS.................................................................................................................................................... 57 GOITRE ........................................................................................... 52 FISTULOUS WITHERS ........................................................................................................... 61 HYPOPHOSPHATAEMIA.................................................................................... 52 FACTOR XI DEFICIENCY ................................................................................................................................... 63 KIKUYU POISONING ..... 49 ENZOOTIC PNEUMONIA OF PIGS ...................................... 68 LISTERIOSIS................................................................................................... SHEEP................. 61 HYPOMAGNESAEMIA ..................................................................................................................................................................................................................................... 66 LEAD POISONING ...................................................................................................................... 57 GENETIC DISEASES ............................................................................................................................ 63 JOHNE'S DISEASE .. 70 MANGE.......................................... 52 FACTOR VIII DEFICIENCY .......................................................................................................................................................................................................... 69 LYME DISEASE................................................................................................................................................................................................. 52 FOCAL SYMMETRICAL ENCEPHALOMALACIA (FSE) ............................................... 52 FLEECE ROT ............... 70 MALIGNANT OEDEMA ........................................................................................................................................................................................................................................................................... 50 EQUINE INFECTIOUS ANAEMIA (EIA) ........................................................................................................................................................................................................................................................................................................................................... 53 FOOT ABSCESS IN SHEEP ............................................................... 50 EPIDIDYMITIS .................... 59 HEEL ABSCESS ..... 69 LYSSAVIRUS ....................................................................................................................................................................................................................................................................................................................................................................................... 68 LUPINOSIS................................................................................................................................................. 58 HAEMAGGLUTINATING ENCEPHALOMYELITIS VIRUS (HEV) INFECTION OF PIGS................................................................................................................................................................................................................ 56 GENERALIZED GLYCOGENOSIS........................... 59 HAEMATURIA ................................................. 58 GRAIN POISONING ..................................... 68 LIVER FLUKE INFECTION....... 60 HISTOPHILUS OVIS/ HAEMOPHILUS SOMNUS INFECTIONS .....................................

........................................... 93 PYRROLIZIDINE ALKALOIDOSIS .................................................................................................................................................................................................................................................................................................................................................................................................... 76 NECROBACILLOSIS .............................................................................................................................. 91 PORCINE INTESTINAL HAEMORRHAGE SYNDROME............................. 89 PORCINE ENTEROVIRUS ENCEPHALOMYELITIS ................................................................................................ 80 PARVOVIRUS INFECTION IN PIGS ............................................................................................................ 79 OSTEOMALACIA.................................................................................................................................... 94 RINGWORM (DERMATOMYCOSIS) .......... 93 PSEUDOCOWPOX ......................................................................................................................................... 76 MYCOTOXICOSIS................................................................. 92 PORCINE STRESS SYNDROME (PSS) ............. 87 PHOTOSENSITIZATION ................................................ 93 Q FEVER ................................................................................................................................. 79 OXALATE POISONING .......................................................................................................................................................................................................................................................................................................................... 86 PHALARIS POISONING............ 91 PORCINE ENZOOTIC PNEUMONIA ....................................................................................................................................................................................................................................................................................................................................................................................................................................................................................... 79 PARAMPHISTOMIASIS.................................................................................... 91 PORCINE MYOCARDITIS (PMC) ........................................................................................................................................................ 79 OSTERTAGIOSIS............................................................... 88 POISONING (PLANT).................. 73 MASTITIS (OVINE)..................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................... 89 POLYARTHRITIS ............................................................................. 87 PHOSPHORUS DEFICIENCY........................................................................................... 93 PYOGENIC INFECTIONS ................................................................................................................................................................................................... 73 MENANGLE VIRUS INFECTION............................................................................................................................................... 84 PASTEURELLOSIS .................................... 88 POLIOENCEPHALOMALACIA (PEM) ................... OSTEOPOROSIS........................................................................................................................... 87 PLANT POISONING ................................ 93 PROTOPORPHYRIA ................................................................................................................... 94 RED GUT IN SHEEP .......................................................................................................................................................................................................................... 94 RHODOCOCCUS EQUI LYMPHADENITIS IN PIGS ...................... 89 PORCINE COLITIS.................... 76 NEOPLASMS.......................................................................................................................................MAPLE SYRUP URINE DISEASE (MSUD) ....... 74 MYCOPLASMOSIS ........................................... 94 RHODOCOCCUS EQUI PNEUMONIA IN HORSES ........... 88 IDENTIFICATION OF SUSPECT POISONOUS PLANTS .................................................................................................................................................................................................... 92 PORCINE PROLIFERATIVE ENTEROPATHY....................................................................................................................................... 73 MASTITIS METRITIS AGALACTIA SYNDROME IN SOWS.......................................................................................................................................... 89 POMPE'S DISEASE .............................................................................................................................................................................................................................................. 87 PIGLET ANAEMIA ......................................... 80 PARASITES (EXTERNAL) ..... 71 MASTITIS (BOVINE) ........................................................ 76 NASAL GRANULOMA OF CATTLE ..................................................................................................... 77 NITRATE-NITRITE POISONING .................................................................................................. 78 OEDEMA DISEASE OF PIGS .............................................................. 79 PAPULAR STOMATITIS OF CALVES.......................... 80 PARASITES (INTERNAL)............................................................................................ 91 PORCINE PLEUROPNEUMONIA ........................................................ 85 PEPSINOGEN ESTIMATIONS FROM SERUM OR PLASMA................................................................................................... 77 NEOSPOROSIS ..................................................................................................... 79 OVINE BRUCELLOSIS..................................................................................................................... 78 OSTEOCHONDROSIS IN PIGS ................................................................................................................................................................................. NUTRITIONAL ..................................................................... 94 VETERINARY LABORATORY MANUAL VII ...................................................................................................................................................................................... 85 PESTIVIRUS INFECTION ..................... 85 PERINATAL MORTALITIES IN SHEEP AND GOATS ............................ 87 POISONING (CHEMICAL)........................................................................................................................................................................... 73 METABOLIC DISEASES ....................................................................................................................................................................................................................................................... 93 PYELONEPHRITIS (BOVINE) .......................... 85 PERINATAL MORTALITIES IN CATTLE...................................................................................... 74 MUCOSAL DISEASE.............................................................................................................................................................................................................................................................................................................................................. 79 PARAKERATOSIS OF SWINE ....................................................................................................................................... 87 PINKEYE ........................................................................................................................................ 78 OPHTHALMIA................................................................................... 72 MASTITIS (CAPRINE) .................... 77 NERVOUS DISORDERS ....................................................................... 75 MYCOTIC DERMATITIS.............................................................................................................................................................................................................................................................................................................................. 87 PNEUMONIA .................. 74 MUSCULAR DEGENERATION............. 74 MULBERRY HEART DISEASE ....................................................................................................................................................... 78 ORGANOCHLORINE AND ORGANOPHOSPHATE POISONING.......................................................... 92 PREGNANCY TOXAEMIA........................................................................

........... 95 SALT POISONING IN PIGS ............................................. 99 TETANUS ................................................................................... CRUSTACEANS AND SHELLFISH ALREADY ENCOUNTERED IN NSW....................................................................................................... 98 SWAINSONA POISONING....................................................... 109 STANDARD CONTAINERS AND EQUIPMENT ............................................ 96 SELENIUM DEFICIENCY .......................................................................................................................................................................................................................................................................................................................................................................... 95 SCOURING......................................................................................................................................................... 101 ULCERATIVE SPIROCHAETOSIS IN PIGS.............................................................. 108 PULLORUM DISEASE ............................................................................................................................................. 95 SALMONELLOSIS...................................................... 111 HISTORY AND DIAGNOSIS........................................ 99 TICK FEVER OF CATTLE ...... 105 STORAGE AND DESPATCH OF SPECIMENS ................................................................................. 107 MAREK'S DISEASE.................................................................................................................................................................................................................................................................................... 108 DISEASES OF CAGE BIRDS.......................................................................................................................... 111 STANDARD CONTAINERS AND EQUIPMENT ................................................................................................................................................................................................................................................................ 108 TUBERCULOSIS (AVIAN) ..................................................................................................................................................... 106 BIG LIVER AND SPLEEN DISEASE ......................... 102 URINE EXAMINATION ................................................................ 100 TOXOPLASMOSIS IN SHEEP AND GOATS ................................................................................................................................................................................................................................................................................................................................................................................................................................................................ 94 ROTAVIRUS INFECTION........................................................................................................... 102 ULCERS................................................ 99 TEAT LESIONS ................................................................................................... 107 MYCOPLASMOSIS (AVIAN) ............................................................... 99 TOXAEMIC JAUNDICE ....................................................................... 114 WATER TESTING... 102 DISEASES OF POULTRY................................................................................................................................ 105 AVIAN ENCEPHALOMYELITIS (AE)...................................................... 96 STRYCHNINE POISONING .... 102 UREA POISONING............................................................................................................................ 100 TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY (TSE)........................................................................................................................................................................................................................................................................................................................................................................................... 105 COLLECTION OF SPECIMENS ....... 105 STANDARD CONTAINERS AND EQUIPMENT ..................................................................................................................................................................................... 112 SOME DISEASES OF FISH.... 95 SARCOSPORIDIOSIS.......................................................................................................................... STORAGE AND DESPATCH OF SPECIMENS........ 116 VETERINARY LABORATORY MANUAL VIII ........................ 98 TAPEWORM INFESTATION IN SHEEP ............................................................................................................................................................................ 108 RETICULOENDOTHELIOSIS (RE) ..................... 109 COLLECTION OF SPECIMENS ..................................................................................................................... 105 SEROLOGICAL TESTS AVAILABLE FOR POULTRY ..................................................................... 100 TOXOPLASMOSIS IN OTHER ANIMALS .......................................................................................... 107 PASTEURELLOSIS (AVIAN) ............................................................................................ 110 DISEASES OF FISH.......................................... GASTRIC IN PIGS ............................................................................................................................................................................................... 109 DISEASES OF BEES ............................................................................. 113 FEEDS AND PASTURE ANALYSIS ............................................................................................ 100 TOXOPLASMOSIS IN CATS ....................................................... 95 SCABBY MOUTH ................................................................................................................................................................................................................................................................... 106 LEUKOSIS ..............................................................................................................................................................................ROCK FERN POISONING ............................................................................. 101 TUBERCULOSIS ............................................................................ 111 COLLECTION...................................... 95 RYEGRASS STAGGERS .......................................................................... 109 STORAGE AND DESPATCH OF SPECIMENS ........................................................... 101 TRICHOMONIASIS OF CATTLE ........................................................................................................................................... AVIARY BIRDS AND RACING PIGEONS.................................................................................................................................... 96 SEMEN EXAMINATION ............................................................ 98 SWINE DYSENTERY ..............................................................................................................................................................................................

Camden 2570) Officer in Charge: Pathologists: Keith Walker Steven Hum Rod Reece Richard Luong Ph: 02 4640 6333 Fax: 02 4640 6300 Ph: 02 6391 3858 Fax: 02 6391 3899 Contacts Ph: 02 4640 6327 Fax: 02 4640 6400 Elizabeth Macarthur Agricultural Institute (Main Reception) Orange Regional Veterinary Laboratory Agricultural Research and Veterinary Centre Forest Road Orange NSW 2800 Graham Bailey Patrick Staples Erica Bunker Regional Veterinary Laboratory Wollongbar Agricultural Institute 1243 Bruxner Highway Wollongbar NSW 2477 Officer in Charge: Pathologists: Graeme Fraser John Boulton Paul Gill Officer in Charge: Pathologists: Wollongbar Ph: 02 6626 1103 Fax: 02 6626 1276 VETERINARY LABORATORY MANUAL IX .VETERINARY LABORATORY DIRECTORY Phone ‘Customer Service’ at 1800 675 623 to contact any of the following veterinary laboratories Location Menangle Address Regional Veterinary Laboratory Elizabeth Macarthur Agricultural Institute ‘Camden Park’ Woodbridge Road Menangle NSW 2568 Postal address: (PMB 8.

. cage birds. CONDITIONS FOR ACCEPTANCE The Veterinary Laboratories examine specimens from all livestock. which can be submitted by primary producers. The Genetics section. racing dogs and horses. This information will allow the laboratory veterinarian interpret the laboratory results and suggest additional testing as appropriate. in cases of suspected malicious poisoning). Laboratory charges are subsidised for testing in the following circumstances*: • Notifiable disease • Emergency (exotic) disease • National TSE Surveillance Program • Mortality investigations *See ‘Disease Surveillance’ below for more details. The source of the information will remain confidential unless otherwise required by law or regulatory policies. 1 . Details on the scope of this accreditation can be found on the NATA website at http://www. Specimens are generally only accepted on referral from a veterinarian. Where the submitter expects the work to be funded (entirely or partially) by NSW Department of Primary Industries. and are accredited by the National Association of Testing Authorities (NATA).SPECIMENS (GENERAL) All NSW Department of Primary Industries Veterinary Laboratories operate according to ISO 17025. Often they have special procedures to be followed with respect to sampling/examination.au/agriculture/vetm anual/submission/lab-charges#fees.gov.nsw. Laboratory fees for tests not listed can be obtained by calling Customer Service at 1800 675 623. INFORMATION ANALYSIS Test results and findings may be provided to other authorised staff and used for statistical. surveillance. The information assists disease and residue control programs and underpins marked access for agricultural products. specimens are unlikely to be admissible as evidence in criminal prosecutions and civil actions. The Pathologist will consult with the submitter before exceeding this amount. Exceptions include WormTest kits. many find it useful to indicate a maximum laboratory fee on the Specimen Advice Form.g. at EMAI also accepts bovine hair samples collected by cattle owners for genetic disease testing where certification is not required. it is policy to examine only specimens where the health of the flock.dpi. poultry. care and transport of exhibits. those bodies should be contacted by phone and consulted before any action is taken. custody. CASES LIKELY TO INVOLVE CRIMINAL INVESTIGATIONS Examination is not undertaken on specimens likely to involve criminal prosecutions (e. All information supplied will be included in the Laboratory Report. In relation to pets. fish and bees. is involved or where public health is likely to be involved or for import and export purposes. Specimens must be accompanied by a Specimen Submission Form signed by the submitter. They may also have analyses performed by the Government Analytical Laboratories. The laboratory fees for most common tests can be downloaded from http://www. and with respect to sealing. aviary etc.asn. If a case is likely to involve police matters or matters controlled by other regulatory authorities. The submitter thereby accepts responsibility for payment of laboratory charges for the tests requested. Because it is often difficult for submitters to specify the precise tests that will be required.nata. SPECIMEN SUBMISSION FORM Field veterinarians should supply all pertinent details of the history. herd. certification and regulatory purposes in accordance with Department policies. the reason for this request and the source of funds (where known) should be clearly and conspicuously stated.au. Laboratory Reports on samples submitted by field veterinarians as routine diagnostic VETERINARY LABORATORY MANUAL LABORATORY CHARGES NSW Department of Primary Industries operates the Veterinary Laboratories on a cost recovery basis. extension. Submitters will normally be invoiced soon after the final report is issued. clinical signs and necropsy findings. native and feral animals and birds.

e.g. RURAL LANDS PROTECTION BOARD and RLPB Property ID The affected property's RLPB (Rural Lands Protection Board) district and its RLPB property ID is required for statistical and reporting purposes. ovine or bovine Johnes disease. Please print clearly. select from the following: male. The space available on the form indicates the level of detail required. as this ensures good photocopies.g. An official Specimen Submission Form should be used with all specimens submitted. The Property ID is the Property Identification Code (PIC). Use the Specimen Submission Key List in addition to the Sample Submission Form if there are insufficient lines on the latter.agric. For the statistics to have meaning. pigs. state clearly. horses. including the property name of the owners.nsw. The number of containers. DISEASE SUSPECTED For Diagnostic cases. select from the following: cattle.gov. Brown. female. Such detailed advice is best emailed to the laboratory for inclusion in the laboratory report. Specimen Submission Forms are supplied in pads of 100 pages. unknown. Long narratives waste your time and ours.au/reader/dasvettesting. Be as concise as possible. give breed or predominant breed in crosses. bees. If you wish the laboratory to hold a specimen for possible testing later. OWNER Provide the normal trading name and full address. These forms provide a check list for information required by the laboratory in these investigations. Alternatively. pens) at the start of the nominated period. should be clearly listed so that the laboratory staff can check to be sure that all the specimens submitted have been received. tail tag number. TESTS REQUESTED Be as specific as possible. NUMBER OF ANIMALS We use the numbers provided to calculate the epidemiological statistics. A. REASON FOR TEST Tick the most appropriate checkbox. breed. deer. give the first two disease possibilities you suspect. This is especially important for regulatory/ movement/ export testing. the Form can be downloaded for printing from the NSW Department of Primary Industries web site at http://www. Provide your email address if you would prefer to receive your results by email. The following provides guidance completing each section of the form: on contact between the laboratory and the veterinarian. swabs and blood samples. For species. 'period prevalence' and 'cumulative mortality'. fish. The same name must be used in all specimen submissions from a property to allow previous submissions from the property to be traced. For sex. SPECIES Include the species. sheep. If others. goats. A.). castrate. Please specify the unit. SPECIMENS SUBMITTED Give a full list of the specimens submitted together with the examinations requested on each. spayed. Your request is equivalent to an order for services and you will be charged for all tests you have requested. a second sheet may be attached. SUBMITTER The full name and address (including postcode) should be printed. e. poultry. For breed. write 'HOLD' beside this specimen. mixed group. It should be clear which test should be applied to each sample. The submitter's telephone and facsimile numbers should also be given to allow VETERINARY LABORATORY MANUAL 2 . Brucella CFT and ELISA. Mia Mia Pastoral Co.NSW Department of Primary Industries' Specimen Submission Form facilitates the orderly collection of this information. or Assessment Number. For more complex investigations. the following conventions should be followed: At risk: The total number of animals or units (eg litters. age and sex of affected animals. Special forms may also be available for special investigations (eg footrot. Specimen Submission forms should be completed using black or blue ballpoint pens. Brown & Sons.

Gross Pathology Representative samples of affected tissue with any adjacent normal tissue. Please specify the period. Further information eg daily mortality or prevalence can be provided in the 'History' section. clinical signs. or date of any previous submissions relating to the condition from this owner should b given. month. and necropsy findings. Parasitology Approximately 30g of faeces for faecal egg count. W04/01762). This allows the duty pathologist to check previous reports. offer more informed comment. we encourage field veterinarians to consult with NSW Department of Primary Industries laboratory veterinarians to decide on the most appropriate sampling and testing strategies. it is better to send too many rather than too few samples. in transport medium. Specimens will be held in the laboratory under appropriate conditions pending the results of initial tests. When investigating difficult or unusual disease problems. Toxicology Approximately 50ml of ingesta. POSTMORTEM FINDINGS An accurate summary of significant postmortem findings will help us provide interpretation of laboratory test results. The pathologist should be given sufficient history and specimens to allow the application of other examinations which may be suggested from the history or lesions. 30ml of fresh chilled tissue eg heart. SPECIMENS (BY DISEASE OR SYNDROME) For details of the most appropriate specimens and tests for a wide variety of animal diseases. Tissues should be in 1cm thick slices in 10x their volume of buffered formalin solution. Virology Full 10ml plain and EDTA blood tubes. NB All specimens should be clearly labelled with the animal identification. Such additional testing will be undertaken only after consultation with the submitter. Biochemistry Full 10ml plain and LiHep blood tube. COLLECTION AND LABELLING OF SPECIMENS SPECIMENS (BY DISCIPLINE) For detailed information on collection and handling of specimens for each laboratory discipline. and maintain continuity between individual submissions from the same owner. faeces or fresh tissue. intestine in a screw-capped container. lifetime.Affected: The total number of animals or units that have been affected (sick or dead) during the nominated period eg year.see Specimens (by discipline) in this Manual. intestinal content. nutrition. with subsections: • Diseases of livestock • Diseases of poultry • Diseases of caged birds • Diseases of bees • Diseases of fish In general. Haematology Full 10ml EDTA blood tube and blood smear. 30 ml of chilled lesion. treatments. HISTORY AND CLINICAL FINDINGS Include details of husbandry. Genetics 50 tail hairs with roots. fluid or tissue eg liver. lung. Dead: The total number of animals or units dead during the same nominated period. The following is a brief guide: Bacteriology Swabs of tissues eg heart blood. NSW Department of Primary Industries does not supply specimen containers or consumables. Histopathology Representative samples of affected tissue with adjacent normal tissue. Serology Full 10ml plain blood tube. see Specimens (by disease or syndrome) in this Manual. VETERINARY LABORATORY MANUAL 3 . PREVIOUS REFERENCES The laboratory reference number (eg. spleen or swabs of lesions or tissues in PBGS.

when exposure occurs. Category A (IATA Packing Instruction 602): An infectious substance which is transported in a form that. Packing Instruction 602 is more stringent than Packing Instruction 650. Examples include Hendravirus. These labels can be obtained from RVLs or from couriers.Care must be taken in the collection and despatch of samples to the laboratory to ensure that you. which are updated annually in the IATA publication ‘Dangerous Goods Regulations’.are available from your Regional Veterinary Laboratory. and exotic pathogens such as FMD virus and Nipah virus. ‘Diagnostic specimens’ is the Proper Shipping Name. personnel handling the specimens during transit and the staff at the Laboratory who have to unpack and handle the specimens are not subjected to any risk of infection. and also the account number of NSW Department of Primary Industries (to charge the cost of freight to the laboratory). NB Virtually all specimens sent by veterinarians to laboratories are transported as ‘Diagnostic specimens’ and must be packed to comply with IATA Packing Instruction 650. *Pre-addressed Consignment Notes carrying the above endorsements. • UN 3373 (Diagnostic specimens) VETERINARY LABORATORY MANUAL . • UN 2900 (Infectious substance. • Within each Category. since carriage by air may be involved. infectious substances are given a UN Number and Proper Shipping Name appropriate for their hazard classification. For postage. Bacillus anthracis (cultures only). are satisfied by packing for carriage by air. NB Virtually all specimens sent by veterinarians to laboratories are transported as ‘Diagnostic specimens’ and must be packed to comply with IATA Packing Instruction 650. IATA PACKING INSTRUCTION 602 (UN2814 INFECTIOUS SUBSTANCE 4 PACKING OF SPECIMENS Submitters are responsible for correctly packing and consigning specimens in compliance with current International Air Transport Association (IATA) regulations. There is a small charge for this service. Incorrect packing may result in refusal by a courier to carry the consignment and/or in legal action. IATA Packing Instruction 650 applies. A UN 3373 DIAGNOSTIC SPECIMENS diamond-shaped label must be applied next to the Consignment Note. IATA divides infectious substances into Category A (IATA Packing Instruction 602) and Category B (IATA Packing Instruction 650). life-threatening or fatal disease to humans or animals. your staff. See IATA indicative list of Category A substances. A number of companies sell suitable packing. but by far the most common outer container is the foam esky enclosed within a rigid cardboard box. in general. affecting animals only). NSW Department of Primary Industries does not supply specimen containers or consumables. A maximum of 1 L within each primary container. Category B (IATA Packing Instruction 650): An infectious substance which does not meet the criteria for Category A. The Consignment Note* must include the following details: • Proper Shipping Name ie ‘Diagnostic specimens’ • Net quantity* and • UN number ie ‘UN 3373’ *Maximum 4 kg of solid or 4 L of liquid material (excluding coolant). See Guidelines for Packing Diagnostic Specimens below for practical advice on packing to comply with IATA Packing Instruction 650. other regulations apply but. recyclable specimen containers or eskies will be returned to submitters. On request. For carriage by road. *Proper Shipping Name. is capable of causing permanent disability. affecting humans*). IATA PACKING INSTRUCTION 650 (UN 3373 DIAGNOSTIC SPECIMENS) This instruction covers the labelling and packing of Category B infectious substance as defined by IATA. • UN 2814 (Infectious substance.

Outer container (usually an esky) i.au/agriculture/vetm anual/contact . iii. The following notes are provided to assist submitters. the samples may be sent by that laboratory to another appropriate Departmental testing centre. IATA Packing Instruction 602 is more stringent than 650.it must be sealed watertight. should be consulted for advice on specimen packing requirements. Tighten the lids of all primary specimen containers. GUIDELINES FOR PACKING DIAGNOSTIC SPECIMENS (IATA PACKING INSTRUCTION 650) ‘Dangerous Goods Regulations’. Put Specimen Submission Form(s) into a separate plastic bag. If the esky is foam. should be submitted to the Regional Veterinary Laboratory which services your region. iv. Primary containers i. available from your Regional Veterinary Laboratory. and requires: • Person packing to have been trained by an IATA-approved trainer within the past two years. Seal all primary containers from each property/problem into a VETERINARY LABORATORY MANUAL watertight secondary container with sufficient absorbent material to absorb all fluid from the primary container(s). TRANSPORT OF SPECIMENS Specimens (other than those for export testing). Check that the specimens tally with those identified on the Specimen Submission Form. For histopathology containers. Estimate the total net weight of specimens and record this on the Consignment Note. Complete the Consignment Note and stick it onto the outer container. Call Customer Service (1800 675 623 if you need the Account Number of NSW Department of Primary Industries for a particular courier (to charge the cost of freight to the laboratory). Bacillus anthracis (cultures only). wrap together between stiff cardboard. A sturdy plastic bag will suffice as a secondary container . and exotic pathogens such as FMD virus and Nipah virus. iv. tape the lid to prevent loosening.nsw. See IATA indicative list of Category A substances. For glass slides. cooler bricks and tight packing into the outer container (usually an esky).AFFECTING HUMANS or UN2900 INFECTIOUS SUBSTANCE AFFECTING ANIMALS ONLY) This instruction covers the labelling and packing of Category A Infectious Substance as defined by IATA (see above). Pre-addressed Consignment Notes. NB Virtually all specimens sent by veterinarians to laboratories are transported as ‘Diagnostic specimens’ and must be packed to comply with IATA Packing Instruction 650.dpi. seal it into a cardboard box. Put specimen secondary container(s). ii. Crumpled newspaper will suffice for tight packing. published annually by the International Air Transport Association (IATA). Stick a UN 3373 DIAGNOSTIC SPECIMENS diamond-shaped label adjacent to the consignment note. Generally specimens should not be sent by overnight courier on Friday or at the 5 . iii. This may be another RVL or one of the Central Laboratories at EMAI. ii. Clearly label each primary specimen container or slide with the animal identification. ii. include this Account Number NB Suspect ANTHRAX submissions must have a clear warning under the lid of the outer packing. It includes those agents that cause serious communicable disease. A map showing the location of laboratories is available at http://www. These labels are available from RVLs or from some couriers. Secondary container(s) i. Depending on the tests specified. Indelibly label the secondary container with the owner's name. Menangle. Examples include Hendravirus. iii. • Completed Shipper’s Declaration for Dangerous Goods form with the package.gov. Specimen Submission Form(s). This warning must be on top of samples and not packed in with the samples.

contact the Animal Disease Watch Hotline 1800 675 888 or the local Senior Field Veterinary Officer. NSW Department of Primary Industries encourages veterinarians to submit samples to their laboratories by offering some 'free' testing. ii. All costs associated with freight and carcase disposal will be borne by the submitter unless prior arrangements have been made. Submitters can therefore charge the cost of freight to the NSW Department of Primary Industries’ Account Number on the Consignment Note. This passively-collected data is supplemented by data collected while testing healthy animals for export or interstate movement and by active surveillance programs funded by industry or by the Department.gov. Animals should be placed in the container and delivered to the terminal or railway station shortly before the estimated time of departure. no fee will be charged for tests conducted at a NSW Department of Primary Industries laboratory to exclude an endemic notifiable disease.au/agriculture/vetm anual/submission/disease-surveillance. It is the responsibility of the sender to meet with the requirements not only of the transport authority. all testing to exclude that disease and to establish an alternative diagnosis will be free of charge to the submitter. DISEASE SURVEILLANCE NSW Department of Primary Industries collects epidemiological data on the prevalence of animal disease by monitoring the presenting problem and result of testing of material submitted to its laboratories. NOTIFIABLE DISEASES A full list of the currently notifiable diseases is available at http://www. Weekend delivery attracts a heavy surcharge from all courier companies. For emergency (exotic) disease. We will determine the feasibility of the operation and make arrangements for receiving the animals. COURIERS Most submissions are sent to our laboratories by overnight courier.nsw. For notifiable diseases (including emergency diseases) and for mortality investigations. The cost of transport of submissions to the laboratory is included in the test charges. This data is used to satisfy the requirements of our trading partners and to direct research and extension efforts of our staff. to lie down fully extended and to turn round. but also with the requirements of welfare of the animal during transport.weekend. FORWARDING LIVE OR DEAD ANIMALS The laboratory should be consulted before live or whole dead animals are forwarded. For endemic diseases. Veterinarians and livestock owners have an obligation under various Acts of Parliament. submission of specimens to a NSW Department of Primary Industries' laboratory accompanied by a fully completed Specimen Submission Form nominating the suspected disease is a suitable form of notification. call the laboratory to discuss options. For emergency (exotic) disease. Full details of the eligibility criteria have been circulated to veterinarians in NSW. Suitable bedding should be provided. NSW Department of Primary Industries has contracts or agreements with several large courier companies as well as with local companies. Some companies offer same day delivery.dpi. Generally. left confined for long periods awaiting or during transport. The containers should be large enough for the animal to stand. iv. Commercial couriers may use either road or air transport and specimens should therefore be packed in accordance with the more stringent IATA requirements for air transport. iii. The following should be used as guidelines: i. to report any suspected occurrence of these diseases to NSW Department of Primary Industries staff. Before calling your courier to pick up specimens on a Friday. Call the laboratory to discuss transport options before contacting a courier company. Animals should not be VETERINARY LABORATORY MANUAL 6 . Containers must meet the requirements of the transport authority.

iii.cfm. See also Transmissible spongiform encephalopathy in this Manual. fax (02) 8334 7430] of their intent to export. ix. where necessary. It is particularly important for export testing that samples of good quality and adequate quantity be submitted and that they be correctly labelled.animalhealthaustralia. (These values are GST inclusive) Submission Form.au/pr ograms/adsp/adsp_home.. Veterinarians and owners receive incentive payments. When using this subsidy. at least four (4) weeks prior to the commencement of testing. Menangle or his/her appointed nominee should be advised of the expected dates of sampling and shipment of the animals prior to the submission of the specimens. • All owner and property details must be provided on a fully completed Specimen Submission Form. viii. This information must be included on the Specimen vi. For all overseas export testing the samples should be forwarded directly to: Officer in Charge Regional Veterinary Laboratory Elizabeth Macarthur Agricultural Institute Woodbridge Road MENANGLE NSW 2568 The Regional Veterinary Laboratory and the Central Laboratories at EMAI. Intending exporters (owners and/or agents) must advise the Animal Quarantine Inspection Service (AQIS) [phone (02) 8334 7434.com. The Officer in Charge of the Regional Veterinary Laboratory. Full legal animal identification details should be cross-matched to sample identification and documented. vii. ii. and to prepare quotations for the cost of laboratory services. v. Field veterinarians should check to ensure this requirement has been met. This will allow the laboratory to check on any changes in requirements. NB Foetuses and neonates up to one day of age are not eligible for this subsidy. The vendor of the stock should be listed as the owner of the stock rather than the agent or shipping company. To qualify for this subsidy: • The animals involved must be from a commercial livestock herd or flock. This allows different lots to be traced. Horse mortalities are eligible. The veterinarian submitting material is totally responsible for determining the tests required. MORTALITY INVESTIGATIONS NSW Department of Primary Industries subsidizes laboratory fees for the investigation of mortalities in livestock. fax (02) 8334 7430). Specific requirements for testing of stock for each overseas export destination can be obtained from the Animal Quarantine Inspection Service (AQIS) [phone (02) 8334 7434. The RVL Menangle co-ordinates 7 VETERINARY LABORATORY MANUAL . submitters pay the first $80 of the laboratory fee and NSW Department of Primary Industries up to $140 of the residual. otherwise there can be no guarantee that testing will be completed and certification provided to allow export to proceed on schedule. The name of the agent or shipping company involved should be supplied on the Specimen Submission Form with telephone/facsimile numbers.NATIONAL TSE SURVEILLANCE PROGRAM (NTSESP) Full details of this program are available at http://www. SPECIAL REQUIREMENTS FOR TESTING OF STOCK FOR OVERSEAS EXPORT The following requirements should be fulfilled: i. Menangle offer almost all required export tests for livestock. The laboratory charges for this testing are charged to the NTSESP. For poultry more than 100 birds must have died. when animals are being purchased from several properties. iv. The name of the official veterinarian supplying the information on export testing requirements should be advised and stated on the Specimen Submission Form. Veterinarians are encouraged to submit specimens from adult sheep and cattle with progressive neurological disease.

This will expedite testing. Nomination of tests to be performed is the responsibility of the authorised official submitting veterinarian. v. Full legal animal identification must be documented. and that they be correctly labelled. VETERINARY LABORATORY MANUAL 8 . SPECIAL REQUIREMENTS FOR TESTING OF STOCK FOR INTERSTATE MOVEMENT i. vi. send specimens to the relevant Regional Veterinary Laboratory for the district of origin of the stock. Those tests unavailable at Regional Veterinary Laboratories will be undertaken at EMAI following referral of specimens from the Regional Veterinary Laboratories. and Wollongbar for export certification. the Officer-in-Charge of the local Regional Veterinary Laboratory or his/her appointed nominee should be advised of the expected dates of sampling and ii. The vendor of the stock should be listed as the owner of the stock. transport of the animals. In cases where large numbers of animals are to be tested for interstate movement (i. prior to the submission of the specimens. This allows different lots to be traced. when animals are being purchased from several properties. iv. certain tests can be performed within Regional Veterinary Laboratories at Orange. Alternatively.e. For interstate movement. Advice on requirements for interstate movement should be obtained from the District Veterinarian of the local Rural Lands Protection Board. rather than the agent or transport company. iii. It is particularly important that samples of good quality and adequate quantity be submitted.most export testing in NSW and can be contacted by all field veterinarians directly for enquiries and assistance. greater than 200).

25 ml • Bijou Macartney . heparin and EDTA Vacutainer needle holders and needles (18G) Sterile containers (100 ml) Sterile bottles • 1 oz Macartney . Xylocaine 2%. Rompun 2% ii.5 cm blades Handsaw 30 cm blade and replacement blade Scissors • Mayo straight 16 cm • Mayo straight 14 cm • Double sharp 11 cm fine • Pointed/1 rounded end and knob (for gut running) 6 Euthanasia 7 Necropsy • • • • • • • • • • • • • • • • VETERINARY LABORATORY MANUAL 9 .5 cm wide straight blade • 20 cm skinning 2. disposable ii.5 ml Swabs Transport media • Amies charcoal transport medium in bottles of commercial swab packs • Phosphate buffered glycerol saline (PBGS) in bottles Microscope slides (and spreaders for blood films) Syringes (1-20 ml) and needles (14-26 G) Scalpel handle and disposable blades Scissors Non-sterile faecal containers (50-100 ml) Vaginal mucus pipettes Ethanol/iodine for skin asepsis Biopsy instruments and small bottles of fixative Rifle and ammunition Euthanasia solution and syringe/needle Killing knife Knives • 20 cm skinning 2.5 cm wide curved blade • 15 cm boning knife pointed straight Steel . post mortem gloves or gauntlets • Post mortem apron (optional) • Towel and soap • Wet weather gear 2 Animal Husbandry • Halter • Nose grips • Bleeding choke rope • Twitch Pig handler • Drugs i. etc 3 Clinical • Stethoscope • Thermometers • Obstetrical gloves 4 Clinico-pathological • Nitrate and cyanide test 5 Clinical Specimens • • • • • • Vacutainers .30 cm Butcher's Footrot secateurs • 7.plain.CHECKLIST OF EQUIPMENT FOR CLINICAL AND NECROPSY EXAMINATIONS Suggest Checklist for Clinical and Necropsy Examinations 1 Personal • Rubber boots • Overalls • Gloves i.

5 ml VETERINARY LABORATORY MANUAL 10 . 4 • Scalpel blades • No. dry ice or portable car fridge • Serum storage plastic disposable tubes .g. 20 • Rib cutters • Meatsaw/hacksaw • Cleaner/hatchet • Buckets . or markable slides and pencil • Histopath jars with 10% neutral buffered formalin • Large 500 ml • Medium 250 ml • Small 125 ml • Special fixatives (e.Suggest Checklist for Clinical and Necropsy Examinations • Bone forceps • Toothed forceps • 20 cm • 18 cm • 13 cm • Scalpel handles • No.1 mm thick • large • medium • small • Rubber bands • Microscope slides and diamond pencils.suitable container for contaminated 'sharps' 10 Clerical • Clip board • Postmortem and Specimen Submission Forms • Specimen collection handbook • Pencil • Marking pens • Black biro • Adhesive labels • String-tie labels 11 Storage • Large metal esky with a blood vacutainer box and small foam esky inside • Frozen cold bricks.plastic (2) • Trays • plastic (large) • plastic (small) • Sterile scissors and forceps for virological tissue samples 8 Pathological • Sterile containers Specimens • 100 ml • 25 ml • 5 ml • Plastic bags 0. Bouins) if required • Ball of string 9 Decontamination • Container and 20 litres water /Disinfection • Disinfectant – one litre Lysol or similar • Nail scrubbing brush (for cleaning instruments) • Long handled scrubbing brush (for cleaning boots) • Roll paper towel • Plastic garbage bags . 24 and/or No.

swab/charcoal transport medium for more sensitive organisms. so that recovery of the organism alone may not necessarily be significant. wide-mouthed. A minimum bag thickness of 0. in association with pathological changes.au/agriculture/vetm anual/specimens-bydiscipline/bacteriology/02-bull-vd-coll.0 cm in length. such as swab/transport medium packs for general purposes. Some testing involving special stains or antigen testing may require dry swabs or specific transport conditions COLLECTION OF SPECIMENS Specimens should be collected aseptically and submitted promptly in individual sterile non-leaking.pdf Plastic pipettes made from polystyrene or polypropylene with suggested dimensions of external diameter 9. There are different transport mediums available commercially. which are inserted into a semisolid transport medium (such as Stuart Transport Medium or Amies Charcoal Transport Medium) particularly for fastidious bacteria. They should be fitted to 90 ml firm rubber bulbs for suction while scraping.4 mm. Swabs Swab transport systems for the recovery of organisms are available commercially.g. e. before submission to the laboratory. the entire lesion or organ should be submitted. If small. The primary receptacle must be leakproof and not contain more than 500g. the bag should be packed inside a second bag which also contains sufficient absorbent material to absorb fluid should the inner bag leak and securely sealed as above. Many pathogenic organisms are present in normal animals. These are available commercially. gloves or fragile containers when submitting material for bacteriological examination. screwtopped. is bevelled. Slides with frosted ends are preferred for labelling purposes. but only a few are commonly pathogenic. etc. wrapped in tissue/paper with sufficient padding. STANDARD CONTAINERS AND EQUIPMENT Sterile plastic bottles and jars These are available commercially in a range of sizes from about 10 to 100 ml. discharges. Plastic bags may be suitable.nsw. the other end. swab/transport systems modified for anaerobe recovery. If the lesions are large or widespread. Pipettes for collecting preputial scrapings from bulls See http://www.1 mm should be used. straight except for a bend at the end that is held by the operator during collection. Pipettes for collecting vaginal mucus Polystyrene artificial insemination pipettes. E coli. NB Do not use non sterile containers. Ensure slides are packed such that they don’t break in transit eg slide holder. small lesions. cm long x 2mm internal diameter and 2 ml capacity. 61.g. approximately 50. submit a portion of the affected tissue containing the lesion and surrounding area. horns or other sharp objects may puncture the bag. They are used for fluids. plastic container. there are many serotypes.5 mm. Plastic bags For larger specimens of organs or foetuses. or plastic jars.g. e. In other cases e. Plastic bags must not be used if bone.gov. Clostridium perfringens in intestinal contents.dpi. Some commercially available slides are not washed and are not suitable for making satisfactory smears. screw-topped. Microscope slides Use slides of standard size and ensure they are clean.SPECIMENS (BY DISCIPLINE) BACTERIOLOGY A bacterial disease can only be diagnosed when a pathogenic organism can be demonstrated. using molecular techniques or in tissue sections. Alternatively an aspirate of the lesion can be taken and transferred to a small sterile container and submitted 11 VETERINARY LABORATORY MANUAL . culture. which contacts the preputial surfaces. Aspirates submitted in syringes with needles will not be examined. They consist of plain cotton wool sterile swabs. either on smear. internal diameter 6. sterile 30 ml universal containers. All bags should be securely sealed by ziplocking or double folding and application of stout rubber bands or equivalent. Milk sample bottles Sterile 30 ml Macartney bottles or any equivalent sterile.

spleen. If a parasitological examination is also required. NB.nsw. non-leaking containers. Intestinal contents Should be expressed into sterile. (preferred collection method) Rectal swabs. heavily impregnated with faeces. should be submitted in transport medium such as Stuart Transport Medium or Amies Charcoal Transport Medium.gov.. Aborted foetuses and foetal membranes For foetuses from smaller animals (e.pdf Selective transport medium for T foetus (InPouch TF). a smear(s) should be made and submitted with the swab. Stuart Transport Medium is unsuitable for contagious equine metritis (Taylorella equigenitalis) transport if the swabs are to be processed more than 24 hours after collection. a postmortem examination should be conducted and the appropriate specimens should be collected aseptically into sterile jars (see ‘Abortion’). Milk samples Wash the udder and the teats thoroughly and dry with a paper towel. about 10 ml milk squirted into a sterile 30 ml Macartney bottle or sterile 30 ml plastic universal container held nearly horizontally. Faecal samples and rectal swabs Collect approximately 30 g faeces direct from the rectum into a sterile. Preputial samples should not be frozen or refrigerated. Do not overfill the container. a duplicate sample should be collected. pigs) and smaller foetuses from the larger animals. A number of smears can often be made on each slide. Portions of the placenta or cotyledons should also be taken and fixed in buffered formalin prior to despatch. In the case of exudates. are generally inferior to the above. together with instructions for the collections and despatch of specimens will be forwarded from the laboratory on request. Swabs of intestinal contents should be placed in a transport medium (such as Stuart Transport Medium or Amies Charcoal Transport Medium).dpi. sheep. Impression smears of foetal membranes should be taken prior to despatch of specimens. avoiding gross contamination of the sample or the container (preferred collection method).g.If a septicaemia/bacteraemia is suspected. submit portions of liver. In the case of the large foetuses which cannot be delivered to a laboratory. widemouthed. Swab the teats with 70 per cent alcohol. Smears of intestinal mucosa and pathological lesions Firmly smear suspect area with slide. Milks must be refrigerated if transport is slightly delayed.au/agriculture/vetm anual/specimens-bydiscipline/bacteriology/02-in-pouchguide. heart blood and lung. Always leave one (frosted) end of the slide clean for handling and labelling. see the section ‘Johne's disease’. Code the samples numerically. AIR DRY and wrap separately or place in slide protective holders for transport (commercially available). The samples must be submitted chilled using either crushed ice or cooling bricks in an insulated container. Keep at temperatures between 18oC and 30oC. Swabs Are generally inferior to the above. Avoid touching the mouth of the bottle with the teat. For Johne's faecal culture. for culture should be submitted as soon as possible in transport medium (such as Stuart Transport Medium or Amies Charcoal Transport Medium) is preferred. The first couple of squirts should be discarded unless used for a field test and. In the case of exudates. goats. a smear(s) should be made and submitted with the swab. widemouthed non-leaking container. Smears should be clearly labelled. Preputial material from bulls Preputial scrapings submitted unchilled in selective transport medium for T foetus (InPouch TF) See http://www. . or frozen if transport is delayed more than 2-3 days. Semen samples VETERINARY LABORATORY MANUAL 12 . the entire foetus together with the foetal membranes should be enclosed in separate plastic bags and submitted chilled. in separate containers.

This will ensure the minimum growth of contaminants.25 ml of undiluted formalin or 1.dpi. Otherwise.5 ml of 10% formalin. Vaginal mucus samples Mucus can be collected from the ventral fornix of the vagina and the external os of the cervix by guiding a plastic artificial insemination pipette by hand (per rectum) and applying GENTLE suction to the external end. on dry ice is recommended. (Preferred method): Vaginal mucus samples are collected for demonstration of C fetus subsp venerealis antibodies by BVC ELISA test. should be kept chilled (2-8oC) but not frozen. For isolation of Campylobacter.au/agricultur e/vetmanual/specimens-bydiscipline/bacteriology/01-bvc-elisaguide. Consult with your courier if you intend to use dry ice for submission of samples. all samples for bacteriological examination. PBST diluent and instructions for collection of mucus samples are available from the laboratory. consult with your Regional Veterinary Laboratory. direct inoculation to Campylobacter transport media (CTM) is ideal. then sealed pipettes from which air bubbles are excluded should be used (Atmospheric oxygen is lethal to Campylobacter fetus). but should be processed by the laboratory within 72 hours of collection. preserved with 0. Urine samples For routine cultural examinations to be meaningful. From 0. VETERINARY LABORATORY MANUAL 13 . Specimens are better held refrigerated until the sender is sure the transport to the laboratory will not be delayed. Specimens best stored at room temperature (not refrigerated) The following samples should be stored o o and transported between 18 C and 37 C (ie not refrigerated or frozen): • Samples submitted in Campylobacter Transport Medium (CETM) • Samples submitted in Trichomonas Transport Medium (In Pouch) • Swabs submitted in Amies Charcoal medium for Contagious Equine Metritis (CEM) • Swabs submitted in Amies Charcoal medium for or Atrophic rhinitis. for leptospire morphology submit a separate urine sample of at least 20 ml. See http://www.5 to 1. Samples collected using an artificial vagina are often grossly contaminated unless they are cultured shortly after collection. Vaginal mucus is collected using a plain. When direct examination for leptospires is required. submit a fresh urine specimen specifically for leptospiras motility examination at the laboratory within 20 minutes of collection. urine samples should reach the laboratory within a few hours of collection. Plastic pipettes can be sealed adjacent to the mucus using a pair of pliers previously heated in a flame. Brachyspira). Insulated containers with frozen icebricks or plastic bags of crushed ice should be taken in the car to all investigations and specimens placed therein as soon as possible. The swabs. Consult with your courier. To diagnose bovine venereal campylobacteriosis (BVC) in abortion or infertility cases: i. except smears. STORAGE OF BACTERIOLOGICAL SPECIMENS PRIOR TO DESPATCH In general. If culture is required. obligate anaerobic bacteria (including Dichelobacter nodosus) have special requirements. If there are likely delays before or during transport.gov. Chlamydia. Samples of vaginal mucus for bacteriological examination for Campylobacter fetus should be submitted on dry ice in an insulated container. ii. Attempts to collect large volumes by repeated ejaculation increase the risk of contamination. Bacteria with particular requirements Mycoplasma.0 ml is sufficient for bacteriological examination. Spirochaetes (including Leptospira. If specimens for Campylobacter culture will not reach the laboratory within 6-8 hours. Delays in transport over weekends and public holidays must be considered. sterile cotton swab and placing the cotton swab end into phosphate buffered saline plus Tween (PBST) for transport. PBST and instructions are provided by the laboratory. specimens must be chilled.pdf.Must be collected aseptically avoiding contamination by preputial material. from the time of collection until they are received in the laboratory.nsw.

chilled. COLLECTION OF SPECIMENS Blood and serum samples Blood samples should be collected aseptically into the appropriate 10 ml vacuum tube.. Separated sera or plasma must be placed into 5 ml BIOCHEMISTRY NSW Department of Primary Industries’ laboratories do not carry out any biochemical analysis. In some cases it is more appropriate to apply a test on fresh samples in the field rather than submit samples to a laboratory. and disposable plastic containers have eliminated this problem. total protein. • Serum vials: 5 ml vials or bottles with leak-proof closures. hair. • The specimen advice form is best sealed in a plastic bag to avoid contamination or damage from water. without anticoagulant: For blood and serum samples.• Tissues for fungal culture (temperatures of less than 15oC can be detrimental to fungal survival). albumin and enzyme levels. • Specimens for transport from cases of suspected tuberculosis or anthrax should always be submitted individually in containers separate from all other specimens and in accordance with IATA requirements for infectious substances (Packing Instructions 602).e. must be avoided. or at room temperature. Haemolysis will seriously interfere with the level of serum magnesium. phosphate. etc. Chilling should also continue in transit.for tissues. sera should be separated from the clot as soon as possible and frozen or chilled in 5 ml serum vials. jars. e. • Bottles. rather than the same test applied in the laboratory 24 to 48 hours later. Chilling should also continue in transit. then forwarded chilled to the laboratory. free of all cells and then kept chilled or frozen. Care should be taken to avoid haemolysis (see "Serology . • Check to ensure that specimens are submitted at the temperature appropriate for the disease condition.Avoiding Haemolysis of Samples").. Check with your courier. Pack such that on opening the outer packaging. Where whole blood will not arrive at the laboratory within 24 hours of collection. The plasma should be removed. plasma should be separated by centrifugation as soon as possible and preferably within 4 hours of collection. Sera for Vitamin A or E analysis must be protected from heat and light. Vacuum blood tubes and needles. The blood sample should be collected in the appropriate blood vacuum tube. DESPATCH OF SPECIMENS See section ‘Packaging and sending specimens to the laboratory’. Plasma for Vitamin A or E analysis must be protected from heat and light. faeces. Separated sera or plasma must be placed into 5 ml containers wrapped in foil. • Blood vacuum tubes (10 ml) with EDTA or heparin: For plasma samples.g. etc. siliconized. i. Contamination of the sample by soil. VETERINARY LABORATORY MANUAL 14 . where commercially available kits will provide a more reliable answer on the spot. Where whole blood will not arrive at the laboratory within 24 hours of collection. all such testing is outsourced to laboratories that are accredited by NATA to conduct biochemical tests. urine analysis. when forwarding specimens for bacteriological examination the following procedures should be adopted: • All specimens for transport should be packed in accordance with IATA requirements for diagnostic specimens (Packing Instructions 650) so as to prevent leakage and risk of accidental exposure of personnel handling the container. a note advising ‘Suspect TB’ or ‘Suspect Anthrax’. STANDARD CONTAINERS AND EQUIPMENT Contamination of glassware and other collection equipment by traces of the element of interest has been an issue in the past. as listed for Bacteriology. organs. Plasma samples At least 2 ml of plasma should be submitted frozen in 5 ml screw capped containers. plastic bags . Standard containers for Biochemistry are: • Blood vacuum tubes (10 ml). In addition.

then duplicate samples as required for the other examinations should be submitted in separate containers.pdf 15 .g. Paper envelope or pre-labelled ‘ziplock’ plastic bags For tail hairs. GUIDE TO INTERPRETATION OF BIOCHEMICAL PARAMETERS IN SHEEP AND CATTLE Normal values are supplied by the laboratories to which we outsource and are included in our reports to submitters. bacteriology. and post to EMAI.dpi.gov. Firmly grasp the knotted hairs and pull with a quick action. pathology. Muscular degeneration. always ensure there are several ice bricks to ensure samples arrive at least chilled: • Always seal the specimen advice form in a separate plastic bag. Care must be taken to avoid contamination by soil. Refer to relevant section of the manual.nsw. If other examinations are required. Tie a knot in the hair shafts approximately one quarter of the distance from their proximal end. e. cut off and discard the distal half of the shafts. breeding data and clinical findings. nutritional Selenium deficiency Serum enzymology Vitamin A and E Vitamin B12 GENETICS The field veterinarian should check the Specimens (By Disease or Syndrome) section of this manual for details on the specimens required for diagnosis of the specific disease suspected. Where DNA based tests have been developed for genotyping at a defined locus the preferred sample is hair roots. Suggested normal values for some analytes are included under specific diseases in the section ‘Specimens (by disease or syndrome)’ of this manual.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot. Ensure roots are present at the proximal end of the hair shafts. diagnosis may be possible by histological examination of suitable tissues and findings used to complement historical observations. ruminal or intestinal contents when collecting tissue for biochemical analysis. Where DNA tests are not available. STORAGE OF SPECIMENS PRIOR TO DESPATCH Samples of tissues must be chilled or frozen as appropriate until despatch. however the tests will be significantly more expensive than with those that exploit hair roots as a source of DNA. Tissue samples At least 50 g of tissue should be submitted frozen in a screw capped container. See http://www.containers wrapped in foil. To prevent frozen samples from becoming heated during transit. wrap in foil or brown paper and submit chilled to the laboratory. Select hairs that are free of visible faecal contamination. DESPATCH OF SPECIMENS Specimens should be forwarded in an insulated container with an icebrick. If desired. See: Acetonaemia Copper deficiency Hypomagnesaemia Hypocalcaemia Hypophosphataemia VETERINARY LABORATORY MANUAL STANDARD CONTAINERS AND EQUIPMENT Vacuum blood tubes (5mL or 10mL. COLLECTION OF SPECIMENS Hair Tease out 20 to 25 hairs growing at the distal end of the tail. Where values supplied by laboratories differ from these suggested values. with anticoagulant EDTA or heparin) For blood samples. Tissues for vitamin A or E analysis These should be protected from heat and light. Place the shafts (with roots) in an envelope labelled with the animals identity. DNA can also be isolated from anticoagulant-treated blood and from semen. then forwarded chilled to the laboratory. the former should be used. Different organs must be submitted in separate containers. faeces. Only with prior arrangement with the laboratory can other tissue samples be the utilised as the source of DNA for routine tests. The ‘Serum enzymology’ section includes information on enzyme changes see in various conditions.

NB Do not freeze samples being submitted for either gross or histopathological examination GROSS PATHOLOGY Provided that the field veterinarian can be sure that the specimens will be delivered to the Regional Veterinary Laboratory promptly. When the necropsy is done in the field and a laboratory opinion is sought on gross changes detected. Refer to Guidelines for Packaging Specimens for more detail. Submission of the animal. dry hairs are stable at ambient temperature for an extended period of time. All animals either live or dead. When live or dead animals are submitted. STANDARD CONTAINERS AND EQUIPMENT Specimens for gross pathology should be submitted in glass or plastic screw-top containers or strong plastic bags. should be considered when there is a: • High mortality or morbidity from an unknown cause. The laboratory should be advised by phone that the animal is being VETERINARY LABORATORY MANUAL HISTOPATHOLOGY In general. Use sufficient icebricks to ensure samples arrive at least chilled. submitted to the laboratory must be accompanied by the appropriate specimen advice form with all relevant details. Always seal the specimen advice form in a separate plastic bag. live or dead. then a large portion of the tissue or organ should be submitted chilled but not frozen. containing the lesion and adjoining normal tissue for comparison.5mL of raw semen in a serum vial. • Continuing problem and previous examinations have not established a diagnosis. Do not freeze blood samples. from the coccygeal vessels or the jugular vein. together with a good history and a description of the clinical and necropsy findings. STANDARD CONTAINERS AND EQUIPMENT (Neutral) Buffered Formalin Buffered formalin solution is the recommended fixative because of its stability. Specimens other than hairs should be forwarded in an insulated container with an icebrick. Label each tube with the identity of the subject. Clean. COLLECTION OF SPECIMENS FOR GROSS PATHOLOGY When submitting whole animals. delivered. longer life of preserved tissues and 16 . If material is submitted for macroscopic examination. the containers used should prevent contamination of the environment with possible pathogens. Blood Fill evacuated blood tubes. there are advantages in submitting the whole animal or whole affected organs.Use of forceps or artery clamps may facilitate collection of suitable samples from newly-born calves. Semen A semen straw or 0. STORAGE OF SPECIMENS PRIOR TO DESPATCH Tissue samples should be kept chilled from the time of collecting until they are received at the laboratory. Clearly state the breed of the subject and DNA test required. STORAGE OF SPECIMENS AND DESPATCH OF SPECIMENS Samples of tissues other than hairs must be chilled until despatch. ensure that they are typical of the syndrome being investigated. the histopathologist will only be able to offer comment on the significance of lesions when an appropriate range of specimens has been submitted. to end of draw. small portions in buffered formalin should also be taken and submitted for histopathological examination. Immediately after drawing the sample ensure mixing of anticoagulant and blood by repeated but gentle inversion of the tubes. Any dead animal should arrive in time to allow a post mortem examination on the same day.

g. in plastic buckets) for at least 48 hours. If the brain cannot be submitted whole.P.com. They should include part of the lesion and adjacent apparently healthy tissue. Each spinal nerve root is then severed as it is exposed.animalhealthaustralia. the spinal cord should be lifted by grasping the dura with tissue forceps. It should be used neat in this formulation. Organs greater than 1 cm thick must be sectioned.0 g 6. it may be cut transversely into two or three pieces. Brain There are several methods of exposing the brain for removal.2H2O Na2HPO4 * 100 ml 4. It is prepared as follows: Commercial Formalin NaH2PO4. to keep the cord segments connected. Gastrointestinal tract and other tubular organs VETERINARY LABORATORY MANUAL 17 . straight segments.a negligible fixation deposit.5 cm (and no more than 1 cm) thick. then a small block of the organ (sliced 0. To prevent fixation of the cord in a curled position. Tissue for fixation should be sliced 0. Compression of brain surface resting on the base of the container can be avoided by adding neat formalin until the brain floats. The dura is then opened longitudinally in the dorsal midline and the cord and dura submitted whole in a large container of buffered formalin. then submitted in a large container of fresh buffered formalin solution. Do not cut the brain longitudinally. Always submit an adequate range of tissue specimens. It is essential to avoid distortion of the brain during fixation. The cord will then be fixed in short.5 g Ensure that the mucosal surface of tubular organs (eg gut. Water to 1 litre * Commercial formalin is Formalin B. leaving a part of the dura intact at each transection site. Don’t squeeze large tissue samples into containers. Open the dural to allow better penetration of the fixative. Preserved tissue should be submitted in at least 10 times its volume of fixative. Containers These must have a tightly fitting lid to prevent leakage. After 24 hours fixation. uterus.cfm) It is preferable to submit the brain whole in a wide mouth container with 10 times its volume of buffered formalin solution. Other fixatives Other fixatives for special purposes are available from the laboratory. Decomposing or frozen tissues are not suitable for histopathological examination Organs Organs smaller than 1 cm in thickness can be fixed whole. Transect the cord at 10 to 20 cm intervals. Spinal cords After exposure. bovine and equine brains should be immersed in at least 6 litres of buffered formalin solution (e. Handle only the edges of delicate tissues so that histological detail is not lost. 3438% formaldehyde. COLLECTION OF SPECIMENS Tissues collected at necropsy should be preserved immediately.au/pr ograms/adsp/adsp_home. Containers must have sufficiently wide mouths to allow tissue to be easily withdrawn after fixation. bladder) is exposed by excision of the wall before immersion in fixative. Ideally. for histopathology. Always cut transversely so that all brain sections can be examined (grossly and histologically) for bilateral lesions.5 cm and no more than 1 cm thick) should be fixed in buffered formalin at the time of collection. Convenient methods for large animals (eg longitudinal and transverse craniotomy) are used in the National TSE Surveillance Program (http://www. If fresh organs are being submitted for bacteriological or gross pathological examination. Buffer salts are available commercially in prepared packs. the formalin solution can be changed to hasten fixation.

Spreaders can be re-used if cleaned and dried after each use. Blood films can be submitted in the same container as chilled specimens. pasture availability. • DrenchTest kits include 50 containers for one control and four test drench groups. Vacuum blood tubes (with anticoagulant) For heartworm antigen and microfilaria tests in dogs (generally for export purposes). not frozen. collect blood in EDTA (purple top) tubes. PREPARATION OF BLOOD FILMS i. whilst being held prior to despatch and during transport to the laboratory. 70 ml) screw top containers for insects. Use a glass slide as the spreader The spreading edge must be smooth (i. aliquots of gut washings. NB Do not chill blood films or expose them to formalin vapour. VETERINARY LABORATORY MANUAL 18 . • Horse WormTest kits are suitable for 5 horses. They should then be air dried. parasites. stage of pregnancy and effects of lactation. Starvation will increase egg counts and inappetence may cause the count to increase by up to 30 to 40 times. Label at the thick end with the owner's name and animal ID (if necessary) using a lead pencil. Wave the slide quickly in the air to dry it. Grasp the spreader with thumb and middle finger using the index finger to put light pressure on the spreader. Avoid contact between the blood sample and icebricks. no chips out of it) and clean. • DrenchRite® kits for bulk faecal collection allow farmers to test for resistance with a single muster. provided the blood films are securely packed and well wrapped for insulation against chilling. skin scrapings. Place the spreader beyond the drop and at 45o to the slide. Quickly and smoothly push the spreader right to the end of the slide at the 45o angle. host resistance. iv. Place a small drop of blood (thoroughly but gently mixed) in the centre of the slide about 1 cm from the end.e. haemoglobin estimation and derived parameters. STANDARD CONTAINERS AND EQUIPMENT Jars Plastic (5 ml. Faecal egg counts are influenced by faecal consistency and bulk. free of dirt and grease. grazing rotation. The history should provide details of recent anthelmintic treatments including drench resistance status. Parasite Faecal Collection Kits Collection kits are available from your Regional Veterinary Laboratory: • WormTest kits contain 10 containers for monitoring a single mob or grazing group in a herd. Diarrhoea will reduce egg counts. stocking rate. Faecal egg counts will vary according to the parasite species involved and whether the worm burden consists of sexually mature parasites. Blood should be submitted to the laboratory chilled in insulated containers. v. ii. EDTA blood may not be suitable for differential white cell count and morphology if blood films are not made within 1 hour of blood collection. Leucocytes degenerate when blood stands for a few hours.HAEMATOLOGY BLOOD For red and white cell counts. Use additional WormTest kits for extra drench groups. STORAGE AND DESPATCH OF SPECIMENS Blood should be kept chilled. swampy areas. 20 ml. Use dry slides. BLOOD FILMS Fresh blood films should be made from EDTA blood immediately after collection. Avoid contamination when preparing blood films. iii. Draw it back until it touches the drop and the blood spreads evenly across the slide almost to the edges. PARASITOLOGY Faecal egg counts and egg type (by larval differentiation) are a guide to the size and type of worm burden.

Wormtest Kit samples should be collected from 10 animals. A serological test is used to show the presence or absence of antibody to a specific aetiological agent or group of agents. hair or wool must be clipped as close to the skin as possible. The jar should be filled and the lid closed tightly in order to minimise exposure of worm eggs to air which would allow egg development.Vacuum blood tubes (plain) For serology of liver fluke infection in cattle. Always include the head of the tapeworm. Faecal samples should be submitted as soon after collection as possible. i. wide mouthed bottle and submitted unpreserved. Gastrointestinal tract washings should be preserved in 5% formalin. protozoan and metazoan diseases. chilled in an insulated container. For tubes with anticoagulant ensure blood is gently mixed with anticoagulant. Parasites for identification Flukes. Mites Acute skin lesions should be scraped with a scalpel until blood is produced. rickettsial. When identification of parasitic cysts is required. Complete instructions for collection of samples are provided with each WormTest kit. The VETERINARY LABORATORY MANUAL 19 . double bagged in strong clear plastic bags. including apparently healthy and affected animals in each mob. COLLECTION OF SPECIMENS Faecal samples For individual animals at least 30 g of fresh faeces should be collected (preferably from the rectum) and placed directly into a jar. Menangle. Also see specific parasitic diseases: • Anthelmintic resistance in sheep • Babesiosis • Coccidiosis • Cryptosporidiosis • Cysticercosis • Drenching mortalities • Equine babesiosis • Fasciolosis in sheep and cattle • Ostertagiosis • Paramphistomiasis • Parasites internal/external/resistance • Sarcosporidiosis • Tapeworms • Tick fever • Toxoplasmosis • Trichomoniasis of cattle • Total worm counts (interpretation) • Worm egg counts (interpretation) SEROLOGY Serology is available for a range of bacterial and viral diseases. STORAGE AND DESPATCH OF SPECIMENS Faecal samples should be kept cool. Before scraping. between the stomach (abomasum) and small intestines and between the small and large intestines. Scrapings should be sealed in a small.48 hours.e. Only submit unpreserved chilled gastrointestinal tracts if delivery to the laboratory is assured within 36. tapeworms and roundworms should be washed in water and preserved in 5 per cent formalin. but not frozen. Scrape particularly at the edge of any visible lesion. Plain blood tubes should be allowed to clot at room temperature. Moisten the scalpel with liquid paraffin. Contact the laboratory for instructions on the procedure. tissues should be submitted chilled in a jar. Alternatively the organs can be washed by the field veterinarian and aliquots (from washings of known volume) submitted to the laboratory. Insects and snails for identification These should be submitted in 70% alcohol in a small leak proof container. Blood Collect a full tube of blood. mycoplasmal. Such tests may be available in Regional Veterinary Laboratories. Gastrointestinal tract The total gastrointestinal tract should be submitted direct to the laboratory when it can be delivered by the submitter. or only in Central Veterinary Laboratories at EMAI. Even prolonged chilling at 5oC kills the eggs of most species and makes samples unsuitable for larval culture for strongyle egg identification. The tract should not be opened and each part should be tied off at the appropriate junction. with icebricks wrapped in newspaper to prevent freezing. as well as for some chlamydial.

At least 5 ml of blood should be collected. A single serum sample is particularly useful in eliminating a diagnostic possibility. • Heating of samples.presence of antibody indicates exposure to the organism. etc. Since many animals in endemic areas may have antibody to a given organism. without anticoagulant. Use a separate sterile needle to avoid mechanically transmitting infectious agents from one animal to another. the best evidence of infection is the demonstration of a four-fold titre rise between samples collected early in the clinical episode and those collected 2-3 weeks later. COLLECTION OF SPECIMENS Blood samples should be collected aseptically. particularly in sheep and goat CFT's.. Frequently. • Contamination of the sample by water. • A slow flow from the needle. which is removed during testing. VETERINARY LABORATORY MANUAL 20 . provided there are no blood cells present in it. • Forcibly expelling blood through a needle. to reduce the risk of contamination of laboratory staff handling the specimens. • Freezing. as this leads to confusion and errors in reading numbers in the laboratory. Pig blood haemolyses quickly. Haemolysed or contaminated samples often give unreliable results in a complement fixation test (CFT): the serum may be anti complementary or give non-specific low titre positives. presence of antibody in the acute phase or absence in the convalescent phase will eliminate a diagnostic possibility. 10 ml These should be silicone coated. Do not label the stopper. or failure to insert into mid-vein. or after prolonged exposure to direct sunlight during collection.e. NB. Blood and faecal material should be removed prior to despatch. necessitating retesting of the animals involved (see Avoiding haemolysis of blood samples). preferably on an adhesive label. 5 ml For serum samples. The specimen advice form submitted with the samples should list clinical details beside each sample number. Serum can be frozen. A titre variation of less than four-fold (i. a single sample from an affected animal may not allow the serological test to be interpreted. using a 10 ml blood vacuum tube. It smudges when wet and may rub off if samples are chilled or frozen.g. usually in car boots or through back windows of car. Keep a key list which correlates sample numbers with animal identification. Labelling of samples Samples must be labelled serially (e. Serum should always be separated from the clot within 4 hours of collection. contaminated equipment or poor handling of the sample once it is collected. When a range of serological tests is required (particularly when viral serology and non-viral serology is to be undertaken). In tests where results are expressed in titres. STANDARD CONTAINERS AND EQUIPMENT Blood vacuum tubes. Poor quality samples will give poor quality results. DO NOT label containers with water soluble ink. one dilution) is within the normal variation of a serological test and is not significant. This will allow the laboratory to offer an informed comment on the results and perhaps apply other relevant tests. from 1 to 30) with a water proof pen. names. It also makes it very difficult for the laboratory to ensure all sample are present or to check on missing or broken samples. which may be due to a current clinical condition or to an earlier unrelated infection. Sterile screw capped containers. duplicate samples should be collected. Contamination of the container and stopper should be avoided. Avoiding haemolysis of samples Haemolysis occurs as a result of poor collection techniques. due to obstruction of the needle. Common causes of haemolysis include: • Use of non sterile containers for collection or storage. DO NOT label samples with tag numbers. • Contamination by faecal and other material due to faulty aseptic techniques.

Absorbance (Optical Density. Samples should be held in a warm room until the clot retracts. A GDPT reaction reflects the position of a specific line of reaction relevant to the serum and antigen wells. haemolysis and autolysis. complement and antigen are incubated then sensitised erythrocytes are added. OD) of the test serum. The level of ELISA-reactive antibodies can be measured in a qualitative and quantitative way. This colour is measured quantitatively and is termed the serum absorbance or optical density. ii. • a mathematical formula which expresses the OD of the test sample as a percentage of the positive control. where clearing of the tube due to agglutination is used to determine an endpoint. A reading of 2 indicates a line midway between the serum and antibody wells. whereas a 3 reaction is located closer to the antigen well. If there is specific antibody present. Results are given in titre form. with subtraction of the negative control OD from each.neg OD) / (pos OD neg OD) x 100 (Agar Gel Immunodiffusion (AGID) Syn: Gel Diffusion Precipitin Test (GDPT) Specific antibody is detected by placing test serums and control serum in wells in a gel around a central antigen well. or. Serum Agglutination Test (SAT) Tests for agglutinating antibody in a tubebased test. The second antibody is usually speciesspecific. 1. An absorbed ELISA (e. and the amount of colour which develops reflects the level of original serum antibody. blood samples must be held chilled to reduce contamination. Migration of antigen and antibody towards each other results in a visible line of precipitation where an antigen/antibody complex is formed. If virological examination of the clot is also needed (e. ELISA value or ELISA units depending on the test. iii. and the dilution series used (and therefore the titre) reflects the standard protocol for the test required.compares the OD of the test serum to that of a negative control. 2 or 3. originating from a range of positive and negative controls. This enzyme catalyses a colour reaction.STORAGE AND DESPATCH OF SPECIMENS Samples should be allowed to clot before transporting them over any distance. the endpoint readings are converted to international units rather than titres. Results are expressed as a titre. screw capped plastic container and submit the serum sample only. Clots may not retract readily in cold weather or if they are chilled too soon after collection. the test uses a second antibody system (directed against antibodies of the animal species under test) to which is linked an enzyme.g Pestivirus antigen detection). Test serum. ELISA ratio . but taking into account the performance of one or more positive control sera on the plate to give a valid test. If the laboratory will not get the samples within 48 hrs of collection. and 3 reflects the highest level of antibody detected. Enzyme-Linked Immunosorbent Assay (ELISA) The ELISA detects specific antibodies in serum which are allowed to bind to antigen on a solid phase. TYPES OF SEROLOGICAL TESTS Complement Fixation Test (CFT) The test quantifies complement-fixing antibodies to a range of antigens. A trace and 1 reaction are located closer to the serum well. ie: (test OD . reflecting a doubling serial dilution of serum from 1:4 upwards. this can be used to reflect either: • the OD of the test sample relevant to a standard curve. or. To detect the bound serum antibodies. decant the serum into a 5 ml sterile. The titre indicates the dilution at which 50% or more of the erythrocytes are not lysed. ELISA ratios of 2 or 3 are common cut-off points. an antigen-antibody complex is formed which binds complement and sensitised erythrocytes are not lysed. Thus each ELISA is geared towards one test animal species. submit clot separately. or. but also includes treatment of serum to absorb non-specific antibodies before testing. This may be in terms of: i. VETERINARY LABORATORY MANUAL 21 . Once the clot has retracted. In some tests. on a limited scale of trace.g. Trace is the weakest reading. Depending on the test. The quantitative measure is relative to control sera. Johne's ELISA) involves the above steps. but may react with closely related animal species at a different concentration.

Microscopic Agglutination Test (MAT) A test used for Leptospiral antibody which measures the ability of sera at varying dilutions to give an endpoint of 50% agglutination of one of a range of live leptospiral serovars. Latex Agglutination Test (LAT) Antigen coated-latex beads are reacted with diluted sera. Gives a qualitative (+/-) result only. Rapid Plate Test (RPT) An agglutination assay used for some poultry pathogens. Rose Bengal Test (RBT) A spot agglutination test performed on a solid base and used as a screening test for Br. and performed with diluted serum on a solid base. VETERINARY LABORATORY MANUAL 22 . Indirect Fluorescent Antibody Test (IFAT) A slide-based assay using serum at varying dilutions to show specific binding to the test antigen on the slide. abortus. The fluorescentlabelled second antibody provides a measurable signal of bound antibody from the test serum. with a positive test giving a lattice of latex beads due to agglutination.

CFT* M paratuberculosis Salmonellosis SAT (serogroup specific) S Typhimurium S Dublin Chlamydiosis/SBE CFT C psittaci Brucellosis ELISA. canicola. SAT* B abortus Liver fluke ELISA F hepatica Q fever CFT* C burnetti Neosporosis ELISA Neospora caninum Sheep Brucellosis CFT. Complement fixation test Enzyme linked immunosorbent assay Indirect fluorescent antibody test. australis. tarassovi. L tarassovi. L bratislava Brucellosis RBT. SAT B suis Poultry Pullorum RPT S pullorum Mycoplasmosis RPT M gallinarum M synoviae Dogs Leptospirosis MAT (serovar specific) L canicola L copenhageni Key AGID CFT ELISA IFAT LAT MAT RPT RBT SAT * Agar gel immunodiffusion test. CFT* M paratuberculosis Toxoplasmosis LAT T gondii Brucellosis SAT* B abortus Q fever CFT* C burnetii Horses Babesiosis IFAT* B equi Leptospirosis MAT*(serovar specific) L pomona L tarassovi Brucellosis/ SAT* B abortus Fistulous withers Pigs Mycoplasmosis ELISA M hyopneumoniae M hyorhinis Leptospirosis MAT (serovar specific) L pomona. zanoni Campylobacteriosis Campylobacter fetus venerealis ELISA (vaginal mucus) Johne's disease ELISA. seminis CFT (at ARI Yeerongpilly) A seminis epididymitis Johne's disease AGID. Latex agglutination test Microscopic agglutination test Rapid plate test Rose Bengal test Serum agglutination test.SUMMARY OF AVAILABLE SEROLOGICAL TESTS FOR VARIOUS NON-VIRAL DISEASES Host Disease Agent Available Diagnostic Test Cattle Leptospirosis MAT (serovar specific) L pomona L hardjo MAT* (serovar specific) Other Lepto serovars include: copenhageni. AGID. grippotyphosa. RBT*. Commercial test only VETERINARY LABORATORY MANUAL 23 . ELISA B ovis Salmonellosis SAT (serogroup specific) S Typhimurium S Dublin A. CFT* M paratuberculosis Chlamydiosis CFT C psittaci Liver fluke ELISA F hepatica Toxoplasmosis LAT T gondii Goats Johne's disease ELISA.

TOXICOLOGY
It is not possible to comprehensively screen samples for 'poisons' or 'toxins'. It is up to the submitter to consider the history, clinical signs and lesions (if any) and identify specific toxins for analysis. Contact your Regional Veterinary Laboratory if you are unsure whether a test is available for a particular toxin. In cases of suspected poisoning, it is important that an effect be demonstrated in the animal. For example, nitrate poisoning is confirmed by demonstrating the presence of nitrate in the serum or blood of the animal, not by demonstrating the presence of nitrate in pasture plants in the paddock. It is important that the history provided includes details of treatment with any suspected toxic compound, particularly in relation to the strength of the preparation and the time since treatment or access to the material.

Separate organs should be placed in separate containers. Body fat is the preferred tissue for insecticide residue testing. For biopsy material, a minimum of 2g is required. Blood samples At least 8 ml of blood in a blood vacuum tube, free from contamination with faeces etc. Serum samples At least 2 ml of serum should be submitted. Blood smears Thick air dried smears should be prepared, taking care to leave one end of the slide clean. They should be dry before being wrapped in paper. Suspected toxic material Suspected material, feedstuffs or plants should NOT be sent unless appropriate specimens from affected animals have also been sent. At least 50 g of material should be forwarded. Plant material Examination for nitrate nitrite and cyanide are best performed in the field. Plants for identification should be pressed and dried. Refer section on "Poisoning plant" for submission of plants for identification. Ingesta Nitrate and nitrite disappears rapidly from ingesta, and thus ingesta is of no value in diagnosing nitrate/nitrite poisoning. For other chemical poisons, eg. arsenic, lead, at least 250 g of ingesta should be submitted in an air tight, leakproof container.

STANDARD CONTAINERS AND EQUIPMENT
Blood vacuum tubes, 10 ml For blood samples Leakproof 5 ml tubes or bottles For serum samples. Bottles and jars For tissue samples Slides These must be clean. Some brands available have not been washed and are not suitable unless they are washed.

COLLECTION OF SPECIMENS
Particular care should be taken in collecting and packaging specimens for toxicology, because there is often a possibility that infectious agents are involved and these can create hazards to staff handling the material in a laboratory. Therefore avoid contaminating the outside of any submitted containers with tissues and ensure they are leakproof. Tissue samples At least 100 g of tissue should be collected, taking care to avoid contamination with soil, faeces or intestinal contents.

STORAGE OF SPECIMENS PRIOR TO DESPATCH
Tissues should be frozen. Herbage and ingesta samples for toxin examination should also be frozen. Other animal specimens should be chilled.

VETERINARY LABORATORY MANUAL

24

INTERPRETATION OF ARSENIC, LEAD AND COPPER CONCENTRATIONS IN CATTLE AND SHEEP Analyte Sample Units Deficient Normal Toxic Arsenic Liver mg/kg (wet wt) < 0.5 *>8 Lead EDTA blood µmol/L < 1.2 > 1.2 Kidney mg/kg (wet wt) <4 > 25 Faeces mg/kg (wet wt) < 10 > 25 Copper Liver mg/kg (wet wt) <4 ** 20-70 † > 100 Kidney mg/kg(wet wt) 4-6 >8 * ** † NB i. ii. iii. • • • Liver arsenic may be in range 2 to 8 mg/kg if several days elapsed since toxic exposure. Typical liver copper for cattle is > 20 and sheep > 40. In sheep, liver copper may increase up to 200 mg/kg (wet wt) before poisoning occurs

Interpret concentrations between normal and toxic according to clinical and pathological findings. Concentrations based on dry wt are approximately 5 times the above (wet wt) values. Conversion from mg/kg to SI units is as follows: As mg/kg x13.3 = As μmol/kg Pb mg/kg x 4.8 = Pb μmol/kg Cu mg/kg x 0.0157 = Cu mmol/kg

VIROLOGY
Virological examinations involve demonstration of a pathogenic virus or detection of antibody to virus. Findings must be interpreted in the light of history, clinical findings, lesions, etc. It is not possible to screen for a wide range of viruses. Submitters should forward specimens to be tested for specific viruses. If there is any doubt about the availability of a test, the laboratory should be contacted for advice.

Presence of antibody can be a result of clinical disease, unapparent infection, passive immunity or vaccination. Tests variously demonstrate group specific antibody, type specific antibody or a cross reaction. Viral serology can be applied with the following limitations: Single Serum • Useful only to eliminate a diagnostic possibility. Presence of antibody in the acute phase may eliminate the diagnostic possibility; absence of antibody in the convalescent phase eliminates the diagnostic possibility. • Accurate interpretation often difficult because time of collection may be critical. A negative test on serum < 3 weeks after a suspected viral condition leaves the possibility that the animal was infected but had not yet produced detectable antibody. Paired Sera • A change from negative to positive antibody status is known as seroconversion and indicates infection. • A four-fold rise in antibody titre between acute and convalescent samples can also indicate infection by the specific pathogen. However both the above may also be due to cross reaction or booster immunisation. They may also indicate stress induced reactivation of latent infection.
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DIAGNOSIS OF VIRAL DISEASE
The diagnosis of a viral disease can be based on Histopathology, Virus isolation, Virus Serology, or Virus or Virus Antigen Detection. HISTOPATHOLOGY Cytological changes can indicate a viral aetiology. VIRUS ISOLATION Cultivation and identification of virus grown in tissue culture or eggs inoculated with specimen. NB Failure to cultivate the virus does not rule out a viral aetiology. Cultivation of a virus does not necessarily mean it caused the disease process. VIRAL SEROLOGY Detection and quantitation of virus specific antibody in the serum of an infected animal. Plasma is also an acceptable sample for most serological tests.
VETERINARY LABORATORY MANUAL

The relationship between virus and antibody varies between diseases. In some, virus does not occur in animals with detectable antibody, e.g., Akabane, Ephemeral Fever whereas in others, virus and antibody can be present in the one animal, e.g., EIA, IBR, CAE, EBL and occasionally Pestivirus. NSW Department of Primary Industries offers the following types of viral serology tests: • Agar Gel Immunodiffusion Test (AGID) • Virus Neutralisation Test (VNT) • Haemagglutination Inhibition (HI) Test • Enzyme Linked Immunosorbent Assay (ELISA) Agar Gel Immunodiffusion (AGID) Test AGID involves diffusion of viral antigen and antibody towards each other through a gel. When they combine, they precipitate in the gel. This produces a visible line where the concentrations of antigens and antibody are balanced. An excess of antigen or antibody can alter the location and appearance of the precipitin line. Each test sample for viral antibody is tested against a known viral antigen to examine the relationship of any precipitin line formed with an adjacent standard reference line formed between known positive serum and virus antigen. This reference line is optimised to give a strong visible precipitin line centrally located between the antigen and positive serum wells (termed a 3 reaction - see below). For viral serology, a specific precipitin line formed by the tested serum sample is recorded as 1, 2, 3, or >3 to describe its relative position to the serum well and the antigen well: Description / Position of Precipitin Line Turn on end of reference line Line, closer to serum well Line, midway between serum well and antigen well Line, closer to antigen well A 1, 2, 3 or >3 antibody reaction is a positive test for antibody, and indicates that an animal has been infected with the specific virus (or a related virus). Strength of viral antibody levels in the test generally does not reflect the severity and stage of infection with any certainty. Virus Neutralisation Test (VNT)

Serum is usually titrated in two fold serial dilutions beginning at 1/4 or 1/10. The virus neutralising antibody titre is the reciprocal of the serum dilution that will neutralise a standard amount of virus (usually 100TCID50). Two serial dilutions rise (i.e. 4-fold) in titre in the convalescent serum sample compared to the acute serum sample is considered significant. This occasionally occurs as a result of technical or statistical variation, so greater differences can be accepted with greater confidence. Haemagglutination Inhibition (HI) Test HI tests are used to detect antibodies to viruses which agglutinate erythrocytes. HI tests depend on antibody binding a fixed amount of antigen, preventing it from causing haemagglutination. Sera is tested in two-fold dilutions, and results expressed as a titre (negative, 2, 4, 8, .......) etc. More accurate measurement of flock status can be gained by repeat sampling of the flock to determine if a rise in titre (e.g. a rise in titre of two dilutions or more is arbitrarily considered significant) has occurred. Enzyme linked immunosorbent assay (ELISA) ELISA results can express the detection of antibody in a qualitative (+ or -) or a quantitative way. The three common means of expressing a quantitative result are absorbance (optical density), ELISA ratio and ELISA value (unit). Absorbance (Optical Density) The amount of antibody in a sample is proportional to the absorbance (optical density, OD) given by it. The OD may be standardised against one or more serum controls on the ELISA plate. ELISA ratio The ELISA ratio indicates the strength of the sample OD compared to that of a Result negative 1 serum control on the same ELISA plate. (E/R = OD test/OD neg 2 control) 3 ELISA value or ELISA units >3 The sample OD is fitted to a curve determined by the performance of control positive and negative samples, and the result given as an ELISA value (eg 0 100). This is often a measurement of how the sample OD compares with that of a high positive control (taken as a value of 100). Percent inhibition The results of blocking or competitive ELISAs are expressed as the reduction in OD (as a %) relative to the OD given
26

VETERINARY LABORATORY MANUAL

Rotavirus. Plain sterile swabs are used and immediately placed in small bottles of phosphate buffered gelatin saline (PBGS). 2. Please do not use serum separator tubes. eg: • Newcastle Disease • Avian Influenza • Egg Drop Syndrome for On poultry tissue or allantoic fluid from eggs.by a reference positive monoclonal) antibody. or XS) against the reference line. VIRUS OR VIRUS ANTIGEN DETECTION The following types of test are available for the direct detection of virus or viral antigen: • Electron microscopy (EM) • Agar Gel Immunodiffusion (AGID) Test • Enzyme linked Immunosorbent Assay (ELISA) • Latex Agglutination Test • Haemagglutination (HA) Test Electron microscopy (EM) Allows detection and identification of virus particles on a morphological basis e. ratios or values may be placed into categories (negative/inconclusive/positive) determined by previous experience of the performance of the test. COLLECTION OF SPECIMENS Specimens for virus isolation must be collected by aseptic techniques. The containers and equipment listed are suitable for the routine collection of tissues for virus examination. or detection of antigen to. (often ELISA ODs. Results can be given as a titre derived from a doubling dilution of the sample (e. 4. originating from an adjacent positive antigen. To determine which virus is responsible. Swabs and PBGS bottles: For discharges. STANDARD CONTAINERS AND EQUIPMENT Blood vacuum tubes. The results are qualitative (+ or . Latex Agglutination Test Can be used to detect viral antigen or virus particles. which is continuous through a turn of identity with the reference line is classed as +++ for viral antigen. HA positive samples can be tested in the presence of specific antibody in a direct (or reverse) HI test. etc. A test sample giving a line of similar position and intensity. organs Sterile 5 ml vials: For serum and fluid samples. the sample is tested against a known positive serum and the relationship of any precipitin line formed with a standard reference line. 8. +++. Poxvirus.etc). In such cases.). using VETERINARY LABORATORY MANUAL 27 . Sterile instruments An adequate range of sterile scissors and tissue forceps should be available.. Containers and equipment for special requirements will normally be supplied by the laboratory. ++. These are available from the Regional Veterinary Laboratories.g. Agar Gel Immunodiffusion (AGID) Test Antigen detection is useful where virus infected tissues contain soluble virus antigens that visibly precipitate with specific antibody. eg: • Pestivirus detection in the Pestivirus antigen capture ELISA (PACE). eg: • Parvovirus in mummified pig foetuses • Pestivirus in gut scrapings of cases of mucosal disease • Rotavirus in faeces.. Results can be expressed qualitatively (+ or ) or quantitatively (OD's or ELISA ratios or ELISA values). some viruses). is examined. Sterile bottles for samples of tissues. The results are recorded in terms of relative strength (+. after prior arrangements have been made. eg: • Rotavirus detection Haemagglutination (HA) Test Can be used to screen haemagglutinating viruses. silicone coated For serum samples and blood clots (for isolation of. Enzyme linked Immunosorbent Assay (ELISA) Can be used to detect either virus antigen or intact virus particles. Results are only qualitative (+ or ). Blood vacuum tubes with heparin or EDTA For blood for virus isolation (of mainly the arboviruses) or preparation of buffy coat samples for antigen detection.g. .

Blood This should be collected using the appropriate blood vacuum tube. A separate needle should be used for each animal. (VI) Antigen ELISA. VI VNT. as this creates difficulties in handling the specimens within the laboratory. Avoid contamination of the outside of the tube by blood. Faeces At least 10 g of fresh faeces should be collected into a clean jar. avoiding contamination of the sample. Avoid contamination from other tissues as well as that from extraneous sources. mucosal surfaces and orifices should be taken SUMMARY OF AVAILABLE VIROLOGICAL TESTS FOR VARIOUS DISEASES OR DISEASE SYNDROMES Section A: Recognised Viral Diseases Host Disease Cattle Ephemeral fever (EF) Bovine pestivirus Mucosal disease (MD) Infertility Abortion Congenital abnormalities Bovine malignant catarrh Virus Rhabodovirus Pestivirus Available Diagnostic Test VNT. which may otherwise become frozen. specimens except blood samples should generally be frozen. Submissions for virus isolation should be accompanied by serum from the affected animal(s) whenever possible. Serum If blood samples cannot arrive at the laboratory within 2 days. especially after freezing. Swabs Swabs from excretions. avoiding contamination from other sites. STORAGE AND DESPATCH OF SPECIMENS All virological specimens should be chilled prior to and during transport. However. The clot should also be submitted as it may be required for virus isolation or antigen detection. Each swab should then be transferred to PBGS at room temperature. then chilled (but never frozen) for transport to the laboratory. VI VNT. care should be taken to prevent direct contact between coolant bricks and specimens. VI Antibody ELISA. Scabs Scabs and underlying tissue should be submitted in a sterile bottle. use histopath PCR at AgWest VNT.g. carefully. Check that screw caps are tight. All samples should be packed in insulated containers with sufficient icebricks to ensure that they are still cold when received at the laboratory. Tissues and organs These must be collected aseptically. If more than 48 hours is to elapse between collection and receipt at the laboratory. IBR). (ELISA). These viruses have extremely poor survival at o 20 C with just a single freeze. Portions of tissue (no greater than 2 cm x 2 cm x 2 cm) should be taken. the serum should be poured off aseptically into a sterile 5 ml vial. and tissues for their isolation must be kept chilled. For isolation of Arboviruses (especially Bluetongue) and Herpesviruses (e. Contamination between tissues must also be avoided. using separate sterile instruments and containers for each tissue or organ. All specimens should be clearly labelled and sent in a leakproof container. VI VETERINARY LABORATORY MANUAL 28 .sterile instruments and sterile containers. exudates. specimens should never be frozen. AGID. soil and faeces. VNT Infectious bovine rhinotracheitis (IBR) Infectious pustular vulvovaginitis (IPV) Encephalitis Bovine herpesvirus Ovine herpesvirus 2 (OHV2) Bovine herpesvirus – Type 1 Bovine herpesvirus – Type 1 Bovine herpesvirus – Type 1 Nil.

VI) EM. Antibody ELISA. Histopath AGID. AGID. Histopath Histopath HI. Histopath AGID. VI) AGID. VNT. EM AGID (for antigen and antibody) ELISA. ELISA. VI) AGID. HA. ELISA. VNT is type specific AGID. HA. VI. VI) AGID. Histopath AGID is group specific. AGID) VI. ELISA Antigen ELISA. Histopath (HI) Bluetongue virus (BTV) (various types) Pigs Encephalomyocarditis virus (EMC) Swine Pox Haemagglutinating encephalitis virus infection (HEV) (Vomiting and wasting disease) Piglet scours Parvovirus disease Cardiovirus Poxvirus Coronavirus Rotavirus Parvovirus Goats Horses Poultry Caprine arthritis encephalitis (CAE. AGID is group specific. (VI) Abortion congenital abnormalities Sheep Ibaraki disease Contagious pustular dermatitis (Orf. (VNT) EM. (VNT. VI. (VNT) AGID VI. VNT AGID. EM EM ELISA. (VNT. scabby mouth) Arthrogryposis/hydranencephaly (AG/HE) Ovine pestivirus (Border disease) Bluetongue Palyam virus (various types) (see also Pestivirus) Enzootic haemorrhagic disease of deer (EHD) virus (EHD1 EHD5) EHD2 Parapox virus Akabane virus Pestivirus LA. AGID is group specific. VI NB. VNT is type specific VI. VNT is serotype specific 29 Retrovirus Haemagglutinating adenovirus 127 (group II) Adenovirus (group I) (1 serotype of 12) VETERINARY LABORATORY MANUAL . VI NB. (VNT) ELISA. ELISA. (VI. (VNT. VNT. VNT. AGID AGID. VNT. (VNT. VNT. big knees) Vaginitis Equine infectious anaemia (EIA) Equine rhinopneumonitis Viral abortion Avian encephalomyelitis (AE) Avian influenza Fowl plague/ Duck influenza Avian leukosis Egg drop syndrome (EDS 76) Fowl adenovirus (FAV) acute inclusion body hepatitis (IBH) of chickens < 14 d Retrovirus Caprine herpesvirus Retrovirus Equine herpesvirus –Type 1 Picornavirus Orthomyxovirus LA.Host Disease Mammilitis Bovine papular stomatitis (pseudocowpox) Congenital arthrogryposis/hydranencephaly (AG/HE) Calf scours Enzootic bovine leucosis (EBL) Bluetongue Virus Bovine herpesvirus – Type 2 Parapox virus Akabane virus Aino virus Palyam virus Pestivirus also causes AG Rotavirus Coronavirus EBL virus (Retrovirus) Bluetongue virus (BTV) (various types) Available Diagnostic Test EM EM AGID.

Histopath Herpesvirus Adenovirus (group I) (Many serotypes) Coronavirus Birnavirus Herpesvirus Herpesvirus Paramyxovirus Retrovirus Iridovirus (VI).Host Disease Fowl Pox (FP) Turkey haemorrhagic enteritis (THE) Haemorrhagic enteritis (HE) of turkeys Herpesvirus of turkeys Inclusion body hepatitis (IBH) Infectious bronchitis (IB) Infectious bursal disease (IBD) Infectious laryngotracheitis (ILT) Marek's disease Newcastle disease (ND) Reticuloendotheliosis (RE) Epizootic haematopoietic necrosis (EHN) White spot syndrome (WSS) Virus Avipoxvirus Adenovirus (group III) Available Diagnostic Test EM. Histopath (VI). Histopath. Histopath Antigen ELISA. Histopath VI Fish Crustacea White spot syndrome virus PCR Section B: Syndromes with Possible Viral Involvement Host Cattle Disease Respiratory disease Possible Virus Parainfluenza 3 (PI3) Respiratory syncytial (RSV) Pestivirus Rhinovirus Adenovirus PI3 ? Available Diagnostic Test ELISA. (VI). ELISA. VNT) * AGID ELISA. (VNT) ELISA. (ELISA. HA. (VI). (AGID). Histopath ELISA. (VNT) AGID. Histopath Sheep Poultry Respiratory disease Big liver spleen syndrome (BLS) Key AGID ELISA EM HA HI LA VI VNT * Agar gel immunodiffusion test Enzyme linked immunosorbent assay Electron microscopy Haemagglutination test Haemagglutination inhibition test Latex agglutination test Virus isolation Virus neutralisation test available by arrangement VETERINARY LABORATORY MANUAL 30 . VI. (VI) (AGID. Histopath HI. VI. VI). Histopath AGID. Histopath VNT. (HVT) VI. (VNT) AGID. VNT.

and. Sections of cotyledons. not frozen. Akabane virus and pasteurellosis. placenta. or ii. virology. and a range of tissues and body fluids submitted to eliminate cause of perinatal death other than infectious agents e. Pestivirus. akabane and palyam are predominantly associated with abortions of the last trimester of pregnancy. has recovered from tissues of affected foetuses. listeriosis. brucellosis. In some regions (e. ABORTION IN SHEEP AND GOATS Common causes in New South Wales include toxoplasmosis. Indicate on the Specimen Submission Form which animals have aborted. hypocalcaemia. pregnancy toxaemia. brain. liver. lung. Portions of foetal stomach contents. salmonellosis. This test is available for cattle and pigs. Where it is not feasible to submit the foetus. and kidney in buffered formalin for histopathology. myocardium. Infectious abortions can be distinguished from non-infectious abortion by measuring elevated levels of species-specific IgG in foetal fluid (blood. brain. lung. liver. Illawarra and South Coast). there is little value in laboratory investigation. septicaemia. Diagnosis Based on bacteriology.g. iv. peritoneal. Clear serous body fluid (pericardial. Habitual abortion can occur in Angora does. a sporozoan with a canid-bovine transmission cycle. Sera from the aborting animals and 10-15 animals from the same group. They should be chilled. campylobacteriosis.g. fungal and viral infections of the dam and foetus. The whole foetus and foetal membranes submitted chilled. Indicate on the Specimen 31 ABORTION IN CATTLE Common infectious causes of abortion in New South Wales are leptospirosis. iii. liver. the aborted foetus together with the foetal membranes should be submitted to the laboratory. serology and histopathology findings together with the previous clinical history of the dam. Foetus and foetal membranes submitted whole in an insulated container. Other causes include chlamydiosis and conditions causing clinical illness in the ewe or doe. and. thoracic or peritoneal) or heart blood submitted chilled in separate sterile containers. Portions of foetal stomach contents. or: ii. Akabane and Palyam virus possibly cause abortion. the most commonly diagnosed infectious cause of abortion is neosporosis. Specimens required Aborted foetuses i. thoracic or peritoneal) or heart blood submitted chilled in separate sterile containers. myocardium. Ewes and does i. and dystocia is indicated.SPECIMENS (BY DISEASE OR SYNDROME) DISEASES OF LIVESTOCK ABORTION (GENERAL) Notes on the individual species should be consulted. spleen. Whenever possible. Non-infectious causes include plant poisonings and congenital abnormalities. dystocia. and campylobacteriosis. liver. Clear serous body fluid (pericardial. lung. Sera from the aborting animals and up to 10 flock mates submitted chilled. Pestivirus may be associated with abortion at any stage of gestation. the autopsy must be complete. pleural or pericardial fluid). iv. placenta. Every opportunity should be taken to emphasise to owners the importance of collecting the foetus and foetal membranes.g. Neospora caninum. Sections of cotyledons. lung. If a full term calf is examined in the field. These three viral infections may cause congenital abnormalities. e. North Coast. Border disease. spleen iii. Cows i. Specimens required Aborted foetuses i. VETERINARY LABORATORY MANUAL . and kidney in buffered formalin for histopathology. Characteristic lesions are seen in sections of fixed brain and myocardium. Other infectious causes include listeriosis.

epididymes and accessory sex glands including ampullae and seminal vesicles submitted chilled. Escherichia coli. Testes. Syn: Ketosis See also: Pregnancy toxaemia in sheep Occurs in both beef and dairy cattle Diagnosis History. NB Parvovirus infection causes foetal mummification and parturient deaths. ii. Serum sample submitted chilled (for beta hydroxybutyrate analysis).. ACTINOBACILLOSIS AND ACTINOMYCOSIS Diagnosis Demonstration of Actinomyces or Actinobacillus organisms in pus or tissue sections from early lesions. Demonstration of A seminis on culture of semen. brain and spleen submitted in buffered formalin for histopathology. Serum sample. clinical signs.. Specimens required i. ACTINOBACILLUS SEMINIS INFECTION IN RAMS Diagnosis History. Affected tissue and specimens of pus submitted chilled. kidney and spleen together with foetal stomach contents submitted chilled. lung. typical histological changes. serum biochemistry (beta hydroxy butyrate). iii. ABORTION IN PIGS Common causes of abortion in swine in New South Wales include leptospirosis. oestrogenic feeds. particularly starvation and/or stress in late pregnancy and early lactation. Serum sample from the aborting sow and up to 10 herd mates. Other possible causes include genetic factors. or. Specimens required i. Aborted foetuses and membranes submitted chilled. Portions of placenta. lung. Portions of placenta. smothering and chilling. ii.. Klebsiella spp. kidney. ACETONAEMIA ABORTION IN HORSES In the past. Sections of recent lesions in buffered formalin for histopathology. At least four pus smears from deep parts of the lesion. Specimens required i. equine herpesvirus and fungi. Non-infectious causes may include maternal pyrexia and malnutrition.Submission Form which animals have aborted. Smears from the surface are not satisfactory. Specimens required i. Specimens required Aborted foetuses i. Granulomatous lesions containing granules in the pus may also be caused by staphylococci and Arcanobacter (syn Corynebacterium) pyogenes. adrenal gland. endocrine dysfunction. See Abortion in cattle re specimens for virology ii. Foetus and foetal membranes submitted whole and chilled in an insulated container. erysipelas.. leptospirosis. it has frequently been impossible to reach a diagnosis because of an inadequate history and unsuitable specimens submitted. Salmonella spp. submitted chilled (to eliminate Brucella ovis as a cause). encephalomyocarditis (EMC) virus and vitamin A deficiency. Staphylococcus spp. demonstration of ketones in urine and milk. liver. NB Examinations of urine and milk for ketones and glucose should be carried out in the field. submitted chilled. Abortion should be differentiated from parturient and immediate post parturient deaths due to hypoxia and managerial factors such as injury or stress. ii. cord compression abnormalities. iii. 32 VETERINARY LABORATORY MANUAL . liver. submitted chilled. endometrial incompetence and immunological factors. Semen sample collected aseptically. it does not normally cause abortion. streptococcal infection and toxaemic conditions in the sow. Common infectious agents responsible for equine abortion include Streptococcus spp. clinical examination (particularly for orchitis in young rams). iii.

skeletal and cardiac muscle and red bone marrow from the femur in buffered formalin. • Babesiosis. ii. • Parasitism. Serum chilled for copper etc. spinal cord and muscle in buffered formalin for histopathology. and aflatoxin analysis where appropriate. ALPHA MANNOSIDOSIS OF CATTLE See: Mannosidosis ANAPLASMOSIS See: Tick fever ANAEMIA This can arise from acute or chronic blood loss. appropriate tissues should be sent. kidney. The diagnosis of the cause requires the submission of the appropriate range of specimens. • Coccidiosis. Brain. which can cause arthrogryposis (AG) and CNS lesions. rape. Liver. fascioliasis and enzootic haematuria are best diagnosed in the field. Faeces for parasitology v. by increased erythrocyte destruction or by impaired erythrocyte production. Specimens appropriate to the particular clinical syndrome should be submitted. iii. or. pyrrolizidine alkaloids. The following disease conditions should be considered. pathological findings and the demonstration of antibodies to Akabane virus in foetal fluids or serum samples taken from calves or lambs which have not fed from the mother. AKABANE DISEASE IN SHEEP AND CATTLE See also: Congenital abnormalities. including haemonchosis. abortion. from a neonate. arthrogryposis and hydranencephaly' Diagnosis The diagnosis of Akabane disease relies on history. iv. Diagnosis Based on clinical findings. Many conditions. Irrespective of the syndrome. • Enzootic haematura in cattle. serum if the animal has not sucked. anaplasmosis and eperythrozoonosis. • Plant poisonings.NB The CF test on serum is not a reliable test for A. ALGAL POISONING See: Blue-green algal poisoning' AFLATOXICOSIS Aflatoxins can cause a range of clinical signs including nervous signs. submitted chilled. histopathology. including fascioliasis and other helminthiases. ii. including bracken fern. Serum for enzymology. iii. Specimens required i. serum enzymology. hepatic disorders and abortions. sarcosporidiosis. ANNUAL RYEGRASS TOXICOSIS (ART) Toxicity of Wimmera ryegrass to cattle and sheep in WA and SA is associated with a plant toxin of the tunicaminyluracil antibiotic complex produced in galls in the rye grass seed head by parasitic bacteria (Clavibacter formerly Corynebacterium rathayi) 33 VETERINARY LABORATORY MANUAL . Specimens required i. spleen. Diagnosis The diagnosis of clinical anaemia should be made in the field. ii. • Iron deficiency anaemia in piglets. Specimens required Depending on clinical and post mortem findings. • Copper and cobalt deficiency in ruminants. These should include: i. Blood films for haematology. seminis. At least 5 ml EDTA blood for haematology. Suspected foodstuff for aflatoxin analysis and mycology. NB Specimens should also be submitted to exclude pestivirus infection. At least 2 ml of foetal fluid (preferably pericardial or pleural rather than heart blood or peritoneal fluid which are more contaminated) for virus serology. (This is only useful in eliminating a diagnosis of Akabane or Aino infection) iii. diarrhoea. sections of liver and kidney in buffered formalin should be submitted to check for toxic changes. Serum sample from the dam.

the trial should contain groups treated with a range of anthelmintics. other drenches including naphthalophos (NAP. For more information see: http://www. or. However. originating from infected "blowaway grass" (Agrostis avenacea). DrenchRite™) is especially useful for initial drench resistance testing on a property as it requires little on-farm effort. histopathology. and moxidectin should be considered.au/reader/dasvettesting/2566 Diagnosis History. with recent extensive spread of resistance in these chemical groups. clinical findings. ending in death. Ultimately treatment becomes ineffective because a large proportion of the parasites in the population are resistant.g. levamisole (LEV).nsw. ii. respectively) should be included if possible. Evidence of yellow bacterial galls on annual ryegrass plant heads/seeds. Clinical signs of ART include neurological disturbances characterised by ataxia and collapse with tetanic and clonic convulsions. ie avermectin/milbemycins). Fixed liver and brain in buffered formalin ii. This disease has not yet been recorded in NSW. VETERINARY LABORATORY MANUAL . they are used to obtain a prompt answer to possibly a serious and immediate problem on specific farms.gov. it is emphasized that the use of double dose rate or combinations of anthelmintics are not general recommendations. or on-farm trial using a faecal egg count reduction test (FECRT). FECRT pre-trial check sampling (Day 10) 34 ANTHELMINTIC RESISTANCE Repeated treatment of populations of parasites with anthelmintics selects individuals that have innate or acquired resistance to the drugs. and other MLs such as abamectin. but a similar disease involving the same toxin and Clavibacter bacterium has been seen in cattle and some sheep in late 1990 along the Darling flood plains. Traditionally 5 groups (BZ. Rametin ™).g. FECRT can be performed using either: i. and its combinations with BZ and LEV. enterotoxaemia). Drench resistance is common and often highly developed in goats. FECRT anthelmintics Where nematode resistance is suspected.transported by invading plant nematodes (Anguina agrostis). A DrenchTest kit. Groups may be included where treatments are given at greater than the recommended dose rates to fully evaluate the available anthelmintics. combination (BZ/LEV) and indications for macrocyclic lactones (ML. failure to respond to treatment. closantel (CLS). BZ/LEVcombination. IN VITRO LARVAL DEVELOPMENT ASSAY (LDA) In vitro larval development assay (LDA. ivermectin (IVM) and an untreated control group) were used. Use of special formulations of triple or quadruple combinations (Triton™ and Q-drench™. in vitro larval development assay (LDA. and abortions in ewes. Several WormTest kits (one for each anthelmintic and one for a control group). However. It is becoming disturbingly common for worms to be resistant to anthelmintics in all of the major drench groups. It is now common for sheep to harbour at least one parasite species that is resistant to one of the major drench groups. Suspect grass or grass seed (e. showing bacterial galls). identification of Clavibacter sp in feed. In practice. Specimens required i. Since 2002 half-dose ivermectin and 1/3dose CLS have been recommended for the early detection of emerging resistance. Diagnosis History. gross pathology. Due to their altered drug metabolism goats may require higher dose rates than recommended for sheep. However. It can not be used for increased dose levels or combinations. it is only able to quantify resistance to benzimidazole (BZ). LEV. Only a limited range of anthelmintics are registered for use in goats. FAECAL EGG COUNT REDUCTION TEST (FECRT) Faecal egg count reduction test (FECRT) requires considerable on-farm effort. DrenchRite™).agric. Elimination of other diseases associated with neurological signs and CNS oedema (e. only double dose LEV or double dose LEV plus single BZ have been shown to be effective where the single dose is not effective.

Anthelmintic treatment of trial animals Anthelmintics should be administered accurately by either a calibrated syringe or a drench gun previously calibrated. the next 5 are similarly allocated until each group contains 15 sheep. Subjective evaluation of the data may however be possible below this figure. This is important as some species have different susceptibility to different drenches (eg Levamisole may still be effective as a narrow spectrum against Haemonchus. together with a 95% confidence interval. Randomisation of trial animals Sheep should be allocated to treatment and control groups randomly. if 4 drenches are to be tested.e. of even body weight and bred on the property. Number of trial animals Each group should contain 15 sheep allowing that faeces may not be collected from all animals at the subsequent visit. FECRT treatment (Day 0) Selection of trial animals Trial sheep should be preferably be young sheep at weaning. A minimum of 5 g (10 to 15 pellets) should be collected from each sheep. as these are usually not available. The best method of randomisation is based on pretreatment egg counts. but not as a broad spectrum against other species). If this is not possible. Some authorities even suggest 300 epg. The samples should be kept cool in an esky with freezer bricks wrapped in newspaper to avoid freezing. should be transported to the laboratory as soon as possible. most accurately describes the range of in efficacy of an anthelmintic. The arithmetic mean is the most stringent test of a percentage reduction in faecal egg output and therefore is a more conservative measure of an anthelmintic's performance. Calculation of FECR results The arithmetic mean (‘average’) on epg of samples collected 10-14 days after treatment. it is of value to arrange for a pre-trial collection of faeces to obtain an estimate of the egg count of the flock. contortus eggs in a faecal culture. If the group is an even line of sheep the dose received on a mg/kg basis will not vary greatly. FECRT sampling (Day 10-14) Collection of faeces VETERINARY LABORATORY MANUAL All trial sheep should be sampled between 10 and 14 days after treatment. the first 5 sheep in the race are allocated at random to 5 groups.To avoid wasted effort. but arithmetic calculations should always be conducted as well. = FECR) • 95% confidence interval of FECR (Upper and lower limit of percent reduction for 95% confidence interval) 35 . Each set of results will include the following information per group for each genus present (provided a larval differentiation has been conducted): • Arithmetic mean (‘Average’) • Range (Highest and lowest counts) • Standard deviation • Mean percent reduction in epg (i. This will allow determination of the species composition and species resistance levels. sheep should be orally dosed according to the weight of the heaviest animal in the group. faecal samples may be collected from the control group at day 0 to assess if egg content in faeces is sufficient to continue the trial. FECRT laboratory testing Larval differentiation Pooled faecal cultures and larval differentiations should always be carried out on the control group and preferably each of the test groups. Undrenched weaners are preferred. Transportation of faeces Samples in individually sealed containers filled as close to the top as possible to exclude air. In most situations. (A minimum of 10 sheep per group from which collections are made is necessary). Objective assessment of the efficacy of an anthelmintic is not reliable where the mean faecal egg count of the control group is less than 200 epg. Geometric means are sometimes requested by trial sponsors. For example. Storage or transport of faeces at or below 4oC may affect the hatching of H. Each group should be identified with a different colour marker (coloured ear tag is often the most convenient). Faecal samples should be collected from the rectum of sheep within the first hour after yarding and before much handling has occurred. 3-6 months of age. the sheep should be drafted into as many groups as there are anthelmintics to be tested plus one untreated control group. however. but if not possible they should NOT have received a drench during the previous 6 weeks or 8-12 weeks (for persistent ML anthelmintics) to avoid testing an already ‘selected’ worm population.

ANTHRAX Anthrax is a notifiable disease. due to the costs. Where anthrax is suspected. Blood smears from peripheral blood vessels. e. a positive Reinsch test (indicates >1mg arsenic/kg) on rumen contents from animals with a suggestive history and 36 . smears from any affected organs. Smears should be sent from more than one animal. ii. iii. i. *0 Levam 190 800 0 287 *0 69 *0 BZ+Levam 23 80 0 31 * 85 96 * 43 Comment: Interpret these results with caution due to low egg counts in the controls. Specimens required i. BZ + LEV resistance (*) is present. From animals other than sheep and cattle. NB Arsenic analysis of tissue is not routinely undertaken. VETERINARY LABORATORY MANUAL ii. • Make thick. ii. 82 Lower 95% conf. causing serious illness and sometimes death in humans. 250 g ruminal or stomach contents submitted chilled for toxicology (Reinsch test). An example of simple test groups without larval differentiation: Control BZ Arithmetic mean 155 76 Highest 520 160 Lowest 40 0 Std Dev 168 72 % Reduction * 51 Upper 95% conf. ii. Caution Anthrax is a zoonotic disease. However it appears that: i. swellings in throat and lymph nodes of pigs. LEV resistance (*) is present. the lower 95% confidence limit of the % reduction in egg count is less than or equal to 90%. clearly labelled and submitted chilled. Diagnosis Demonstration of Bacillus anthracis in blood and tissues. and.g. NB Always clearly label specimens as ‘Suspect Anthrax’ Pack securely and forward separately from other specimens Advise the laboratory that you are submitting the samples and place a warning under the lid of the outer packaging. BZ resistance(*) is present. leaving one end of the slide clean. Where anthrax is suspected as a result of changes seen at necropsy then impression smears of tissue should be submitted in addition to a range of fresh and fixed tissue to establish an alternative diagnosis if necessary. clinical and post mortem findings. detection of arsenic in milk or urine. These spores will remain a source of infection for long periods. the FECR is < 95%. Fees for tests undertaken to confirm or exclude a diagnosis of anthrax are paid by NSW Department of Primary Industries. Specimens required Each specimen should be packed in a separate container. a post mortem should not be undertaken as it will cause the vegetative forms of Bacillus anthracis to sporulate. A diagnosis of anthrax can be made from smears of peripheral blood obtained by cutting the ear of the unopened carcase. In most cases. • Do NOT fix smears by heat or other agents. with gastritis or enteritis. detection of a significant concentration of arsenic in the ingesta or tissues of the dead animal. In chronically exposed animals. submitted chilled for toxicology.Interpretation of FECRT results Resistance should be regarded as being present if: i. This will ensure special biosecurity precautions are undertaken at the laboratory ARSENIC POISONING Diagnosis History. 50 g of liver from the dead animal. air-dried blood smears. • Fix blood smears with methanol prior to submission.

Brain. muscular.5 mg/kg. circulatory or respiratory defect.25-1. Diagnosis of the cause depends on history and samples from the dam and affected animals. Sections of extensor muscles in buffered formalin for histopathology. and Pestivirus (Mucosal Border disease). Specimens required i. chronically exposed to arsenic (eg via arsenic contaminated feed over a long period or.25 Arsenic Urine mg/kg (wet wt) < 0. Sample of foetal pericardial or pleural fluid submitted chilled for virus isolation and serology. Sections of lung and other organs with lesions. It is often difficult to demonstrate the presence of organisms in chronic lesions. Diagnosis Based on clinical examination supported by history and laboratory examination to define the specific cause. NB. ARTHRITIS AND POLYARTHRITIS Diagnosis Based on clinical signs. bacteriological. Aseptically collected joint fluid. pathological and serological examination. spinal cord and peripheral nerves in buffered formalin for histopathology. 50 g of suspect source material for toxicology. iii. ARTHROGRYPOSIS AND HYDRANENCEPHALY Causes include infections of Akabane virus. Affected. ii. iii. NB There is no routine serological test available for Erysipelas. Conversely. Specimens required to allow for a differential diagnosis if arsenic is not the cause. iv. Specimens required i. ASPERGILLOSIS See: Fungal infections' ATAXIA Diagnosis of causes of 'ataxia' requires a careful clinical examination to determine whether the condition is due to a nervous. Maximum concentrations of arsenic in tissues occur about eight hours after ingestion. ii. iii. Brain and spinal cord in buffered formalin for histopathology. a blood sample from the neonatal animal if it has not sucked. iii. the dam with Palyam virus disease and as inherited Diagnosis Based on clinical signs and gross pathology. At least 3 ml of serum.lesions is adequate qualitative confirmation. submitted chilled for chlamydial serology. in the past stock regularly treated in arsenical dips) may have liver levels up to 8mg/kg and milk levels to 1. VETERINARY LABORATORY MANUAL 37 . Arthrogrypotic animals may have CNS lesions only in the spinal cord. Sections of skeletal and heart muscle in buffered formalin for histopathology.5 2-100 Interpretation depends on the interval between exposure and sampling. arsenic poisoning has been confirmed in the animal.5 Arsenic Milk mg/kg (wet wt) < 0. with the joint capsule intact. unopened joints. in buffered formalin for histopathology. kidney mg/kg (wet wt) < 0. submitted chilled. submitted chilled for serum enzymology and biochemistry. iv.5 Toxic > 8* > 1. chilled. clinically normal stock. Aino virus. ii. or.25 Possibly toxic 0. skeletal. 50 ml of milk or urine for arsenic analysis (only where chronic exposure is suspected). Interpretation of tissue and secreta arsenic concentrations Analyte Sample Units Normal Arsenic Liver. Serum sample from the dam. free of cells and haemolysis. to be examined only if v. submitted chilled for serology. Acute and convalescent serum samples from affected sheep or cattle. Specimens required i.5-8 0. and animals that survive for 2 to 3 days may have liver levels as low as 2 mg/kg. iv. as well defects.

Scabs from the lesion submitted chilled for bacteriology. Often associated with the presence of immature liver fluke. iii. Usually associated with vulvitis in ewes. collect nasal swabs from up to 10 clinically affected pigs (preferably weaners and growers). taken from a freshly dead animal. bacteriology. Specimens required i. At least three (3) impression smears from cut surface of lesion for staining with specific fluorescent antisera. protrusion of tongue and numerous large haematomata in the frontal sinuses. BENIGN FOOTROT See: Footrot BETA MANNOSIDOSIS OF GOATS See: Mannosidosis VETERINARY LABORATORY MANUAL 38 . At least three (3) smears of haemorrhagic muscle exudate for bacteriology. and submitted chilled for bacteriology.v. so that demonstration of the organism is not always diagnostic. 30 g of liver and kidney. Specimens required Bloat must be diagnosed on history and gross pathology. aluminium-shafted swabs. but also reported in Dorset Horn and other breeds. Fasciola hepatica. Portion of affected muscle. Can be diagnosed clinically when large numbers of pigs with snout deformities are present. BABESIOSIS See: Tick fever BALANO POSTHITIS IN RAMS Occurs more commonly in the Border Leicester breed. ii. submitted chilled in separate copper free containers for copper analysis. Despatch swabs in transport media to reach the laboratory within 24 hours of collection. ii. Specimens required i. BLACKLEG Diagnosis Must be based on a careful post mortem examination with demonstration of either Clostridium chauvoei or Clostridium septicum in the lesions of haemorrhagic myositis. so fresh selective media can be available in the laboratory. histopathology. • Use mini tipped. If bacteriological confirmation of toxigenic P multocida is needed. 'BIG KNEE' IN GOATS See: Caprine arthritis-encephalitis (CAE)' ATROPHIC RHINITIS OF SWINE Syn: Enzootic/progressive atrophic rhinitis Diagnosis Clinical signs. gross pathology. Diagnosis Demonstration and recovery of Cl novyi from necrotic foci. Specimens required i. iii. Snout swabs transported at ambient temperature in Amies Charcoal transport medium. Diagnosis Clinical findings. Portion of affected muscle in buffered formalin for histopathology. Specimens required These must be taken from a freshly dead animals: i. in buffered formalin for histopathology. Portion of liver. • Advise RVL of intended sampling. collected aseptically. ii. BLACK DISEASE Clostridium novyi is present soon after death in a large proportion of livers of normal sheep and cattle. post mortem findings. submitted chilled for bacteriology. including injection of eyes. Portion of liver containing lesions. especially when sampling weaner and grower pigs. BLOAT Diagnosis Clinical examination. If possible: • Swab nares with alcohol to reduce contamination. bacteriology in early cases to assist in attempting to determine aetiology. Swabs from the lesion submitted in Amies transport medium for bacteriology.

submitted chilled on the clot. Affected lambs submitted whole foetal membranes are also required if it is an anteparturient or parturient death. Suspect food material (approx 30g) for toxin ELISA. iv. which may or may not show nervous signs. Portions of fresh lung. AI centres select against BLAD heterozygotes when screening bulls for entry to their centres. Heparinised blood sample for antigen ELISA. which include flaccid paralysis. enterotoxaemia. liver. fixed in buffered formalin for histopathology. BLUE-GREEN ALGAL POISONING The toxic blue green algae. v. Serum from affected animals for serum chemistry/enzymology iii. The disease is characterised by progressive leucocytosis emanating from a deficiency of an adhesion factor on the surface of leucocytes that is essential for leucocytes to migrate from blood vessels to sites of infection.nsw. Specimens required i. bladder and nasal mucosa in buffered formalin for histopathology. BOTULISM There are five (5) major types of botulism (A to E). small intestinal contents and large intestinal contents for toxin ELISA. Specimens required i. Frozen serum. Types C and D are the usual causes of botulism in cattle. Sections of liver and kidney in buffered formalin. type C in horses and wild birds. BOVINE LEUKOSIS See Enzootic bovine leukosis' BOVINE LEUCOCYTE ADHESION DEFICIENCY (BLAD) (Holstein/Friesian calves) This defect was disseminated throughout the international Holstein herd from a founder bull in the USA whose descendants have been used extensively in the international Holstein herd.Specimens can be taken to eliminate other causes of sudden death.gov. iii. and the absence of gross and histopathological lesions.dpi. Consequently. Specimens required See http://www. Important cases may be confirmed by the BOVINE MALIGNANT CATARRH Syn: Malignant catarrhal fever Diagnosis Sporadic incidence. Blood sample. Specimens required i.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot. ii. 500 ml of scum and water for examination for toxic algae. Diagnosis Presence of algae as a green scum on the water. ii. Demonstration of Pestivirus in affected lambs and antibody in the ewe. liver and gut contents. 20 to 25 hairs. stomach contents. Evidence of a hepatotoxin and gastrointestinal irritation.pdf For diagnosis of the disease and for heterozygote detection: i. affected calves suffer with repeated bacterial infections. Brain and sections of sections of liver. spleen and mesenteric lymph node collected aseptically and kept chilled for antigen ELISA or virus isolation. kidney. The clinical syndrome includes sudden death or jaundice and photosensitisation. clinical signs. from the distal end of the tail. adrenal gland. Diagnosis Usually based on clinical signs. PCR for ovine herpesvirus2 (OHV-2). Specimens required i.g. BORDER DISEASE IN SHEEP Diagnosis Clinical signs including abortion and the birth of lambs with abnormally hairy birth coats. demonstration of the toxin in serum. hypomagnesaemia. histopathology. from the affected lamb and its mother for serology. e. Microcystis cyanea and Anabaena circinalis are mainly a problem in late summer and autumn. Brain and spinal cord. with roots attached. 39 VETERINARY LABORATORY MANUAL . ii.

submitted chilled for bacteriology and serology. 5 ml of EDTA blood for PCR testing for ovine herpesvirus-2 (OHV-2) iii. histopathology. submitted chilled for bacteriology and histopathology. submitted chilled for bacteriology. affecting the haematopoietic system and bladder wall. Specimens required (for cattle) Live animal i. VETERINARY LABORATORY MANUAL 40 . Uterine discharges or vaginal mucus. submitted in buffered formalin. BOVINE VIRUS DIARRHOEA See: Pestivirus BRACKEN FERN POISONING See also: Enzootic haematuria of cattle In ruminants. Serum samples submitted chilled for serological testing. Diagnosis History and serology. red femoral shaft bone marrow and affected tissues in buffered formalin for histopathology. Specimens required i. a progressive retinal degeneration. ii. Semen samples collected aseptically and submitted chilled for bacteriology. Serum sample. and submitted chilled for bacteriology. There is no specific diagnostic test in the live animal for bracken fern poisoning. Specimens should be submitted to eliminate other diseases. Abortion in ewes is not a common finding. In adult sheep. chilled for phosphorus estimation (see Hyophosphataemia). The diagnosis in rams must be based on the result of a clinical examination and serology. alternatively submit spleen and kidney chilled for pestivirus. Milk samples from the lactating cow. 2 ml of serum. including ampullae and seminal vesicles. Ocular and nasal swabs submitted in phosphate buffered gelatin saline (PBGS) for examination for infectious bovine rhinotracheitis (IBR). iii. ii. Specimens required BRUCELLOSIS BOVINE NSW reached Bovine Brucellosis free status in 1992. as there is a tendency for infected pigs to cease to react although they still remain infected and false positives can also occur. Sections of liver. principally a disease of cattle. Aborted foetuses iii. Testes. submitted chilled for serology. iv. Specimens required Cows i. Bulls i. for histopathology. iii. Blood films for haematology. 5 ml of blood in EDTA for haematology. The agglutination test is of use mainly as a herd test. kidney. epididymes and accessory sex glands. Diagnosis Clinical signs. 2 ml of serum for serology. ii. Samples can be submitted on the clot if they will be tested within 48 hours of collection. Samples can be submitted on the clot if they will be tested within 48 hours of collection. haematology. In dead animals. ii. From cases of haematuria. ii. Specimens as required for diagnosis of abortion in cattle (see Abortion in cattle). long term intake of bracken fern by adult sheep can induce "bright blindness". supported in some cases by bacteriological examination of semen.ii. BRUCELLOSIS PORCINE Brucella suis infection was last diagnosed in NSW in 1968. but associated with nervous signs from thiamine deficiency attributed to a thiaminase in the plant. Dead animal i.5 to 1 ml of semen collected aseptically. sections of bladder wall with lesions. pigs and other monogastrics rarely encountered. 5 ml of blood on clot submitted chilled for examination for pestivirus (mucosal disease). 0. iii. In horses. BRUCELLOSIS OVINE Diagnosis Characterised by infertility in rams due to epididymitis.

Abortion and infertility due to C fetus subsp venerealis can be confirmed by demonstration of specific antibodies in the vaginal mucus of affected cows by ELISA. Vaginal mucus samples collected for demonstration of C fetus subsp venerealis IgA antibodies by ELISA test. Abortion investigation: animals which have aborted can be sampled from 1 week to 3 months after the abortion. CAMPYLOBACTER ABORTION OF SHEEP Usually attributed to Campylobacter fetus subsp fetus (formerly subsp intestinalis). submitted chilled for bacteriology. C fetus subsp venerealis can also be isolated from the prepuce of infected bulls. lambs. Specimens required VETERINARY LABORATORY MANUAL 41 . chilled small intestine for bacteriology. CAMPYLOBACTER ENTERITIS Campylobacter jejuni and occasionally Campylobacter coli have been associated with enteritis and diarrhoea in calves. Calculi for chemical analysis. formerly bovine vibriosis Bovine venereal campylobacteriosis (BVC) is caused by Campylobacter fetus subsp venerealis and is characterised by infertility and early embryonic death. Serum samples (not blood) submitted chilled for serology (SAT and RBT). Diagnosis History of reduced breeding efficiency in herds where natural breeding is practised.i. Specimens required Cows i. ii. C fetus subsp venerealis and C fetus subsp fetus can be isolated from aborted foetuses.dpi. CAMPYLOBACTERIOSIS OF CATTLE Syn: Bovine venereal campylobacteriosis (BVC). Abortion occurs in a small percentage of infected cows. The role of other Campylobacters such as C sputorum and C hyointestinalis in the aetiology of diarrhoea and intestinal pathology in pigs is uncertain (see Porcine proliferative enteropathy). PBST diluent and instructions for collection of vaginal mucus samples for BVC ELISA test are available from the laboratory. dogs. Typical liver lesions in some full term lambs. Chemical analysis of the calculus may indicate the cause of the calculus formation. Formalin-fixed small intestine for histopathology. Demonstration of calculus at critical location. ii. poultry and man (C coli). occasionally Campylobacter jejuni implicated. Campylobacter fetus subsp fetus (formerly subsp intestinalis) also causes abortion in cattle.pdf ii. Clinical signs commonly observed are repeated returns to service. with prolonged interservice intervals and sporadic occurrence of mid to late term abortions. Diagnosis should be made on clinical and post mortem findings.nsw. Aborted foetuses submitted whole for bacteriology i. However. In a herd situation. CALCULI Diagnosis Evidence of uraemia/urinary retention. Specimens required i. iii.au/agricultur e/vetmanual/specimens-bydiscipline/bacteriology/01-bvc-elisaguide. poultry and man. demonstration of antibody in vaginal mucus is the preferred method of diagnosis as it can be difficult to recover the organism from the prepuce. histopathology. NSW Department of Primary Industries outsources this testing. Specimens required Specimens as required for the diagnosis of abortion in sheep. both may be found in otherwise normal intestinal tracts of sheep and cattle (C jejuni) or pigs. Diagnosis Late term abortions or birth of weak or dead full term lambs. Fresh.gov. Diagnosis Bacteriology. Bacteriology. ii. See http://www. cystitis and/or swelling and cavitation of kidney pelvis with pressure effects on cortex and medulla. pigs. Lesions.

See http://www. Affected calves can be identified at birth by their distinctive woolly haircoat. Infertility investigation: representative sampling of herd. Microscopic examination is necessary to accurately demonstrate the extent of the heart muscle changes. Any woolly-coated Poll Hereford calf that dies during the first few months of life should have a postmortem examination to check for evidence of cardiomyopathy. Portions of affected tissues in buffered formalin for histopathology (carpal joint. This can be done when pregnancy testing reveals infertility. Owners occasionally see an affected calf collapse and die. clinical history of nervous signs in young kids.gov. ill thrift.nsw. ‘wirey’ or ‘curly’). 1 x 10 ml lithium heparin blood for glutathione peroxidase (GSHPx). Diagnosis Clinical signs. CWH is a congenital genetic disease.5 mg EDTA/ml of blood) for DNA isolation (from affected calf. dam and sire). hard udder and mastitis. Bulls i. CAPRINE ARTHRITIS ENCEPHALITIS (CAE) Syn: ‘Big infection knees’. usually following exertion Some CWH calves show clinical signs of progressive heart failure. CARDIOMYOPATHY AND WOOLLY HAIRCOAT (CWH) SYNDROME (Poll Hereford calves) Dd: Cardiac white muscle disease (nutritional myopathy. liver. with an autosomal recessive mode of inheritance presumed to involve defective structural protein(s) within desmosomal complexes. histopathology of affected tissues. tongue. and gross cardiac ventricular fibrosis. Preputial scrapings submitted in Campylobacter transport enrichment medium (TEM) for bacteriology. CAMPYLOBACTERIOSIS OF PIGS See: Porcine proliferative Campylobacter enteritis' enteropathy. Fresh liver and kidney for selenium estimation. Specimens required i. Some CWH calves have protruding eyes and keratoconjunctivitis (‘pinkeye’). ii. history. Serum samples chilled for viral serology. iii. 42 . 2 x 10 ml of EDTA blood (1. These DNA samples will be held pending development of a diagnostic DNA test (not yet available). Most calves with CWH die suddenly. brain. lung). spinal cord. iv.pdf Aborted foetuses i. Death from heart failure occurs between birth and 12 weeks of age. It is not sufficient to diagnose CWH in a woollycoated calf without postmortem confirmation of the cardiomyopathy. Whole heart. Specimens required i. Specimens as required for diagnosis of abortion in cattle (see Abortion in cattle). ii. chronic progressive pneumonia. Calves with CWH are not selenium-deficient. with or without mineralisation. psoas muscle) in buffered formalin. particularly if the woolly haircoat is overlooked. big knees in adults associated with lameness. Specimens required i. nutritional myopathy). weight loss.dpi. selenium deficiency). CWH cases can be misdiagnosed as selenium deficiency (cardiac white muscle disease. (also described as ‘fuzzy’. Cardiac arrhythmias are usually detectible. VETERINARY LABORATORY MANUAL CASEOUS LYMPHADENITIS (CLA) Diagnosis Recovery of Corynebacterium pseudotuberculosis from lesions. at least 10 vaginal mucus samples collected from infertile heifers or cows. kidney and skeletal muscle (including diaphragm.au/agricultur e/vetmanual/specimens-bydiscipline/bacteriology/02-bull-vdcoll. Caprine retrovirus Diagnosis Viral serology.iii. Smears from early lesions or from the periphery of active lesions for bacteriology. eyes. TEM is supplied from the laboratory together with instructions for collection of samples.

Diagnosis Based on clinical findings with demonstration of group specific antibody rise between 'acute' and 'convalescent' serum samples in association with disease. The mutation is now relatively rare in the Australian Holstein herd due to selection against heterozygotes by AI centres. Heterozygote detection i. histopathology. Portions of tissues containing early lesions or from the periphery of active lesions. e. ii. Response to cobalt supplements or injections of vitamin B12. from each of 3 to 5 animals in the affected group. 10 ml of blood. Specimens required i. specimens as required for the investigation of nervous disorders.ii. reproductive performance of older ewes in the flock and demonstration of cystic endometritis. Diagnosis Clinical signs. ii. deficiency of other minerals.gov. clotted in vacuum tubes. is considered significant.dpi. chewing on fixed objects. submitted chilled for bacteriology. Specimens required i. sporadic bovine encephalomyelitis (SBE). within 3 to 4 weeks of the clinical episode. submitted chilled for biochemistry. chlamydial arthritis. CHLAMYDIAL INFECTIONS Syn: Chlamydiosis. chlamydial abortion of sheep. Serum samples taken in the acute and convalescent stages of disease (3 to 4 weeks apart) from individually identified and observed animals. Portions of tissue containing early lesions or from the periphery of active lesions. The mutation responsible for this defect was imported into Australia by a US-born bull whose descendants were selected for high EBVs for butter fat content of milk. submitted chilled for chlamydial serology. confirmed by DNA analysis. Interpretation of Titres The demonstration of a four fold rise in CFT antibody titres. arthritis or abortion. VETERINARY LABORATORY MANUAL 43 . with roots attached. Diagnosis History of ill thrift. weaned animals.pdf (In order of preference) Diagnosis of the disease i. Reproductive tracts including ovaries from 15 to 20 ewes which did not lamb at the previous lambing. Demonstration of Chlamydia by FAT on smears of fresh tissues or fluids. head pressing and ultimately collapse and death between the third and fifth day of life. Submit chilled. CLOVER DISEASE Diagnosis Based on flock history. as it is often difficult to determine endometrial cysts grossly in fixed tissue. causing serious illness and sometimes death in humans. from the distal end of the tail. Depending on the clinical condition.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot. parasitism. in young animals particularly after elimination of other likely causes. with roots attached. cobalt deficiency is confined to restricted geographical areas. Evidence of a mild to moderate macrocytic anaemia associated with weepy eyes. Signs have been seen in sheep and cattle. 20 to 25 hairs. become depressed by 24 hours. 20 to 25 hairs. Whole brain in buffered formalin for histopathology. preferably in young. iii. Specimens required See http://www. Typical clinical signs are seemingly aimless wandering. in buffered formalin for histopathology. COBALT DEFICIENCY In NSW. tongue protrusion. iii. malnutrition.g. Specimens should be submitted to eliminate other causes of ill thrift. CITRULLINAEMIA (Holstein/Friesian calves) Affected calves are clinically normal at birth. avian chlamydiosis Caution Chlamydiosis is a zoonotic disease. develop signs of neurological dysfunction by 48 hours. from the distal end of the tail. Specimens required ii.nsw.

goats and pigs.K88ac: 0141:K85ac: 0141:K85ab: H10 H4 or HNM H4 or HNM Neonatal mortality + Neonatal diarrhoea + Weaner mortality + + + Weaner diarrhoea + + + Weaner nervous signs + + Weaner oedema disease + + 08:K87. cattle. Specimens required i. oocysts may be difficult to detect in faeces during the diarrhoeal phase. jejunum) showing lesions in buffered formalin for histopathology. ii. liver. Portions of intestines (e. Demonstration of pathogenic strains of E coli. ii. anaemia and ill thrift. Oocysts of E stiedae may be demonstrable in the exudate of dilated bile ducts and gall bladder COLIBACILLOSIS See also: Oedema disease. Sections of liver. histopathology. Specimens required i. Faecal sample for oocyst count. iii. 44 COMPLEX VETERBRAL MALFORMATION (CVM) Affected calves are generally aborted. Diagnosis Clinical signs. In pigs. Portion of affected liver in buffered formalin for histopathology. small intestine and mesenteric lymph node submitted in buffered formalin for histopathology. brain submitted in buffered formalin for histopathology. liver with multiple yellow or white nodular lesions. The VETERINARY LABORATORY MANUAL . In pigs.COCCIDIOSIS Occurs mainly in young sheep. Diagnosis Gross pathology.K88ac: H19 or HNM + + + + POULTRY There is no real information on the prevalence of pathogenic serogroups in poultry in Australia. kidney. but in rare instances may be born alive. colon and mesenteric lymph node. iv. In acute coccidiosis. demonstration of large numbers of coccidia in faeces (chronic). Typing of poultry clinical isolates is not being undertaken at this time.g. iii. Characterised in ruminants by scouring. enterotoxaemia or septicaemia. and a definitive diagnosis can be made by examination of small intestinal smears of a typically affected pig. ii. obtain 5-10 impression smears of ileum and jejunum and air dry. impression smears of intestine (acute). COCCIDIOSIS – HEPATIC IN RABBITS Characterised by mortalities with gross pathology typically revealing an enlarged E coli serotypes commonly associated with colibacillosis in pigs in NSW 0149:K91. Demonstration of Eimeria stiedae. Preferably the whole animal submitted chilled. and other animals with acute coccidiosis. The mutation responsible for this defect has been traced to a founder US Holstein sire whose descendants have been used extensively worldwide. In piglets. AI centres select against CVM heterozygotes when screening bulls for entry to their centres. scouring Diagnosis Bacterial evidence of Escherichia coli enteritis. In animals showing nervous signs. for bacteriology. Specimens required i. and histopathology. it has been shown to be an important cause of scouring in 10 21 day old suckers in some herds. Sections of lung. disease is characterised by shortening of the cervical and thoracic vertebra and symmetrical arthrogryposis in the fore and sometimes rear legs. Lesions occur in the mucosa of the intestinal tract. kidney. submitted chilled. ileum. oocysts are often not yet present in faeces. jejunum. and impression smears of affected intestine and histopathology are recommended.

demonstration of virus particles on electron microscopy. Pregnant mares .nsw. submitted in a sealed container.dpi. It is important to eliminate infectious causes before incriminating environmental. i. abortion. Specimens required i.3 swabs per animal. orf Diagnosis Clinical signs. COPPER DEFICIENCY Diagnosis History. a Macartney bottle. the occasional abnormality or freak is of little interest and there is no benefit in submitting such cases. from cases of suspected enzootic ataxia. Specimens required i. submitted chilled for serology and examination for immunoglobulins. e. In general. submitted chilled for serology. The breeding history of the animals may be important in some cases. clotted in vacuum tubes. for electron microscopy. CONGENITAL ABNORMALITIES See also Akabane. NB In all cases of suspected copper deficiency. serum and liver copper levels. 10 ml of blood. iii. However. from each of 3 to 5 animals in the affected group. Stallions 3 swabs per animal. Non-pregnant mares . together with foetal membranes if available.gov. ii. Specimens required Sample collection equipment must be free of copper. Brain and spinal cord. Specimens required Swabs should be taken from each of the following sites and submitted without VETERINARY LABORATORY MANUAL . 45 CONTAGIOUS EQUINE METRITIS (CEM) Diagnosis Recovery of Taylorella (formerly Haemophilus) equigenitalis from the genital tract. arthrogryposis and hydranencephaly. from the following sites: • cervix • clitoral sinus • clitoral fossa iii. Serum sample from mother of affected animal. Body fluid (preferably pericardial or pleural) from the foetus or serum sample from the neonate prior to sucking. ii.g. Vacuum blood collection tubes are satisfactory for blood testing.Specimens required See http://www.pdf Diagnosis of the disease and heterozygote detection i. increased incidence of congenital abnormalities or repeated occurrence over a number of years warrant further investigation. Scabs with underlying tissue scrapings.2 swabs per animal from the following sites: • clitoral sinus • clitoral fossa CONTAGIOUS OPHTHALMIA See: Ophthalmia CONTAGIOUS PUSTULAR DERMATITIS Syn: Scabby mouth. genetic diseases. chemical or other teratogenic causes or inherited defects. clinical signs. contagious ecthyma. response to therapy. 20 to 25 hairs with roots attached. particularly in relation to events during the first half of pregnancy may be helpful. from the following: • prepuce and base of penis (smegma) • urethral fossa and urethral sinus • terminal urethra ii. Differentiate from Staphylococcal infections and Dermatophilosis. submitted chilled for biochemistry. Affected animals submitted whole. From the distal end of the tail refrigeration in Amies charcoal transport medium for bacteriology: i. clotted blood is preferred to liver or kidney.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot. Diagnosis Clinical signs. A detailed history. pestivirus. in buffered formalin. Advice should be sought from the laboratory in these cases.

The terminal acute phase is characterised by liver necrosis. Disease is spread by infectious oocysts. Identification of organisms in the brush border of mucosal surfaces on histopathology. Interpretation of copper concentrations (cattle and sheep) Analyte Sample Minimum Special Units Species amount requirement Copper Liver 30g Fresh mg/kg Cattle (wet wt) Sheep Copper Kidney 30g Fresh mg/kg Cattle. (wet wt) sheep Toxicological results must be interpreted in relation to history. birds. haemoglobinuria and jaundice. Calf milk replacers should contain <20 mg copper / kg. Diagnosis Clinical signs. are coccidial parasites that are not host specific.5-20 COPPER POISONING OF SHEEP AND CATTLE See also: Pyrrolizidine alkaloidosis' Chronic copper poisoning in ruminants of the hepatogenous and/or phytogenous form is quite common. guinea pigs and other mammals. and man are common hosts. See Toxicology for suggested normal and toxic values.Interpretation of copper concentrations (cattle and sheep) Analyte Sample Minimum Special Units amount requirement Copper Serum or 1 ml OK on clot µmol/L hep plasma Species Cattle Sheep Deficient < 7. piglets. The diagnosis can be confirmed histologically. are not susceptible to conventional anti-coccidial treatment and to date have no available efficacious specific chemotherapy.5-16 7. easier and cheaper to perform than tissue analysis. Host debilitation and/or concurrent viral or bacterial infection may predispose to infection. although CORYNEBACTERIUM EQUI IN HORSES OR PIGS See: Rhodococcus equi pneumonia in horses See: Rhodococcus equi lymphadenitis in pigs' CRYPTOSPORIDIOSIS Cryptosporidium spp. histopathology. typical post mortem findings. dogs. Antigen capture ELISA on faeces of calves (where available). haemolytic anaemia. rabbits. clinical findings and histopathology.5 < 7. Deficient <4 <4 Normal 20-70 40-70 4-6 Elevated > 100 > 200 >8 associated with gastrointestinal or respiratory disease. kids. Signs most commonly seen are anorexia and diarrhoea in calves. Ruminants. by examination of faecal smears with acid-fast stains or phase contrast microscopy of faecal flotations. Liver concentrations of 200 mg/kg (wet wt) can be found in sheep dying from other causes. Sheep of all ages and preruminant calves accumulate copper in the liver when dietary copper is in excess of 30 mg copper /kg dry weight of feed consumed (adult cattle require and therefore tolerate 100 mg copper /kg of feed). foals and deer calves. in buffered formalin for histopathology. Specimens required i. Sections of liver & kidney. and have high environmental resistance. horses. ii. submitted chilled and unpreserved for copper estimation. especially sheep affected with pyrrolizidine alkaloidosis. as well as in reptiles and fish. They are VETERINARY LABORATORY MANUAL 46 . are quicker. Histochemical methods for staining for copper in liver sections although not quantitative. affecting particularly young animals. lambs. Diagnosis Identification of oocysts in faeces. Immuno-compromised or immuno-incompetent animals are at high risk of infection. mice. Occasionally detected in young cats. in separate copper free jars. NB.5 Normal 7. which are shed in faeces or respiratory secretions. 30 g kidney and liver. analysis of tissues. which promotes liver copper accumulation. Kidney is considered superior to liver for biochemistry in acute and chronic copper poisoning: Kidney copper concentrations rise faster in acute toxicosis. pigs.

or histopathology of degenerate cysts. It is not known if exists in the Australian Holstein/Friesian herd. iii. Specimens required See http://www. ii.this technique may be less sensitive than microscopic examination. Faecal sample for preparation of smears and examination by MZN or similar stain. formalin-fixed small intestine. The mutation responsible for the defect has been recorded in North American Holsteins. iii. Specimens required i. The causal organism may be isolated from scrapings or a biopsy specimen. Specimens required i.dpi. Specimens required Plant specimens can often be identified by the local District Agronomist. post mortem appearance of blood and mucosae. for histopathology DEFICIENCY OF URIDINE MONOPHOSPHATE SYNTHETASE (DUMPS) (Holstein/Friesian cattle) Deficiency of uridine monophosphate synthetase results in foetal death during the first trimester of pregnancy. Fresh and formalin-fixed specimens to establish an alternative diagnosis when necessary DERMATOPHILOSIS Syn: Mycotic dermatitis. Cyanogenetic glycosides in plants hydrolyse during transit so that laboratory analysis is often misleadingly negative. with roots attached. CYSTICERCOSIS Diagnosis DIARRHOEA See Scouring VETERINARY LABORATORY MANUAL 47 . in a sire line that.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot. or by floatation and examination by phase contrast microscopy. Specimens required i. Blood smears to check for nitrate poisoning. See Poisoning (plants) for information on collection of specimens for plant identification. Samples of suspected plant material for identification and check for cyanide and nitrate. reptiles or fish. ‘lumpy wool’ Diagnosis Gross appearance of the lesions and demonstration of Dermatophilus congolensis microscopically. iv. caecum. submitted frozen for toxicology. was not used widely in the international Holstein herd.nsw. or epithelium of the upper respiratory tract and lung from freshly-killed cases. fortuitously. i. CYANIDE POISONING Syn: Prussic acid poisoning. iii.gov. Immersion in 1. chilling is not sufficient. Formalin-fixed abomasum/stomach (peptic gland epithelium) and small intestine from freshly-killed mammals.3% mercuric chloride also stops hydrolysis. Skin biopsy from the area with lesions.pdf Heterozygote detection i. Sample of wool or hair showing lesions or scabs for gross examination and demonstration of D congolensis in impression smears. ii. submitted chilled for bacteriology. Skin biopsy from the area with lesions. in buffered formalin. Gross pathology and dissection of a viable scolex from the fluid filled cyst. 20 to 25 hairs. Submit frozen for cyanide toxicology. Diagnosis Clinical signs. Sample of rumen ingesta in an airtight container. odour of bitter almonds. chemical demonstration of cyanide in plant material or rumen ingesta. from the distal end of the tail. Herbage and ingesta should be kept frozen from the time of collection to the time of analysis. ii. colon and bursa. Lesions. Fresh tissue with lesions. For birds. submitted chilled for parasitology. ii. in buffered formalin for histopathology. access to suspected materials or fodders.

DWARFISM IN DEXTER CATTLE Syn: Dexter chondrodysplasia. virus isolation. Please provide pedigree of the subject with the hair sample. with extreme shortening of limbs and vertebral column. kidney and lung. cleft palate and protruding tongue). vi. Sections of liver. Specimens as required for the differential diagnosis of nervous disorders. kidney. i. submitted chilled for virology. Sections of heart. gross craniofacial defects (relatively large head with retruded muzzle. in buffered formalin for histopathology. iv. typical white areas on heart and throughout myocardium. kidney. submitted chilled for bacteriology. Specimens required i. Diagnosis Clinical signs. Brain and/or spinal cord if any neurological signs evident. and a large abdominal hernia. ii. Specimens required See http://www. clinical signs and in some cases. Congenital lethal chondrodysplasia (Dexter bulldog) is the homozygous form of Dexter chondrodysplasia. Serum samples for virus serology. Specimens required ii. cattle.DRENCHING MORTALITIES Diagnosis Based on history. iii.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot. The disease is a zoonosis affecting a wide range of mammals including mice. • VETERINARY LABORATORY MANUAL 48 .pdf Heterozygote detection i. spleen and whole brain in buffered formalin for histopathology. ENCEPHALOMYOCARDITIS (EMC) OF PIGS Encephalomyocarditis is caused by a picornavirus virus. Specimens required Product information sheets from the manufacturer should be checked to ensure the appropriate specimens are submitted. Sample of cerebrospinal fluid collected aseptically. histopathology. lung and heart blood submitted chilled for bacteriology. in buffered formalin for histopathology.dpi.nsw. porcine enterovirus encephalomyelitis. 20 to 25 hairs. ii. lung. v. liver. please submit specimens as required by the National TSE Surveillance Program (http://www. Section of liver. Fresh heart and spleen collected separately and aseptically submitted chilled for virology.au/pr ograms/adsp/adsp_home. Diagnosis History of sudden death. transmissible spongiform encephalopathy (TSE) For adult sheep and cattle with progressive neurological disease.cfm).com. Virology and bacteriology to establish an aetiological diagnosis.animalhealthaustralia. with roots attached. Section of brain and spinal cord collected aseptically. congenital lethal chondrodysplasia (Dexter bulldog). Associated with abortion in some instances. accompanied by a completed NTSESP ENTEROTOXAEMIA Difficulty is often encountered in establishing a diagnosis of enterotoxaemia in sheep. wombats and elephants.gov. iii. Brain and portion of spinal cord. The following points must be considered: • Clostridium perfringens is a normal bowel inhabitant and toxin may be present in the gut content of healthy animals. Affected calves are usually aborted before the seventh month of gestation. spleen. Other samples as required from the product information sheet of the manufacturer Clinical History and Post-mortem Report form. goats and deer. histopathology. Cl perfringens is a common postmortem invader of parenchymal tissues and will be present by 8 hours after death. ENCEPHALOMYELITIS AND ENCEPHALITIS See also Haemagglutinating encephalitis virus (HEV) of pigs. from the distal end of the tail.

Diagnosis A positive diagnosis rests on several of the following being present: • History of sudden death • Necropsy findings: • glycosuria • pulmonary oedema and congestion • fibrin clot in pericardial sac • epicardial haemorrhages • yellow creamy intestinal contents NB: some animals may show minimal gross changes e. Typical kidney lesions develop in sheep within a few hours of death. The virus has been eliminated from dairy herds in NSW but beef cattle remain infected at a low prevalence. for glycosuria should be carried out in the field. Like other retrovirus infections (AIDS. Gross pathology histopathology of the urinary bladder. Congenital transmission also occurs but is less important.. Sections of liver and kidney. and and Specimens required i. In chronic cases. toxin is usually no longer present. Formalin fixed sections of affected lymph nodes.• • • Toxin may diffuse from the gut if the autopsy is delayed. etc. EIA) antibody indicates current (not just previous) infection.g. submitted in buffered formalin for histopathology. Diagnosis Clinical signs. particularly from areas where the contents are a yellow colour and creamy consistency. they are not present at the time of death. Specimens required i. confirmed by virus serology. e. Serum or preserved milk samples (bromopol preservation as used for milk cell counts) for virus serology (ELISA or GDPT) ii. in focal symmetrical encephalomalacia. At least 20 smears from the intestinal mucosa at sites from the abomasum to the colon with particular attention to the ileum. Urine examination VETERINARY LABORATORY MANUAL ENZOOTIC HAEMATURIA OF CATTLE See also Bracken fern poisoning Diagnosis Chronic intermittent haematuria anaemia. NB Indicate vaccination history with all submissions of suspected enterotoxaemia. and whole brain in buffered formalin for histopathology. thymus. sudden death with some small intestinal inflammation. Submit chilled for toxicology. ii. ENZOOTIC ATAXIA Syn Copper deficiency' ENZOOTIC BOVINE LEUKOSIS (EBL) A retrovirus infection of cattle which can be spread by blood transmission from rectal palpation. CAE. • histopathological lesions in brain and kidney (e. spleen. demonstration of protein leakage from blood vessels into perivascular spaces of cerebrum). ENZOOTIC PNEUMONIA OF PIGS See Porcine enzootic pneumonia' 49 .g. tattooing or gouge dehorning where repeatedly-used equipment is employed. with particular attention to the ileum. Type D toxin is stable in gut contents in vitro without preservative. but other toxins are very unstable and have disappeared within 12 hours regardless of preservation. Sections of urinary bladder with suspect lesions. iii. cooling.g. • abnormal numbers of Cl perfringens like organisms in smears of intestinal mucosa taken from multiple sites from abomasum to colon. 4-5 sites can be smeared on one slide. Affected animals usually develop antibody within 2-3 months of spread. Specimens required i. histopathology and haematology. intestine or other organs showing gross lesions for histopathology. Samples of small intestinal content at least 10 ml from sheep and 40 ml from cattle should be collected from several sites in the small intestine. vaccination. • demonstration of epsilon toxin in gut contents by counterimmunoelectrophoresis (CIEP). Sites selected should be from areas showing haemorrhages and also from nearby non inflamed areas which may show a heavier flora.

NB. Specimens required i. Haematology. Specimens required i. submitted chilled for bacteriology and histopathology. iii. Diagnosis Clinical signs of depression and anorexia with intermittent fever. At least 2 ml of serum for viral serology (AGID . haemoglobinuria and subcutaneous oedema may be present. Diagnosis Demonstration of Mycoplasma ovis and anaemia in sheep. Actinobacillus seminis. EPIDIDYMITIS See also Brucellosis-ovine. Paired serum samples (with a 2 to 3 week interval) from individually identified animals collected in the very early acute and convalescent phases submitted chilled or frozen for serology. ii. Specimens required i. kidney. Blood films for haematology. The presence of the organism is insufficient to establish the diagnosis as it can be found in normal sheep. however. the AGID may occasionally be negative early in the disease course.Coggins test). Blood films from 10 affected and at least 20 apparently normal animals in the flock for examination for M ovis (See Specimens for Haematology . serology. Serum sample.preparation of blood films'). Specimens required i. Thick blood smears dried under cover (not by heat or exposure to sunlight) for parasite examination. Generally virus serology is not carried out unless paired serum samples are submitted. Histophilus ovis Diagnosis Clinical signs and presence of lesions. Recovery of causal organism from semen or reproductive organs. iii. Genital organs. Other causes of anaemia must be excluded. Semen sample collected aseptically. At least 2 ml of serum for serology (B equi IFAT). Brucella ovis. Sections of liver.EPERYTHROZOONOSIS OF SHEEP The taxonomy of the causative organism has been revised and it is now known as Mycoplasma ovis. The disease probably no longer exists in Australia. EQUINE INFECTIOUS ANAEMIA (EIA) Diagnosis Clinical signs of recurrent febrile attacks associated with anaemia over a period. Demonstration of the organism in thick and thin blood smears. Virus antibody rise between acute and convalescent serum samples. ii. 5 ml of blood in EDTA vacuum tube and blood smears for haematology. Clinical disease has not yet been recorded. submitted chilled for bacteriology. ii. iv.g. Specimens required i. 5 ml of blood in EDTA vacuum tube for haematology. Blood samples from affected animals in EDTA vacuum tubes for haematology.Coggins test) and histopathology. ii. submitted chilled for serology. 50 . serology (AGID . EPHEMERAL FEVER Diagnosis General symptomatology and occurrence in the herd and district. VETERINARY LABORATORY MANUAL EQUINE VIRAL ARTERITIS (EVA) See also Exotic diseases' Recent identification of seropositive horses and a semen-culture positive stallion in Australia suggest this organism has been established in the horse population at a low level. Anaemia. iii. In chronic cases there may be loss of condition and mild anaemia. A positive AGID is conclusive evidence of EIA. Evidence of infection with specific pathogens. including the accessory sex glands. e. EQUINE BABESIOSIS This disease caused by the protozoon Babesia equi was introduced to Australia in infected imported horses and transmitted locally by poor injection technique. spleen and lymph nodes in buffered formalin for histopathology. Actinobacillus seminis and Histophilus ovis.

ii. fresh lung. tissues of aborted foetuses (lung. Serum ii. submitted chilled for bacteriology. The Officer manning this phone will advise on what actions need to be taken. Gross pathology. Whole joint unopened and chilled.EVA is usually subclinical. Portions of above tissues in buffered formalin for histopathology. The manuals are revised regularly and the current versions can be downloaded from the Animal Health Australia website at http://www. subcutaneous oedema of limbs and ventral abdomen. semen for virus isolation. kidney. virus isolation. Abortion As this is associated with maternal pyrexia rather than foetal or placental infection. On suspicion of an exotic disease. Septicaemia i. ERYSIPELAS Erysipelothrix rhusiopathiae causes arthritis (often post-marking) and post-dipping lameness (due to laminitis) in sheep and arthritis. collected aseptically and forwarded in separate sterile containers for bacteriology. Ausvetplan is a comprehensive set of manuals dealing with all aspects of exotic disease management in Australia. EXOTIC DISEASES Guidelines on the collection and submission of specimens for exotic diseases can be found in 'Exotic Diseases of Animals: A field Guide for Australian Veterinarians'.com. Virus is spread by respiratory and venereal transmission. 51 VETERINARY LABORATORY MANUAL . Isolation of E rhusiopathiae from affected tissues. i. Virus is present in semen. depression. rhinitis and conjunctivitis with nasal and ocular discharge and photophobia. Forman AJ and Nunn MJ (1999). bacteriological examination of aborted foetuses is usually of little assistance in the diagnosis of Erysipelas abortion. Diagnosis Serology. spleen. Blood containing anticoagulant. Urticaria and endocarditis EXUDATIVE EPIDERMITIS IN PIGS Syn: Greasy pig disease Diagnosis Typical skin lesions on face and body. lymph node). iii. From acute clinical cases. if so. Diagnosis Clinical signs and history. sample of citrated blood collected aseptically. From adult horses at necropsy. a skin rash. urticaria. ii. forward frozen on dry ice. spleen and lymph nodes draining oedematous areas. thymus. Sections of liver.au/. but clinical disease overseas is characterised by pyrexia lasting 1-5 d. Tissues from foetuses or adult horses as above. the exotic disease hotline phone number is 1800 675 888. fixed in neutralbuffered formalin for histopathology. Send chilled unless delays of more than 24 h are anticipated in transit. veterinarians must immediately notify NSW Department of Primary Industries. Up to 70% of pregnant mares abort during the febrile stage. Portion of lesion(s) in buffered formalin for histopathology. In some circumstances. The resultant pyrexia in sows can result in abortion. faeces and aborted foetuses. If this is not possible. histopathology. anorexia. respiratory secretions and also in urine. forward aseptically-collected synovial membrane (most desirable) and synovial fluid (of lesser value) in separate sterile containers. veterinarians on a suspect property will be asked to remain until appropriate arrangements can be made to collect samples and disinfect equipment and samples off the property. nasal swabs or washings in PBGS. spleen. Portion of lesion(s) collected aseptically and forwarded in sterile containers for bacteriology. Specimens required Arthritis i. liver. Isolation of Staphylococcus hyicus. Geering WA.animalhealthaustralia. lung. dyspnoea and diarrhoea. There is no routine serological test at present available for Erysipelas. valvular endocarditis and fatal septicaemia in pigs. Specimens required i. This comprehensive book was published by CSIRO Publishing and distributed to all Australian veterinarians. Affected animals usually recover. iii.

Demonstration of numbers of spores of Pithomyces chartarum on pasture.dpi. It would be reasonable to expect dehorning and parturition may also present lifethreatening challenges to homozygous heifers. some daughters were observed to bleed excessively when ear marked. submitted chilled for fungal examination. or into milk at the onset of lactation. Sections of liver and kidney in buffered formalin for histopathology. at parturition.gov.Specimens required i.pdf i. ii. The wool may be discoloured due to the growth of chromogenic bacteria. Diagnosis Clinical signs. Staphylococci and Streptococci are often present in the lesion. Dehorning would also present a life-threatening challenge to bulls that inherited the mutation from their clinically normal heterozygous dam. Specimens required i. Material from the depths of the lesion submitted chilled for bacteriology. Specimens required i. high humus containing pasture. In one herd when a “bleeder” bull was used over heterozygous cows. but it has often been associated with Brucella abortus infection. Skin biopsy from affected area submitted in buffered formalin for histopathology.5 cm of pasture growth and including the dead leaves and leaf residue. bacteriological and macroscopic examination of affected wool. with roots attached. abundant. FACTOR XI DEFICIENCY (Holstein cattle) An autosomal recessive defect reported in Holsteins in North America and Europe. humid weather conditions. The mutation has been VETERINARY LABORATORY MANUAL 52 . caused either by prolonged wetting of the skin or other skin damage. detected in Holstein bulls in Australian artificial breeding centres. FACTOR VIII DEFICIENCY (Hereford cattle) Syn: Haemophilia A. serology. The serous exudate results in matting of the wool. Specimens required See http://www. At least 1 kg of pasture taken from the lower 5 to 7. Serum sample from affected animal for Brucella abortus serology (RBT and CFT). from the distal end of the tail. The condition has been recorded in Hereford cattle in Australia. 20 to 25 hairs.nsw. FASCIOLIASIS IN SHEEP AND CATTLE See Liver fluke infection FISTULOUS WITHERS The aetiology is not fully known. Diagnosis History of extended bleeding following dehorning. The mutation responsible for the defect has been defined. Specimens required i. A sex-linked inherited defect that usually presents as a fatal hemorrhage event following castration. submitted for gross examination ii. Live or fresh chilled dead piglet for bacteriology and histopathology. Smears and swabs from skin lesions. Sample of affected wool. clinical signs. Diagnosis A history of severe haemorrhages in grandsons of a sire should raise a suspicion of haemophilia A. ii. submitted in Amies charcoal transport medium. but may arise in any breed due to new mutations. Diagnosis Clinical signs. gross pathology and histopathology. ii. FACIAL ECZEMA Diagnosis History of moist. FLEECE ROT A dermatitis of the skin.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot.

The lesions will persist under dry environmental conditions and can affect a high percentage of the mob. Where underrunning is evident.nsw.au/reader/sheepfootrot A. Smears are taken from an active site in the lesion this is at the junction of healthy and diseased tissue.nsw. Specimens as required to investigate nervous disorders in sheep. but the granulation associated with the initial infection will be found near the toe. More common in sheep that have been run in wet conditions for some time. Early lesions resemble benign footrot.au/reader/sheepfootrot/a0956. grazing cereal crops sown in ploughed ground. Diagnosis History and clinical findings.Footrot in sheep and goats.FOCAL SYMMETRICAL ENCEPHALOMALACIA (FSE) A chronic form of enterotoxaemia. Usually affects a hind foot. wet pastures). the horn of the hoof may have to be cut away to reveal the active sites. often tracking up to break out at the coronet in front of the foot.gov. FOOTROT IN SHEEP Includes virulent and benign footrot For the purposes of the Stock Diseases Act and the NSW Footrot Strategic Plan. and commonly affect the interphalangeal joint. Not all chronicallyinfected sheep show lameness. Outbreaks often seen 7-10 days after susceptible sheep have been run in wet. Usually more than one foot affected. Suppurative lesions break out in the heel area with discharging sinuses and granulation tissue. a 53 VETERINARY LABORATORY MANUAL . Specimens required i. AgFacts): http://www.gov. all forms of non-benign footrot are treated as virulent footrot. usually involving the front feet and often associated with shelly toe. In addition. but if severely challenged can affect 2 or 3 feet. When the diseased tissue is pared away. Bacteriology (clinical diagnosis may be reinforced by demonstration of D nodosus-like organisms in smears). Diagnosis History. VIRULENT FOOTROT IN SHEEP Caused by strains of Dichelobacter nodosus (formerly Bacteroides nodosus) with a range of virulence. muddy areas. Specimens required None. heavy sheep especially ewes in late pregnancy and rams. ii. Further information (Policy. For full discussion of footrot classification (including colour photographs) refer to AgFact . Specimens required FOOT ABSCESS IN SHEEP Including: Toe abscess. Occurs in fat. heel abscess (infectious bulbar necrosis) TOE ABSCESS Caused by a range of pyogenic bacteria. Little or no pus present even in severe lesions. false negative smears can occur.agric. but are unable to distinguish between virulent and benign strains. but progress to severe interdigital dermatitis and underrunning of the sole and hard horn of axial and abaxial wall within 3-4 weeks. (http://www.agric. The lesion involves soft tissues of the toe region. muddy conditions (wet muddy yards. Can occur in younger sheep if they are put in wet. Diagnosis Clinical signs. Brain in buffered formalin for histopathology. histopathology of the brain. clinical examination of a sufficient number of sheep in the flock (to detect underrunning of the hard horn of the hoof and to be confident of the range of lesions present). Lesions seen as swelling and tenderness in the affected claw with severe lameness. Outbreaks of severe lameness can occur under wet pasture conditions when the mean daily temperature exceeds 9-10oC. Affected foot may be deformed. best diagnosed in the field. This may take a little while to download. HEEL ABSCESS Caused by infection with Fusobacterium necrophorum in association with other bacteria. Toe abscess can also cause underrunning and break out near the heel. Smears of affected feet lesions for D nodosus detection Smears of affected feet can be used to detect the presence of D nodosus. i.pdf?).

a distinction between virulent and benign can be made provided there are no recent treatments or environmental conditions which would mask the full expression of disease due to the infecting strain of D nodosus. but does not progress to underrunning of the hard horn of the hoof. Hoof samples for culture (using special footrot kit in field) Usually affects a number of sheep in the flock. a dark lead pencil to identify smears is also satisfactory. The lesion resolves quickly if sheep are moved to dry pasture or if footbathed in formalin or ZnSO4. Smears of affected feet lesions for D nodosus detection iii. Under certain conditions isolation and demonstration of virulent strains of D nodosus in the laboratory can be performed using a special footrot kit for sample collection. Use of a diamond pencil for numbering is recommended. However.nsw. Can cause marked lameness. particularly in rams and heavy sheep. including host resistance and stocking rate. laboratory culture for virulence testing may be necessary. (http://www. This exudate should abound in D. clinical examination of a sufficient number of sheep in the flock.au/reader/sheepfootrot/a0956. In some flocks. with a corresponding description of the animal/lesion on the specimen advice sheet. Hoof samples for culture (using special footrot kit in field) CLASSIFICATION OF DISEASE BASED ON CLINICAL EXAMINATION For full discussion of footrot classification (including colour photographs) refer to AgFact . of all age groups. and differentiate from virulent footrot. NB • Smears should be made from 5-10 affected animals.agric. the Regional Veterinary Laboratory should be contacted. When preparing the smear. In ovine footrot.Footrot in sheep and goats. avoid drawing blood.gov. • All slides should be numbered clearly and serially. If slides with frosted glass ends are available. ii. additional factors will also influence disease spread and expression. ii. but in some cases. as this renders examination of the smear more difficult. Diagnosis History. VETERINARY LABORATORY MANUAL 54 .distinct band of active necrosis can be observed along the healthy/ infected border. Cannot be differentiated from early virulent footrot. A small percentage may have some underrunning of the soft horn of the heel. This area is usually a couple of millimetres wide and is characterised by a greyish to yellow grey exudate. See Virulent Footrot. Seen in sheep grazing wet pastures under mild weather conditions. a second examination 10 to 14 days later will be necessary to check for progression of the disease. This may take a little while to download. Examination of sheep after a period of spread (wet pasture conditions/mild temperature) is more reliable to assess flock status. BENIGN FOOTROT IN SHEEP Caused by infection with benign strains of D nodosus. since legal action may result from the examination findings. B. The disease is best diagnosed in the field. Specimens required i. Culture of lesions and subsequent testing of D nodosus isolates may assist the field veterinarian to differentiate between benign and early virulent footrot.pdf?). • The differential diagnosis of footrot and foot abscess in sheep is best made on clinical examination. In such cases. with laboratory confirmation where necessary. Smears may be prepared from the moist necrotic interdigital skin if there is no underrunning of the heel or sole. Smears should be prepared by spreading a small amount of exudate thinly on a microscope slide (using a swab stick. nodosus and is the material of choice for diagnostic purposes. Bacteriology. Affected sheep show necrosis and inflammation of the interdigital skin. involving part or all of the soft horn of the axial wall. match or point of a knife).

refer (i) to (iv) below • In these circumstances a negative gelatin gel test may be used to support a field diagnosis of benign footrot iii. Disease is also considered virulent if: • Chronic cases of severe underrunning (score 4) are present irrespective of prevalence. Based on the examination of at least 100 sheep. • Score 4 lesions are present in obviously recent cases.none have foot scores greater than score 2. • The veterinarian is confident of a diagnosis of virulent footrot and can justify this (with adequate records). it may require examination of more sheep to detect a sufficient number of affected sheep (score 2 or greater) to assist in the diagnosis. virulent infections will persist at or about the same level but benign cases will have regressed without treatment VETERINARY LABORATORY MANUAL 55 .25% or more sheep have D. irrespective of the flock prevalence. particularly in areas of the State where the microclimate is unfavourable for full disease expression.no recent treatments have been undertaken and . Disease is probably benign but reinspection is advisable if: • After eradication programs on properties a very low prevalence of underrunning is seen in affected sheep the following spring. or. ii. Disease Classification Virulent footrot May be virulent or benign.FOOTROT SCORING SYSTEM Score Description 0 normal 1 non-specific inflammation and/or necrosis of the interdigital skin (IDS) 2 inflammation of the IDS which is due to infection with footrot 3 any lesion in any claw which results in underrunning of the soft horn of the heels or sole 4 underrunning of any hard horn of the claw In all circumstances where lameness is occurring. in individual mobs subsequent to eradication or in recently purchased mobs of sheep. This may be: • Partial expression due to recent virulent infection • Inadequate environment • Over expression of benign footrot in a very favourable environment • On reinspection 2-3 weeks later. Caution in footrot diagnosis is indicated and reinspection necessary if a proportion have score 3 lesions. where conditions have been conducive for footrot and no treatments undertaken since the eradication program. this will involve the examination of at least 100 sheep. the prevalence of score 4 lesions relative to the number of footrot-affected sheep can then be used to make the following classifications: % of affected sheep with score 4 lesions 1% or more < 1% i. recent virulent infections will have progressed under favourable conditions • If the environment is inadequate. In most mobs.at least 100 adult sheep are examined and . iv. nodosus infections and . In some mobs. it is recommended that sufficient sheep be examined to ensure that virulent footrot is not present. Virulent footrot can be excluded if: • Other identifiable causes of lameness are demonstrated and there is no evidence of D. nodosus in any sheep examined (minimum of 100) • After the mob has been through conditions favourable for the expression of footrot and . or. A negative gelatin gel test may be used to support a field diagnosis of benign footrot.

A negative gelatin gel test may be used to support a field diagnosis of benign footrot. ROLE OF THE LABORATORY IN FOOTROT DIAGNOSIS A laboratory test must not be used on its own to establish a diagnosis of virulent footrot. Where concern exists on the status of the disease on the property, the producer may elect, at the producer's cost, to utilise laboratory testing as an aid to the flock diagnosis. Culture and virulence testing of D. nodosus is expensive and labour intensive, with cultures requiring 8-18 days to isolate the organism in sufficient purity for characterisation. The gelatin gel test is currently used as the standard laboratory test to evaluate isolates as being benign or virulent (nonbenign). The elastase test is available but is not routinely used for estimating isolate virulence. As a minimum, it is anticipated that at least 100 sheep would have been examined in the mob and the lesions in individual sheep recorded, before specimens are collected for laboratory testing. Suitable samples from a minimum of 5 representative affected animals should be forwarded together with an adequate description of lesions in the sampled group. This will allow a minimum of 5 (up to a maximum of 20) isolates to be examined in the laboratory. The following information must be included in the specimen advice form: • number of sheep examined • foot scores of affected sheep • pasture conditions and rainfall over the past 3-4 weeks • previous footrot history of the flock • history of recent introductions • history of recent treatments • age, breed and sex of the sheep Laboratory-based tests should be interpreted in relation to clinical findings. In general, a positive test result is meaningful, but a negative test result is not a guarantee of footrot freedom. A negative gelatin gel test may be used only to support a field diagnosis of benign footrot. Refer to the booklets:
VETERINARY LABORATORY MANUAL

‘Footrot - technical information manual’, published by Coopers Animal Health Australia (for use with the NSW Footrot Strategic Plan) 2nd edn, 1990, edited by R.Walker, SFVO Wagga. ‘Footrot Strategic Plan 1991’, published by NSW Farmers, NSW Department of Primary Industries, Rural Lands Protection Boards, Stock and Station Agents Association and University of Sydney. ‘Proceedings of Dept of Animal Health Footrot Workshop - Gundagai March 1992’, by J.R. Egerton, University of Sydney.

FOOTROT IN GOATS AND CATTLE
In goats, the clinical signs are not necessarily indicative of the virulence of the organism. Benign sheep strains of D nodosus can cause severe underrunning and lameness in goats; conversely virulent sheep strains may only cause interdigital dermatitis in some goats. Where sheep and goats are run together and spread periods have occurred, then presence of footrot in the goats only is suggestive of a benign strain. Virulence of D nodosus isolates in goats will otherwise require laboratory testing of those isolates. With rare exceptions, cattle isolates of D nodosus to date have proven to be benign for sheep. However it has been shown that a virulent strain for sheep can be harboured by cattle with lesions of foot underrunning and/or discharge. Diagnosis See’Virulent footrot in sheep. Specimens required See’Virulent footrot in sheep.

FUNGAL INFECTIONS (OTHER THAN MYCOTOXICOSES)
Care should be taken in handling specimens from suspect cases as a number of fungal infections are communicable to man. Diagnosis Clinical findings, histopathology.

mycology

and

Specimens required Skin conditions i. Skin biopsy taken from the margin of an active lesion submitted for mycology* and histopathology is the best specimen.
56

ii.

Hair and deep skin scrapings from the peripheral areas of active lesions, submitted dry, for mycology*.

syndrome/diseases_of_livestock/dpi-sampcoll-prot.pdf (in order of preference) Diagnosis of the disease i. 20 to 25 hairs, with roots attached, from the distal end of the tail. ii. Lymphoid tissue and/or whole brain in buffered formalin for histopathology. Heterozygote detection i. 20 to 25 hairs, with roots attached, from the distal end of the tail.

Systemic conditions i. Sections of any organs showing lesions submitted for mycology*. ii. Sections of liver, spleen and lung submitted for mycology*. iii. Sections of affected organs and liver, kidney and spleen in buffered formalin for histopathology. *Samples for fungal culture should not be chilled or frozen, as temperatures of less o than 15 C can be detrimental to fungal survival. Culture of fungi may take up to 3 weeks.

GENETIC DISEASES
The majority of inherited diseases recognized in cattle are autosomal recessive. Due to the practice of “linebreeding”, some recessive defects may become relatively common. In addition, undue emphasis upon estimated breeding values (EBVs) to maximize production may lead to high levels of inbreeding and an inevitable increase in prevalence of recessive defects. In contrast, inherited defects that are autosomal dominant are most likely limited to a single herd, or rarely, to a few associated herds. Haemophilia A in Herefords is an example of an X-linked defect, where the clinical disease is most common in males. Potentially there will also be Y-linked defects and also those resulting from mutations in mitochondrial DNA. Autosomal recessive defects are expressed in male and female progeny of clinicallynormal, heterozygous parents. One quarter of the progeny of heterozygotes will be affected, half will be heterozygotes like the parents, and the remaining quarter will be homozygous wild-type (normal) - that is, they did not inherit the disease-causing mutation. Registered animals of most cattle breeds have been derived from a limited gene pool. Within modern breeds, herd books are generally closed. Consequently, most recessive defects are usually “breedspecific” and are caused by founder effects relating to a single mutation. There are exceptions where multiple mutations cause the same clinical disease within and across breeds. An affected individual may be heterozygous for two distinct mutations. The term ‘compound heterozygote’ is used to describe these animals. An example would be a Brahman calf affected with generalised glycogenosis if it inherited the exon7 mutation from one parent and the
57

GENERALIZED GLYCOGENOSIS
Syn: Pompe’s disease (Beef Shorthorn and Brahman cattle) The most common clinical presentation is of progressive muscular weakness leading to collapse and inability to rise, usually, but not exclusively, after weaning. There are no pathognomonic gross necropsy findings. PAS positive cytoplasmic vacuolation of peripheral lymphocytes is indicative of the disease. Similarly, light microscopic findings of PAS positive cytoplasmic vacuolation of lymphoid tissue, liver, kidney or neurons are indicative of generalised glycogenosis. There are three known “lethal” mutations in the acidic α-glucosidase gene in cattle, two in Brahmans (commonly known as E7 and E13) and a third in Shorthorns (commonly known as E18). The E7 mutation is the most common, with the E13 mutation restricted to descendants of one American import. Submitters are requested to specify breed of the subject. Submitters are advised to determine (via the Australian Brahman Breeders’ Association) if a Brahman subject is a descendant of the founder for the rare E13 mutation. Diagnosis Clinical signs and history, histopathology, confirm by DNA analysis. Specimens required See http://www.dpi.nsw.gov.au/agriculture/vetm anual/specimens-by-disease-

VETERINARY LABORATORY MANUAL

exon13 mutation from the other parent. Similarly, different mutations in the αmannosidase gene cause α-mannosidosis in Angus and Galloways. Consequently, an affected Angus/Galloway calf could be heterozygous for both of the mutations. In like manner, a Poll Hereford cross Poll Shorthorn calf may be affected with maple syrup urine disease if it inherits the “breedspecific” mutations from the parents. DNA tests are available for diagnosis of the following genetic diseases, or for detection of heterozygotes. With CVM in Holsteins Breed of Cattle Angus Murray Grey Galloway Salers Holstein/Friesian

and Dwarfism in Dexters, samples should be submitted direct to an external reference laboratory – addresses provided. With DNA-based assays, hair root samples are preferred as they avoid potential problems created by haemopoietic chimerism, which can occur when using tests based on analysis of DNA from cellular elements of blood. Bovine co-twins commonly share haemopoietic stem cells, resulting in DNA in cellular elements of blood containing nucleotide variants from both twins that may be different to the DNA variants found in gametes from an individual co-twin.

Poll Hereford Limousin Beef Shorthorn Brahman Dexter

Disease α-Mannosidosis α-Mannosidosis α-Mannosidosis ß-Mannosidosis Citrullinaemia Factor XI deficiency DUMPS Bovine Leucocyte Adhesion Deficiency (BLAD) Complex Vertebral Malformation (CVM)* Maple Syrup Urine Disease Inherited Congenital Myoclonus Protoporphyria Generalised glycogenosis (Pompes Disease) Generalised glycogenosis (Pompe's Disease) Chondroplasia (dwarfism) / Congenital lethal chondrodysplasia (Dexter bulldog syndrome)**

* Consign samples direct to Dr. Van Haeringen, Laboratorium B.V. PO Box 408, 6700 AK, Wageningen, The Netherlands. ** Consign samples direct to Reprogen, Faculty of Veterinary Science, University of Sydney, PMB 3, Camden, New South Wales 2570 See Specimens (by disease) for details of specimen collection for diagnosis of a specific disease.

GOITRE
Diagnosis Clinical findings, histopathology. Specimens required Thyroid glands submitted formalin for histopathology.

in

buffered

History of access to grain and demonstration of excess grain in ingesta with a pH of the ruminal content of less than 5. The rumen pH must be taken in the field and is only reliable if taken shortly after death. Elimination of bloat, enterotoxaemia as possible causes. Specimens required i. Serum or eye fluid for D-lactate. ii. Sections of wall of forestomachs in buffered formalin for histopathology. Units mM/L Normal < 0.4 Acidosis >1.0

GRAIN POISONING
Diagnosis Interpretation of D-lactate concentration Analyte Sample D-lactate Serum, aqueous humour There is no specific diagnostic test available for grain poisoning. Specimens should be taken to eliminate other possible causes, e.g. enterotoxaemia.

GRASS TETANY
See Hypomagnesaemia

VETERINARY LABORATORY MANUAL

58

toxic). tick fever. and mask to prevent inhalation of aerosols. parasitic. bacterial meningoencephalitis. Blood in sodium fluoride for elimination of hypoglycaemia. muscle fasciculations) and death. Range of samples for bacteriology. Veterinarians should take biosecurity precautions when performing post mortem examinations of horses. Hendravirus is a member of the paramyxovirus family of viruses. fatal respiratory disease that killed 14 horses. gasserian ganglion (trigeminal nerve ganglion). HELIOTROPE POISONING See: Pyrrolizidine alkaloidosis HENDRAVIRUS INFECTION Syn: Equine morbillivirus infection Hendravirus infection was first recognised in 1994 in Australia. liver. One human death occurred 14 months after exposure at post mortem examination of an affected horse. Hendravirus infection is notifiable. brain.g. SI content and a composite of visceral organs (liver. Japanese encephalitis. blood-tinged foamy discharge from mouth and nares). Specimens required i. iii. when it caused an outbreak of acute. cervical spinal cord. causing serious illness and sometimes death in humans. other neonatal diseases e. Diagnosis Histopathology. Consider exotic diseases in differential diagnosis of nervous disease: Aujeszky's disease. where possible. Fresh. leptospirosis. and one non-fatal human infection. All known cases have occurred in Queensland and coincided with the flying fox breeding season in the later months of the year. and is closely related to Nipahvirus. v. respiratory distress (dyspnoea. nutritional. paired virus serology. hypoglycaemia. spleen) collected aseptically into sterile Macartney bottles for virus isolation. HAEMATURIA See also: Enzootic haematuria of cattle. Suitable personal protective equipment includes gloves. During this outbreak the horse-trainer also died and there was a non-fatal infection of another person closely involved with the sick horses. virus isolation. overalls. samples of other visceral organs for histopathology. Clinical signs include pyrexia. To December 2004. sometimes neurological changes (headpressing. copper poisoning' HAEMOPHILIA See: Factor VIII deficiency (Hereford cattle) and Factor XI deficiency (Friesian cattle)' HEEL ABSCESS . bracken fern poisoning. differentiate from other causes of runting (bacterial. Nipah virus has flying foxes as its natural reservoir. Fees for tests undertaken to confirm or exclude a diagnosis of Hendravirus infection and to establish an alternative diagnosis are paid by NSW Department of Primary Industries. Flying foxes (fruit bats) are the subclinical reservoir of Hendravirus.HAEMAGGLUTINATING ENCEPHALOMYELITIS VIRUS (HEV) INFECTION OF PIGS Syn: Vomiting and wasting disease HEV is responsible for two overlapping diseases of young pigs: • Encephalomyelitis (with nervous signs) of pigs < 10 days old • Vomiting and wasting disease (with wasting and failure to thrive) of pigs from 7-21 days old. ii. Caution Hendravirus is a zoonotic agent. chilled samples of brain. Fixed brain. four more sporadic outbreaks reported have involved the death of six horses and one human.OVINE See: Foot abscess in sheep' VETERINARY LABORATORY MANUAL 59 . Call customer service (1800 675 623) for advice. poisoning-plant' HAEMOGLOBINURIA See also: Hypophosphataemia. kidney. which caused a major outbreak of acute fatal respiratory disease in pigs and humans (and dogs) in Malaysia in 1999. Teschen disease. small intestine. kidney. cervical spinal cord. iv.g. Differentiate from enterovirus encephalomyelitis. Serum samples from acute and convalescent phases (14 days apart). for differential diagnosis e. Like Hendravirus. lung. Colic was the initial sign in two recent cases.

Diagnosis Clinical signs. HYPOCALCAEMIA Diagnosis Clinical signs. Samples of affected tissues and swabs in Amies charcoal transport medium for bacteriology. free of cells and haemolysis. whole affected joint unopened (arthritis) or synovial fluid and synovial membrane (arthritis).au/health/3892. abortion. ovis/ H. Specimens required i. Best at papule/vesicle stage.complication or differential diagnosis of hypomagnesaemia. liver. Chilled lung for virology. lung. including histopathology. submitted chilled for calcium and magnesium estimation. iii. virology.g. Specimens required i. HEPATOSIS DIETETICA A disease of pigs. vesicles and ulcers which heal in 7-10 days (similar to cows with IPV). dysuria. fibrinopurulent leptomeningitis and ependymitis. e. Portions of affected tissues fixed in buffered formalin for histopathology.Post mortem findings include massive pulmonary congestion and oedema with airways filled with foamy. Further information at the Queensland Department of Primary Industries' website at http://www2. findings. and myocardium in buffered formalin for histopathology. gangrenous mastitis. milk and affected gland (mastitis). pneumonia. septicaemia. hypophosphataemia. portion of small intestine with mesentery attached. affected epididymis (epididymitis). straining. responsive to vitamin E/selenium therapy. Specimens required i. ii. lung (pneumonia). staggering and possibly mild ascites before sudden death. whole aborted foetus and membranes (abortion).gov. vaginal/uterine swabs and exudate (reproductive). ii. sometimes bloodstained fluid. Specimens required i. ketosis. 10 ml vacuum tubes of clotted and LiHep blood for virology. somnus from affected tissues NB The organism may be a commensal. skeletal muscle. polyarthritis. Swabs of vaginal/preputial lesion secretion in PBGS for virus isolation. HISTOPHILUS OVIS/ HAEMOPHILUS SOMNUS INFECTIONS Histophilus ovis and Haemophilus somnus are closely related organisms sometimes associated with the following conditions: Sheep Epididymitis. response to therapy. Fixed lung for histopathology. carried in the urogenital or upper respiratory tract of normal animals. Liver. pneumonia and nephritis. HERPESVIRUS VULVOVAGINITIS/BALAN0POSTHITI S IN GOATS See also: Infectious bovine rhinotracheitis (IBR). Specimens required i. abscess swab and/or abscess content (abscessation). as virus disappears with ulcer formation. (Overseas septicaemia. preputial/ vaginal swelling with papules. Diagnosis Clinical signs of dullness. mastitis). VETERINARY LABORATORY MANUAL 60 . serum or eye fluid calcium level .dpi. and less commonly. ii. Isolation of H.qld. At least 2 ml of serum.html . Virus isolation and virus serology. post mortem histopathology. Cattle (in Australia) Purulent vaginitis and endometritis. tendonitis. anorexia. infectious pustular vulvo-vaginitis (IPV) Diagnosis Clinical signs and history of mating season. polyarthritis and abscessation in lambs. spleen (septicaemia). thromboembolic meningoencephalitis. Diagnosis Clinical findings and pathology. Serum sample submitted chilled for virus serology. brain (CNS). kidney.

1 ml of urine for magnesium estimation.4 mmol/L Serum < 2. VETERINARY LABORATORY MANUAL • • Parasitism.7 mmol/L > 0. Interpretation in cattle and sheep Calcium Serum > 2. Specimens required i. clinical signs. response to therapy.5 1 ml of aqueous humor. for magnesium estimation. Specific bacterial diseases e. Cases of ill thrift associated with anaemia or scouring should be investigated under the syndromes of 'anaemia' or 'scouring'. NB Serum phosphorus levels are of little value if the sample is haemolysed or contaminated. submitted chilled for calcium and magnesium estimation. ii. HYPOMAGNESAEMIA Syn: Grass tetany.45μ membrane filter) to minimise cellular contamination. clinical signs.25 mmol/L Normal Aqueous > 1. Alternatively.g. the commonest cause. Other causes of ill thrift not necessarily associated with anaemia and/or scouring include: • Malnutrition. iii. lactation tetany.45μ membrane filter) to minimise cellular contamination. free of blood and cells. EBL in cattle.ii. caseous lymphadenitis.6 mmol/L > 2 μmol/mosmol HYPOPHOSPHATAEMIA Diagnosis History. At least 2 ml of serum.5-1 ml of aqueous humor. Specimens required Interpretation in cattle and sheep Normal Deficient i. response to therapy. • Nutritional deficiencies of copper cobalt. From cadavers. scouring' Ill thrift is a vaguely defined condition with a variety of causes. including helminthiasis. submitted chilled or frozen for phosphorus estimation. Do not send serum on the clot. filter aqueous humour (0. serum. filter aqueous humour (0. Johne's disease. CAE in goats. Ideally. free of blood and cells for calcium estimation. Serum Serum • • • Phosphorus > 1.1 mmol/L Deficient Aqueous < 1. Diagnosis 61 . clinical signs. area information. Specimens required There is no routine laboratory test for blood glucose concentrations. phosphorus or selenium. pestivirus infections in ruminants. sarcosporidiosis and chronic infectious diseases Viral: EIA in horses. milk tetany of calves Diagnosis Interpretation in cattle and sheep Normal Serum Aqueous Urine Magnesium > 0. free of cells and haemolysis.3 mmol/L < 1. differentiation from ketosis. At least 2 ml of serum free from cells. After effects of systemic or chronic conditions including pyrrolizidine alkaloidosis. From cadavers.1 mmol/L HYPOGLYCAEMIA Diagnosis History. Ideally. fascioliasis and coccidiosis. eye fluid or urinary magnesium concentration. 0. Eperythrozoonosis. blood or serum inorganic phosphate concentration. History.0 mmol/L ILL THRIFT See also: Anaemia'. 0. complications of hypocalcaemia.

Specimens required Respiratory and ocular disease i. abortion or perinatal loss. most frequent being fractures of the acetabulum. and less commonly. i.Based on clinical findings. including brain for virus isolation and histopathology.dpi. cardiac and skeletal muscle. Swabs from nasal cavities and conjunctivae in phosphate buffered gelatin saline (PBGS). genetic INFECTIOUS BOVINE RHINOTRACHEITIS (IBR) AND INFECTIOUS PUSTULAR VULVO VAGINITIS (IPV) Diagnosis Virus isolation and antibody rise between acute and convalescent serum samples. Swabs of vaginal mucus in PBGS. 20 to 25 hairs. virological and/or parasitological examination. sera and/or portions of body organs including the above. small and large intestine or other organs depending on gross pathological indication of aetiology at field necropsy. semen examination. INFERTILITY IN CATTLE. The majority of affected calves have hip lesions. INHERITED CONGENITAL MYOCLONUS (Poll Herefords) Stimulus-responsive myoclonic spasms in bright and alert calves is the striking clinical feature of this disease. ii. A detailed history should be provided to allow consideration of other causes. kidney. Encephalitic form i. at joining due to fertilisation failure (male or female infertility). PIGS See also: Abortion (general). iii. Detailed investigations should involve prior consultation with the laboratory. submitted chilled for virus serology.pdf Diagnosis of the disease and heterozygote detection i. Live unthrifty animal submitted for post-mortem examination. The first step is to establish the stage in the reproductive cycle where the problem occurs. laboratory examination to eliminate specific causes and response to treatment. epididymitis. with roots attached. 62 VETERINARY LABORATORY MANUAL . Specimens as required for encephalitis. Abortion i. embryonic loss. submitted fresh for bacteriological. Blood samples as required to eliminate specific deficiencies of selenium. INHERITED DEFECTS See congenital diseases' abnormalities. ii.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot. Acute and convalescent serum samples (collected 3 6 weeks apart). congenital abnormalities. submitted chilled for virus serology. v. Specimens required Those appropriate to the condition or conditions suspected.nsw. clinical signs. No consistent histological findings. If indicated. ii. copper deficiency. Diagnosis History. Specimens required See http://www.gov. from the distal end of the tail. The conditions listed should operate on a flock basis so that specimens should be submitted from a number of animals in the flock. of liver. Faecal samples from 10 to 20 animals for parasitology. Acute and convalescent serum samples (collected 3 6 weeks apart). Sections fixed in buffered formalin.e. The range of specimens to be submitted could include: i. iv. Vulvo-vaginitis: i. Specimens as required for bovine abortion. phosphorus and copper. confirmed by DNA analysis. fractures of the head of the femur. trichomoniasis of cattle Diagnosis Must be based on field investigations supported by the laboratory where necessary. SHEEP. Specimens required Specimens should be submitted to eliminate some of the possible causes only after malnutrition has been eliminated as a contributing factor.

goat. submitted chilled for selenium analysis. lice. examined in the field. It is of value only in those animals with moderate histological lesions (including all clinical cases). Other specimens appropriate to the disease condition suspected see Copper poisoning. Specific causes diagnosed on the basis of laboratory examination. The C strain can be cultured by either conventional (solid media) culture or by radiometric broth (Bactec) culture. deer and alpaca infections are attributed to C strains. Sections of liver. Specimens required In cases of dermatitis or skin irritation where mites cannot be seen at field examination. Specimens required i. Specimens required i. NB Wool must be clipped as close to the skin as possible before taking a scraping (see Parasites . conventional culture is usually slower than Bactec culture. Common causes include plant poisonings. The majority of cattle. history. but its specificity is extremely high. and a longer ITCHMITE IN SHEEP Diagnosis Demonstration of Psorergates ovis mites in deep skin scrapings from suspected cases. 10 ml of heparinized blood from sow(s). while sheep infections are predominantly due to S strains. Faecal and tissue culture Faecal and tissue culture procedures depend on whether the cattle (C) or sheep (S) strain is likely to be involved. AGID An AGID is available for goats and sheep this has a low sensitivity in identifying infected animals. ELISA positive goats can be further tested by AGID to identify those infected animals with most advanced lesions. Differentiation from other causes of fleece derangement e. Plant poisoning. kidney and spleen. tender fleeces. submitted in buffered formalin for histopathology.IRON TOXICITY SYNDROME IN PIGLETS Diagnosis Clinical history of sudden death of 0-7 day piglets within 1 12 hours of giving iron injection. Eperythrozoonosis. In the live animal. The S strain can only be reliably cultured by radiometric (Bactec) culture. a positive test is indicative of current histological lesions of the disease.g. and should be restricted to clinical cases. A skin biopsy from the affected area. Serology. JOHNE'S DISEASE Syn: Paratuberculosis Diagnosis History. Cattle or goats reared with infected sheep may be infected with S strain. paratuberculosis infection and disease. ii. the following samples may be sent to the laboratory: i. For the C strain. goat. clinical signs. leptospirosis and eperythrozoonosis. Piglet for post mortem examination and differential diagnosis. gross pathology and histopathology. JAUNDICE Jaundice is a clinical sign which may arise as a result of a haemolytic crisis or hepatic failure.External). chronic copper poisoning. submitted in buffered formalin (may assist in a differential diagnosis). submitted with the blade enclosed in a secure sample container. Absorbed ELISA testing is the most sensitive serological method for diagnosis and certification for M. Isolation of Mycobacterium paratuberculosis from faeces or tissues. ii. but is less expensive. alpaca and deer sera. C strain infection in sheep is relatively uncommon. Pyrrolizidine alkaloidosis. Diagnosis Clinical signs. Demonstration of typical organisms in faecal smears and tissues. It also has a very high specificity. Skin scrapings collected using a scalpel blade dipped in paraffin oil. serum chilled for enzymology. selenium deficiency in sow. VETERINARY LABORATORY MANUAL 63 . Absorbed ELISA Absorbed ELISA are routinely available for Johne's Disease testing of cattle. Leptospirosis. iii. or vomiting/ diarrhoea/ dyspnoea/ coma and death up to 3 days. ii. Smears of faeces and tissues Smears of faeces and tissues are of limited sensitivity in subclinical animals.

paratuberculosis confirmed Total time if growth detected and not M. The following table outlines expected time differences between the two strains in current cultural procedures: S strain Bactec 5-7 d 12 wks 1-4 wks Up to 10 wks 13 wks 10-23 wks 12-23 wks If the default procedure is not suitable (i. paratuberculosis are targeted in the confirmation procedure for maximum specificity. IS1311 PCR and REA) can reduce the time to complete confirmatory testing procedures. deer: Goats. longer incubation. the required procedure must be advised on the Specimen Submission Form. Examination for the second characteristic of M. at week 8). Together. Two separate characteristics of M. paratuberculosis from culture for C strains is the second part of the Bactec procedure i. paratuberculosis. these can identify the presence of M. a confirmatory phase. and. to reach a conclusion of culture positive for M. The current default procedures (based on consumer demand) apply to culture submissions from cattle. as this optimizes the confirmation rate. and takes up to 10 weeks. paratuberculosis as part of the confirmatory process from Bactec media is also commenced when growth reaches a high level. incubation to week 10 may be required to reach a satisfactory level for confirmation. subculture for 64 VETERINARY LABORATORY MANUAL . goats. Different solid media are used for subculture of the C and S strain. This normally requires a sample of growth to be subcultured to solid medium for evidence of typical colonies with mycobactin dependency. Confirmation conventional equivalent to confirmatory of M. both characteristics need be met on only one tissue for a conclusion of culture positive status for that animal. paratuberculosis Confirmation of M.e. it does so less efficiently than the preferred C strain medium. S strain does not grow on the solid C strain medium.g. the alternate procedure is required). if growth occurs. In addition. When multiple tissues are cultured from an individual. The culture process is undertaken in batches. As an alternative to subculture to demonstrate mycobactin dependency. where OJD is suspected and a non-ovine species is being tested by culture. alpaca and deer: Species Cattle. The amount of M.incubation period is used compared to the C strain. different subculture medium) can be undertaken. paratuberculosis C strain Conventional Bactec 5-7 d 5-7 d 20 wks 8 wks* 1-4 wks (if reqd) 1-4 wks Up to 10 wks 21 wks 12-26 wks 12-24 wks Up to 10 wks 9 wks 10-21 wks 12-21 wks * If growth is detected late (e. Samples of Bactec growth are collected and then processed by PCR and REA in batches. paratuberculosis in the sample correlates with the rate of growth (higher numbers reduce the time delay until growth is detected). alpacas: Method Bactec Conventional be clear on the specimen advice form so that procedures specific to the S strain (i. The first characteristic is measured in a molecular assay involving a polymerase chain reaction (PCR) and restriction endonuclease analysis (REA) that target the IS900 gene sequence. testing for a second specific molecular target (e. and yet retain maximal test specificity. This in turn modifies the time for culture to be completed.g. While solid media that supports the S strain can be used to cultivate the C strain. Culture confirmation of M. In contrast.e. followed by an incubation phase. this must Method Decontamination Incubation Confirmation by PCR/REA Confirmation by subculture (mycobactin dependency test) Total time if no growth detected Total time if growth detected and M. paratuberculosis from Bactec growth is undertaken when growth reaches a high level.e. and requires an initial decontamination phase over 5 working days. for greatest efficiency. paratuberculosis DNA with very high specificity.

choose additional sheep to allow for animals from which no sample can be collected (allow 55 sheep for each pool of 50 pellets). This should be borne in mind when completing the Specimen Submission Form for samples from herds without prior confirmation of Johne's disease and where conventional culture is requested. *Note: Pooled faecal culture procedures for cattle and goats are currently under development. with false positives and false negatives occurring frequently. lymph node enlargement). in buffered formalin for histopathology. In goats and sheep. goats) and/or AGID (goats. chilled portions of intestine and intestinal lymph nodes in separate sterile containers for bacteriology. • Serial faecal culture . It is restricted for export/certification testing only. faeces at slaughter can be useful to assess the risk of shedding.in sheep one pellet from each of (up to) 10 sheep and submit from these same animals on each of three occasions 10-14 days apart. NB: Allergic tests and serological tests such as the CFT are unreliable in the diagnosis of the disease. The best samples are usually those with obvious lesions (gut thickening. Fresh. do not forward entire intestinal tract for laboratory examination without prior consultation. In flock sampling. Cattle: Intestinal portions of 5 cm x 10 cm • proximal ileum (30-45 cm anterior to ICV) • terminal ileum (11-20 cm anterior to ICV) • ileocaecal valve (ICV) and attached segment of ileum • proximal colon • ileal LN • ileocaecal LN Goats and alpaca: Intestinal portions of 5 cm in length • jejunum • proximal and terminal ileum (1 m apart) • ileocaecal valve and attached segment of ileum • proximal colon • ileal LN • ileocaecal LN Sheep: • terminal ileum • ileal LN (caudal jejunal LN) ii. and not for routine diagnostic purposes. Sections of the above. submit only sufficient per pool to fill the 60 mL container. 65 VETERINARY LABORATORY MANUAL . In addition. Faeces. sheep). submitted chilled for bacteriology • Individual faecal culture . If pellets are l large.mycobactin dependency. Where this is not possible. Samples from the following sites per animal are considered ideal. Individual culling on the grounds of such tests is often impractical. Unless such samples are received soon after collection. autolytic changes can prevent useful examination.in sheep*. and are typical of those required for MAP certification.volume of > 5 g per animal • Pooled faecal culture . one pellet from each of (up to) 50 sheep per pool. It is restricted to use for risk assessment purposes to determine if high shedder animals are present. However. additional IS900 PCR and REA testing will be undertaken on growth on solid media typical of M. This relies on PCR and REA targeting the IS1311 gene sequence. can include a section of fixed liver (microgranulomata). Specimens required In the live animal: i. paratuberculosis in herds where Johne's disease has not previously been confirmed. Notes in regard to PFC submissions: • Do not take more than one pellet per animal • Samples must not overfill a 60 mL sterile plastic universal container. This will create a single pool of up to 30 samples. NB It is preferable that the above samples be taken in the field. include fixed caecum. The Johne's CFT is not routinely available. In cattle and goats. Strain typing Strain typing can be undertaken on Bactec growth if requested. Serum sample for ELISA test (cattle. In slaughtered animals: i. Direct PCR Direct PCR on pooled or individual faeces is available for sheep in limited circumstances. ii.

abattoir workers. Higher concentrations should be interpreted on the basis of clinical findings and histopathology. submitted chilled for toxicology. depression. and whole brain in buffered formalin for histopathology. KIKUYU POISONING A disease of cattle causing salivation. reticulum. biochemical analysis of tissues. Specimens required Live animal i. whereas serovar infections in non maintenance hosts are usually associated with sporadic outbreaks of disease. Dead animal i. infections by these specific serovars are likely to be endemic. abdominal pain and death associated with grazing on lush kikuyu pastures. particularly Northern Australia. The cause is unknown. Interpretation of lead concentrations (cattle and sheep) Analyte Sample Minimum Units Normal amount Lead EDTA blood 5ml µmol/L < 1. L pomona • Pigs L pomona.particularly in serological reactor animals with limited lesions. but such infections are rare in NSW. ii. ii. L tarassovi • Horses L pomona • Dogs L copenhageni Other serovars do affect domestic animals in Australia. hardjo L. copenhageni Maintenance Host Cattle Pigs Rats In the maintenance host. although mycotoxins have been suggested in some cases. rumen abomasum and kidney. Faecal lead concentrations below 10 mg/kg are considered non-significant. Higher levels could be significant. The most common manifestations of leptospirosis in animals are: • Abortion (all species) • Milk drop syndrome/ mastitis (cattle) • Pyrexia/Septicaemia (all species) • Icterus and/or haemoglobinuria (mainly dogs and calves) • Kidney lesions found at routine slaughter (pigs. ii. fungi. Sections of omasum. Sections of liver and kidney. L. submitted in buffered formalin for histopathology (for differential diagnosis). Specimens required i. clinical signs of central nervous system disturbance. dairy and pig farmers. tarassovi L.2 Lead Lead Kidney (Liver) Faeces 50g 100g mg/kg (wet wt) mg/kg (wet wt) <4 < 10 Possibly toxic 4-25 10-25 Toxic > 1. LEAD POISONING Diagnosis History of illness. There is no value at present in examining samples of kikuyu for toxins. pomona. LEPTOSPIROSIS Leptospirosis is transmissible to man and represents an occupational hazard to veterinarians. etc. In calves. Zoonotic infections are usually acquired from the maintenance host for each serovar: Serovar L. 50 g of kidney. Diagnosis History. in buffered formalin for histopathology. cattle) VETERINARY LABORATORY MANUAL 66 . clinical signs and histopathology (particularly of omasum and reticulum).2 > 25 > 25 Kidney lead concentrations below 4 mg/kg are considered non-significant. Sections of liver. The most commonly encountered serovars associated with clinical disease are: • Cattle L hardjo. thymic haemorrhages are a frequent finding at necropsy. kidney and small intestine. dehydration. 10 mL EDTA blood tube. depending on the source of lead. 100 g of faeces.

this is a doubling dilution series starting at 1:25 (L hardjo) or 1:50 (other Leptospires). Kidney lesions in slaughter animals i. Submission of 10 15 sera from animals in the same group as the affected individuals will give a more accurate assessment of the leptospirosis status of the herd. urine or milk is not undertaken. L pomona is more immunogenic than L hardjo. Serum samples from 10 15 affected and recovered cases and cohorts. • A convalescent serum sample collected 10 days or more after agalactia often shows a rise in titre to L. whereas in L. Mortalities following pyrexia and/or icterus/haemoglobinuria i. pericardial). In an endemically infected herd at any given time. except under specific arrangements with the receiving laboratory. 1 ml of 10% VETERINARY LABORATORY MANUAL neutral buffered formalin to 99 mL urine) added immediately after collection. chilled entire foetus with membranes (or chilled and fixed foetal tissues. 1 ml of 10% neutral buffered formalin to 99 mls urine) added immediately after collection. Interpretation of Leptospiral titres For the MAT. titres at the time of abortion may be equivocal and falling. In L pomona infections and L hardjo milk drop syndrome. in buffered formalin for histopathology. Indication of current infection can only be gained by demonstrating at least a four-fold (i. This involves field-inoculation into selective liquid media of mid-stream urine collected after diuretic treatment. submitted as outlined in 'Mastitis'. To determine recent infection. titres are given which reflect the standard Leptospiral dilution series for sera. ii. including serous fluids) for bacteriology and histopathology. some will be static. iii. Milk Drop Syndrome i. Sample of urine preserved with formalin (approx. For leptospiral MATs. Specimens required Abortion Samples as outlined in 'Abortion in Cattle' or 'Abortion in Swine'. so titres to L pomona are often 800 or more. from both affected and recovered cases. and/or by demonstration of organisms in kidney by histopathology. and fresh. It is worthwhile using a diuretic to obtain a sample with large numbers of organisms and minimal contaminants. In cattle. ii. an MAT titre to L hardjo of 25 or greater may be significant. following consultation with the Regional Veterinary Laboratory. Serum samples from 10 15 affected and recovered cases and cohorts. iv. and others will be falling. In certain cases. titres generally rise within 1 3 weeks of the appearance of clinical signs. hardjo. Confirmed by demonstration of leptospires via direct microscopic examination of urine or foetal fluids (thoracic. they may be less than 800. Portions of affected kidneys in buffered formalin for histopathology. submitted chilled for serology. hardjo milk drop syndrome. In cases of abortions in cattle due to L hardjo. 2 dilution) increase. 10 15 paired sera (taken 2 4 weeks apart) should be submitted to detect rising titres (along with the clinical history of the tested animals). from both affected and recovered cases. some titres will be rising. foetal fluids. Sample of urine preserved with formalin (approx. peritoneal. Icterus/haemoglobinuria Samples as for Pyrexia. Urine shedding of leptospires occurs after the haemoglobinuric phase. arrangements may be made to sample cows for culture. Titres following vaccination are generally low (less than 400) and do not persist. Whenever possible. but do not collect urine for microscopic examination from animals affected with haemoglobinuria. Sections of liver and kidney. Pyrexia i. Samples of milk for differential diagnosis of mastitis.e. L hardjo titres of 200 or more are generally significant. In general culture of tissues. including 10 15 sera from cohorts. submit serum from dam.• Mortalities following pyrexia and/or icterus/haemoglobinuria Diagnosis Serology (Microscopic Agglutination Test). 67 . submitted chilled for serology. for dark ground microscopy.

Faulty application of treatment should be considered where infestations are localised to one side or part of the sheep. Abortion Post mortem findings and bacteriology. The false negative rate is low in cattle (<2%). Diagnosis Examination of sites of. Diagnosis VETERINARY LABORATORY MANUAL . NB. serology. or where one mob or line is infested and other mobs are free of lice. Demonstration of resistance using an in-vitro laboratory test. Septicaemia Bacteriology and histopathology. Resistance may be suspected on a history of treatment failure after correct application of the correct amount of lousicide at the correct time. clinical examination. ii.LICE RESISTANCE TO INSECTICIDES IN SHEEP Resistance of sheep lice to pyrethroid insecticides is widespread in NSW. • Where sheep in long wool have been thoroughly treated with a pyrethroid and failure to control infestation is obvious within 6 weeks. liver. Specimens required Meningoencephalitis i. Necropsy findings. or. Brain in buffered formalin for histopathology. ELISA serology is available at EMAI for the detection of infection with both immature and adult fluke. kidney and spleen. Meningoencephalitis (circling disease) Clinical signs and brain pathology. they are of no value in acute and subacute infections. No functional organophosphate resistance has been detected. and degree of. kidney. Using the Institut Pourquier (France) antibody ELISA recently adopted (2004) by NSW DPI laboratories. ELISA serology is more sensitive than faecal egg counts for the detection of chronic fascioliasis in both cattle (30% more infected animals detected) and sheep (1520% more infected animals detected). NB Long wool application of pyrethroids may leave undesirable residues at shearing. particularly in the case of shower dipping failures with insecticides. submitted chilled for bacteriology. Faecal egg counts are only of value in chronic fascioliasis where eggs are being excreted (patent infection). A thorough examination of the dip operation and thoroughness of wetting is recommended. Abortion i. in buffered formalin for histopathology. Submission of live affected sheep would be required. it requires controlled temperature and humidity and is a laboratory based test at the Elizabeth Macarthur Agricultural Institute (contact Dr Garry Levot). ii. Specimens required Testing for OP resistance is not routinely available. Heart blood and sections of liver. which typically occurs in 68 LISTERIOSIS Listeriosis can be transmitted to man. titres appear 2-4 weeks (cattle) after infection and remain high for at least 20 weeks after tretment. Sections of liver. It is more productive to inspect a larger number of sheep rather than search a smaller number of sheep more thoroughly. infestation in 4-5 affected sheep by counting lice in 10 partings on each side of the infested sheep (each side examined in 2 lines of 5 partings). Specimens as required for abortion in the particular species (see Abortion). Septicaemia i. The test has not been validated for sheep under Australian conditions. response to treatment. spleen and lung. LIVER FLUKE INFECTION See also Parasites Internal Diagnosis Grazing history. spleen and kidney. either: • As breakdowns within 3-4 months of application of backline treatment in sheep treated off-shears or in short wool. Cerebrospinal fluid and sections of brain. ELISA serology is the only antemortem diagnostic method for the detection of acute and subacute fascioliasis (before fluke eggs are excreted). faecal egg count. submitted chilled for bacteriology.

69 VETERINARY LABORATORY MANUAL . It can affect the joints. ii. abortion and laminitis like signs can be shown by affected cattle and horses. • In animals where liver fluke has been detected in the herd or flock and treatment has been given in the previous 5 months. Faecal samples may be tested individually. Differential diagnosis: • Other causes of arthritis: Chlamydiosis. some horses may have skin hypersensitivity. The ELISA test is reported by the manufacturer to be suitable for individual and bulk vat milk samples. Serum samples from affected cattle and their cohorts for ELISA serology ii. or. domestic animals and wildlife and is known to occur in Europe. Chlamydial Infections.nsw. clinical signs and histopathology. Specimens required i. The organism may be more readily isolated from blood during the early stages of disease. Lyme disease usually presents as lameness associated with polyarthritis. There is clinical.57 (second edition 1999 revised 2003) ‘Liver fluke disease in sheep and cattle’: http://www. NB For interstate testing for import of ruminants to Western Australia. In animals. i. Sections of liver and kidney in buffered formalin for histopathology. specimens as required for nervous disorders.gov. Western Australia web site under Livestock movement at http://www. Diagnosis Clinical signs. Specimens required At necropsy. Asia and North America. The milk ELISA is a herd test and for statistical significance a minimum of 12-15 samples should be submitted. fascioliasis should be diagnosed in the field and laboratory specimens will not be required. Submitters should nominate how they wish the samples to be tested. Faeces for fluke egg counts in the following situations: • Where serum collection in cattle is not feasible. • Ephemeral Fever. or • Where paired faeces and serology is specifically sought for comparative study.au). (typically Borrelia burgdorferi is usually present in only low numbers in affected joints. joints are painful and swollen and there may be a low grade pyrexia. with inability to rise in severe cases. or pooled in groups of two (designed for sheep) or five samples (cattle). where it is primarily transmitted by Ixodid ticks. uveitis and CNS signs resembling encephalitis. histopathology reveals a fibrinoid polyarthritis). stiffness and myalgia. Chronic weight loss. Likely vectors here are Ixodes holocyclus or other Ixodes spp. serological and pathological evidence of a Lyme Diseaselike in cattle and man in eastern coastal Australia but the organism has not yet been isolated.NSW from autumn to early winter and in late spring. Mycoplasmosis). Submit separate faecal samples (at least 30 g of fresh faeces each) from at least 10 animals in the flock or herd. by arrangement. Serology can be undertaken for some animal species outside of the Department's laboratories. The test is currently (2004) undergoing validation by NSW DPI. If nervous symptoms are seen. Diagnosis History.gov. For more information.wa. which is fluke-free. and requires special media for isolation. heart and nervous system of man. There may also be lethargy.pdf LUPINOSIS A mycotoxicosis mainly associated with grazing on lupin stubble infected with Phomopsis leptostromiformis.agric. LYME DISEASE Lyme Disease is a multisystem disease caused by the spirochaetal bacterium Borrelia burgdorferi.9. individual samples from up to 30 animals in each consignment are required (further details are available at Department of Agriculture. in a screw capped jar. refer to the Agfact A0.au/reader/andiseases/a0-9-57web. mycoplasmal arthritis (See Arthritis.agric. Pooling reduces costs but sensitivity is compromised.

Individuals regularly handling potentially-affected bats (wildlife carers. VETERINARY LABORATORY MANUAL 70 . MANGE Diagnosis Clinical signs. brain. selenium. virology. in buffered formalin for histopathology. copper. pyrrolizidine alkaloidosis) and deficiencies of particular nutrients (e. pteropid lyssavirus Australian bat lyssavirus is closely related to classical rabies virus and causes similar progressive neurological disease in naturallyinfected bats and humans.au/internet/wcms/Publi shing. e. histopathology. post mortem findings and recovery of pathogenic clostridia from lesions. otherwise examination is worthless. aggressive or unusuallydocile bats are at high risk of being clinically-affected). In cases with polyarthritis. chronic conditions causing organ dysfunction (e. Whole bat (chilled) MALIGNANT OEDEMA Diagnosis History of mortality. Specimens required i. salivation) and paralysis. Material must be taken from affected or freshly dead animals.gov. People bitten or scratched by bats should consult a General Practitioner immediately and seek prophylactic vaccination. Affected tissues from affected animal or freshly dead animal. Sections of affected tissues.g. In acute cases. Australian bat lyssavirus infection is notifiable. chilled joint fluid and (in PM material) joint capsule is sought. At least three impression smears from lesions for bacteriology. Fixed tissues for histopathology: joint capsule. Malnutrition should also be considered as a predisposing cause of many conditions. Where disease is suspected. iii.g. The differential diagnosis should include parasitism. All bats should be handled with care to avoid bites and scratches. liver. Flying foxes (fruit bats) and insectivorous bats are affected.health. heart. shivering. Best diagnosed in the field because of rapid autolysis of samples. causing fatal infections in humans. demonstration of parasites in deep skin scrapings or skin biopsy. histopathology. veterinary pathologists) should be vaccinated against rabies. Diagnosis Clinical signs.csiro.au/. Further advice can be obtained from the Australian Government’s Department of Health and Aging website at http://www. The general public should be discouraged from rescuing or handling bats (paralysed. Clinical signs in bats include behavioural changes (aggression/docility. Further information on this disease can be obtained from the CSIRO website at http://www. kidney.nsf/Content/cda-pubs-otherbat_lyssa.g.htm. which are the reservoir of the virus. Fees for tests undertaken to confirm or exclude a diagnosis of lyssavirus infection are paid by NSW Department of Primary Industries. heparinised blood is suggested. especially for alternative diagnosis (Chlamydia) ii. Caution Lyssavirus is a zoonotic agent. Specimens required i. bats should be euthanased and sent to the laboratory for necropsy. plant poisoning in drought situations. iii. MALNUTRITION See also: Feeds and pasture analyses Malnutrition is the most common cause of ill thrift. pregnancy toxaemia. phosphorus). LYSSAVIRUS Syn: Australian bat lyssavirus. Specimens required i. Subclinical infection apparently occurs in bats.• Septicaemic and toxaemic conditions caused by other bacteria. submitted chilled for bacteriology. skin from tick bite site (in neutral buffered formalin). Chilled acute and convalescent serum. Culture may be arranged by consulting RVL staff at EMAI. peripheral nerve. infectious diseases. ii.

breed of the subject must be recorded) Diagnosis of the disease BETA-MANNOSIDOSIS (Salers cattle) Affected calves are usually born alive. Hereford and Poll Shorthorn calves) An inborn error in branched chain keto acid metabolism that results in a progressive neurological disease that culminates in death before 5 days of age. from the distal end of the tail. 20 to 25 hairs. but exhibit severe neurological abnormalities. Heterozygote detection i. Murray Grey and Galloway cattle) Affected Angus and Murray Grey calves either fail to survive the immediate postnatal period. Widespread vacuolation of white matter tracts in the brain and spinal cord is a common histological manifestation of MSUD in Hereford and Shorthorn calves. The phenotype in Galloways is more severe. from the distal end of the tail. incoordination. There are no pathognomonic gross findings at necropsy. ii. ii. breed of subject must be recorded) Diagnosis of the disease i. 20 to 25 hairs.pdf (in order of preference.dpi. Specimens required See http://www. and aggression.gov. with roots attached. At birth. affected calves are clinically normal. clinical signs. MAPLE SYRUP URINE DISEASE (MSUD) (Poll Hereford. histopathology. in buffered formalin for histopathology (in cases where the disease is suspected but cannot be confirmed from skin scrapings). progressive neurological disease characterised by tremors of the head.dpi.gov. Skin scrapings.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot. confirmed by DNA analysis or histopathology . ii. MANNOSIDOSIS ALPHA-MANNOSIDOSIS (Angus. but within 24 hours they develop progressive neurological dysfunction characterised by uncoordinated limb movements that prevent the calves from rising. ataxia.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot. they show severe.dpi. History. 20 to 25 hairs. head-swaying.gov. clinical signs. confirmed by DNA analysis or histopathology. These include weakness.pdf (in order of preference) Diagnosis of the disease i. Diagnosis History. and a poor sucking reflex.nsw.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot. Lymphoid tissue and/or whole brain in buffered formalin for histopathology.nsw.Cases are best diagnosed in the field from direct examination of multiple deep skin scrapings. splaying of the front legs. Diagnosis History. Specimens required See http://www. Specimens required See http://www.pdf (in order of preference.nsw. Skin biopsy from a recently affected area. Diagnosis VETERINARY LABORATORY MANUAL 71 . Specimens required i. with roots attached. with most affected cases aborted or stillborn. with roots attached. with roots attached. from the distal end of the tail. clinical signs. or if they do. Death intervenes within 5 days of birth. Between 48 and 72 hours after birth the calves collapse and typically are often found in lateral recumbency with opisthotonos. from the distal end of the tail. Heterozygote detection i. Lymphoid tissue and/or whole brain in buffered formalin for histopathology. 20 to 25 hairs. confirm by DNA analysis.

description of clinical signs and any recent changes in farm management or milking parlour management. resample case at next occurrence of clinical signs or at least three weeks post treatment. Samples should be kept on ice until arrival at the laboratory. Enterobacter spp and other coliforms may be present. Escherichia coli. Interpretation of results Staphylococcus aureus. with roots attached. Whole brain in buffered formalin for histopathology. additional information required is number of cows involved. Approximately 20 ml of foremilk collected aseptically from affected quarter into a sterile container. All recurrent cases should be cultured to check cause. Herd mastitis problem Evidence of subclinical mastitis (see above). 20 to 25 hairs. problems in milking parlour management or milking machine operation. elevated milk NAGase activity. clinical signs and bacteriology Subclinical mastitis Bacteriology. 20 to 25 hairs. from the distal end of the tail. Specimens required i. from the distal end of the tail. For clinical cases of mastitis this should include breed. • Discard the first few squirts before collecting sample. • Before sample is taken. previous mastitis history. Individual cow cell counts are available through Herd Recording. teat must be washed and dried thoroughly and teat end sterilised with 70% alcohol or undiluted teat dip solution. Streptococcus dysgalactiae and Streptococcus spp (environmental) and are the most common pathogens found in both clinical and subclinical mastitis. In mastitis cases occurring at or near calving. Streptococcus uberis. Subclinical mastitis problems require an intensive investigation of milking parlour operation. Incurable mastitis can be caused by Pseudomonas aeruginosa.000 averaged over a lactation. • All samples should be taken prior to antibiotic treatment. Mastitis due to Arcanobacterium pyogenes usually necessitates drying off affected quarter after treatment and reassessing damage to the quarter after subsequent calving. 72 . MASTITIS (BOVINE) Diagnosis Clinical mastitis History. Regional veterinary or dairy livestock officers may provide assistance in these investigations. Cost of cell counts depends upon whether dairy farmer is a herd recording member or not. Bacteriology and antibiotic sensitivity testing is performed by the Regional Veterinary Laboratory. ii. Serratia spp and Klebsiella spp can cause persistent mastitis symptoms.i. History required All mastitis submissions should be accompanied by full history of the mastitis case. If a herd problem of clinical mastitis is occurring. Nocardia spp and Arcanobacterium (Actinomyces) pyogenes. increased milk electrical conductivity. previous antibiotic treatment (both for previous clinicals and dry cow treatment). Recovery of milk quality in quarters affected by these pathogens may not occur. VETERINARY LABORATORY MANUAL Mixed quarter samples are not recommended as excessive dilution of the pathogen can occur. with roots attached. Heterozygote detection i. Antibiotic sensitivity necessary for recurrent cases. Of the current S4 antibiotics registered for clinical and subclinical mastitis. If antibiotic treatment has been given. If other antibiotic products are used. antibiotic sensitivity will be necessary. stage of lactation. environmental factors such as contaminated water supply or muddy conditions. cloxacillin products are effective for all staphylococcal and streptococcal infections and neomycinbased products effective for staphylococcal and enviromental mastitis pathogens such as coliforms. Mixed quarter milk cell counts (individual cow cell counts) in excess of 250. Rapid Mastitis Test. age. Recurrent cases of mastitis are usually due to chronic staphylococcal infection. Recovery of the former two pathogens from mastitis cases should indicate cull for slaughter of the affected cow especially if Nocardia sp are involved as these pathogens pose a potential human health risk.

MENANGLE VIRUS INFECTION Menangle virus infection was first recognised when it caused a single outbreak of reproductive disease in pigs in a large piggery near Sydney in 1997. Mannheimia (Pasteurella) haemolytica. Clostridium perfringens is associated with gangrenous mastitis (also S aureus and E coli). Menangle virus infection is notifiable. and is closely related to Tioman virus. Cell counts and NAGase are unreliable in herds with caprine arthritis encephalitis (CAE)virus infection. chronic or subclinical. isolated in 1999 from fruit bats in Malaysia. Fees for tests undertaken to confirm or exclude a diagnosis of Menangle virus infection are paid by NSW Department of Primary Industries.MASTITIS (CAPRINE) Diagnosis Bacteriology is the most effective diagnostic method. Care must be taken with treatment. Clinical signs include a severe decline in farrowing rate. Flying foxes (fruit bats) near the piggery had antibodies to the Menangle virus. A control program. a high incidence of delayed returns to service and a marked increase in the number of mummified and stillborn piglets. subsequently eradicated the virus from the piggery. Streptococcus spp. Micrococcus spp and E coli may also be incriminated in subclinical disease. Stillborn piglets had a severe nonsuppurative encephalomyelitis. It is believed that flying foxes are the natural host of the virus. the virus was not highly contagious and thus probably spread MASTITIS (OVINE) Ovine mastitis may be acute/severe. some with severe deformities. and Streptococcus spp. Diagnosis The cause or causes of the syndrome are not clearly understood. Less commonly Corynebacterium pseudotuberculosis. Other tests including Somatic cell count (high variability makes interpretation difficult. Clinical mastitis can be caused by coagulase negative staphylococci. fever and vaginal discharges in the sow. Staphylococcus aureus. Intramammary antibiotics will have extended witholding periods in goats compared to cows. The most frequently isolated bacteria are Staphylococcus aureus. toxaemia. Parenteral antibiotics may be used. Pseudomonas spp. successfully managed by NSW Agriculture. also epithelial cell shedding as a normal phenomenon increases SCC thresholds for identifying mastitic milk). Actinobacillus seminis. Coagulase-negative Staphylococci. Specimens required Samples of vaginal discharges are of limited value because of the wide range of organisms normally present in the vagina of the sow. The blue dye in these antibiotics can be excreted in the milk of treated goats for extended periods of time. Histophilus ovis. but which has not yet been associated with disease in any animal species. History and specimens required As for bovine samples. gangrenous. Rapid Mastitis Test can be used. and that it was a relatively rare event that led to the virus 'jumping' into pigs. Interpretation of results Coagulase-negative staphylococci (Staphylococcus epidermidis) are the most common cause of subclinical mastitis in dairy goats. A large serological survey failed to find any evidence of infection in other Australian pigs. Menangle virus is a member of the paramyxovirus family of viruses. Bacillus spp. MASTITIS METRITIS AGALACTIA SYNDROME IN SOWS Occurs from 12 to 48 hours after parturition with a rapid fall off in milk production. VETERINARY LABORATORY MANUAL 73 . Management factors are implicated. and NAGase can be applied on a herd basis. and enterobacteria such as Escherichia coli. Serratia spp and Klebsiella spp. Mycoplasma spp can also cause ovine mastitis. constipation. No further outbreaks of the disease have occurred to date. Arcanobacterium pyogenes. Although many pigs were infected. Diagnosis Bacteriology. At present. Rapid mastitis test. there is little that can be offered in the laboratory in relation to this condition. History and specimens required As for bovine samples. Proteus spp and Klebsiella spp are involved.

Specimens required i. histopathology. in buffered formalin for histopathology. Kidney and liver submitted chilled or frozen for selenium estimation. clinical virology. Chilled stillborn piglets for necropsy. iii. ii.e. virology and histopathology. submit heparinised blood from 3 5 peer animals rather than tissues of dead cases Diagnosis History. ii. or indirectly by analysis of blood glutathione peroxidase concentration. and distension of the pericardium by a serofibrinous transudate. 10 ml of heparinised blood. Liver necrosis/ rupture/ haemorrhage is a feature in many cases. lung. iii. as in most enzyme tests. rather than by respiratory aerosols. from at least 5 animals in the group. but these are not the sole cause. submitted chilled and protected from light immediately following collection.by excretion in faeces and urine. 10 ml of heparinised blood submitted chilled (not frozen) for glutathione peroxidase (GSHPx) concentrations. hypomagnesaemia. signs. submitted chilled for virology. The correlation between glutathione peroxidase (GSHPx) and selenium concentrations is very high in nondeficient animals. in buffered formalin for histopathology. Diagnosis History. (See Collection of specimens for biochemistry and Vitamin deficiency). ‘muscular dystrophy’’ This disease complex usually occurs as defined clinical and pathological syndromes which bear a several names. 20 g liver. requires heparinized blood (EDTA is unsuitable). this high mathematical correlation is not present. Fresh heart. Whenever possible. Many cases are associated with vitamin E and/or selenium deficiency. abortion. Sections of affected and unaffected skeletal muscle and cardiac muscle. Selenium status can be measured directly by analysis of blood or tissue selenium concentrations. NB. biochemistry: guide to biochemical parameters in sheep and cattle MUCOSAL DISEASE See Pestivirus. Specimens required i. iv. Specimens required i. liver. white muscle disease. Diagnosis History and characteristic necropsy findings of petechial and ecchymotic haemorrhages in the heart. arthrogryposis and hydranencephaly' MULBERRY HEART DISEASE A cause of sporadic mortalities in rapidly growing pigs. response to therapy. spleen and whole brain in buffered formalin for histopathology. ii. lung. histopathology. brain and body fluids collected separately and aseptically. METABOLIC DISEASES See 'hypocalcaemia. i. kidney. submitted chilled (not frozen) for biochemistry from 5 to 10 animals in the same pen. There was also a severe influenza-like illness in two of the workers at the piggery. nutritional myopathy. VETERINARY LABORATORY MANUAL 74 . free of cells and haemolysis. post mortem examination. They are thought to have a nutritional pathogenesis involving a dietary deficiency of selenium and/or vitamin E. MUSCULAR DEGENERATION. More cases tend to occur under conditions of heat stress. biochemistry. but of little diagnostic consequence. NUTRITIONAL Syn: Selenium/vitamin E deficiency. liver and brain. in deficient animals. Sections of heart. At least 2 ml of serum. submitted frozen for serum enzymology. The two workers recovered from the illness. In deficient animals. The latter. iii. clinical signs. spleen. both GSHPx and Se concentrations are low but there is not necessarily a close mathematical correlation between values. characterised by pronounced myocardial haemorrhage. Heart. hyophosphataemia.

50 days). (DD: Glasser's disease. It has no pneumonic potential and can rapidly overgrow M. Glasser's disease polyserositis or respiratory disease. autumn and spring troughs of Se availability). Swab materials can VETERINARY LABORATORY MANUAL (DD: Erysipelas. Avoid submission of swabs for mycoplasmal culture. This organism is readily cultured. leg weakness/osteochondrosis) Cattle. hyopneumoniae herd infection by ELISA serology. hyopneumoniae cultures. sheep. Diagnosis Pigs Culture of some pathogenic mycoplasma species can be undertaken using media specific for pig mycoplasmas. It can cause a serofibrinous polyserositis.Interpretation of blood glutathione peroxidase (GSHPx) concentrations The following values should be taken only as a guide: GSHPx U/g Hb Sheep <10 <25 >50 be toxic to mycoplasmas if left in contact for extended periods. adult sheep approx. Cattle < 10 < 25 > 100 MYCOPLASMOSIS See also Porcine enzootic pneumonia Mycoplasma infections cause a wide range of clinical conditions. It can also cause outbreaks of bovine mastitis and abortion. • Age of animals preruminant animals more prone to deficiency than ruminants. and can outgrow pathogenic mycoplasmas in broth culture procedures. and caprine and ovine arthritis and mastitis. clover).g. Older sheep normally have lower levels. calves and lambs approx. hyosynoviae arthritis causes synovitis and Interpretation Disease Production response Normal Note that current blood concentration of GSHPx reflect the selenium intake of the animal at the time the present circulating erythrocytes were formed. • Fertilizer history (competitive interaction from sulphur results in reduced selenium in pastures on superphosphate fertilized soils). cattle. Identification of M.g. other Streptococci causing purulent arthritis with percutaneous infections/abrasions. An ‘average’ time would be the half life of the erythrocyte for the species and age of animals to be examined (e. and can occasionally affect young adults. more sporadic disease than Glasser's disease. adult cattle approx. • Pasture type (legumes and other leafy plants lower in Se than grasses). 75 . Pasteurella spp synovitis. Erysipelas). 60 days. Streptococcus suis purulent arthritis or polyserositis. • Soil type (acid soils < pH 5. and is a common co-inhabitant of pneumonic lungs. Streptococcus suis. hyorhinis polyserositis of young pigs. or fibrinous arthritis in suckers or weaners less than 10 weeks old. particularly among pigs. Mycoplasma spp bovine Group 7 is a frequent cause of polyarthritis in calves. hyorhinis is a common inhabitant of the URT and ears (Eustachian tube). special media is required for growth of mycoplasmas.5 have 50% reduction in Se availability to plants). NB: M. goats. In general. Identification of M. hyopneumoniae in lung tissues or nasal swabs by PCR. Diagnosis should be considered on a property basis. eye lesions. For diagnosis of selenium deficiency in sheep. Some mycoplasmas are slow growing. reproductive tract infections. 80 days. It produces a milder. taking up to 3 weeks to appear in primary culture or in subcultures. M. The interpretation of selenium status and prognosis is further complicated by factors which include: • Season (winter and summer peaks. it is recommended that weaners be sampled after access to good feed (e. Certain mycoplasmas are part of normal mucosal flora. M. sheep and goats Culture of affected tissue/s. Other Mycoplasma spp are associated with bovine mastitis. and poultry. and is occasionally cultivable on blood agar cultures without special mycoplasmal media.

Mycotoxin analysis in feed is a specialised area of chemistry not offered by NSW Department of Primary Industries. Often. Demonstration of a toxigenic species of fungus in the feed is not enough. Portion of lesion submitted chilled for bacteriology. such specimens should be stored and transported between 18oC and 37oC. Serum (or heparin plasma) samples for M. Cattle. tremorgen intoxication. hyopneumoniae PCR (1 cm³). the fungi are ubiquitous and only some strains are toxigenic. ergotism. hyopneumoniae PCR. in buffered formalin for histopathology. Mycotoxicosis can only be confirmed by demonstrating biologically effective VETERINARY LABORATORY MANUAL NECROBACILLOSIS Syn: Calf diphtheria. sheep. iii. Section of affected nasal mucosa. joint membrane. Specimens for differential diagnosis. Poultry i. Nasal swab material collected into sterile PBS for M. ii. multiple large creamy/caseous necrotic liver and lung lesions (and occasionally diaphragmatic lesions) are associated with entry of Fusobacterium necrophorum via primary lesions in the rumen and/or abomasum. Specimens required i. NASAL GRANULOMA OF CATTLE Diagnosis Clinical signs. necrotic stomatitis of calves. In lambs.g. Section of affected nasal mucosa. v. Serum for Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) antibody detection by rapid plate test. Specialised services are available at some other laboratories. paspalum staggers. Specimens required i. goats i. histopathology and demonstration of toxic concentrations of mycotoxin in feed. necropsy findings and demonstration of Fusobacterium necrophorum in lesions. because of the expense of mycotoxin analysis. *Tissues for fungal culture should not be refrigerated or frozen (temperatures of less o than 15 C can be detrimental to fungal survival). Fresh chilled joint fluid or membrane for M. Fixed lung for histopathology. joint fluid. hyosynoviae culture. Fresh chilled tissues or joints affected with polyserositis/arthritis for M. 76 . facial eczema. the diagnosis is made by elimination of other causes. the quantity of mycotoxin ingested and the period of exposure. ocular fluid.Specimens required Pigs i. NB Culture for avian mycoplasmas is not routinely undertaken. hyorhinis culture (suckers. Specimens required i. lupinosis and mouldy corn poisoning. ii. ii. in buffered formalin for histopathology to demonstrate toxic effects. (see Nervous disorders). Fusarium spp toxicosis. vi. clinical signs. Fresh chilled or frozen lung for M. Sections of liver and kidney. weaners). milk for mycoplasmal culture. 'necrobacillosis of lambs' Diagnosis Clinical signs. iii. Our Laboratories will outsource this testing on request.. Fusarium spp. 500g of feed for mycotoxin analysis. Depending on the pathogen. ii. concentrations of toxins in the feed the animals ingested. ii. Smears taken from the depth of the lesions. histopathology. hyopneumoniae ELISA. fresh tissues. Diagnosis History. iv. MYCOTIC DERMATITIS See Dermatophilosis MYCOTOXICOSIS The effects of mycotoxins vary according to species and age of the animal. e. particularly where nervous system disease is seen clinically. submitted fresh for bacteriology and mycology*. Sections of affected organs in buffered formalin for histopathology. Mycotoxicoses include aflatoxicosis.

• Neurological disease in the dog (rarely) Diagnosis Cattle History. or are infected transplacentally. Spinal cord and brain and other tissue formalin-fixed for histopathology. NEOSPOROSIS Neospora caninum is a coccidian parasite related to Toxoplasma gondii. The sections should be taken both from the centre of the lesion and also from around the margin. Laboratory diagnosis of the cause or causes will require a range of specimens to be submitted. especially in those cases where the field veterinarian is seeking a prognosis. Diagnosis History. Neospora abortion is reported in deer (rarely). Sections of affected organs. Diagnosis (abortion). sex. submitted in buffered formalin for histopathology. Cattle (an intermediate host species) are infected by ingesting feed contaminated by sporulated N caninum oocysts shed by dogs or by infected cattle tissues (eg infected foetal membranes). particularly: • serous foetal body fluid (pericardial. thoracic or peritoneal) or heart blood for serology • brain and myocardium (including from autolyzed foetuses) formalinfixed for histopathology Cows i. cause: • Abortion in cattle (world-wide. In view of the widespread prevalence of subclinical neosporosis in cattle in coastal NSW areas. including transmissible spongiform encephalopathy (TSE). in buffered formalin for histopathology. with sexual stages in the gut resulting in oocysts shed in faeces. viral. NEOPLASMS In cases of biopsies from the live animal. The history should include species. age. and specific disease syndromes. breed and full description of the tumour giving its location and the rate of development of the tumour. Dogs i. Specimens required Cattle Aborted foetuses i.au/a ahc/programs/adsp/tsefap/tsefap_home. parasitic and protozoon infections.animalhealthaustralia. For adult sheep and cattle with progressive neurological disease. seropositivity in an aborted cow does not confirm Neospora abortion. Nervous signs can arise from a wide range of causes. Specimens as required for diagnosis of abortion in cattle (see Abortion in cattle). NERVOUS DISORDERS See also Ataxia'. Specimens required i. VETERINARY LABORATORY MANUAL 77 . Distinctive histological changes are seen in brain (necrogranulomatous encephalitis) and myocardium (non-suppurative myocarditis) of aborted foetuses. A group of aborted and non-aborted animal should be tested. encephalomyelitis. 10 ml of clotted blood in vacuum tube for serology. ii. clinical signs histopathology. accompanied by a completed NTSESP Clinical History and Postmortem Report form. serology. at the junction of normal and abnormal tissue. Dog Clinical signs (neurological histopathology.iii. please submit specimens as required by the National TSE Surveillance Program (http://www. The asexual proliferative stages of Neospora caninum (including tissue cysts). Clotted blood in vacuum tube for serology. The dog is the definitive host. together with a detailed history and clinical examination. disease). chemical and plant poisonings and metabolic and genetic conditions. serology. including coastal NSW) and congenital neonatal neurological disease in cattle (rarely). a detailed description is necessary. Sections of affected organs.cf m). gross pathology and histopathology. clinical signs.com. including bacterial.

NITRATE-NITRITE POISONING Diagnosis History. Range of fresh and fixed tissues for differential diagnosis. Portion of brain. Methacrifos. submitted chilled or frozen for enzymology and biochemistry. ii. Methyl Parathion. Specimens required i. Gamma Chlordane. iv. or. submitted chilled for serology. chemical confirmation of nitrate or nitrite in the animal. in buffered formalin for histopathology. OPHTHALMIA Syn: Pink-Eye Diagnosis Clinical signs. Branhamella spp or Chlamydia in sheep). The history should include details of the specific insecticides to which the animals may have had access (see also 'Specimens for Toxicology'). Specimens required Either. Thick. identification of causal organisms (Moraxella bovis or IBR in cattle. Brain and sections of jejunum and ileum in buffered formalin for histopathology. Organophosphates Bromophos Ethyl. ORGANOCHLORINE AND ORGANOPHOSPHATE POISONING There may be some delay (approximately 2 weeks) in testing samples for organochlorines and organophosphates. spinal cord and cerebrospinal fluid. Arrangements can be made with the laboratory for the supply of blood agar plates in special circumstances where the isolation of Moraxella bovis is desirable. NB. Diazinon. submitted chilled for bacteriology. Moraxella bovis can be recovered consistently from the conjunctival sac only when blood agar plates are inoculated immediately swabs are taken. 1 ml of serum submitted chilled for toxicology. and others. together with paired sera. Fenitrothion. Parathion. Brain and spinal cord. Appropriate specimens should also be taken to eliminate other causes of sudden death. Segment of ileum or swab in transport medium for bacteriology. Specimens required OEDEMA DISEASE OF PIGS Diagnosis Clinical signs. Ethion. Dieldrin. At least 3 ml of serum. necropsy findings and the recovery of enteropathogenic Escherichia coli. Sulprofos. Specimens required i. clinical signs. Other specimens as required for specific diseases. Specimens required i. submitted chilled for virology. Diagnosis Based on clinical findings. There are no pathological changes observed at necropsy. Pirimiphos Methyl. Serum sample from the affected animal. i. clinical signs. Fenthion. DDE. air dried blood smears for toxicology. Endrin. DDD. Chlorpyrifos Methyl. iv. histopathology and biochemistry. v. iii. Chlorpyrifos. serum or blood is best performed in the field. Swabs of conjunctival sac in phosphate buffered gelatin saline (PBGS). iii. response to treatment and toxicology. Alpha BHC and Beta BHC. HCB. Heptachlor Epoxide.History. Alpha Chlordane. ii. Profenophos. DDT. collected aseptically and submitted chilled for bacteriology and virology. Currently analytical methods for pesticides in ingesta or tissues are available for the following chemicals: Organochlorines Aldrin. Malathion. Testing is undertaken at NSW Department of Primary Industries' laboratory at Wollongbar for a range of OC and OP compounds. Smears and swabs of conjunctival surface. Testing of plant material. free of cells and haemolysis. Moribund or freshly dead animal for necropsy and bacteriological examination. Dichlorvos. particularly methaemoglobinaemia. ii. Chlorfenvinphos. ii. Heptachlor. VETERINARY LABORATORY MANUAL 78 . Lindane.

joint pigs. Dead animal i. Specimens required i. PAPULAR STOMATITIS OF CALVES Diagnosis Clinical signs.Acute exposures i. At least 2 ml of serum. Specimens required Live animal i. Specimens required Affected limbs. porous and soft. and often the seat of healing fractures. histopathology. should be submitted in buffered formalin for differential diagnosis on histopathology. free of haemolysis and cells. ii. with both ends tied off (also tied at the abomasoduodenal and ileocaecal junctions). (see also Hypocalcaemia). Chronic exposures i. liver and brain submitted frozen for toxicology. VETERINARY LABORATORY MANUAL PARAKERATOSIS OF SWINE Diagnosis Clinical signs mange and histopathology. At least 30 g of fresh faeces as above. brain and any other organs with gross lesions. submitted chilled for electron microscopy. clinical findings. submitted chilled for egg count and larval culture. submitted in buffered formalin for histopathology. differentiation sarcoptic exudative epidermitis. gross pathology and histopathology. kidney. brain and any other organs with gross lesions. bone fractures. submitted chilled for calcium estimation. ii. gross pathology. clinical signs and gross pathology. histopathology. Most cases are best diagnosed on clinical signs and gross pathology. faecal egg count and larval culture. hypocalcaemia. In cattle. serum or plasma pepsinogen levels (see Pepsinogen estimations from serum or plasma'). Digestive tract lesions fixed in buffered formalin for histopathology. Affected bones are light. if required for differential diagnosis. Sections of affected bone. 79 . with unopened joints. Scabs or necrotic debris in a sealed container. submitted in buffered formalin for histopathology. In cattle. Specimens required i. serum or plasma pepsinogen may be useful. At least 20 g of stomach contents. At least 30 g of fresh faeces in a screwtop jar filled to capacity. electron microscopy. submitted chilled for bacteriology. age of affected animals. OSTEOMALACIA. Sections of liver and kidney. perverted appetite and tendency to bone chewing. ii. should be submitted in buffered formalin for differential diagnosis on histopathology. OSTEOCHONDROSIS IN PIGS A condition characterised by abnormalities in rapidly growing usually seen in baconers. OVINE BRUCELLOSIS See brucellosis ovine' OXALATE POISONING Diagnosis History of access to plants containing oxalate. following laboratory consultation Specimens required i. The abomasum and abomasal contents whole. Diagnosis History. kidney. submitted chilled or frozen for total worm count and abomasal digest for recovery of histotrophic larvae. Sections of liver. Diagnosis Clinical signs. lung. Histological examination is performed only in certain cases. clinical examination. ii. Sections of liver. OSTERTAGIOSIS See also Parasites internal Diagnosis Grazing history. At least 20 g of fat submitted frozen for toxicology ii. lung. ii. total worm counts. OSTEOPOROSIS Disease associated with transient and shifting lameness. particularly in growing animals.

faecal egg count. PARASITES (EXTERNAL) See also Itchmite. ii. unopened and tied off at each end.nsw. Milk ELISA testing for liver fluke is currently (2004) under validation. WormTest is a convenient system for collecting and submitting faecal samples to the laboratory. iii. NB Faecal egg counts are of no value in diagnosing clinical paramphistomiasis. Insect larvae . ii. submitted in buffered formalin for histopathology. pasture availability. can be collected from "struck" wool by placing the wool on a sheet of paper in sunlight. etc. paddock movement. ii.au/reader/sheepinternal http://www. Further information on internal parasites of sheep or cattle is available on: http://www. The abomasum and small intestine. Skin scrapings. Specimens required Live animal i. submit preserved in alcohol/ glycerine (9 parts of 80 per cent alcohol to 1 part glycerine) in a small leak proof container. The presence of paramphistome eggs indicates non-pathogenic adult stomach flukes are present. In cases where parasites cannot be demonstrated in skin scrapings in the field. a skin biopsy sample from an affected area. 10 mL clotted blood tube or 2 mL serum for zinc analysis. Specimens required for parasite identification i. PARASITES (INTERNAL) See also Ostertagiosis. submitted in a screw top jar filled to capacity for egg count and larval culture. In flock or herd investigations. lice. paramphistomiasis' Faecal egg counts and differential larval counts are a guide to the size and type of worm burden.Specimens required i.nsw. larval cultures and total worm count. At least 30 g of faeces collected from the rectum. Larvae VETERINARY LABORATORY MANUAL Dead animal 80 . Specimens required Dead animal i. etc.gov.scrape recent lesions. e. tongue and affected skin. stage of pregnancy and effects of lactation.agric. ticks. submitted in buffered formalin for histopathology. serum sample chilled for ELISA serology. samples should be collected from 10 to 20 animals showing evidence of parasitism. Portions of oesophagus. stocking rates. liver fluke infection. consign about fifty in a ventilated container with about 3 cm damp vermiculite in the bottom.where live larvae are required for insecticide tests. Mange mites . Flies. spiders. Adult paramphistomes may confer some immunity. mange' Diagnosis Based on clinical findings and demonstration and identification of parasite on skin scraping or skin biopsy. submitted chilled or frozen for parasitology. PARAMPHISTOMIASIS See also Parasites internal' Diagnosis Grazing history and associated seasonal conditions. The history should provide details of all recent anthelmintic treatments and flock management. For F hepatica in cattle. ii. demonstration of immature paramphistomes in the duodenum or adult flukes in the rumen and reticulum. fleas. Transfer scrapings to a small wide mouthed bottle and submit unpreserved.au/reader/cattlehe alth Diagnosis Clinical signs. as well as the parasite involved and whether the parasites are sexually mature. submitted unpreserved for parasitological examination.g. swampy areas. The faecal egg count depends on a number of factors including faecal consistency and bulk. host resistance.gov. check with your veterinary laboratory for availability.agric. Clinical findings. Specimens required for diagnosis i. using a scalpel moistened with liquid paraffin.

humidity) and where the biotic potential is high. Any statement of 'significant figures' for ova counts will be only a rough guide. The number of helminth ova passed per gram of faeces depends on such factors as faecal consistency and bulk. Sheep An egg count of 500 eggs per gram (epg) is generally considered high enough to require treatment in order to limit pasture contamination and subclinical disease. Alternatively i. state of nutrition and clinical history of the infected animals. 500 1. Hillard JJ (1966) p7.000 Oesophagostomum columbianum 1. Low and medium egg counts will be more significant where the stocking rate is high. e.Preferably: i. Skerman KD. 500 Trichostrongylus spp. the count may be increased.g. The whole gastrointestinal tract (double bagged in strong clear plastic) submitted unopened. Inappetence may cause the count to multiply 30 to 40 times. See AgNote: WormTest for livestock and guide to egg counts. when weather conditions are conducive to epidemics (warmth. Cattle The clinical history and knowledge of the seasonal pattern of worm parasites in different areas of the State will assist in interpretation of faecal egg counts. Fasciola hepatica and paramphistomes. submitted in a 5% formalin solution for parasitological examination. FAECAL EGG COUNTS (INTERPRETATION) Faecal worm egg counts and differential larval counts are a guide to the parasite burden.000 Haemonchus contortus Ostertagia spp. the egg count gives little indication of the level of the parasite burden. Haemonchus contortus. In cattle greater than 18 months of age. If sheep are starved for 24 hours.000 Chabertia ovina >100 Fasciola hepatica Paramphistomes >500 *Compiled from various reference sources: Cole VG (1986) Appendix 1. Chabertia. submitted chilled or frozen for examination for immature parasites. Hutchinson GW (2003) Table 4. effects of lactation and whether the worm burden consists of sexually mature parasites. stage of pregnancy.000 500 500 >200 300 500 100 2. 1. Interpretation must be considered in relation to the specific parasite responsible (as indicated by larval cultures) and the age. ii. This is of particular significance with Nematodirus. host resistance. origin. Aliquot samples of 1 L from the stomach and small intestines. Guide to faecal egg counts in sheep (indicating pathogenic burdens) Species Eggs per gram of faeces Young Sheep Older Sheep 2. Diarrhoea depresses the egg count. VETERINARY LABORATORY MANUAL 81 . The egg laying capacity of Ostertagia spp and Nematodirus spp is poor and severe clinical signs may be seen before appreciable numbers of eggs are present in the faeces. Ostertagia. The abomasum and intestine. rain. Faecal egg counts are generally lower from cattle than from sheep. Love SCJ. but tied off. Infections with one parasite only are rarely seen and the additive effects of mixed infections will require assessment. The following numbers of epg may indicate clinical disease due to that parasite. pp 329-333. Dictyocaulus filaria is often associated with mixed gastrointestinal infections on the tablelands and slopes.000 Nematodirus spp. The pathogenicity of immature stages not indicated by egg count should always be considered. either chilled or frozen for parasitological examination. pp 233-239.

e.000 worms is considered a light burden. there is little relation between egg count and fluke burden. In young sheep 1. 9. Ostertagia spp.00010. ill thrift and sometimes mortality. Opinions vary on the significance of counts for various worm species. Colic induced by verminous arteritis from migrating Strongylus vulgaris larvae (without eggs in faeces). 20.000 2. Smeal MG (1995) pp 358. In young sheep 500 worms (light). In the absence of worm egg counts.000 worms is a heavy infection in young sheep. 82 VETERINARY LABORATORY MANUAL . Skerman KD. A similar number of worms in adult crossbred sheep can be responsible for ill thrift and scouring but probably 30. Usually there is a concurrent infection with Trichostrongylus spp.00010.000 worms are needed to cause mortalities. the importance of immature stages of Nematodirus spp. Heavy infections (>5. Pigs Any positive count of Ascaris suum ova in pigs up to 5 months of age is significant.500 Haemonchus placei Cooperia spp (in 3-4 month old calves) 1.Guide to faecal egg counts in cattle (indicating possible pathogenic burdens in 6-18 month old cattle)* Species Ostertagia spp. and other intestinal worms. Horses 600 epg or more may represent a pathogenic cyathostomin (small strongyle) burden.g.000 (heavy) and in adults 1. Heavy infections of adults.000-30.000 worms may cause mortality. eggs generally appear after the age of 9 to 10 weeks. but heavier infections of 5. but depends on clinical history and several interacting factors such as age.000 5.000 worms may be an important cause of diarrhoea. Small intestine Trichostrongylus spp.000 worms can be considered a heavy infection. 5000 paramphistomes may be associated with ill health. 3.000 worms in adult animals may cause mortality. while in older weaners.000 (light).500 (<300 possibly significant) 500-1. The immature forms occur in the small intestine.000 in acute disease) Oesophagostomum radiatum. In infected pigs.000 40. Trichostrongylus axei. Abomasum Haemonchus contortus. 3. Nematodirus spp. Hillard JJ (1966) p 8. Sheep Rumen and Reticulum Paramphistomes. 5. 5.000 worms) may cause unthriftiness and gastritis. pp240-245. Young pigs with heavy immature worm burdens may not be passing any eggs in the faeces. Paramphistomes >500 (heavy adult infection) *Compiled from various reference sources: Cole VG (1986) Appendix 2. Hutchinson GW (2003) Table 5.000-5. sex and nutritional status of the host. Eggs per gram of faeces 300->1. In young poorly grown weaners.000 (10. However. is not as common as in previous times.000 Trichostrongylus axei (medium infections) 700->1.000 (heavy). Love SCJ. TOTAL WORM COUNTS (INTERPRETATION) Interpretation of total and differential worm counts is not absolute. >500 Bunostomum spp 500-800 (heavy infections) Fasciola hepatica Any egg count is significant Heavy infections may be indicated by >25 epg. pp 334-338 . Usually observed as a concurrent infection with Trichostrongylus spp. should be considered in the diagnosis of clinical parasitism.

foetid scour. and sometimes diarrhoea. e. Heavy VETERINARY LABORATORY MANUAL 83 . Nematodirus spp. but in lower numbers. Moniezia importance. Rarely found in large numbers. This hookworm causes anaemia. In both young and adult sheep. The number of immature stages in the histotrophic form in the abomasal wall should be considered when planning anthelmintic treatments against outbreaks of ostertagiosis. Chabertia ovina. or very marked nodule formation in previously exposed older sheep due to Oesophagostomum columbianum. There may be a heavy infection with larval forms. Greater than 50-100 worms may be considered a serious burden.g. 50. In 6-month-old calves. Cooperia spp. 5. weight loss.000-50. haemorrhagic enteritis and dark. when adults may be rare and egg count consequently low. Heavy burdens of more than 200.000-90.000 worms).000 paramphistomes. Following a single anthelmintic treatment which removes adult worms. The most common small intestinal worm. Infections with > 5. Trichostrongylus axei. Liver Fasciola hepatica. (80% of immatures are in duodenum). Usually found in association with Ostertagia ostertagi. Inflammatory lesions in the caecum resulting from large numbers of parasites probably cause scouring and ill thrift. Mortalities from the acute disease may be associated with 700 or more immature liver fluke and from the chronic disease with 50 or more adult fluke.000 adult worms may cause diarrhoea. Always occurs in mixed infections with Ostertagia ostertagi. anaemia and harsh coat. Both immature and adult worms can cause ill effects. Lungs Lung worms (Dictyocaulus filaria. 5. submandibular oedema. Moniezia spp.000 adult worms may be pathogenic causing anaemia. Less important in cattle than H contortus is in sheep. spp. Rarely found in large numbers. Common in calves but relatively unimportant.Strongyloides spp. The immature forms occur in the small intestine. In young sheep 100 worms is a serious infection and in adult animals 100-200 worms may be significant.000 immature stomach fluke cause clinical disease. 100-200 worms is a heavy infection. may be associated with ill health. Ostertagia ostertagi. including bulls. Ostertagiosis should be considered in the differential diagnosis of ill thrift and loss of condition in adult cattle. Low numbers (2.000 immature worms could easily cause a second serious wave of adult infection (Type II ostertagiosis).000 worms may occur in dairy calves. Not considered of pathogenic importance under field conditions except in young lambs when infection is heavy (10. Trichuris ovis.000-15. Heavy infections have been observed in sheep during prolonged drought periods. Serious chronic effects are caused by severe nodule formation. Heavy infections in young lambs may be of importance. Trichostrongylus spp. Heavy infections may be seen in dairy calves (10. Bunostomum phlebotomum. May be a problem under penned conditions. Cattle Rumen and Reticulum Paramphistomes. Abomasum Haemonchus placei.000). In young calves 500 worms may be significant. Small intestine Cooperia spp. Questionable infections of adults.000-3.000) may be found in young cattle in concurrent Large intestine Oesophagostomum spp. Muellerius capillaris) are relatively rare in sheep. An important pathogenic parasite of both young and adult cattle as 30. but clinical signs are due primarily to ostertagiosis. Paramphistomes. the ensuing development of up to. Oesophagostomum columbianum is usually restricted to northern and western district pastoral areas of NSW.

Proceedings 350. Sydney. Heavy infections may affect young suckers. Macracanthorhynchus spp. Usually found only in low numbers and is of little pathogenic importance. UNDP. A Handbook for Studies of Helminth Parasites of Ruminants. Heavy infections can cause typhlitis in young pigs. Paramphistomes. Infections with >10. Lungs Dictyocaulus viviparus.infections with trichostrongyles. but occasionally causes stillbirths if infection takes place before 70 days gestation. with returns to service. Parasites of Cattle. Post Graduate Foundation in Veterinary Science. Large intestine Oesophagostomum radiatum. Whole mummified foetuses and stillborn foetuses submitted chilled for bacteriology. other intestinal trend to free-range pigs continues. AGPS. stillborn and weak piglets. The prepatent stages of this worm in the intestinal wall may also cause ill effects. Stephanurus dentatus (kidney worm) very rare parasite in modern pig husbandry.000 immature stomach fluke cause clinical disease.anaemia. ii. but still found in feral pigs in some locations. but seldom seen in beef cattle. and a watery. Love SCJ. Near East Animal Health Institute. Infection from 35 to 55 days of gestation results in foetal death and mummification. Very pathogenic. Liver Fasciola hepatica. Serology on individual samples from sows is of little value in diagnosing parvovirus infection. An important cause of liver condemnations at abattoirs. Smeal MG (1995). FAO. Australian Agricultural Health and Quarantine Service. In Gross Pathology of Ruminants. Common in dairy calves in cooler areas. Not common. haemorrhagic enteritis. May be re-introduced if VETERINARY LABORATORY MANUAL PARVOVIRUS INFECTION IN PIGS Infection before 35 days of gestation causes foetal resorption and small litter size. Stephanurus and Strongyloides may be found wandering in the lung. mucoid scour. Prevalent lungworm. Rome. Strongyloides spp. clinical findings. Diagnosis Herd history. Widespread and common cause of scouring in pigs. Pathology and diagnosis of internal parasites in ruminants. Helminth Parasites of Sheep and Cattle. Canberra. Veterinary Review No. Immature forms of Ascaris. Iran Unit. Sydney. Light infection up to 50. A burden of 500-800 worms is a light to medium one. Found in calves 4-12 months old. Abortion is extremely rare in parvovirus infection. Department of Primary Industries. while 1. Animal Health in Australia Volume 8. Pigs Small intestine Ascaris suum. Often seen in conjunction with gastrointestinal parasite burdens.000 worms may produce clinical signs of parasitism . Heavy infection is 200 worms. virology and histopathology. (80% of immature worms are in duodenum). the virus does not cross the placenta. 32. Parvovirus serology should be used as a herd 84 . Skerman KD. References Cole VG (1986).Chapter 16:309-338. liver and kidneys Metastrongylus spp. Hutchinson GW (2003). Trichuris spp. medium infection 50-100 and heavy infection. University of Sydney. Lungs. Post Graduate Foundation in Veterinary Science. Hillard JJ (1966). over 100. Specimens required i. Any number is significant in young pigs. University of Sydney. Between 80 days and term. Infection after 55 days usually has no effect. Heavy and pathogenic burdens of 50100 larvae. histopathology and virology (demonstration of virus antigen or antibody in foetal tissue). Large intestine Oesophagostomum spp. Trichuris spp.

Specimens required i. Pepsinogen values should be interpreted on a herd rather than an individual basis. False negatives may occur. toxigenic strains of P. or EDTA o Activity is stable for several days at 4 C for several days and several months at -20oC. and can be part of a foetal neonatal mortality syndrome. heart blood and any other organ showing lesions. spleen and kidney. spleen. submitted chilled for bacteriology. Serum. Results are compared with a tryrosine standard. ii. It can also cause abortion and mastitis in sheep. Diagnosis Clinical signs and necropsy findings. Abomasal damage results in increased blood pepsinogen concentration. multocida are associated with enzootic atrophic rhinitis (see Atrophic rhinitis of swine). lung. Sections of liver. estimations of serum or plasma pepsinogen concentrations are considered a useful indicator of abomasal damage and of value in the diagnosis of type II ostertagiosis (caused by the synchronised emergence of histotrophic larvae). The assay measures the presence of pepsinogen through the ability of the test serum or plasma to breakdown a protein substrate to peptide fragments. heparinised plasma. Toxigenicity testing of snout isolates from pigs. Pepsinogen is produced in the gastric mucosa as the inactive precursor of pepsin. PERINATAL MORTALITIES IN SHEEP AND GOATS Investigations should be based on a field necropsy with laboratory assistance to confirm or make a specific diagnosis 85 Interpretation of results VETERINARY LABORATORY MANUAL . Histopathology. In pigs. NB. submitted in buffered formalin for histopathology. develops within 5 days of birth) • Mannosidosis (progressive from birth) • Generalised glycogenosis (progressive from birth) • Cardiomyopathy and woolly haircoat syndrome (progressive from birth) Environmental diseases • Mineral deficiencies • Selenium deficiency (WMD. The following conditions should be considered. see specimens required for Abortion. stillbirths occasionally) Infectious diseases • Bacterial Leptospirosis. Recovery of Pasteurella spp.test or to assess the immune status of individuals. and expressed in terms of IU/L of tyrosine. PERINATAL MORTALITIES IN CATTLE Bovine stillbirths and neonatal deaths are often related to causes of abortion. Palyam • Chlamydial PEPSINOGEN ESTIMATIONS FROM SERUM OR PLASMA In adult cattle. and specimens collected according to the disease entity where appropriate: Dystocia Congenital defects Genetic diseases • Maple syrup urine disease (after first feed) • Inherited congenital myoclonus (signs present at birth) • Citrullinaemia (born normal. In cases of abortion. Campylobacter • Viral Pestivirus. * Levels of 5 or greater are considered indicative of damage sufficient to cause production losses.and Mannheimia haemolytica (syn: Pasteurella haemolytica) can cause a septicaemic or pneumonic disease. Specimens required i. Haemolysis has little effect on activity. lung. Level (IU/L) <5 5-10 10-15 > 15 Interpretation No significant abomasal damage Minor damage* Moderate damage Major abomasal damage PASTEURELLOSIS Infection by Pasteurella spp. Sections of liver. Akabane.

including congenital infections • Dystocia • Starvation. Life long carriers of the virus including cases of mucosal disease [MD]) are a consequence of infection in early foetal life (usually before 90 days gestation). nasal swabs in PBGS for virus isolation. perinatal mortality. 10 ml of unclotted blood (heparin blood is preferred). Some stillborn. ii.g. maternal hypocalcaemia. poor growth rates. Immune tolerance to the virus is induced. overfat ewes • Clinical conditions in the ewe likely to result in a prolonged parturition (uterine inertia) and resultant foetal hypoxia or anoxia e. the product of infection at 120-180 days gestation. foot abscess. supported by the laboratory. Preferred specimens are (in order): spleen. poor body condition due to other causes • Diagnosis Flock history. mesenteric lymph 86 VETERINARY LABORATORY MANUAL . In cases of mucosal disease. Detection of antibody in precolostral serum of deformed calves. field investigation. non specific immunosuppression and disease with mortality in calfhood or early adult life. there must be prior arrangement with the laboratory. PESTIVIRUS INFECTION Syn: Mucosal disease. clover disease.g. with resultant inability of lamb to gain sufficient early feed. Post-parturient death iv. Foetal infection which persists into and through post natal life will have resulted from one of two circumstances: • Infection of a susceptible dam. iii. However. Detection of antigen or isolation of virus from blood clot or tissue. may be seropositive. ii. submitted chilled for antigen detection. Notes: • In cases of embryonic mortality or abortion. around 5% of viraemic animals may have antibody to pestivirus. mucosal disease virus infection. Whole lambs or kids submitted chilled for bacteriological and pathological examination. • The dam herself is a persistently viraemic and immuno-tolerant animal. Specimens as required for Abortion in sheep. serum samples from 1015 cows may provide evidence of herd infection and allow more selective investigation to confirm pestivirus involvement. e. lung. Other affected [=viraemic] animals are usually antibody negative. iii. Dead animal i. assuming it is old enough to have lost maternal antibody (> 6 months). • A newborn calf with congenital deformities. congenital malformations. • Delayed colostrum production in ewes suffering nutritional stress in the last month of pregnancy • Predation • Postparturient infections. maternal malnutrition (with foetal anoxia from prolonged parturition). In cases of respiratory disease. pregnancy toxaemia. regarding the number & nature of specimens to be examined. abortion or respiratory disease. bovine viral diarrhoea virus Pestivirus infection is associated with embryonic or foetal death. Conditions to be considered should include: • Abortion. selenium deficiency. A clot (from 10 ml of blood collected into a plain tube) may be used for antigen detection but offers lower sensitivity. Demonstration of an antibody titre in a live animal usually indicates that the animal is not persistently infected. Specimens required Live animal i. probably through contact with a virus carrier. pneumonia and enteritis. perinatal mortality. or. demonstration of virus antigen by ELISA on fresh tissues provides a rapid diagnosis. respiratory disease. including necrobacillosis. apparently-normal calves can also be seropositive.In any detailed investigation. Diagnosis Clinical signs and pathology. Specimens required Abortion and parturient death i. • Nutritional factors. mismothering or exposure • Low birth weight from maternal malnutrition during last 2-3 months of pregnancy. in early (usually 30 100 days) pregnancy. Serum from clotted blood is preferred for antibody detection.

there is no specific diagnostic test available. and Tribulus spp. and kidney may be submitted chilled for virus isolation. ii. PLANT POISONING See Poisoning plant PNEUMONIA Diagnosis Clinical and post mortem examination. Foetal fluids (pericardial. ii. further testing of maternal relatives and enquiry into the history and management of the herd may indicate how or when infection occurred. Pestivirus. Diagnosis Flock history. submitted chilled for microbiology. An EDTA blood haematology. PI3 and RSV). clovers (Trifolium spp. medics (Medicago spp. pleural or peritoneal. PIGLET ANAEMIA Iron deficiency causes a microcytic hypochromic anaemia in sucking piglets. VETERINARY LABORATORY MANUAL Dead or slaughter animals i. Plant material for identification. i.node. Nasal swabs in transport medium for virus isolation. frequently with subsequent necrosis. in order of preference) should also be collected to test for IgG and for pestivirus antibody of the IgG level is elevated. In cases of staggers.). clinical findings. gross pathology. Specimens required Live animal Especially in feedlot or other intensively reared cattle. specimens as required for ataxia. jaundice being an inconstant feature. A number of other plants are incriminated. Bacteriological. See also Protoporphyria (Limousin calves). ii. oedema. virological. sample for PHALARIS POISONING Can cause staggers or sudden death. One or several of these should be submitted. The cadaver submitted chilled for bacteriology and histopathology to investigate a differential diagnosis. spleen. histopathology. Where pestivirus infection and/or mucosal disease has been confirmed. Portion of affected lung. Specimens required i. facial eczema. Options available include removal of carrier families and management to ensure the exposure of breeders to carriers before joining. Plants often causing photosensitization include Hypericum perforatum. Portions of lung. especially from cases of respiratory disease where infections are transient and may not be detected by ELISA. ii. PHOSPHORUS DEFICIENCY See Hypophosphataemia' PHOTOSENSITIZATION This condition is characterised by the occurrence of pruritus. Sections of liver and kidney. Dead animal i. algae and conditions giving rise to liver damage. Other causes of photosensitization are phenothiazine. etc submitted chilled for virology. bowel and parotid salivary gland submitted chilled. Specimens should be submitted to exclude other possible causes of death. pleural fluid and mediastinal lymph nodes. and what steps can be taken to eliminate the problem. iii. histopathological examination. submitted in buffered formalin for histopathology. clinical signs. haematology.). ii. Paired (acute and convalescent) sera for virus serology (for IBR. Diagnosis Mainly on clinical examination. 87 . parasitological. Diagnosis History. trachea. Fresh lung. In cases of sudden death. PINKEYE See Ophthalmia' Specimens required i. District Agronomists may be able to identify suspect plant material. Specimens required Live animal i.

POISONING (CHEMICAL) See also Arsenic. It will only be performed for specific plants to which the animals have had access. Specimens required i. As the analyses tend to be expensive. clients should carefully consider history. specimens as required for Nervous disorders. sodium fluoroacetate. In goats where retrovirus infection is suspected. Diagnosis History. • A limited range of analyses are available on a routine basis. nitrate and oxalate poisoning and pyrrolizidine alkaloidosis Diagnosis History (sudden high morbidity/mortality. clinical signs. Where NSW Department of Primary Industries Laboratories are not capable of conducting the requested analysis. submitted in buffered formalin for histopathology. Sections of liver. IDENTIFICATION OF SUSPECT POISONOUS PLANTS Advice in the first instance should be sought from the local District Agronomist. Most chemical analyses are specific for at best a narrow group of chemicals eg organophosphates.iii. kidney and other organs showing lesions. iii. necropsy findings. showing the leaves in position together with flowers and/or fruits. e. clinical signs. A check can then be made at necropsy to determine if these plants are present in the ingesta. additives) are suspected. In pigs. organochlorine and organophosphate. strychnine and urea poisoning NB Examination is not undertaken on specimens likely to involve criminal prosecutions (e. kidney and any other organs showing lesions. salmonellosis POISONING (PLANT) See also Cyanide. iv. specimens should be submitted as follows Collection of plant specimens All specimens sent should consist of a small branch or portion of the stem. If plant poisoning is suspected. dips. v. If nervous signs are seen. submitted in buffered formalin for histopathology.g. When this is not possible. NB The Specimen Advice Form should request a particular analysis. Specimens required i. the police or other relevant regulatory authority should be consulted before any action is taken (see Conditions for acceptance of specimens). lead. clinical findings and pathological findings so that only the most likely chemical is nominated when completing a request for chemical analysis. submitted in buffered formalin for histopathology iv. Specimens as required to allow a differential diagnosis. drenches. serum sample from affected animal submitted chilled for virology.g. At least 100 g of stomach content. soil for chemical analysis. see porcine pleuropneumonia and porcine enzootic pneumonia (PEP'). and in some cases. Approximately 500 g of suspect material eg feed. and provided that specimens of those plants accompany the specimens of ingesta. Confirmation of chemical poisoning depends upon the demonstration of the poison in body tissues and organs. (see Caprine arthritis encephalitis'). laboratory examination. NB Analysis of this material is usually only requested after poisoning is confirmed and submitters wish to identify the source of the poison. 20 30 cm long. feeds. submitted chilled for toxicology. NB Examination of ingesta is usually difficult and of limited value. Sections of lung. the paddock should be inspected to determine the availability of possible toxic plants. samples will be referred at cost to an outside provider selected on the basis of NATA accreditation and value. evidence that suspected toxic plant has been eaten).g. NB • Irritant poisons will cause a gastroenteritis. ii. in cases of suspected malicious poisoning). In such cases. ii. then material should be submitted as recommended by the manufacturer. • When specific drugs (e. Owing to 88 VETERINARY LABORATORY MANUAL . Sections of liver.

and notes on the main features of the plants and their habitats. POLYARTHRITIS See Arthritis POMPE'S DISEASE between See Generalised glycogenosis Fresh specimens should not be forwarded unless particularly requested. spirochaetal diarrhoea. Labelling of plant specimens Each plant specimens should be numbered and a second set of specimens with corresponding numbers retained. it is difficult. listeriosis. Gross pathology and histopathology of brain. in addition to the adult foliage. in the case of trees. particularly require adequate specimen preparation. The cooperation of inquirers is appreciated in supplying further information or material when rare specimens. while in the tree ferns the scales or hairs at the base of the stalk of the frond are essential for identification. the extent of rough bark when present. Differentiate from tetanus. For small plants and grasses. PORCINE COLITIS Syn: Porcine intestinal spirochaetosis (IS). Nardoo. habit and the habitat are very important. Not more than 12 specimens will be named in one collection. as well details of the habit of growth. precise locality details. the whole of the plant should be sent. Information on the bark type. nervous disease principally of sheep and goats. Specimens will only be returned on special request. focal symmetrical encephalomalacia/ enterotoxaemia. salt poisoning. • Diagnosis Clinical signs (typically progressive higher CNS dysfunction with aimless wandering. blindness. Specimens required i. Specimens forwarded for should be addressed to: identification The Director (Attention: Botanical Inquiry Section) National Herbarium Royal Botanic Gardens SYDNEY 2000 POLIOENCEPHALOMALACIA (PEM) Syn: Cerebrocortical necrosis (CCN) See also: Nervous disorders A sporadically occurring. fruit and juvenile leaves. Storage and despatch of plant specimens Press and dry specimens newspaper sheets. and. Response to 6 10 mg/kg thiamine IV in live affected animals (preferably those not yet recumbent). type of soil. Specimens from other states. or. the rhizome (root like structure) is required. A list of identifying names or information corresponding to the numbers will be reported. submitted in buffered formalin for histopathology. The data and place of collection should be given. height. The disease in ruminants is considered due to either: • production of thiaminases in the rumen following changes in the ruminal flora. non-specific colitis 89 VETERINARY LABORATORY MANUAL . or specimens of special interest. CAE (goats) and copper deficiency induced leukomalacia. less commonly of cattle and occasionally pigs. with characteristic brain lesions in advanced cases of cerebrocortical necrosis. and sometimes impossible to determine specimens from leaves alone. a description of the bark. ingestion of thiaminases in plants (e. recumbency.the tremendous number of plant species. ii. With small ferns and fern allies. rock fern) or in chemicals (eg.g. Specimens as required for diagnosis of other nervous disorders. except under special circumstances. have been identified. convulsions and death). particularly when indigestion occurs after sudden dietary change. amprolium in poultry feed/ manure). flower colour. which may be unfamiliar to local botanists. with the exception of the terrestrial or ground orchids when only the parts above ground should be collected. opisthotonus. Eucalypts can be difficult to identify without samples of the buds. Whole brain. habitat. lead poisoning. head pressing.

It mainly affects weaners 2-6 weeks post weaning but can affect growers. . Trichuriasis Trichuriasis can produce an identical macroscopic mucohaemorrhagic colitis and typhlitis to that of swine dysentery. Short chain fatty acids (SCFA) can cause reversible changes to colonic epithelium. salmonellosis. ad-lib feeding and high density diets may be associated with this condition. scattered mucosal erosions. in which large-intestinal histological. and submucosal congestion. swine dysentery. 90 . producing abnormally high levels of SCFA. and sometimes macroscopic. elimination of infectious agents associated with other syndromes or diseases with similar presenting signs (L intracellularis. persisting a few days. diagnosis of non-specific colitis or niacin deficiency by dietary history and elimination of infectious agents. lesions are seen. associated with a mixed leucocytic. Dietary change. Dietary colitis syndrome (non-specific colitis) Dietary colitis syndrome is a recently described condition of rapidly growing pigs. Fresh colon or caecum or faeces (live animals) for detection of Brachyspira pilosicoli by PCR. Sections of affected intestine (especially ileum and colon). which is usually home-mixed. crypt dilation. The disease is expressed as fluctuating and cyclical in a herd. to be fermented in the large intestine.Dd: Trichuriasis. which may vary from watery to mucoid. Porcine intestinal spirochaetosis (spirochaetal diarrhoea) Porcine intestinal spirochaetosis appears to be a distinct entity associated with infection by Brachyspira (formerly Serpulina) pilosicoli. iii. gastric ulceration) Specimens required i. confirmation of T suis by parasitological examination. in one associated with a pelleted diet. Histological changes comprise hypertrophy of the colonic mucosa. It is a diarrhoeal disease of unknown aetiology. elimination of other possible causes of ill thrift (e. resolving in 7-10 days. or mild thickening and oedema of the colonic mucosa with localised mucosal haemorrhages. mainly lymphocytic. or Lawsonia intracellularis. Diagnosis is confirmed by assessment of the diet. comprising epithelial cell loss. marked weight loss. the only change detected is frothy large intestinal contents. or becoming chronic. Yersinia spp). Lesions are often mild. yersiniosis. gastric ulcers. and a thickening of the colonic mucosa. bacteriology. patchy and focal superficial colitis and possibly typhlitis. Diarrhoea may progress from ‘wet cement’ or ‘cow pat’ faeces to watery diarrhoea with dehydration. poor quality oils and carbohydrates. and leads to loose (’wet cement) faeces. . Disease outbreaks can be caused by massive infestations of immature T suis (no T suis ova in faeces). In resolving lesions. with corn as the major component. Diagnosis Clinical signs (weaner ill-thrift associated with diarrhoea or mucoid faeces). and B pilosicoli may be demonstrated in affected mucosa by PCR or culture. post-weaning colibacillosis. with shallow. and VETERINARY LABORATORY MANUAL replacement of lost epithelium by low cuboidal enterocytes. Dietary niacin deficiency Dietary niacin deficiency produces severe. submitted chilled for bacteriology and PCR for differential diagnosis. conical scales of fibrinonecrotic material may be dislodged from the mucosal surface by rinsing. ii. There may be no gross lesions.g. persistent diarrhoea. Confirmation of intestinal spirochaetosis due to B pilosicoli. histopathology. postmortem findings.parasitology. B hyodysenteriae. infiltration of the lamina propria. iv. reduction of goblet cell numbers. which are not as easily detected at necropsy as the adult worms. Impression smears of affected intestinal mucosa for bacteriology. Histological changes include mucosal oedema. Salmonella spp. Sections of affected intestine. submitted in buffered formalin for histopathology. spirochaetes may cover an intact epithelium as a false ‘brush border’. porcine proliferative enteropathy (Lawsonia intracellularis infection) Porcine colitis has been attributed to various non-Brachyspira hyodysenteriae spirochaetes and Campylobacter coli. especially those 8-14 weeks of age. Lesions include proprial leucocytosis. with variable epithelial necrosis and erosion. It is likely that this syndrome has a variety of causes. Many cases reported previously may have been undiagnosed infections by B pilosicoli or possibly other weakly beta-haemolytic spirochaetes. E coli. It is possible that high energy grower diets may escape complete digestion in the small intestine.

Yeerongpilly. ii.NB. PORCINE PLEUROPNEUMONIA Actinobacillus pleuropneumoniae (formerly Haemophilus pleuropneumoniae) causes a fatal pleuropneumonia or non fatal chronic respiratory disease. Fixed affected lung in buffered formalin for histopathology. Nasal swab material collected into sterile PBS for PCR (if required. oedema disease. Serum (or heparin plasma) samples from affected pigs and their cohorts for ELISA. Concurrent infection with Pasteurella multocida is not uncommon. and dyspnoea. ii. Culture is expensive and not routinely available. Qld. PORCINE ENTEROVIRUS ENCEPHALOMYELITIS Syn: Talfan disease A sporadic encephalomyelitis usually affecting pigs from 14 days to weaning. Specimens required i. torsion of mesentery. colonic bloat. hyopneumoniae infection status of the herd. 11. Clinical signs include abdominal distension and pain. PORCINE ENZOOTIC PNEUMONIA Syn: Mycoplasmal pneumonia Diagnosis Identification of Mycoplasma hyopneumoniae infection in a herd by ELISA serology. Diagnosis History. ii. whey feeding. samples taken from the affected site and measuring 1 cm³ is sufficient. 2. Brain fixed in neutral buffered formalin for histopathology. Brachyspira pilosicoli and Lawsonia intracellularis. but also seen in older pigs. Predisposing factors include transportation. Carriage in the live animal can be detected by PCR from nasal swabs. However. clinical signs. Fixed affected lung for histopathology iii. 3. ELISA serology can provide a profile of the M.g. fixed spinal cord for histopathology. 7. Seroconversion after natural challenge can take 9 weeks. iii. Diagnosis Gross pathology. or in individual animals by PCR on affected lung. but most cases are simply found 91 . Focal and diffuse gross lesions of reddish black colour with or without pleurisy are suggestive. Since this is a herd disease. Intestinal volvulus. Fresh lung samples submitted chilled or frozen for PCR. Sample of fresh. these must be fresh and reach the laboratory within a few hrs of slaughter.g. for intensive investigation or research). For this reason. with recumbency and paddling. Other samples to eliminate possible differential diagnoses: e. Diagnosis Histopathology. Fresh affected lung and bronchial lymph node for culture. 12) occur in Australia isolates are currently being serotyped at the Animal Research Institute. 5. Blood samples may be collected from the abattoir or from a subpopulation of animals in the piggery. Typically causes posterior paresis and ataxia. redgut. At least 7 serotypes (1.including brain and small intestine for bacteriology. bacteriology and histopathology. nasal excretion is variable. ii. so a negative swab PCR on a single occasion is of limited diagnostic value. spinal cord abscess or other lesions . post-mortem findings. For samples where an isolate is required (e. PCR tests are available for three enteric pathogens: Brachyspira hyodysenteriae. Tremors and excitement may occur. Specimens required i. intestinal venous infarction. chilled brain for virus isolation. Low morbidity and mortality. looked at in this light rather than on an individual animal basis. For PCR. virus isolation. whey bloat. the results should be VETERINARY LABORATORY MANUAL PORCINE INTESTINAL HAEMORRHAGE SYNDROME Syn. may progress to all legs. Specimens required Live pigs: i. may be cost prohibitive) Slaughter pigs: i. and preferably come from well developed lesions. bacterial meningoencephalitis. so targeting older pigs for serological profiling is more cost-efficient. it is recommended that about 30-40 sera be submitted on each occasion.

intracellularis lesion. and bacteriological examination. while additional changes that may be superimposed on this lesion may result in regional ileitis (muscular hypertrophy). ii. lung and heart blood submitted chilled for bacteriology. PCR tests are available for three enteric pathogens: Brachyspira hyodysenteriae. necrotic enteritis Diagnosis Clinical signs. consistent with congestive heart failure. kidney. histopathology. Sections of affected intestine. lung. virology and histopathology. spleen. Syn: Porcine intestinal adenomatosis (PIA). liver. Fees for tests undertaken to confirm or exclude a diagnosis of porcine myocarditis are paid by NSW Department of Primary Industries. vi. ileitis. PORCINE PROLIFERATIVE ENTEROPATHY Porcine Proliferative Enteropathy VETERINARY LABORATORY MANUAL 92 . There may be a variable increase in the incidence of mummified foetuses. submitted chilled for virology. Chilled stillborn piglets for necropsy. occurring in utero. postmortem findings.gov. regional ileitis (RI). Sections of affected intestine (especially ileum and colon). Impression smears of affected intestinal mucosa for bacteriology (MZN stain) iv. . Sections of liver.dead. submitted in buffered formalin for histopathology iii. kidney. iv. which receives its blood supply from the gastroduodenal vessels. PMC is a notifiable disease. Usually but not always associated with palpable torsion of the root of the mesentery that causes intestinal venous infarction.htm. Diagnosis History. PMC causes an increase in stillbirths and pre-weaning mortalities. histopathological examination of affected intestine. characterised by a susceptibility to stress-induced collapse and death.au/reader/anhealth/pmc-qa. There is often evidence of cardiac enlargement and an increase in the volume of body fluids. Lawsonia intracellularis is the aetiological agent. Specimens required i. PORCINE MYOCARDITIS (PMC) Porcine myocarditis (PMC) is a recently recognised condition of uncertain aetiology apparently affecting pigs in only two piggeries in NSW. Brachyspira pilosicoli and Lawsonia intracellularis. PHE is associated with sudden death in finisher and breeder pigs. but PIA can occur in all ages. is unaffected. clinical signs. It is believed that the syndrome is due to a viral infection predominantly. particularly of Landrace and Pietrain breeds. submitted chilled for bacteriology for differential diagnosis. Fresh heart. necropsy findings. necrotic enteritis (coagulative necrosis) or PHE (haemorrhage). Specimens required iii. The uncomplicated proliferation of mucosa due to L. Sections of heart.agric. spleen and whole brain in buffered formalin for histopathology. PIA and RI are associated with weaner ill thrift. Histologically there is a non-suppurative myocarditis Further information can be obtained from the NSW Department of Primary Industries’ website at http://www. It is likely that necrotic enteritis represents secondary bacterial infection superimposed on a primary L. It is an intracellular organism and requires rat enterocyte cell lines for maintenance. if not exclusively. while necrotic enteritis can cause mortalities in grower pigs. v. proliferative haemorrhagic enteropathy (PHE). The proximal duodenum. PORCINE STRESS SYNDROME (PSS) Syn: Pale soft exudative pork [PSE]. malignant hyperthermia An inherited neuromuscular disease of grower pigs. Fresh intestinal contents or faeces for PCR NB. lung and body fluid collected separately and aseptically. The gross pathological changes consist of small pale areas in the myocardium.nsw. intracellularis is referred to as PIA.

It is caused by a recessive mutation of the HAL gene on chromosome 6 (susceptibility to PSS was initially determined by reaction to the anaesthetic halothane). Animals can be genotyped as stress-resistant (NN), stress-carrier (Nn), or stress-positive (PSSpositive) (nn). A large proportion of the stress-carrier and PSS-positive animals produce carcases with inferior muscle quality. Diagnosis Clinical history and signs, post mortem findings (60-70% of pigs may show gross muscle lesions). Laboratory examination of necropsy specimens cannot assist the field diagnosis. A DNA test for PSS genotyping patented by the University of Toronto is available at commercial laboratories in North America. It is not routinely available in Australia.

Specimens required See http://www.dpi.nsw.gov.au/agriculture/vetm anual/specimens-by-diseasesyndrome/diseases_of_livestock/dpi-sampcoll-prot.pdf Diagnosis of the disease and heterozygote detection i. 20 to 25 hairs, with roots attached, from the distal end of the tail.

PYELONEPHRITIS (BOVINE)
Diagnosis Clinical signs mainly of abnormal urine and recovery of Corynebacterium renale from urine or kidney tissue. Specimens required i. Sample of urine collected into a sterile container, submitted chilled for bacteriology. ii. Sections of kidney lesion submitted chilled for bacteriology. iii. Sections of kidney and other parts of urogenital tract with lesions, submitted in buffered formalin for histopathology.

PREGNANCY TOXAEMIA
Diagnosis Generally on clinical grounds and demonstration of ketones in urine. Differentiate from hypocalcaemia, metabolic or starvation ketosis, secondary ketosis due to anorexia from other causes and polioencephalomalacia. Specimens required Samples are only to possible causes: i.

PYOGENIC INFECTIONS
Diagnosis Recovery of pyogenic organisms from abscesses and infected organs. Specimens required i. At least four (4) smears from periphery of the lesion for bacteriology. ii. Portions of affected tissue containing lesions, submitted chilled for bacteriology. iii. Section of lesion, including the margin, submitted in buffered formalin for histopathology.

eliminate

other

ii.

Sections of liver, kidney, adrenal gland and brain submitted in buffered formalin for histopathology. At least 2 ml of serum, free of cells and haemolysis, submitted chilled for calcium, magnesium and beta hydroxybutyrate (ketone) estimation.

NB Urine examination for ketones should be carried out in the field.

PSEUDOCOWPOX
See Teat lesions

PYRROLIZIDINE ALKALOIDOSIS
See also Poisoning, plant' An intoxication of grazing sheep, cattle, pigs and horses when hepatotoxic pyrrolizidine alkaloids are ingested from plants of the genera Senecio, Crotalaria, Heliotropium, Amsinckia, Echium and others. In sheep, the condition may be complicated by chronic copper poisoning. Diagnosis History of access to hepatotoxic plants, clinical and post mortem findings, histopathology.

PROTOPORPHYRIA
(Limousin and Blonde d'Aquitaine cattle) An acute photosensitivity evident from birth and characterised by alopecia, ulceration and scarring of the pinnae, nares and possibly along the midline. Diagnosis Clinical signs and history; confirmed by DNA analysis.
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93

Specimens required i. Sections of liver and kidney, submitted in buffered formalin for histopathology. ii. In cases of suspected copper poisoning, at least 30 g of liver and kidney, in separate copper free jars, submitted chilled for copper estimations.

RHODOCOCCUS EQUI PNEUMONIA IN HORSES
Syn: Rattles Diagnosis History, clinical signs, and post mortem findings. Recovery of Rhodococcus equi from lungs and pulmonary lymph nodes. Specimens required i. Portions of affected lung and lymph nodes submitted chilled for bacteriology. ii. Swabs of lung abscesses in Amies Charcoal Transport medium, submitted chilled for bacteriology. iii. Portion of affected lung submitted fixed in buffered formalin for histopathology.

Q FEVER
Q Fever is usually an asymptomatic infection of animals caused by Coxiella burnetii, but it can cause abortion in sheep and goats and infections in man. Diagnosis The complement fixation test is not a reliable test for Q Fever. At present there is no routine test available for diagnosis. The CFT is used only for export certification. Specimens required i. For abortions, those required for abortion in sheep and goats. ii. For export certification, serum sample forwarded chilled for CFT.

RHODOCOCCUS EQUI LYMPHADENITIS IN PIGS
Characterised by oval or spherical lesions in lymph nodes which are encapsulated and easily enucleated. It is often difficult to differentiate from tuberculosis in swine. Diagnosis Recovery of Rhodococcus lesions, histopathology.

RED GUT IN SHEEP
Syn: Intestinal volvulus, torsion of mesentery, colonic bloat, intestinal venous infarction An acute haemorrhagic enterocolitis occurring in sheep grazing some lucerne or clover pastures, or other fresh, young green feed. Some cases show severe abdominal distension, with rapid death. Post mortem findings can include distension of the small and large intestine, severe haemorrhage and congestion of intestinal mucosa and blood stained intestinal contents. There may be rapid autolysis of the carcase. Diagnosis Post mortem findings, especially displacement and/or torsion of the caecum and colon. Differential diagnosis from enterotoxaemia. NB. The ventral necropsy approach is essential for the displacement and/or torsion to be seen. Specimens required i. As for enterotoxaemia, together with ii. Sections of affected intestine, submitted in buffered formalin for histopathology.

equi

from

Specimens required i. Affected lymph nodes, submitted chilled for bacteriology. ii. Affected lymph nodes, in buffered formalin for histopathology.

RINGWORM (DERMATOMYCOSIS)
See fungal infections

ROCK FERN POISONING
Cheilanthes tenuifolia (rock fern) can cause haemorrhages through out the body with hepatic necrosis. Also associated with staggers or ataxia in sheep. Diagnosis History, clinical signs, post mortem findings, histopathology. Specimens required Live animal i. 5 ml of blood in EDTA for haematology. ii. Blood films for haematology (refer to Haematology in first section of this manual). Dead animal

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94

i.

ii.

Sections of liver, spleen and kidney submitted in buffered formalin for histopathology. Specimens to differentiate from other causes of haemorrhage and septicaemia, based on post mortem findings.

Diagnosis Clinical signs, necropsy findings, recovery of Salmonella from heart blood, spleen, liver, bile, mesenteric lymph nodes and intestinal contents in both septicaemic and acute forms. Serology. In the chronic form, the bacteria may be recovered from the intestinal lesions and less frequently from other viscera. Specimens required Live animal i. Serum for Salmonella Typhimurium and Salmonella Dublin SAT serology in cattle and sheep. ii. Faecal sample submitted chilled for bacteriology. Dead animal: i. Portion of liver, kidney, lung, bile, spleen, mesenteric lymph nodes and small intestinal contents in separate sterile containers, submitted chilled for bacteriology. ii. Sections of liver, kidney, lung, spleen and small and large intestine, submitted in buffered formalin for histopathology.

Ataxic animal Specimens as required for Ataxia.

ROTAVIRUS INFECTION
See also Scouring Causes diarrhoea in young animals. Diagnosis Demonstration of either virus particles in faeces by electron microscopy or soluble antigen by immunodiffusion, latex agglutination or ELISA. Specimens required i. 20 g of faeces or intestinal contents, submitted chilled for electron microscopy or antigen detection. ii. Specimens as required for the differential diagnosis of causes of scouring.

RYEGRASS STAGGERS
Syn; Perennial ryegrass staggers Associated with fungal neurotoxins produced by Acremonium loliae in the leaf, stem and seed of perennial ryegrass, which are toxic to sheep and cattle. Outbreaks more common in summer and autumn, and with close grazing of limited available feed. Diagnosis History, clinical signs, histopathology. This syndrome can be diagnosed by clinical signs in the field. Specimens required i. Brain, submitted in buffered formalin for histopathology.

SALT POISONING IN PIGS
Diagnosis History, clinical signs, histopathology. Specimens required i. Brain, submitted in buffered formalin for histopathology. ii. Specimens as required for the differential diagnosis of causes of nervous disorders.

SARCOSPORIDIOSIS
Infection with protozoa of the genus Sarcocystis can cause anaemia, ill thrift and myopathy in experimental infections in sheep and an acute febrile disease in calves. Diagnosis Histopathology. Specimens required i. Sections of cardiac and skeletal muscle, submitted in buffered formalin for histopathology.

SALMONELLOSIS
Usually occurs as one of three major syndromes: • Peracute septicaemia seen mostly in young animals. • Acute enteric form most commonly seen in adult animals. • Chronic enteric form is common in pigs, and occurs in cattle. There is persistent diarrhoea, severe emaciation and intermittent fever.
VETERINARY LABORATORY MANUAL

SCABBY MOUTH
See Contagious pustular dermatitis'

95

SCOURING Investigations into causes of scouring in animals should take into consideration: • Age of affected animal9s) • Whether sudden or gradual in onset • Condition of the animal • Numbers affected • Intensity of grazing or housing Diagnosis History. bacteriology. For live/dead sperm examination. In cases of suspected enteric or systemic infections at least 30 g of faeces. Veterinarians should conduct the initial examination in the field. iv. or • There is evidence of inflammatory products in the sample. nutritional' SEMEN EXAMINATION In investigations of infertility. dogs associated with Campylobacter jejuni • Copper or selenium deficiency in ruminants • Coccidiosis • Hypomagnesaemia in milk fed calves In general. especially areas showing lesions. polymorphs. virology. For morphology. ruminant metabolic profile. arsenic poisoning. spleen. renal profile. submitted chilled for bacteriology. ii. Specimens as required for specific diseases. Sections of liver. especially in neonates • Rotavirus infection in young animals • Cryptosporidiosis in young animals • Salmonellosis • Pestivirus (Mucosal disease) in cattle. ii. Test results are incorporated into the laboratory report which includes normal ranges supplied by the testing laboratory. Dead animal: i.e. clotted semen. if these are suspected. Sections of small and large intestine. iii. Johne's disease. submitted chilled for bacteriology or virology. sheep and goats • Yersiniosis • Poisoning. parasitology. Pestivirus. Specimens required Depending on the age of the animal and the results of the clinical examination. histopathology. kidney. wave motion. especially < 2 yrs • Parasitism • Johne's disease in cattle. the following specimens should be submitted.g. and • There is an obvious deficiency with regard to density. For bacteriology. At least 30 g of faeces from 10 to 20 animals in the flock. i. ii. selected sites in the gastrointestinal VETERINARY LABORATORY MANUAL tract. semen collected aseptically and submitted in sterile containers. specimens should be forwarded as required for the following diseases: • Colibacillosis. plant or chemical • Proliferative enteropathy in pigs • Swine dysentery in pigs • Enteritis in pigs. Serum enzymology is usually undertaken as part of a package of tests eg liver profile. cardiac and skeletal muscle in buffered formalin for histopathology. Live animal i. Sections of liver. motility. submitted chilled for parasitology. v. The most frequently requested serum enzymes are: • Aspartate aminotransferase (AST) also known as Glutamic oxaloacetic transaminase (GOT) 96 . semen examination should be considered after a careful clinical examination. SERUM ENZYMOLOGY NSW Department of Primary Industries refers all clinical chemistry testing to a specialist laboratory. iii. lung and spleen submitted chilled for bacteriology in cases of suspected enteric or systemic infection. Abomasum and small intestine submitted chilled for total worm count. clinical examination. Blood or serum samples as required for virology. calves. iii. NB. Laboratory examination can be considered: • When the field veterinarian is confident the semen sample is typical of the semen to be obtained. e. Semen collected using an artificial vagina is not suitable for bacteriology. SELENIUM DEFICIENCY See Muscular degeneration. kidney. smears stained with freshly prepared nigrosin eosin. Specimens required i. air dried semen smears.

clinical serology. PE) GGT Bile duct epithelium damage. Specimens required i. ii.g. Brain and sections of liver. pyrrolizidine alkaloidosis) or acute hepatic necrosis GLDH Hepatocellular damage (rises rapidly in acute hepatic necrosis) SODIUM FLUOROACETATE (1080) POISONING Causes neurological signs in canines and cardiac signs in ruminants. signs. It must be free of haemolysis. Diagnosis History.. Specimens will be sent to a specialist laboratory on request. chronic fascioliasis. of stomach poisoning in sections of and lung Specimens required i. detection of poisoned material. 97 . kidney and other organs with serositis. e. CNS < 300 GGT Liver < 80 sheep < 50 cattle GLDH Liver < 20 sheep < 50 cattle Typical disease syndromes with elevation of serum enzymes Enzyme Disease process AST Sustained necrosis of hepatic and/or muscular tissue (skeletal or cardiac) CK Sustained necrosis of muscular tissue (eg. ii. SPORADIC BOVINE ENCEPHALOMYELITIS (SBE) See also Chlamydial infections' Diagnosis History. Paired serum samples from individually identified and observed animals collected in the clinical and convalescent phase (2 to 3 weeks apart). it should be frozen if the delay will be greater than 24 hrs. submitted in buffered formalin for histopathology. Continued exposure to low doses can cause myocardial degeneration in cattle and sheep. spleen. iii. NSW Department of Primary Industries does not analyse for fluoroacetate. liver VETERINARY LABORATORY MANUAL submitted in buffered formalin for histopathology. Approximately 100ml contents. elimination of other causes. clotted in vacuum tubes. submitted chilled or frozen for chlamydial serology. chemical analysis confirms fluoroacetate in stomach contents. submitted chilled for bacteriology. muscle < 100 CK Muscle. WMD) and/or CNS (eg. NB If blood cannot be sent to the laboratory overnight. myocardium. cholestasis incl cases with biliary hyperplasia (eg. carrot in the stomach contents. as is their half life in the blood: Stability in serum sample Intermediate Unstable Very stable Intermediate Half life in blood after insult Weeks Days Weeks AST CK GGT GLDH Applications and Interpretation of Results in Sheep and Cattle Enzyme Major source Suggested normal serum levels (IU/L) AST Liver. Note that the stability of these enzymes is variable. 10 ml of blood. Approximately 2ml of serum per animal should be provided. also known as creatine phosphokinase (CPK) Gamma glutamyltransferase (GGT) Glutamate dehydrogenase (GLDH) Specimens required i. histopathology. In cases of suspected sheep and cattle. Portions of liver and spleen collected aseptically.• • • Creatine kinase (CK). it should be allowed to clot and the serum poured off.

Swine dysentery may be difficult to detect in straw-based shelters. spleen ii. Brachyspira hyodysenteriae (formerly Serpulina hyodysenteriae) is a relatively common pathogen with a severe impact on production. lung. liver. with blood and mucus. spleen. SWINE DYSENTERY Syn: Brachyspira hyodysenteriae infectionDD: Lawsonia intracellularis (proliferative enteritis/ileitis). Diagnosis History of access to P simplex. and affected pigs can vary from the classical mucohaemorrhagic diarrhoea . liver. There are no laboratory fees for tests taken to exclude strangles. Specimens required i. grey to light brown to dark. isolation of Streptococcus equi from lesions. Diagnosis Isolation of S suis from affected tissues (brain. kidney. ST GEORGE DISEASE Caused by ingestion of desert riceflower (Pimelea simplex). kidney. less commonly: Salmonella. Brachyspira pilosicoli (intestinal spirochaetosis. Pus samples or swabs taken from lesions and upper respiratory tract. STREPTOCOCCUS SUIS INFECTION IN PIGS Streptococcus suis is associated with meningitis. Sections of liver.iv. Submission of appropriate specimens accompanied by a Specimen Submission Form requesting exclusion of strangles is suitable notification. SWAINSONA POISONING See also Poisoning.3. There is gradual spread in group. Swine dysentery can occur in any age (suckers . Specimens required i. submitted in buffered formalin for histopathology. kidney in buffered formalin for histopathology Serotyping of S suis isolates is not routinely available. Fresh chilled portions of brain and/or lung. or faeces varying from soft. spleen). histopathology. plant' Diagnosis History. submitted chilled in Amies charcoal transport medium for bacteriology. Approximately 100ml of stomach contents. At least 28 serotypes have been identified. and stained hindquarters.7 and 9. kidney and lymph nodes.2. Heparinised or citrated blood from acute cases for chlamydial isolation and FAT. dark faeces ("black scour") is associated with intermittent medication. kidney. Specimens will be sent to a specialist laboratory on request.breeders) but is most common in 15-70 kg grower pigs (5-20 weeks). chronic form of loose. clinical signs. History is typically one of depressed growth rate. confirmation by detection of strychnine in ingesta by chemical analysis. Gastric ulcers. Clinical expression is normally low mortality/ high morbidity. and increased incidence of "slab 98 STRYCHNINE POISONING Diagnosis VETERINARY LABORATORY MANUAL . endocarditis and bronchopneumonia in young pigs.various amounts blood and mucus. A less severe. STRANGLES IN HORSES Strangles is a notifiable diseases in NSW. heart and lung. liver. Disease in Australian pigs has been associated with serotypes 1. septicaemia. joints. Brain and spinal cord. Sections of liver. Specimens required i. ii. Sections of brain. NSW Department of Primary Industries does not analyse for strychnine. Diagnosis Clinical findings. Clinical signs. polyarthritis. Whipworm (Trichuriasis). spirochaetal colitis). Specimens required i. submitted in buffered formalin for histopathology. clinical signs. Severe cases can show watery faeces. submitted in buffered formalin for histopathology. and partial loss of appetite that further decreases growth. Specimens required i. Non-specific (dietary) colitis. It may be less common in weaners if medication for respiratory diseases is used.

Fresh. Smears of affected intestinal mucosa. Specimens should be forwarded to eliminate other causes. Specimens required Live animal i. linked to a history of ‘ordinary’ growth rate. NB: PCR are available for Lawsonia intracellularis and Brachyspira pilosicoli. Serology is available to exclude Lawsonia. bovine herpes mammilitis virus. Epithelial hyperplasia is a feature. Diagnosis Response to treatment. demonstration of organisms in blood or organ smears. Dead pigs i. bacteriology. Abattoir inspection may reveal an index case through evidence of an accumulation of blood-tinged and jelly-like material in the large intestinal wall and between the coils. Submission of appropriate specimens accompanied by a Specimen Submission Form requesting exclusion of tick fever is suitable notification. if clinical signs have been evident for over one week. with erosions. Conventional unstained thin blood smears for haematology. differentiated from other scouring. focal symmetrical encephalomalacia. This is often severe and extensive. Specimens required i. Babesia bovis (argentina) infections often cause nervous symptoms. followed by coma and death. strychnine poisoning. chapping and photosensitisation. avoiding exposure to heat or direct sunlight. jaundice and anaemia. ii. demonstration of virus by electron microscopy. There are no laboratory fees for tests taken to exclude tick fever. causing a fibrinohaemorrhagic colitis and typhlitis. . fresh blood and mucus on necropsy. Specimens required Live pigs i. and necrotic debris chilled for virology (electron microscopy examination) and bacteriology. Fixed affected intestinal section in buffered formalin for histopathology Can be caused by infection by paravaccinia virus (pseudocowpox).g. Thick blood films air dried. Culture of affected intestine or faeces. bacterial infections. for parasitology. Specimens required The condition should be diagnosed in the field. clinical signs. ii. Identification of Brachyspira hyodysenteriae infection in individual animals by PCR on affected intestine or faeces. ii. herpes mammilitis virus' VETERINARY LABORATORY MANUAL 99 . Diagnosis Clinical signs. for antibiotic sensitivity). must causes be of TEAT LESIONS Includes Pseudocowpox. elimination of other causes including polioencephalomalacia. Scabs with underlying tissue. Diagnosis Clinical signs. Pathology affects proximal large intestine. Faeces for PCR ii. In the Tick Quarantine Area. Biopsy of lesion in buffered formalin for histopathology TETANUS Diagnosis History. chilled portion of affected gut for smears. elevated temperature.sided" pigs (hollow in flanks). (see Specimens for Haematology). Specimens required There is no routine diagnostic test for tetanus. TAPEWORM INFESTATION IN SHEEP Occasionally suspected of causing ill thrift and scouring in sheep under poor nutritional conditions. PCR and possible culture. Faeces for culture if isolation is required (e. It is preferable to advise the laboratory so special selective media can be available. tick fever should be suspected in animals showing haemoglobinuria. Diagnosis Gross and histopathology. TICK FEVER OF CATTLE Includes Anaplasmosis. babesiosis Tick fevers are notifiable diseases in NSW.

or • Transplacentally. Serological tests are not reliable in cats. Diagnosis Typical lesions in placenta and foetal brain (necrogranulomas in brain). heart. NB Foetal membranes must be submitted if toxoplasmosis is to be diagnosed. submitted chilled for bacteriology and parasitology. The asexual proliferative stages of Toxoplasma gondii with formation of tissue cysts. liver. submitted in buffered formalin for histopathology. heart. Oocysts of Toxoplasma gondii are highly infectious to humans and animals. v. liver. v. iv. Diagnosis Clinical signs. muscle. In instances where clinical disease requires confirmation in the dead animal. Sections of liver. or by T gondii cysts in infected intermediate-host tissues (eg infected placenta). muscle and brain. iii. Clinical disease is not common in cats.iii. TOXOPLASMOSIS IN SHEEP AND GOATS Abortion and perinatal loss in both sheep and goats is caused by the asexual proliferative stages of Toxoplasma gondii. with sexual stages in the gut resulting in oocysts shed in faeces. submitted in buffered formalin for histopathology. air dried for parasitology. heart. Serology by Latex test. submitted chilled for parasitology and haematology. Serological titres of 4 or more in the Latex test are considered positive. Sections of kidney. kidney. risk to laboratory workers. Impression smears from the kidney. with formation of tissue cysts. Infection of the dam can be by sporulated T gondii oocysts from cats. • Consuming tissues of other intermediate hosts containing T gondii cysts. Urine sample. Blood collected into EDTA. Sections of kidney. and brain should be submitted for histopathology. Specimens required i. At least 2 ml of serum. • Consuming tissues of other intermediate hosts containing T gondii cysts. Blood films. brain and spleen. heart. prepared by crushing a section of the cerebral grey matter the size of a match head between 2 slides and spreading lightly. oocyst excretion is sporadic. ii. and in small numbers. Those required for abortion in sheep (see 'Abortion in Sheep'). Except initially. or • Transplacentally. heart. TOXAEMIC JAUNDICE See Copper poisoning TOXOPLASMOSIS IN CATS The cat is the definitive host of the coccidian parasite Toxoplasma gondii. 100 . iv. There are no routine serological tests available for animals other than sheep and goats. kidney. cause disease in a wide range of intermediate animal hosts (including humans) that are infected by • Consuming sporulated T gondii oocysts shed by cats. histopathology. fixed sections in buffered formalin of small intestine. cause disease in a wide range of intermediate animal hosts (including humans) that are infected by • Consuming sporulated T gondii oocysts shed by cats. Faecal examination will only be performed in special circumstances after prior arrangements have been made with the laboratory. spleen and brain. Dead animal i. liver and brain for parasitology. spleen. Such examinations pose a high VETERINARY LABORATORY MANUAL TOXOPLASMOSIS IN OTHER ANIMALS The asexual proliferative stages of Toxoplasma gondii with formation of tissue cysts. spleen. There is no routine serological test available. submitted chilled for bacteriology and parasitology. Brain squash preparation from the grey matter of the cerebral cortex. submitted chilled for serology. Specimens required i. liver.

Submission of appropriate specimens accompanied by a Specimen Submission Form requesting exclusion of tuberculosis is suitable notification. Herd infertility i. See 'Abortion in Cattle'.gov. Brain and cord in buffered formalin for histopathology. There are no laboratory fees for tests taken to exclude tuberculosis. vaginal or uterine exudates of vaginal mucus from selected females taken 10 to 20 days after service submitted in selective transport medium for T. Specimens required i. Specimens as required for abortion in cattle. tuberculin test. scrapie in sheep and goats The National TSE Surveillance Program (NTSESP) (http://www. caused by the protozoon Tritrichomonas foetus. recognition and/or recovery of pathogenic mycobacteria from tissues.au/agriculture/vetm anual/specimens-bydiscipline/bacteriology/02-bull-vd-coll. freeze fresh tissues (and prevent from thawing during transport). See http://www. histopathology. If considerable delays (> 3 d) will occur before culture.au/a ahc/programs/adsp/tsefap/tsefap_home. necropsy findings. ii. Specimens required Suspect TB lesions i. VETERINARY LABORATORY MANUAL 101 . chilled cervical spinal cord* for detection of prion protein.au/agricultur e/vetmanual/specimens-bydiscipline/bacteriology/02-in-pouchguide.nsw. ii. Portions of lesions unpreserved in sterile sealed jars.dpi. unfixed cervical spinal from all cases of progressive neurological disease in any animal species. for testing for PrP if required.pdf TUBERCULOSIS Diagnosis Clinical examination.cf m) is a national program jointly funded by government and industry to demonstrate Australia’s ongoing freedom from BSE and scrapie. histopathology. Diagnosis Herd history of infertility and abortion.gov. bovine spongiform encephalopathy (BSE). and to provide early detection of those diseases should they occur. See http://www. preputial scrapings submitted unchilled in selective transport medium for T foetus (InPouch TF). When required. Fresh. From bulls. The NTSESP provides payments for producers. submitted chilled for bacteriology. Specimens required Abortion i.pdf NB Selective transport medium for T. • A completed Specimen Submission Form Diagnosis Clinical signs of progressive neurological disease. There are no laboratory fees for tests taken to exclude trichomoniasis.animalhealthaustralia. usually in the brain and cord. From cows. Trichomoniasis is a notifiable disease in NSW. demonstration of T foetus. NTSESP submissions must be accompanied by: • A completed NTSESP Clinical History and Post-mortem Report Form. *It is prudent to routinely collect at least a sample of fresh. private veterinarians and government agencies for the submission of specimens for laboratory testing from adult cattle and sheep with progressive neurological disease.com.nsw.TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY (TSE) Syn: Prion disease. specimens for culture of mycobacteria will be submitted to the Australian Reference Laboratory in Western Australia.dpi. Immunochemistry is used to specifically identify accumulated abnormal prion protein (PrP). Submission of appropriate specimens accompanied by a Specimen Submission Form requesting exclusion of trichomoniasis is suitable notification. abortion and pyometron. TRICHOMONIASIS OF CATTLE A contagious venereal disease of cattle causing infertility. foetus (InPouch TF) together with instructions for the collection and despatch of specimens will be forwarded from the laboratory upon request. Tuberculosis is a notifiable disease in NSW. foetus.

clinical signs of ataxia. Portions of lesions. and submitted chilled for culture: Essential • L & R medial retropharyngeal LN (also known as suprapharyngeal LN) • L & R bronchial LN • Cranial and caudal mediastinal LN • L & R Supramammary LN (in cows) Desirable • L & R lateral retropharyngeal LN (also known as atlantal LN) • L & R mandibular LN • L & R parotid LN • Mesenteric LN • L & R internal iliac LN It is important that the property of origin be clearly identified. encephalitis. coma. . Ad lib feeding. NB. NB. In the case of tuberculin reactor cattle. Specimens required i. • diabetes • brain lesions including tumours. UREA POISONING Diagnosis History of access to urea. care should be taken in collecting samples. ii. The field veterinarian should then consider diagnostic possibilities according to the results of the urine examination and make a field diagnosis or send the appropriate specimens. Diagnosis Gross pathology is usually sufficient to confirm the diagnosis. As organisms responsible for tuberculosis in livestock are pathogenic for man. haemorrhage. Specimens should be forwarded in such a manner that they will offer no risk of infection to persons handling the containers in transit or on arrival at the laboratory. Tissues are cultured for 12 weeks before being classed as negative. especially if due to chemical poisoning • inflammation of the postrenal urinary tract • chronic passive congestion of the kidney due to a number of conditions including cardiac and hepatic diseases and fevers. A biopsy from the affected area. Glycosuria Large quantities of ketone bodies may depress the colour development and result in a false negative reaction. Specimens required There are no routine laboratory procedures available to diagnose urea poisoning. Reactor cattle i. either by name or tail tag. or tattoo in the case of pigs. also sometimes in listeriosis and botulism Acetonuria (Ketonuria) VETERINARY LABORATORY MANUAL 102 ULCERATIVE SPIROCHAETOSIS IN PIGS Diagnosis Clinical signs. Positive cultures may appear at approx 8 weeks (possibly earlier in lesioned tissues with massive infection). in buffered formalin for histopathology. Smears from skin lesions. the following lymph nodes should be collected as aseptically as possible in separate sterile containers. URINE EXAMINATION Urine examinations should be performed in the field using the commercially available dip stick kits.ii. The results obtained should be interpreted according to the instructions with the kit. respiratory distress. stress and finely ground grains increase the prevalence of lesions. • enterotoxaemia in sheep. histopathology. Suspect tuberculosis specimens must be packed and despatched separately from other specimens. submitted in buffered formalin for histopathology. and can be confirmed either as Mycobacterium bovis or not Mycobacterium bovis by monoclonal dot blot in a matter of days. ULCERS. providing there are sufficient colonies for testing. Proteinuria • following violent exercise or stress • nephritis • haematuria from any cause • nephrosis. GASTRIC IN PIGS Haemorrhage from an ulcerated of the pars oesophagea with haemorrhage is an important cause of anaemia and sudden death in grower and finisher pigs. bacteriology.

Y intermedia. NSW Department of Primary Industries laboratories do not analyse for vitamin A or vitamin E. lactation. Syndromes associated with yersiniosis Y pseudotuberculosis and Y enterocolitica in domestic animals: 103 .Occurs in any clinical condition associated with deficient carbohydrate metabolism. Interpretation of vitamin A and E concentrations in blood and liver (cattle. pools may be prepared where possible. haemoglobinuria • neoplasia of the urinary tract • urolithiasis • infectious diseases. abortion and perinatal mortality Pneumonia Mastitis Epididymo-orchitis In addition. Y kristenseni and Y frederickseni occasionally cause disease.9 < 4.6 Pig (piglet) Liver µmol/kg < 11 <7 Pig (adult) Liver µmol/kg < 40 <7 VOMITING AND WASTING DISEASE OF PIGS See Haemagglutinating encephalomyelitis virus (HEV) infection' WHITE MUSCLE DISEASE See Muscular degeneration nutritional YERSINIOSIS Yersiniosis occurs in a variety of domestic animals. after liver ii. including ranunculas. clinical findings. • VIBRIOSIS See Campylobacteriosis' VITAMIN DEFICIENCY Diagnosis Nutritional history. pigs) Species Sample Units Deficient Deficient Vitamin A Vitamin E Sheep Serum or µmol/L < 0. all samples for analysis should be protected from light by being wrapped in foil or brown paper and kept chilled during transport to the laboratory. heparin plasma or EDTA plasma Bilirubinuria • hepatocellular disease including infections and toxins • bile duct obstructions • jaundice due to haemolysis. turnips. including babesiosis. pseudotuberculosis or Yersinia enterocolitica. 20 g liver damage occurs. Yersiniosis can be manifest as one or more of the following conditions: VETERINARY LABORATORY MANUAL NB Ensure you obtain at least a full vacutainer of blood before separating off the serum/plasma. including: • ketosis and pregnancy toxaemia in ruminants • starvation in pregnancy. sheep. response to treatment. leptospirosis • chronic copper poisoning • postparturient haemoglobinuria • sulphonamide treatment • plant poisoning. Specimens required As vitamins A and E are light sensitive and degraded in tissues after collection. nephrosis. Disease is usually associated with Yersinia . broom • azoturia (myoglobinuria) • acute nephritis. anorexia • diabetes mellitus • acidosis • prolonged vomiting and diarrhoea • febrile and cachexic diseases • after ether or chloroform anaesthesia Haematuria. privet.9 < 2. renal infarcts. • • • • • • Enterocolitis and diarrhoea Systemic abscesses and mortality Placentitis.6 Cattle Liver µmol/kg < 11.3 plasma Cattle Serum or µmol/L < 0. Specimens will be tested at a specialist laboratory when these tests are requested. 5 ml of serum. specific entities have been associated with Y pestis in domestic carnivores (plague) and Y ruckeri in fish (enteric redmouth).6 plasma Sheep Liver µmol/kg < 700 < 4. If inadequate volumes are presented. However. i. biochemistry.

diarrhoea. mastitis and epididymo-orchitis should be collected as appropriate to the syndrome under investigation. pneumonia. pseudotuberculosis: • Enterocolitis. enterocolitica • Enterocolitis. ill thrift • Systemic myositis (turkeys) Diagnosis Clinical signs. Samples of fixed internal organs with lesions should be taken if systemic disease is suspected. mesenteric lymphadenitis. Portions of tissue fixed in neutral buffered formalin for histopathology. intestinal contents (particularly ileal contents) and fresh tissue submitted chilled for bacteriology. turkeys. histopathology. diarrhoea. Segments from all levels of the intestinal tract. abortion. particularly ileum and caecum. bacteriology. perinatal mortality. mild diarrhoea • Mild systemic abscessation Y. Samples suitable for diagnosis of other causes of diarrhoea. mortality Deer Y. diarrhoea. Y ruckeri is of regulatory importance in fish. Recovery of Y pestis is important in view of the zoonotic potential of this bacterium. abortion. pyaemia. ‘flood mud scours’. ducks. chronic enteritis. pseudotuberculosis • Mild enterocolitis. pseudotuberculosis • Enterocolitis. diarrhoea. iii. VETERINARY LABORATORY MANUAL 104 . perinatal mortality • Epididymo orchitis Y. ii.Cattle Y. enterocolitica • Mild enterocolitis. infertility • Systemic abscessation • Mastitis Y. splenomegaly. mortality • Systemic abscessation. as well as mesenteric lymph nodes should be taken in cases of enterocolitis. mild diarrhoea Poultry (domestic fowl. pseudotuberculosis • Enterocolitis. enterocolitica • Enterocolitis. enterocolitica • Enterocolitis. abortion. ill thrift. enterocolitica • Abortion Sheep Y. diarrhoea. mortality • Chronic systemic abscessation. ‘pyaemic hepatitis’ • Placentitis. Specimens required i. mortality • Placentitis. abortion. Samples of faeces. endometritis. Recovery of Yersinia spp from the alimentary tract and internal organs using routine and selective media. enteritis. mortality • Systemic abscessation Pigs Y. geese) Y. pseudotuberculosis and Y. ill thrift Goats Y. mortality • Placentitis. NB Many Yersinia spp inhabit the intestinal tract of clinically healthy animals and a diagnosis of enteric yersiniosis is confirmed by finding typical histological lesions in sections of the intestine and internal tissues. ill thrift. diarrhoea. perinatal mortality • Pneumonia • Mastitis Y. enterocolitica • Acute septicaemia. diarrhoea.

For the culture of Mycoplasma spp. preferably overnight. COLLECTION OF SPECIMENS Specimens required i. For the investigation of most diseases. iii. v. whole birds or fresh tissues are required. please contact the VETERINARY LABORATORY MANUAL SEROLOGICAL TESTS AVAILABLE FOR POULTRY Serological tests are currently available at EMAI for the following diseases: • Avian encephalomyelitis (AE) • Avian Influenza (AI) • Big Liver and Spleen Disease (BLS) • Egg Drop Syndrome 76 (EDS) (Haemagglutinating avian adenovirus. whole birds. The Specimen Submission Form should be attached to the outside of the outer container and should prominently warn that chlamydiosis or tuberculosis is suspected. For bacterial or fungal culture. For serology. For virus isolation particularly for NDV and AI. Blood should be collected into suitable containers and allowed to clot. Sick birds must be able to withstand transport.swabs in PBSG (which is available in 5-ml plastic screw-top containers from the RVL Menangle /EMAI Virology Laboratory. the field veterinarian should obtain a complete history. the serum should be poured off into a suitable clean container before submission to the laboratory. preferably overnight. The birds must reach the laboratory overnight (summer) or within 24 hours (winter). live birds must be submitted. THE) • Infectious Bursal Disease (IBD) • Marek's Disease (MD) 105 . If they are not likely to survive. The jugular and wing veins are the best vessels from which to collect blood. STANDARD CONTAINERS AND EQUIPMENT See Specimens (General). particular care must be taken to avoid infection of couriers and laboratory staff. ii. and perform several necropsies. with adequate air holes and absorbent lining. Live birds must not be sent. advice before STORAGE AND DESPATCH OF SPECIMENS Live birds should be submitted in pet transport boxes or the like. or were. fixed in formalin and/or fresh. Formalin fixed tissues must be in leak proof containers and packaged securely to prevent breakage in transit. unless arrangements have been made for them to receive water and food in transit. but not frozen. Tissues from necropsied birds. They must reach the laboratory within 24 hours. submit at least 5 affected live or freshly dead birds which are. and must reach the laboratory within 24 to 48 hours. 1 to 5 ml of blood should be collected from each of 10 to 25 birds. The birds must not be overcrowded. Before specimens are submitted. If a diagnosis cannot be made or if laboratory confirmation is required. they should be killed humanely and submitted chilled in an insulated container with an icebrick or ice in a sealed container. iv. Appropriate tests for specific diseases should be requested. examine the flock and affected birds. laboratory for proceeding. In general. or Haemophilus paragallinarum. then appropriate specimens should be submitted with a full history and description of clinical and necropsy findings. If chlamydiosis or tuberculosis is suspected. swabs (with or without transport media) are not suitable. due to loss of viability of fragile organisms and/or overgrowth of contaminants. They must not be frozen. Dead birds should be sealed inside two plastic bags and submitted chilled inside a strong outer container. The blood must not be allowed to haemolyse. fresh or frozen tissues are acceptable. For virology.DISEASES OF POULTRY Many poultry diseases can be diagnosed in the field. are also acceptable. adenovirus 127) • Fowl adenovirus (FAV) • Haemorrhagic enteritis of turkeys (HE. showing typical signs. If unfamiliar with the correct specimens. Fresh tissues and serum should also be transported chilled. If the clotted blood will not reach the laboratory within 48 hours.

serology (pigeons only). for pathology. skeletal muscle. drop). antigen detection in tissues by immunofluorescence (IFAT). for acid fast stain and IFAT. or: ii. for histopathology. or. disease). Specimens required i. as Haemophilus spp quickly lose viability) 106 VETERINARY LABORATORY MANUAL . ii. wet histopathology. for histopathology. Where (i) or (ii) are not feasible. Whole live birds. causing serious illness and sometimes death in humans. gizzard in buffered formalin. Sera. serology. intestinal smears. gross lesions. spleen submitted fresh. for serology.• • • • • Mycoplasma gallisepticum infection (Mg) Mycoplasma synoviae infection (Ms) Newcastle Disease (ND) Reticuloendotheliosis (REV) Salmonella pullorum (Sp) Caution Chlamydiosis is a zoonotic disease. Fresh or frozen sera. histopathology. Specimens required i. AVIAN ENCEPHALOMYELITIS (AE) Diagnosis Young chickens and turkeys Clinical signs (neurological histopathology. paired sera should be submitted to demonstrate rising titres. Whole birds. (Do not submit dead birds. Most serological tests are flock tests. BIG LIVER AND SPLEEN DISEASE Diagnosis Gross lesions. Submit duplicate smears. proventriculus. CORYZA (Haemophilus paragallinarum infection) Diagnosis Clinical signs. CHOLERA See Pasteurellosis' COCCIDIOSIS (AVIAN) Diagnosis Gross lesions. To confirm recent infection. conjunctiva and other representative tissues in buffered formalin. Sera. Liver. Ideally. CHLAMYDIOSIS (AVIAN) Syn: Psittacosis. Portions of affected gut in buffered formalin. examination of tissue impression smears for acid fast organisms. for serology. tissues or swabs for bacteriology. and from conjunctiva. Specimens required i. ii. Unless the birds are vaccinated. collected early during the problem (within 7 days of onset). NB Serology is unreliable for most avian species other than pigeons. iii. iii. or: ii. iii. for microscopic examination for oocysts and schizonts. myocardium. Whole live birds. spleen. isolation. Brain. Specimens required i. The tests are of limited value for individual birds. Whole dead birds. air dried impression smears from the serosal (not cut) surface of liver and spleen. Laying chickens and turkeys Clinical signs (egg production serology. Liver. acid fast and FAT stains. iv. Positive results indicate that the flock has been exposed at some time to the infectious agent concerned. for histopathology. and again during convalescence (3-4 weeks later). the titre range could indicate recent exposure and the paired sera may not be required in all circumstances. Serology is also used in monitoring efficacy of vaccination. either by natural infection (subclinical infection or clinical disease) or due to vaccination. Fresh intestinal scrapings (ie mucosa plus contents). chilled for acid-fast stain and IFAT on fresh smears (made at testing laboratory). ornithosis Specimens required i. bacteriology. serology. for pathology and bacteriology. spinal cord. Whole birds. Diagnosis Gross pathology. iii. Liver and spleen in buffered formalin. histopathology. for histopathology. paired serums are required to demonstrate flock seroconversion or rising flock titres.

or. ii. Whole birds. bursa (if present). heart. for pathology and bacteriology. histopathology. for antigen ELISA and virus isolation. for gross and histopathology. 107 VETERINARY LABORATORY MANUAL . Lesions fixed in buffered formalin. tumours (if present). Fresh trachea. spleen. the range of titres in conjunction with clinical signs would lead to diagnosis. iii. Whole birds. If the flock is not vaccinated. for gross and histopathology. and again during convalescence (3 4 weeks later). Specimens required i. Specimens required i. spleen. Because of relatively poor sensitivity of the antigen ELISA test. Serology to monitor exposure or vaccination efficacy. Bursa of Fabricius in buffered formalin. heart. Specimens required i. histopathology and bacteriology for clinical disease. ii. ovary/testicle. Specimens required i. ovary/testicle. Whole birds. tumours (if present). Whole birds. ii. Trachea. Bacteriology for subclinical disease. virus isolation. kidney. (Do not submit dead birds. sciatic nerves. iii. kidney for virus isolation (and serotyping). Fresh or frozen sera. INFECTIOUS BRONCHITIS (IB) Diagnosis Histopathology.EGG DROP SYNDROME (EDS 76) A drop in egg production due to EDS in layer flocks is usually associated with the appearance of shell-less eggs. virus isolation. histopathology. A drop in egg production without shell-less eggs can be due to a variety of causes. proventriculus. brain. gross pathology. for histopathology. histopathology. or: ii. for histopathology. gizzard. Diagnosis Clinical signs. tissues or swabs for bacteriology. LEUKOSIS Diagnosis Gross pathology. Fresh or frozen tracheas (including larynx) in sterile jars. Many causes can be identified in the field and specimens should only be submitted to confirm the diagnosis of disease caused by infectious agents. for histopathology. ii. Whole birds. Fresh tissues for electron microscopy. Specimens required i. M synoviae infections) Diagnosis Gross pathology. brain. histopathology. MYCOPLASMOSIS (AVIAN) (Mycoplasma gallisepticum. INFECTIOUS BURSAL DISEASE (IBD) Diagnosis Gross pathology. for gross and histopathology. proventriculus. sciatic nerves. kidney. iii. Specimens required i. The following tissues fixed in buffered formalin: liver. for serology. Tracheas (including larynx) in buffered formalin. Specimens required i. FOWL POX Diagnosis Gross pathology. antigen detection in tissues. MAREK'S DISEASE Diagnosis Clinical signs. electron microscopy. as mycoplasmas quickly lose viability). iii. environmental and disease factors. kidney. bursa (if present). lung. or. ii. Specimens required i. pancreas. These include nutritional. collected early during the problem (within 7 days of onset of egg drop). Whole birds. management. INFECTIOUS LARYNGOTRACHEITIS (ILT) Diagnosis Gross pathology. Whole live birds. gizzard. Serology to monitor exposure. without the need for paired sera. lung. histopathology for clinical or subclinical disease. serology. oviduct in buffered formalin. Fresh or frozen sera. The following tissues fixed in buffered formalin: liver. 5-10 tracheas are required.

brain. To check reactors to the field test. heart. isolation of Specimens required i. ii. mycobacteria. NB Do not submit frozen sera. for bacteriology. Sera. Diagnosis Clinical signs. as false positives may occur. Portions of lesions in buffered formalin for histopathology. Specimens required i. heart. kidney. ii. for bacteriology. For an initial flock test. submit fresh sera clearly identified to individual birds. gross pathology. bacteriology for detection of carriers. PULLORUM DISEASE (Salmonella pullorum infection) Diagnosis Serology. or. ii. Preferably off the clot and free of red blood cells. lung and (from ducks) brain). TUBERCULOSIS (AVIAN) Diagnosis Necropsy. bursa (if present).ii. sciatic nerves. for gross pathology and bacteriology. for serology. iii. Fresh tissues (liver. Specimens required i. submit live or freshly humanely killed birds (clearly identified) for bacteriology. P anatipestifer RETICULOENDOTHELIOSIS (RE) Diagnosis Gross pathology. Fresh lesions in sterile sealed jars. To confirm reactors. submit fresh (not frozen) sera from every bird. serology. ovary/testicle. histopathology. ii. iii. proventriculus. The following tissues fixed in buffered formalin: liver. gizzard. tumours (if present). for gross and histopathology. individually identified. spleen. Specimens required i. Whole birds. VETERINARY LABORATORY MANUAL 108 . bacteriology. PASTEURELLOSIS (AVIAN) (Pasteurella infections) multocida. for serology. Fresh sera. Whole birds. histopathology.

do not send live birds.DISEASES OF CAGE BIRDS. after necropsy. COLLECTION OF SPECIMENS Specimens required i. Before submitting specimens. clinical and necropsy findings. ongoing problems involving a number of birds in larger aviaries or pigeon lofts may be investigated. STANDARD CONTAINERS AND EQUIPMENT See Specimens (General). AVIARY BIRDS AND RACING PIGEONS Isolated mortalities or sporadic disease problems in cage and aviary birds and racing pigeons are NOT examined at Regional Veterinary Laboratories. Small birds may. depending on the circumstances. At least two affected live or freshly dead birds which are typical of the flock problem. VETERINARY LABORATORY MANUAL 109 . please phone the RVL to discuss the case. unless a notifiable or exotic disease is suspected. ii. STORAGE AND DESPATCH OF SPECIMENS As for Diseases of Poultry. You should have conducted a thorough investigation of the problem and be able to provide a detailed history. be submitted in toto in 10 times their volume of buffered formalin. If chlamydiosis is suspected. However. fixed in formalin. Tissues from necropsied birds.

If only dead specimens are obtainable. European Foul Brood and many viral diseases affecting both adults and larvae. submit chilled for microbiology. ii. Preferably 30 adult bees submitted live at ambient temperature for microbiology. submitted chilled for microbiology.DISEASES OF BEES Bee diseases are investigated at the Regional Veterinary Laboratories. A brood comb sample containing diseased larvae. Bees should be submitted in aerated containers with a small amount of a candied mixture of sugar and honey. Specimens required ADULT BEE DISEASES i. Diseases seen include American Foul Brood. Four (4) air dried larval smears. LARVAL DISEASES i. Diagnosis Clinical signs and demonstration of causal agent. VETERINARY LABORATORY MANUAL 110 . ii.

• Glass stoppered glass for oxygen. with an estimate of the age or stage of growth. dam.(Phone: 066 240261) Expertise in the diagnosis of fish diseases is currently available at other Regional Veterinary Laboratories. • Sterile 200 ml glass bottles for bacteriology. chilled at 4oC. turbidity. odour and flow. Larger fish are more likely to suffer oxygen deprivation. Record the temperature of the water at several sites and depths. estuarine or ocean environments are important because of their differences in salinity. source or manufacturer of feed. cyanide and algae. Describe the condition of the water in terms of colour. lake. Freshwater. cage or aquarium. Estimates of morbidity and mortality. tank. iii. On fish farms the design and construction of facilities and the source of water should be related to the number and distribution of ponds affected. • Nitric acid washed polyethylene for metals. froth or oil floating at the surface? Does the ecosystem appear "healthy"? iv. A description of the locality of the fish kill should include the type of habitat. Wollongbar can be contacted for advice on the investigation of fish health problems. 38% formaldehyde w/v) Acetic acid DAVIDSON'S SOLUTION Glycerine Formalin (38% formaldehyde) 95% Alcohol 3. and the assistance of officers at these locations can also be sought in the first instance. Submit the results of any onsite water quality tests performed. rate of feeding. poor water quality or exposure to toxins should be suspected. organic debris. located at the Regional Veterinary Laboratory. Fixatives ml BOUIN'S FIXATIVE Saturated aqueous picric acid Formalin (Commercial solution. A general description of the land use in the local drainage basin or watershed may be appropriate. history of dietary changes. mid level and bottom) in the body of water. especially O2 and pH. These measurements. river. could be taken at various sites (inlet. v. STANDARD CONTAINERS AND EQUIPMENT Water sample containers The types of containers recommended for collection of water samples include: • Polyethylene or glass for ammonia. brackish. low or normal? Are algae. pH. Is the water level high. ii. A comprehensive history assists the investigation. pond. • Hexane washed glass for pesticides. middle and outlet) and depths (surface. storage conditions and duration of storage could allow 111 VETERINARY LABORATORY MANUAL . nitrate/nitrite. Details of composition of diet. based on stocking density on farms or the abundance of the species in natural waterways should be provided. Include measurements of length and weight. If other inhabitants of the aquatic environment are affected. stream.5% NaCl solution (or Filtered seawater) Glacial acetic acid Fixation time: 24 hours (minimum) 75 25 5 10 20 30 30 10 HISTORY AND DIAGNOSIS History taking i. and submitted within 24 hours of collection. raceway. whether natural or man made. whereas smaller fish tend to be more susceptible to toxic insults.DISEASES OF FISH The Special Veterinary Officer (Fish Health). filled to exclude air.

The signs exhibited by diseased fish are readily observed in glass sided aquaria. but may be difficult to detect in open water. Feeding patterns may be altered. although these treatments may have eliminated the causative agent. **Microscopic examination of wet preparations of gills. such as infestations with the gill protozoan Chilodonella in many species or epizootic haematopoietic necrosis (EHN) virus in redfin perch. Fish to be submitted are placed in a strong plastic bag inside another plastic bag. together with a half volume of water from the source environment. parasitology. Live moribund or affected fish (preferred specimens). an onsite autopsy should be performed on at least one. Dead fish (rarely of much diagnostic value. the plastic bag should be inflated with oxygen before twisting. STORAGE AND DESPATCH OF SPECIMENS Specimens required i. this increases metabolic rate which increases oxygen consumption and may lead to fouling of the water. On many occasions malachite green or formalin will have been used. List any treatments. Collect fresh and preserved specimens from any autopsies and submit these along with any live affected or freshly dead fish which are available. Any lesions noted at autopsy or during handling should be recorded. fins. The plastic bag is then placed inside an esky and sent to the laboratory. hardness and copper. field assessment of water quality*. flashing or turning upside down may be displayed. iii. clinical signs. ii. eyes. Despatch of several survivors and/or several healthy control fish is often beneficial in reaching a diagnosis. bacteriology. The results of any clinical examination should be detailed. which have been administered to the fish. gills. ii. • Skin colour may Superficial lesions affecting the skin. they may have caused further direct or indirect injury to the fish and these potential complications should be considered. virology. including chemicals and antibiotics. toxicology. Some diseases. What dosage regime (time of treatment. double folding and sealing with strong rubber bands. If only VETERINARY LABORATORY MANUAL 112 . mouth or anus may be visualised in the live fish. because decomposition in fish occurs rapidly and many ectoparasites will detach from the host soon after death. For example. day or season. deaths due to oxygen deprivation are most likely to be clustered in the early hours of the morning when water oxygen levels are depressed. Mortality patterns can be fitted to the time of year. even if chilled. *Water quality assessment is best performed in the field using commercial test kits for ammonia. acidity. histopathology. duration of treatment and concentration or dose rate of chemical) has been applied? Diagnosis History. Diseased fish may change the depth which they normally occupy in the water column or their spatial relation to inlet or outlet points to a pond. unless they are examined on site) Dead fish are virtually useless after transport. Fish should never be fed before transport. as this allows the pathologist to compare affected and unaffected/recovered animals in greater detail. If likely to be in transit for any length of time. Behavioural abnormalities such as gasping at the surface of the water. rendering subsequent examinations negative. unless fish are surfacing. change. Sick fish should also be protected from changes in temperature. COLLECTION. are recurrent seasonal problems. skin and fins should allow common parasites to be identified. vi.nutritional deficiencies or toxicities to be identified. nitrite/nitrate. field evaluation of environmental and ectoparasitic causes of disease**. Diagnostic hints i. If several fish are available for examination.

iii. v. liver. kidney and spleen is often suitable for isolation of viruses. Epizootic Haematopoietic Necrosis (EHN) in redfin perch. In special cases and after consultation specimens can be forwarded to the Australian Fish Health Reference Laboratory. The former is an important pathogen of Salmonids Shellfish Diseases • Discolouration in Abalone Crustacean Diseases • Protozoal myositis Yabbies with ataxia in VETERINARY LABORATORY MANUAL 113 . CRUSTACEANS AND SHELLFISH ALREADY ENCOUNTERED IN NSW Fish Kills Attributed to: • Anoxia • Environmental toxins e. with a section of abdomen removed to allow penetration of fixative to internal organs. Skin Lesions in Fish Often multifactorial. Bouin's fixative (see ‘Fixatives’ above) can be used if the specimens are small. Victoria. • Yellow spot • Red spot in estuarine fish on the North Coast • Goldfish ulcer disease due to Aeromonas salmonicida or Vibrio infections. consider submitting brain. Many examinations for ectoparasites are best carried out in the field using wet preparations. spleen and intestine. bacterial and viral pathogens. SOME DISEASES OF FISH. Rapid blood stains can also be used for smears from fish.dead fish are available for submission to the laboratory. A range of tissues could be taken from larger fish at autopsy. for rapid penetration and optimal preservation. may involve nutritional. iv. sections of the gill arches should be placed in fixative at the start of the examination. Smears of blood and body fluids (for haematology and bacteriology). Frozen fish are unsuitable for histopathology. In addition to internal organs such as heart. As autolysis is rapid in the gills. Fresh tissues (for bacteriological examinations.g. virology and toxicology). concentrating especially on those tissues which exhibit gross pathology. parasitic. even in the presence of formalin. liver. muscle and gonad. Tissues in fixative (submitted in addition to live fish). may be overcome by the use of Davidson's solution (see ‘Fixatives’ above). This ensures that serviceable specimens will be available should there be transportation delays that result in death or deterioration of other specimens. superphosphate • Infectious diseases e. kidney. bacteriology is precluded (culture must be undertaken within 1 hour of death to be of value). Ten percent neutral buffered formalin or 5% formol saline are suitable fixatives for fish tissues.g. submit: • Fixed tissues for histopathology (must be fixed on site) • Frozen tissues for toxicology/virology. A pool of fresh chilled brain. skin. The tendency of some ectoparasites to detach from gills and skin. Swabs are less suitable. Causative agents include: • Bacterial infection with Aeromonas hydrophila in hatcheries. Tissues from large fish (which have been freshly killed) may be removed and transported chilled in sterile containers. Small fish may be immersed in fixative whole. Geelong. Histopathology of lesions has been of value. seen in stressed adult fish. If only dead fish are submitted.

beef and dairy producers. mixed rations. P. The outside of the bag should be labelled with: • Your name • Sample type (hay. pasture. SUBMISSION OF SAMPLES All samples must be accompanied by a Feed Quality Service submission form The form is available from the Feed Quality Customer Service Unit and can be sent to you by fax. silage. The charges are based on which analyses are requested. Charges are made for evaluating feed samples. • Sampling pasture: Sample at random 15-20 locations across the paddock. grain.FEEDS AND PASTURE ANALYSIS NSW Department of Primary Industries provides an independent Feed Quality Service to sheep. SAMPLE COLLECTION A standard procedure for the collection of samples should result in a representative sample and minimal sample deterioration. • Grain and feed mixes: 10-15 grab samples from different locations or bags within the supply. pasture. hay. e-mail or by post. Mix the sample thoroughly and freeze approximately 500g in a plastic bag and send to the laboratory.g. Zn) • pH (silage only) • Ammonia-Nitrogen (silage only) Other specialised tests are available. in accordance with Departmental policy. Ca. fresh crop (green leaf) and fresh pasture must be sealed in a plastic bag (air tight) and frozen immediately. The main points to be considered are: • Samples collected must be representative of the feed and not taken from one bale or bag. Detailed instructions for the collection of samples are available from Livestock Advisory Officers or directly from the Feed Quality Customer Service Unit on 02 6763 1187.) with as much air squeezed out as possible. or freeze the sample and send to the laboratory. browse and leaves. Cu. Each sample should be sealed in a unique. Mix the sample thoroughly and send approximately 500g in a paper bag to the laboratory. NO SAMPLE WILL BE TESTED WITHOUT A CORRECTLY COMPLETED SUBMISSION FORM. The Feed Quality Service accepts most types of feeds for analysis e. • Sampling Hay: 10-15 core or grab samples from at least 10 small or 5 large bales selected at random need to be sampled. Mix thoroughly and send approximately 500g to the laboratory. A limited range of analyses is available for pig and poultry feeds. These samples should then be packed in an esky and sent to the Feed Quality Service. Mix the sample thoroughly and prepare approximately 500g by either: microwave the sample on high for no longer then 30 seconds. contact the Customer Service Unit on 02 6763 1187. Na. DO NOT DESPATCH FROZEN SAMPLES ON THURSDAYS OR FRIDAYS All samples should be sent to: Feed Quality Service NSW Department of Primary Industries RMB 944 Calala Lane Tamworth NSW 2340 114 . The sample must reflect the pasture consumed by the animal. Mn. Fe. strong paper or plastic bag (not freezer bags etc. NB. DO NOT SEND SAMPLES IN GLASS. crop residues. • Sampling Silage: 10-20 grab samples from fresh silage or from at least 10 small or 5 large selected at random need to be sampled. VETERINARY LABORATORY MANUAL TESTS AVAILABLE Individual and package tests are available from the following list: • Dry Matter • Nitrogen (crude protein) • Digestibility • Organic matter • Metabolizable energy • Acid detergent fibre • Neutral detergent fibre • Lignin • Soluble carbohydrate • Fat • Nutrient minerals (B. K. S. For information on analyses and charges. silage etc) • A unique sample number or name Wet samples such as silage. Mg. or dry at 600C for 24hrs.

CHARGES FOR FEED QUALITY SERVICES DO NOT SEND ANY MONEY with the sample. The work load can then be anticipated and a potential turn around time can be given to the officer involved. An invoice will be issued with the laboratory report. e. Discounts for large number of samples may be available.NOTIFICATION TO LABORATORY The Feed Quality Service should be notified by telephone (02) 6763 1187 when samples are sent for analysis. Contact the Customer Service Unit (02 6763 1187) for more information. VETERINARY LABORATORY MANUAL 115 . silage and other fresh material (as noted above) needs to be cooled or frozen.g. Any special conditions needed for sample transport can be discussed at the time of notification.

wells. chloride. NSW Department of Primary Industries' Environmental Laboratories provide a water testing service for farmers and graziers wishing to determine the suitability of their water for agricultural and domestic applications. Testing for nutrients. salinity. iron and salinity. Sampling kits are available from all Departmental offices.au/reader/daslaboratory/das-farm-water.htm Water for human consumption NSW Department of Primary Industries does not undertake testing of water for human consumption. alkalinity. hardness. rivers and farm dams can vary widely. creeks. and can affect irrigation equipment. Common problems include hardness. turbidity.WATER TESTING The quality of water from bores. or the NSW Department of Health's Water Laboratory.gov.nsw. Contact either the Health Inspector at the Local Council. Water quality can impact on plant growth and animal production. pesticide residues and blue-green algae is also available. Further information on water testing is available from the NSW Department of Primary Industries website at http://www. Lidcombe regarding any testing for human consumption: NSW Health Division of Analytical Laboratories Weerona Road LIDCOMBE NSW 2141 VETERINARY LABORATORY MANUAL 116 . saturation index. Standard laboratory reports provide detailed information on pH. sodium absorption ratio and electrical conductivity.agric.

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