Bovine Disease Diagnostic Manual FIRST EDITION A FIELD GUIDE TO DIAGNOSING COMMON BOVINE DISEASES Editor: David H. Zeman, DVM, PhD, DACVP Assisting the cattle industry in solving challenging herd health problems using cutting-edge technology and personalized customer support. Custom Made Vaccines Table of Contents As the nation’s largest manufacturer of custom made vaccines, Newport Laboratories is a highly focused, technology–based company dedicated to providing timely, science–based solutions to food animal disease problems. We do this by providing customers with industry-leading science backed by PINPOINT® Technologies. Our products and services are delivered and supported by a dedicated and experienced sales staff and Veterinary Technical Service team. Copyright Newport Laboratories, Inc. All rights reserved. Table of Contents Bovine Diagnostic Submission Guide Pg 4 Tissue Submission Guidelines 6 Major Bovine Organs 9 Bovine Necropsy Instructions 10 Respiratory Diseases Histophilus somni Pneumonia 12 BRSV 14 Calf Diphtheria 15 Infectious Bovine Rhinotracheitis (IBR) 16 Mannheimia Bronchopneumonia 17 Mycoplasma Pneumonia 18 Pasteurella Pneumonia 19 Atypical Interstitial Pneumonia (AIP) and Acute Bovine 20 Pulmonary Emphysema and Edema (ABPEE) Syndromes Gastrointestinal Diseases BVD 22 Coccidiosis & Cryptosporidiosis 23 Clostridium perfringens Diseases 24 Johne’s Disease 26 Enteric Colibacillosis 27 Salmonellosis 28 Viral Enteritis of Neonates 29 Multisystemic Diseases Septicemic Colibacillosis 30 Histophilus somni TEME & Myocarditis 12 Salmonellosis 28 Miscellaneous Diseases Actinomycosis (Lumpy Jaw) 31 Footrot - Fusobacterium necrophorum 32 Hardware Disease 33 Liver Abscess - Fusobacterium necrophorum 34 Polioencephalomalacia 35 Pinkeye - Moraxella bovis/bovoculi 36 Pyelonephritis 38 www.newportlabs.com | 800-220-2522 3 | Bovine Diagnostic Submission Guide Disease Suspected Specimen Sample Preparation Laboratory Procedure Abscessed Tissue 10% Formalin Histopathology & Tissue Stains Actinomycosis Abscessed Tissue & Exudate Fresh, Refrigerate Anaerobic Bacterial Culture Anaerobic culture swab Refrigerate Anaerobic Bacterial Culture Refrigerate PCR, Virus Isolation Bovine Herpes Virus 1 Lung, Trachea, Nasal Swabs (IBRV) 10% Formalin Histopathology Refrigerate PCR, Virus Isolation Bovine Respiratory Lung Syncytial Virus (BRSV) 10% Formalin Histopathology Lung, Ileum, Cecum, Spiral Refrigerate PCR, Virus Isolation Bovine Virus Diarrhea Colon, Mesenteric Lymph Nodes, (BVDV) Spleen, Whole Blood (Purple top 10% Formalin Histopathology tube) Lesion Material, Swab from Refrigerate Anaerobic Bacterial Culture Calf Diphtheria Lesion Caseous Lymphadenitis Purulent Exudate from Abscess Refrigerate Culture-sensitivity (sheep & goats) Anaerobic Culture, PCR, Colon (sections with contents) Refrigerate Clostridium difficile A/B Toxin ELISA Small Intestine (tied off), Anaerobic Culture-sensitivity, Toxin Refrigerate Clostridium perfringens Abomasum (fresh), Liver (fresh) PCR Testing Types A, C, D, E Note: Do not submit fecal samples 10% Formalin Histopathology Coccidiosis/ Small Intestine (with Cecal & Refrigerate Fecal Float Cryptosporidiosis Spiral Colon & Contents), Feces 10% Formalin Histopathology Small Intestine (Jejunum, Ileum, Refrigerate Culture & K99 Pilus Identification Colibacillosis & Colon), Feces from calves less than 7 days old. 10% Formalin Histopathology Lung, Liver, Spleen, Kidney, & Fresh Bacterial Culture Lesions Colisepticemia Lung, Liver, Spleen, Kidney, & 10% Formalin Histopathology Lesions Coronavirus Lung, Nasal Swabs Refrigerate PCR, Virus Isolation (respiratory) Intestine (with Cecal & Spiral PCR, Virus Isolation, Sequencing Enteritis (Rotavirus Colon Section & Contents) Refrigerate and Coronavirus) Feces PCR, Virus Isolation, Sequencing Footrot (Fusobacterium Swab from Lesion Refrigerate Culture, PCR Subspeciation necrophorum) Hardware Disease Reticulum, Pericardium 10% Formalin Histopathology For bacterial culture, we recommend swabs with transport media to prevent desiccation. For virus isolation, swabs should be placed into viral transport media; call us for information. FREE Diagnostic Shippers • 800-220-2522 • www.newportlabs.com | 4 Newport Laboratories | Bovine Disease Diagnostic Manual Bovine Diagnostic Submission Guide Disease Suspected Specimen Sample Preparation Laboratory Procedure Culture-sensitivity, Gene Refrigerate Sequencing, PCR for Affected Lung Immunoglobulin Binding Proteins 10% Formalin Histopathology Histophilus somni Culture-sensitivity, Gene (formerly Haemophilus Refrigerate somnus) Myocardium, Brain Sequencing, PCR 10% Formalin Histopathology Culture-sensitivity, Gene Joint Swabs (if involved) Refrigerate Sequencing, PCR Lung, Ventral & Dorsal Areas Fresh Bacteriology & Virology Interstitial Pneumonia Lung, Ventral & Dorsal Areas 10% Formalin Histopathology Ileum 10% Formalin Histopathology, Acid-Fast Stain Johne's Disease Ileum or Feces Fresh, Refrigerate Johne's Culture, PCR Culture-sensitivity, Gene Mannheimia Refrigerate Affected Lung, Lymph Nodes Sequencing, PCR, Serotyping haemolytica (formerly (mediastinal or bronchial) Pasteurella haemolytica) 10% Formalin Histopathology Milk Sample from Affected Refrigerate or Frozen Culture-sensitivity Mastitis Quarter or Bulk Tank Culture, Speciation, Gene Refrigerate Affected Lung Sequencing 10% Formalin Histopathology Mycoplasma bovis Joint Swabs/Fluids, Culture, Speciation, Gene Tracheal Swabs, Swabs or Refrigerate Sequencing Exudate from Middle Ear Refrigerate PCR, Virus Isolation Parainfluenza-3 Virus Lung, Nasal Swabs (PI-3) 10% Formalin Histopathology Affected Lung, Lymph Nodes Refrigerate Culture-sensitivity, PCR Pasteurella multocida (mediastinal or bronchial) 10% Formalin Histopathology Pinkeye (Moraxella Culture-sensitivity, PCR Speciation bovis, M. bovoculi & M. Eye Swabs Refrigerate (virulence factors), Gene ovis) Sequencing Cerebral Cortex, Brainstem 10% Formalin Histopathology Polioencephalomalacia Water, Feed/Forage Fresh Sulfate Analysis Kidney, Urinary Bladder 10% Formalin Histopathology Pyelonephritis Kidney, Urinary Bladder, Urine Fresh, Refrigerate Bacterial Culture Reproductive failure/ Fetus with Placenta if Available Refrigerate Reference Lab abortion Small Intestine, Spleen, Lung, Culture-sensitivity, Gene Refrigerate Liver, Mesenteric Lymph Nodes: Sequencing Salmonellosis (all fresh and in formalin;) Fresh Feces & Fecal Swabs 10% Formalin Histopathology Trace Mineral Deficiency Liver (fresh) Refrigerate Reference Lab (i.e. copper & selenium) Newport Laboratories | Bovine Disease Diagnostic Manual www.newportlabs.com | 800-220-2522 5 | Tissue Submission Guidelines Whenever possible, animals selected for laboratory analysis should be free from antibiotic therapy and in an early or acute disease stage. Selected tissues should be collected as aseptically as possible. A meaningful history of the disease outbreak and a tentative diagnosis, based upon clinical evaluation and necropsy findings, should be included. Laboratory test results are directly affected by animal selection, necropsy technique, specimen selection and specimen handling, including preservation and shipment to the laboratory. Contact Newport Laboratories if you have any questions regarding sample collection or the diagnostic process. Preparation & Collection of Tissues/Samples Tissues-Fresh Nasal Swabs-Viral Suspect Aseptically collect approximately 2x4 inch samples Prepare nostrils and sample as in bacterial suspect. ® and place in a plastic bag. Sample visible lesions with For viral swabs use Viral Culturette (Becton Dickinson adjacent normal tissue. Double bag in Whirl-pak® bags. #4361514) or equivalent. Do not mix swabs, intestines, or brains with other Use of the incorrect swab and media may jeopardize tissues in one single bag. Transport tissues with 2-3 the ability to detect or culture the offending pathogen. cold packs in an insulated container. It is important that For bacterial isolation, avoid using Mycoplasma sp. or the tissue samples arrive at the laboratory before the viral media which contain antimicrobials and may inhibit cold packs expire. growth of the desired pathogen. Avoid using bacterial Collect sections of small and large intestine. The culture media to isolate viruses or Mycoplasma sp. selected, clearly identified samples should be double organisms. bagged and sealed in Whirl-pak bags to prevent spillage. Do not longitudinally cut the loops of Identify all swabs with the following: intestines open. The intestine, approximately 2 inches • Farm ID, including site and pasture/lot long, should be refrigerated and cooled thoroughly where appropriate prior to shipping. Avoid shipping over the weekends • Animal identification number or holidays. Histopathology Swabs Preparation of Tissue for Fixation Aerobic Culture Multiple sites or types of lesions, to include both Commercial swabs with Stuart’s or Amies transport normal and diseased tissue and a sample at the line of media is recommended to prevent desiccation. demarcation, should be taken. The sections should be no more than 1 inch thick. The small size of the Anaerobic Culture tissue results in rapid and complete penetration of Port-A Cult® (BBL) or other anaerobic transport system. the fixative. (The Port-A Cult® tube can be used for anaerobic, facultative, and aerobic bacteria.) For abscesses or Selected tissues should be cut with a sharp knife or exudates use a capped syringe with needle removed or scalpel since the squeezing action of scissors crushes a tube with a snug cap.