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User Guide - South Dakota State University

1. Samples which are frozen or which are 2 113 F on receipt are not acceptable for Salmonella testing E coli 0157 H7 Ground beef Culture using BAX PCR l ti M Th Beef Trim N 60 e Separation Listeria monocytogenes Ready to eat meats BAX PCR Culture M W Antibiotic Residue Kidney Inhibition Swab KIS No formalin fixed tissue will be accepted Kidneys may be frozen prior to shipping but note that testing will be delayed while the tissue thaws Kidney bovine or porcine M F before 2pm Page SUBMISSION GUIDELINES o Samples should be received by the ADRDL no later than the fourth day after sample collection by Meat Inspection personnel o Holiday sample submission procedures will be determined on a case by case basis Visit http www sdstate edu vs foodsafetymicrobiolab index cfm for the latest holiday food safety submission schedule o All samples should be maintained at refrigeration temperatures 2 8 C after collection and until received at ADRDL Insulated shipping containers and ice packs must be used as appropriate FOOD SAFETY INDUSTRY TESTING The following tests are available to personnel from Meat Production facilities and area food producers processors These tests may also be requested by ADRDL pathologists in special situations FOOD SAFETY INDUSTRY TESTING ACCEPTABLE TEST TEST SET UP SAMPLES METHOD DATES Listeria spp Food c
2. Preferably samples of feed from the troughs or bunks should be taken It is necessary to obtain samples of feed from several different locations in the bin crib or silo The single most important step in feed analysis is the sampling procedure Many mycotoxins for example are found in isolated pockets of the feed and therefore could easily be missed if a single sample is obtained PLANTS SUBMITTED FOR IDENTIFICATION Plants submitted for identification should be collected intact with roots and any flowers buds or fruits included Plants or portions of plants are best shipped flat compressed by heavy boards i e inch plywood if possible WATER ANALYSIS WATER One quart of the suspected water collected in a clean glass jar and properly labeled should be submitted Algae poisoning investigation should include the following in the submission a One quart of water in clean container sample should include as much algae as possible keep cool when shipping b 10 cc of algae water in 10 cc of 10 formalin Collect water from thickest part of bloom right off top In general fluids for toxicological analysis should be shipped in new containers when possible Detergent and other residues on glass which has been washed but not rinsed with acid solution and distilled water can confuse analytical results However many times a sample is better than no sample so take available container and rinse it at least three times with liquid to be
3. South Dakota Animal Disease Research and Diagnostic Laboratory Ask for Dr Regg Neiger 605 688 5171 ASPCA Animal Poison Control Center Hotline 888 426 4435 http www aspca org site PageServer pagename pro apcc When you have guestions please call before collecting and sending samples Different types of analyses reguire different specimens and we want to serve your needs in the most efficient manner possible In most cases a complete diagnostic examination is necessary to establish the cause of death Therefore the samples for histopathology should include brain lung heart liver spleen kidney urinary bladder gastrointestinal segments and samples of any organ with gross lesions In addition brain lung liver spleen kidney and any organ with lesions should be submitted refrigerated for culture for bacteria and viruses REMEMBER THE SAMPLES REQUIRED MOST FREQUENTLY FOR TOXICOLOGY ARE LIVER KIDNEY BRAIN STOMACH CONTENTS URINE BLOOD AND SERUM FEED WATER AND ANY OTHER SUSPECTED TOXIC AGENTS SUCH AS PLANTS OR DRUGS IF SUBMITTING SERUM DO NOT USE CLOT SEPERATOR TUBES PULL SERUM OFF BEFORE SHIPPING PLASTIC SNAP TOP TUBES ARE PREFERRED FOR SHIPPING THE SERUM RED RUBBER TOP TUBES MAY AFFECT THE ZINC VALUES SAMPLES GENERALLY REQUIRED What and how much If animals have died fresh and formalin fixed tissues should be collected The tissues generally required for toxicological analysis and the amount of tissues we woul
4. especially for small samples these soak up much as the available serum during separation Page 53 LIPEMIC SERUM Grossly lipemic serum interferes with many serum chemistry tests We employ a detergent separation method to clear lipemic samples this method is routinely performed an all lipemic sera and an additional fee is applied URINE AND FLUIDS Collect urine into a sterile container and chill immediately do not freeze An optimal samples size is 10 mL but to 2 to 3 mL may be adequate for routine testing Fluids collected by aspiration may also be submitted in this manner but if the sample appears to be cloudy viscous or highly cellular also submit at least a portion of the fluid in an EDTA tube to retard clotting and better preserve cells for examination FECES Submit feces in a sealed bag or vial chilled to wet ice temperature do not freeze Open containers or containers grossly contaminated with feces on the outside may not be accepted by the shipping agent If fecal examination for parasitic ova and cysts is requested but delay in shipping is anticipated fix a portion of the fecal sample in parasite fixative see below PARASITOLOGY SAMPLES Collect adult helminths and ectoparasites into a vial and fix in a mixture of 50 alcohol preferably methanol and 50 working strength formalin usually 10 buffered formalin If fixatives are not available ship the sample chilled on wet ice Ectoparasites may also be s
5. may result in false negatives Presumptive positive cultures are now analyzed via PCR at ADRDL for confirmation that T foetus is present Rarely trichomonads other than T foetus are cultured from the prepuce For the InPouch TF system ideal temperature for a loaded culture pouch is 59 98 degrees F Do not ship inoculated culture pouches with ice packs as the organisms are very sensitive to cold temperatures Additionally do not store unused pouches in refrigeration since cold temperatures can actually degrade media ingredients SAMPLING TECHNIQUE FOR TRITRICHOMONAS CULTURE An optimal sample is obtained with an 18 or 21 inch dry infusion pipette attached to a 20 cc syringe A new pipette and syringe are used for each animal to avoid cross contamination The best sample from bulls can be obtained by scraping the distal penis fornix area and prepuce with the end of the pipette Negative pressure is applied with the syringe while scraping to draw loosened material into the pipette Only a small amount of material in the pipette is needed 0 5 to 1 mL Anestrous cows can be sampled using the same device introduced into the anterior vagina and scraping in a similar manner However cervical mucus or vaginal discharge provides a better sample if available Page A satisfactory alternative sample for either cow or bull can also be obtained using a long guarded swab for collection from the same areas The contents of the swab
6. ELISA M thru F Same day Bovine Respiratory Syncytial Virus VN M M Bovine Viral Diarrhea 1 amp 2 VN Th Brucella Card M thru F Plate Same day Rivanol BAPA Same day EHD AGID M thru F 24 hrs Infectious Bovine Rhinotracheitis VN W Johne s ELISA M thru F Same day Leptospirosis 5 serovars MA M Next day Neospora ELISA M thru F Same day Parainfluenza 3 PI 3 VN Th All samples must be received by 10 30 a m or they are not done until the next working day Schedules are subject to variation due to volume of samples received at the lab All classification for Brucellosis on all species must be done by the state s Animal Industry Board Page 35 CAPRINE SEROLOGIC TESTS amp SCHEDULE SEROLOGIC TEST TYPE OF TEST SET UP READ OUT Caprine Arthritis Encephalitis AGID M T W W Th F B abortus Plate On demand Same day All samples must be received by 10 30 a m or they are not done until the next working day EQUINE SEROLOGIC TESTS amp SCHEDULE SEROLOGIC TEST TYPE OF TEST SET UP READ OUT Equine Infectious Anemia AGID M thru Th 24 hrs ELISA M thru F Same day Potomac Horse Fever Forwarded to Protek Western Eastern Venezuelan Encephalitis VN Forwarded to NVSL Ames IA HI Equine Rhinopneumonitis VN Forwarded to CSU Equine Arteritis VN Forwarded to Colorado State Mare Immunopregnancy ELISA Forwarded to CSU Equine Protozoal Myel
7. Graham sdstate edu for questions about submitting tissues and or slides for a specific IHC test or for information about special projects research applications HISTOLOGIC PROCESSING Tissues for research applications can be processed and embedded in paraffin blocks to meet your specifications Please call 605 688 5171 for details and prices SPECIAL HISTOLOGIC STAINS Whenever needed the case coordinator may order special stains in order to better visualize the morphologic characteristics of an etiologic agent usually bacteria or fungi Special stains may also be performed as part of a research project Please call 605 688 5171 for details and prices Page BACTERIOLOGY amp MYCOLOGY GENERAL GUIDELINES Specimens for bacteriology should be collected as soon after the death of the animal as possible and as aseptically as possible They should be placed in heat sterilized containers or sterile plastic bags and sealed tightly Do not use plastic gloves or sleeves Do not chemically treat containers for shipping specimens Sterile tubes for mastitis cultures are available from the laboratory Include a representative portion of the tissues and include any lesions that may be present Place samples of body organs in containers separate from intestine or stomach This prevents contamination by leakage of the intestinal contents It is best to collect specimens of other organs for bacterial culture before opening the gastrointestin
8. PERFRINGENS GENOTYPING E COLI GENOTYPING Page 45 JOHNES DISEASE LAWSONIA INTRACELLULARIS MYCOPLASMA BOVIS MYCOPLASMA HYOPNEUMONIAE PRRSV NORTH AMERICAN EUROPEAN EUROPEAN LIKE PRRSV SCREENING ASSAY QUANTITATIVE ASSAY Page 4 TRITRICHOMONAS FOETUS SEQUENCING SERVICES AVAILABLE Page 46 DNA SEQUENCING CLINICAL PATHOLOGY Page 47 HEMATOLOGY COMPLETE BLOOD COUNT BLOOD TYPING COOMB S TEST URINE FLUIDS CYTOLOGY SERUM CHEMISTRY Page 49 CUSTOM PROFILES Page 50 SPECIAL CHEMISTRY AND IMMUNOLOGY ASSAYS PARASITOLOGY AND FECAL ELISA TESTING Page 52 SAMPLE SUBMISSION Page 53 TEST INTERPRETATION Page 55 LABORATORY NORMAL RANGES FOR CLINICAL CHEMISTRY TRACE MINERALS Page 57 TOXICOLOGY Page 58 GENERAL INFORMATION SAMPLES GENERALLY REQUIRED WHAT AND HOW MUCH WATER ANALYSIS Page 59 SAMPLES REQUIRED FOR SPECIFIC TOXINS TRACE MINERALS Page 62 FEED ANALYSIS FOOD SAFETY MICROBIOLOGY Page 62 MEAT INSPECTION PROGRAM TESTING FOOD SAFETY INDUSTRY TESTING Page 64 APPENDICES Page 65 CURRENT FEE SCHEDULE QUALITY MANUAL SUBMISSION FORMS Page 5 GENERAL INFORMATION SERVICES amp MISSION The mission of the SDSU ADRDL is to provide high quality veterinary diagnostic services as a means to promptly and accurately establish causes of animal health problems Such diagnoses will aid attending veterinarians and health officials in the treatment control prevention and surveillance of animal dis
9. cell culture and identified by EM BOVINE Coronavirus Parvovirus Rotavirus BVD BRSV IBR PI 3 EQUINE Rhinopneumonitis FLUORESCENT ANTIBODY TESTS AVAILABLE PORCINE OVINE PRRS virus Border Disease Swine influenza PI 3 Parvovirus Rotavirus PRV pseudorabies TGE FELINE CANINE Herpesvirus Adenovirus Infectious Peritonitis Coronavirus Panleukopenia Distemper Herpesvirus Parvovirus Other viruses not listed above can be isolated in cell culture and identified by EM Page MOLECULAR DIAGNOSTICS PCR ALL SAMPLES MUST BE RECEIVED BY 11 AM ON SET UP DAY AVIAN INFLUENZA SCREENING Specimen Tracheal or Cloacal Swabs Submit all labeled swabs in leak proof containers and on cold packaging The PCR assay is set up once weekly on Thursday BOVINE LEUKOSIS VIRUS Specimen Whole Blood or Milk Submit all labeled samples in leak proof containers and on cold packaging The PCR assay is set up on the 1 and the 3 Monday of each month Specimen Whole Blood Serum Semen Ear Notch Nasal Swabs Tissue Submit all labeled samples in leak proof containers and on cold packaging Up to 20 whole blood samples can be pooled together for one test sample Up to 10 serum samples can be pooled together for one test sample Up to 50 ear notches can be pooled together for one test sample Submit fresh ear notches in red top tubes Bulk milk tank samples can be tested Do NOT pool bulk tank collections The PCR ass
10. fee schedule Our goal is to provide an initial diagnostic report on surgical biopsies within one to two working day of sample receipt Please contact the lab for more information When shipping biopsy samples please be sure to replace the formalin jar into the whirl pak bag provided and to write any case information on the bag not the jar Jars that have been written on cannot be re used and you will be charged for the jar See http www rottweilerhealth org pdfs april biopsy castle 02 pdf for an example of how to correctly orient punch biopsies ASPIRATES amp IMPRESSIONS Cytologic specimens can be examined in a variety of ways depending on sample characteristics site of collection and potential diagnoses If there is an aspirate fluid at least 0 5 mL the sample should be placed into an appropriate sized EDTA tube and sent to the lab on ice do not freeze These samples will be tested by fluid analysis Smaller samples can be spread onto slides and left unstained then shipped to the lab Sending stained slides to the lab is less ideal since everyone s stain methods are different and reading an atypical stain will make interpretation a problem Unstained slides may be shipped in the protective slide mailer provided with the biopsy shipping box Page FIXATION WITH NEUTRAL BUFFERED FORMALIN 10 formalin is made by adding 9 parts water to 1 part 37 40 formalin 37 to 40 formalin 100 ml Distilled Water 900 ml Sodiu
11. most cases FECAL SEDIMENTATION Sedimentation of the fecal sample to either a accurately count nematode ova to evaluate the effectiveness of control strategies or b detect lungworm larva which may be present in the feces CRYPTOSPORIDIUM TESTING Detection of cryptosporidial oocysts by examination of defined fecal smears stained with a modified acid fast technique ECTOPARASITE EXAM Examination of clinical material for the presence of arthropod parasites Sample can consist of either hair and dander for external parasites or skin scrapings for burrowing and follicular mites PARASITE IDENTIFICATION Whole parasites or environmental pests can be submitted for identification The identification of helminth and arthropod parasites based on morphology of the adult stage often plays an important role in life cycle intervention and other control programs HEMOPARASITE EXAM Examination of the blood smear for evidence of Anaplasma Mycoplasma Ehrlichia Cytauxzoon and other parasitic forms present in the peripheral blood Either unstained blood smears or whole blood in EDTA may be submitted NOTE The familiar blood parasites Haemobartonella and Eperythrozoon have been reclassified as Mycoplasma in the current literature HEARTWORM TESTING Enzyme linked immunodetection of heartworm specific antigen HSA in peripheral blood for the diagnosis of florid or occult heartworm infection in dogs or cats Microfilarial Knott s testing may
12. nights or weekends and left in this cooler for testing the next business day The cooler is adjacent to the loading dock on the east side of the building The on call diagnostician can be reached at 605 690 1576 if problems or questions arise 7 Testing after hours weekends or holidays IS NOT AVAILABLE at the ADRDL See information from the State Public Health Lab Page FEE POLICY DOMESTIC ANIMALS The fee is 40 for South Dakota clients and 46 for out of state clients This fee includes not only the rabies FA test but also routine histopathology and additional laboratory testing such as virology and or bacteriology if requested or found necessary to determine the cause of the animal s death A 17 necropsy fee is added if a necropsy is requested for the purpose of further diagnostics If needed toxicology testing fees are extra WILD ANIMALS Wild animals that originated in South Dakota and have caused a significant risk to human health see definition below will be accepted for rabies testing at NO CHARGE to the submitter The South Dakota Game Fish and Parks Department pays for the testing under these circumstances and only the rabies FA test is completed no additional testing Wild animals that have not caused a risk to human health can be submitted for rabies testing but the submitter will be charged the same fee as for domestic animals If adult bats are submitted with bat pups baby bats only the adults will
13. o Toxin genes heat stable STa Shiga toxin 1 Stx1 and Siga toxin 2 Stx2 e Porcine genotypes of pathogenic E coli isolated from pigs with colibacillosis are listed in order of decreasing frequency of PCR identification o Fimbriae genes K88 F18 F107 or 2134P F41 and K99 o Toxin genes heat labile LT heat stable STa and STb Shiga toxin 2e Stx2e o Both hemolytic and non hemolytic strain may cause neonatal colibacillosis in pigs Post weaning colibacillosis is almost exclusively caused by hemolytic E coli strains JOHNES DISEASE e Specimen Fecal Material please submit at least 5 grams e Submit all labeled samples in leak proof containers and on cold packaging e Please do NOT submit samples in gloves Additional processing charges may apply e The PCR assay is set up on the 1 and the 3 Monday of each month LAWSONIA INTRACELLULARIS e Specimen Porcine Fecal Material or Intestine please submit at least 1 gram e Submit all labeled samples in leak proof containers and on cold packaging e The PCR assay is set up on the 1 and the 3 Monday of each month MYCOPLASMA BOVIS e Specimen Milk Joint fluid tissues or culture a primary culture will be grown at the lab e Submit all samples in leak proof containers and on cold packaging e The PCR assay is set up once weekly on Thursday MYCOPLASMA HYOPNEUMONIAE e Specimen Fresh Lung Tissue or Nasal Swabs using a regular guarded bacti swab e Submit all labeled samples in
14. regarding carcass disposition REPORTABLE DISEASES The Animal Industry Board is required by state law to establish a list of reportable and quarantinable diseases each year on July 1 Reports of such diseases must be made immediately by telephone fax or writing by licensed accredited veterinarians and by all diagnostic laboratories who diagnose diseases on this list Copies of this list shall be mailed to Page 10 all licensed and accredited veterinarians and to diagnostic laboratories on July 1 of each year and are available at no charge from the South Dakota Animal Industry Board 411 South Fort Street Pierre South Dakota 57501 The board may keep such reports confidential except for those reports concerning diseases that are specifically regulated for mandatory control and eradication to protect the public health other livestock or wildlife Source 27 SDR 96 effective April 1 2001 General Authority SDCL 40 3 14 40 5 7 40 5 8 6 Law Implemented SDCL 40 5 DIAGNOSTIC TESTING STANDARD DIAGNOSTIC EVALUATION m Specimens are processed on the day they are received with very few exceptions The time frame for processing specimens and receiving results in the laboratory is as follows Tissues sent by mail UPS or bus are usually received in 1 2 days FAT results are completed on the day of receipt or the following morning Bacterial isolation requires 1 2 days excluding contamination o Sensitivity testing takes another da
15. 21 11 17 8 12 17 5 MCHC 32 36 30 36 34 36 30 34 30 0 36 8 30 0 39 5 Platelets 10 mm 200 500 300 800 130 300 130 500 100 800 270 750 Fibrinogen mg dL 200 400 50 300 100 400 100 500 300 700 100 500 Pl Protein g dL 5 5 8 0 60 75 5 8 87 60 80 70 85 6 0 7 5 Page PARASITE BURDEN SCORING FOR FECAL FLOTATION AND CRYPTOSPORIDIA EXAMS scores assigned by ADRDL based on flotation of 1 g of feces or standard Cryptosporidia smear SEMI QUANTITATIVE COCCIDIA CRYPTO TRICHURUS amp OTHER SCORE OOCYSTS SPORIDIA NEMATODIRUS NEMATODE OOCYSTS OVA OVA Rare 1 1 to 2 HPF 1to2 slide 1t03 slide 1 to 10 slide Light 2 2to10 2 to 10 slide 4 to 6 slide 11to50 HPF slide Moderate 3 10 to 20 10 to 20 7 to 10 slide 51 to 150 HPF slide slide Heavy 4 gt 20 HPF gt 20 slide gt 10 slide gt 150 slide TRACE MINERALS TRACE MINERALS A minimum of 4 ml of serum is needed for a complete mineral screen Blood needs to be spun down and the serum drawn off before the sample is shipped or frozen Plastic snap top tubes are preferred for shipping the serum Red rubber top tubes may affect the zinc values Do not use clot separator tubes Freezing of whole blood ruptures the cells and causes hemolysis Hemolyzed serum yields questionable results selenium and iron values will be falsely elevated and other minerals may also be affected Page TOXICOLOGY GENERAL INFORMATION REFERENCE CONTACTS
16. 8 4 5 3 5 5 0 3 8 5 8 3 9 5 4 CL mmol L 96 119 100 120 90 102 91 106 86 109 95 103 CA mg dL 9 12 8 5 11 3 10 6 13 0 9 12 8 0 10 5 11 5 12 8 PO4 mg dL 2 9 6 4 3 6 8 5 2 7 5 0 5 3 9 6 5 7 9 3 5 7 3 MG meg L 14 2 0 2 7 3 7 1 6 2 3 2 2 2 8 CO2 mmol L 17 24 17 24 24 30 AG mmol L 15 25 15 25 10 20 NOTE Shaded test values are included in the diagnostic profile for that species Values for adult ruminating animals generally greater than 6 months of age See INDIVIDUAL TESTS AND ABBREVIATIONS above May be performed based on other test results see SERUM CHEMISTRY section above Page LABORATORY NORMAL RANGES FOR HEMATOLOGY based on normative data for each species collected and performed by ADRDL or from applicable published data PARAMETER UNITS CANINE FELINE EQUINE PORCINE BOVINE OVINE WBC Count 103 mm 6 17 55 195 5 125 11 22 4 12 4 13 Neutrophils 60 77 35 75 22 72 28 47 15 45 10 50 Bands 0 3 0 3 0 2 0 4 0 2 0 2 Lymphocytes 12 30 20 55 17 68 39 62 45 75 40 75 Monocytes 3 10 1 4 0 7 2 10 2 7 0 6 Eosinophils 2 10 2 12 0 10 1 11 0 2 0 15 Basophils 0 3 0 1 0 4 0 2 0 2 0 13 RBC Count 10 mm 5 5 8 5 50 99 7 11 5 0 9 9 5 15 6 15 Hematocrit 37 55 30 45 34 45 32 50 24 46 27 45 Hemoglobin g dL 12 18 10 15 11 5 16 10 16 8 15 9 16 MCV um3 60 77 39 55 36 49 50 68 40 60 25 42 MCH pg 19 5 30 0 12 5 12 7 17 5 17
17. AL TESTS AND ABBREVIATIONS Chemistries Enzymes Electrolytes Total Protein Alkaline Phosphatase AP Sodium NA TP Albumin Alanine Transaminase ALT Potassium K ALB Globulin Aspartate Transaminase Chloride CL GLOB AST Glucose Gamma Calcium CA GLU Glutamyltransferase GGT Urea BUN Amylase AMYL Phosphate PO4 Creatinine Lipase LIP Magnesium MG CRE Uric Acid Lactate Dehydrogenase Bicarbonate URI LDH CO2 Total Bilirubin Creatine Kinase CK Anion Gap AG TBIL Direct Bilirubin DBIL Total Iron FE Cholesterol CHOL FOOTNOTES FOR TESTS AND PROFILES Calculated from other measured parameters Direct bilirubin is performed as an additional test no charge if total bilirubin is elevated ipase is performed as an additional test no charge if amylase is elevated in the absence of azotemia STANDARD PROFILES For each profile test abbreviations are listed for those tests included Please contact the lab for more information RUMINANT DIAGNOSTIC PROFILE TP ALB GLOB GLU BUN CRE TBIL DBIL AST GGT CK NA K CL CA P04 MG DAIRY MANAGEMENT PROFILE TP CA PO4 MG NEFA BHA Page PORCINE DIAGNOSTIC PROFILE TP ALB GLOB GLU BUN CRE TBIL DBIL FE AST LDH NA K CL CA P04 MG EQUINE DIAGNOSTIC PROFILE TP ALB GLOB GLU BUN CRE TBIL DBIL AP AST GGT CK NA K CL CA PO4 MG LA
18. E HEAD If removal of brain is not possible submission of entire head is acceptable chill before shipment if possible SPINAL CORD Entire carcass vertebral column or sections of spinal cord fresh chilled Sections of spinal cord 4 5 pieces formalin fixed SPINAL FLUID CSF can be collected prior to removal of the skull submit in red top tube chilled OPTIONAL SPECIMENS FOR SUBMISSION WHOLE BLOOD and or SERUM Fresh chilled Lead toxicosis EDTA Organophosphate toxicosis EDTA Calcium Magnesium Copper deficiencies Liver and or kidney disease STOMACH RUMEN CONTENT Fresh chilled Organophosphate toxicosis Lead toxicosis Urea toxicosis freeze content if urea toxicosis is suspected TONSIL Fresh chilled amp formalin fixed Pseudorabies PRRS virus lung and lymph node also fresh amp formalin fixed LIVER Fresh chilled Lead toxicosis Selenium toxicosis Copper deficiency KIDNEY Fresh chilled Lead toxicosis SMALL INTESTINE Fresh chilled amp formalin fixed Edema disease E coli Enterotoxaemia C perfringens type D COLON and or FECES Fresh chilled amp formalin fixed colon only Nervous coccidiosis COMMENTS e If rabies is suspected see RABIES e If submission is for CWD testing see CHRONIC WASTING DISEASE e Cerebellum and brain stem should always be included in submissions of CNS disease Page 16 DO NOT FREEZE fresh brain or head Package fre
19. Heparin tubes from live animals can be analyzed for lead Feed and any suspected materials would also be helpful MYCOTOXIN The samples needed for investigation for mycotoxicosis are feed and or suspected source Many mycotoxins are found in isolated pockets of the feed and could therefore easily be missed if a single grab sample is obtained Collect at least 3 pounds of representative feed for an adequate sample NITRATE AND NITRITE Specimens to submit for nitrate and nitrite analysis are feed forage water plasma and entire eyeball Eyeball should be submitted frozen Rumen contents are of limited value but may be looked at if there are no other options ORGANOCHLORINE INSECTICIDES Organochlorine insecticides such as dieldrin DDT and toxaphene affect the central nervous system Stomach content feed or suspected source brain liver and kidney are the best samples for analysis ORGANOPHOSPHATE INSECTICIDES Stomach contents liver feed and any other suspected source of toxicant are the best samples for the detection of organophosphate insecticides Included in this group of chemicals are phorate Thimet and fonofos Dyfonate PRUSSIC ACID CYANIDE Prussic acid cyanide poisoning is associated with sorghums Sudan grasses milo cane and flax Prussic acid does not occur in corn or millet Except in very immature plants prussic acid accumulates largely in the leaves It is best to submit samples that are not chopped C
20. LE SPECIFIC SYNDROMES Page 12 ARTHRITIS Page 12 AVIAN INFLUENZA Page 13 CHRONIC WASTING DISEASE Page 14 ENTERITIS Page 15 NEUROLOGIC DISORDERS EXCLUDING RABIES Page 16 PNEUMONIA RESPIRATORY DISEASE Page 18 RABIES Page 19 HOW TO SUBMIT RABIES SUSPECT CASES TO ADRDL FEE POLICY RESULTS AND REPORTING SUBMITTING RABIES CASES TO SOUTH DAKOTA PUBLIC HEALTH LAB Page 21 SCRAPIE Page 22 TRICHOMONIASIS Page 23 PATHOLOGY Page 25 BIOPSY SERVICE Page 26 ROUTINE HISTOPATHOLOGY Page 27 IMMUNOHISTOCHEMISTRY Page 28 BVD EAR NOTCH TEST OTHER TESTS AVAILABLE BACTERIOLOGY amp MYCOLOGY Page 30 GENERAL GUIDELINES ANTIBIOTIC SENSITIVITY TESTING SPECIFIC SYNDROMES Page 31 SEROLOGY Page 32 GENERAL SAMPLING PROCEDURES BOVINE SEROLOGIC TESTS Page 34 SCHEDULE FOR BOVINE SEROLOGIC TESTS Page 35 CAPRINE SEROLOGIC TESTS amp SCHEDULE Page 36 EQUINE SEROLOGIC TESTS amp SCHEDULE OVINE SEROLOGIC TESTS Page 37 SCHEDULE FOR OVINE SEROLOGIC TESTS PORCINE SEROLOGIC TESTS Page 38 SCHEDULE FOR PORCINE SEROLOGIC TESTS SMALL ANIMAL SEROLOGIC TESTS Page 39 SCHEDULE FOR SMALL ANIMAL SEROLOGIC TESTS ABBREVIATIONS INTERPRETATION OF SEROLOGIC TEST RESULTS Page 40 VIROLOGY Page 42 GENERAL GUIDELINES SAMPLING amp SUBMISSION GUIDELINES AGENTS DETECTABLE BY ISOLATION Page 43 FLUORESCENT ANTIBODY TESTS AVAILABLE MOLECULAR DIAGNOSTICS PCR Page 44 AVIAN INFLUENZA SCREENING BOVINE LEUKOSIS VIRUS BVD CIRCOVIRUS PORCINE TYPE 2 CLOSTRIDIUM
21. N SEROLOGIC TESTS Numerous different serologic tests are used and in some cases more than one type of test is used for a single antigen These tests vary widely in their capabilities and the titer to an antigen in a serum may be much higher by one test than it is by another For example the agglutination test for H somni shows more sera with titers than the complement fixation test and the titers are consistently higher by agglutination than by CF Killed leptospira antigens show much lower titers than do the live cultures used in the microagglutination test All serologic tests have deficiencies of one type or another It is hardly reasonable to expect practitioners to be aware of all these factors but they should at least know that serologic tests are not infallible and that the results often depend on the kind of test Page 40 used THE VACCINATION STATUS OF THE ANIMAL The antibody titer that results from vaccination varies with the antigen the adjuvant the recipient species previous and subsequent exposure to the antigen and the individual s immunologic reaction to the antigen Because these and other factors affect the titer and its persistence it is extremely difficult to interpret the results of serologic tests on vaccinated animals In addition to the difficulties this presents in using antibody titers in clinical diagnosis antibody titers are not always directly related to resistance to infection Some animals that hav
22. RGE ANIMAL PRESURGICAL PROFILE TP BUN GGT CK SMALL ANIMAL DIAGNOSTIC PROFILE TP ALB GLOB GLU BUN CRE TBIL DBIL AP ALT AMYL LIP LDH NA K CL CA PO4 MG SMALL ANIMAL PRESURGICAL PROFILE ALB GLU BUN ALT AVIAN DIAGNOSTIC PROFILE TP ALB GLU CHOL CA PO4 RENAL PROFILE BUN CRE K PO4 NSAIDS PROFILE BUN CRE AP ALT ELECTROLYTE PROFILE NA K CL CO2 AG CUSTOM PROFILES By indicating your choice of individual tests on the submission form a custom profile can be performed to fit the needs of the situation In general cost discounts are applied to multiple tests requested for a single sample Substitution of individual tests in the profiles listed above is also possible please contact the lab for more information SPECIAL CHEMISTRY AND IMMUNOLOGY ASSAYS THYROID HORMONE T4 Determination of total T4 hormone by radioimmunoassay RIA Currently available for canine feline and equine samples only except as experimental assays please contact the lab for more information CORTISOL CORT Determination of cortisol by radioimmunoassay RIA often as the endpoint analysis for dexamethasone suppression tests Currently available for canine samples only except as experimental assays please contact the lab for more information Page 50 C REACTIVE PROTEIN CRP A marker of hepatic acute phase response of inflammation and infection particularity in pigs Currently available for exp
23. South Dakota State University Animal Disease Research amp Diagnostic Laboratory User s Guid Lo SDSU ADRDL Department of Veterinary Science Box 2175 North Campus Drive South Dakota State University Brookings South Dakota 57007 1396 The materials in this handbook have been prepared by faculty and staff of the Animal Disease Research and Diagnostic Laboratory South Dakota State University Inquiries regarding the handbook and its content should be made to David H Zeman DVM PhD Head Veterinary amp Biomedical Sciences Dept amp Director ADRDL amp OBL OR Tanya D Graham DVM DACVP Associate Director ADRDL c o South Dakota State University North Campus Drive Box 2175 Brookings SD 57007 1396 Phone 605 688 5171 Fax 605 688 6003 TABLE OF CONTENTS CLICK ON THE LINKS BELOW TO GO TO THAT PAGE EXCISIONAL BIOPSIES ASPIRATES amp IMPRESSIONS FIXATION WITH NEUTRAL BUFFERED FORMALIN GENERAL INFORMATION Page 6 SERVICES amp MISSION QUALITY PROGRAM amp CUSTOMER SERVICE COMPUTER BASED CASE REPORTING VADDS Page 7 CONTACT INFORMATION Page 8 MAILING SHIPPING ADDRESS FACULTY CONTACTS WEB PAGE LOCATION amp SUBMISSION INFORMATION Page 9 LABORATORY POLICIES Page 10 RECEIVING amp BUSINESS HOURS SUBMISSION FORMS VETERINARY REFERAL amp REPORTING LIVE ANIMAL SUBMISSION CARCASS DISPOSITION REPORTABLE DISEASES DIAGNOSTIC TESTING Page 11 STANDARD DIAGNOSTIC EVALUATION TEST SET UP SCHEDU
24. ae ELISA M thru F Same day Parvovirus HA W Th Porcine Reproductive and Respiratory IFA only if reguested On demand Same day Syndrome PRRS ELISA test of choice Pseudorabies Screen G1 ELISA M thru F Same day Swine influenza SIV HI M thru Th 24 hrs H3N2 8 H1N1 Transmissible Gastroenteritis VN Detect infection Sent to ISU Virus All samples must be received by 10 30 a m or they are not done until the next working day Page 38 SMALL ANIMAL SEROLOGIC TESTS AGENT TEST PURPOSE INTERPRETATION Brucella canis RSAT Detect infection Positive Agglutination positive sample is sent to Cornell for confirmation Negative No agglutination A positive RSAT and negative 2M 2M RSAT RSAT should be retested in 30 days Feline Infectious Peritonitis Virus FIP ELISA Feline Leukemia ELISA Detect Positive Color development Virus infection Negative No color or carriers Feline T lymphotrophic Virus ELISA Lyme disease IFA Detect Sent to KSU infection SCHEDULE FOR SMALL ANIMAL SEROLOGIC TESTS SEROLOGIC TEST TYPE OF TEST SET UP READ OUT Brucella canis Card On demand Same day Feline Infectious Peritonitis Virus FIP ELISA On demand Same day Feline Leukemia Virus ELISA On demand Same day Feline T lymphotropic Virus ELISA On demand Same day Lyme Disease IFA ABBREVIATIONS A Agglutination AGID Agar gel immunodiffusion test BAPA Buffered acidified p
25. al tract For culturing submit a 4 6 section and don t open the intestine Fluids from body cavities heart sac joints etc are best collected with a sterile swab with transport media and submitted in a sealed sterile tube or aspirated into a sterile disposable syringe Except for slaughter specimens to be cultured for Brucella sp freezing is undesirable Adequate refrigeration should accompany specimens so they reach the laboratory still chilled When in doubt as to what should be submitted the following organs are desirable spleen 1 2 heart unopened jejunum and ileum 3 4 inches securely tied brain longitudinal with brain stem lung including lesion kidney in capsule colon 3 4 inches securely tied lymph nodes adjacent to lesions Liver amp Gall bladder if looking for Salmonella 10 any other tissue with lesions WONAMRWNE Note liver is a poor tissue to submit for culture because it is rapidly invaded by intestinal bacteria after death Clearly label each specimen with the owner s and the veterinarian s name ANTIBIOTIC SENSITIVITY TESTING Specimens from recently treated animals may not yield satisfactory results because of the suppression of the organism by high tissue levels of antibiotic residue Page 30 SPECIFIC SYNDROMES ABSCESSES DRAINING TRACTS It is important that an adequate sample is obtained with a swab Organisms collected on swabs are few compared to those in tiss
26. also be done by special request Page 52 TRITRICHOMONAS TESTING This examination is performed mainly on preputial wash material collected into a special transport nutrient media following 6 days of incubation Please see the Trichomonasis Testing section of this guide for more information FECAL ELISA ASSAYS Although not traditional parasitology assays fecal ELISA procedures are performed by the Clinical Pathology section as an adjunct to fecal exams and other diagnostic cases All such assays at ADRDL are configured to detect the antigen of interest in fecal samples and do not detect fecal antibody levels Assays include bovine rotavirus A bovine coronavirus giardiasis and canine parvovirus FECAL EXAMINATION Single or pooled fecal samples can be submitted alone for diagnostic workup of enteritis either in a herd health or individual case setting Testing of the fecal sample is at the discretion of the pathologist and depends upon species age and differential diagnoses but may include parasitology exams fecal ELISA testing direct examination of fecal smears aerobic culture anaerobic or microaerophilic culture and or electron microscopy Please see section II D 1 of this guide for more information ELECTRON MICROSCOPY These procedures allow ultrahigh magnification of biological samples for examination purposes The main diagnostic application for electron microscopy is negative staining of fecal samples to detect
27. and describe enteric viruses for which immunoassays are not available The ADRDL also has capabilities to perform research applications such as ultrastructural pathology and scanning electron microscopy please contact the lab for more information SAMPLE SUBMISSION NOTE All samples must be clearly labeled with some individual identification animal name tag number or other and must be accompanied by a submission form to avoid errors in interpretation and reporting WHOLE BLOOD For hematology or other testing requiring whole blood submit 2 to 5 mL of unclotted blood Our methods are optimized for the use of EDTA as an anticoagulant lavender topped tubes contact the lab if there are any questions Ship whole blood on wet ice or ice packs but do not allow to freeze PREPARED SMEARS Blood feces fluids or cytology samples may be submitted as air dried smears Please include at least 2 duplicate smears packed to avoid breakage during shipment Additional unstained smears are useful if multiple staining methods prove necessary SERUM For standard diagnostic profiles draw at least 6 mL of whole blood into a red topped tube and allow blood to clot Once serum is separated by centrifugation chill or freeze serum and ship Each serum chemistry assay requires at least 0 2 mL of serum If the volume of sample is inadequate for all requested tests the lab will contact you to prioritize testing before proceeding Avoid gel separation tubes
28. ay is set up once weekly on Monday CIRCOVIRUS PORCINE TYPE 2 Specimen Serum Lung Tonsil Spleen amp Lymph Node Submit all labeled samples in leak proof containers and on cold packaging The PCR assay is set up once weekly on Thursday QUANTITATIVE PORCINE CIRCOVIRUS PCV2 Specimen Serum Lung Tonsil Spleen amp Lymph node Submit all labeled samples in leak proof containers and on ice packaging For quantitation of tissues gram weight of tissue submitted is needed e g tissue should be submitted rather than homogenate where weight is unknown The PCR assay is set up once weekly on Thursday Positive PCR results will be reported in copies ml for fluid samples and copies gram for tissue samples CLOSTRIDIUM PERFRINGENS GENOTYPING Specimen Feces Intestine or Culture a primary culture will be grown at the lab Submit all labeled samples in leak proof containers and on cold packaging Six toxin types can be detected and the results used to categorize the sample as one of 5 genotypes The PCR assay is set up on the 1 and the 3 Monday of each month Page 44 E COLI GENOTYPING e Specimens Blood agar plate or an agar slant of E coli isolated from the ileum e Expected turnaround time is 7 10 days e Bovine genotypes of pathogenic E coli isolated from calves with colibacillosis are listed in order of decreasing frequency of PCR identification o Fimbriae and other adhesion genes K99 F41 eaeA intimin
29. be tested HUMAN HEALTH RISK DEFINITION The exposure of a human or domestic animal to saliva from the suspect animal either through a bite exposure of mucous Membranes exposure of an open wound or scratches OR The exposure of a human or domestic animal to central nervous system tissue from the suspect animal either through exposure of mucous membranes or exposure of an open wound RESULTS AND REPORTING 1 Laboratory results are reported by telephone as soon as they are available to the referring veterinary clinic listed on the submission form 2 Test results are reported as no test when ANY part of the brain required for testing per CDC guidelines is missing for any reason including autolysis trauma and or only half of brain submitted fresh and the FA result is negative 3 Test results are also reported as no test when brain tissue cannot be identified for any reason most often due to marked autolysis and or severe brain trauma and the FA test is not performed 4 In addition to the referring veterinary clinic all POSITIVE rabies FA results from domesticated animals will also be reported to the State Health Department and Animal Industry Board in the state where the animal resided 5 All POSITIVE rabies FA results from wild non domesticated animals will be reported to the State Health Department the Animal Industry Board and Game Fish and Parks Department in the state where the animal resided additionally the re
30. ccinated animal Bovine Viral Diarrhea VN Detect Positive 4 fold rise in antibody titer between paired sera Virus or Border Disease infection Antibody titer gt 1 4096 usually significant unless vaccinated Brucella Card Detect Positive Agglutination Plate infection Negative No agglutination Tube Rivanol EHD AGID Infectious bovine VN Detect Positive 4 fold rise in antibody infection titer between paired rhinotracheitis virus sera Generally titer gt 1 128 indicates recent infection vaccine titers usually lower Johne s Para ELISA SDSU is currently using the Biocor ELISA test These will be reported tuberculosis as positive or negative with a corresponding OD value Leptospirosis 6 MA Determine Sera screened at 1 100 tittered if agglutination detected Titers serovars titers gt 1 800 may be significant depending on vaccination history Hardjo is the exception and not very antigenic for cattle Neospora ELISA Detect Cannot determine recent or old infection just exposure exposure Parainfluenza 3 virus VN Detect Positive 4 fold rise in antibody titer between paired sera exposure Page 34 SCHEDULE FOR BOVINE SEROLOGIC TESTS TYPE OF SEROLOGIC TEST TYPE OF TEST SET UP READ OUT Anaplasmosis CF M thru F Same day Bluetongue AGID On demand export only 24 hrs ELISA Same day Bovine Leukemia Virus AGID On demand for export only or 72 hrs confirmation of ELISA
31. cks to maintain refrigeration until the specimens arrive at the laboratory NOTE Mycoplasmas and Ureaplasmas do not survive on cotton swabs Dacron swabs with special transport media are required Contact the laboratory if help is needed Diagnosis of CAE in adult goats is by clinical sign joint capsule lesions macro and microscopic elimination of other causes and positive AGID serology results evidence of exposure DETECTABLE AGENTS Bacteria common aerobic amp anaerobic bacteria Chlamydia and Erysipelothrix Mycoplasma Viruses Page AVIAN INFLUENZA SEROLOGY AGID Identifies Type A Influenza Virus Negative AGID Positive Matrix Type A Send serum to NVSL for typing and or No further testing is reguired submit Sule for PCR Sensitivity of the AGID test Not published Turn around time TAT for serology testing is generally 2 to 3 business days PCR SWAB TEST required for ducks and geese Oropharyngeal amp or Cloacal Swabs Negative Matrix Type A Positive Matrix Type A No further testing is reguired ADRDL runs H5 8 H7 PCR tests if HS or H7 PCR is positive Sample s are sent to NVSL for confirmation and to ID the type of Al virus Page CHRONIC WASTING DISEASE SPECIMENS TO SUBMIT Fresh unfixed retropharyngeal lymph node For details about how to collect the retropharyngeal lymph node see http agbiopubs sdstate edu articles ExEx11012 pdf Formalin fixe
32. d like to receive are described below FIXED TISSUE A thick liver section and cross section of kidney should be submitted in 10 buffered formalin for histopathology studies Also the brain should be divided longitudinally and placed in 10 buffered formalin IF YOU NEED RABIES EXAMINATION SUBMIT THE WHOLE BRAIN FRESH DO NOT FREEZE IT Formalin fixed sections of other organs should be submitted as each case dictates If in doubt it is better to submit too many tissues than not enough DO NOT FREEZE FORMALIN FIXED TISSUES Page 58 FRESH TISSUE As a minimum the following fresh specimens should be submitted for analysis Liver A large handful of liver at least 100 grams Kidney One whole kidney or both kidneys in very small animals Brain of the brain unless whole fresh brain required for Rabies Examination Stomach or Rumen Content A double handful about one pound of stomach or rumen content Urine All available up to 200 milliliters ml should be collected in a clean glass jar test tube or syringe Whole Blood Ten to twenty ml of heparinized green top tubes blood and two to twenty ml of serum separated from the clot before shipping are adequate for analysis If heparin tubes not available ETDA purple top tubes may be adequate FEED AND PLANTS INTENDED FOR CHEMICAL ANALYSIS At least one pound of each of the various components in the total ration should be collected and submitted in airtight sealed plastic bags
33. d obex A pictorial obex removal is available at http www aphis usda gov vs nvsl BSE procedure manual pdf The following instructions on obex removal using the scoop method is from Bio Rad s Brain Stem Sampling Procedure in the Slaughterhouse With the head placed flat on the table resting on the forehead and nose The spoon can be easily inserted thru the foramen magnum underneath the dura mater with the convexity of the spoon facing downwards By advancing anteriorly in the axis of the medulla oblongata the anterior lip of the spoon sections the base of the cerebellum isolating the cerebellar peduncles anterior middle and posterior The roots of the cranial nerves are sectioned by rotating the spoon from left to right and from right to left If you have trouble severing the cranial nerves hold the dura covering the brain stem with forceps and slide curved scissors into the foramen magnum to cut the cranial nerves Once you are comfortable using the spoon technique you may not need to use the scissors to section the cranial nerves Page ENTERITIS SPECIMENS TO SUBMIT ALL SPECIES PER ANIMAL Duodenum e One 6 inch length fresh e One 2 inch length fixed in 10 buffered formalin Jejunum amp Ileum e Two 6 inch lengths each fresh e Two 2 inch lengths each fixed in 10 buffered formalin Cecum amp Spiral Colon e Two 2 inch squares each or several partial loops piglet fresh e Two 2 inch squares each fixed i
34. e Progressive Pneumonia AGID M thru Th Toxoplasmosis Agglut T F All samples must be received by 10 30 a m or they are not done until the next working day Page 37 PORCINE SEROLOGIC TESTS AGENT TEST PURPOSE INTERPRETATION Actinobacillus Samples are sent to lowa State University or Biovet for testing pleuropneumoniae Brucella Card Rivanol Leptospirosis MA Determine Sera screened at 1 100 tittered if agglutination detected Titers 6 serovars titers gt 1 800 may be significant depending on vaccination history Mycoplasma ELISA hyopneumoniae Porcine parvovirus HI Detect Positive 4 fold rise in antibody titer between paired sera Ubiquitous infection virus Generally vaccine titers lower field virus high titers gt 1 1 280 Porcine Reproductive IFA Detect Positive 1 20 indicates exposure and Respiratory infection Negative lt 1 20 Syndrome PRRS ELISA Positive or Negative S P value of 0 4 or greater considered positive Pseudorabies ELISA Detect Positive infection Negative Samples screened on ELISA positive samples verified by G1 ELISA test G1 ELISA Swine influenza HI Detect Positive 1 80 and greater SIV infection Suspect 1 10 1 40 Negative lt 1 10 SCHEDULE FOR PORCINE SEROLOGIC TESTS AGENT TYPE OF TEST SET UP READ OUT Brucella Card M thru F Same day Rivanol Leptospirosis 6 serovars MA M Same day Mycoplasma hyopneumoni
35. e no detectable antibody to an infectious agent may still be resistant to infection by that agent On the other hand animals may have antibody to an infection and remain susceptible to that infection THE TITER IN A SINGLE SERUM SAMPLE is only rarely high enough to be diagnostic Most titers will fall into the uninterpretable middle range Serologic testing of the herd vs the individual is much more likely to provide diagnostic information To use serologic examinations most effectively for diagnosis take serum from at least 10 animals or 10 of the herd whichever is greater and 10 21 days later take samples from these same animals If the infection is active in the herd some of the animals that had no detectable antibodies in the first serum should have antibodies in the second sample And some animals that had low titers in the first serum sample should have at least 4 fold titer increases in the second sample Titer changes less than 4 fold are within the normal accuracy of most serologic tests i e a titer of 1 8 could be read as 1 4 in one test and 1 16 in a second test PAIRED SERA from an aborting animal usually will not provide a diagnosis because the peak of the titer has already been reached at the time of abortion Page VIROLOGY GENERAL GUIDELINES 1 Specimens should be collected from sacrificed animals or as soon after death as possible Viruses start losing viability when the animal dies Collect appropriate specim
36. eases to the benefit of the livestock industry other animal owners and society at large We also continually look for research opportunities utilizing the case materials The ADRDL is an accredited AAVLD laboratory and a member of the USDA National Animal Health Network Completing and submitting any submission form or any other means of requesting services creates a contractual agreement for services requested and the specimens submitted become the property of the ADRDL In addition at no additional expense to our clients specimens submitted to the ADRDL may be subjected to additional testing upon the order of state or federal animal health officials or whenever a Foreign Animal Disease is suspected or in support of surveillance for other animal diseases We take pleasure in serving our clients and improving animal health We look forward to your submissions QUALITY PROGRAM amp CUSTOMER SERVICE ADRDL maintains a Quality System compliant to the American Association of Veterinary Laboratory Diagnosticians AAVLD standards The AAVLD accreditation program is based on the OIE guidelines The accreditation process helps us demonstrate and prove our abilities to provide quality diagnostic services to the animal owners of South Dakota and surrounding areas Our quality system has a simple rule SAY WHAT YOU DO DO WHAT YOU SAY PROVE IT Through this rule we strive to make sure that the quality system policies and procedures establ
37. en completed 1 Package the brain in a sterile plastic bag placed inside a crush proof container Submit to the lab in an appropriate leak proof insulated shipping container with adequate ice packs to keep specimen chilled during shipping DO NOT FREEZE the fresh brain 2 As always the laboratory WILL NOT ACCEPT LIVE ANIMALS for rabies testing Whole bodies complete heads or removed brains are all acceptable specimens for submission ADRDL staff will remove brains upon arrival at no additional charge 3 Fill out the standard ADRDL submission form with complete information including the rabies section at the bottom Clearly identify as a rabies suspect and clearly indicate if human exposure has occurred with the route of exposure and date included A referring veterinarian must be listed on the form The submission form can be downloaded from http www sdstate edu vs adrdl forms index cfm 4 Samples arriving to the laboratory before 12 PM noon will have results available the same day Samples arriving after 12 PM noon will be tested the next business day 5 Additional tests if requested will not be performed on a rabies suspect case until the rabies FA has been completed and is negative See NEUROLOGIC DISEASES for submission recommendations 6 The ADRDL is open 8 AM to 5 PM Monday through Friday excluding holidays A SPECIMEN DROP OFF COOLER is accessible to the public 24 hours a day so samples can be delivered to the lab on
38. ens aseptically with the same care as for bacteriologic specimens 2 It is most important that specimens for virologic studies be kept cold after collection and during shipment Freezing is not necessary unless there will be a long delay before reaching the laboratory and it may interfere with some of the tests 3 Rabies refer to the Rabies section under Laboratory Policies whenever possible submit the entire animal together with a complete history of the circumstances If this is impractical remove the head at the atlanto occipital joint Caution The cerebrospinal fluid and cord may be infectious Place the entire head in a double plastic bag or container which will not leak Label Rabies and attach the history to the container and place in a transportation box DO NOT OPEN THE HEAD SAMPLING amp SUBMISSION GUIDELINES TISSUES If in doubt send some of each of the major body organs Put the tissues in sterile plastic bags and chill BLOOD For BVD isolation from live cattle take 10 ml of blood for serum or 10 ml of unclotted blood EDTA purple top tube chill Maternal antibody may interfere with detection of BVD virus isolation Therefore pre colostral blood is recommended for detection of BVD in persistently infected calves Blood from calves at 4 5 months of age is suitable Do not use Venoject or PST tubes For PRRSV information see the PRRS Virus page on the SDRDL website FECES A minimum of 2 ml of fecal mater
39. er or the submitting veterinarian It is the responsibility of the veterinarian to report the laboratory results to the owner In addition if veterinarians would like to receive the final results by e mail and or fax please contact us at 605 688 5171 to personalize your account Password protected preliminary and final case results are available to the veterinarian via the Internet refer to the VADDS REPORT GENERATOR at http www sdstate edu vs LIVE ANIMAL SUBMISSION Do not bring animals into the receiving office During regular business hours please come into the receiving office so that we may complete a submission form and help you to unload your animal s Large animals live and dead may be unloaded at the receiving dock east side of the ADRDL during regular business hours For customer convenience necropsy samples serum and small dead animals can be left in the ADRDL cooler 24 hours day 7 days week It is the responsibility of the client to adequately identify such samples and leave a completed submission form Tags and submission forms are located in a mailbox just outside the necropsy cooler On the weekends after regular business hours you will need to contact the veterinarian on call at 605 690 1576 to have large animals unloaded CARCASS DISPOSITION Carcasses become the property of the ADRDL and cannot be returned to the submitter Carcasses are incinerated or rendered according to local state and federal regulations
40. erimental assays only please contact the lab for more information BETA HYDROXYBUTYRATE BHA BHA is one of three main ketone bodies produced by the rumen microflora and utilized by the liver and other tissue for energy Ketosis and elevation of serum BHA results when the production and absorption of ketone bodies exceeds their utilization Together with elevated BHA and other ketone bodies acetone and acetoacetic acid there is elevated NEFA and decreased serum glucose during clinical ketosis The BHA assay by itself is recognized as the most sensitive determinant of subclinical ketosis during early lactation in dairy cattle INTERPRETATION OF BHA VALUES FOR OUR LABORATORY IS AS FOLLOWS Normal lt 10 mg dL Suspect subclinical 10 to 25 mg dL ketosis Probably subclinical 25 to 40 mg dL ketosis Clinical ketosis gt 40 mg dL NONESTERIFIED FATTY ACIDS NEFA NEFA or long chain fatty acids are the main transport form of lipid between fat stores in adipose tissue and the liver where they are used in the synthesis of triglycerides Cattle in late stages of pregnancy are actively mobilizing these fat stores in preparation for lactation as a supplement to nutritional sources An elevation in serum NEFA in the context of time before calving indicates that the animal may be receiving inadequate nutrition during this period Excessive NEFA mobilization may also overwhelm the first steps of triglyceride synthesis in the liver and set the an
41. esults is unexpected testing again 4 to 12 weeks later by an alternative procedure such as virus isolation or PCR is recommended purple top EDTA blood tube submitted for buffy coat culture or red top serum tube for virus isolation Producers may wish to separate ear notch positive animals from their herd mates if they choose to re test positive animals OTHER TESTS AVAILABLE A list of all immunohistochemistry tests performed at SDSU is provided in the charts for infectious agents and cellular biomarkers at http ihc sdstate org A variety of infectious agents including bacteria viruses and fungi can each be identified in situ in tissues from numerous species For example ADRDL uses antibodies to Bovine Viral Diarrhea Virus BVDV to immunohistochemically detect BVD in visceral tissues and skin ear notches In many cases autolysis of tissues prevents making the diagnosis with other tests such as a fluorescent antibody FA test When quick same day results cannot be obtained with FA testing immunohistochemistry IHC may be used to provide a more rapid diagnosis usually 4 Page 28 to 7 days after the sample is received than tests such as viral isolation VI which require 2 weeks or longer Similar to tumor pathology in humans veterinary medicine can now use species specific e g anti goat anti rabbit anti human etc cross reacting antibodies to help determine tumor cell type and or to identify metastatic site
42. fee of 50 sample If the serum sample is to be held any length of time it should be frozen a good deep freeze or freezer over refrigerator will be sufficient Do not freeze whole blood samples or samples with the clot remaining Hemolysis should be prevented since it interferes with interpretation of many serologic tests The following don t list will help Don t use wet needles syringes or tubes Don t freeze whole blood do not over expose to direct sunlight or heat Don t use chemicals for cleaning or disinfecting tubes syringes needles etc Detergents and other chemicals are a source of toxicity in serum neutralization tests Don t force blood through a needle remove the needle and then depress the plunger Care must be taken in collection and handling of swine and canine blood since it hemolyzes easily Antibodies to many viral infections are common in animal populations If you are submitting serum samples for diagnosis i e bovine respiratory disease acute and convalescent samples are imperative Most infections require 10 14 days to develop demonstrable antibodies in the serum The acute sample should be obtained in the early stages of disease Identify each sample on the tube not on the stopper Send a completed history form with the samples use a separate form for each herd or owner Avoid dumping samples into boxes or bags since the possibility of breakage occurs and or additional time is spent sorting sample
43. ferring veterinary clinic if one is listed will also be notified Page HOW TO SUBMIT RABIES SUSPECT CASES TO THE SOUTH DAKOTA PUBLIC HEALTH LABORATORY 1 Call the South Dakota Department of Health to report the possible exposure and to seek guidance in how to submit the animal for testing Call 1 800 592 1861 or 605 773 3737 during regular business hours For emergencies after hours on weekends or holidays call the mobile phone 605 280 4810 Staff will be able to answer questions and concerns If at all possible please call before destroying the suspect animal 2 Call one of the above numbers to make special arrangements for shipping an animal specimen after regular business hours on weekends or holidays 3 Notify the South Dakota Public Health Laboratory SDPHL of all impending shipments of animal specimens before actual transport Call the lab at 1 800 592 1861 or 605 773 3368 during regular business hours After hours on weekends or holidays contact an individual listed in 1 4 Be careful not to destroy the head of the animal by gunshot or bludgeoning Take the animal to a veterinarian for removal of the head in order to preserve the brain tissue and prevent unnecessary exposure to a diseased animal 5 Include with the specimen a SDPHL submission form with the following information Name and birth date of person exposed or owner if pet exposure Type of animal and exposure including exposure date suspect an
44. for sample submission as leakage or contamination might occur Samples received at the ADRDL lab in Gloves or Whirl Pack Bags may be assessed an extra handling fee to cover extra labor involved with sample processing CULTURE PLATES Cultures submitted for identification must be pure cultures Plates arriving at the laboratory with mixed bacterial growth will not be examined further Page SEROLOGY GENERAL SAMPLING PROCEDURES 1 Collect blood samples in clean sterile tubes avoid Bang s tubes which are not sterile Brucellosis tubes are NOT acceptable for submitting serum for serologic testing except for brucellosis testing Vacutainer type tubes are preferred Fill the tube to full a full tube is difficult to separate from the clot Blood samples from poultry are best collected with a syringe and serum transferred to a tube for shipping Allow the blood to stand undisturbed at room temperature for several hours to permit a clot to form and retract If using separation tubes i e SST or Corvac be sure to spin tubes properly for complete separation before shipping the silicon gel will interfere with sample pipetting and test results Avoid subjecting serum with the clot to temperatures below freezing or above 85 F When using snap capped tubes be sure lids are snapped on tight It is recommended that serum be removed from the clot and refrigerated Serum samples submitted on the clot will be charged an additional processing
45. gross lesions are present submit fresh chilled and formalin fixed samples Bronchial lymph node Fresh chilled lymph nodes augment bacterial or viral examinations Lung Depending on the size of the animal submit the entire lung one side or generous portions of lung fresh chilled Do not freeze and do not add glycerin Take samples of pneumonic lung adjacent unaffected lung and at the interface of pneumonic and normal lung Submit formalin fixed thin slices lt 1 cm of lung from affected and adjacent unaffected areas A slice through the affected unaffected interface is helpful Page RABIES Animals suspected of having rabies that have exposed a human should be euthanized and tested as soon as possible and staff at the ADRDL is qualified to perform the needed rabies FA test Since the FA test is so quick and reliable after hours testing is rarely required anymore however ANY AFTER HOURS WEEKEND OR HOLIDAY EMERGENCY RABIES TEST should be directed to the South Dakota Public Health Laboratory 615 East 4 st Pierre SD 57501 See below HOW TO SUBMIT RABIES SUSPECT CASES TO ADRDL To meet CDC guidelines for rabies testing the ENTIRE BRAIN WITH BRAINSTEM must be submitted FRESH to the laboratory This will allow for testing of both sides of the brain and brainstem as per CDC guidelines ADRDL staff will fix the brain from domestic animals and some wild animals in formalin for histopathology examination after rabies testing has be
46. he automated count Also includes hematocrit and erythrocyte parameters plasma protein and fibrinogen BLOOD COUNT An abbreviated exam that includes total counts for leukocytes erythrocytes platelets hemoglobin and erythrocytes parameters performed on the automated blood analyzer PRESURGICAL EVALUATION An abbreviated exam that includes manual hematocrit plasma protein and fibrinogen BLOOD TYPING Rapid testing to define major compatibility groups in dog DEA1 1 and cat A B AB samples in order to assess suitability for transfusions Coomb s testing may be required for dog samples in some instances COOMB S TEST Tube agglutination test for the presence of anti erythrocyte autoantibodies in dog serum URINE FLUIDS CYTOLOGY URINALYSIS Submitted urine samples are characterized by specific gravity pH and standard semi quantitative biochemical tests The sample is then centrifuged and the sediment is examined microscopically and described FLUID ANALYSIS Other fluids such as joint taps CSF and bronchoalveolar lavage can be characterized in a similar fashion relying mainly on protein determination pH and cytologic examination of the centrifuge pellet or cytospin prep of the sample KETONE TESTING IN FLUIDS A rapid colorimetric test is available to detect approximate ketone acetoacetic acid levels in body fluids such as milk urine and peritoneal samples CYTOLOGY However samples arrive at the lab cy
47. he cells and causes hemolysis Hemolyzed serum yields questionable results selenium and iron values will be falsely elevated and other minerals may also be affected Page FOOD SAFETY MICROBIOLOGY MEAT INSPECTION PROGRAM TESTING The ADRDL Food Safety Microbiology section is responsible for testing meat and environmental samples from meat processing plants that fall under the South Dakota and North Dakota State Meat Inspection Programs As both states adhere to US Department of Ag Food Safety Inspection Service directives methods selected for the FSM section are given preference if they appear in the sae de Microbiology Laboratory Guidebook The guidebook is available online at For this reason the method selected for screening samples for Salmonella Escherichia coli 0157 H7 Listeria spp and Listeria monocytogenes is the DuPont Qualicon BAX system BAX is a PCR based rapid method with results typically available 5 hours after sample enrichment is complete The following tests are available to personnel from the South Dakota Animal Industry Board Meat Inspection Program and the North Dakota Department of Agriculture Meat Inspection Program These tests may also be requested by ADRDL pathologists in special situations MEAT INSPECTION PROGRAM TESTING ACCEPTABLE SCREENING CONFIRMATORY SAMPLE SET UP SAMPLES METHOD METHOD DATES Salmonella Ground beef Carcass sponges BAX PCR Culture M Th Ready to eat meats
48. hipped in a small amount of mineral oil within a vial please do not send mineral oil samples in plastic bags Skin scraping samples or hair is best shipped dry in a sealed vial plastic bag or sealed paper envelope For submission of unusual samples or other requests please contact the lab Page TEST INTERPRETATION LABORATORY NORMAL RANGES FOR CLINICAL CHEMISTRY based on normative data for each species collected and performed by ADRDL or from applicable published data PARAMETER UNITS CANINE FELINE EQUINE PORCINE BOVINE OVINE TP g dL 5 7 7 8 6 8 8 4 6 4 8 1 6 8 6 7 8 7 6 0 7 9 ALB g dL 2 5 3 7 2 3 3 0 3 8 2 3 2 6 3 7 2 4 3 0 GLOB g dL 2 7 4 4 4 5 2 3 2 4 0 5 3 6 3 3 5 5 0 3 5 5 7 GLU mg dL 60 115 70 130 68 107 57 100 37 75 50 80 BUN mg dL 10 28 16 32 14 24 5 18 10 20 8 20 CRE mg dL 0 6 1 5 0 8 1 8 1 2 2 0 0 5 2 3 0 8 1 9 1 2 1 9 URI mg dL TBIL mg dL 0 15 1 00 0 1 1 0 0 5 2 4 0 1 0 8 0 2 0 7 0 1 0 5 DBIL mg dL 0 15 0 60 0 1 0 65 0 5 2 0 0 1 0 7 0 2 0 7 0 1 0 4 FE LJg dL CHOL mg dL AP IU L 10 133 12 67 76 220 ALT IU L 15 94 20 100 AST IU L 16 59 16 60 193 405 12 70 50 125 264 350 GGT IU L 6 24 10 60 8 0 29 0 20 52 AMYL IU L 247 799 400 890 LiP IU L LDH IU L CK IU L 100 325 284 933 58 409 65 209 NA mmol L 135 155 141 157 130 144 133 150 128 145 139 152 K mmol L 4 0 5 4 3 9 5 6 2
49. hopping the leaves ruptures the cells and releases the cyanide Freezing and thawing liberates cyanide from the sample rendering analysis useless Therefore it is best to submit samples promptly in a sealed plastic bag without freezing Page 60 Analysis of the suspected source of cyanide usually feed or plants is by far the most dependable economical and fastest method of diagnosis However in extreme cases we can try to test for cyanide in tissues preferred submission include stomach content blood heart blood brain and lung Tissue and blood specimens need to be frozen as soon as possible and analyzed within five days SALT POISONING OR WATER DEPRIVATION Causes sodium imbalance which can be detected by analyzing fresh brain tissue and serum SELENIUM HAIR For long term or chronic exposure submit about 3 grams 1 cup full of clean flank hair The flank hair is continuously being shed and will most accurately reflect the more recent selenium level in the animal Mane or tail hair which is not shed and therefore does not reflect recent selenium exposure BLOOD Blood reflects the exposure in the last two to three months Submit a minimum of 1 3 ml for analysis in an EDTA tube Blood must not be clotted SERUM and PLASMA Reflects the selenium exposure to the animal for about the last ten days UREA AND NONPROTEIN NITROGEN FEED ADDITIVES Suspected feed is the best submission for diagnosis of toxicosis due to urea and n
50. http www sdstate edu vs Page 8 LOCATION amp SUBMISSION INFORMATION Highway 14 Bypass Exit 132 Animal Disease Research And Diagnostic Lab N Campus Drive GZ MESA Day Arepoy Sixth Street Highway 14 From Interstate 29 take Exit 133 Turn west onto Highway 14 Bypass and travel to first stoplight Turn south onto Medary Ave At the first opportunity get into the left hand turn lane and turn left east onto North Campus Drive ADRDL is the second building in the left north side of the road Look for the diagnostic lab receiving sign on the east side of the building Submissions are received from 8 00 12 00 noon and from 1 00 5 00 p m Monday through Friday with the exception of state and federal holidays Emergency diagnostic services may be arranged on evenings weekends and holidays for bona fide emergencies This should be pre arranged by contacting the lab before closing or after hours by contacting the on call diagnostician at 605 690 1576 The emergency service is not provided for delivery convenience or procrastination For customer convenience necropsy samples serum and small dead animals can be left in the ADRDL cooler 24 hours day 7 days week It is the responsibility of the client to adeguately identify such samples and leave a completed submission form Tags and submission forms are located in a mailbox just outside the necropsy cooler Mail is picked up on Saturday and placed in the co
51. ia should be maintained at room temperature until incubated Please label the pouches in numeric order 1 2 3 instead of the bull name or number This is a more efficient and quicker way for our lab to make sure that all the pouches have arrived safely You can either reference pouch number with bull number on the submission form or keep track of which number corresponds to which bull at your clinic ex 1 Freddie The ADRDL provides InPouch TF pouches to practitioners at cost we buy pouches in large lots and can make testing slightly less expensive with this service Pouches have an expiration date of 1 year if stored at room temperature Please refer to the Current Fee Schedule for costs You can either incubate and check the pouches yourself or return them to the lab and we will incubate and examine If you return them you will be billed for the examination plus case accession charges and other fess Laboratory fees are subject to change without notice If performing your own incubation and examination the pouches should be incubated upright at 98 6 F If no trichomonads are detected on day 6 they are considered negative Page PATHOLOGY BIOPSY SERVICE IN order to ease submission of surgical biopsy specimens and to shorten turn around time for diagnosis the ADRDL provides reusable returnable shipping containers to veterinary practices This new ready to use shipper is designed for smaller specimens such as biopsy cyto
52. ial rectal swabs do not provide enough fecal material for EM examination EM is not suitable for detection of BVD virus in feces of carrier animals NASAL SWABS Swab deep into the nasal cavity For FA examination for viruses swabbing must be vigorous enough to collect epithelial cells Keep the swabs moist and refrigerated Direct examination for swine influenza virus is available by an ELISA format SCABS Collect scabs from mammillitis lesions and healing vesicles in sterile plastic bags chill VESICLES Aspirate any fluid present with a sterile syringe and needle Collect affected skin in a sterile plastic bag chill TONSILS For diagnosis of pseudorabies and cholera in swine collect both tonsils in sterile plastic bags chill BRAIN If CNS signs are involved submit the brain as instructed under the section entitled RABIES Pack all specimens in an insulated container with enough ice packs to maintain refrigeration until the specimens reach the laboratory Page CATTLE Adenovirus BRSV BVD Herpesvirus 2 Herpesvirus group 4 DN599 IBR Parvovirus PI 3 Pox virus CANINE Adenovirus 1 Coronavirus Herpesvirus AGENTS DETECTABLE BY ISOLATION SWINE SHEEP Adenovirus Adenovirus Circovirus Border Disease virus Enterovirus EMC PI 3 virus Herpesvirus pseudorabies Influenza virus Parvovirus PRRS Reovirus FELINE Calicivirus Feline Herpesvirus Other viruses not listed above can be isolated in
53. imal death date Symptoms and or unusual behavior of suspect animal Name and phone number of veterinarian or physician Submission forms are available from veterinarians or physicians 6 Wrap animal head carefully and either ship or deliver directly to the lab in an insulated container with ice or ice packs SPECIMEN MUST NOT BE FROZEN Transport the specimen by the quickest means possible 7 Direct additional questions to the SDPHL at 605 773 3368 Rabies information from the SD Public Health Lab is below and additional information can be found on their website at http doh sd gov Lab rabies aspx For additional information see the Compendium of Animal Rabies Prevention and Control 2008 http www cdc gov mmwr preview mmwrhtml rr5702a1 htm The Centers for Disease Control and Prevention http www cdc gov ncidod dvrd rabies default htm Page 21 SCRAPIE SPECIMENS TO SUBMIT 3 1 Fresh unfixed retropharyngeal lymph node 2 Fresh unfixed pharyngeal tonsil For details about how to collect the retropharyngeal lymph node and the location of the pharyngeal tonsil see http agbiopubs sdstate edu articles ExEx11012 pdf 3 Fresh unfixed brainstem amp obex A pictorial obex removal is available at http www aphis usda gov vs nvsl BSE procedure manual pdf Page TRICHOMONIASIS Culturing for Tritrichomonas fetus infection in bulls and cows has become a more common procedure with recent regulatory conce
54. imal up for fatty liver syndrome after parturition This test is generally applied clinically to dairy cattle normal ranges have not been established for beef breeds or other ruminants Above normal NEFA levels at a given stage indicate that body fat is being mobilized rapidly and the cow is probably losing more weight than she should at that stage of gestation or lactation NORMAL NEFA VALUES FOR OUR LABORATORY ARE gt 2 weeks before calving lt 0 32 mmol L 2 weeks before or after calving lt 0 40 mmol L gt 2 weeks after calving lt 0 60 mmol L Page BOVINE IGG1 ASSAY A radial immunodiffusion RID assay is performed to measure IgG1 subtype levels in serum of neonatal calves as a sensitive indicator of colostrum absorption and failure of passive transfer FPT INTERPRETATION OF IGG1 VALUES FOR OUR LABORATORY IS AS FOLLOWS Failure of passive lt 800 mg dL transfer Partial passive transfer 800 1600 mg dL Adequate passive gt 1600 mg dL transfer These ranges apply to beef and dairy calves 1 to 8 days of age at time of sample collection As calves age normal ranges for serum IgG1 drop slightly so that by 60 days of age the low normal value is 1100 mg dL PARASITOLOGY AND FECAL ELISA TESTING FECAL FLOTATION Detection of protozoal cysts and helminth ova based on fecal flotation High density flotation may be performed if trematode infection is suspected Semiquantification scoring of parasite burden is reported in
55. ished as per AAVLD standards are followed from the time we receive samples to reporting results to the clients as rapidly as we can Page 6 COMPUTER BASED CASE REPORTING VADDS The Animal Disease Research and Diagnostic Laboratory SDSU Brookings South Dakota offers access to your verified laboratory results through the World Wide Web Our Internet address is http www sdstate edu vs To obtain your veterinary clinic s personalized security code and password please complete the form below and return it by mail to the address below or fax the completed form to 605 688 6003 ADRDL SDSU Box 2175 North Campus Drive Brookings SD 57007 1396 VETERINARY CLINIC NAME VETERINARIANS CURRENTLY AT THE CLINIC ADDRESS CITY STATE ZIP CODE PHONE FAX VETERINARY CLINIC S AUTHORIZATION SIGNATURE pO Cid If a password change is required at a later date please contact our office 605 688 5171 or 605 688 6003 fax ADRDL Office Use Only User Code Password Effective g Page 7 CONTACT INFORMATION MAILING SHIPPING ADDRESS Animal Disease Research and Diagnostic Laboratory Department of Veterinary Science North Campus Drive Box 2175 South Dakota State University Brookings SD 57007 1396 605 688 5171 Main Office 605 688 6003 Fax FACULTY CONTACTS http www sdstate edu vs faculty index cfm WEB PAGE
56. itis Western Blot Forwarded to Equine Diagnostic Solutions Kentucky All samples must be received by 10 30 a m or they are not done until the next working day Be prepared to allow up to 4 weeks before receiving results from National Veterinary Services Laboratory NVSL Ames IA The National Veterinary Services Laboratory requests that serology submissions include an acute and convalescent serum sample Samples forwarded to other diagnostic laboratories by the South Dakota Animal Disease Research amp Diagnostic Laboratory are mailed M W Page 36 OVINE SEROLOGIC TESTS AGENT TEST PURPOSE INTERPRETATION Bluetongue AGID ELISA Bovine Viral Diarrhea Virus or VN Detect infection Positive 4 fold rise in antibody titer between paired sera Border Disease Antibody titer gt 1 256 usually significant Brucella ovis ELISA Detect infection Positive suspect and negative determined by relative OD value as compared to controls Johne s Para tuberculosis ELISA Detect infection Calculation based on sample OD Ovine Progressive Pneumonia AGID Detect infection Positive Precipitin line Negative No precipitin line Toxoplasmosis Agglut Detect exposure amp Positive Sheep gt 1 64 infection IFA SCHEDULE FOR OVINE SEROLOGIC TESTS TYPE OF TEST Bluetongue AGID On demand Same day ELISA Bovine Viral Diarrhea Virus or Border Disease VN i Johne s Para tuberculosis C Ovin
57. l buffered formalin Please do not use whirl pak bags for the ear notch test Number the red top tubes 1 2 and 3 etc Write the animal identification number that corresponds to each tube number on the submission form please print carefully Do not write the animal identification number directly on the tube Tattoo ink will not interfere with the test and the sample does not have to be kept under sterile conditions SHIPPING THE SAMPLES If the ear notch is stored in 10 formalin for more than 5 or 6 days false negative results may occur Therefore please ship the samples as soon as they are collected The tissue will undergo fixation while the samples are in route to the diagnostic lab RESULTS Results will be available in approximately 10 14 days during the busy spring season sooner during other times of the year INTERPRETATION The BVD ear notch test is a screening test designed to detect persistently infected cattle By tradition persistently infected cattle have been defined as those cattle that remain infected with the virus for longer than 4 weeks usually indefinitely Normal cattle that are not persistently infected will typically clear the infection within 4 weeks time It is believed that the vast majority upper 90 percentile of IHC ear notch positive animals are persistently infected with BVD virus and many veterinarians will recommend culling after one positive test If the animal is highly valued or the rest r
58. late antigen test CF Complement fixation test ELISA Enzyme linked immunosorbent assay HI Hemagglutination test IFA Indirect fluorescent antibody test IH Indirect hemagglutination MA Microagglutination test RSAT Rapid slide agglutination test 2M RSAT 2 Mercaptoethanol rapid slide agglutination test VN Virus neutralization Page 39 INTERPRETATION OF SEROLOGIC TEST RESULTS You must use great care in interpreting the results of serologic tests especially on single serum samples At best serologic reaction to an infectious agent is indirect evidence that the animal was exposed to the agent in the past Attempts to associate a current clinical condition with a serologic reaction are full of pitfalls While it is necessary to designate specific titers to define infected and non infected animals in disease control and eradication programs everyone should recognize that these titers are set arbitrarily and that they are not truly definitive There is no one antibody titer for any infectious disease that always differentiates infected from non infected animals The following factors must be considered when using serologic tests as aids in diagnosis THE FREQUENCY WITH WHICH THE INFECTION OCCURS IN THE POPULATION Some organisms e g porcine parvovirus bovine viral diarrhea virus IBR virus PI 3 virus BRSV etc cause undetected infections so frequently that it often is difficult to find mature animals without antibodies to these agen
59. leak proof containers and on cold packaging e The PCR assay is set up once weekly on Thursday PRRSV NORTH AMERICAN EUROPEAN EUROPEAN LIKE SCREENING ASSAY e Specimen Serum Raw Semen Extended Semen Tissue Blood Swabs Oral Fluids e Submit all labeled samples in leak proof containers and on ice packaging e Samples may be pooled per client request recommended limit of 5 per pool however pooling may decrease the sensitivity of the test e Please submit separate aliquots for additional tests i e Serology Virology e The PCR assay is set up 5 days a week PRRSV NORTH AMERICAN EUROPEAN EUROPEAN LIKE QUANTITATIVE ASSAY e Specimen Serum Raw Semen Extended Semen Tissue Oral Fluids Blood Swabs Page 45 e Submit all labeled samples in leak proof containers and on ice packaging e For quantitation of tissues gram weight of tissue submitted is needed eg tissue should be submitted rather than homogenate where weight is unknown e The PCR assay is set up 5 days a week e Positive PCR results will be reported in copies ml for fluid samples and copies gram for tissue samples SWINE INFLUENZA SIV NAHLN National Animal Health Laboratory Network Surveillance e Specimen Lung or nasal swabs from pigs showing influenza like illness or associated with other pigs showing influenza like illness e Testing for presence of Influenza A is within 1 business day of receipt e Virus isolation with subsequent sequencing perfo
60. logy and clinical pathology submissions It contains a small jar with formalin for a biopsy sample a protective slide mailer for cytology submission packing materials instructions and applicable forms When the shipper is received by the lab the box is unpacked and returned to the submitter with replacement items included Cytologic specimens can be examined in a variety of ways depending on sample characteristics site of collection and potential diagnoses If there is an aspirate fluid at least 0 5 mL the sample should be placed into an appropriate sized EDTA tube and sent to the lab on ice do not freeze These samples will be tested by fluid analysis Smaller samples can be spread onto slides and left unstained then shipped to the lab Sending stained slides to the lab is less ideal since everyone s stain methods are different and reading an atypical stain will make interpretation a problem Unstained slides may be shipped in the protective slide mailer provided with the biopsy shipping box Cytology samples can be submitted with either the Biopsy Submission Form or the Clinical Pathology Submission Form EXCISIONAL BIOPSIES ADRDL STANDARD BIOPSY SERVICE The ADRDL standard biopsy service includes histologic examination of up to three masses from the same animal up to two special stains as indicated for diagnosis and return of the biopsy shipping container with fresh formalin at a reduced laboratory fee see appendix for current
61. m phosphate monobasic monohydrate 4gm Sodium phosphate dibasic anhydrous 6 5 gm Specimens for formalin fixation should not be more than 0 5 cm thick If the tissue is more than 0 5 cm thick it needs to be sectioned such that adequate fixation can occur cut bread slices approximately cm apart in the tissue leaving the bottom of the specimen intact so that tissue orientation is not lost Your specimen should look something like this Figure 1 This type of sectioning ensures that adequate fixation will occur when the tissue is placed in 10 X the volume of 10 neutral buffered formalin and lesion orientation will not be lost since the specimen remains intact SHIPPING The use of screw cap wide mouthed containers for formalin fixed tissue is preferred Plastic containers are better than glass jars for shipping samples to the lab Do not use narrow mouth containers fixed tissue swells and cannot be removed from the container Do not submit large specimens multiple specimens of partially fixed or unfixed tissues in small shipping volumes of formalin the tissue will continue to undergo autolysis and putrefaction if there is insufficient formalin for complete tissue fixation histologic examination of the specimen s may be impossible Once the tissues are fixed approximately 24 hours for necropsy cases you may pour off most of the formalin leaving just enough liquid to cover the tissue this will reduce the fee
62. n 10 buffered formalin Colon or Cecal Content fresh e 2 5 mlin plastic bag or tube Mesenteric lymph nodes one each from mid and lower gut fresh FOR OPTIMAL ENTERITIS DIAGNOSTIC SUCCESS Samples must be taken as soon after death as possible Do not open or tie off intestinal tissue that you are submitting to the lab Do not freeze Pool all formalin fixed tissues in one container with adequate amounts of formalin For fresh tissue package and label the small intestine separately from the cecum colon No not mix intestinal samples with other viscera Chill fresh intestine and colon content before mailing Pack in an insulated diagnostic shipping container with enough ice packs to maintain refrigeration until the specimens reach the laboratory Page NEUROLOGIC DISORDERS EXCLUDING RABIES Many toxic nutritional and metabolic causes of CNS disease do not produce lesions in the brain and require analysis of other tissues samples for definitive diagnosis submission of whole blood EDTA rumen contents feed sample s water sample s liver kidney and brain may aid in diagnosis of many of these causes ROUTINE SPECIMENS FOR SUBMISSION NOTE if the brain is being submitted for rabies examination the ENTIRE brain should be submitted fresh chilled see Rabies section BRAIN Brain fresh chilled including brain stem Do not fix we will place tissue in formalin after rabies testing if needed is performed ENTIR
63. oid the necessity of re bleeding animals we will not accept serum submitted in brucellosis tubes for tests other than brucellosis 12 Serum samples submitted on the clot will be charged an additional processing fee of 50 sample Submission of serum on the clot is a significant cause of poor samples received at the laboratory These samples reguire additional technical time and material and cause delays in testing of the serum This charge will cover labor and materials expended in the preparation of serum Direct any guestions regarding serum preparation and or collection tubes to Serology Department SDSU ADRDL Phone 605 688 5171 Page BOVINE SEROLOGIC TESTS AGENT TEST PURPOSE INTERPRETATION Anaplasmosis Detect Card Test Infection Positive Agglutination Cervidae Negative No agglutination cELISA Positive gt 30 PI Negative lt 30 PI Bluetongue AGID Detect Positive Precipitin line infection Negative No precipitin line ELISA Results based on sample OD Bovine Leukemia Virus AGID Detect Positive Precipitin line infection Negative No precipitin line do not test the month before or the month after parturition ELISA Color change Bovine Respiratory VN Detect Positive 4 fold rise in antibody Syncytial Virus infection titer between paired sera Negative No antibody or less than 4 fold rise Antibody titer gt 1 128 may indicate recent infection in unva
64. oler for processing on Monday Mailed specimens should be sent through the fastest practical carrier Mailed submissions should be timed to avoid weekend and holiday delivery delays whenever possible A completed submission form must accompany all submissions The submission form is intentionally designed to be short and concise The information requested is necessary to focus the examination and thereby limit the time and cost of laboratory testing A minimal amount of information is requested for epidemiological purposes Your cooperation in filling out the submission form as completely as possible is greatly appreciated and will enhance the quality of our diagnostic investigation Page 9 LABORATORY POLICIES RECEIVING 8 BUSINESS HOURS Monday Friday 8 00 a m 5 00 p m Except on Holidays After hours emergencies are referred to aveterinarian on call at 605 690 1576 SUBMISSION FORMS Submission forms can be downloaded at http www sdstate edu vs adrdl forms index cfm VETERINARY REFERAL 8 REPORTING e Mammalian cases should be submitted by or on referral from a licensed veterinarian e Poultry are accepted from flock owners or poultry servicemen e Rabies specimens are accepted from veterinarians physicians and public health officials e State and federal wildlife personnel may submit wildlife Final signed reports and invoices will be mailed to the submitting veterinarian and can be sent to the owner upon the request of the own
65. on protein nitrogen feed additives Abnormal levels of ammonia and urea may be detected in the stomach or rumen contents heparinized whole blood or serum if these samples are collected immediately after death and are frozen immediately after collection and remain frozen during transportation to the laboratory This is another exception to the general rule of never freezing whole blood samples Autolysis greatly decreases usefulness of analysis of the specimens out of the animal A set of tissues in formalin should also be sent to the laboratory VITAMIN E AND VITAMIN A Submit a minimum of 1 5 ml of frozen serum Vitamin E degrades very rapidly and the serum must be kept frozen from time of collection until analysis SUSPECTED POISONOUS MATERIALS Samples of any suspected materials such as chemicals plants paints drugs rubbish or old batteries should be submitted to aid in confirming the source of a toxic agent Packaging will depend on the sample Be sure to consider any necessary safety precautions which might be required for proper handling and shipping Page TRACE MINERALS TRACE MINERALS A minimum of 4 ml of serum is needed for a complete mineral screen Blood needs to be spun down and the serum drawn off before the sample is shipped or frozen Plastic snap top tubes are preferred for shipping the serum Red rubber top tubes may affect the zinc values Do not use serum separator tubes Freezing of whole blood ruptures t
66. ontact surface swabs BAX PCR M W Environmental Swabs Generic E coli ok ok Caleass swabs 3M E coli Coliform Petrifilm Plate M W Food Carcass swabs Sample must arrive at the laboratory within 24 hours of collection and arrive at the lab at a temperature of 32 F to 40 F Campylobacter j c I Carcass swabs Direct Count M F Yeast Mold Count Food 3M Yeast Mold Count Petrifilm Plate M F Aerobic Plate Count Food 3M Aerobic Plate Count Petrifilm Plate M W SUBMISSION GUIDELINES e Holiday sample submission procedures will be determined on a case by case basis Visit http www sdstate edu vs foodsafetymicrobiolab index cfm for the latest holiday food safety submission schedule e Insulated shipping containers and ice packs should be used as appropriate Page APPENDICES CURRENT FEE SCHEDULE QUALITY MANUAL SUBMISSION FORMS Page 65
67. rmed to determine Hemagglutinin H Neuraminidase N and Matrix M type approximately 3 weeks TRITRICHOMONAS FOETUS e Specimen In Pouch TF Presumptive Positive genital fluid and tissues e The PCR assay is set up once weekly on Monday e Direct PCR can be performed on smegma sample submitted in 5 mls of saline depending on export requirements SEQUENCING SERVICES AVAILABLE DNA SEQUENCING e Specimen PRRS or PCV2 PCR Positive RNA samples are used e The expected turn around time is about 5 to 7 days upon receipt of the sequencing request or viral isolate Page CLINICAL PATHOLOGY The Clinical Pathology section of ADRDL performs routine and reference veterinary hematology clinical chemistry parasitology and fluid analysis from clinical samples For details on parameters included in combination tests or profiles please refer to the submission form appendix XI E Beyond tests listed additional methods can be performed either as special methods or through collaborating labs please contact us for more information HEMATOLOGY COMPLETE BLOOD COUNT Evaluation of white red and platelet cell populations through use of a laser based automated blood analyzer Cell Dyn 3500R Abbott Laboratories supplemented with manual examination of the blood smear to describe cell morphology confirm automated cell counts and perform white cell differential Unless otherwise stated the manual differential will be reported rather than t
68. rns A hallmark of Tritrichomonas testing at ADRDL is strict adherence to OIE World Organization for Animal Health guidelines The OIE publishes among other documents a Manual of Diagnostic Tests and Vaccines for Terrestrial Animals that is recognized by the World Trade Organization as rules for international reference OIE Tritrichomonas testing guidelines include USE OF CULTURE POUCH INPOUCH TF This is an OIE endorsed culture media that also serves as transport media Samples that will not arrive at the lab within 24 hours of collection should be submitted in transport media for optimum sensitivity Use of the InPouch TF negates the need for separate transport media Samples are incubated and examined directly in the pouch there is no need for technicians to manually transfer culture media to a slide for examination The entire pouch is examined under microscopy ensuring that any trichomonads in the culture are detected The smegma sample is deposited directly into the pouch eliminating the need for transfer to another container before shipment to the lab CULTURES EXAMINED AFTER APPROPRIATE LENGTH OF TIME OIE guidelines call for examination of culture media up to 7 days after inoculation normally day 6 post arrival at the lab At the ADRDL positive Tritrichomonas cultures have not been noted prior to 4 days incubation at the earliest when the original inoculum was smegma from the bull Attempts to read results earlier therefore
69. s Allow sufficient time for completion of serologic tests some tests can be completed on the day of arrival and others take up to 7 days Serological tests are done on a set schedule some tests are done daily whereas others are only set up once or twice a week The schedules are on the following pages Please call when submitting a large number of samples gt 200 to insure that the laboratory has sufficient reagents to meet your needs Page 32 10 11 Ship samples in an insulated container with sufficient ice packs to maintain refrigeration until arrival at the laboratory Special shipping containers for blood tubes are available for purchase at the laboratory call 605 688 5171 for more information Brucellosis tubes are NOT acceptable for submitting serum for serologic testing except for brucellosis testing With the introduction of more sophisticated and sensitive tests for serology serum quality becomes very important for accurate and reproducible results Many ELISA tests and all serum neutralization tests require clean serum In addition hemolysis interferes with the interpretation of complement fixation tests used for anaplasmosis and Brucella and hemagglutination tests for swine influenza and parvovirus Brucellosis blood collection tubes are frequently contaminated with cleansing chemicals and or bacteria making interpretation of test results difficult In order to facilitate the rapid turnover of test results and av
70. s in lymph nodes In veterinary medicine there is an on going need to correlate tumor identification and tumor biology with information about the patient s prognosis and response to specific treatment protocols In the future veterinary medicine will use immunohistochemistry to stage tumor development determine the patient s prognosis and evaluate patient response to treatment ANTIGEN ANTIBODY REACTIONS 8 IMMUNOHISTOCHEMISTRY To be observable with a light microscope antigen antibody reactions require that the antibody molecules must be labeled with a compound that permits their visualization in the cells Although many different protocols are used depending on the antibody the typical technique used at ADRDL involves the application of a primary antibody followed by a secondary labeling antibody detection system Positive controls are derived from non patient tissues known to express the antigen of interest Other cell types known to express the antigen of interest within the patient tissue sample may also serve as an internal positive control Negative controls are patient tissues which are not exposed to the primary antibody but are otherwise processed the same as positive control tissues Also patient tissue expected to be negative i e no expression of the antigen of interest are examined to verify that the primary antibody did not cross react with non specific antigens in the patient Please call 605 688 5171 or email Tanya
71. sampled SAMPLES REQUIRED FOR SPECIFIC TOXINS Identification of different toxins requires examination of specific samples or organs Page ALKALOIDS Such as strychnine nicotine monocrotaline and various drugs are usually identified chemically in urine kidney liver and stomach contents Histopathologic examination of formalin fixed tissue is helpful in confirming diagnosis ANTICOAGULANTS When sweet clover poisoning or anticoagulant rodenticides are suspected whole blood including blood found in the body cavities liver and baits should be submitted Secondary samples include kidney stomach content and feed Formalin fixed tissues should be sent CARBAMATES Some carbamate pesticides such as carbofuran Furadan are very toxic while another carbaryl Sevin is of relatively low toxicity Stomach contents feed or suspected poison source are the preferred samples FLUORIDES If chronic fluoride toxicosis is suspected submit urine bone teeth feedstuffs and water A rib bone is a good sample If acute fluoride toxicosis is suspected handle like METALS see below HERBICIDE FUNGICIDE The specimens needed to aid in the diagnosis of herbicide or fungicide poisoning are stomach contents liver kidney feed and other suspected sources of poison METALS When checking for poisoning by metals such as lead arsenic or copper kidney liver urine and stomach content are preferred Unclotted blood EDTA or
72. sh brain in a sterile plastic bag and place inside a crush proof container keep brain refrigerated chilled until it reaches the laboratory Section formalin fixed brain transversely not longitudinally once before placing in formalin to aid in fixation if brain is large package in crush proof container Entire heads should be chilled not frozen prior to shipping and shipped in an insulated leak proof container with enough ice packs to maintain refrigeration until specimen reaches the laboratory When in doubt concerning sample submission contact the laboratory Page PNEUMONIA RESPIRATORY DISEASE SUBMISSION SUGGESTIONS The nasal cavity trachea bronchial lymph nodes and lungs should be considered in animals with respiratory disease Nasal cavity Swabs They should be long enough to reach deep into the nasal cavity Swabs to be used for virus FA examinations must penetrate the mucus layer to retrieve epithelial cells Check the information that comes with the swabs many swabs may not be suitable for bacterial or viral examinations due to additives Swabs must be kept moist and cool before and during shipment Snout or turbinate Formalin fixed turbinate can be examined microscopically This can be very helpful for checking for Inclusion Body Rhinitis in pigs Tonsil In pigs it is helpful in examination for viral respiratory pathogens to submit the tonsil half in formalin and the other half fresh and chilled Trachea If
73. should be milked out by pressing the tip of the swab between the fingers through the flexible wall of the pouch LOADING THE INPOUCH TF SYSTEM Insert the pipette tip containing the specimen through the pouch opening and empty contents into the liquid of the upper compartment The 0 5 to 1 cc of smegma material is forced out of the pipette with the syringe Do not overfill the system or bacterial proliferation will ruin the test If the collected material adheres to the wall of the pipette it can be rinsed by drawing a small amount of medium into the pipette and then flushing back into the compartment The production of bubbles should be avoided to maintain the anaerobic conditions of the medium To close the pouch fold and roll the top of the bag down 2 or 3 times and then fold tabs behind the pouch to lock Next squeeze contents of the upper compartment into the lower compartment Roll down the pouch until the closure is at the top of the liquid with the label still visible and fasten tabs as above The loaded system should be maintained between 59 F and 98 F until transported to an incubator During the warm months samples can usually be safely shipped to the lab in an insulated box without ice and then be incubated upon arrival Remember do not refrigerate or freeze the specimen OTHER COMMENTS If culturing for both T foetus and Vibrio campylobacteriosis place of the specimen in the InPouch and into the Vibrio media Both med
74. st animals do not need to be opened longitudinally adequate fixation occurs from both the mucosal and serosal surfaces provided the intestinal lumen has been flushed with formalin BRAIN NOTE IF RABIES TESTING IS DESIRED DO NOT FIX BRAIN IN FORMALIN SUBMIT ENTIRE BRAIN FRESH CHILLLED Brain should be cut in half longitudinally and one half placed in a wide mouth container of 10 neutral buffered formalin The cerebrum of large animals hogs cattle horses sheep bison etc should be bread sliced see Figure 1 prior to submersion in 10 neutral buffered formalin to allow adequate tissue fixation Entire small animal brains cats dogs and neonates of most species can be placed directly into a wide mouth jar containing 10X the volume of 10 formalin Handle brain gently to prevent the cerebellum from becoming detached and include the obex brain stem whenever possible Page IMMUNOHISTOCHEMISTRY BVD EAR NOTCH TEST Collect ear notches using a pig ear notcher or similar tool Be SURE that each notch consists of skin and not just strands of hair If you can feel the tool cut through the cartilage of the ear you have correctly notched the ear Avoid collecting tissue from the tip of the ear or any traumatized areas that are covered with a scab and may consist of ulcerated skin only Place each ear notch in a separate 10ml red top serum tube or similar sized centrifuge tube Fill each tube with 7 to 8ml of 10 neutra
75. tologic accessions are evaluated by a pathologist as an aid to diagnosis Cytologic specimens can be examined in a variety of ways depending on sample characteristics site of collection and potential diagnoses If there is an aspirate fluid at least Page 47 0 5 mL the sample should be placed into an appropriate sized EDTA tube and sent to the lab on ice do not freeze These samples will be tested by fluid analysis Smaller samples can be spread onto slides and left unstained then shipped to the lab Sending stained slides to the lab is less ideal since everyone s stain methods are different and reading an atypical stain will make interpretation a problem Multiple unstained slides if possible optimize the pathologist s flexibility in examining the sample LEUKOCYTE LINEAGE ANALYSIS Using a panel of standard cytochemical stains lymphoproliferative and myeloproliferative disorders can be classified according to analogs of FAB nomenclature as an aid to prognosis and therapy Please contact the lab for more information prior to sending samples Page SERUM CHEMISTRY Tests are available either individually as standard profiles based on species or clinical problem or as customized profiles With the exception of thyroid hormone and cortisol testing all assays are performed with an automated serum chemistry analyzer VetAce Analyzer Alfa Wasserman Please contact the laboratory for more information INDIVIDU
76. ts Obviously you must use great care in using single sample serologic test results for diagnosing such infections THE LONGEVITY OF THE SEROLOGIC REACTION CAUSED BY THE INFECTION Some infectious agents cause high antibody levels that may persist for many years For example sheep infected with Toxoplasma gondii will maintain significant antibody titers 2 or more years after recovery from the acute stage of infection The antibody titer and how long it persists depends on the agent the species infected and the differences in immunologic reactions of individual animals Once an animal has been immunized to an agent each subsequent exposure to the agent by vaccination or natural exposure may elicit an anamnestic response that results in an increase in the level and persistence of the titer THE POSSIBILITY OF IMMUNOTOLERANCE Some infectious agents e g BVD virus produce immunotolerance rather frequently While other agents may not cause true immunotolerance infection by them may not always induce the production of antibodies detectable by the usual tests For example up to 25 of cows infected by serovar hardjo of leptospira have no antibody detectable by the microagglutination test Heifers born to cows infected with Brucella abortus may have a latent infection but have no antibodies detectable by the usual tests until near or after parturition Thus the absence of antibody is not always assurance that infection does not exist VARIATIONS I
77. ue specimens and labile organisms may be lost entirely during transit to the laboratory ANTHRAX Blood or spleen samples are preferred for anthrax diagnosis Clotted blood drawn from the dead animal can be used Other than for Bacillus anthracis blood is a poor sample for bacteriologic examination BRUCELLA INFECTIONS Slaughter specimens for Brucella culture should be collected immediately shipped to the laboratory frozen CLOSTRIDIAL INFECTIONS Specimens to be examined for clostridia should be collected immediately after death and kept well preserved to prevent contamination with nonpathogenic spore forming organisms ENTERIC INFECTIONS On enteric cases in piglets calves and lambs four air dried ileal impression smears should be sent for testing for E coli pili For identifying enteric pathogens in live animals fecal samples are preferred over rectal swabs This is especially true for isolation of Salmonella At least a gram of fecal material should be submitted for Salmonella culturing Salmonella isolates are sent to the National Veterinary Services Laboratory in Ames lowa for serotyping The results will be sent to the veterinarian approximately 4 weeks after the case report JOHNES SUSPECTS Call the Bacteriology lab 605 688 5171 before submitting 50 or more fecal samples for culture Collect samples carefully in wide mouth leak proof screw cap or snap top hard plastic containers Please do not use Gloves or Whirl Pack Bags
78. y o Enrichment cultures for Salmonella require an extra day o Campylobacter cultures require 3 5 days Tissues for microscopic examination are trimmed on the day following receipt and are returned to the pathologists on the afternoon of the third working day One or two days are required to read the slides and dictate the letters o It takes 1 2 days to deliver mailed reports to the submitting veterinarian m Some serology tests are not set up daily because of the small number of samples or because of the time requirements of some tests please refer to the serology schedule for additional information TEST SET UP SCHEDULE For specific information about how often tests are set up and expected turn around times click on the TEST SET UP SCHEDULE tab at http www sdstate edu vs adrdl index cfm Page SPECIFIC SYNDROMES ARTHRITIS SPECIMENS TO SUBMIT ANTEMORTEM Joint fluid Use aseptic technique a sterile needle and syringe to draw joint fluid POSTMORTEM Entire joint The limb may be severed above and below the affected joint and the entire joint submitted Put entire joints into plastic bags and seal Joint fluid Use aseptic technique a sterile needle and syringe to draw joint fluid or use aseptic technique to open the joint with a sterile blade Use a sterile swab to reach deep into the joint cavity Put containers of joint fluid and joint swabs into plastic bags and seal Ship in an insulated container with enough ice pa
79. you pay for shipping due to the reduction in total package weight The small size of most biopsy specimens allows fixation to occur during shipping You do not have to wait 24 hours before shipping the specimens Caution if you are using a whirl pak bag as your formalin container do not twist the two ends together like a twist tie on a loaf of bread the formalin WILL leak Figure 2 ROUTINE HISTOPATHOLOGY COLLECTION OF SPECIMENS Collect representative tissue s and or lesions associated with the suspected disease Process and include adjacent normal tissue whenever possible If no gross lesions are apparent submit specimens from all major body organs including brain NECROPSY CASES Place specimens for histologic examination in a labeled container of 10 neutral buffered formalin containing 10 times the volume of formalin to the volume of the specimen s ORGANS Tissues such as liver kidney spleen heart and lung should be sliced into 0 5 cm thick specimens Fix these organs in an adequate volume of 10 neutral buffered formalin If tissues are extremely congested and bloody double the volume of 10 formalin INTESTINE 3 cm long segments of duodenum jejunum ileum spiral colon and colon should be gently flushed with 10 neutral buffered formalin prior to submersion This ensures that the intestinal content is removed and the mucosal surface is exposed to formalin Sections of gastrointestinal tissue from mo

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