LABORATORY FOR CLINICAL DIAGNOSTICS
Directory of tests 2019/20 2019/20 Foreword LABORATORY FOR CLINICAL DIAGNOSTICS Dear colleagues, you are now holding our catalogue, very Our fields of interest: conventional, indeed, as a paper version, First things first - taking care of the but also available as a digital one to analytics to serve your needs, to serve browse online. So many lab manuals exist. your clients, is our first and foremost Still, we believe we have put together goal. At the same time, whenever we find some useful information that is not so the existing analytics lacking something, easily at hand, a very unique manual either accuracy or speed or availability, indeed. we will look into that in detail. Research has always been part of our work to gain Our lab profile: access to innovative new methods or You will find some basic parameters tests. You can rely on us! and profiles as well as some unique parameters that are not so frequently Our service: found elsewhere but very helpful. We take From sample collection to availability of pride in establishing new tests, always results 24/7 - we are convinced that it is searching for the best possible technique, our duty to make analytics and the work the best possible parameter. Some with the lab as easy as possible. Invoicing specialties: You will find a vast range of is either by order or on a monthly basis. tests especially created to meet the needs And turnover dependent debates are in the diagnostic field of small mammals something we give regularly, no need and reptiles. And our antibiotic testing to contact us. Plus, our representatives is taylor-made to help with an optimised throughout the European countries will be therapy for all species involved. happy to help either by phone, fax or by paying a visit whenever a problem arises. Our quality: Just let us know please! Accreditation ensures that the test quality We want you to be able to concentrate on is constant and certified by external diagnostics and therapy - just leave the specialists. You as our customer can lab part to us! rely on that. Accreditation audits the qualification of the people involved as Our vision: well. You can be assured, our system In an ever-changing world there is a of constant learning is set up to make constant need for accuracy and reliability innovation possible within our lab. The for the vet when commissioning the lab. growing number of specialists, including We promise to give you the best possible diplomates in various fields and members support. of university staffs working for us makes You can rely on us! it possible for us to take an active part in congresses and seminars throughout Kind regards Europe. At the same time, we love routine diagnostics and we are happy to discuss cases with you in practice. Dr. Elisabeth Müller CEO LABOKLIN GmbH & Co KG
3 2019/20 LABOKLIN at a Glance 2019/20 Contents LABORATORY FOR CLINICAL DIAGNOSTICS LABOKLIN at a Glance Contents LABOKLIN is accredited according to DIN EN ISO/IEC 17025:2005 Foreword 4 2.1.4 Blood Donor Profiles – Small Animals 39 An overview of LABOKLINs range of services LABOKLIN at a Glance 4 2.1.5 PCR Profiles – Dog/Cat 40 Profiles and Screenings Pathology 2.1.6 Faecal Profiles – Contents 5 • Dog and cat • Histopathology Small Animals 42 • Small mammals • Immunohistology Our contacts 8 2.2 Profiles – Small Mammals, • Birds • Cytology Birds and Reptiles 43 • Reptiles • BRAF Mutation Abbreviations 11 2.2.1 Clinical Chemical • Horse • Exsudate/Transudate Profiles 43 1 Pre-analytics 13 • Ruminants • Cerebrospinal fluid 2.2.2 PCR Profiles – 1.1 Blood, plasma, serum samples 13 • New World camelids • Synovia Small Mammals, • Pig • Other aspirates 1.1.1 Preparation of the Birds and Reptiles 45 • Amphibians patient 13 2.2.3 Faecal Profiles – PCR detection 1.1.2 Which sample? 13 Small Mammals, Blood examinations • Dog 1.1.3 Factors interfering with • Allergy • Cat analysis 15 Birds and Reptiles 47 • Endocrinology • Small mammals 1.1.4 Specific features 17 2.3 Profiles/Screenings – Horse 48 • Function tests • Birds 1.2 Microbiology 17 2.3.1 Clinical Chemical • Haematology • Reptiles 1.3 Hygiene 18 Profiles 48 • Clinical chemical parameters • Horse 1.4 Histology and immuno 2.3.2 PCR Profiles – Horse 52 • Serology/Infectious diseases • Cattle histochemistry 18 2.3.3 Faecal Profiles – Horse 53 • Pig 1.4.1 Skin punches 19 2.4 Profiles/Screenings – Hereditary diseases • Amphibians 1.4.2 Cytology 19 Ruminants 54 • Dog • Fish 1.5 Polymerase chain reaction 2.4.1 Clinical Chemical • Cat • Sex determination in birds (PCR) 20 Profiles 54 • Horse • etc. 1.6 Genetic testing 20 2.4.2 Serological Profiles – • Cattle 1.7 Immune status 22 Ruminants 56 • Pig Other genetic examinations 1.8 Sample material/Shipping 2.4.3 PCR Profiles – • etc. • Breed analysis material 22 Ruminants 56 1.9 Labelling 24 • Identity and parentage 2.4.4 Faecal Profiles – 1.10 Packaging and transport 26 Hygiene • DNA profile Ruminants 57 • Hygiene examinations • Species differentiation 1.11 Reordering tests 27 2.5 Profiles/Screenings – Pig 58 • Profiles • Coat colour/coat structure 2 Profiles and Screenings 29 2.5.1 Clinical Chemical Profiles 58 2.5.2 Serological Profiles – Pig 59 Microbiology and parasitology Preanalytics 2.1 Profiles/Screenings – 2.5.3 PCR Profiles – Pig 59 • Bacteriology • Blood, plasma, serum samples Companion Animals 29 • Mycology • Microbiology 2.1.1 Clinical Chemical 2.5.4 Faecal Profiles – Pig 60 • Virology • Urine samples Profiles 29 2.6 Profiles – Hygiene 60 • Parasitology • Histology und immunohistology 2.1.2 Serological Profiles 36 3 Haematology 61 • Maldigestion/Malabsorption • Polymerase chain reaction (PCR) 2.1.3 Travel Profiles and 3.1 Blood Cells 61 • Autovaccines • Immune status Thrombocyte Profiles • etc. Small Animals/ 3.2 Coagulation 63 Tick-borne Diseases 37 3.3 Blood Grouping 66
4 5 2019/20 Contents 2019/20 Contents LABORATORY FOR CLINICAL DIAGNOSTICS
4 Clinical Chemistry 68 15.2 Testing for Specific Parasitic or 20.3.2 Coat Colour/Coat 23.4 Reference ranges farm 4.1 Enzymes 68 Protozoa Infections 244 Structure Cat 326 animals 358 4.2 Substrates 74 15.3 Parasitological Examination – 20.4 Horse 328 23.4.1 Clinical chemistry 358 4.3 Minerals and Trace Minerals 80 Skin 245 20.4.1 Hereditary Diseases 328 23.4.2 Haematological reference 20.4.2 Coat Colour/Coat ranges farm animals 359 5 Urinalysis 86 16 Tests for Indigestion and Structure Horse 335 Diarrhoea 246 20.4.3 Performance Horse 338 24 Conversion Table for Laboratory 6 Allergy 90 16.1 Bacteriological Examination 246 20.5 Cattle 339 Diagnostic Parameters 360 6.1 Allergy Testing 90 16.1.1 Faecal Profiles 246 20.5.1 Hereditary Diseases 339 24.1 Clinical-chemical Parameters 360 6.2 Allergen-specific 16.1.2 Testing for Specific 20.5.2 Breed Characteristics 24.2 Blood Parameters 361 Immunotherapy 95 Indigestion/Diarrhoea Cattle 342 25 Courier Service 362 Pathogens (Bacteria) 250 20.6 Small Ruminants and 7 Immunological Tests/Markers for 16.2 Virological Examination 252 New World Camelids 343 26 Invoicing 362 Inflammation 96 16.2.1 Faecal Profiles – Virology 20.6.1 Hereditary Diseases 343 252 8 Endocrinology/Tumour Markers 103 20.6.2 Breed Characteristics 16.2.2 Testing for Specific Small Ruminants and Indigestion/Diarrhoea New World Camelids 345 9 Function Tests/Calculations 115 Pathogens (viruses) 253 20.7 Pig 345 16.3 Diagnostics of Maldigestion/ 10 Vitamins 127 Malabsorption 254 21 DNA Profile, Breed, Species 346 11 Drug Level 130 16.4 Determination of an Inflammatory 21.1 Identity and Parentage 346 Exudative Process 255 21.2 Breed and Species 347 12 Intoxication 132 17 Autovaccine/Herd-specific 22 Hygiene Examinations 349 13 Infectious Diseases: Pathogenic Vaccine 257 22.1 Profiles Hygiene 349 Agents and Antibody Detection 133 22.2 Single Tests 349 13.1 Viruses 133 18 Pathology 259 18.1 Histopathology 259 13.2 Bacteria 176 23 Reference Ranges 352 18.2 Immunohistology 259 13.3 Fungi 212 23.1 Dog, cat, horse 352 18.3 Cytology 260 13.4 Parasites 215 23.1.1 Clinical chemistry 352 18.4 Lymphocyte Clonality (PARR) 260 23.1.2 Haematological 14 Bacteriology/Mycology 234 18.5 BRAF Mutation 261 reference ranges 14.1 Smears/Puncture 18.6 Differentiation Exsudate/ dog, cat, horse 353 Transudate 261 Fluids/Milk/Faeces 234 23.1.3 Hormones dog, cat, 14.2 Skin/Hair/Feathers 237 19 Sex Determination in Birds 262 horse 354 14.3 Bacteriological Examination 23.2 Reference ranges rabbit, Horse 238 20 Hereditary Diseases/Phenotype/ guinea pig and ferret 355 14.4 Testing for Specific Infectious Breed Characteristics 263 23.2.1 Clinical chemistry 355 Agents 239 20.1 Heredities 263 23.2.2 Haematological 14.5 Sensitivity Testing 241 20.2 Dog 263 reference ranges rabbit, 14.6 Additional sensitivity testing 242 20.2.1 Hereditary Diseases 263 guinea pig, ferret 356 20.2.2 Coat Colour/Coat 23.3 Reference ranges birds 357 15 Parasitology 243 Structure Dog 312 23.3.1 Clinical chemistry 357 15.1 Parasitological Examination – 20.3 Cat 319 23.3.2 Haematological Faeces 243 20.3.1 Hereditary Diseases 319 reference ranges birds 357
6 7 2019/20 Our contacts 2019/20 Our contacts LABORATORY FOR CLINICAL DIAGNOSTICS Our contacts Laboratories Offices
LABOKLIN Germany Tel.: +49 (0) 971 7 20 20 LABOKLIN Belgium Tel.: +32 13 48 05 05 Steubenstraße 4 Fax: +49 (0) 971 6 85 46 E-Mail: [email protected] 97688 Bad Kissingen E-Mail: [email protected] LABOKLIN Croatia Tel.: +385 (0) 91 11 22 121 LABOKLIN Austria Tel.: +43 (0) 732 7172420 E-Mail: [email protected] Paul-Hahn-Straße 3 / BT-D / 1. Stock Fax: +43 (0) 732 717322 4020 Linz E-Mail: [email protected] LABOKLIN Czech Republic Tel.: +42 (0) 730 105 024 E-Mail: [email protected] LABOKLIN Nederlands Tel: +31 (0) 85 4890580 Verlengde Klinkertstraat 6 Fax: +49 (0) 971 68546 LABOKLIN Denmark Tel.: +45 4352 1228 6433 PL Hoensbroek E-Mail: [email protected] E-Mail: [email protected]
LABOKLIN Poland Tel.: +48 (0) 22 691 93 10 LABOKLIN Estonia Tel.: +372 5696 4488 ul. Powstańców Śląskich 101 Fax: +48 (0) 22 691 92 92 E-Mail: [email protected] 01-495 Warszawa E-Mail: [email protected] LABOKLIN Finland Tel: +358 (0) 40 / 7104017 LABOKLIN Slovakia Tel.: +421(0) 948 783 888 Tel: +358 (0) 44 / 0675353 Líščie údolie 57 E-Mail: [email protected] E-Mail: [email protected] 84231 Bratislava LABOKLIN France Tel: +33 (0) 9 67 32 85 80 LABOKLIN Spain Tel.: +34 644 030 557 Fax: +33 (0) 3 88 09 43 82 Polígono Industrial de Alcobendas Tel.: +34 91 198 67 71 E-Mail: [email protected] Avenida de la Industria 4, edificio 3 E-Mail: [email protected] Planta 1ª Oficina A LABOKLIN Iceland E-Mail: [email protected] 28108 Alcobendas (Madrid) LABOKLIN Ireland Tel: +353 19 02 68 06 LABOKLIN Switzerland Tel.: +41 (0) 61 319 60 60 E-Mail: [email protected] P.O. Box Fax: +41 (0) 61 319 60 65 4002 Basel (sample shipment) E-Mail: [email protected] LABOKLIN Italy Tel: +39 (0) 392 033 45 86 Max Kämpf-Platz 1, Erlenmatt, Fax: +39 (0) 051 082 19 75 4058 Basel E-Mail: [email protected]
LABOKLIN United Kingdom Tel.: +44 024 7632 3275 LABOKLIN Latvia E-Mail: [email protected] Batt Laboratories Ltd, E-Mail: [email protected] The Venture Centre University LABOKLIN Lithuania Tel.: +370 6122 2020 of Warwick Science Park E-Mail: [email protected] Sir William Lyons Road, Coventry CV4 7EZ 8 9 2019/20 Our contacts 2019/20 Abbreviations/additional information LABORATORY FOR CLINICAL DIAGNOSTICS
LABOKLIN Norway Tel.: +47 9946 2020 E-Mail: [email protected] Abbreviations Sample material LABOKLIN Portugal E-Mail: [email protected] You will find these abbreviations in part also on our submission forms. The materials required for the individual tests are also indicated on the submission forms, however, for LABOKLIN Russia E-Mail: [email protected] lack of space, not always completely.
LABOKLIN Slovenia E-Mail: [email protected] , Information connected with FACS fluorescence-activated cell comma: you can choose sorting LABOKLIN Sweden Tel.: +46 (0) 723 73 2020 which sample you want to FAVN fluorescent antibody virus E-Mail: [email protected] submit neutralisation + Information connected with FLP fragment length “+”: you must submit all of polymorphism these sample materials FTIR Fourier-transform infrared spectrometry A swab without medium HAH haemagglutination inhibition BAL bronchoalveolar lavage assay CB citrate blood HPLC high performance liquid CFS cerebrospinal fluid chromatography CP citrate plasma ICA immunochromatography EB EDTA blood assay EP EDTA plasma ICPMS inductively coupled plasma FA faeces mass spectrometry IFAT indirect fluorescent antibody H urine technique HB heparin blood ISE ion-selective electrodes HP heparin plasma LCMS liquid chromatography-mass NaFB sodium fluoride blood spectrometry S serum MALDI-TOF matrix-assisted laser TBS tracheobronchial secretion desorption/ionisation – time of TM swab with medium flight mass spectrometry MAT microscopic agglutination test Test methods PARR polymerase chain reaction for antigen receptor rearrange- AAS atom absorption spectrometry ments cELISA competitive ELISA PCR polymerase chain reaction CLIA chemiluminescence assay RIA radioimmunoassay CFT complement fixation test SAF sodium acetate formaldehyde EIA corresponds to ELISA STRs short tandem repeats ELISA enzyme linked immuno www.laboklin.com • www.labogen.com sorbent assay VNT virus neutralisation test
10 11 2019/20 Abbreviations/additional information concerning the test descriptions 2019/20 Pre-analytics LABORATORY FOR CLINICAL DIAGNOSTICS
Other abbreviations New World camelids llama, alpaca; 1 Pre-analytics AB antibodies Being polygastric, new world bdw body weight camelids may appear under * partner laboratory 1.1 Blood, plasma, serum samples the heading „ruminants“. The first step in the process of examining a sample is the pre-analysis. Pre-analysis Farm includes all steps from patient preparation, specimen collection and transport of the Numbers you may find in the test animals Ruminants and pigs sample to the lab to the preparation of the sample for analysis. descriptions (1) Specifications apply to testing 1.1.1 Preparation of the patient Further notes with method (1) Before taking a blood sample, the patient should normally fast for 10 - 12 hours, (2) Specifications apply to testing The obligation to notify the authorities upon provided the physiology of the species concerned permits this. Otherwise, faulty results with method (2) suspicion of a disease applies to Germany. are to be expected, especially for cholesterol, glucose and TLI. In addition, parameters (3) Specifications apply to testing The obligation to notify the authorities upon such as α-amylase, ALT, AST, bilirubin, total protein, triglycerides, serum bile acids, with method (3) diagnosis of a disease applies to Germany. leukocytes and calcium can be affected. Fasted blood samples are not unproblematic in horses and only indicated for special > greater than tests (e.g. insulin and glucose for the diagnosis of the Equine Metabolic Syndrome). Duration The information applies from < less than It is advisable to inform the owner about the influence of physical activity or stress on the date of arrival of the the results of a blood examination. Particularly muscular enzymes such as CK, LDH samples at Laboklin. and AST can show increased levels in the serum after physical exertion. Additionally, ”Days” means ”working days”. glucose and lactate can also show elevated serum levels. Due to delays in transport, test duration may take longer Before doing any allergy tests, including feedstuff tests, any administration of for tests which are carried out corticosteroids should be stopped. To do so, the following withdrawal times are by a partner laboratory. recommended: The specified test durations - local/topical corticosteroids: 2 - 4 weeks are supplied without liability. - oral corticosteroids (e.g. Prednisolone): up to 8 weeks - depot cortisone preparations (e.g. Voren®): up to 3 months If these times cannot be observed, false negative results are possible. In the case of a Species positive result, the reaction class must be assessed taking into account the previous Large administration of cortisone. Please note that other itch-suppressing medication may animals Horses and farm animals also have a negative impact on the allergy test. Our allergy team will be pleased to advise you. Small mammals Rabbit, guinea pig, rat, 1.1.2 Which sample? mouse, hamster, ferret and other small mammals which Details on the recommended material (blood, serum, plasma) for the requested test are being kept as pets can be taken from our test descriptions or the submission form. For labelling the In individual cases, tests sample, it is also necessary to indicate the sample type (see Chapter 1.9 and 1.10). for small mammals may be also applicable for small wild Whole blood samples mammals (e.g. hedgehogs). EDTA blood (EB) Small - For doing a blood count, EDTA blood is the most suitable material in mammals animals Dog and cat (however, for birds and reptiles it is heparin blood, see below). - For the serological examination of the blood type, EDTA whole blood is needed as well.
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- As the cells in the sample are not stable, EDTA samples for haematological tests Serum should not be older than 48 hours. - Samples are drawn into tubes without anticoagulants. - For most PCR analyses and genetic tests, EDTA blood is required. - Allow to stand for 30 – 60 min. - To determine certain parameters such as ACTH or pro-BNP, only EDTA plasma which - Centrifuge for 10 min at 2000 g. was promptly centrifuged and cooled can be used to obtain reliable results. - Remove the supernatant by pipette and transfer it into an uncoated test tube, then label the test tube. Heparin blood (HB) - For correctly determining individual parameters like insulin or fructosamine, only - To collect heparin samples, lithium heparin (LiHep) tubes are available. serum should be used. - For doing a blood count for reptiles and birds, lithium heparin blood should generally - Sending non-centrifuged samples should only be done exceptionally (e.g. in case of be used. a very low sample quantity), as the transport might result in cell damage and thus lead - Since the amount of blood is often very low in small mammals, lithium heparin tubes to a haemolytic serum. are particularly suitable, as they cannot only be used for doing a blood count but also for determining a wide range of blood-chemical parameters. - For the PCR, lithium heparin whole blood should only be used under exceptional 1.1.3 Factors interfering with analysis circumstances, as lithium heparin can inhibit the PCR and might thus lead to false negative results. Haemolysis Haemolysis is caused by leakage of intracellular components of the erythrocytes such Citrate blood (CB) as phosphate, iron, potassium and especially haemoglobin due to a damage of the cell - To determine the coagulation parameters, only the appropriate citrate tubes should be membrane. Haemoglobin causes a red colouration of serum/plasma which primarily used. For getting a correct evaluation, their shelf life may not be exceeded. It is also interferes with the photometric testing done in clinical chemistry. necessary to have an exact mix ratio of 1:10 (1 part citrate + 9 parts blood). - Most coagulation parameters can be analysed using citrate whole blood. Lipaemia - For correctly performing platelet function tests, citrate whole blood is required. Lipaemia refers to the milky/turbid discolouration of serum/plasma due to triglycerides. It is mostly caused by diet- or stress-related factors. Lipaemia can also occur as a result Sodium fluoride blood (NaFB) of endocrinological diseases like Cushing’s syndrome or hypothyroidism. Sodium fluoride inhibits enzyme activities which lead to a reduction of some parameters. It should be used for the correct determination of glucose and lactate. Lipaemic samples often complicate the measurement of certain clinical parameters, e. g. bilirubin. Plasma Icterus - Samples are drawn into tubes with anticoagulants (heparin, EDTA, citrate). Icterus is a yellowish discolouration of serum/plasma. Excess amounts of bilirubin, - Can be centrifuged immediately after collection (10 min, 2000 g). which is the reason for the yellow colouring, are normally caused by a medical condition - Remove the supernatant by pipette and transfer it into an uncoated test tube, then and cannot be influenced. indicate the sample materials on the test tube or use the appropriate bar code label. - Please note: The additives limit the number of analyses! The yellow colouration is physiological in horses. - Heparin plasma (HP) is needed for many clinical-chemical examinations. HP cannot be used for agglutination tests. - The collection of EDTA plasma (EP) for clinical-chemical and/or serological parameters should only take place in exceptional cases, as EDTA disturbs through various mechanisms the measurement of individual parameters such as calcium, magnesium and AP. Likewise, potassium cannot be determined when using EDTA plasma, since EDTA is added as K-EDTA. - Some coagulation parameters can only be analysed using citrate plasma (CP). Performing platelet function tests using centrifuged citrate plasma is not possible.
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Interfering factor Parameter Level 1.1.4 Specific features Haemolysis LDH, CK, AST, bilirubin, AP, creatinine, PO4, K, Mg, Blood counts Fe, fructosamine - EDTA or lithium heparin blood Haemolysis Ca, glucose - When collecting the sample, discard the first 0.5 ml of blood, if possible, as they contain an increased amount of coagulation factors, or first obtain a serum sample. Lipaemia ALT, AST, AP, bilirubin, glucose, Ca, PO4, - Let the blood run down slowly on the side of the sample tube. total protein, lipids, haemoglobin - Pay attention to the fill volume! Preferably fill up to the mark, since an insufficient Lipaemia albumin, amylase, Na, CI, K volume can result in changes in cell morphology. Do not overfill the tube in any case, as the sample might clot. Icterus AP, total protein, Cl, PO4 - After drawing the sample, tilt the test tube carefully several times. - Do not store blood smears in the refrigerator and not close to formalin. Icterus triglycerides, creatinine, Mg - Pack the samples frost-proof in winter; possibly cool them in summer. - Reliable results can only be obtained from samples not older than 48 hours.
Clinical chemistry of serum or heparin plasma Medicine Parameter Level Prompt centrifugation of the samples lead to better test results, as it reduces the risk of Penicillin G K haemolysis caused by transportation. However, serum should be allowed to stand for a minimum of 30 min to ensure a complete clotting of the sample. Tetracyclines PO4 Serum samples can also be shipped frozen; they will then reach the laboratory cooled. Repeated freezing/thawing, though, should absolutely be avoided. Tetracyclines K Determination of glucose and lactate Salicylates CK, AP, glucose, Na, total protein - Requires sodium fluoride blood or sodium oxalate blood Salicylates K, Ca - Fill the test tube up to the mark at the maximum. Corticosteroides CK, AP, glucose, Na, total protein Coagulation parameters - Determination is carried out using sodium citrate plasma which is obtained from Corticosteroides K, Ca citrate blood with the mix ratio being 10:1 (9 parts blood + 1 part sodium citrate). Centrifugation should take place at the practice. Phenylbutazone Ca, Na - If commercial citrate-treated tubes are used, the expiry date needs to be checked before collecting the sample. Expired tubes may no longer be used, as skewed results Barbiturates CK are to be expected. When drawing the sample, special attention must be paid to the Halothane CK, PO4 exact fill level (marking on the tube). anaesthesia - If no commercial tubes are available, sodium citrate 3.13% can be drawn into a syringe. - No heparinised needles or catheters may be used. Glucose infusion glucose Glucose infusion PO4 1.2 Microbiology
- It is important to collect samples as sterile as possible to avoid contamination with physiological and/or ubiquitous flora. - Smears for bacteriology (aerobic and anaerobic microbes) and for mycology should be sent with a transport medium (“swab with medium” = TM) to protect the microbes during shipment.
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- Smears for examinations using PCR should be sent without a transport medium Detailed explanations: (“smear without medium” = A, “swab without medium”, “dry swab”) or with a special As a sample, a representative piece of tissue free of preparation artefacts (e.g. medium. disruption, squashing, electrocoagulation) should be taken. The diameter of the sample - Urine samples should be sent in a sterile tube using Uricult or using a swab with should not be less than 0.5 cm. An exception to this are samples which, for technical transport medium. reasons, cannot be obtained otherwise (such as endoscopically taken stomach - Hairs and/or skin scrapings for the diagnosis of dermatophytes are best sent in a biopsies). Furthermore, it should be borne in mind that samples which are too small paper bag or in aluminium foil. only provide little information, whereas samples that are too large cannot be fixed - For sending faeces/excrement, special transport tubes should be used, no bags or properly. Pieces of tissue with an edge length of 1 cm are recommended. However, gloves tied with a knot; for birds, use cloacal swabs with transport medium. this might vary depending on the lesion to be examined, the sampling site and the - Blood culture flasks can be ordered with prior written notice (subject to a objective. charge). Small lesions should be placed centrally so they are not overlooked and thus truncated during preparation. If in doubt, several samples should be collected.
1.3 Hygiene 1.4.1 Skin punches As skin samples, punch biopsies of all dermal layers with a diameter ≥ 0.6 cm are to The contact plates required for examining the surface disinfection must be ordered be submitted. Primary lesions from several locations should be selected. The biopsied from the laboratory in good time, as they can only be stored for a short period of time. area should not be pre-treated by scraping or shaving. The anamnesis should contain all The surfaces to be sampled have to be cleaned and disinfected and must be well dried relevant data which might be important for the diagnosis. It is recommended to use our before sampling. The contact plates, which need to be labelled on the bottom, must be submission form Pathology, which especially focuses on skin and tumour diagnostics, returned cooled to the laboratory within 24 hours together with the filled-in submission but also leaves room for any other type of anamnesis. form. The bioindicators necessary for evaluating the proper functioning of the sterilisers 1.4.2 Cytology should be requested from the laboratory as needed and can be applied right in the Samples can primarily be taken as wipe test, scraping or puncture (with or without device together with the regular sterilisation material. After testing, these bioindicators aspiration). The most common technique is the fine needle aspiration, using a thin have to be sent directly to the laboratory together with the filled-in submission form. hollow needle (G 22 – G 27) attached to a syringe. A vacuum is created and, if possible, For testing the disinfection of endoscopes, two contact plates and two rinse samples the tissue should be punctured several times in different directions. Before detaching are necessary for each endoscope. Before testing, the material must be requested the needle, the vacuum must be released to avoid the material receding into the from the laboratory in due time. A detailed description of all the processes of taking the syringe. The material obtained is then pressed out of the needle onto the side of a glass sample will be provided. It is required to return the collected samples to the laboratory slide. A second slide is placed flat at a right angle on top of the first one and is then within 24 hours. carefully pulled away across the slide. If the sample is more liquid, a steeper angle (45°) – like in a blood smear – should be applied. For the cytological examination of aspirates, excretions or secretions, the fluids 1.4 Histology and immunohistochemistry obtained are centrifuged at 2500 – 3000 rpm for three to five minutes. The supernatant is decanted and the sediment is carefully spread like a blood smear and shipped air- When submitting tissue samples for histopathological and immunohistochemical exa- dried. If the aspirates are sent directly, EDTA tubes should be used as test vessels. minations, the following points must be observed: For bronchial, conjunctival and vaginal cytology, the swab obtained (cytobrush) should - artefact-free extraction of a typical lesion sufficient in size (diameter > 0.5 cm) be rolled onto a glass slide, not smeared. - immediate fixation (4% neutral buffered formaldehyde 10% formalin) All smears should generally be sent in air-dried, but unfixed and unstained. The most - preparation of an anamnesis including objective and clinical picture important point is to create a thin smear consisting of only one layer (monolayer). The - shipment in a suitable container (available from us free≙ of charge) most common reason for getting a limited quality up to not being able to assess at all - Immunohistochemistry can always be done after histopathology with the material are smears that are too thick. supplied.
18 19 2019/20 Pre-analytics 2019/20 Pre-analytics LABORATORY FOR CLINICAL DIAGNOSTICS 1.5 Polymerase chain reaction (PCR) swabs (without transport medium) should be supplied. To create DNA profiles and parentage reports in dogs and cats, we recommend to always send in a blood sample. PCR is a very sensitive and specific method for the direct detection of infectious For all genetic testing in horses, it is sufficient to supply about 20 hair roots from mane agents. Via PCR, gene sequences characteristic for the respective pathogen are or tail for DNA isolation. reproduced and detected – if necessary, even of germs which are no longer viable. EDTA blood is the most suitable sample material. It is absolutely essential to use EDTA The sample material that must be supplied for the PCR highly depends on the as anticoagulant. Lithium heparin or citrate are unsuitable as anticoagulants, as they pathogen to be detected and the present symptoms or the objective. Depending on may inhibit the subsequent PCR. In very rare cases, haemolysis induced by transport how the pathogen has spread in the body and its excretion, different sample materials or extreme stress during sample collection might lead to the situation that no result can are suitable. be obtained. However, the percentage of blood samples which cannot be evaluated is extremely low, being < 1%. At this stage of infection, pathogens causing viraemia, parasitaemia or bacteraemia can be detected directly in an EDTA blood sample (EB). Lithium heparin is less suitable as Buccal smears, often incorrectly called saliva samples, are very suitable sample materi- an anticoagulant, as it can inhibit the PCR. For blood samples or other liquid samples, als for genetic testing in dogs and cats, as long as the sampling procedure is performed an amount of at least 0.2 ml is required. correctly observing the following rules: 1. The animal should not have eaten anything for about 1 hour prior to the sample collec- In contrast to cultural bacteriological/mycological examinations, for PCR tests it is tion. It should be ensured that puppies and kittens have not been nursed for a minimum recommended to use sterile swabs without transport medium (“smear without of 2 hours, as otherwise maternal cells might skew the results. medium” = A, “swab without medium”, “dry swab”). If the concentration of the 2. When taking the sample, it should be scrubbed strongly at the inside of the cheek to pathogen is low, smears in a medium can lead to false negative results. For collecting make sure that enough cells of the oral mucosa and thus genetic material is attached to the sample, the swabs can be moistened with physiological saline solution. For PCR the swab. Genetic testing can only be conducted if enough genetic material adheres to tests, so-called cytobrushes (brush swabs) are also suitable, which can be shipped in the swab. Generally, saliva alone is not sufficient. However, there should not be any blood an uncoated sterile tube. on the swabs! 3. In order to prevent the growth of bacteria and mould, the swabs should be dried for For the detection of pathogens in faeces, a sample of approximately the size of a about 2 – 4 hours after collecting the sample. This is done best by keeping the test tubes hazelnut is needed. For some agents (e.g. coronavirus, Tritrichomonas foetus) we a little open for a while. recommend collecting faeces samples for 3 days, since these pathogens are excreted As there is considerably less cell material available from mucosal swabs compared to intermittently in the faeces. blood samples, it is not always possible to isolate enough DNA from buccal smears for Further sample materials, e.g. skin biopsies, organ material, urine, synovial fluid, liquor, a genetic test. This applies to about 5% of the submitted buccal swabs. We recommend bone marrow aspirates and lymph node aspirates, for PCR tests are best sent in sterile, sending two buccal samples per animal, so there is more material available for testing. uncoated test vessels. Fixation solutions such as formalin or the like can lead to DNA degradation, PCR inhibition and thus to false negative results. For horses, hair roots can be used to perform genetic examinations. To do so, about 20 Samples do not normally need to be sent cooled. Until it is dispatched, the sample pulled mane or tail hairs are needed. If samples are taken from various animals, hands material can be stored in the refrigerator at 2 – 8 °C. Repeated freezing/thawing should must be cleaned thoroughly after each sampling – even a single hair of a different animal absolutely be avoided. can skew the result. Hairs can, for instance, be shipped in little plastic bags or in envelopes. It is, however, Please note: Creating an antibiogram is not possible after doing a PCR test. absolutely necessary to make sure that the hairs are put in a closed envelope, separate from the submission form, when sent in.
There should not be any blood samples sent in for cattle from multiple births because of 1.6 Genetic testing a possible blood chimerism, but if the test allows it, hair roots, sperm or tissue samples can be used. One exception to this is the freemartin test, for which a blood sample is As sample material for the molecular genetic detection of hereditary diseases, for paren- mandatory. tage analysis as well as for the genetic determination of coat colours and blood types, If you wish to supply sample materials different from those listed above for performing EDTA whole blood samples (approx. 1 ml) are suitable. Alternatively, in dogs and genetic tests, please contact us before sending the samples. cats, buccal swabs, so-called cheek swabs, can be used. For each animal, 2 buccal
20 21 2019/20 Pre-analytics 2019/20 Pre-analytics LABORATORY FOR CLINICAL DIAGNOSTICS 1.7 Immune status (5) Citrate tube# (11) Shipping container for CB = Citrate blood: It can be swabs with/without medium EDTA blood samples must not be older than 48 hours! shipped in this tube (+ No. 8). CP = Citrate plasma: The sample should be centrifuged and the supernatant needs to be transferred 1.8 Sample material/Shipping material into a neutral tube (e.g. Eppendorf tube). It must then be marked accordingly as CP. (12) Urine tube Note regarding the sample materials listed in the test descriptions from Chapter 2 onwards: (suitable shipping container If the abbreviations are separated by a comma, you can choose the material which is (6) Salivette® see No. 8) easiest for you to collect from the given list. When collecting sample material for the for collecting saliva samples detection of a pathogen using PCR, you should preferably collect that material from the listed alternatives which is likely to have the highest concentration of pathogens. If the specifications are connected by one or more “+”, both or all the materials joined with “+” need to be provided for determining all the parameters of the selected testing (13) Container for histology block. (7) Blood smear (formalin tube with The materials required for the individual tests are also indicated on the submission Blood smears should always shipping container) forms, however, for lack of space, not always completely. be sent in air-dried, unfixed and unstained. For transportation, the Sample and shipping containers that are available for the collection and transport of depicted transport covers (shipping (14) Faeces tube the samples include: containers) are suitable. Before with shipping container transport, store at room temperature (25 °C) (may not be cooled).
(1) EDTA tube# (3) NaFB = Sodium fluoride blood (8) Shipping containers for EB = EDTA blood: It can be With NaFB samples, too, please pay blood tubes or urine tubes (15) Blood culture flask shipped in this tube (+ No. 8). attention to the labelling. EP = EDTA plasma: EDTA blood has to be centrifuged and the supernatant needs to be trans- ferred into a neutral tube (e.g. Eppendorf (4) S = Serum# (9) TM = Swab with tube). It must then be marked accordingly To collect serum, the coagulated transport medium # On special request, small test tubes as EP or labelled with the appropriate bar blood should be centrifuged at (orange: thin swab, (see each of the tubes shown on the left) code. 2000 g 30 minutes after being black: thick swab) are provided for collecting small amounts collected. The supernatant should of EDTA blood, EDTA plasma, heparin (2) Heparin tube# then be transferred into a neutral blood, heparin plasma, citrate blood, HB = Heparin blood: It can be tube or another serum tube (remove citrate plasma and serum, e.g. from small shipped in this tube (+ No. 8). beads before!) and marked accordingly mammals. If required, please order these HP = Heparin plasma: Heparin as serum or labelled with the appropriate small test tubes by e-mail or telephone blood has to be centrifuged and bar code. only. the supernatant needs to be transferred (10) A = Swab At your express request (information by into a neutral tube (e.g. Eppendorf tube). without transport medium, telephone or e-mail), we can also send It must then be marked accordingly as HP. (dry swab) you vacuum tubes for the examination of blood samples at Laboklin.
22 23 | 2019/20 Pre-analytics 2019/20 Pre-analytics |} LABORATORY FOR CLINICAL DIAGNOSTICS 1.9 Labelling Labeling of samples and shipping materials - The names of the animal and the owner should be clearly marked on the submission form and the sample. Alternatively, bar code labels can be used to unmistakably iden- Step 1 Step 2 1< GRG fq| G| Step 3 Step 4 V '1"$@W)02%3!#@ Test tube Additional X Y `a bcdecfg h`aiai pqidar i `id stiuvcd wxa`y`vv`g yaff p`a rdaf ffaf p`a ya`dax`f xaf x Sample into shipping bar code fdxsg y`a gayufd sciwvvaf Jane container onto the X Y `aiai pqidar h`afd sciiv`av` hax p`axa`dJane Sample c f h h a x wwa d` `d d `fAny Street xa 45 x v`f ` xs `i y `x t a ` shipping b vaiaf dxudehar svva fdxga iavtid cfh taxvsiiaf hsi f`d si`fafd City Anywhere, 12345 container X eahax sd`afd `id r`d a`far hcxgfg`gar pdx` gadxaffd haf axidaf sxuha fsd`afdbsr ag ffaf p`a wx x v`f`xs`ihc axyafhaf X Y `a sxuhataix`wdcfg h`afd sciiv`av` xaJane Sample Veterinarian x x`afd`axcfg cfh f`d hax cdursd`i`axcfg iuvvdBar code labels: af Any Street 45 Step 5 Step 6 p`a eii ey a` Y jhkxaf a`fai j`axai tavataf rCity Anywhere, 12345 iiaf ffaf p`a fsdxv` a`faf ya`daxaf r `d gva`ax bcr r ax axyafhaf cfh afdilxaafh hai sdax`svi taix`wdafi On one sheet, you will find the labels Completely fill Put sample and in submission submission X `dda axyafhaf p`a ta` raxaxaf j`axaf maya`vi fcx a`fa sxuhafcr rfor ax na`fa sxuhah 6 patients one below the other. For each patient, there are labels for form and, form and, if Y ullavxa`ao cfh hsec maya`vi a`faf fdxsg your laboratory journal, the submis- if necessary, necessary, sion form and to mark the sample sample list sample list, tubes/vessels to be sent. with sufficient cushioning p1'3!CD!'1"B There is a pre-printed label for serum material into the qi r sfe y`d`gs vi axidai iuvvda hax sxuha r`d and EDTA; for any other label, the cardboard box hax aix`wdcfg gfdxsgg scw hsi tavh material must be added in handwri- gcfhaffcr r ax sxuhag `r maya`v`gaf fdxsg ting. gavatd yaxhafi Please stick the bar code exactly Step 7 over the tube label so that the Please choose content is still visible through the the correct label Step 8 uncovered areas. and stick it on Pay attention to the box correct shipping ui Y`a sxuhai r`d haf aix`wdcfgaf Y j paxcr adi yaxhaf Exempt Animal Specimen information fgi scw h`a kxaf gavatdi i yaxhaf `rr ax fcx h`a xutafh xxaf r`d sxuhai axiaaf "%3!) $%# 43!1)0!D22# " v 24 25 wi sxuhai ufa aix`wdcfg ffaf ei i wx jclw ax udxxaf x x`f`vlxutaf yaxisfhdsia cfh fv`ai gafcded yaxhaf uhax wx ya`daxa Y jpaxcr hkxaf ma fs ahsxw zi fdxsg cfh xutaf `f h`a tvsca jda xs`iidarlav r`d javawuffcr r ax scw hsi tiafhaxwavh { wa xd`g d 2019/20 Pre-analytics 2019/20 Pre-analytics LABORATORY FOR CLINICAL DIAGNOSTICS 1.10 Packaging and transport 1.11 Reordering tests Please remember to pack your shipment according to EU regulations (European You can request additional tests for sample material that has already been sent in if Agreement concerning the International Carriage of Dangerous Goods by Road, ADR, • reordering is done within the sample storage period (see below) and International Air Transport Association, IATA): • the sample contains sufficient material Generally, transport containers that are transparent, break-proof and contain absorbent • the maximum sample age that may be indicated for the newly requested test is not material for leakage protection should be used and then packed, together with the exceeded (e.g. for morphology, FACS) submission form and cushioning material (not provided by the lab), in the transport (courier) box (min. dimension of 100x100x100 mm). Volume restriction: sample of 1000 Reordering can be done ml (applies to liquid samples) or a total weight of 4 kg (applies to illiquid samples). • by e-mail to [email protected] or to [email protected] LABOKLIN provides such protective outer packaging free of charge. • by telephone (+49 (0) 971 / 7202-0) via the switchboard or as part of our specialist counselling There are 2 possible categories of samples. The outer package needs to be tagged • by fax on +49 (0) 971 / 68546 according to the respective category with the labels shown in the illustration above. • by post An exempt animal specimen is a patient sample for which there is minimal likelihood that pathogens are present (e.g. blood, serum or formalin-fixed tissue samples). For reorders that shall be invoiced to the animal owner, please read the notes in Chapter 26. Classification must depend on professional judgement which is based on the anamnesis, the signs, the patient’s individual circumstances and local endemic Storage periods depend on the type of sample material and the purpose of the conditions. In case of doubt, it is recommended to ship as infectious substance of submission, i. e. the type of test that was originally requested. The periods specified Category B. below apply to samples tested in Bad Kissingen (as of July 2019). Infectious category B samples (swabs, urine, faeces etc.) must be marked as “Biological substance, Category B” and “UN 3373”; while the specification “UN 3373” Storage after clinical-chemical examinations, allergy tests, serological exami needs to be in a rhomb of at least 5 cm x 5 cm of size. The edge of the rhomb must be nations (antibody detection; antigen detection, except those from faeces): at least 2 mm wide and the letter height of both specifications must be at least 6 mm. • serum, heparin plasma, citrate plasma, EDTA plasma: 14 days • EDTA blood, heparin blood: 7 days Important: • urine: 7 days The sender is liable for the goods to be transported (i.e. sender is liable to recourse in • uroliths: 7 days (in most cases, however, the material is needed completely for case of damage/costs caused by samples that are not properly packed). analysis) If requirements are not met, there is a risk of your shipment being returned to you by the • punctures and liquor: 7 days courier company. (The same legislation applies to samples sent by courier service!) • blood smears: 7 days Please do not to leave any cannulae in the sample tubes! Storage after bacteriological, mycological and parasitological examination Do not seal the tubes! (detection by culture, all types of faecal analysis incl. antigen detection from If protective covers/transport containers are used, the lids will remain securely faeces), tests for maldigestion: closed. • faeces: 7 days • skin/hair, swabs, urine, milk: 14 days For shipment from a non-EU country, please contact LABOKLIN in advance. • punctures: 4 weeks Should you have any other questions, please do not hesitate to contact your local • isolated germs: 7 days LABOKLIN representative or contact us directly: [email protected] Storage after pathogen detection using PCR: • independent of the sample material (blood, liquor, urine, swabs, tissue, feather etc.): 3 weeks • extracted DNA/RNA: 1 year 26 27 2019/20 Pre-analytics 2019/20 Profiles - small animals LABORATORY FOR CLINICAL DIAGNOSTICS • Please note that extracted DNA/RNA is only suitable for reorders of further tests using PCR/genetic methods, but not for tests that require microbial growth. It is therefore 2 Profiles and Screenings not possible to request an additional resistance test if the sample had been sent in for PCR pathogen detection. Our profiles contain well-combined, complimentary parameters which offer a clear price advantage compared to individual requests. Results are usually transmitted on the day Storage after histopathological examination: the sample is received – on Saturdays, only partial findings might be transmitted. • wet material (tissue samples): 3 weeks • cytology – object slides/cytology – samples: 3 weeks For abbreviations and additional information concerning the test descriptions see page • paraffin blocks and sections: 5 years 11 and following. Storage after testing for hereditary diseases or coat colours/determination of breed, parentage: 2.1 Profiles/Screenings – Companion Animals extracted DNA: 5 years 2.1.1 Clinical Chemical Profiles Storage after sex determination in birds: extracted DNA: 1 year Adrenal Profile Material S 0.7 ml Parameter 17-OH-progesterone, androstenedione, oestradiol Note Profile for dog and ferret (Chapter 2.2.1) Anaemia Screening Basic Material S 1 m + EB 1 ml Parameter Complete blood count, reticulocytes, iron, protein, bilirubin total Anaemia Screening Cat Material S 1 ml + EB 1 ml Parameter Complete blood count, reticulocytes, iron, protein, bilirubin total, Coombs test + PCR: haemotropic mycoplasma Anaemia Screening Dog Material S 1 ml + EB 1 ml Parameter Complete blood count, reticulocytes, iron, protein, bilirubin total + PCR: babesia, Ehrlichia canis, Anaplasma phagocytophilum, haemotropic mycoplasma 28 29 2019/20 Profiles - small animals 2019/20 Profiles - small animals LABORATORY FOR CLINICAL DIAGNOSTICS BARF Profile Feline Profile Large Material S 3 ml + EB 1 ml (cooled!) Material S 1 ml + NaFB 1 ml Parameter ALT, creatine, protein, albumin, calcium, phosphate, copper, zinc, Parameter FIP, FeLV, FIV, Large Screening iodine, vitamin A, vitamin D3, vitamin E, T4, small blood count Note Complete blood count can be ordered in addition to this profile. Note See also Faecal Profile BARF (Chapter 2.1.6). In this case, please send 1 ml EB extra. See also Feed Ration Calculation in this chapter. Feline Profile Small Cardiac Screening Material S 1 ml Material S 1 ml (cooled!) Parameter FIP, FeLV, FIV, protein, albumin, albumin/globulin-ratio Parameter CK, LDH, α-HBDH, AST, calcium, magnesium, potassium, troponin FIP Screening (cat) Coagulation Status Material S 1 ml Material CP (1:10) 1 ml Parameter AST, bilirubin total, protein, electrophoresis, coronavirus antibodies Parameter PT, PTT, thrombin time Note Complete blood count can be ordered in addition to this profile. In this case, please send 1 ml EB extra. CSF Basis Test Material Cerebrospinal fluid FIV Monitoring (cat) Parameter Cell count, protein Material S 2 ml + EB 2 ml Parameter ALT, GLDH, AP, urea, creatinine, immune status (CD4/CD8; CSF Profile Small sample < 48h old), FIV PCR (quantitative)* Material Cerebrospinal fluid Note Complete blood count can be ordered in addition to this profile. Parameter Protein, IgA, CRP Geriatric Profile + SDMA Diabetes Monitoring Material S 1 ml + EB 1 ml Material S 1 ml + NaFB 1 ml Parameter LT, AST, GLDH, AP, bilirubin, urea, creatinine, fructosamine, protein, Parameter Glucose, fructosamine, creatinine, protein, β-HBS, lipase, ALT, AST, albumin, globulin, albumin/globulin-ratio, CK, potassium, calcium, sodium, potassium sodium, phosphate, iron, lipase, T4, SDMA Note Complete blood count can be ordered in addition to this profile. Note For male dogs, this profile can be also combined with CPSE. In this case, please send 1 ml EB extra. In this case, please send the serum cooled and centrifuged. DIC Profile Geriatric Profile + U-P/C Material EB 1 ml + CP 2 ml Material S 1 ml + EB 1 ml + urine 1 ml Parameter PT, PTT, thrombin time, D-dimer, complete blood count Parameter ALT, AST, GLDH, AP, bilirubin, protein, urea, creatinine, fructosamine, albumin, globulin, albumin/globulin ratio, CK, potassium, calcium, Feed Ration Calculation sodium, phosphate, iron, lipase, T4, urine protein/creatinine-ratio Note Compilation of an individual diet, please contact us. Note For male dogs, this profile can be also combined with CPSE. In this case, please send the serum cooled and centrifuged. 30 31 2019/20 Profiles - small animals 2019/20 Profiles - small animals LABORATORY FOR CLINICAL DIAGNOSTICS Juvenile Profile Note • It is recommended to send in more blood if possible, as flow cytometry requires up to 5 ml of sample volume if the total Material S 1 ml + EB 1 ml + faeces leukocyte count is low. Parameter ALT, AP, GLDH, calcium, protein, urea, creatinine, bile acids, phos- • For a complete evaluation in case of suspected leukaemia, phate, small blood count, endoparasites the leukaemia profile is always recommended and should be interprete in correlation with the clinical and the preliminary report. Kidney – Initial Screening • See also Leukaemia Classification (Leukaemia Differentiation, Material S 0.5 ml Chapter 7) Parameter Urea, creatinine Note Upon request, SDMA can be ordered in addition to this profile. Liver – Initial Screening Material S 0.5 ml Parameter ALT, GLDH, AP Kidney Screening Material S 1 ml Parameter Urea, creatinine, protein, albumin, sodium, potassium, calcium, Liver Screening 1 phosphate Material S 1 ml Parameter ALT, GLDH, AST (cat), AP, γ-GT (dog), bilirubin (I+II), protein, albumin, bile acids Leishmania Profile Large Material S 1 ml + EB 1 ml + urine 1 ml Parameter Leishmania ELISA, serum protein electrophoresis (incl. albumin, Liver Screening 2 globulins, albumin/globulin ratio), protein, creatinine, urea, ALT, AST, Material S 1 ml AP, GLDH, blood count Parameter AP, AST, ALT, protein, albumin, bile acids, manganese, copper Species Dog Note This profile can also be ordered in combination with the determination of the urine protein/creatinine-ratio (U-P/C) and microalbumin. Notation Recommended in case of shunt/cirrhosis/hepatitis. Leishmania Profile Small Muscular Screening Material S 1 ml + EB 1 ml Material S 1 ml Parameter Leishmania ELISA, serum protein electrophoresis (incl. albumin, Parameter CK, α-HBDH, AST, LDH, sodium, potassium, calcium, phosphate, globulins, albumin/globulin ratio), protein, creatinine, urea, ALT, magnesium, iron, sodium-potassium-ratio small blood count Muscular Screening Extended Leukaemia Profile Cat Material S 3 ml (cooled!) Material EB 3 ml + blood smear Parameter CK, LDH, AST, vitamin E, selenium, potassium, magnesium, Parameter Complete blood count, morphology, FeLV, FIV, clonality calcium, phosphate Leukaemia Profile Dog Pancreas Insufficiency (EPI) Profile Material EB 3 ml + blood smear Material S 1ml + NaFB 1 ml (fasted) Parameter Complete blood count, morphology, FACS (B- & T-cells, Parameter AST, ALT, albumin, sodium, potassium, chloride, calcium, glucose, stem cells (< 48h)), clonality TLI-assay, vitamin B12 32 33 2019/20 Profiles - small animals 2019/20 Profiles - small animals LABORATORY FOR CLINICAL DIAGNOSTICS Pancreatitis Profile Screening Small Material S 1 ml + NaFB 1 ml Material S 1 ml Parameter PLI, cholesterol, triglycerides, amylase, lipase, ALT, AST, protein, Parameter ALT, GLDH, lipase, urea, creatinine sodium, potassium, chloride, calcium, glucose Note Complete blood count can be ordered in addition to this profile. Note Complete blood count can be ordered in addition to this profile. In this case, please send 1 ml EB extra. In this case, please send 1 ml EB extra. Seizures Screening Cat Polydipsia Polyuria Screening Material S 1 ml + EB 1 ml + NaFB 1 ml Material S + EB + NaFB + urine each 1 ml Parameter Glucose, urea, calcium, phosphate, bile acids, small blood count, Parameter Glucose, fructosamine, ALT, GLDH, AP, sodium, potassium, calcium, toxoplasma antibodies protein, urea, creatinine, small blood count, urine status Seizures Screening Dog Pre-OP Screening Material S 1 ml + EB 1 ml + NaFB 1 ml Material S 1 ml + EB 1 ml + CP 1 ml Parameter Glucose, urea, calcium, phosphate, bile acids, small blood count, Parameter ALT, protein, urea, creatinine, small blood count, thrombocytes, Quick neospora antibodies, toxoplasma antibodies Pruritus Profile – Large (dog) T4 + TSH Material S 3.5 ml Material S 1 ml Parameter Seasonal and Perennial Panel, Food Allergens Basic and Extended, Parameter T4, TSH sarcoptes antibodies, flea saliva fT4 + TSH Pruritus Profile – Medium (cat, dog) Material S 1 ml Material S 2.5 m Parameter fT4, TSH Parameter Seasonal and Perennial Panel, Food Allergens Basic and Extended fT4 + TSH + Thyroglobulin Antibodies (dog) Pruritus Profile – Small (dog) Material S 1 ml Material S 2.5 ml Parameter fT4, TSH, thyroglobulin antibodies Parameter Allergy Screening Test (groups pollen, mites, moulds, fleas) and sarcoptes antibodies Thrombocyte Profiles Ø see Travel Profiles (Chapter 2.1.3) Screening Large Thyroid Profile Cat Anmerkung S 1 ml + NaFB 1 ml Material S 1.5 ml Parameter α-amylase, lipase, glucose, fructosamine, triglycerides, cholesterol, Parameter T4, fT4, T3, fT3, TSH, cholesterol bilirubin total, AP, GLDH, ALT, AST, CK, protein, albumin, globulins, γ-GT, urea, creatinine, phosphate, magnesium, calcium, potassium, Thyroid Profile Dog sodium, sodium-potassium-ratio, iron Material S 1.5 ml Note Complete blood count can be ordered in addition to this profile. Parameter T4, fT4, T3, fT3, TSH, thyroglobulin antibodies, T4 AB*, T3 AB* In this case, please send 1 ml EB extra. 34 35 2019/20 Profiles - small animals 2019/20 Profiles - small animals LABORATORY FOR CLINICAL DIAGNOSTICS TLI, B12, Folic Acid 2.1.3 Travel Profiles and Thrombocyte Profiles Material S 1 ml (fasted) Small Animals/Tick-borne Diseases Parameter TLI, vitamin B12, folic acid Anaemia Small (dog) Vitamin Profile Large Material EB Material EB 3 ml + S 3 ml (cooled!) Parameter PCR: Anaplasma phagocytophilum, babesia Parameter Vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin D (25 OH), vitamin E, folic acid Anaemia Vector-borne (dog) Note An additional order is possible up to a maximum of 1 day. Material EB Parameter PCR: babesia, Ehrlichia canis, Anaplasma phagocytophilum, Vitamin Profile Small haemotropic mycoplasma Material S, EP, HP 3 ml (cooled!) Parameter Vitamin A, vitamin D3 (25 OH), vitamin E, vitamin B12, folic acid Canine Travel Profile 1 Material S 1 ml Vomitus Profile Parameter Antibodies: leishmania, Ehrlichia canis, babesia Material S 0.5 ml + EB 1 ml Parameter AST, bile acids, protein, albumin, urea, sodium, potassium, chloride, Canine Travel Profile 2 large blood count Material S 1 ml + EB 1 ml Parameter Antibodies: leishmania, babesia, rickettsia, Ehrlichia canis PCR: hepatozoon, microfilaria (semiquantitative), Anaplasma platys 2.1.2 Serological Profiles Note Southern Spain, Balearic/Canary Islands, Portugal, Greece, Southern Italy, Croatia, Turkey, Albania, Southern Romania, Bulgaria, Serbia, FeLV, „FIP“, FIV Bosnia Material S, HP 0.4 ml Parameter Antibodies: FIP, FIV Canine Travel Profile 3 Antigen: FeLV Material EB 1 ml + S 1 ml Parameter Antibodies: leishmania, Ehrlichia canis, babesia, Anaplasma Herpes-/Calicivirus (cat) – Antibodies phagocytophilum Material S, HP 0.5 ml PCR: microfilaria (semiquantitative) Parameter FHV, FCV antibodies Note France, Northern Spain, Northern Italy, Slovenia Note Differentiation between infection and vaccination is only possible by means of testing serum pairs. Therefore, detection by PCR should Canine Travel Profile 4 be preferred. Material EB 1 ml + S 1 ml Parameter Antibodies: babesia, Anaplasma phagocytophilum, rickettsia Vaccination Control PCR: microfilaria (semiquantitative) Material S, HP 0.5 ml Note Poland, Czech Republic, Hungary, Ukraine, Slovakia, Northern Parameter Dog: distemper, parvovirus, adenovirus Romania, Russia Cat: calicivirus, herpesvirus, parvovirus (panleukopenia) 36 37 2019/20 Profiles - small animals 2019/20 Profiles - small animals LABORATORY FOR CLINICAL DIAGNOSTICS Canine Travel Profile Acute 2.1.4 Blood Donor Profiles – Small Animals Material EB 1 ml Parameter Small blood count Blood Donation Profile (cat) PCR: Anaplasma phagocytophilum, Anaplasma platys, babesia, Material EB + S + NaFB + urine 2 ml each hepatozoon, Ehrlichia canis, haemotropic mycoplasma) Parameter Blood group (serological) Complete blood count Feline Travel Profile Urea, creatinine, sodium, potassium, calcium, phosphate, bilirubin, ALT, AP, AST, GLDH, protein, albumin, glucose Material EB 1 ml + S 1 ml FIV antibodies, FeLV antigen Parameter Antibodies: leishmania, ehrlichia, Rickettsia felis Haemotropic mycoplasma (PCR) PCR: hepatozoon, microfilaria (semiquantitative) Urinalysis Leishmania Profiles Ø see Clinical Chemical Profiles (Chapter 2.1.1) Blood Donation Profile (dog) Thrombocyte Profile Large Material EB + S + NaFB + urine 2 ml each Parameter Blood group Material EB 1 ml Complete blood count Parameter Thrombocyte antibodies Urea, creatinine, sodium, potassium, calcium, phosphate, bilirubin, PCR: Anaplasma phygocytophilum, Ehrlichia canis, Anaplasma platys ALT, AP, GLDH, protein, albumin, glucose PCR: Anaplasma phagocytophilum, babesia Thrombocyte Profile Small Urinalysis Material EB 1 ml Parameter Thrombocyte antibodies, Blood Donation Profile/Follow Up (cat) PCR: Anaplasma phagocytophilum Material EB + S + NaFB + urine 2 ml each Parameter Complete blood count Tick I - PCR Urea, creatinine, sodium, potassium, calcium, phosphate, bilirubin, Material Tick ALT, AP, AST, GLDH, protein, albumin, glucose Parameter Borrelia, TBE-virus FIV antibodies, FeLV antigen Haemotropic mycoplasma (PCR) Urinalysis Tick II - PCR Material Tick Blood Donation Profile/Follow Up (dog) Parameter Anaplasma phagocytophilum, babesia, borrelia, TBE-virus Material EB + S + NaFB + urine 2 ml each Parameter Complete blood count Tick III - PCR Urea, creatinine, sodium, potassium, calcium, phosphate, bilirubin, Material Tick ALT, AP, GLDH, protein, albumin, glucose Parameter Anaplasma phagocytophilum, Anaplasma platys, babesia, borrelia, PCR: Anaplasma phagocytophilum, babesia Ehrlichia canis, Hepatozoon canis Urinalysis Vector-borne Diseases Material S, HP 1 ml Parameter Antibodies: borrelia, babesia, Anaplasma phagocytophilum 38 39 2019/20 Profiles - small animals 2019/20 Profiles - small animals LABORATORY FOR CLINICAL DIAGNOSTICS 2.1.5 PCR Profiles – Dog/Cat Reproduction Material Swab without medium (vagina, prepuce), abort material Anaemia Small (dog) Ø see Travel Profiles (Chapter 2.1.3) Parameter Dog: CHV, chlamydia, mycoplasma, Brucella canis Cat: FHV, chlamydia, Mycoplasma felis Anaemia Vector-borne (dog) Ø see Travel Profiles (Chapter 2.1.3) Respiratory I (cat) Diarrhoea Material Swab without medium (pharynx, nose, eye) Material Faeces Parameter FCV, FHV, chlamydia, Mycoplasma felis, Bordetella bronchiseptica Parameter Dog: coronavirus, parvovirus, circovirus, giardia, cryptosporidia Cat: coronavirus, Tritrichomonas foetus, giardia, parvovirus, cryptosporidia Respiratory II (cat) Material Swab without medium (pharynx, nose, eye) Diarrhoea, Human Pathogenic Causes Parameter FHV, FCV, chlamydia, Mycoplasma felis Material Faeces Parameter Salmonella, Yersinia enterocolitica, Campylobacter jejuni Respiratory III (cat) Material Swab without medium (pharynx, nose, eye) Eye (cat) Parameter FCV, FHV, chlamydia Material Swab without medium (eye) Parameter FHV, chlamydia, Mycoplasma felis Respiratory IV (cat) Material Swab without medium (pharynx, nose, eye) Eye (dog) Parameter FCV, FHV Material Swab without medium (eye) Parameter CHV, chlamydia, mycoplasma Respiratory Large (dog) Material Swab without medium (pharynx, nose, eye) Neurology (cat) Parameter CHV, CAV-2, CPiV, influenza A virus, distemper virus, Adenovirus, Bordetella bronchiseptica, mycoplasma Material CSF Parameter Coronavirus, Toxoplasma gondii, Bartonella henselae, bornavirus Respiratory Small (dog) Neurology (dog) Material Swab without medium (pharynx, nose, eye) Parameter CAV-2, CPiV, mycoplasma Material CSF Parameter Protein, IgA, CRP, AB: TBE Tick-borne Pathogen Detection Ø see Travel Profiles (Chapter 2.1.3) PCR: distemper virus, Neospora caninum, Toxoplasma gondii, Anaplasma phagocytophilum Neurology Small (dog) Material CSF Parameter Protein PCR: distemper virus, Neospora caninum, Toxoplasma gondii 40 41 2019/20 Profiles - small animals 2019/20 Profiles - small mammals, birds and reptiles LABORATORY FOR CLINICAL DIAGNOSTICS 2.1.6 Faecal Profiles – Small Animals Small Faecal Profile Parameter Bacteriology and mycology, facultative pathogenic bacteria, Culture Examination salmonella, clostridia The faeces tube should be ¾ full, if possible. An aerobe bacteriological and possibly Species Dog, cat mycological examination, including enrichment for salmonella and shigella, is perfor- med. Pathogen differentiation is done by MALDI-TOF. Unless otherwise stated, the test duration is 2 – 3 days. If required, serological pathogen differentiation (e.g. salmonella) Virological Faecal Examination and an antibiogram, which are subject to a charge, can be performed additionally. Combined Faecal Profile Virological Faecal Profile – EIA Parameter Bacteriology and mycology, facultative pathogenic bacteria, Parameter Parvovirus, rotavirus, coronavirus salmonella, clostridia, endoparasites, Giardia sp. antigen-EIA, Species Dog, cat Cryptosporidia antigen-EIA Species Dog, cat Faecal Profiles PCR Ø see Profiles „Diarrhoea“ (Chapter 2.1.5, PCR Profiles – Small Animals) Faecal Profile BARF Parameter Salmonella, yersinia, campylobacter, listeria, endoparasites Species Dog, cat 2.2 Profiles – Small Mammals, Birds and Reptiles Duration: 2 - 3 days (except Yersinia: 21 days) Note See also clinical chemical profiles, profile BARF (Chapter 2.1.1). 2.2.1 Clinical Chemical Profiles Small Mammals Faecal Profile Pathogenic Bacteria Adrenal Profile Parameter Salmonella, yersinia, campylobacter, enteropathogenic E. coli incl. virulence factors (STa, stx1, stx2, eae) Material S 0.7 ml Species: Dog Parameter 17-OH-progesterone, androstenedione, oestradiol Duration: 2 - 3 days (except Yersinia: 21 days) Note Profile for dog and ferret (Chapter 2.1.1) Large Faecal Profile Ferret Profile Parameter Bacteriology and mycology, facultative pathogenic bacteria, Material NaFB 0.5 ml + HP, S 0.5 ml salmonella, clostridia, Clostridium perfringens enterotoxin, Parameter Glucose, triglycerides, γ-GT, AST, CK, LDH, protein, albumin, Clostridium difficile toxin A and B, endoparasites globulin, urea, creatinine, calcium Species Dog, cat Rodent Profile Parasites Profile Cat Material S, HP 0.7 ml Parameter Endoparasites, giardia (EIA), PCR: Tritrichomonas foetus Parameter Urea, creatinine, protein, AST, ALT, GLDH, AP, γ-GT, CK, potassium, sodium, calcium, magnesium, bile acids, phosphate, fructosamine Puppy Faecal Profile Species Rabbit, guinea pig, rat, mouse, hamster Parameter Bacteriology and mycology, facultative pathogenic bacteria, Note Complete blood count can be ordered in addition to this profile. salmonella, clostridia, endoparasites incl. protozoa, parvovirus In this case, please send 0.5 ml EB or 0.5 ml HB extra or 1 ml HB in Species Dog, cat total. 42 43 2019/20 Profiles - small mammals, birds and reptiles 2019/20 Profiles - small mammals, birds and reptiles LABORATORY FOR CLINICAL DIAGNOSTICS Rodent Profile + T4 2.2.2 PCR Profiles – Small Mammals, Birds and Reptiles Material S, HP 0.7 ml Small Mammals Parameter Urea, creatinine, protein, AST, ALT, GLDH, AP, γ-GT, CK, potassium, sodium, calcium, magnesium, bile acids, phosphate, fructosamine, Rabbit: Respiratory Profile T4 Material Swab without medium Species Rabbit, guinea pig Parameter Bordetella bronchiseptica, toxigenic Pasteurella multocida, chlamydia Note Complete blood count can be ordered in addition to this profile. In this case, please send 0.5 ml EB or 0.5 ml HB extra or 1 ml HB in total. Birds Avian Profile I Small Profile (small mammals) Material EB, feather Material HP, S 0.4 ml Parameter Sex determination, PBFD Parameter Urea, creatinine, ALT, GLDH Species Rabbit, guinea pig, rat, mouse, hamster, ferret Avian Profile II Material EB, feather Parameter PBFD, polyomavirus Birds Avian Profile Avian Profile III Material S, HP 0.4 ml Material EB, feather Parameter LDH, AST, amylase, uric acid, bile acids, cholinesterase, CK, protein, Parameter Sex determination, PBFD, polyomavirus albumin, globulin, GLDH, sodium, potassium, calcium, phosphate Avian Profile IV Reptiles Material EB, feather Parameter Sex determination, PBFD, polyomavirus, herpesviruses Brumation Ø see PCR Profiles (Chapter 2.2.2) (e.g. Pacheco) Avian Profile V Reptile Profile (large) Material EB + swab without medium, feather + swab without medium Material S, HP 0.4 ml Parameter PBFD, polyomavirus, herpesviruses (e.g. Pacheco), chlamydia, Parameter AP, GLDH, ALT, AST, bile acids, CK, protein, albumin, urea, uric acid, bornavirus phosphate, calcium, potassium, sodium Avian Profile VI Reptile Profile (small) Material EB + swab without medium, feather + swab without medium Material S, HP 0.2 ml Parameter PBFD, polyomavirus, chlamydia Parameter Urea, protein, AST, AP, calcium, phosphate 44 45 2019/20 Profiles - small mammals, birds and reptiles 2019/20 Profiles - small mammals, birds and reptiles LABORATORY FOR CLINICAL DIAGNOSTICS Reptiles/Amphibians Respiratory/Neurology (pythons) Amphibian Profile Material Swab without medium + EB, tracheal lavage + EB Material Swab without medium, tissue Parameter Adenoviruses, arenaviruses, nidoviruses, paramyxoviruses/ Parameter Batrachochytrium dendrobatidis, Batrachochytrium ferlaviruses, reoviruses salamandrivorans, ranaviruses Respiratory Profile Large (tortoise) Brumation Check Large (tortoise) Material Swab without medium, nasal flush Material S, HP + HB + blood smear + swab without medium + faeces Parameter Herpesviruses, Mycoplasma agassizii, picornavirus Parameter Blood count, large reptile profile Antibodies: herpesvirus (TeHV-1 and TeHV-3) Respiratory Profile Small (tortoise, turtle) PCR: herpesviruses, mycoplasma Endoparasites Material Swab without medium, nasal flush Parameter Herpesviruses, mycoplasma Brumation Check Small (tortoise) Skin Profile (lizard) Material S, HP + swab without medium Parameter Large reptile profile Material Skin, swab without medium (skin) Antibodies: herpesvirus (TeHV-1 and TeHV-3) Parameter Adenoviruses, iridovirus, ranaviruses PCR: herpesviruses, mycoplasma 2.2.3 Faecal Profiles – Small Mammals, Birds and Reptiles Quarantine (boa/python) Material Swab without medium + EB, tracheal lavage + EB The faeces tube should be ¾ full, if possible. An aerobe bacteriological and possibly Parameter Adenoviruses, arenaviruses, paramyxoviruses/ferlaviruses, reoviruses mycological examination, including enrichment for salmonella and shigella, is performed. Pathogen differentiation is done by MALDI-TOF. Unless otherwise stated, Quarantine (colubrid/viper) the test duration is 2 – 3 days. If required, serological pathogen differentiation (e.g. salmonella) and an antibiogram, which are subject to a charge, can be performed Material Swab without medium, tracheal lavage additionally. Parameter Adenoviruses, paramyxoviruses/ferlaviruses, reoviruses Small Mammals Quarantine (lizard) Ferret Faecal Profile Material Swab without medium Parameter Adenoviruses, ranaviruses, reoviruses Material Faeces Parameter Bacteriology and mycology, salmonella, endoparasites, Giardia sp. antigen EIA Quarantine (tortoise) Material Swab without medium + HP, nasal flush + HP Rodent Faecal Profile + Endoparasites Parameter Herpesviruses, mycoplasma, picornavirus, ranaviruses, herpesvirus antibody (TeHV-1 and TeHV-3) Material Faeces Parameter Bacteriology and mycology, salmonella, endoparasites Species Rodent, rabbit Respiratory/Neurology (boa) Material Swab without medium + EB, tracheal lavage + EB Parameter Adenoviruses, arenaviruses, paramyxoviruses/ferlaviruses, reoviruses 46 47 2019/20 Profiles - small mammals, birds and reptiles / horse 2019/20 Profiles - horse LABORATORY FOR CLINICAL DIAGNOSTICS Birds EMS Profile (Equine Metabolic Syndrome) Avian Faecal Profile Material S 2 ml (cooled) + NaFB 2 ml Material Faeces Parameter Insulin, glucose, fructosamine, RISQI (reciprocal inverse square Parameter Bacteriology and mycology, salmonella of insulin), MIRG (modified insulin to glucose ratio), I/G-ratio (insulin:glucose) Avian Faecal Profile + Endoparasites Note Complete blood count can be ordered in addition to this profile. Material Faeces In this case, please send 1 ml EB extra. Parameter Bacteriology and mycology, salmonella, endoparasites MIRG is always calculated. If the value is not plausible, no result will be specified in the findings. Pigeon Faecal Profile Equine Cushing Profile Material Faeces Parameter Salmonella incl. enrichment, endoparasites (incl. coccidia) Material EP 2 ml (cooled) + S 2 ml (cooled) + NaFB 2 ml Parameter Insulin, ACTH, glucose, fructosamine, triglycerides, γ-GT, RISQI Reptiles (reciprocal inverse square of insulin), MIRG (modified insulin to glucose ratio), I/G-ratio (insulin:glucose) Reptile Faecal Profile Note Complete blood count can be ordered in addition to this profile. Material Faeces In this case, please send 1 ml EB extra. Parameter Bacteriology and mycology, salmonella Foal Profile Material S 1 ml 2.3 Profiles/Screenings – Horse Parameter Triglycerides, urea, creatinine, protein, electrophoresis, γ-GT, sodium, calcium, magnesium, phosphate 2.3.1 Clinical Chemical Profiles Note Complete blood count can be ordered in addition to this profile. In this case, please send 1 ml EB extra. Allergy Profile Respiratory Material S 1 ml Kidney – Initial Screening Parameter Seasonal, perennial panel Material S 0.5 ml Parameter Urea, creatinine Allergy Profile Skin Material S 3 ml Kidney Screening Parameter Seasonal, perennial, insect, food panel Material S 1 ml Parameter Urea, creatinine, protein, albumin, sodium, potassium, calcium, Blood Donation Profile phosphate Material EB + S + NaFB + CP + urine 2 ml each Parameter Complete blood count Liver – Initial Screening Fibrinogen Material S 0.5 ml Urea, creatinine, sodium, potassium, calcium, phosphate, bilirubin, Parameter AST, GLDH, γ-GT ALT, AP, AST, GLDH, protein, albumin, glucose Equine infectious anaemia virus (Coggins test), equine arteritis virus (VNT), babesia (c-ELISA) Urinalysis 48 49 2019/20 Profiles - horse 2019/20 Profiles - horse LABORATORY FOR CLINICAL DIAGNOSTICS Liver Screening Screening Large + SAA Material S 1 ml Material S, HP 1 ml + NaFB 1 ml Parameter AST, GLDH, AP, γ-GT, bilirubin (I+II), protein, albumin, globulins, bile Parameter AP, γ-GT, GLDH, bilirubin, triglycerides, cholesterol, glucose, AST, acids LDH, CK, protein, albumin, globulins, urea, creatinine, phosphate, calcium, magnesium, potassium, sodium, iron, copper, zinc, Mineral Profile II selenium, SAA Material S, HP 3 ml Note Complete blood count can be ordered in addition to this profile. Parameter Manganese, zinc, selenium, copper, sodium, potassium, calcium, In this case, please send 1 ml EB extra. magnesium, phosphate, chloride, iron Screening Small Muscular Screening Material S 1 ml Material S 1 ml Parameter GLDH, γ-GT, AST, LDH, CK, urea, creatinine, protein, triglycerides Parameter CK, α-HBDH, AST, LDH, sodium, potassium, calcium, phosphate, Note Complete blood count can be ordered in addition to this profile. magnesium, iron, sodium-potassium-ratio In this case, please send 1 ml EB extra. Muscular Screening Extended Senior Profile Material S 3 ml (cooled!) Material S 1 ml + NaFB 1 ml Parameter CK, LDH, α-HBDH, AST, vitamin E, selenium, potassium, Parameter Urea, creatinine, phosphate, calcium, bilirubin, γ-GT, GLDH, protein, magnesium, calcium, phosphate albumin, globulins, glucose, triglycerides, zinc, selenium Note Complete blood count can be ordered in addition to this profile. Performance Profile Horse In this case, please send 1 ml EB extra. Material S 1 ml + NaFB 1 ml Parameter AP, γ-GT, GLDH, bilirubin, triglycerides, cholesterol, glucose, lactate, Vitamin Profile Large AST, LDH, CK, protein, albumin, globulins, urea, creatinine, calcium, Material EB 3 ml + S 3 ml (cooled!) magnesium, potassium, sodium, iron, phosphate Parameter Vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin D (25 OH), vitamin E, folic acid Pruritus Profiles Ø see Allergy Profiles Note An additional order is possible up to a maximum of 1 day. Screening Large Vitamin Profile Small Material S, HP 1 ml + NaFB 1 ml Parameter AP, γ-GT, GLDH, bilirubin, triglycerides, cholesterol, glucose, AST, Material S, EP, HP 3 ml (cooled!) LDH, CK, protein, albumin, globulins, urea, creatinine, phosphate, Parameter Vitamin A, vitamin D3 (25 OH), vitamin E, vitamin B12, folic acid calcium, magnesium, potassium, sodium, iron, copper, zinc, selenium Note Complete blood count can be ordered in addition to this profile. In this case, please send 1 ml EB extra. 50 51 2019/20 Profiles - horse 2019/20 Profiles - horse LABORATORY FOR CLINICAL DIAGNOSTICS 2.3.2 PCR Profiles – Horse Respiratory Profile Material Swab without medium (nose), bronchoalveolar lavage Abortion Profile Parameter EHV1, EHV4, Influenza A virus Material Swab without medium, abort material Parameter EHV1, EHV4, EVA, leptospira Uveitis Profile Material Intraocular fluid 0.5 ml CEM Profile (mare) Parameter Leptospira AK, PCR: leptospira, EHV1 Material 2 swabs with medium of 2 locations: fossa clitoridis, sinus clitoridis Parameter Taylorella equigenitalis 2.3.3 Faecal Profiles – Horse Note Swabs must be analysed within 48 hours. The faeces tube should be ¾ full, if possible. An aerobe bacteriological and possibly CEM Profile (stallion) mycological examination, including enrichment for salmonella and shigella, is performed. Pathogen differentiation is done by MALDI-TOF. Unless otherwise stated, Material 3 swabs with medium of 3 locations: penile sheath, urethra, fossa the test duration is 2 – 3 days. If required, serological pathogen differentiation (e.g. glandis salmonella) and an antibiogram, which are subject to a charge, can be performed Parameter Taylorella equigenitalis additionally. Note Swabs must be analysed within 48 hours. Foal Faecal Profile Diarrhoea Pathogens (foal) Material Faeces Material Faeces Parameter Bacteriology and mycology, salmonella, gas producers, endoparasites Parameter Coronavirus, Lawsonia intracellularis, Rhodococcus equi incl. protozoa, rotavirus, Clostridium perfringens enterotoxin Eye Profile Large Faecal Profile Material Swab without medium (eye) Material Faeces Foals with respiratory symptoms: swab without medium Parameter Bacteriology and mycology, facultative pathogenic bacteria, (nose or pharynx), bronchoalveolar lavage salmonella, clostridia, Clostridium perfringens enterotoxin, Parameter EHV2+5 Clostridium difficile toxin A and B, endoparasites Parasites Profile Ø see Parasitology (Chapter 15) Respiratory Large Material Swab without medium (nose), bronchoalveolar lavage Small Faecal Profile Parameter EHV1, EHV4, EVA, Influenza A virus, Streptococcus equi equi/ zooepidemicus Material Faeces Parameter Bacteriology and mycology, facultative pathogenic bacteria, salmonella, clostridia Respiratory Profile (foal) Material Swab without medium (nose), bronchoalveolar lavage Parameter EHV1, EHV4, Influenza A virus, Rhodococcus equi 52 53 2019/20 Profiles - ruminants 2019/20 Profiles - ruminants LABORATORY FOR CLINICAL DIAGNOSTICS 2.4 Profiles/Screenings – Ruminants Mineral Profile Ruminants Material S, HP 1 ml 2.4.1 Clinical Chemical Profiles Parameter Calcium, sodium, phosphate, magnesium, selenium, zinc, copper Downer Cow Syndrome Mineral Profile II Material S, HP 1 ml Material S, HP 3 ml Parameter Calcium, phosphate, magnesium, AST, CK, urea, protein Parameter Manganese, zinc, selenium, copper, sodium, potassium, calcium, magnesium, phosphate, chloride, iron Fertility Profile (cattle) Material S, HP 1 ml Muscular Screening Parameter Calcium, phosphate, sodium, magnesium, AST, β-HBS, β-carotene Material S 1 ml Parameter CK, α-HBDH, AST, LDH, sodium, potassium, calcium, phosphate, Ketosis Profile magnesium, iron, sodium-potassium-ratio Material S, HP 1 ml Parameter GLDH, γ-GT, bilirubin, protein, β-HBS, NEFA, cholesterol, urea Performance Profile Ruminants Material EB 1 ml + S, HP 1 ml Kidney – Initial Screening Parameter Protein, urea, cholesterol, NEFA, GLDH, CK, γ-GT, bilirubin, calcium, phosphate, magnesium, β-HBS, β-carotene, glutathione peroxidase Material S 0.5 ml Parameter Urea, creatinine Note Complete blood count can be ordered in addition to this profile. Kidney Screening Screening Large Material S 1 ml Material S, HP 1 ml + NaFB 1 ml Parameter Urea, creatinine, protein, albumin, sodium, potassium, calcium, Parameter AP, γ-GT, GLDH, bilirubin, triglycerides, cholesterol, glucose, AST, phosphate LDH, CK, protein, albumin, globulins, urea, creatinine, phosphate, calcium, magnesium, potassium, sodium, iron, copper, zinc, selenium Llama/Alpaca Profile Note Complete blood count can be ordered in addition to this profile. In this case, please send 1 ml EB extra. Material S, HP 1 ml + NaFB 1 ml Parameter AP, γ-GT, GLDH, bilirubin, cholesterol, triglycerides, glucose, AST, LDH, CK, protein, albumin, globulin, urea, creatinine, calcium, Supply Profile Calf phosphate, sodium, magnesium, potassium, iron, copper, zinc, Material NaFB 1 ml + S, HP 1 ml (cooled!) selenium Parameter Glucose, protein, calcium, sodium, phosphate, magnesium, iron, Note Complete blood count can be ordered in addition to this profile. immunoglobulin G In this case, please send 1 ml EB extra. Supply Profile Calf + Vitamin E/Selenium Liver – Initial Screening Material NaFB 2 ml + S, HP 2 ml (cooled!) Material S 0.5 ml Parameter Glucose, protein, calcium, sodium, phosphate, magnesium, iron, Parameter AST, GLDH, γ-GT immunoglobulin G, vitamin E, selenium 54 55 2019/20 Profiles - ruminants 2019/20 Profiles - ruminants LABORATORY FOR CLINICAL DIAGNOSTICS Weak Calf Profile Bovine Respiratory Profile Material S, HP 1 ml Material Nasal flush, swab without medium + swab with medium Parameter Protein, calcium, phosphate, zinc, iron Parameter Bacteriology PCR: bovine respiratory syncytial virus (BRSV), bovine parainfluenza- Weak Calf Profile + Parasitology virus 3 (BPIV-3), Mycoplasma bovis Material S, HP 1 ml + faeces Parameter Protein, calcium, phosphate, zinc, iron, parasites incl. protozoa Bovine Respiratory Profile 2 Material Swab without medium (nose, pharynx), lavage (nasal flush, tracheal lavage, bronchoalveolar lavage 2.4.2 Serological Profiles – Ruminants Parameter Mannheimia haemolytica, Pasteurella multocida (toxin producing), Histophilus somni, Mycoplasma bovis Abortion Profile Camelids Mastitis PCR Profile* Material S, HP 2 ml Parameter Leptospira, chlamydia, Toxoplasma gondii Material Milk Parameter PCR test of 16 mastitis pathogens (incl. mycoplasma, yeasts, Prototheca spp. and β-lactamase gene (no antibiogram) Bovine Abortion Profile Material S, HP 3 ml Small Ruminant Abortion Profile Parameter Neospora caninum, Coxiella burnetii, leptospira, chlamydia, BVDV Material Abort material, swab without medium + swab with medium Parameter Bacteriology Bovine Respiratory Profile PCR: chlamydia, Coxiella burnetii Material S, HP 1 ml Parameter Bovine respiratoric syncytialvirus (BRSV), bovine parainfluenzavirus 3 (BPIV3), Mycoplasma bovis 2.4.4 Faecal Profiles – Ruminants Small Ruminant Abortion Profile Culture examination Material S, HP 3 ml The faeces tube should be ¾ full, if possible. An aerobe bacteriological and possibly Parameter Coxiella burnetii, chlamydia, leptospira, listeria mycological examination, including enrichment for salmonella and shigella, is perfor- med. Pathogen differentiation is done by MALDI-TOF. Unless otherwise stated, the test duration is 2 – 3 days. If required, serological pathogen differentiation (e.g. salmonella) 2.4.3 PCR Profiles – Ruminants and an antibiogram, which are subject to a charge, can be performed additionally. Abortion Profile Camelids Bovine Faecal Profile Material Abort material, swab without medium Material Faeces Parameter Leptospira, Toxoplasma gondii, chlamydia Parameter Bacteriology and mycology, salmonella, endoparasites, M. avium ssp. paratuberculosis (PCR) Bovine Abortion Profile Calf Faecal Profile – Large Material Abort material, swab without medium + swab with medium Parameter Bacteriology Material Faeces PCR: Neospora caninum, Coxiella burnetii, chlamydia, BVDV Parameter Bacteriology and mycology, salmonella, endoparasites, rotavirus, coronavirus 56 57 2019/20 Profiles - ruminants / pig 2019/20 Profiles - pig LABORATORY FOR CLINICAL DIAGNOSTICS Faecal Profile Camelids Muscular Screening Material Faeces Material S 1 ml Parameter Bacteriology and mycology, salmonella, C. perfringens enterotoxin, Parameter CK, α-HBDH, AST, LDH, sodium, potassium, calcium, phosphate, endoparasites, rotavirus, coronavirus magnesium, iron, sodium-potassium-ratio Porcine Profile Large Immunological Material S 1 ml Calf Faecal Profile (EIA) Parameter γ-GT, GLDH, bilirubin, ALT, AST, protein, CK, urea, creatinine, Material Faeces calcium, phosphate, sodium, magnesium, potassium, selenium, Parameter Rota-, coronavirus, E. coli K99, cryptosporidia zinc, copper Note Complete blood count can be ordered in addition to this profile. In this case, please send 1 ml EB extra. 2.5 Profiles/Screenings – Pig Porcine Profile Small 2.5.1 Clinical Chemical Profiles Material S 0.5 ml Parameter γ-GT, GLDH, bilirubin, urea, creatinine Kidney – Initial Screening Note Complete blood count can be ordered in addition to this profile. Material S 0.5 ml In this case, please send 1 ml EB extra. Parameter Urea, creatinine Kidney Screening 2.5.2 Serological Profiles – Pig Material S 1 ml Parameter Urea, creatinine, protein, albumin, sodium, potassium, calcium, Porcine Reproduction Profile phosphate Material S, HP 2 ml Parameter Leptospira, PRRSV*, chlamydia Liver – Initial Screening Material S 0.5 ml Porcine Respiratory Profile Parameter AST, GLDH, AP Material S, HP 3 ml Parameter APP, Mycoplasma hyopneumoniae, PRRSV* Liver Screening Material S 1 ml 2.5.3 PCR Profiles – Pig Parameter ALT, GLDH, AP, γ-GT, bilirubin (I+II), protein, albumin, bile acids Mineral Profile II Porcine Reproduction Profile Material S, HP 3 ml Material Abort material, swab without medium Parameter Manganese, zinc, selenium, copper, sodium, potassium, calcium, Parameter PPV, PRRSV, PCV-2, leptospira, chlamydia magnesium, phosphate, chloride, iron 58 59 2019/20 Profiles - pig / hygiene 2019/20 Haematology LABORATORY FOR CLINICAL DIAGNOSTICS Porcine Respiratory Profile 3 Haematology Material Nasal flush + swab with medium, swab without medium + swab with medium For abbreviations and additional information concerning the test descriptions see Parameter Bacteriology page 11 and following. PCR: Mycoplasma hyopneumoniae, APP*, PRRSV, Influenza A, Pasteurella multocida (toxin producing) 3.1 Blood Cells 2.5.4 Faecal Profiles – Pig small Blood Count The faeces tube should be ¾ full, if possible. An aerobe bacteriological and possibly Material EB, HB 1 ml mycological examination, including enrichment for salmonella and shigella, is perfor- Small mammals, birds: EB, HB 0.5 ml med. Pathogen differentiation is done by MALDI-TOF. Unless otherwise stated, the test Reptiles: HB 0.5 ml duration is 2 – 3 days. If required, serological pathogen differentiation (e.g. salmonella) Method Flow cytometry incl. peroxidase stain and an antibiogram, which are subject to a charge, can be performed additionally. Species Mammals, birds, reptiles (fish, amphibians on request) Duration 1 day Piglet Faecal Profile Note • For reliable results, the sample should not be older than 48 hours. Material Faeces • Small blood count includes erythrocytes, leucocytes, haemoglobin Parameter Bacteriology and mycology, salmonella, Clostridium perfringens and haematocrit; birds, reptiles: leucocytes, haemoglobin and enterotoxin, endoparasites, rotavirus, coronavirus haematocrit. Note If salmonella or E. coli are detected, they will be serologically typed as an additional service (subject to a charge). cytological Blood Smear Material Blood smear + EB, HB 1 ml Small mammals, birds: EB, HB 0.5 ml + blood smear Porcine Faecal Profile Reptiles: HB 0.5 ml + blood smear Material Faeces Method Microscopic Parameter Bacteriology and mycology, salmonella, Lawsonia intracellularis (PCR) Species Mammals Birds, reptiles, fish, amphibians on request Duration 1 day 2.6 Profiles – Hygiene Note • A differential incl. morphology of cells is useful in case of questionable leukaemia, anaemia or blood parasites. • An additional CBC is needed for full information. Hygiene Monitoring • Please add anamnesis or clinical issue. Hygiene monitoring includes testing of an autoclave OR a dry heat steriliser by means of bioindicators AND the examination of 3 surfaces (using contact plates) after disinfection. Bioindicators and contact plates need to be requested from the laboratory. Cultural examination of your samples will take 7 days. If you participate regularly (2x per year), you will get a certificate stating the successful annual monitoring of the disinfection performance of your autoclave/dry heat steriliser and the surface disinfection test. 60 61 2019/20 Haematology 2019/20 Haematology LABORATORY FOR CLINICAL DIAGNOSTICS Complete Blood Count Reticulocytes Material EB, HB 1 ml (+ blood smear) Material EB, HB 1 ml Small mammals, birds: EB, HB 0.5 ml (+blood smear) Small mammals: EB, HB 0.5 ml Reptiles: HB 0.5 ml (+ blood smear) Method Flow cytometry Method Mammals: flow cytometry incl. peroxidase stain; birds, reptiles: Species Dog, cat, small mammals, small ruminants, pig, others on request microscopic; flow cytometry incl. peroxidase stain Duration 1 day Species Mammals, birds, reptiles (fish, amphibians on request) Note • Reticulocytes are juvenile erythrocytes – determining their number Duration 1 day is necessary to be able to differentiate between regenerative and Note • For reliable results, the sample should not be older than 48 hours. non-regenerative anaemia. • Small blood count plus thrombocytes, evaluation of erythrocyte • For reliable results, the examination should be performed within 48 morphology and differential of leucocytes; dog/cat: additional de- hours after sampling. termination of reticulocytes and their haemoglobin concentration. • In dogs and cats, the haemoglobin concentration of reticulocytes • We advise to send a blood smear (air dried, not stained, unfixed) (CHr) is measured additionally. along with the EB sample/HB (reptiles) sample to provide maxi- mum information. Thrombocytes/Platelets Material EB, if applicable HB 1 ml Leucogram Method Flow cytometry Material EB, HB 1 ml (+ blood smear) Species Mammals, birds, reptiles (fish, amphibians on request) Small mammals, birds: EB, HB 0.5 ml (+ blood smear) Duration 1 day Reptiles: HB 0.5 ml (+ blood smear) Note • By far, most cases of haemostasis disorders in dogs are caused Method Mammals: flow cytometry incl. peroxidase stain by thrombocytopenia. Pre-surgery screens should thus include the Birds, reptiles: microscopic, flow cytometry incl. peroxidase stain determination of platelet numbers. Species Mammals, birds, reptiles (fish, amphibians on request) • Low counts are also seen in many cases of babesiosis and ana- Duration 1 day plasmosis. Note • Total leucocyte count is needed to interpret the differential blood • Platelet aggregates can lead to false low reports in many haema- count. tology machines. • Birds and reptiles: EB only in exceptional cases. • No microscopic platelet count. • Detection of platelet antibodies: see immunological tests MCHC, MCH, MCV (Chapter 7) and Thrombocyte Profiles (profiles/screenings-small animals, Chapter 2.1.3). Material EB 1 ml Method Flow cytometry incl. peroxidase stain Species Dog, cat, horse, ruminants, pig, others on request (not birds, reptiles) 3.2 Coagulation Duration 1 day Notation • The calculated erythrocyte indices help to differentiate between activated Clotting Time causes of anaemia. This test can be performed by veterinarians in their own practice. The appropriate tubes • Since the cell volume of erythrocytes varies with the ageing of the (ACT tubes) are subject to a fee and can be ordered from LABOKLIN. Please pay atten- blood, the indices have to be interpreted with caution in shipped tion to the detailed instructions for use included in the delivery. samples. Coagulation Status Ø see Chapter 2.1.1 62 63 2019/20 Haematology 2019/20 Haematology LABORATORY FOR CLINICAL DIAGNOSTICS D-Dimer Prothrombin Time (PT) Material CP (1:10) 0.5 ml Material CP (1:10) 1 ml Method Turbidimetry Method Ball coagulometer Species Dog, cat, horse Species Dog, cat, cattle, horse, others on request Duration 1 day Duration 1 day Note D-dimers are generated by lysis of cross-linked fibrin. D-dimers are, Note This test comprises the coagulation factors of the extrinsic system. for example, detectable if there are internal bleedings as well as in It has to be taken into account, however, that levels may be normal surgical interventions and neoplasia. Particularly high amounts of in chronic coagulopathy. Thromboplastin time is used as diagnostic D-dimers are generated in case of thromboembolism and dissemi- aid in suspected poisoning with vitamin K antagonists (coumarin nated intravascular coagulation (DIC). In diagnostic work, D-dimers and warfarin derivatives) and for therapy monitoring while vitamin K are mostly used for DIC. is administered. DIC Profile Ø see Chapter 2.1.1 partial Thromboplastin Time (PTT) Material CP (1:10) 1ml Factor VIII Method Ball coagulometer Material CP (1:10) 1 ml Species Dog, cat, cattle, horse, others on request Method Ball coagulometer Duration 1 day Species Dog, cat, horse Note • PTT is used to monitor coagulation factors of the intrinsic system Duration 1 day and can be used as global test to identify coagulopathies. Note • Factor VIII deficiency is the most common single factor deficiency • An isolated prolongation of PTT without changes in thromboplastin and the cause of haemophilia A. may indicate a factor deficiency (factor VIII, IX, XI and XII). Haemo- • The determination of single factors is only useful if there are chan- philia A and B can be identified by determining the concentration ges in partial thromboplastin time. of single factors (VIII and IX). Factor IX Thrombin Time Material CP (1:10) 1 ml Material CP (1:10) 1 ml Method Ball coagulometer Method Ball coagulometer Species Dog, cat, horse Species Dog, cat, cattle, horse, others on request Duration 1 day Duration 1 day Note • A lowered concentration of factor IX is the cause of haemophilia B. Note • This test covers the third phase of coagulation, the change from • The determination of single factors is only useful if there are fibrinogen to fibrin. changes in partial thromboplastin time. • It is recommended for monitoring the treatment with heparin or streptokinase as well as in cases of suspected disseminated intra- Fibrinogen vascular coagulopathy or intoxication with vitamin K antagonists. Temporarily lowered concentrations of fibrinogen are seen after Material CP (1:10) 1 ml intensive surgery as well as in cases of disseminated intravascular Method Photometry coagulation (DIC). Species Dog, cat, cattle, horse • In case of suspected DIC, the DIC profile may be used to confirm Duration 1 day or disprove the diagnosis (see Chapter 2.1.1). Note • Determination is recommended in case of possible disseminated intravascular coagulopathy or hypofibrinogenemia. Thrombocyte Antibodies Ø see Chapter 7 • As fibrinogen is an acute phase protein, the concentration will rise Thrombocyte Profiles Ø see Chapter 2.1.3 in case of acute inflammation. 64 65 2019/20 Haematology 2019/20 Haematology LABORATORY FOR CLINICAL DIAGNOSTICS Thromboelastography identify the recessive b allele which is associated with the B seroty- pe. Cats with two copies of the b allele have blood type B. Cats with Material CB 2 ml blood type A can carry the genotype AA or Ab. To clarify the genetic Method Thromboelastography basis with blood type A or AB, the genetic test is recommended. Species Dog, cat, cattle, horse (See also Chapter 20.3.1) Duration 1 day Note • The citrate blood sample should not be older than 48 hours. Serological Blood Grouping • Global test to determine coagulation disorders, including DIC and thrombocytopathies. Material EB 1 ml • If DIC is suspected, the DIC profile is also available for diagnostic Method Agglutination assessment (Chapter 2.1.1). Species Dog, cat Duration 1 – 2 days, it is possible to send out rapid blood typing tests for use in the practice Von Willebrand Antigen Note Dog: Material CP (1:10) 1 ml (cooled or frozen) • DEA 1,1 positive/negative Method Immunoturbidimetry • Prior to blood transfusions, it is advisable to test donor and Species Dog recipient animals for blood group compatibility. Duration 1 day Cat: Note • Determination of the von Willebrand antigen is used for further • A, B, AB evaluation of coagulation disorders. • Do not use umbilical cord blood. • The von Willebrand disease (vWD) has been described in many • In cat breeds, the blood groups of the parent animals should be dog breeds; only genetic testing can detect whether the disease is determined before mating in order to avoid neonatal iso-immune carried. haemolytic anaemia. Genetic testing in A animals is indicated to detect carriers of the recessive B gene. 3.3 Blood Grouping Genetic Blood Grouping (cat) Material EB 1 ml, buccal swab Method TaqMan SNP assay Species Cat Race All (except European Shorthair) Duration 3 – 5 days Note The AB system is the major blood group system in domestic cats. The most common blood types are A and B. Cats with blood type B usually have high anti-A antibody titres and cats with blood type A usually have low anti-B antibody titres. Cats with the rare AB blood type (also called blood type C) do not have anti-A or anti-B antibodies and are thus universal receivers in case of blood trans fusions. Phenotype AB is not caused by a co-dominance of A and B. Genetic blood grouping in cats allows for genetic differentiation of the serologically determined blood group before breeding. Blood type A is dominant to B. With the genetic test, it is possible to 66 67 2019/20 Clinical Chemistry 2019/20 Clinical Chemistry LABORATORY FOR CLINICAL DIAGNOSTICS • Young animals: physiological concentration up to 2.5-fold. 4 Clinical Chemistry • Dog: Diagnosis of corticosteroid-induced AP is possible by determining the heat-stable isofraction. For abbreviations and additional information concerning the test descriptions see • Cat: only increase is specific. page 11 and following. • Cattle: The AP level ante partum allows to assess the risk of parturient paresis. 4.1 Enzymes AP (heat-stable 65°C) (heat-stable Alkaline Phosphatase) ALT (GPT) Material S 0.5 ml (Alanine Aminotransferase) Method Photometry Material S, EP, HP 0.5 ml Species Dog Method Photometry Duration 1 day Species Dog, cat, rabbit, guinea pig, rat, mouse, ferret, birds, reptiles, cattle, Note • The heat-stable isoenzyme of AP is induced by endogenous steroid goat, sheep, pig hormones or by cortisone therapy and can be used to diagnose Duration 1 day overtreatment with steroids. Notation • In contrast to horses, cattle and pigs, this parameter is liver- • Only useful in combination with the determination of the “total specific in dogs and cats. ALT is found only in the cytoplasm, AP” if it is elevated. With the sum of all isoenzymes of AP (“total therefore even minor cell damage may cause elevated levels. AP”) and the heat-stable AP, the residual activity of AP can be Isolated elevations also occur in case of portosystemic shunt. determined in percentage. • Cat: In Oriental breeds, the reference range is higher than in other cat breeds. AST (GOT) Aspartate Aminotransferase (Glutamate Oxaloacetate Transaminase) AP (Alkaline Phosphatase) Material S, EP, HP 0.5 ml Material S 0.5 ml Method Photometry Method Photometry Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, horse, Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, horse, ruminants, pig ruminants, pig Duration 1 day Duration 1 day Note • Elevated levels can be caused by diseases of various Note • The enzyme is found in almost all organs. AP is diagnostically parenchymatous organs but also by muscle damage. If the latter, especially significant in diseases of the skeletal and the it cannot be distinguished between damage of skeletal muscles hepatobiliary system. In dogs, there is also steroid-induced and cardiac muscles. A simultaneous increase of CK indicates a AP, which particularly plays a role in the diagnosis of myogenic origin. hyperadrenocorticism (Cushing‘s disease). • Cat: sensitive marker of hepatopathies; to differentiate muscle • In the context of bone diseases, high levels are present in case of damage, CK levels should be determined additionally. ostitis deformans, which allows to differentiate from osteoporosis. • Horse: indicates lesions of the skeletal muscles (in combination In bone tumours, increases in activity are measured whose with other parameters, for example LDH, CK) or the liver. extent correlates to the osteoblast activity (very high levels in osteosarcoma, hardly any increases in benign tumours). Rachitis and osteomalacia have elevated levels with decreased calcium levels. • Increased levels may indicate cholestasis. 68 69 2019/20 Clinical Chemistry 2019/20 Clinical Chemistry LABORATORY FOR CLINICAL DIAGNOSTICS Cholinesterase Glutathione Peroxidase (GPx) Material S, EP, HP 0.5 ml Material EB, HB 0.5 ml Method Photometry Method Photometry Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, horse, others Species Horse, ruminants, pig on request Duration 1 day (except for Saturdays) Duration 1 day Note • GPx can only be determined in unclotted whole blood samples. Note • In case of an intoxication with organophosphoric acid esters, the • GPx is an antioxidant. enzyme is blocked and its activity in the blood plasma decreases. • As GPx is a selenium-containing enzyme, it indicates an adequate • Birds: liver-specific, increased in case of chronic processes supply of selenium to the animal. Since the impact of supplied selenium can only be determined after 2 – 3 weeks, GPx cannot CK (Creatine Kinase) indicate an acute oversupply. • The GPx value is linked to the haemoglobin. Material S, EP, HP 0.5 ml Method Photometry Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, horse, LDH (Lactate Dehydrogenase) ruminants, pig Material S, EP, HP 0.5 ml Duration 1 day Method Photometry Note The highest enzyme activity by far is found in the skeletal muscles, Species Dog, cat, rabbit, guinea pig, rat, ferret, horse, ruminants, pig followed by brain tissue and cardiac muscles. All conditions that Duration 1 day cause destruction of muscle cell membranes result in elevated Note • LDH is composed of 5 isoenzymes. It is found in many serum concentrations (physiologic: training; pathologic: e.g. myo organs, mainly in the liver, cardiac and skeletal muscles. High pathies, traumas caused by i.m. injections). Preanalytically, haemo concentrations of LDH are also present in red blood cells, so that lysis also leads to elevated levels. Brain tissue damage does not even slight haemolysis in serum or plasma may cause falsely cause increased serum levels because of the blood-brain barrier. elevated values. • Elevations occur in case of myopathies, cardiomyopathies and GLDH (Glutamate Dehydrogenase) liver diseases. • The ratio of α-HBDH to LDH can indicate problems of the cardiac Material S, EP, HP 0.5 ml or skeletal muscles. Method Photometry Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, horse, ruminants, pig Lipase Duration 1 day Material S (EP, HP) 0.5 ml Note The enzyme is liver-specific and located in the mitochondrion. Method Photometry (DGGR) Thus, elevations are indicative of massive destruction of liver cells Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, others on (mitochondrial enzymes) and necrobiotic processes, especially in request the centrilobular area. If levels of GLDH are increased, but at the Duration 1 day same time ALT levels are changed only slightly, it indicates chronic Note • Measurement mainly covers the activity of pancreatic lipase, but inflammation of the liver. also the lipase activity in other tissues (stomach, small intestine). A • Dog: Single values have no diagnostic significance. Slight threefold increase in value indicates acute pancreatitis. elevation of GLDH and stronger elevation of transaminases • In serum, lipase activity remains elevated much longer than indicate acute disease of the liver. Opposite enzyme activity amylase activity. Therefore, if pancreatitis is suspected, both indicates chronic processes. enzymes should always be determined. What applies to both • Cat: In Oriental breeds, the reference range is higher than in other enzymes is the fact that the enzyme concentration is not cat breeds. proportional to the severity of illness. Especially in cats, unaffected • Cattle: Levels depend on stage of lactation. levels of lipase can be seen despite pancreatitis. 70 71 2019/20 Clinical Chemistry 2019/20 Clinical Chemistry LABORATORY FOR CLINICAL DIAGNOSTICS • In order to confirm the diagnosis of pancreatitis, pancreatic lipase α-HBDH (α-Hydroxybutyrate Dehydrogenase) immunoreactivity (PLI) should be determined. • Lipase levels are also elevated after administration of Material S, EP, HP 0.5 ml glucocorticoids. Method Photometry Species Dog, cat, horse, ruminants, pig Duration 1 day PLI (Pancreatic Lipase Immunoreactivity) Note • This isoenzyme of LDH is found in many kinds of tissue, especially Material S 0.5 ml in cardiac and skeletal muscles and in the liver; the activity of Method ELISA α-HBDH varies depending on the species. Species Dog, cat • LDH/HBDH ratio can indicate possible damage of the cardiac Duration 1 day muscle (positive if ratio exceeds 1:2). Determination of c-Troponin I Note • Detection of specific pancreatic lipase in suspected pancreatitis. concentration has replaced the analysis of α-HBDH in case of this The determination of pancreatic lipase in the serum of dogs and indication, though. cats is considered to be the most sensitive non-invasive marker for • Proportional or slight elevations of the enzyme point to other the diagnosis of pancreatitis. As part of an inflammatory reaction, causes (e.g. liver damage, damage of skeletal muscles, the pancreatic acinar cells are destroyed leading to an increase in haemolysis and others). In this case, CK and AST levels should be the pancreatic lipase concentration in the serum. considered. • If possible, 12 hours of fasting is recommended. γ-GT (γ-Glutamyl Transferase) TLI-Test (Trypsin-like Immunoreactivity) Material S, EP, HP 0.5 ml Material S 0.5 ml Method Photometry Method CLIA, ELISA Species Dog, cat, rabbit, guinea pig, rat, ferret, horse, ruminants, pig Species Dog, cat Duration 1 day Duration Dog: 1 day, Cat: 2 – 3 days Note • Although this membrane-bound enzyme is not liver-specific, Note • Most sensitive test for the de- elevations occur almost exclusively in diseases of the liver and bile tection of exocrine pancreatic ducts. insufficiency. • Horse: Elevated concentrations are indicative of cholestasis. • Elevated levels can indicate Increased levels may also be seen in other diseases with liver pancreatitis in dogs and cats. involvement, such as colic, enteritis and the like. • Dogs and cats must fast for • Cattle: The γ-GT level strongly correlates with the degree of 12 hours prior to sampling. hepatic fatty degeneration and the degree of swelling of the liver and the edge of the liver. Decreased levels of this enzyme indicate insufficient intake of colostrum in calves. α-Amylase Material S, EP, HP 0.5 ml Method Photometry Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, horse, cattle, pig Duration 1 day Note • The enzyme is elevated in the acute phase of pancreatitis for 3 – 5 days. Slight elevations also occur due to diseases of other organs and in case of renal dysfunction. As the enzyme is also produced in the liver and small intestine, it is not pancreas-specific. Therefore, its suitability for diagnosing pancreatitis is limited. • In order to confirm the diagnosis of pancreatitis, determination of pancreatic lipase immunoreactivity (PLI) is recommended. 72 73 2019/20 Clinical Chemistry 2019/20 Clinical Chemistry LABORATORY FOR CLINICAL DIAGNOSTICS 4.2 Substrates horses, visible icterus correlates with concentrations of 17 μmol/l and more. • Prehepatic icterus: Excessive haemoglobin concentration due to Albumin erythrocyte destruction causes increased levels of indirect bili Material S, EP, HP 0.5 ml rubin. Method Photometry • Intrahepatic icterus: Damage of liver cells causes increase in both Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, horse, direct and indirect bilirubin. ruminants, pig • Posthepatic icterus (rare): Increase in direct bilirubin caused by Duration 1 day retention of bile. Note • Absolute hypoalbuminaemia occurs in cases of nephropathy • Cattle: Total bilirubin has a strong negative correlation with blood and enteropathy. In cattle it is found particularly in cases of glucose levels and hence is a sensitive indicator for imbalances hepatic diseases, reduced feed intake and inflammation. Relative in the composition of food rations. Strong increase occurs due to hypoalbuminaemia is detected if shifts in total protein occur in microhaemolysis as part of septicaemia, e.g. in case of mastitis, favour of globulins. endometritis or salmonellosis, and is prognostically unfavourable. • Albumin is also a negative acute phase protein. • Increased levels only occur in the form of relative Bilirubin II (direct) hyperalbuminaemia in case of dehydration. Material S, EP, HP 0.5 ml Method Photometry Bile Acids Species Dog, cat, rat, ferret, horse, others on request Material S 0.5 ml Duration 1 day Method Photometry Note In the liver cells, bilirubin II is formed from bilirubin I by glucuroni Species Dog, cat, rabbit, guinea pig, birds, reptiles, horse, cattle dation. Determination is only useful in case of elevated levels of total Duration 1 day bilirubin. Measurement can be strongly affected by lipidaemia. Note • Serum bile acid concentration correlates with liver function. In contrast to the determination of ammonia, which is very Cholesterol susceptible to faults and needs to be performed immediately after Material S, EP, HP 0.5 ml sampling, this parameter is very stable. Method Photometry • Elevation of bile acids can also indicate the existence of a Species Dog, cat, rabbit, guinea pig, rat, ferret, horse, ruminants, pig portosystemic shunt. Duration 1 day • Single determinations can lead to false normal results, therefore, the performance of bile acid stimulation tests is preferable – Note • Cholesterol is formed mainly in the liver and in the mucosa of the except for horses. small intestine and serves as starting material for many com- • Please note that animals must fast for 12 hours before sampling pounds which are synthesised in the liver (e.g. bile acids and (except horses). steroid compounds). • In cattle, cholesterol levels correlate with feed intake and milk yield. Bilirubin (total) • Please note: Fasting is needed 12 hours prior to sampling in Material S, EP, HP 0.5 ml order to obtain relevant results! Method Photometry Species Dog, cat, rabbit, guinea pig, rat, ferret, horse, ruminants, pig Cholesterol: HDL (High Density Lipoproteins) Duration 1 day Material S, EP, HP 0.5 ml Note Bilirubin is formed in the liver as part of the decomposition of Method Photometry haemoglobin and other cytochromes. Its glucuronidation is Species All intrahepatocellular and it is excreted through the intestines. Except in Duration 2 days 74 75 2019/20 Clinical Chemistry 2019/20 Clinical Chemistry LABORATORY FOR CLINICAL DIAGNOSTICS Cholesterol: LDL (Low Density Lipoproteins) Fructosamines Material S, EP, HP 0.5 ml Material S, EP, HP 1 ml Method Photometry Method Photometry Species All Species Dog, cat, rabbit, guinea pig, rat, ferret, horse Duration 2 days Duration 1 day Note • This glycoprotein reflects the average blood glucose level of the Creatinine past 3 weeks, determination therefore serves the diagnosis and Material S, EP, HP 0.5 ml the long-term monitoring of patients with diabetes mellitus. A Method Photometry distinction between spontaneous, stress-induced hyperglycaemia Species Dog, cat, rabbit, guinea pig, rat, ferret, horse, ruminants, pig and persistent hyperglycaemia due to diabetes mellitus is possible Duration 1 day as well. • Horse: often increased in case of EMS. Note • Creatinine is the most specific parameter for kidney function. However, due to the reserve capacity of the kidneys, elevated levels only occur if the kidney damage exceeds 70%. Lipaemia Glucose and haemolysis can cause false elevation of values. In Material NaFB 1 ml, CSF, H well-muscled or trained dogs, creatinine may be increased Method Photometry physiologically, without there being a renal dysfunction. Species Dog, cat, rabbit, guinea pig, rat, mouse, ferret, birds, reptiles, horse, • Protein/creatinine ratio in the urine (midstream urine or urine from ruminants, pig cystocentesis) and SDMA serve for an early detection of renal Duration 1 day dysfunction. Note • Increased blood glucose levels occur in case of diabetes mellitus • Cattle: An increase in creatinine is an important indicator for but also due to brain diseases, pancreatitis and Cushing’s insufficient feed intake/body weight loss. disease. Levels particularly increase physiologically due to stress and after administration of glucocorticoids. Cystatin C • Diseases of the liver, Addison’s disease and insulinoma cause Material S 0.5 ml hypoglycaemia. Method Photometry • Drugs which can lead to hypoglycaemia are (among others): Species Dog, cat antihistamines, beta blockers, anabolic steroids Duration 1 day • Dog: Starving young dogs of toy breeds tend to develop life- threatening hypoglycaemia in stress situations. Note • Parameter for early renal diagnosis in dogs and cats. Most • Horse: The determination of glucose levels is a necessary part of the nucleated cells produce cystatin C at a constant rate; of the diagnosis of Equine Metabolic Syndrome. For further the production also seems to remain unaffected in cases of information please see „Insulin“. inflammation or other pathological conditions. Levels can be • In cattle, hypoglycaemia indicates ketosis due to energy deficit; increased in animals suffering from manifest hyperthyroidism or hyperglycaemia is caused by stress and endotoxaemia. after administration of glucocorticoids. • Cystatin C is filtered in the kidney, probably also tubularly HDL Ø see Cholesterol secreted, 99% reabsorbed in the proximal tubule and then degraded. Lactate Material NaFB 0.5 ml Method Photometry Species Dog, cat, horse, cattle, pig Duration 1 day 76 77 2019/20 Clinical Chemistry 2019/20 Clinical Chemistry LABORATORY FOR CLINICAL DIAGNOSTICS Note • Anaerobic degradation of glucose leads to the formation of SDMA (Symmetric Dimethylarginine) lactate. Elevated lactate concentration may be caused by increased formation of lactate because of increased intake of Material S, HP 0.5 ml glucose or increased glycogenolysis (e.g. in case of diabetes Method ELISA mellitus), impaired metabolisation (hypovolemic, cardiovascular Species Dog, cat or neurogenic shock) and enhanced formation due to oxygen Duration 1 – 2 days deficiency in the tissue (training condition, stress due to blood Note SDMA is formed during protein degradation, is excreted exclusively sampling, increase of lactate in immature neonates). via the kidneys and is helpful for the early detection of renal • Cattle: Elevation occurs in case of ruminal acidosis, circulatory dysfunction, even in the creatinine-blind area. In cats, a significant disorders, severe pneumonia. inverse correlation between the glomerular filtration rate and SDMA has been described. Determination of SDMA is recommended if LDL Ø see Cholesterol an early stage of renal dysfunction is suspected, e.g. because of beginning polyuria/polydipsia. Increased levels signify a reduction in NEFA (Non-Esterised Free Fatty Acids) renal function by 40%. Material S, HP 0.5 ml Method Photometry Taurine* Species Ruminants, pig Material S 1 ml Duration 1 day Method LCMS/mass spectroscopy Note The degradation of adipose tissue releases NEFA (non-esterised Species Dog, cat free fatty acids). They are a quick and sensitive marker of nutritional Duration 7 days deficiency or of reduced feed intake in case of stress situations or Note Chronic taurine deficiency causes dilatative cardiomyopathy in cats. disease, and serve to estimate e.g. the fat mobilisation in metabolic In general, commercial diets contain sufficient amounts of taurine. states. Taurine deficiency may be caused by chronic malabsorption or by homemade rations. Protein Material S, EP, HP, CSF 0.5 ml, H Triglycerides Method Photometry Material S, EP, HP 0.5 ml Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, horse, Method Photometry ruminants, pig Species Dog, cat, rabbit, guinea pig, rat, ferret, reptiles, horse, ruminants, pig Duration 1 day Duration 1 day Note • The main tasks of plasma proteins are water retention, transport, Note • Intake with food and synthesis in the liver. coagulation and defence. • Horse: Hyperlipaemia in ponies, metabolic syndrome and • Absolute hyperproteinaemia usually occurs due to chronic Cushing’s disease. infection, relative rises are common in case of fluid loss. Absolute • Cattle: Lipid mobilisation syndrome hypoproteinaemia occurs due to nephropathies, blood loss or intestinal loss of protein, relative reductions in total protein only in Urea case of increased hydration. In cerebrospinal fluid, elevated levels are seen in case of inflammation as well as tumours associated Material S, EP, HP 0.5 ml with the brain. In urine, elevated levels are usually associated with Method Photometry glomerulonephritis. Species Dog, cat, rabbit, guinea pig, rat, ferret, reptiles, horse, ruminants, pig • Electrophoresis is used to separate the protein fractions. Duration 1 day 78 79 2019/20 Clinical Chemistry 2019/20 Clinical Chemistry LABORATORY FOR CLINICAL DIAGNOSTICS Note • Urea is the most important degradation product of the protein Note • Calcium measurement from EDTA plasma does not provide metabolism in mammals. Serum levels are not only influenced plausible values. by the renal function (excretion) but also by extrarenal factors • More than 99% of the body´s calcium is stored in the bones. Further (e.g. diet, increased protein degradation, liver function, metabolic functions are transmission of nerve impulses, muscle contractions, state). Thus, creatinine should always be checked in parallel. blood clotting etc. • In cattle, the urea level serves mainly as an indicator for energy • Non-parathyroid-related hypercalcaemia often occurs due to supply. tumours (in dogs mainly malignant lymphosarcoma). • Hypocalcaemia frequently is the cause of parturient paresis in cattle Uric Acid and of seizure disorders in small animals. • In case of concurrent hypoalbuminaemia, the calcium level should Material S, EP, HP 0.5 ml be corrected. Method Photometry Calculation: Corrected calcium level (mg/dl) = serum calcium level Species Dog, birds, reptiles (mg/dl) – (0.4 x serum protein (mg/dl)) + 3.3 Duration 1 day Note • Dog: Due to a metabolic disorder, increased levels of uric acid Calcium (Ca) ionised in the serum can occur especially in Dalmatians. Uric acid concrements in the urine and a characteristic brownish-yellowish Material S, HP 0.5 ml discolouration of the coat (Bronzing syndrome) are of clinical Method Potentiometry significance. Species Dog, cat, birds, reptiles • Birds: Concentrations above 500 μmol/l indicate nephropathies or Duration 1 day exsiccosis. Note • Ionised Ca represents the biologically active part of the total • Reptiles: The levels of uric acid vary greatly, depending on the calcium. last feed intake and the protein content of the ration. • The sample should be collected and stored without air (vacu tainer system). The instructions can be requested from us. β-Hydroxybutyrate (β-HBS) Material S, HP 0.5 ml Chloride (Cl) Method Photometry Material S, HP 0.5 ml Species Dog, cat, farm animals Method ISE Duration 1 day Species Dog, cat, rabbit, guinea pig, rat, mouse, ferret, birds, reptiles, horse, Note • Ketone bodies are formed in the organism during the degradation ruminants, pig of fatty acids. Duration 1 day • Ruminants: The determination of β-HBS provides an indication of Note • It is the most important extracellular anion and decisive for energy supply and can be used for diagnosing ketosis. maintaining the osmotic balance. • Increased levels are found in all diseases which also cause hypernatraemia. The most common causes are dehydration and hyperchloremic metabolic acidosis. 4.3 Minerals and Trace Minerals • Similarly, decreased levels occur in diseases which cause hyponatraemia, e.g. vomiting, abomasal reflux in case of Calcium (Ca) abomasal displacement, and metabolic alkalosis. Material S, HP 0.5 ml Method Photometry Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, horse, ruminants, pig Duration 1 day 80 81 2019/20 Clinical Chemistry 2019/20 Clinical Chemistry LABORATORY FOR CLINICAL DIAGNOSTICS Cobalt (Co) Magnesium (Mg) Material S, H 1 ml Material S, HP 0.5 ml Method ICPMS Method Photometry Species Horse, ruminants, new world camelids Species Dog, cat, rabbit, guinea pig, rat, ferret, horse, ruminants, pig Duration 1 week Duration 1 day Note • Central component of vitamin B12 (cyanocobalamin), which is Note Magnesium is essential for the energy metabolism of the cells and formed in ruminants by ruminal bacteria. for neuromuscular transmission. • Cobalt deficiency: reduced growth rate, milk yield and repro Hypermagnesaemia may occur in case of Addison’s disease; duction performance, coarse coat, cachexia, anaemia hypomagnesaemia is the most common cause of grass tetany in cattle and can also be found in case of renal dysfunction. Copper (Cu) Material S, HP 0.5 ml Manganese (Mn) Method Photometry Material S, HP 0.5 ml Species Dog, cat, horse, ruminants, pig Method AAS Duration 1 day Species Dog, horse, farm animals, others on request Note • Copper is a component of various enzymes. Decreased levels Duration 2 – 3 days can lead to depigmentation as well as growth and reproductive Note Determination is useful in case of suspected undersupply or disorders. intoxication. Undersupply may be caused by increased levels of iron • Dog: In the copper storage disease in Bedlington Terriers, copper in drinking water, as iron is antagonistic to manganese. Manganese serum levels usually are normal, elevated levels can only be deficiency in cattle mainly causes dysfunctions in skeletogeny and detected in liver tissue. Carriers and animals which have actually reproductive disorders. developed this disease can be identified by a genetic test. There are only few studies on the significance of manganese blood • Cattle: Decreased levels also occur in case of recumbency, levels in horses. anaemia and cardiac insufficiency. • Sheep: In newborn lambs, copper deficiency leads to CNS inorganic Phosphate (PO4) symptoms. Oversupply, e.g. caused by mineral supplement for cattle, leads to intoxication in sheep. Material S, HP 0.5 ml Method Photometry Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, horse, Iron (Fe) ruminants, pig Material S, HP 0.5 ml Duration 1 day Method Photometry Note • Increased values are found physiologically in young animals. Species Dog, cat, rabbit, guinea pig, rat, ferret, horse, cattle, goat, sheep, pig Due to high intracellular concentrations, haemolysis simulates Duration 1 day hyperphosphataemia. Preanalytically, haemolytic samples also Note • It is impossible to determine iron levels from EDTA plasma. show increased levels. • Iron is found in the body in the form of haemoglobin and • The most common causes of pathologically elevated serum levels myoglobin. Furthermore, it is a component of various enzymes. are kidney diseases and hyperthyreosis in cats. • Elevated iron levels in the serum occur due to the destruction • Endocrinopathies (diabetes mellitus, pseudohyperparathyroidism) of liver parenchyma (acute hepatitis, cirrhosis). In case of rarely may cause hypophosphataemia. occurring haemochromatosis and in connection with increased • Reptiles: The Ca/P ratio is approximately 2:1. serum levels, deposits are formed in the liver and muscles. • Cattle: Hypophosphataemia may lead to recumbency, amongst • Decreased levels are often associated with anaemia but also with others. Hyperphosphataemia occurs in case of acidosis. infections, malignant tumours and nephrosis. • Iron is a negative acute-phase marker. 82 83 2019/20 Clinical Chemistry 2019/20 Clinical Chemistry LABORATORY FOR CLINICAL DIAGNOSTICS Potassium (K) Sodium (Na) Material S, HP 0.5 ml Material S, HP 0.5 ml Method ISE Method ISE Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, horse, cattle, Species Dog, cat, rabbit, guinea pig, rat, ferret, birds, reptiles, horse, goat, sheep, pig ruminants, pig Duration 1 day Duration 1 day Note • Potassium is the most important intracellular cation. Note • Sodium is the most important cation of the extracellular space. In • Potassium levels are only significant if the sample (serum) has dogs and cats, sodium is excreted mainly by the kidneys. been freshly collected. • The main causes of hypernatraemia are loss of water without • Pseudohyperkalaemia may occur due to the release of potassium concurrent loss of electrolytes (diabetes insipidus, diabetes from platelets, leukocytes and erythrocytes. Preanalytically, mellitus), retention of sodium (mineralocorticoids) or a high sodium haemolysis also leads to falsely elevated levels. intake with food without the possibility of water uptake. • The main causes of hyperkalaemia are oliguria and Addison’s • The main causes of hyponatraemia are Addison´s disease, disease. diarrhoea, vomiting, or diuretics. • The main causes of hypokalaemia are vomiting or abomasal reflux, diarrhoea, renal dysfunction, Cushing’s disease/excess Sodium/Potassium Ratio (Na/K Ratio) glucocorticosteroids. • Cave: Even in case of absolute potassium deficiency, serum Material S, HP 0.5 ml levels may stay within reference range for some time! Methode Potentiometry • Cattle: Elevated levels of potassium may lead to reproductive Species Dog, cat disorders and recumbency if there is a relative sodium deficiency Duration 1 day at the same time. Note Slight hyponatraemia may be tolerated in small animals as long as the Na/K ratio is > 27:1. If the ratio is < 27:1, Addison’s disease is Selenium (Se) suspected (clarification by ACTH stimulation test). Material S, EP, HP 0.5 ml Method AAS Zinc (Zn) Species Dog, cat, horse, ruminants Material S, HP 0.5 ml Duration 1 – 2 days Methode Photometry Note • Selenium deficiency may cause nutritional muscular dystrophy Species Dog, cat, birds, horse, farm animals in foals. Special emphasis should therefore be placed on the Duration 1 day selenium supply of brood mares. Note • Marked zinc deficiency causes parakeratosis of the skin and • In cattle, it causes mastitis, puerperal disorder and abortion, in mucous membranes. In cats, changes in the hair coat are young cattle it causes growth disorder and paralytic myoglobin mainly being documented. Serum zinc levels are not necessarily uria. It leads to calf losses due to newborn myodystrophy, enzootic decreased in case of zinc responsive dermatosis. muscular dystrophy or white muscle disease. • Farm animals: Zinc deficiency leads to reduced feed intake and performance depression, changes in skin and claws as well as parakeratosis (especially in small ruminants). 84 85 2019/20 Urinalysis 2019/20 Urinalysis LABORATORY FOR CLINICAL DIAGNOSTICS 5 Urinalysis Fractional Electrolyte Excretion (FE) Material Urine 0.5 ml + S (non-haemolysed) 0.5 ml, samples collected at the For abbreviations and additional information concerning the test descriptions see page same time 11 and following. Method Photometry Species Horse Duration 1 day Kidney Profiles Horse Ø see Chapter 2.3.1 Note • The FE of Na, K, P, Cl are examined in horses. Kidney Pofiles Ruminants Ø see Chapter 2.4.1 • If the electrolyte excretion is put into relation with the creatinine excretion (here GFR = excretion), it indicates the FE of the Kidney Profiles Small Animals Ø see Chapter 2.1.1 electrolyte. • The FE is used to diagnose a dysfunction of the renal tubules. Kidney Profiles Pig Ø see Chapter 2.5.1 In horses with healthy kidneys, the net excretion of an electrolyte in the urine is regulated by the glomerular filtration rate and the Kidney-specific individual parameters that are determined from blood tubular reabsorption. If tubular reabsorption fails, the FE of one or (e.g. SDMA) Ø see Chapter 4.2 more electrolytes usually increases and its FE values will be above the normal range. COLA Test* Material Urine 3 ml γ-GT/Creatinine Ratio Method Column chromatography Material Urine 5 ml Species Dog Method Photometry Duration 7 – 10 days Species Horse Note • Quantitative determination of the amino acids cystine, ornithine, Duration 1 day lysine and arginine (“COLA”) in the urine. Note Shows the early stage of a tubular disease and is indicated in case • For the diagnosis of cystinuria in different breeds. of acute disease. • Elevated in case of nephropathy, glomerulonephritis and renal amyloidosis, among others Microalbumin • Additionally, a urine sediment and the determination of the pH value are recommended. Material Urine 0.5 m Method Photometry Species Dog, cat Fanconi Screening* Duration 1 day Material Urine 5 ml Note • The determination of microalbumin is considered a possibility to Method Column chromatography diagnose renal dysfunction in dogs and cats early. Species Dog • In contrast to the determination of the protein/creatinine-ratio Duration 7 – 10 days (U-P/C), this test is useful also in patients with no clinical signs. • Relatively unspecific test which may also yield positive results in Note • Quantitative determination of the amino acids threonine, case of inflammatory diseases (e.g. borreliosis, leishmaniosis). glutamine, proline, glycine, alanine and of the glucose • Sample must not contain blood admixture. concentration in the urine. • For the diagnosis of the Fanconi syndrome in dogs. • Additional examination of urinary sediment is recommended. 86 87 2019/20 Urinalysis 2019/20 Urinalysis LABORATORY FOR CLINICAL DIAGNOSTICS NABE (Net Acid-base Excretion) Urine Electrophoresis Material Urine 15 ml (fresh and cool) Material Urine 1 ml Method Titration Method Polyacrylamidgel Species Cattle Species Dog, cat, rabbit, guinea pig, ferret, horse, cattle, goat, sheep, pig Duration 2 – 3 days Duration 1 x per week Note Normal values of the fractionated net acid-base excretion (NABE) in- Note • Differentiation of nephropathies according to origin (e.g. tubular, dicate a physiological acid-base balance in cattle. NABE decreases glomerular). if feed intake is reduced. Together with the blood parameters ketone • Helpful in case of elevated protein/creatinine-ratio in the urine. bodies (ß-HBA) and free fatty acids (NEFA), NABE represents the • Not recommended if urine is contaminated with blood and in case minimal spectrum of metabolic control in cattle. of suspected prostatic cysts. • Detection of Bence-Jones protein in case of suspected myeloma. Protein/Creatinine-Ratio (U-P/C) Material Urine 1 ml Urine Sediment Method Photometry Material Urine 5 ml Species Dog, cat, horse Method Microscopic Duration 1 day Species Dog, cat, rabbit, guinea pig, horse, cattle, sheep, goat, pig, others Note • Parameter for early diagnosis of renal dysfunction in dogs, cats on request and horses. Duration 1 day • Not significant if the urine is contaminated with blood. Note • Diagnostic clarification of urinary tract diseases and superior • Increased values can also be caused by fever, bacterial and diseases (diseases of the liver, the kidneys or metabolic disorders) inflammatory processes without there being renal dysfunction. that can lead to changes in urination (polyuria, strangury and oliguria). Urinalysis incl. Sediment • Samples which arrive on Saturday will be analysed the same day. Delivery of the findings will follow on Monday. Material Urine 5 ml Method Mainly dry chemistry, photometry Species Dog, cat, rabbit, guinea pig, horse, cattle Duration 1 day Note Semiquantitative collection of clinical parameters and measurement of specific gravity. See Urine Sediment. Urinary Calculi Material Urolith (≥ 5 g) Method Infrared spectroscopy (FTIR) Species Dog, cat, reptiles, horse, cattle, sheep, goat, others on request Duration 1 day Note Chemical analysis of components in order to recommend specific diets as therapy and prophylaxis. 88 89 2019/20 Allergy 2019/20 Allergy LABORATORY FOR CLINICAL DIAGNOSTICS 6 Allergy Perennial Panel (horse) Material S 0.5 ml For abbreviations and additional information concerning the test descriptions see page Method ELISA 11 and following. Species Horse Duration 2 days Note Differentiation or detection of individual mould and mite allergens. 6.1 Allergy Testing Moulds: Alternaria alternata, Aspergillus fumigatus, Aspergillus niger, Cladosporium sp., Epicoccum sp., Helminthosporium sativum, Allergy Profiles (Pruritus Profiles Dog/Cat) Ø see Chapter 2.1.1 Penicillium sp., Fusarium sp., Ustilago sp., Rhizopus sp. Mites: Dermatophagoides farinae, Dermatophagoides Allergy Profiles (Pruritus Profiles Horse) Ø see Chapter 2.3.1 pteronyssinus, Acarus siro, Tyrophagus putrescentiae, Glycophagus domesticus, Lepidoglyphus destructor Allergy Screening Test Seasonal Panel (dog, cat) Material Dog, cat: S 2.5 ml Horse: S 1.5 ml Material S 1 ml Method Dog, cat: ELISA, Fcε-receptor technology Method ELISA; Fcε-receptor technology Horse: ELISA Species Dog, cat Species Dog, cat, horse Duration 2 days Duration 2 days Note Differentiation or itemisation of seasonal allergens. Note • Cost-effective screening test to determine for which allergen Pollen: 6-grass mix (orchard grass, perennial ryegrass, Timothy group the main test should be performed or whether it is already grass, meadow fescue, Kentucky bluegrass, meadow soft grass); possible to test again after cortisone administration. rye, mugwort, ragweed, English plantain, nettle, common sorrel, • Dog, cat: groups pollen, mites, moulds, flea saliva birch, hazel, willow • Horse: groups pollen, mites, moulds, insects Ideal testing time at the time of exposure (3 – 4 weeks after the onset • Ideal testing time at the time of exposure (3 – 4 weeks after the of the symptoms). onset of the symptoms). Includes CHO test and, if necessary, blocking of CCD. All samples sent in are stored by us for 14 days. Hence, in this time frame, if there has been a positive test result, it is possible to request Seasonal Panel (horse) further tests from a sample sent in for a screening test. Material S 0.5 ml Method ELISA Main tests allergy Species Horse Duration 2 days Perennial Panel (dog, cat) Note Pollen: 6-grass mix (orchard grass, perennial ryegrass, Timothy Material S 0.5 ml grass, meadow fescue, Kentucky bluegrass, meadow soft grass); Method ELISA; Fcε-receptor technology rye, mugwort, lamb´s quarters/goosefoot, English plantain, nettle, Species Dog, cat sorrel, dandelion, rape, ragweed, hazel, alder, poplar, birch, beech, Duration 2 days willow Note Differentiation or detection of individual mould and mite allergens. Moulds: Alternaria alternata, Aspergillus fumigatus, Cladosporium sp., Penicillium sp. Mites: Dermatophagoides farinae, Dermatophagoides pteronyssinus, Acarus siro, Tyrophagus putrescentiae 90 91 2019/20 Allergy 2019/20 Allergy LABORATORY FOR CLINICAL DIAGNOSTICS Further main tests Food Allergens Exotic (dog, cat) Feathers/Hairs/Epithelia Material S 0.5 ml Material S 0.5 ml Method ELISA (microarray) Method ELISA; Fcε-receptor technology Species Dog, cat Species Dog, cat, horse Duration 7 days Duration 7 days Note • Determination of antibody titres (IgG and IgE) against 15 „exotic“ Note Epithelia of cat, dog, rabbit, guinea pig, parrot feathers, feather mix alimentary antigens: trout, goat, camel, buffalo, quail, hermetia, sweet potato, sunchoke, buckwheat, bean, carrot, pumpkin, zucchini, pea, yeast Flea Saliva (IgE) • Basis for the specific selection of fitting dietary components for an Material S 0.5 ml elimination diet. Method ELISA; Fcε-receptor technology • All of the above-mentioned allergens can also be ordered Species Dog, cat individually. Duration 2 days Note A combination of flea saliva and recombinant flea saliva allergen is Food Allergens Extended used as an allergen. Material S 0.5 ml Method ELISA (microarray) Food Allergens Basic (dog/cat) Species Dog, cat Material S 0.5 ml Duration 7 days Method ELISA (microarray) Note • Determination of antibody titres against 8 single allergens which Species Dog, cat are fed rather infrequently. Duration 2 days Dog: horse, ostrich, boar, reindeer, amaranth, millet, kangaroo, Note • Dog/c at: determination of antibody titres (IgG and IgE) against parsnip 19 (dog)/16 (cat) most relevant alimentary antigens. Cat: horse, ostrich, boar, reindeer, amaranth, millet, deer, rabbit Dog: beef, pork, lamb, chicken, turkey, duck, soy, wheat, corn, • Basis for the specific selection of fitting dietary components for an rice, egg, milk, barley, potato, oats, whiting, salmon, rabbit, deer elimination diet. Cat: beef, lamb, pork, chicken, turkey, duck, potato, soy, wheat, corn, rice, egg, milk, salmon, tuna, whiting Food Panel (horse) • Basis for the specific selection of fitting dietary components for an Material S 1 ml elimination diet. Method ELISA Species Horse Food Allergens Cani-DIAL/Feli-DIAL Duration 7 days Material S 1 ml Note • Determination of antibody titres against 8 single alimentary allergens: Method Immunoblot soy, molasses, oats, corn, barley, wheat, barn, lucerne. Species Dog, cat • Basis for the specific selection of fitting dietary components for an Duration 2 weeks elimination diet. Note Test to determine appropriate commercial diets for dogs/cats with (suspected) food allergies. Feed, against whose proteins no anti Hymenoptera Panel* bodies are detected, are suitable as (elimination) diet. Material S 0.5 ml Either 5 or 10 commercial diets can be tested. Please choose the Method ELISA; Fcε-receptor technology diets to be tested from the list of testable diets. This list, further Species Dog, cat information and a submission form can be found on our website Duration 10 days (laboklin.com/vetinfo/further-lab-infos/allergies). 92 93 2019/20 Allergy 2019/20 Allergy LABORATORY FOR CLINICAL DIAGNOSTICS Note Detection of individual allergens of bee, wasp, hornet, paperwasp. • Seasonal allergens (Timothy grass, perennial ryegrass, Bermuda This service also includes the CHO test and, if necessary, blocking grass, yellow dock, English plaintain, mugwort, lamb´s quarter/ of the CCD (cross-reactive carbohydrate determinants) reaction. goosefoot, pellitory, dandelion, nettle, ragweed, olive, cypress, pine tree, plane tree, common privet, birch) Insect Panel (dog, cat) • Ideal testing time at the time of exposure (3 – 4 weeks after the onset of the symptoms). Material S 0.5 ml • Includes CHO test and, if necessary, blocking of CCD (cross- Method ELISA; Fcε-receptor technology reactive carbohydrate determinant). Species Dog, cat Duration 7 days Other allergens Note Detection of individual allergens of deerfly (Chrysops), mosquito Various other individual allergens can be tested on request. Please contact our allergy team! (Culex sp.), horse fly (Tabanus sp.), stable fly (Stomoxys sp.) and cockroach (Blatella germanica/Periplaneta americana) 6.2 Allergen-specific Immunotherapy Insect Panel (horse) Material S 1 ml Allergen-specific Immunotherapy (ASIT) Method ELISA Material Not necessary Species Horse Species Dog, cat, horse Duration 3 days Duration Approx. 2 – 3 weeks Note Detection of individual allergens of black fly (Simulium sp.), Anmerkung • Hyposensitisation to seasonal mosquito (Culex sp.), horsefly (Tabanus sp.), housefly (Musca or perennial antigens domestica), biting midges (Culicoides sp.). according to test outcome (serum test or intradermal Malassezia testing). Material S 0.5 ml • Please note: Food and Hyme- Method ELISA; Fcε-receptor technology noptera allergens cannot be Species Dog, cat added to the therapy! Duration 7 days • Therapy has to be carried out for at least one year, if successful, Note • Detection of a sensitisation (IgE) to Malassezia in dogs. it is continued for a lifetime • Can be added to the ASIT. (patient-specific solutions). • The composition of an ASIT can Mediterranean Panel also be done on the basis of any Material S 2 ml other test result (e.g. intradermal Method ELISA; Fcε-receptor technology testing). Species Dog, cat • A maximum of 8 allergens or Duration 7 days allergen mixtures per set; if more than 8 allergens/mixtures are Note Detection of individual perennial and seasonal Mediterranean needed, the allergens will be spread allergens in dogs and cats: over a double set (2 sets), for which • Perennial allergens (Mites: Dermatophagoides farinae, the double price of a single set Dermatophagoides pteronyssinus, Acarus siro, Tyrophagus is charged. putrescentiae • Please enclose a veterinary prescription with your order! Moulds: Alternaria alternata, Aspergillus fumigatus, Penicillium • The starter set will last for 6 months. notatum) 94 95 2019/20 Immunological Tests / Markers for Inflammation 2019/20 Immunological Tests / Markers for Inflammation LABORATORY FOR CLINICAL DIAGNOSTICS • The direct Coombs test is preferable to the indirect Coombs test 7 Immunological Tests/Markers for because of its higher significance. • The direct Coombs test will be performed if an EB sample is sent Inflammation in, the indirect Coombs test is performed if serum is sent in. For abbreviations and additional information concerning the test descriptions see page 11 and following. Coombs Test (indirect) Material S 0.5 ml Acetylcholine Receptor Antibodies Method Agglutination Species Dog, cat, horse Material S 1 ml Duration 1 day Method IFAT Species Dog Note • This test is used for the detection of autoimmune haemolytic Duration 1 day anaemia. • Positive reactions also occur in almost all infections with blood Note • This test is used to detect myasthenia gravis in dogs. In this parasites. disease, antibodies against acetylcholine receptors are formed. • The direct Coombs test is preferable to the indirect Coombs test It is characterised by weakness of the striated muscles which is because of its higher significance. enhanced by stress. • The direct Coombs test will be performed if an EB sample is sent • The weakness of the muscles may be generalised or locally in, the indirect Coombs test is performed if serum is sent in. limited to few muscle groups, such as those of the oesophagus (megaoesophagus). C-reactive Protein (CRP) Antinuclear Antibodies (ANA Test) Material S 0.5 ml Method ELISA Material S, EP, HP 0.5 ml Species Dog Method IFAT Duration 1 day Species Dog, cat, horse Duration 1 day Note Inflammatory mediator (acute-phase protein) that is used to diagnose non-obvious inflammation and for therapy monitoring. Note This test is used for the serological detection of autoimmune diseases (e.g. lupus erythematodes). If the test yields a negative result, a biopsy should be taken and examined, as the serological Haptoglobin testing can be negative especially in case of local changes. Low Material S, HP 0.5 ml positive titres may also occur in many general diseases. Method Photometry Species All Coombs Test (direct) Duration 1 day Material EB 0.5 ml Note Haptoglobin is an acute-phase protein. Its levels increase due to Method Agglutination inflammation, e.g. pneumonia and enteropathies. Haptoglobin is Species Dog, cat, horse much more sensitive than fibrinogen. Duration 1 day Note • This test is used for the detection of autoimmune haemolytic anaemia. • Positive reactions also occur in almost all infections with blood parasites. 96 97 2019/20 Immunological Tests / Markers for Inflammation 2019/20 Immunological Tests / Markers for Inflammation LABORATORY FOR CLINICAL DIAGNOSTICS Immune Status: Complete Blood Count, T-cells, B-cells, CD4+, CD8+, • Foals: The lack of maternal antibodies is one of the most Ratio CD4/CD8 important predisposing factors for infectious diseases in foals. IgG determination in the blood of newborn foals allows for a Material EB 3 ml (< 48 h) timely diagnosis – before clinical symptoms occur – as well as the Method Flow cytometry initiation of therapeutic measures. Species Dog, cat, horse Duration 1 day Immunoglobulin M (IgM) Note • The cellular immune status includes a complete blood count as well as the determination of the relative and absolute numbers of Material S 0.5 ml peripheral lymphocytes, B-cells (CD 21+), T-cells (CD 3+), Method Photometry T-helper cells (CD4+) and cytotoxic T-cells (CD8+) Species Dog, cat, pig and others (only on request) • In dogs, determination of the immune status can be helpful Duration 2 days for monitoring pyoderma (German Shepherd), demodicosis, Note Most notably, the importance of IgM lies in the mediation of the systemic lupus erythematodes, leishmaniosis and T-cell primary immune response. IgM is also involved in the secondary deficiency. immune response but its significance is lower. The secondary • In cats, the cellular immune status is used to determine the immune response is mainly mediated by IgG. current phase of the disease in FIV-positive animals. The test is also used in FIV-positive cats with stomatitis. Leukaemia Classification (Leukaemia Differentiation) • In horses, it is used to clarify frequent and prolonged infections. Material EB 2 ml (max. 48 h old) + blood smear Method Flow cytometry Immunoglobulin A Species Dog Material S 0.5 ml Duration 1 day Method ELISA Note • It is recommended to send in more sample material if possible. Species Dog Up to 5 ml of sample volume are required if the total leukocyte Duration 3 days count is low. Note In animal serum, IgA is found in lower concentrations than the other • With > 30.000 lymphocytes or positive clonality in the PARR test, immunoglobulins. It is considered the most important immuno leukaemia classification (leukaemia differentiation) may allow to globulin in the outer conjunctival secretion and in the urine, though, differentiate between the acute and the chronic form as well as and exists in secretory form. between B-cell and T-cell leukaemia. This provides an indication of prognosis and treatment. Leukaemia classification (leukaemia Immunoglobulin G (IgG) differentiation) is also part of the Leukaemia Profile. Material S 0.5 ml Leukaemia Profile Ø see Chapter 2.1.1 Method Capillary electrophoresis Species Dog, cat, horse, foal, cattle, calf, lamb, pig (only on request) Duration 1 day Rheumatoid Factor Note • IgG is the strongest immunoglobulin fraction in blood serum. Material S 0.2 ml The greatest significance of IgG lies in the antibody-mediated Method Haemagglutination immune response. Due to its small size, IgG can diffuse from Species Dog, cat the capillaries and, thus, has an additional relevance in immune Duration 1 – 2 days reactions in tissue spaces and on the body surface. Note This test can be used to determine rheumatic locomotor disorders • Young animals: At birth, foals, calves and piglets have only in dogs and cats. It should be performed during an acute episode marginal IgG content in the blood. They mainly absorb IgG via because serology might yield negative results in symptom-free the colostrum. IgG content is therefore an indicator for a sufficient episodes. supply of colostrum. 98 99 2019/20 Immunological Tests / Markers for Inflammation 2019/20 Immunological Tests / Markers for Inflammation LABORATORY FOR CLINICAL DIAGNOSTICS Serum Amyloid A (SAA) Material S 0.5 ml Method ELISA Species Dog, cat, horse, cattle Duration 1 day (cat, horse) 2 – 3 days (dog, cattle) Note • Inflammatory mediator (acute-phase protein) that can be used in dogs, cats, horses and cattle to diagnose non-obvious inflammation and for therapy monitoring. • For horses, SAA can also be requested in combination with the large screening. Serum Protein Electrophoresis Material S, EP, HP 1 ml Method Capillary electrophoresis Species Dog, cat, rabbit, ferret, birds, reptiles, horse, cattle, others on request Duration 1 day Note • Acute inflammation leads to an increase in the alpha- and/or beta-globulin fraction. Polyclonal hypergammaglobulinaemia is caused by infectious, immune-mediated or neoplastic diseases. Especially in case of feline infectious peritonitis (FIP) the test is used to support the clinical suspicion. • Albumin, alpha- and beta-globulin fractions are lowered in case of severe liver diseases. Thyroglobulin Antibodies Ø see Chapter 8 100 101 2019/20 Immunological Tests / Markers for Inflammation 2019/20 Endocrinology / Tumour Markers LABORATORY FOR CLINICAL DIAGNOSTICS Thrombocyte Antibodies 8 Endocrinology/Tumour Markers Material EB 0.5 ml (max. 72 h old) Method Flow cytometry For abbreviations and additional information concerning the test descriptions see Species Dog, cat page 11 and following. Duration 1 day ACTH (Adrenocorticotropic Hormone) Note • If possible, it is recommended to send more EB as the required sample amount depends on the total platelet count. Material EP 0.5 ml (centrifuge, pipette off and cool promptly after collection) • There are two possible ways for the development of thrombocyte Method CLIA AB: Species Dog, cat, horse, others on request – Autoantibodies against platelets are formed. Only in this case a Duration 1 day positive test result can be expected. Note • ACTH remains quite stable in EP. – In the course of another underlying disease, immune complex • Indication: diagnostic aid in suspected Cushing´s disease in formation occurs causing secondary damage to the platelets. horses, therapy monitoring of dopamine receptor antagonists. • Evaluation: ≤ 15% negative, 16 – 29% questionable, ≥ 30% positive AFP Ø see Tumour Marker AFP • Detection of thrombocyte AB is also part of the thrombocyte profiles. Aldosterone* Thrombocyte Profiles Ø see Chapter 2.1.3 Material EP 0.5 ml (cool) Method RIA Species Dog, cat Duration ≤ 7 days Note Diagnosis of hyperaldosteronism due to unilateral tumours of the adrenal cortex; frequently shown symptoms are hypertension, acute vision loss, hypokalaemic polymyopathy. Androstenedione Material S 0.5 ml Method ELISA Species Dog, ferret Test frequency 1 – 2 x per week Note Used for detecting an endocrine active hyperplasia/neoplasia of the adrenal gland in dogs and ferrets. Anti-Müllerian Hormone (AMH) Material S, HP 0.5 ml (cool) Method ELISA Species Dog, cat, horse, cattle Duration 1 day Note AMH is a highly sensitive marker for the diagnosis of granulosa cell tumours, cryptorchidism, differentiation neutered/intact. 102 103 2019/20 Endocrinology / Tumour Markers 2019/20 Endocrinology / Tumour Markers LABORATORY FOR CLINICAL DIAGNOSTICS NT-pro-BNP (Brain Natriuretic Peptide) Erythropoietin Material EP 0.5 ml (cool) Material S 0.5 ml (cool) Method ELISA Method ELISA Species Dog, cat Species Dog, cat, others on request Test frequency 2 x per week Test frequency 1 x per week Note • The serum concentration of BNP depends on changes in blood Note Used for the diagnosis of renal-related non-regenerative anaemia pressure in the ventricle. Its determination mainly serves the early and to confirm or exclude polycythaemia. diagnosis of dilated cardiomyopathy. The myoendocrine cells of the heart start to secrete BNP as soon as the myocardium exhibits fT3 and fT4 Ø see after T3 resp. T4 vasodilative wall tension. BNP increases the renal excretion of sodium and water, lowers the intracardiac pressure and has IGF (Insulin-like Growth Factor 1) vasodilatory effects. • This test is suitable as a screening test for elder patients or Material S 0.5 ml (cool) predisposed breeds (e.g. Doberman). Method CLIA ELISA (horse, cattle) CEA Ø see Tumour Marker CEA Species Dog, cat Horse, cattle on request Duration 1 day Cortisol Note • Secretion is stimulated directly by the somatotrophic hormone Material S, HP 0.5 ml (= growth hormone), IGF-1 can therefore be interpreted as Method CLIA equivalent to STH (single value or function test). Species Dog, cat, rabbit, guinea pig, rat, mouse, ferret, horse, cattle, others • Indications are growth disturbances in young animals, changes on request in coat structure, acromegaly in adult animals, therapy-resistant Duration 1 day diabetes mellitus in cats. Note Single parameter with limited diagnostic value. Depending on the • Single value: only slightly decreased in case of growth clinical issue, the following tests can be performed: disturbances; if the result is questionable, a function test should • ACTH stimulation test be performed (xylazine stimulation test/STH stimulation test). • Low dose or high dose dexamethasone suppression test • In cattle, IGF-1 is suitable as a laboratory parameter for the early • Dog: Single value determination has extremely low diagnostic diagnosis of ovarian cysts and laminitis. value because of episodic secretion. • Cat: High concentrations may occur due to stress, therefore, Inhibin B* single value determination has extremely low diagnostic value. Material S 3 ml (centrifuged, cooled!) Method RIA CPSE (Canine Prostate Specific Arginine Esterase) Species Horse Material S, EP, HP 0.5 ml (cooled + centrifuged) Duration 4 weeks (will be passed on to a partner laboratory) Method ELISA Note Because of the long testing time we suggest testing for the Anti- Species Dog, male Müllerian Hormone. Test frequency 1 – 2 x per week Note • For the early detection of a benign prostatic hyperplasia in male dogs. • The enzyme CPSE is secreted by the prostate cells under the control of sex hormones. In hyperplastic prostate cells, CPSE levels increase significantly. 104 105 2019/20 Endocrinology / Tumour Markers 2019/20 Endocrinology / Tumour Markers LABORATORY FOR CLINICAL DIAGNOSTICS Insulin LH (Luteinising Hormone) Material S, HP 1 ml (cool) Material S (only) 0.5 ml (cool) Method CLIA Method ICA Species Dog, cat, ferret, horse, cattle Species • Cat, dog (female) Duration 1 day Duration 1 day Note • In case an insulinoma is suspected; concentration is only Note • To confirm or exclude an ovariectomy in dogs and cats. diagnostically conclusive if glucose is determined at the same time. • Interfering factors: haemolysis, lipaemia, non-cooled samples. • The serum must be immediately centrifuged, pipetted and cooled • If several samples are sent in, each sample will be charged before shipping. separately. • 12-hour fasting period prior to sampling (shorter in horses). • Dog/cat: Determine insulin/glucose ratio or AIGR (amended insulin/ Normetanephrine/Metanephrine-Creatinine-Ratio* glucose ratio). An insulin/glucose ratio < 52 or an AIGR < 30 are considered normal. Material Urine 10 ml (cooled, light-protected, acidified) • Equine metabolic syndrome (EMS): EMS leads to a disorder of Method HPLC carbohydrate and fat metabolism with insulin resistance. Increased Species Dog insulin secretion (partly) compensates for decreased insulin Duration 10 days efficiency. Insulin-resistant horses therefore have considerably increased fasting insulin levels. At the same time, fasting glucose Normetanephrine/Metanephrine* is physiological (compensated) or elevated (not compensated). Material EP 5 ml (frozen!) Further tests: glucose stress test, combined glucose insulin test and Method HPLC glucose stress test by feeding (see functional tests, Chapter 9). Species Dog • Simultaneous measurement of the glucose level makes it possible Duration 10 days to calculate the proxies - insulin/glucose ratio, - reciprocal inverse square of insulin (RISQI)-“insulin sensitivity” Oestradiol-17β and Material S 0.5 ml - modified insulin-to-glucose ratio (MIRG)-“β-cell function Method ELISA (pancreas)”. Species Dog, cat, ferret, horse, cattle, others on request Test frequency 2 x per week Leptin Note • The test is performed in case of disorders of the sexual cycle Material S, HP 0.5 ml (repeated determination), diagnosis of ORS (ovarian remnant Species Cattle, test is performed only on request syndrome – dog), neoplasia of the ovaries, ovary cysts, suspected Sertoli cell tumour. Note The hormone leptin is mainly produced by adipocytes. It affects, • Permanently elevated levels can lead to thrombocytopenia and among others, feed intake, glucose metabolism and fertility. Based anaemia due to bone marrow depression (dog). on the concentration of leptin, an assessment of the energy status (negative energy balance) is possible in early lactation. 106 107 2019/20 Endocrinology / Tumour Markers 2019/20 Endocrinology / Tumour Markers LABORATORY FOR CLINICAL DIAGNOSTICS Oestrone Sulphate Parathormone-rP (Parathormone-related Protein)* Material S 1 ml, urine 5 ml Material EP 1 ml (cool) Method ELISA/RIA* Method RIA Species Horse, donkey, others on request Species Dog, cat Duration Mare: 3 days (blood) Duration 10 days Diagnosis of cryptorchidism/female donkey: 7 – 10 days (urine) Note • PTHrP is a parathormone-like protein. Note • Mare: To determine an intact pregnancy • The hormone is formed physiologically during growth and during • In pregnant mares, oestrone sulphate is secreted by the pregnancy. fetoplacental unit in increasing concentrations from the 50th day • Pathologically, it is secreted by some tumours, e.g. by some kinds onwards. The oestrone sulphate level drops to basal level after of lymphoma, lymphosarcoma and anal sac adenocarcinoma. abortion or resorption within a few days. Reliable diagnosis is possible from day 110. PMSG/eCG (Pregnant Mare Serum Gonadotropin resp. Equine Chorionic • Stallion (from 3 years of age): Diagnosis of cryptorchidism (horse Gonadotropin) only; please indicate sex on submission form). Because of the higher sensitivity and as it is independent of age, we recommend Material S, HP 0.5 ml the determination of the Anti-Müllerian Hormone (AMH) instead. Method ELISA Species Horse, donkey Test frequency Horse 1 – 2 x per week; donkey: 7 – 10 days* PAG (Pregnancy-Associated Glycoprotein) Note • Proof of gravidity in horses and donkeys between the 45th day Material S, HP 1 ml and the 100th day. Method ELISA • PMSG can be detected for a while after resorption or abortion, Species Cattle, sheep, goat although there is no live foetus anymore. Duration 1 day Note Can be used from the 28th day after service to determine pregnancy Progesterone in cows. Material S, HP 0.5 ml Method CLIA Parathormone (iPTH) Species Dog, cat, horse, cattle, sheep, goat, alpaca, others on request Material S 1 ml (cooled + separated) Duration 1 day Method CLIA Note • Control of luteal function. Species Dog, cat, others on request • Can be used during early pregnancy to confirm conception in Test frequency 2 x per week cattle, horse, sheep and goat. However, as there are increases Note • iPTH (intact PTH) is unstable. in progesterone in the regular cycle, this difference is only • Determination is used to diagnose hyper-or hypoparathyroidism. diagnostically useful during the cycle-dependent decrease in • Concentration should only be assessed when ionised Ca (and progesterone. In horses and cattle, only samples which were possibly phosphate) levels are determined simultaneously. taken on the 18th and 19th day after mating are meaningful • Possible causes for high levels of PTH are low levels of calcium, regarding pregnancy. e.g. in case of renal dysfunction, and disorders of vitamin D • Female dog: Determination of the ovulation time, determination metabolism. of the optimum breeding time, diagnosis of corpus luteum insufficiency (repeated determination), measurement of corpus luteum function. 108 109 2019/20 Endocrinology / Tumour Markers 2019/20 Endocrinology / Tumour Markers LABORATORY FOR CLINICAL DIAGNOSTICS 17 OH-Progesterone Somatotropin Material S 0.5 ml Material S, HP 0.5 ml Method ELISA Species Cattle, test is performed only on request Species Dog, ferret, others on request Test frequency 2 x per week Thyroid Profiles Ø see Chapter 2.1.1 Note • Clarification of steroid-producing adrenocortical hyperplasia and neoplasia in ferrets and dogs. T3 (total Triiodothyronine) • In female animals in the luteal phase, considerably high Material S 0.5 ml concentrations may be measured. Method CLIA • In case of doubt, it is necessary to perform a function test. Species Dog, cat, horse, others on request Duration 1 day Relaxin Note • Additional parameter for the diagnosis of hyperthyroidism or Material S, EP, HP 0.5 ml (cool) hypothyroidism as the peripheral transformation of T4 to T3 only Method ELISA takes place when necessary and T3 is only secreted thyroidally to Species Dog, cat a small extent. Duration 1 day • If presence of T4 AB is suspected. Note • Relaxin levels increase significantly from day 25 after conception • Therapy monitoring: blood sampling 3 hours after oral intake of T3 and can be used to confirm pregnancy or to distinguish medication (dog). pseudopregnancy from pregnancy. • Date of mating and date of conception need not be identical; if fT3 (free Triiodothyronine T3) the test yields a negative result during early pregnancy, a control Material S 0.5 ml examination one week later should be considered. Method CLIA • Prolonged storage, uncooled transport, non-centrifuged sample Species Dog, cat, horse, others on request material or high storage temperatures lead to decrease in Duration 1 day concentration. Note T3 in the serum is 99.7% reversibly bound to transport proteins. fT3 levels correlate with T3 secretion and metabolic activity. If the thyroid Saliva Cortisol gland is underactive, fT3 concentration and total T3 concentration Material Saliva are generally parallel. Measuring free T3 is indicated when changes Method ELISA in the concentration of transport proteins for T3 result in changes in Species Dog, cat, guinea pig, horse, cattle, others on request total T3 concentration. Thyroglobulin concentrations are generally Duration 5 days quite constant, but may be altered e.g. during pregnancy and Note • Only on request. steroid treatments. In these cases, free T3 levels remain unchanged. • Test vessels will be provided. • Also possible: ACTH stimulation test T4 (total Thyroxine) Material S, HP 0.5 ml resp. 0.4 ml (small mammals) Serotonin Method CLIA Material S 0.5 ml (cooled or frozen) Species Dog, cat, rabbit, guinea pig, ferret, birds, reptiles, horse, cattle, Method ELISA others on request Species Dog Duration 1 day Duration 10 days Note Part of the diagnostic procedure in cases of abnormal behaviour. 110 111 2019/20 Endocrinology / Tumour Markers 2019/20 Endocrinology / Tumour Markers LABORATORY FOR CLINICAL DIAGNOSTICS Note • Dog: For the diagnosis of hypothyroidism. As a single value, it Note • Used for checking the endocrine testicular function, for the has only limited diagnostic significance. Determination should diagnosis of ovarian tumours in mares and to differentiate therefore always be performed in conjunction with fT4 and TSH. between cryptorchid and castrated male animals. Alternatively, a TRH stimulation test can be performed. • The HCG stimulation test and, in horses, AMH (anti-Müllerian • Cat: For the diagnosis of hyperthyroidism, which is the most hormone) as well, are gold standard for the diagnosis of common hormonal disease in elder cats and which entails cryptorchidism. extensive sequelae (tachycardia, chronic diarrhoea, cachexia). Usually sufficient as a single parameter, in questionable cases, Therapy Monitoring Vetoryl® (pre pill) TSH can be determined additionally. • Therapy monitoring dog/cat: Blood sampling 4 hours after Material S 0.5 ml administration of thyroxine (dog) and 3 weeks after therapy start Method CLIA (cat). Species Dog • Horse: If (very rarely occurring) hypothyroidism is suspected, Duration 1 day determination of T4 and T3 with a subsequent TRH stimulation test is recommended. Therapy Monitoring Vetoryl® (pre-, post pill) Material S 2 x 0.5 ml fT4 (free Thyroxine T4) Method CLIA Material S (possibly HP) 0.5 ml or 0.4 ml in small mammals Species Dog Method CLIA Duration 1 day Species Dog, cat, rabbit, guinea pig, horse, others on request Duration 1 day Thyroglobulin Antibodies Note • Single value determination of fT4: fT4 and fT3 are both strongly Material S, EP 0.5 ml affected by the current metabolic state. Method CLIA • Just like T4, fT4 is affected by underlying diseases. Species Dog • Fasting for 10 hours prior to sampling is recommended. Duration 1 day • In cases of doubt: TRH stimulation test or, in dogs and cats, Note • For the diagnosis of autoimmune thyroiditis in dogs. measure TSH concentration. • Detection of thyroglobulin AB is also part of the thyroid profile (dog) and of: fT4 + TSH + Thyroglobulin Antibodies (dog), see fT4 Dialysis* Chapter 2.1.1. Material S 0.5 ml (frozen) Method Equilibrium dialysis Thyroid-stimulating hormone (TSH) Species Dog, cat Material S 0.5 ml Duration ≥14 days Method CLIA Species Dog, cat fT4 + TSH + Thyroglobulin Antibodies (dog) Ø see Chapter 2.1.1 Duration 1 day Note • Dog: For the diagnosis of hypothyroidism only useful if T4 or Testosterone fT4 are determined simultaneously, as TSH values are within the Material S, HP 0.5 ml normal range in > 25% of the dogs with hypothyroidism. Method CLIA, ELISA • Therapy monitoring during diagnostic treatment. The dosage of Species Dog, cat, rabbit, guinea pig, rat, mouse, ferret, horse, cattle, others thyroid hormones should be reduced if the concentration is on request < 0.03. Duration 1 day, resp. 2 x per week (horse) • Cat: For therapy monitoring. 112 113 2019/20 Endocrinology / Tumour Markers 2019/20 Function Tests / Calculations LABORATORY FOR CLINICAL DIAGNOSTICS Troponine I 9 Function Tests/Calculations Material S 0.5 ml (cool, centrifuged) Method CLIA For abbreviations and additional information concerning the test descriptions see Species Dog, cat, horse page 11 and following. Duration 1 day Preferably, the patients should have fasted for at least 12 hours prior to every function Note Acute damage of the cardiac muscle (highly specific myocardial test. Stress and excitement should be avoided. parameter); can be used for the diagnosis of hypertrophic cardio In horses, fasting is only necessary in exceptional cases! myopathy (HCM). ACTH Stimulation Test Tumour Marker AFP (Alpha Feto Protein) Diagnosis • Therapy monitoring in Cushing‘s disease Material S 1 ml • Initial diagnosis of Addison‘s disease Method CLIA • Iatrogenic Cushing‘s disease Species Dog, cat, others on request • Hyperadrenocorticism in ferrets Duration 1 day Species Dog, cat, horse, ferret, others on request Material S 2 x 0.5 ml Note • To some extent, slightly increased values also occur in case of Test procedure • First blood collection = baseline value benign liver diseases in dogs. • Dog: injection of 5 μg ACTH/kg as Synacthen® i.v./i.m. • In case of hepatopathies, values are not or only slightly elevated. • Cat: injection of 0.125 mg Synacthen®/animal i.v./i.m. • In case of liver tumours, values are slightly to significantly • Horse: injection of 100 I.U. ACTH i.v./i.m. (only hypoadreno elevated. corticism) • Values are increased physiologically during pregnancy. • Second blood collection 1 hour post ACTH injection = stimulation • Therapy monitoring: In case of preceding positive finding after value (dog, cat) surgical and/or chemotherapy, concentration should be within • Second blood collection 2 hours post ACTH administration = standard range. Recidivist checks (semi-annual). stimulation value horse Parameter Tumour Marker CEA (Carcinoembryonic Antigen) to be determined Cortisol Material S (possibly also EP, HP) 1 ml Interpretation • Addison‘s disease/iatrogenic Cushing‘s disease: cortisol Method CLIA concentration post stimulation < 10 ng/ml (20 ng/ml in 8% of the Species Dog, cat, others on request cases) Duration 1 day • Cushing‘s disease (dog): cortisol concentration post stimulation is more than 3 times the baseline value (baseline value in the middle Note • Values increase due to tumours of the gastrointestinal tract and of the reference range) or exceeds 150 ng/ml. the mammary glands, but also due to inflammatory processes. • Therapy monitoring: cortisol concentration post stimulation should • Therapy monitoring: In case of preceding positive finding after not exceed 18 – 73 ng/ml (dog)/15 – 50 ng/ml (cat) in therapy with surgical and/or chemotherapy, concentration should be within Vetoryl®. standard range. Recidivist checks (semi-annual). • Horse: In healthy animals, the cortisol level increases by approximately 80%; horses with hypoadrenocorticism show very low baseline values which do not or only slightly increase after stimulation. 114 115 2019/20 Function Tests / Calculations 2019/20 Function Tests / Calculations LABORATORY FOR CLINICAL DIAGNOSTICS ACTH Stimulation Test extended Bile Acid Stimulation Test Diagnosis • Endocrine-active adrenal neoplasia, adrenal hyperplasia Diagnosis Detection of a portosystemic shunt • Early Cushing‘s disease Species Dog, cat Species Dog Material S 2 x 0.5 ml Material S 2 x 0.5 ml Test procedure • First serum sample = value after fasting (10 h) Test procedure • First blood collection = baseline value • Feeding of 100 g meat plus 5 g fat/10 kg body weight • Dog: injection of 5 μg ACTH/kg as Synacthen® i.v./i.m. • Second serum sample 2 hours after feeding = post prandial value • Second blood collection 1 hour post ACTH injection = stimulation Parameter to value be determined Bile acids Parameter to Interpretation Stimulation values > 50 µmol/l are indicative of a portosystemic be determined Cortisol and 17-OH-progesterone shunt, stimulation values > 40 µmol/l are considered suspicious. Interpretation • In case of Cushing‘s disease (hyperadrenocorticism), the cortisol level post stimulation is over 150 ng/ml or 3 times the baseline Corrected Calcium Concentration level, as long as this is in the middle of the standard range. Chronic stress and other underlying diseases (e.g. diabetes Diagnosis • Hypercalcaemias that are not caused by hyperparathyroidism can mellitus) can also result in an abnormal ACTH response. be attributed to tumours. According to literature, a stimulation value of > 220 ng/ml is, at a • Hypocalcaemia often causes parturient paresis in cattle and very high percentage, associated with Cushing‘s disease. seizure disorders in small animals. • It should be noted that about 15% of dogs with pituitary • If hypoalbuminaemia exists, the calcium value should be hyperadrenocorticism and about 40% of dogs with adrenal corrected. hyperadrenocorticism show a normal increase, meaning it is not Species Dog, cat noticeably elevated. Material S • For the diagnosis of other steroidal adrenal pathologies, the Parameter Ca, total protein same samples can be used to additionally determine the 17-OH- Estimation Corrected Ca concentr. (mg/dl) = serum Ca (mg/dl) - progesterone level. This is also possible in case of questionable (0.4 x total protein (mg/dl)) + 3.3 results when determining the cortisol level after ACTH stimulation. • In dogs with physiological steroid hormone synthesis, the Cortisol Creatinine Ratio concentration of 17-OH-progesterone increases to up to 180 ng/dl Diagnosis Diagnosis of Cushing‘s disease including differentiation between in the ACTH stimulation test. adrenal and pituitary forms • Dogs with a possible imbalance in the synthesis, which is found, Species Dog for example, in case of enzyme defects or adrenal tumours, Material Morning urine 1 ml show an increased baseline concentration and significant Test procedure • Collection of morning urine day 1 = sample 1 hyperstimulation. • Collection of morning urine day 2 = sample 2 • Dogs with pituitary-dependent hyperadrenocorticism (PDH) also • Administration of dexamethasone on day 2: orally 3 x 0.1 mg/kg show hyperstimulation. If the cortisol level is also significantly bdw throughout the day increased after stimulation, it indicates classic HAC. • Collection of morning urine day 3 = sample 3 • Because of the test properties, patients that are already being Parameters to treated with Vetoryl® cannot be tested for 17-OH--progesterone. be determined Cortisol, creatinine Interpretation • Interpretation of the ratio of day 1 and day 2: < 15: Normadrenocorticism, Cushing’s disease is unlikely. > 15 and < 25: Questionable result > 25: Hyperadrenocorticism (Cushing’s disease) 116 117 2019/20 Function Tests / Calculations 2019/20 Function Tests / Calculations LABORATORY FOR CLINICAL DIAGNOSTICS • Interpretation of the ratio of day 3: Dexamethasone Screening Test (low dose) Dog (An increased ratio on day 1 and day 2 is a prerequisite) > 50% of the average value of the first two samples indicate an Diagnosis Screening in case of suspected Cushing`s disease adrenocortical tumour. The presence of non-suppressible pituitary Species Dog Cushing’s disease is possible. Material S 2 x 0.5 ml or 3 x 0.5 ml < 50% of the average value of the first two samples indicate pituitary Test procedure • First blood collection = baseline value Cushing’s disease. • Injection of 0.01 mg of dexamethasone/kg bdw i.m. or i.v.* • Blood collection 4 hours post injection = 1st suppression value • Blood collection 8 hours post injection = 2nd suppression value Parameter to be determined Cortisol Interpretation • Normal: baseline value in reference range or slightly elevated (due to stress), 4 hours post injection reduction by 50% or to < 10 ng/ml and 8 hours post injection to < 10 ng/ml. • Cushing‘s disease: baseline value in reference range or elevated and one or both suppression values > 10 ng/ml • Additional blood collection 4 hours p.i. gives additional information concerning the cause: • Pituitary: baseline value in reference range or elevated, 4-hour value reduced by 50% or to < 10 ng/ml and 8-hour value > 10 ng/ml. • Adrenal tumour: baseline value in reference range or elevated, no adequate reaction to the administration of dexamethasone after 4 and 8 hours. Dexamethasone Screening Test (low dose) Cat Diagnosis Screening in case of suspected Cushing‘s disease Species Cat Material S 2 x 0.5 ml or 3 x 0.5 ml Test procedure • First blood collection = baseline value • Injection of 0.01 mg of dexamethasone/kg bdw i.m. or i.v.* • Blood collection 4 hours post injection = 1st suppression value • Blood collection 8 hours post injection = 2nd suppression value Parameter to be determined Cortisol Interpretation • Normal: baseline value in reference range or slightly elevated (due to stress), 4 hours post injection reduction by 50% or to < 10 ng/ml and 8 hours post injection to < 10 ng/ml (> 1 µg/dl). • Cushing‘s disease: baseline value in reference range or elevated and one or both suppression values > 10 ng/ml • Additional blood collection 4 hours p.i. gives additional information concerning the cause: • Pituitary: baseline value in reference range or elevated, 4-hour value reduced by 50% or to < 10 ng/ml and 8-hour value > 10 ng/ml. 118 119 2019/20 Function Tests / Calculations 2019/20 Function Tests / Calculations LABORATORY FOR CLINICAL DIAGNOSTICS • Adrenal tumour: baseline value in reference range or elevated, no • An additional sample 4 hours post injection gives additional adequate reaction to the administration of dexamethasone after 4 information concerning a delayed decrease of cortisol. and 8 hours. • Blood sample 8 hours post injection = suppression value Parameter to be determined Cortisol Dexamethasone Screening Test (low dose) Horse Interpretation • Pituitary form: one or both values post injection < 10 ng/ml Diagnosis Cushing‘s disease (in rare cases both > 10 ng/ml) Species Horse • Adrenal: both values post injection > 10 ng/ml Material S 2 x 0.5 ml or 3 x 0.5 ml Test procedure • First blood collection = baseline value (blood sampling at around 4 – 6 pm) Glucose Tolerance Test • Injection of 40 µg/kg or 2 mg/50 kg of dexamethasone i.m. or i.v. Diagnosis Equine metabolic syndrome (EMS) • Blood sampling approx. 15 hours after administration of Species Horse dexamethasone (at around 8 – 10 am) = 1st suppression value – Material Each time 1 ml sodium fluoride blood may be omitted Test procedure • Fasting overnight • 2nd suppression value after approx. 20 hours (at around 12 noon) • In the morning: first blood collection = fasted glucose – obligatory • Directly followed by infusion of glucose: 0.5 g/kg bdw of a 50% • Because of the circadian rhythm, the indicated times of the day glucose solution slowly administered i.v. (within approx. 5 min) should be observed. • Blood samples 2 to 7 after 30, 60, 90, 120, 150 and 180 min Parameter to Parameter to be determined Cortisol be determined Glucose Interpretation • Cushing’s disease: one or both suppression values > 10 ng/ml Method Photometry • Cave: In late summer/autumn, healthy horses, too, possibly Interpretation The glucose level should be back at the fasted glucose level after suppress insufficiently. 3 hours. Note Cushing’s disease in horses is caused by “pituitary adenoma” For information regarding EMS see Insulin (Chapter 8). (hyperplasia of the pars intermedia). The hyperplastic cells have no cortisol receptors, that is why in horses suffering from Cushing’s Glucose Tolerance Test - Feeding disease the exogenous administration of dexamethasone does not suppress the endogenous secretion of corticoids as it does in Diagnosis Equine metabolic syndrome (EMS) healthy horses. Species Horse Material S, each time 1 ml (centrifuged, cooled) Test procedure Fasting overnight, in the morning Dexamethasone Suppression Test (high dose) (1) 1 g/kg bdw of glucose Diagnosis Differentiation between adrenal and pituitary Cushing‘s disease or Species Dog, cat (2) 0.5 g/kg bdw of glucose per os Material S 2 x 0.5 ml or 3 x 0.5 ml (can be done by the animal owner!) Test procedure • First blood collection = baseline value Blood collection after 2 hours • Injection of 0.1 mg (dog) or 1.0 mg (cat) of dexamethasone/kg Parameter to bdw i.m. or i.v. be determined Insulin Method CLIA Interpretation Healthy horses stay below these cut offs: (1) < 85 mU/l insulin (2) < 68 mU/l insulin EMS horses show higher values. 120 121 2019/20 Function Tests / Calculations 2019/20 Function Tests / Calculations LABORATORY FOR CLINICAL DIAGNOSTICS Combined Glucose-Insulin Test Parameter to be determined Oestradiol (bitch)/testosterone (male dog, male horse) Diagnosis Equine metabolic syndrome (EMS) Interpretation • Bitch: depending on cycle – significant increases are expected in Species Horse diestrus and late anoestrus. Material Each time 1 ml sodium fluoride blood • Intact male dog: post stimulation concentrations > 1 ng/ml of Test procedure Fasting overnight, in the morning: testosterone are expected. First blood collection = fasted glucose • Horse: depending on the clinical issue • Intravenous infusion of 150 mg/kg bdw of a 50% glucose solution • Directly followed by 0.1 units/kg bdw of insulin i.v. Blood samples after: 1, 5, 15, 25, 35, 45, 60, 75, 90, 105, 120, 135 HCG Stimulation Test Dog/Cat and 150 min Diagnosis • Detection of endocrine active tissue of the gonads (ovary, testis). Each time, the blood glucose level is measured. • Ovarian remnant syndrome (dog) Method Photometry • Cryptorchidism Interpretation In healthy horses, the glucose level should return to basal level Species Dog, cat within 45 min. Material S 2 x 0.5 ml Insulin-resistant horses: the time glucose needs to return to basal Test procedure • First blood collection = baseline value (male dog: testosterone, level is a measure for insulin resistance (quantification). bitch: oestradiol) Particularity: Determining the fasting insulin level from the baseline Injection of 500 IU of HCG (Ovogest®)/dog i.v. sample makes sense and can provide further insights. To do so, • Second sample after 1 hour = stimulation value (perhaps non-haemolysed serum would have to be sent in – parallel to the additional sample after 30 minutes) baseline NaF sample (see above). The same applies to the sample Parameter to after 45 min. At that time, the insulin level should be below be determined Testosterone (male) or oestradiol (female and male) 100 µU/ml. Interpretation • Male dog: post stimulation concentrations > 1 ng/ml of Cave: There is always a small risk of hypoglycaemia, thus, a testosterone indicate that there is testicular tissue. glucose infusion solution should always be kept at hand. • Bitch: depending on cycle – significant increases are expected in diestrus and late anoestrus. GnRH Stimulation Test Diagnosis • Detection of endocrine active tissue of the gonads (ovary, testis). HCG Stimulation Test Horse • Ovarian remnant syndrome (dog) Diagnosis • Detection of endocrine active tissue of the gonads (testis) • Cryptorchidism • Cryptorchidism Species Dog, horse Species Horse Material S Material S 2 x 0.5 ml Test procedure Bitch: Test procedure • First blood collection = baseline value (testosterone) • First blood collection = baseline value (oestradiol) • i.v. application of 5000 – 10000 IU of HCG (Ovogest®)/animal • Injection of 0.32 µg of GnRH buserelin (Receptal®)/animal i.v. • Second sample (1 hour post injection) = stimulation value • Second sample after 3 hours = stimulation value (testosterone) Male dog: Parameter to • First blood collection = baseline value (testosterone) be determined Testosterone • Injection of 0.32 µg of GnRH buserelin (Receptal®)/kg i.m. • Second sample after 1 hour = stimulation value Interpretation • Stallion: Testosterone concentrations between 0.05 and Male horse: 0.1 ng/ml after stimulation are considered as borderline and need • In the morning: first sample = baseline value (testosterone) further clarification – e.g. by determining the anti-Müllerian • Directly afterwards: intravenous injection of 0.04 mg of hormone. Higher values indicate the existence of testicular tissue. GnRH/horse • Second sample after 1 hour = stimulation value 122 123 2019/20 Function Tests / Calculations 2019/20 Function Tests / Calculations LABORATORY FOR CLINICAL DIAGNOSTICS Insulin Glucose Ratio • Second sample after 90 min = 1st stimulation value • Third sample 3 hours post injection = 2nd stimulation value Diagnosis Calculated parameter for the detection of an insulinoma Parameter to Species Dog, cat be determined fT4 Material S (centrifuged, cooled) Interpretation • Euthyroid: min. 1 x > 25 pmol/l Test • Questionable: min. 1 x 20-25 pmol/l, other samples < 25 pmol/l Evaluation • Ratio = (serum insulin (µU/ml) x 100)/(serum glucose (mg/dl)) • Hypothyroid: all samples < 20 pmol/l • Modified ratio (AIGR = amended insulin glucose ratio) = (serum insulin (µU/ml) x 100)/(serum glucose (mg/dl) – 30) Parameters to TRH Stimulation Test Dog II (2 x T4) be determined Insulin, glucose Diagnosis • Hypothyroidism Interpretation Ratio of < 52 and AIGR < 30 resp. are considered to be • The test serves as compromise between the single determination physiological. of hormones (T4, fT4 und TSH) and the TSH stimulation test. Species Dog STH (GH) Stimulation Test Material S 2 x 0.5 ml Test procedure • First blood collection = baseline value Diagnosis Determination of IGF 1 as indirect parameter for the secretion of • Injection of Thyroliberin® i.v. (100 µg up to 3 kg body weight, growth hormones, IGF secretion is stimulated directly by the growth 200 µg with a body weight > 3 kg) hormone (GH). • Second sample 4 hours post injection = 1st stimulation value • Changes are caused by STH deficiency and STH reactive • Determination of T4 in first and second sample dermatosis (without decrease of STH). Parameter to • Perform function test after exclusion of other endocrinological be determined T4 causes as reference range and pathological range overlap. Interpretation • euthyroid: elevation of T4 concentration by min 0.5 µg/dl to min Species Dog, cat 2.5 µg/dl Material S 0.5 ml (centrifuged, cooled) • questionable: elevation of T4 concentration by less than 0.5 µg/dl Test procedure • First blood collection = baseline value to > 2.5 µg/dl or by more than 0.5 µg/dl but to < 2.5 µg/dl • Injection of Xylazin (100 µg/kg) i.v. • Second blood collection after 30 minutes = stimulation value Parameter to TRH Stimulation Test Extended (2 x T4 + 2 x TSH) be determined IGF-1 Diagnosis Hypothyroidism Interpretation Significant increase is expected: Species Dog • > 2 times if baseline value is low Material S • > 1.5 times if baseline value is high Test procedure • First blood collection = baseline value Note The determination of IGF needs to be requested individually for • Injection of Thyroliberin® i.v. (100 µg up to 3 kg body weight, every blood sample and will be invoiced per sample. 200 µg with a body weight > 3 kg) • Second sample 20 min post injection TRH Stimulation Test Dog (3 x fT4) • Third sample 4 hours post injection = stimulation value Parameters to Diagnosis • Hypothyroidism be determined • T4 (samples 1 and 3) • The test is a compromise between the single determination of • TSH (samples 1 and 2) hormones (T4, fT4 and TSH) and the TSH stimulation test. Interpretation • T4 (see above) Species Dog • Test is of limited informative value if there is no TSH rise in sample Material S 3 x 0.5 ml 2 (= after 20 min) Test procedure • First blood collection = baseline value • Injection of Thyroliberin® i.v. (100 µg up to 3 kg body weight, 200 µg with a body weight > 3 kg) 124 125 2019/20 Function Tests / Calculations 2019/20 Vitamins LABORATORY FOR CLINICAL DIAGNOSTICS TRH Stimulation Test Horse I (2 x T4) 10 Vitamins Diagnosis Hypothyroidism Species Horse For abbreviations and additional information concerning the test descriptions see page Material S 11 and following. Test procedure • First blood collection = baseline value • Injection of Thyroliberin® 0.5 – 1 mg/horse i.v. (slowly) β-Carotene • Second sample 4 hours post injection = stimulation value Parameter to Material S, HP 0.5 ml be determined T4 Method Photometry Species All Interpretation Euthyroid: 2- to 3-fold rise after 4 hours Duration 1 – 2 days Note In cattle, low amounts of β-carotene are associated with reduced TRH Stimulation Test Horse II (2 x ACTH) fertility and decreased resistance with subsequent diarrhoea. Diagnosis Cushing‘s disease Test with high sensitivity and specificity for the diagnosis of Folic Acid Cushing’s disease If the results of ACTH determination or the suppression test do not Material S 0.5 ml correlate with clinical findings or are not conclusive Method CLIA Species Horse Species Dog, cat, others on request Material 2 x EP (centrifuged, cooled) Duration 1 day Test procedure • First blood collection = baseline value Note • Differentiation between malabsorption and bacterial overgrowth • Slow injection of 1 mg of TRH i.v. (small intestine): Malabsorption will cause lowered levels of both • Second sample 10 min post injection = stimulation value parameters, bacterial overgrowth will lead to elevated folic acid in Parameter to conjunction with lowered Vitamin B 12 levels. be determined ACTH • Haemolysis is seen as the cause for falsely elevated values of Interpretation Stimulation cut-off value: 100 pg/ml; patients with Cushing’s disease folic acid. clearly exceed this. Vitamin A Xylazin Stimulation Test Ø see STH Stimulation Test Material S, EP, HP 1 ml (cooled) Method HPLC Species Dog, cat, horse, cattle, others on request Duration 3 days Vitamin B1 Material EB, HB 1 ml (cooled) Method HPLC Species Dog, cat, ruminants Duration 5 days Note Due to the lack of stability of the samples, analysis must be requested within one day after the samples have arrived at the laboratory. 126 127 2019/20 Vitamins 2019/20 Vitamins LABORATORY FOR CLINICAL DIAGNOSTICS Vitamin B2 Vitamin E Material EB, HB 1 ml (cooled) Material S, EP, HP 1 ml (cool) Method HPLC Method HPLC Species Dog, cat Species Dog, cat, horse, cattle, sheep Duration 5 days Duration 3 days Note Due to the lack of stability of the samples, analysis must be Note Concentration in horses living in a stable 1 – 2 mg/l, when grazing requested within one day after the samples have arrived at the 2 – 3 mg/l; in cattle > 3 mg/l. laboratory. Vitamin profiles Ø see Chapter 2.1.1 (dog/cat) resp. Chapter 2.3.1 (horse) Vitamin B6 Material EB, HB 1 ml (cooled) Method HPLC Species Dog, cat Duration 5 days Note Due to the lack of stability of the samples, analysis must be requested within one day after the samples have arrived at the laboratory. Vitamin B12 Material S (or possibly HP) 0.5 ml Method CLIA Species Dog, cat, cattle, others on request Duration 1 day Note • Can be used to differentiate between malabsorption and bacterial overgrowth or abnormal bacterial colonisation of the small intestine. • To clarify the need for a parenteral substitution of B12 in exocrine pancreatic insufficiency. • Cattle: Synthesis of vitamin B12 in the rumen can only take place insufficiently if too little cobalt is ingested with the feed. Vitamin B12 deficiency leads to disorders of the energy metabolism with lack of appetite, apathy, growth and performance depression and anaemia as well as possible diarrhoea. Vitamin D3 (25 OH) Material S (or possibly HP) 0.5 ml (cool) Method CLIA HPLC (cattle, sheep)* Species List Dog, cat, ferret, birds, reptiles, cattle, sheep Duration 1 day Cattle, sheep: 5 days 128 129 2019/20 Drug Level 2019/20 Drug Level LABORATORY FOR CLINICAL DIAGNOSTICS 11 Drug Level Note Therapy monitoring: Determination is suitable for monitoring the phenobarbital and primidone therapy in dogs and cats. In dogs, For abbreviations and additional information concerning the test descriptions see primidone is immediately metabolised to phenobarbital. Determina- page 11 and following. tion should take place at the earliest one week after starting long- term therapy. Sampling can be done regardless of when the drug is administered. Cyclosporine Material EB 1 ml Especially for horses, there is a series of drug level tests available as part of Method CLIA the doping analysis. Species Dog, cat We will be pleased to answer any questions that you may have. Duration 1 day Note Therapy monitoring: The determination is suitable for the control of • Doping-relevant substances cyclosporine therapy in dogs and cats. • Antiphlogistics Screening • Glucocorticoid Screening • NSAID Screening Digoxin • Sedativa/Tranquilizer Screening Material S 1 ml • Stimulant Screening Method CLIA • Tricyclic antidepressants Species Dog, cat Duration 1 Tag Note Therapy monitoring dog and cat: Soonest 7 days after initial appli- cation and approx. 6 – 8 hours after the last medication. Potassium Bromide Material S, EP, HP 1 ml Method ICP-MS Species Dog, cat Duration 4 days Note Therapy monitoring for dogs and cats under potassium bromide treatment. For this purpose, the concentration of bromide is measured. Phenobarbital Material S 1 ml Method CLIA Species Dog, cat Duration 1 day 130 131 2019/20 Intoxication 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS 12 Intoxication 13 Infectious Diseases: Pathogenic Agents For abbreviations and additional information concerning the test descriptions see and Antibody Detection page 11 and following. For abbreviations and additional information concerning the test descriptions see page Heavy Metal Toxicity Screening 11 and following. Material S 2 ml + EB 2 ml + urine 5 ml The obligation to notify the authorities upon suspicion or upon diagnosis of a disease Parameter Arsenic, lead, cadmium, chrome, copper, manganese, mercury, applies to Germany. thallium, zinc Species Dog, cat, horse, cattle Duration 5 days 13.1 Viruses Lead 13.1.1 Adenoviruses Material EB, HB 1 ml Adenoviruses are non-enveloped double-stranded DNA viruses, which are charac- Method AAS terised by a high tenacity. They belong to the linear double-stranded DNA viruses. Species Dog, cat, small mammals, birds, reptiles Adenoviruses are strictly host-specific and only in exceptional cases infection of related Horse, cattle on request or unrelated animal species occurs. Adenoviruses mostly cause mild respiratory symp- Duration 1 day toms and are involved in many multifactorial disorders. Note Because it is stored in the bones, lead can only be detected in high- er concentrations in the blood in case of acute poisoning. Over 95% Dog of lead in the blood is bound in erythrocytes, so as test material, Hepatitis contagiosa canis (HCC) whole blood is absolutely necessary. An elevated iron level in the HCC is caused by the canine adenovirus 1 (CAV-1). The virus is shed in urine and serum is an additional indication of possible lead poisoning. faeces and transmission occurs directly or indirectly. After oronasal infection, the virus first multiplies in the tonsils and subsequently in the endothelium of the blood vessels, Poison Screening* in hepatocytes as well as in cornea and uvea. Deposition of immune complexes can Material Vomit/stomach contents, blood, urine result in glomerulonephritis and uveitis with a corneal oedema (“blue eye”). HCC can Method Gas chromatography/mass spectroscopy be acute or chronic. Especially in unvaccinated puppies, HCC can take a peracute or Species Dog, cat, birds, horse, farm animals, others on request acute course and can be fatal. Not only dogs, but also all other species of the family Duration 10 days Canidae are susceptible to an infection with CAV-1. Note Global screening test, detection of, for example, coumarin deriva- Infectious laryngotracheitis tives. Please give us a call before collecting samples for a poison The infectious laryngotracheitis is caused by the canine adenovirus 2 (CAV-2). The screening in birds. virus has a strong affinity to the epithelia of the respiratory tract and is a component of Medical history is needed; please add information about medication the “kennel cough complex”. prior to sample collection. Reptiles Thallium (Rodenticides) Adenoviruses, which have mostly been documented in lizards and snakes, play an Material Urine, stomach contents, hairs important role in reptiles. Literature particularly describes adenoviruses in bearded Method AAS dragons (Pogona). The clinical picture is often non-specific. In Pogona, mainly young Species Dog, cat, horse, cattle animals are affected. Clinical symptoms which are often seen are anorexia, apathy, Duration 1 week diarrhoea and opisthotonus. Boas, colubrids and vipers belong to the snake families Note Thallium is a cumulative cytotoxin that can cause systemic that are often affected. Gastrointestinal symptoms are very typical. The liver, too, is intoxication. Cases of acute intoxication can be detected using urine, very frequently affected. Transmission probably occurs through the faeces, but also a detection in case of chronic intoxication is done using hair. vertical transmission is being discussed. 132 133 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Adenovirus – Pathogen Detection 13.1.3 Arenaviruses Material Dog: CAV-1: EB 0.2 ml, tissue (e.g. liver), urine, (faeces) Inclusion body disease of boid snakes (IBD) is caused by arenaviruses and CAV-2: swab without medium (e.g. eye, nose pharynx), particularly affects boas and pythons. Clinical symptoms comprise tremor, bronchoalveolar lavage opisthotonus and loss of the reflex to turn. In young animals, it is often an acute Birds: swab without medium (cloaca or pharynx), tracheal lavage, infection with a mortality rate of nearly 100%. In adult animals, the course of the disease tissue (e.g. intestine or liver) is usually chronic and protracted. Early symptoms are a slight tremor of the head, Reptiles: swab without medium (cloaca), tissue (intestine, liver) apathy and less flickering of the tongue. Method Realtime PCR (dog), PCR (birds, reptiles) Progression of the disease is often faster in pythons than in boas. Many times, Species Dog, birds, reptiles regurgitation is the first clinical symptom in boas. In pythons, the typical course of Duration 1 – 3 days (dog) the disease is a stomatitis accompanied by progressive pneumonia, which, showing 2 – 4 days (birds, reptiles) signs of CNS disease, leads to death. Over the past years, an increase of the disease has been observed in boas, whereas it does not occur as often in pythons anymore. Adenovirus – Antibodies To date, little is known about the transmission in reptiles. Transmission through close Material S, EP, HP 0.5 ml contact as well as through mites is being discussed. In some cases at least, a vertical Method IFAT transmission from infected parents to young animals seems to occur. Species Dog, others on request Diagnosis is either made through the detection of the characteristic intracytoplasmic Duration 1 day inclusions in the tissues of affected animals or through the detection of reptarenaviruses using PCR. In both cases, detection is easier in boas; in pythons, inclusions as well as Note Differentiation between infection and vaccination can only be done the virus are often only found in the brain. Histologically, inclusions are commonly seen by testing serum pairs (taken minimum 2 – 3 weeks apart from each in the pancreas, liver, kidneys, oesophageal tonsils and the brain. The same organs are other). Because of extensive vaccination, the disease has become suitable for PCR virus testing. In live animals, inclusions and viral RNA can be detected extremely rare. through blood smears or whole blood as well as through biopsies of the liver, kidney or the oesophageal tonsils. Oesophageal swabs are also very useful for virus testing. 13.1.2 African Horse Sickness Virus (AHSV) Arenaviruses/IBD – Pathogen Detection African horse sickness (AHS) is an endemic viral disease in Equidae particularly in Material EB, tissue (e.g. liver, pancreas, kidney, CNS), swab without medium Central Africa; sporadic outbreaks have been observed in the Middle and the Near East (oesophagus) as well as in Southern Europe (export-relevant test). Generally, the disease is transmitted Method PCR by Culicoides spp. or by Culex, Anopheles, Aedes and ticks. All secretions, intestines Species Snake (boa, python) and the blood of infected animals are infectious. A distinction is made between a Duration 1 – 3 days subclinical, febrile form, a subacute cardiac form, an acute pulmonary form and a mixed Note The PCR detects different reptarenaviruses. These viruses of the form; CNS manifestation is rare. family Arenaviridae are associated with the inclusion body disease In Germany, the disease is reportable upon suspicion. (IBD) in boid snakes (boas, pythons). A negative test result does not completely rule out IBD, as there may be other virus variants, which have not yet been described, leading to this disease. 134 135 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS 13.1.4 Avipoxviruses 13.1.6 Borna-Disease-Virus Avipoxviruses are normally only known as pathogens which cause avian pox in birds. The Borna virus is a non-segmented, enveloped RNA virus which constitutes the only They occur in many different bird species. Susceptibility of domestic and wild birds to member of the family Bornaviridae. This virus triggers the so-called Borna disease in avian pox infections is only partly understood. Avipoxviruses are primarily transmitted mammals and the proventricular dilatation disease in birds. through insects and aerosols. Breeders also become infected through contaminated animals or food and possibly through blood-sucking parasites as well. Introduction into Mammals the population mainly happens when buying additional animals or following exhibitions. Numerous species of mammals are susceptible to this virus. It is of clinical relevance In wild birds, infection also occurs directly when picking each other’s beaks. particularly in horses, in cats (where the disease is also called “staggering disease”) There are different characteristic forms. The cutaneous form is the most common one and in sheep. and is characterised by papular efflorescences of the non-feathered skin areas (eyes, The virus has a strong neurotropism and triggers non-purulent meningoencephalitis, beak, comb, lower legs). Mild forms often develop benign skin tumours (head, legs) as associated with anorexia, apathy, somnolence and multiple neuronal dysfunctions. a result of the long convalescence period (weeks/months). Animals suffering from Borna disease develop motor and behavioural disorders. In The mucous form is characterised by similar lesions on the mucosa of the beak cavity, horses and sheep, in addition to the symptoms listed above, a lowered head posture, tongue, pharynx or larynx (fowlpox/diphtheria). The septicaemic form typically displays separation from the herd, empty chewing and salivation have been described and, at general symptoms such as ruffled feathers, somnolence, cyanosis and anorexia without a later stage, recumbency and flailing movements. Cats frequently suffer from hind-leg exterior pox lesions. Avian pox infections are usually not fatal (exception: canarypox ataxia and lumbosacral pain. => often fatal). In Germany, there is an obligation to notify the authorities when There is often little or no immune response, which makes it difficult to diagnose by avipoxviruses are detected. testing for antibodies. The incubation period is unknown. The progression of a clinically manifested infection is lethal (duration of the disease usually 1 – 3 weeks). Avipoxvirus – Pathogen Detection Clinically inapparent infections are also possible. It has been a controversial issue Material Tissue (skin lesions, pigeon: small intestine; canary: oesophagus) whether this virus also infects humans and is there linked to neuropsychiatric disorders. Method PCR A seasonal increase of the disease from March to September has been described in Species Birds horses and sheep; in cats, increases can be found from December to May. Duration 1 – 3 days The modes of transmission have not yet entirely been clarified; infection probably occurs through the nerve endings of the nasal and pharyngeal mucosa. Infections Note The diagnosis can also be made by the histological detection of from horse to horse (sheep to sheep, cat to cat) are experimentally possible, but very inclusion bodies. unlikely. Shrews constitute the virus reservoir. Birds 13.1.5 Blue Tongue Virus Proventricular dilatation disease (PDD) is a globally distributed, serious disease particularly affecting psittacines (large parrots) like macaws, amazons or grey parrots. The bluetongue virus, an Orbivirus, is transmitted by biting midges and causes In 2008, identification of the previously unknown avian Borna virus (ABV) in birds bluetongue disease in cattle, sheep and goats. Bluetongue disease first occurred infected with PDD was achieved for the first time. An etiological relationship could then in Germany in 2006. So far, serotypes 6 and 8 have been recognised in Germany. be proven in infection experiments. Bluetongue disease is characterised by fever, circulatory disorders, head oedema, and PDD either affects the gastrointestinal tract, the central nervous system or both areas. ulcerations of the mucosa of the head as well as the teats and the feet. The disease can This means, on the one hand, there may be digestive disorders such as diarrhoea, also be accompanied by severe pneumonia. In sheep, the mortality rate can be 50%. vomiting or regurgitation as well as anorexia and the excretion of undigested seeds in Llamas and alpacas may become infected, too. In Germany, there is an obligation to faeces. On the other hand, PDD can manifest itself through neurological dysfunctions inform the authorities. like ataxia and coordination disorders, tremor or paresis. Both complexes of symptoms are associated with depression, general weakness and excessive loss of weight. Blue Tongue – Antibodies In addition to peracute and acute deaths, especially in older birds, chronic progressions Material S 1 ml of the disease can also be observed. Moreover, clinically inapparent birds can be Method ELISA infected with the virus. Breeding flocks and new additions should thus be tested for an Species Ruminants infection with ABV. Duration 7 days 136 137 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Avian Borna viruses are RNA viruses which show high genetic divergence. A negative Bovine Respiratory Syncytialvirus – Pathogen Detection result does therefore not entirely rule out PDD, as there may be other virus variants, which have not yet been described, leading to this disease. Material Swab without medium (nose, pharynx), lavage sample, tissue The safest way to detect an ABV infection requires a combination of antibody and (e.g. trachea or lung) pathogen detection. In some birds, only viral RNA can be detected, in others only anti- Method Realtime PCR ABV antibodies are detectable, while others again react positively on both tests. Species Cattle Duration 1 – 3 days Borna Virus – Pathogen Detection Note Particularly in the first phase of infection, BRSV can be detected by means of PCR. Material Mammals: CSF 0.2 ml, tissue (brain), EB 0.2 ml (viraemia), Antibody detection is part of the Bovine Respiratory Profile 1, see intraocular fluid (horse), retina (horse) Chapter 2.4.3. Birds: Swab without medium (crop and cloaca), tissue (brain, gastrointestinal tract) Method Realtime PCR Species Cat, birds (especially large parrots), horse, sheep 13.1.8 Bovine Virus Diarrhoea Virus (BVDV) Duration 1 – 3 days Bovine viral diarrhoea virus is the causative agent of bovine viral diarrhoea/mucosal disease (BVD/MD) in cattle, two diseases that are prevalent worldwide. Sheep, goats, Borna Virus – Antibodies* wild ruminants and pigs are also susceptible to the virus. BVD virus has 2 genotypes Material S 0.5 ml (BVDV1 and BVDV2) and the biotypes cytopathogenic (cp) and non-cytopathogenic Method IFAT (ncp). Birds: ELISA Infection in cattle results in different symptoms depending on the time of infection. Species Mammals, birds Transient infections (temporary infections of already born animals) are often Duration 1 week asymptomatic, but may lead to diarrhoea, fever, cough and erosions of the mucous membrane particularly in calves, and to reduced milk yield, fertility disorders (repeat breeding, abortions) and malformations (e.g. oculo-cerebellar syndrome) in cows. 13.1.7 Bovine Respiratory Syncytial Virus (BRSV) Transiently infected animals temporarily shed the virus to a certain extent (nasal discharge, saliva, faeces, semen). The bovine respiratory syncytial virus (BRSV) is an enveloped RNA virus of the family Persistently infected calves (PI animals) develop during the infection of the mother Paramyxoviridae and causes respiratory tract diseases in cattle and sheep. It is mainly between the 40th and the 120th day of gestation, because the immune system of calves that develop this disease. Infection primarily occurs in the winter months and the calf does not recognise the virus as “foreign”. PI calves are usually born without is characterised by sudden fever, slight hyperpnoea, apathy, rhinitis and cough. Mild abnormalities and shed large quantities of the virus in all secretions and excretions bronchiolitis, multifocal lesions and interstitial pneumonia with syncytia formation will throughout their lives. PI animals are typically seronegative, but can also form develop. The duration of the disease is 3 – 10 days. In severe cases, it can result in antibodies after being infected with a heterologous BVD strain. death, otherwise the fatality rate is low. Persistent infections have been described; they If a PI animal is additionally confronted with a cp virus strain through mutation of the may be the reason for maintaining the infection within a herd. prenatally acquired strain or a new, postnatal infection, it will develop a severe and In cattle, BRSV is involved in the enzootic bronchopneumonia; it predisposes calves always fatal MD. and lambs to the adherence of Mannheimia haemolytica. In Germany, there is an obligation to notify the authorities upon suspicion. There are different vaccines available. However, reinfections may occur after some months. 138 139 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Bovine Virus Diarrhoea Virus (BVDV) – Pathogen Detection Note Differentiation between infection and vaccination is usually only possible by analysing serum pairs (taken 3 – 4 weeks apart from Material EB 0.2 ml, milk, faeces, tissue (ear tissue tag samples, spleen, CNS, each other). Therefore, detection by PCR is preferable. abortion material) Method Realtime PCR Species Cattle 13.1.10 Caprine Arthritris Encephalitis Virus (CAEV) Duration 1 – 3 days The pathogen which causes CAE is a retrovirus and belongs to the genus Lentivirus, Bovine Viral Diarrhoea Virus (BVDV) – Antibodies just like the maedi visna virus. It is a viral disease in goats which causes encephalitis, Material S, milk 1 ml arthritis and/or mastitis, depending on the age of the animals affected. Development of Method ELISA clinical symptoms is slow. Neurological changes and interstitial pneumonia will result Species Cattle in ataxia, lameness, paralysis and respiratory distress. Only about one third of the Duration 3 – 5 days seropositive animals will contract the disease. The main transmission route is an infection of newborn animals via the colostrum. A 13.1.9 Caliciviruses horizontal and intrauterine transmission is possible, but of secondary importance. Caliciviruses see also Ø European Brown Hare Syndrome (EBHSV) CAEV (Caprine Arthritis Encephalitis Virus) – Antibodies Ø Rabbit Haemorrhagic Disease (RHD) Material S 0.5 ml Method ELISA There are numerous strains of the feline calicivirus (FCV) with only slight serological Species Goat, sheep differences but large genetic divergence, resulting in wide variations in virulence. Duration 2 – 3 days Symptoms of FCV may vary from inappetence and fever to joint pains and muscular pain. In rare cases, interstitial pneumonia occurs. The typical proliferate and exudative Note Particularly in affected herds, detection serves to eliminate positive ulcers in the oral cavity are often aggravated by secondary bacterial infections, i.a. with carrier animals. Positive animals are considered to be infected and pasteurella. potentially shedding (especially when lactating), negatives should be checked regularly (at least annually) since recent infection or low antibody titres can mimic a free result. Calicivirus (cat) – Pathogen Detection Material Swab without medium (conjuctiva, oral cavity, pharynx), EB 0.2 ml (only during the viraemia phase) 13.1.11 Chronic Bee Paralysis Virus (CBPV) Method PCR Species Cat Chronic bee paralysis virus is an RNA virus which, so far, cannot be assigned to any Duration 1 – 3 days family. This virus affects adult bees. Infected animals are unable to fly, they crawl on Note Due to genetic divergence, not all strains can be detected by means the ground, often have a bloated abdomen and diarrhoea; the affected bees die 5 – 10 of PCR. Detection in the blood is only possible during the viraemia days after the onset of the disease. Loss of hair and an asymptomatic progression are phase. possible. Transmission occurs through bee faeces. Whether the Varroa mite is involved in spreading or in worsening the course of the disease is debatable. There are often self-healing processes in the colonies. If the progress is particularly severe, an artificial Calicivirus (cat) – Antibodies swarm can be created with brood hatched in an incubator. Material S, EP, HP 0.5 ml Method IFAT Species Cat Duration 1 day 140 141 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Chronic Bee Paralysis Virus – Pathogen Detection disease is accompanied by immune suppression, and organ alterations occur, particularly in the central immune system and the spleen. In addition to the clinically Material Bees (heads) manifest form especially in young pigeons, there is also a high number of subclinically Method Realtime PCR or persistently infected animals. Species Bees Duration 1 – 3 days Pig Porcine Circovirus 2 (PCV-2) Porcine circovirus type 2 (PCV-2) is associated with the so-called Post Weaning 13.1.12 Circoviruses Multisystemic Wasting Syndrome (PMWS). PMWS is usually observed in weaners and less frequently in suckling piglets. Affected animals show a progredient loss of Dog weight as well as respiratory disorders with coughing, which are often complicated by Canine Circovirus secondary bacterial infections. PCV-2 can be detected in the tissue of infected piglets Canine circovirus was first detected in canine blood samples in the USA in by means of PCR. In conjunction with PMWS, co-infections of PCV-2 with porcine 2012/2013 and was described in a dog with necrotising vasculitis and granulomatous parvovirus or PRRSV are being discussed. lymphadenitis. In a subsequent study, it was mainly found in faecal samples from dogs with diarrhoea. In 2014, it was detected in Italy and in 2015 in Germany as well. Circovirus – Pathogen Detection Circoviruses can also be found in healthy dogs; further studies will be necessary to PBFD (birds) clarify questions on the pathogenesis and epidemiology. PCV-2 (pig) Canine circovirus should be considered in the differential diagnosis in case of diarrhoea/vomiting, fatigue, hepatic diseases, haemorrhage and vasculitis. Co- Material Dog: faeces, EB 0.2 ml (viraemia), tissue (above all liver, lym- infections with other, mainly enteropathogenic agents are frequently observed. Similarly, phoid tissue, intestine, kidney) an infection with canine circovirus can further complicate other infectious diseases. Psittacidae: 2 – 3 feather shafts (recently extracted), blood (1 – 2 drops on a filter paper), (faeces) Psittacidae Pigeon: 2 – 3 feather shafts (recently extracted), swab without Psittacine Beak and Feather Disease (PBFD) medium (cloaca), blood (1 – 2 drops on a filter paper, PBFD is characterised by an impaired growth of the beak, the feathers and the claws. viraemia!), faeces, tissue (bursa Fabricii, spleen, liver) The disease is globally distributed; more than 40 species of macaws, agapornis, grey Pig: EB 0.2 ml, swab without medium (nose or pharynx), parrots, amazons and parakeets are affected. lavage sample (BAL), tissue (e.g. lung, trachea, abortion material or foetal organs) Nestlings mostly die peracutely, while the course of the disease is acute in fledglings. Method Realtime PCR Animals show clinical signs of lethargy, loss of appetite as well as vomiting and/or Species Dog, psittacidae, pigeon, pig diarrhoea. Death is possible within 1 – 2 weeks. Changes in the developing feathers Duration 1 – 3 days are pathognomonic – but usually only visible in chronic forms. Symmetric feather loss Note PBFD PCR does not detect circovirus infections of other groups of occurs or the feathers get stuck in the shaft and will then break off. Lesions on the beak birds. and, rarely, on the claws will only occur later on. Transmission of the virus mainly takes place horizontally. The virus is spread with the faeces, the shedding of developing feathers and with the crop content of feeding 13.1.13 Coronaviruses parent birds. Thus, nestlings can be infected very early on. Vertical transmission is also possible, but of secondary importance. Here, hatching birds are infected through egg Coronaviruses are enveloped RNA viruses which have club-shaped appendages on shells that are contaminated with circoviruses. their surface that look like a crown under the electron microscope and have thus given the virus family its name. As they are genetically highly variable, transmission of corona- Pigeon viruses to and among different species is possible. They belong to a large group Circovirus infections mainly occur in pigeons aged 6 weeks to 12 months (Young of RNA viruses that can cause respiratory and/or enteral diseases in various animal Pigeon Disease Syndrome). The clinical picture is non-specific; symptoms include species and in humans. lethargy, anorexia, diarrhoea, wasting and PBFD-like changes in the feathers. The 142 143 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Dog Diarrhoea and mild colic symptoms may also occur. Neurological abnormalities (ataxia, An infection with canine coronaviruses (CCoV, CCV) is usually asymptomatic or may depression, recumbency) have rarely been described, they were, however, secondarily lead to mild, non-haemorrhagic diarrhoea. In puppies, however, severe courses of the caused by hyperammonaemia. The blood count shows leukopenia (neutropenia/ disease with haemorrhagic gastroenteritis are possible. Loss of intestinal villi, flattening lymphopenia) and hypoalbuminaemia. of the epithelial cells of the small intestine and detachment of the goblet cells occur. The Infections with ECoV seem to be self-limiting but can be secondarily complicated (e.g. most noticeable symptoms are vomiting and watery to bloody diarrhoea, accompanied dehydration or intestinal displacement). Transmission is mainly via the faecal-oral route. by severe dehydration. The virus is excreted via the faeces; the duration of excretion is usually less than two weeks. Cattle and Wild Ruminants Bovine coronaviruses (BCoV) cause enteral and respiratory diseases in cattle and wild CCV is also infectious to other canids as well as to cats and pigs, but the pathogenicity ruminants. These include calf diarrhoea, winter dysentery in adult cattle and respiratory of these species is not yet known. diseases in cattle of different ages. Cat Pig Feline coronaviruses (FCoV) can be divided into two pathotypes: the weak to non- In pigs, coronaviruses cause highly contagious, sometimes epidemic “transmissible virulent feline enteric coronavirus (FECV), which infects the intestinal epithelial cells and gastroenteritis” (TGE). TGE virus presents an economic problem in all countries with is considered a mere “diarrhoea pathogen”, and the fatal feline infectious peritonitis virus intensive pig production. Loss of income occurs in the affected farms as a result of piglet (FIPV), which mutates in cats and can replicate massively in macrophages. losses, growth retention and reduced weight gain. If FIP develops, two different clinical manifestations can be observed: the wet exudative form and the dry granulomatous form. In the wet form, severe polyserositis develops An infection with porcine coronavirus leads to a local infection of the intestinal tract, with the formation of a highly viscous, yellowish, fibrin-containing ascites fluid. In the mostly in the jejunum and ileum. As the disease progresses, the villous epithelium is dry form of FIP, pyogranulomatous swellings form on the serosa and in the organs. This rapidly lost. Clinically, this is manifested in a watery malodorous diarrhoea. TGE is a mainly affects the liver, lungs and kidneys. Inflammatory nodules form on the spleen and notifiable animal disease. the lymph nodes. Cats often develop anaemia with icterus, emaciation and high fever. There may also be CNS symptoms and, due to the deposition of precipitates, uveitis. Coronavirus – Pathogen Detection A positive titre indicates that the cat had been in contact with coronaviruses. This is the case with most adult animals. In a clinically healthy animal and especially if these Material Cat: FECV: Faeces/FIP: aspirate 0.2 ml, CSF 0.2 ml, intraocular animals come from breeding or animal shelters, high titres are usually not important. fluid, tissue (e.g. kidney) They do not suggest that this cat will contract FIP. However, if the titre exceeds 1:400 it is Cattle: faeces, swab without medium (nose), tissue (e.g. lung) assumed that these cats also excrete coronavirus with the faeces. In animals suffering Dog: faeces from FIP, there are often only low to negative antibody titres. In this case, the antibodies Horse: faeces have been bound in immune complexes; thus, antibodies are no longer detectable. For Pig: faeces, tissue (e.g. intestine) further evaluation, a serum protein electrophoresis and the determination of the albumin- Method Realtime PCR globulin quotient can be included. Diagnostic information is provided by an increase in Species Dog, cat, horse, cattle, pig the gamma globulin fraction and an albumin globulin quotient below 0.6. The albumin Duration 1 – 3 days globulin quotient is 92% specific. However, liver diseases such as cholangiohepatitis Note In small animals, a pooled faecal sample is recommended, thus, should also be considered for differential diagnosis. increasing sensitivity. Horse Coronavirus (FCoV, FIP) – Antibodies Equine coronavirus (ECoV), a beta coronavirus, was first detected in the USA in 1999 in the faeces of a foal suffering from diarrhoea. Recent studies in the USA, Japan and Material Ascites, S, EP, HP 0.5 ml Europe confirm it is associated with fever, colic and diarrhoea particularly in adult horses. Method ELISA Infections caused by ECoV mostly occur in the cold season (November to May); it could Species Dog, cat, pig not have been shown yet that the respiratory system is involved. Duration 1 day Clinical symptoms include anorexia, lethargy, fever and changes in faecal consistency. Note A positive titre merely confirms contact with coronaviruses. 144 145 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS 13.1.14 Cytomegalovirus viraemia occurs. Depending on the ability of the immune system to produce neutralising antibodies, distemper can take a mild or fatal course. Cytomegaloviruses belong to the herpesviruses and have been detected in different rodents. They are considered strictly host-specific. In guinea pigs, salivary and lacrimal Distemper Virus – Pathogen Detection glands become inflamed and respiratory symptoms may occur as well. In rare cases, Material Swab without medium (eye, nose, pharynx or tonsil), EB 0.2 ml there might also be signs of paralysis. (viraemia), CSF, urine, faeces Method Realtime PCR Cytomegalovirus – Antibodies* Species Dog, ferret, big cats, racoon, other species Material S 0.5 ml Duration 1 – 3 days Method IFAT Species Guinea pig, rat, mouse Distemper Virus – Antibodies Duration 1 week Material S, EP, HP 0.5 ml Method IFAT Species Dog, ferret, big cats 13.1.15 Deformed Wing Virus (DWV) Duration 1 day Deformed wing virus (DWV) is an RNA virus from the genus Iflavirus, which causes Note Differentiation between infection and vaccination is only possible by wing deformities in adult honey bees. All developmental stages can be affected. If means of testing serum pairs. Titres in liquor always are indicative already the larva is infected, it will develop crippled wings, a bloated abdomen and for infections (no contamination with blood!). discolouration during metamorphosis, and the animal dies shortly after emerging. The virus causes a latent infection that persists. Thus, bees without any symptoms can still be carriers. The Varroa destructor mite is among the vectors of this virus and its move 13.1.17 Equine Infectious Anaemia Virus (EIAV) to other colonies poses a major problem in the spreading of the disease. There is no causal therapy. Equine infectious anaemia (EIA) is a worldwide distributed disease in equidae, caused by a retrovirus, with acute lethal to chronic recurrent forms. Characteristic signs are recurrent fever, anaemia, thrombocytopenia, oedema and considerable weight loss. Deformed Wing Virus (DWV) – Pathogen Detection Transmission takes place via infected blood, blood-sucking insects, iatrogenic through Material Bees infected injection equipment, but also intrauterine. Method Realtime PCR Once infected horses remain infectious and seropositive throughout their lives. All Species Bees horses older than 6 months that are seropositive are thus considered carriers; younger Duration 1 – 3 days horses can be seropositive through maternal antibodies. Normally, the incubation period is 1 – 3 weeks, but may also last up to 3 months. In Germany, there is an obligation to inform the authorities, as EIA is an epizootic 13.1.16 Distemper Virus disease! Canine distemper virus (CDV) belongs to the genus Morbillivirus (measles-distemper- Equine Infectious Anaemia Virus – Antibodies rinderpest group). All animals of the families Canidae (such as dog, fox, wolf), Procyonidae (like raccoon and panda) and Mustelidae (such as ferret, badger, marten), Material Agar gel diffusion test (Coggins test): S 0.5 ml tiger and lion can get infected. Distemper is enzootic worldwide. Transmission takes cELISA: S 2 ml place orally or airborne via secretions and excretions of infected dogs or clinically healthy Method Agar gel diffusion test (Coggins test), cELISA carriers. Intrauterine infections are also possible. Distemper is a febrile general disease Species Horse, other equidae with an acute to subacute course. A respiratory, an intestinal, a central nervous and a Duration Agar gel diffusion test (Coggins test): 3 days cutaneous form of the disease can be distinguished. cELISA: 1 day Virus excretion begins after approx. 7 days (up to 60 to 90 days p.i.), during which a typical cyclic infection with leukocyte-associated (possibly also non-cell-bound) 146 147 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Note First antibodies can be detected 2 – 3 weeks post infection. If the 13.1.19 European Brown Hare Syndrome Virus (EBHSV) results of the serological examination are negative but animals are suspected of being infected, the test should be repeated within 3 – European brown hare syndrome (EBHS), also called viral hepatitis of hares, is a disease 4 weeks. Please observe the legal requirements. of the hare species Lepus europaeus and Lepus timidus. The disease was first described in Scandinavia in the 1980s and has since occurred in numerous European countries; several cases have been reported in Germany as well. 13.1.18 Equine Arteriitis Virus The causative agent of EBHS is a calicivirus (genus Lagovirus), which only causes the disease in hares. As far as is known, rabbits (and other animal species, too) are not Equine Viral Arteritis (EVA) is a worldwide distributed, contagious viral infection of affected. The virus is shed in all secretions and excretions and is very environmentally equidae caused by the equine arteritis virus (EAV). Confirmed outbreaks seem to have stable. Transmission presumably occurs directly, particularly faecal-orally or indirectly increased in recent years. The majority of naturally acquired infections is subclinical; through contaminated water and feed. The disease is peracute to acute and is however, seroconversion still occurs. When clinical symptoms appear, they vary in type characterised by a very high morbidity and mortality rate (up to 100%). If at all possible, and severity: fever, depression, anorexia and peripheral oedema, conjunctivitis (“pink symptoms are rarely observed in free-living wildlife. They include: weakness, apathy, eye”), urticaria and abortion. In young animals, pneumonia and pneumoenteritis may disorientation, loss of shyness and movement disorders (e.g. paralysis of the hind also be seen. The virus is mainly transmitted through semen. Persistently infected legs). There is no known therapy. carrier stallions carry the virus in their accessory sex glands and intermittently shed it in the genital secretions. Geldings, prepubescent stallions and mares cannot be carriers. European Brown Hare Syndrome Virus (EHBSV) – Pathogen Detection Especially in animals with systemic disease, excretion can also occur through other body secretions, such as aerolised secretions of the respiratory tract, urine, abortion Material Faeces, tissue (e.g. liver), (urine) material, etc. Method Realtime PCR In Germany, when EVA is detected in equidae (horses, donkeys, etc.), there is an Species Hare (not rabbit!) obligation to notify the authorities. Duration 1 – 3 days FIP Ø see Coronaviruses Equine Arteritis Virus (EVA) – Pathogen Detection Material Swab without medium (conjunctiva, pharynx), EB 0.2 ml (viraemia!), sperm, urine 5 ml 13.1.20 Feline Immunodeficiency Virus (FIV) Method Realtime PCR Species Horse, donkey, other equidae Feline immunodeficiency virus (FIV) belongs to the family Retroviridae. It is closely Duration 1 – 3 days related to the human immunodeficiency virus (HIV) but is not infectious for humans. Since FIV is mainly transmitted through bite injuries, the prevalence of infected animals Equine Viral Arteritis Virus (EVA) – Antibodies is highest in the group of uncastrated male cats over five years. FIV infection is spread worldwide. The prevalence in Germany is at approximately 3 – 5.5%. The virus persists Material S 0.5 ml for life. It has a clear tropism for T lymphocytes and macrophages. Similar to the clinical Method VNT symptoms of HIV-infected patients, the course of FIV infection is often divided into four Species Horse stages, with the final stage also being referred to as AIDS. However, transitions are Duration 5 days smoother and the phase with no clinical signs is often longer than in humans. Detection Note This detection is mainly required for export. should be performed in chronic recurrent and treatment-resistant infections, particularly It may be necessary to test paired samples after 2 – 3 weeks. in the oral cavity and the respiratory tract. 148 149 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Feline Immunodeficiency Virus (FIV provirus) – Pathogen Detection develops. Anaemia occurs in almost 50% of FeLV-infected cats, and in about 8% of them will lead to death. This is predominantly aplastic anaemia caused by disturbed Material EB 0.2 ml erythropoiesis. A reduced number of reticulocytes indicates aregenerative anaemia. Method Realtime PCR, qualitative or quantitative* FIV infections are also associated with lymphoid leukosis or leukopenia. Species Cat Duration Qualitative PCR: 1 – 3 days Quantitative PCR: 7 – 14 days Feline Leukaemia Virus (FeLV provirus) – Pathogen Detection Note Quantitative PCR: estimation of provirus load (therapy monitoring). Material EB 0.2 ml, bone marrow Method Realtime PCR Species Cat Feline Immunodeficiency Virus (FIV) – Antibodies Duration 1 – 3 days Material S, EP, HP 0.5 ml Note Detection of provirus can confirm a positive antigen result. Latent Method ELISA infections can also be detected if no antigen is present in the blood. Species Cat Duration 1 day Feline Leukaemia Virus (FeLV) – Antigen Note If the result is positive, the cat must be regarded as FIV-infected. (It is a lifelong persistent infection.) Material S, EP, HP 0.5 ml In questionable cases (exclusion of false positive results), the test Method ELISA should be repeated after 2 – 4 weeks. Species Cat Duration 1 day Note In order to distinguish transient from persistent infections, a positive 13.1.21 Feline Leukaemia Virus (FeLV) detection should always be controlled. This can be done after 4 – 6 weeks at the earliest, but better after 16 weeks. FeLV is a retrovirus. Retroviruses are enveloped single-stranded RNA viruses. The As this is an antigen detection, a “cross reaction” in vaccinated cats retrovirus family is characterised by reverse transcriptase and a sometimes oncogenic can be excluded. potential. The reverse transcriptase “transcribes” the RNA of the virus into cDNA, which is then integrated into the genome of the host cell. This way, provirus is created, which can remain a permanent part of the genome and is responsible for the mostly persistent 13.1.22 Hantavirus infections. FeLV is transmitted directly from cat to cat. The main source of transmission is saliva. In rats and mice, hantaviruses lead to a persistent infection and it is assumed they are Bite injuries represent a large transmission potential because here the infectious saliva permanently excreted in the urine, as well as through faeces and saliva, without the can get directly into the bloodstream. In most cases, this results in an oropharyngeal animals showing any symptoms. Hantaviruses are specific to individual rodent species infection of the cat. The virus penetrates the mucous membranes and within two days and only very rarely spread to other species. Rodents are infected in the burrow, in multiplies there as well as in the tonsils and retropharyngeal lymph nodes. Through territorial fights or while rearing the young. infected lymphocytes and monocytes, the virus enters the bloodstream and further into It is a zooanthroponosis which leads to severe medical conditions in humans. In the bone marrow after about twelve days. Europe, the clinical picture is mainly dominated by fever and nephropathies (dialysis If cats are infected with FeLV, only a transient infection with short viraemia develops in required) or haemorrhages. approximately 45% of cases. The immune system is able to eliminate the virus. The cat does not fall ill. In about 30% of the infections there is a sufficient immune response to Hantavirus – Antibodies* prevent virus replication, but virus elimination is not possible. A latent infection occurs. Other infectious diseases or stress can lead to viraemia reactivation. Material S 0.5 ml In all other cases, persistent infection occurs which usually results in a severe and Method IFAT short course of the disease. The primary consequences of the infection include organ Species Rat, mouse damage due to virus replication. Mostly, a FeLV-associated bone marrow depression Duration 1 week 150 151 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS 13.1.23 Herpesviruses CNS symptoms are also observed. The disease particularly breaks out in stressful situations, e.g. capture and quarantine of imported birds, change of owner, hospitalisation, Herpesviruses cause epidemic as well as latent or persistent diseases in almost beginning of breeding or the onset of sexual maturity. Therefore, a suitable preliminary all animal species. The name is derived from the Greek word “herpein” (to creep). examination of birds that are to be integrated into the flock is recommended in order to Common to all herpesviruses is lifelong latency in the host organism. avoid posing a threat to the other birds. Herpesviruses dog (CHV) An examination for herpesviruses may also be appropriate for other animals with systemic The so-called “puppy death” in dogs is caused by canine herpesvirus-1. Puppies under diseases, diseases of the respiratory system, the liver or with skin lesions or lesions of the 3 weeks of age die of haemorrhagic systemic disease. There is massive lytic virus mucous membrane at the cloaca or around the beak. replication at a subnormal body temperature of 36 – 37 °C and death occurs within 48 In amazon parrots and cockatoos, psittacid herpesviruses can also be detected in hours. The morbidity rate is 100%, the mortality rate is almost 95%! papillomas in the throat and the cloaca. Older puppies usually show mild respiratory symptoms, that is why an aetiological involvement in kennel cough complex is attributed to CHV. Herpesviruses reptiles Adult animals usually go through clinically inapparent infections. CHV-1 leads to a Herpesvirus infections are most common in a variety of chelonians, including tortoises, latent infection; after a primary cell-lytic infection, the viruses retreat into the trigeminal terrapins and sea turtles. In the veterinary practice, herpesviruses of tortoises of the and lumbosacral ganglion cells. In stressful situations (e.g. birth or incipient lactation), genus Testudo play an important role. As this is a highly contagious virus infection, viruses may be reactivated and shed in oral, nasal and ocular secretions. Female dogs animals should be routinely examined for infection before being introduced into a can transmit the virus in utero to the foetuses; abortions and stillbirths are rare. In adult population. immunocompromised animals, a peracute course of the disease with fatal outcome is Clinical symptoms include nasal and ocular discharge, regurgitation, anorexia and possible. A diagnosis of breeding animals is recommended. lethargy. Necrotic plaques in the oral cavity and on the tongue are also typical. So far, 4 different types of herpesvirus, testudinid herpesvirus 1 – 4 (TeHV 1 – 4), Herpesviruses cat (FHV) are known in tortoises. In Europe, especially TeHV 1 and 3 are found. TeHV 3 has a The main symptoms of feline herpesvirus (FHV) are respiratory symptoms such as broad host range among tortoises and infections are usually associated with very rhinitis and sinusitis with ocular and nasal discharge. Conjunctivitis develops and high morbidity and mortality rates. TeHV 1 can mostly be detected in Russian tortoises often corneal ulcers are formed. The cats suffer from dyspnoea and lack of appetite. (Testudo horsfieldii). These are often diseases of individual animals, since TeHV 1 has a These symptoms normally abate after a relatively short period of time. A latent infection considerably lower tendency than TeHV 3 to spread in the population. Individual cases develops which may be reactivated under stress at any time. This usually leads to of TeHV 2 (especially in desert tortoises) and TeHV 4 (in African tortoises) have been recurrent rhinitis, which, however, is milder than in the primary infection. Complications detected in Europe in recent years. are rare in FHV infections. Sometimes, the ocular changes are severe and the cat can go In turtles, herpesvirus infections are mainly associated with hepatic inflammation. In live blind. Moreover, very young kittens with very high fever and general weakness may die animals, dry throat swabs, and in dead animals, liver samples can be examined by PCR. (fading kitten syndrome). Herpesviruses cause fibropapillomatosis in sea turtles. Virus detection by PCR is possible from altered tissue. Herpesviruses birds In lizards, herpesviruses are mainly seen in connection with oral lesions. There are many different herpesviruses that are found in birds, including commercial poultry, ornamental, wild and zoo birds. New viruses are also regularly found in these Herpesviruses horse (EHV) animal groups. Several herpesviruses have been described in parrots, too. The best- EHV 1 and 4 known and perhaps clinically most relevant one is psittacid herpesvirus 1 (PsHV-1). In horses, donkeys, mules and zebras, infections with EHV 1 as well as with EHV 4 PsHV-1 is responsible for Pacheco’s disease in parrots and is therefore also called are caused by droplet infection or direct contact. The severity of the clinical symptoms Pacheco’s virus. The clinical course depends on the genotype or serotype and the depends on the age and immune status of the infected animal. Particularly infections affected psittacine species. For budgerigars and cockatiels, mild to subclinical courses with EHV 1 are able to spread beyond the respiratory mucosa and cause severe with virus shedding are reported. In large parrots, such as macaws, amazon parrots, manifestations of the disease: abortions, perinatal foal death, neurological diseases. cockatoos or grey parrots, an infection often leads to death. If symptoms occur, they In case of foals infected with EHV 4, morbidity rates of up to 100% are possible, are usually unspecific and consist of anorexia, apathy and poorly developed feathers. especially during the weaning period. More than 80% of the isolates come from animals Changes in faeces and an increase in uric acid excretion may occur, too. Occasionally, with rhinopneumonitis. EHV 3, which does not cross-react with EHV 1 or 4, causes genital infections in horses. 152 153 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Once horses are infected with herpesviruses, they remain carriers of the virus throughout for years and represent a potential hazard in the trade with live fish in pond management their lives, and the virus can be reactivated endogenously under unfavourable conditions and hobby animal keeping. Immunisation by means of live attenuated vaccine is currently (stress etc.). Lymph organs and the leukocyte fraction are the main latency organs. If the rejected from a scientific point of view. vaccinated horses are also taken into account, seroprevalence in the horse population is In Germany, it is an epizootic disease that is notifiable upon suspicion! high. In recent years, EHV-1-associated neurological diseases, for which a “neurotropic” strain Herpesvirus – Pathogen Detection of EHV 1 is held responsible, have been reported with increasing frequency and severity BHV 1 (cattle)* of the clinical disease. This much-feared clinical picture is referred to as EHM (equine herpesvirus myeloencephalopathy). Material Dog: abortion material, tissue of dead puppies (lung, liver, kidney), swab without medium (nose, pharynx, eye, genital EHV 2 and 5 tract), EB (viraemia) The involvement of EHV 2 and/or EHV 5 in keratoconjunctivitis has long been suspected Cat: swab without medium (eye, nose, phayrynx, genital tract), and these viruses are indeed regularly detected in conjunctival swabs. In recent years, it EB (viraemia!), abortion material, tissue (e.g, kidney, liver) has increasingly been shown that EHV 2 and 5 are precursors of other viral and bacterial Birds: 2 – 3 plucked pinfeathers, blood (EB or 1 – 2 drops on infections of the respiratory tract. Especially in young animals, EHV 2 and/or 5 were a filter paper), swab without medium (triple/3-fold swab: detected in treatment-resistant, partly catarrhal-purulent, partly necrotising or abscessing eye + pharynx + cloaca), faeces, tissue (e.g. liver, kidney, bronchopneumonia. EHV 5 was recently presented as aetiological agent of “equine spleen) multinodular pulmonary fibrosis” (EMPF). Tortoise: swab without medium (tongue), tissue (liver, intestine, (CNS)) EHV 3 Turtle: swab without medium (pharynx), tissue (liver) Coital exanthema caused by equine herpesvirus type 3 (EHV 3), which only sporadically Ocean turtle: alterated tissue occurs in Germany, is a mildly progressing breeding infection in horses. Clinically, Lizard: swab without medium (lesions, pharynx), tissue (liver) blisters, pustules and erosions appear on the mucous membrane of the vestibulum, Horse: EHV 1 and 4: swab without medium (nose or pharynx), penis or prepuce as well as on adjacent skin areas. Healing takes place spontaneously bronchoalveolar lavage, aborted material incl. placenta, after approximately 2 – 3 weeks, but can be complicated by secondary infections. EB 0.2 ml (viraemia) (upon request, the detection from Transmission mainly occurs through mating, but is also possible through close contact buffy coat is also possible, in this case 5 ml EB is as well as rectal and vaginal examinations. Infected animals remain carriers of the virus required), CSF for life. EHV 2 and 5: swab without medium (eye); foals with respiratoric symptoms: swab without medium (nose or Herpesviruses cattle (BHV) pharynx), bronchoalveolar lavage EHV 3: swab without medium (lesions on vestibule, penis, Bovine herpesvirus 1 (BHV 1) is the causative agent of infectious bovine rhinotracheitis prepuce or surrounding skin), tissue (lesions) (IBR), also known as infectious pustular vulvovaginitis (IPV) and infectious Cattle*: swab without medium (eye, nose or genital tract), tracheal balanoposthitis (IBP). lavage, abortion material, tissue (e.g. CNS or tonsil) In Germany, it is an epizootic disease that is notifiable upon suspicion! Koi: tissue (e.g. gills, CNS, liver, spleen, skin or intestine), swab without medium (gills or skin) Herpesviruses koi (KHV) Method Realtime PCR/PCR (birds, reptiles) Koi herpesvirus is a highly infectious virus that has caused epidemic disease in carps Species Dog, cat, birds, turtle, tortoise, lizard, horse, cattle, koi (koi and common carps) in recent years, depending on the water temperature. Morbidity Duration 1 – 3 days and mortality rates can be as high as 100% within 1 – 2 weeks after the pathogen has 2 – 4 days (birds, reptiles) been introduced. The incubation period ranges from a few weeks to several months. It Cattle: 7 – 14 days depends on various external and internal factors such as stress and condition of the fish. Fish of all age groups are affected at water temperatures between 18 – 29 °C. Clinically, Note Herpesviruses usually produce only short-term viraemia, thus the main symptoms are gill necrosis, increased mucus production, haemorrhages of the detection in EB is limited to early acute phase. skin, liver, spleen and kidneys. Surviving fish probably remain latent carriers of the virus 154 155 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Tortoise: In case of a positive result, differentiation of the virus Note Glycoprotein B (gB) has both vaccine strains and field strains in the strain may be of clinical relevance because of different virion. There is a deletion of the gE gene in the vaccine virus and tendencies to spread in the population and for the therefore no glycoprotein E (gE) in the virion, while the field virus prognosis of an infection. Differentiation is possible on has gE in the virion. Antibodies against gE can therefore only be request. detected in field virus infections, but not after vaccination alone. Horse: Blood test only secondary and only in febrile phase. From a positive buffy coat result, the conclusion can be drawn Infectious Anaemia Ø see Equine Infectious Anaemia Virus that the horse has come in contact with herpesviruses – Infectious Anaemia Ø see Equine Arteritis Virus whether this is acute or happened longer ago can often not be interpreted. In case of young horses, a blood test can be useful. 13.1.24 Influenza Virus Herpesvirus – Antibodies Influenza A viruses belong to the family Orthomyxoviridae and are mostly found in Pachecovirus (birds)* humans, pig, poultry and horses, but also in many others such as birds or dogs. BHV 1 (cattle) Material S, HP 0.5 ml Horse Milk 1 ml (cattle) Equine influenza is caused by the subtypes A equi 1 (H7N7) and A equi 2 (H3N8), alt- Birds: S 0.2 ml hough H7N7 has not been very present anymore over the past 30 years. In susceptible Tortoise: S, HP 0.4 ml Equidae, an infection causes fever and a rough, dry cough. In unvaccinated populati- Method Dog, cat, horse: IFAT ons, the virus spreads quickly. Secondary bacterial infections with mucopurulent nasal Birds, tortoise: VNT discharge are frequent and mask the clinical picture, especially in partially immune Cattle: ELISA populations. Species Dog, cat, birds, tortoise, horse, cattle Duration 1 day Pig Birds: 7 days, tortoises: 7 – 14 days Pigs may not only become infected with porcine, but also with human and avian influenza Cattle: 3 days viruses and thus contribute to the creation of reassortant influenza viruses. The influenza pandemics in humans in 1918/19 and in 2009 were caused by pig influenza viruses. In Note Differentiation between infection and vaccination is possible by pigs, primary infections are usually linked to livestock transport. The infection spreads means of testing serum pairs or, in cattle, by determining the gE explosively in the population. glycoprotein. Tortoise: The test detects antibodies against TeHV 1 and TeHV 3. Horse: Testing of paired serum at an interval of 10 – 14 days. A Influenza A Virus – Pathogen Detection fourfold titre increase would be proof of an acute EHV infection. Material Swab without medium (respiratory tract), bronchoalveolar lavage Method Realtime PCR BHV 1: Species Dog, horse, pig, others (not birds) gB – Antibodies* Duration 1 – 3 days gE – Antibodies* Material S, EP 1 ml Influenza A Virus (horse) – Antibodies* Method ELISA Material S 0.5 ml Species Cattle Method HAH Duration 5 days Species Horse Duration 5 days Note Differentiation between infection and vaccination is not possible. 156 157 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS 13.1.24 Iridovirus 13.1.27 Maedi Visna Virus Iridovirus Ø see also Ranaviruses Maedi and visna are two different diseases in sheep that are caused by the same virus of the retrovirus family and belong to the so-called “slow virus diseases”. Invertebrate iridoviruses (IIV) particularly occur in insects. In addition, they are regularly Maedi (meaning dyspnoea) disease is characterised by shortness of breath and cough, found in lizards, where they might cause skin lesions. The detection of iridovirus may be which are caused by chronic progressive interstitial pneumonia. of interest if the mortality rate in the population of feeder animals or lizards is increased. Visna (meaning decay) is a slightly contagious but progressive disease of the central nervous system. The animals show paralysis because of a demyelination of the CNS as Iridovirus – Pathogen Detection well as increasing decay. Notifiable animal disease! Material Tissue (e.g. skin or liver), swab without medium (skin), feeder ani- mals (whole insects) Maedi-Visna – Antibodies Method Realtime PCR Species Reptiles (lizard), feed insect Material S 0.5 ml Duration 1 – 3 days Method ELISA Species Cattle, sheep Note As these viruses are frequently found in feeder insects, virus detec- Duration 5 days tion in pharyngeal or cloacal samples or in the intestine needs to be interpreted very carefully. 13.1.28 Myxoma Virus 13.1.26 Lymphocytic Choriomeningitis Virus (LCMV) Myxoma virus is the causative agent of myxomatosis in rabbits. It is a large, enveloped DNA virus and belongs to the genus Leporipoxvirus (family: Poxviridae). Despite its The main reservoir of LCMV, which belongs to the arenaviruses, is the house mouse. envelope, poxviruses are relatively stable in the outside world. However, inactivation can Cells infected with LCMV express antigens and are recognised by cytotoxic T lympho- easily be achieved with ordinary disinfectants. cytes. This lymphocyte activity also makes the blood-brain barrier permeable resulting The virus is very host-specific: The European rabbit and domestic rabbit breeds de- in meninges and neurons being damaged. scending from it are most susceptible, but American rabbit species and European hare Infection of adult mice leads to choriomeningitis. In contrast, an intrauterine or neonatal species can be infected as well. The virus is mainly transmitted through insects (gnats, infection generally causes an asymptomatic chronic carrier state in mice, with such ani- fleas – mechanical transmission), thus, the disease primarily occurs between the end mals forming immune complexes in the course of their lifetime that lead to glomerulo- of July and October. Transmission by direct contact is usually only important in cases of nephritis. In guinea pigs and hamsters, LCMV infections often progress subclinically, high population density. however, conjunctivitis, blepharitis, respiratory symptoms, tremor, seizures and paraly- After a primary virus replication in the mucous membranes of the head, the regional sis have been described. LCMV is transmitted diaplacentally and with all secretions and lymph nodes become infected. Subsequently, there is a cell-associated viraemia (lym- excretions. phocytes) and the virus spreads to nearly all organs. In humans, LCMV rarely leads to choriomeningitis; the infection is usually asymptoma- After an incubation time of 4 – 10 days, an infection with myxoma virus causes an acute tic or shows mild, flu-like symptoms. An infection in the second part of pregnancy can systemic disease with severe conjunctivitis and hypodermal oedema (especially in the cause severe foetal damage. facial and the anogenital region). Nodular tumours in the skin and subcutaneous tissue may also occur. Respiratory problems and dysphagia result in inappetence and anorexia. Lymphocytic Choriomeningitis Virus (LMCV) – Antibodies* The mortality rate is between 25 and 90%. Chances of full recovery are generally very Material S, HP 0.5 ml low. Seriously affected animals should be euthanised. Method IFAT Due to the high mortality rate caused by the disease, the virus was introduced into Species Guinea pig, mouse, hamster rabbit populations in Europe, Chile and Australia around 1950 for population control. It Duration 1 week has since been endemic in these countries. However, co-evolution of the virus and the Note Zoonotic disease! 158 159 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS rabbits has led to weakened virus strains and virus-resistant rabbits. Thus, the severity 13.1.30 Orthopoxviruses of clinical symptoms strongly depends on the virulence of the present virus strain and the susceptibility of the host. The genus Orthopoxvirus belongs to the family Poxviridae. Due to their structure and Vaccines are available for prophylaxis. their viral enzymes, these viruses have a special position within the viruses. Poxviruses are able to mature into infectious viruses in the cytoplasm of the host cell without the Myxomavirus – Pathogen Detection cell nucleus being involved. Poxviruses have a relatively large genome with a double- stranded linear DNA. Material Swab without medium (conjunctiva, nose or pharynx), tissue Orthopoxviruses have a broad host spectrum and can therefore be called cow pox, cat (e.g. conjunctiva, lung or kidney) pox, elephant pox or rat pox. Cattle, carnivores, rodents and humans are particularly Method Realtime PCR susceptible. Species Rabbit In Germany, the authorities must be notified. Duration 1 – 3 days Cat Myxomavirus – Antibodies* An infection with cowpox virus can cause cat pox in both cats and humans. Cats are Material S 0.5 ml usually infected by their prey animals such as mice and rats. The virus penetrates the Method IFAT skin through bite or scratch injuries, which are usually located on the head, neck or fo- Species Rabbit relimbs. To some extent, necrotising, extremely itchy smallpox appears at these sites. In Duration 1 week most cases, self-healing occurs after a few weeks, but in immunocompromised people and cats (e.g. FIV infection), a systemic infection with severe to fatal pneumonia can Newcastle Disease Virus Ø see Paramyxoviruses develop. The vaccination against human pox administered until the 1970s does not provide any protection against an infection, but seroconversion with the vaccinia virus used 13.1.29 Nidoviruses for vaccination can probably lead to an attenuated clinical picture. These vaccinations were discontinued in the mid-1970s and a more frequent occurrence of this infection Viruses of the order Nidovirales are large, enveloped, single-stranded RNA viruses. becomes more likely. Among others, this order includes the families Arteriviridae and Coronaviridae. The A PCR analysis of skin crust material can provide a quick and reliable diagnosis. Self- nidoviruses detected in snakes are most closely related to the family Coronaviridae, protection during sampling and treatment of an infected cat should not be neglected. subfamily Torovirinae. They are found in pythons and boas, and have most commonly In addition, veterinary personnel and, if necessary, the owners should be educated. In been detected in ball pythons so far. They are associated with pneumonia and most cases, if a person becomes infected with smallpox, it can be diagnostically evalu- stomatitis and seem to be important pathogens in different python species. ated whether or not the pet is a carrier. Rat Nidoviruses – Pathogen Detection The occurrence of cowpox virus infections in pet rats and the resulting transmission to Material Swab without medium (pharynx or trachea), tracheal lavage, tissue humans has only recently been described. The rats show necrotising lesions on limbs (e.g. lung or trachea) and in the head and tail area. Method PCR Species Python, boa (Pox) Orthopoxvirus – Pathogen Detection Duration 1 – 3 days Material Scurf Method PCR Species Cat, rabbit, rat, mouse, cattle and others Duration 1 – 3 days Note Zoonotic potential, take special precautions when handling the sample! Pachecovirus Ø see Herpesviruses 160 161 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS 13.1.31 Papillomaviruses 13.1.32 Parainfluenza Viruses Dog/Cat Dog Canine papillomatosis is a rare viral disease in dogs characterised by numerous benign Canine parainfluenza-2 virus (CPiV-2) plays a crucial role in acute infections of the upper warts (papillomata) in the head area. The causative agent is canine papillomavirus. respiratory tract in dogs, which are referred to as kennel cough. In kennels or similar Although papillomaviruses occur in many animal species and in humans, they are strictly facilities, antibodies can be detected in up to 70% of all animals. host-specific, so they do not pose a risk to humans or other animals. Papillomata are Infections solely with CPiV-2 usually only lead to a mild or clinically inapparent course of mainly observed in the oral cavity and are less frequent on the conjunctiva, the cornea the disease. Only if secondary infections with other viruses (mainly canine adenovirus-2/ and the eyelids. Warts are benign and usually heal without treatment within one to five canine herpesvirus-1), bacteria (Bordetella bronchiseptica, streptococci, staphylococci months. If feed intake is severely affected by them, surgical excision might be indicated. etc.) or mycoplasma occur and if husbandry and/or hygiene conditions are poor, the A current study has attested a good effectiveness to the administration of azithromycin. known severe courses of the disease develop. Papillomavirus – Pathogen Detection Canine Parainfluenza Virus (CPiV) – Pathogen Detection Material Tissue (skin) Material Swab without medium (nose, pharynx), bronchoalveolar lavage Method PCR Method Realtime PCR Species Dog, cat Species Dog Duration 1 – 3 days Duration 1 – 3 days Horse Canine Parainfluenza – Antibodies The equine sarcoid is one of the most common skin tumours in horses (about 2 – 12% Material S, EP, HP 0.5 ml of all horses are affected). The causative agent is bovine papillomavirus - especially Method IFAT type 1, more rarely type 2. The tumour cells are modified fibroblasts; the skin and Species Dog subcutaneous tissue are affected. Equine sarcoids are considered semi-malignant Duration 1 day tumours, i.e. they do not metastasise, but have a strong tendency to recur if surgical removal is incomplete. It is presumed that transmission mainly occurs through direct Note As a rule, vaccine and infection titres can only be differentiated by contact as well as flies and horseflies, but also indirectly through wound sites, saddles, testing serum pairs. blankets and cleaning utensils. The entire skin surface as well as certain blood cells are infected; moreover, the infection remains throughout life. The initial diagnosis is made at Cattle the age of 3 – 12 years. Bovine parainfluenza-3 virus (PI-3, BPIV-3) plays an important role in acute respiratory tract diseases in cattle, especially in the development of the multifactorial disorder BPV 1/2 (Bovine Papillomavirus 1/2, equine sarcoid) – Pathogen Detection enzootic bronchopneumonia. Monoinfections cause only mild symptoms or are clinically inapparent. Only secondary infections with other viruses (e.g. bovine adenovirus), Material Scrufs, hairs (with root), tissue (tumour) bacteria (pasteurella, mycoplasma) and factors that reduce resistance (cold weather, Method Realtime PCR stress, poor stable hygiene) lead to the development of severe symptoms in the form Species Horse of endemic bronchopneumonia. Parainfluenza-3 virus is shed with the nasal secretion Duration 1 – 3 days and the transmission is airborne. The clinical picture is characterised by fever, breathing Note Positive PCR results confirm the suspected clinical diagnosis. Gold difficulties and salivation. About 5% of the animals die within 3 – 4 days. Various vaccines standard for the diagnosis of equine sarcoid is the examination by are available, but reinfections can occur after a few months. histopathology. 162 163 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Bovine Parainfluenza Virus (BPiV-3) – Pathogen Detection Avian Paramyxovirus 1 (aPMV-1) – Antibodies* Material Swab without medium (nose or pharynx), lavage sample, tissue (e.g. Material S, HP, EB 0.2 ml trachea or lung) Method HAH Method Realtime PCR Species Birds Species Cattle Duration 7 – 10 days Duration 1 – 3 days Note Antibodies against bovine parainfluenza-3 virus (BPiV-3) can be Reptiles detected using the bovine serological respiratory profile (Listing ID Paramyxovirus/Ferlavirus 1240). Ferlavirus infections most notably occur in snakes. These infections are rarely found in lizards and chelonians. Vipers, elapids, colubrids, boas and pythons are particularly affected. Symptoms of the disease include nasal discharge, breathing with open 13.1.33 Paramyxoviruses mouth and breath sounds. In addition to respiratory changes, CNS symptoms are often observed. They include poor muscle tone, compulsive movements, head tremor Paramyxoviruses see also Ø BRSV and opisthotonus. Transmission can occur horizontally from one animal to another, by Ø Parainfluenza Viruses aerosols or through faeces. Ø Distemper Virus In live animals, the virus can best be detected by obtaining a tracheal wash sample or Ø Sunshine Coast Virus through a combined pharyngeal and cloacal swab. The most suitable organ samples are lung, followed by brain, pancreas as well as liver, intestine and kidney. Paramyxoviruses are enveloped, single-stranded RNA viruses. They mainly cause respi- ratory disorders in humans and many animal species, but are also the causative agents Paramyxoviruses/Ferlaviruses – Pathogen Detection of severe systemic diseases. Material Swab without medium (pharynx, cloaca), tracheal lavage, tissue (e.g. brain, lung, liver, kidney, pancreas, intestine) Birds Method PCR Avian Paramyxovirus 1 (aPMV-1, Newcastle Disease Virus) Species Reptiles (especially snakes, but also lizards and chelonians) Newcastle disease virus is an avian paramyxovirus which can infect many different Duration 1 – 3 days avian species. In fowl, Newcastle disease is also called atypical fowl pest. There are different pathogenic strains which produce clinical symptoms of varying severity, from subclinical to peracute diseases. Most notably, affected animals can develop Paramyxovirus/Ferlavirus – Antibodies* respiratory and CNS symptoms; loss of performance, diarrhoea and sudden deaths are Material S, HP 0.2 ml also possible. Newcastle disease virus is zoonotic and can cause conjunctivitis, fever, Method HAH headache and aching limbs in humans. Species Reptiles aPMV-1 is considered the cause of the Newcastle disease once it exceeds a certain Duration 1 week pathogenicity index. Newcastle disease is a notifiable disease. Poultry must be vaccinated. 13.1.34 Parvoviruses Avian Paramyxovirus 1 (aPMV-1) – Pathogen Detection Parvovirus is a very small non-enveloped DNA virus with extreme environmental stability. Material Swab without medium (cloaca, trachea), tissue (trachea, lung, brain, It can persist in the environment for up to a year and is also very temperature-resistant. liver, spleen) Animals become infected oronasally with parvoviruses. First, virus replication occurs in Method PCR the mucous membranes, then followed by viraemia. The lymphatic system and organs Species Birds become infected. Duration 1 week 164 165 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Various clinical forms of parvovirus infection can develop. The peracute form results in Parvovirus – Pathogen Detection death within a few hours, usually without any serious symptoms. The acute form, how- ever, is characterised by severe symptoms. High fever, severe bloody diarrhoea and Material Dog, cat (qualitative): faeces, EB 0.2 ml, dog: also tissue vomitus occur. Due to the high affinity of the virus to tissues with high mitotic activity, (e.g. intestine, heart) severe leukopenia occurs simultaneously. If the number of leukocytes falls below 2000 Dog (quantitative): faeces cells/µl, prognosis must be made carefully. Subclinically infected animals represent the Ferret: EB (viraemia), swab without medium (rectum), tissue (e.g. pathogen reservoir as they shed the virus via the faeces. spleen, lymph node or bone marrow) Pig: swab without medium (genital tract), EB, tissue (e.g. abor- Dog tion material) Method Realtime PCR/PCR (ferret) In dogs, parvovirus infection usually progresses as a cyclic systemic disease with a Species Dog, cat, ferret, pig manifestation in the intestinal epithelium and the resulting clinical picture of anorexia, Duration 1 – 3 days/3 days (quantitative) fever, vomiting and persistent bloody diarrhoea. The disease is most severe in puppies. Note • PCR can be positive up to four weeks after vaccination with live Cat vaccine. Feline parvovirus infection – panleukopenia – is a highly contagious systemic disease • In dogs, differentiation between vaccine strain (CPV 2) and field of felids. The mortality rate among unvaccinated animals is over 80%. strains (CPV 2a, 2b, 2c) is possible on request. Please note that Clinically, the disease is characterised by fever, diarrhoea, vomiting and dehydration. when using parvovirus vaccines containing CPV 2b as vaccine The blood count shows extreme leukopenia. A special case is the intrauterine infection. antigen (e.g. Virbagen Puppy 2b) we cannot differentiate between The mother cat is infected without showing any symptoms, but it leads to the abortion vaccine strain and field strain. or death of the kittens. If kittens are born alive, there is often a cerebellar hypoplasia, • Direct detection of parvoviruses in the blood is possible approx. which leads to ataxia and tremor, usually without any impairment of consciousness. 1 – 5 days after infection. • Ferret: Rectal swabs lead to a much better sensitivity than faeces. Ferret Aleutian mink disease is caused by a parvovirus, genus Amdoparvovirus. This single- Parvovirus (cat, dog) – Antigen stranded DNA virus is non-enveloped and therefore, like canine and feline parvoviruses, Material Faeces extremely resistant. Minks, but also ferrets, skunks, otters, raccoons, foxes etc. can be Method EIA affected by this disease. Species Dog, cat The virus triggers an immune complex-mediated disease which is mainly characterised Duration 1 day by hypergammaglobulinaemia. The symptoms vary: Young animals tend to develop Note A faecal sample of 2 cm of diameter or larger is required. pneumonia, adult animals develop glomerulonephritis, arteritis, and/or meningoence- False positive reactions may occur up to 5 – 12 days post vaccination! phalitis. Hind leg paresis and fertility disorders have further been described. The outcome is often lethal. As there is currently no vaccine available, many ferrets are vaccinated with dog vacci- Parvovirus – Antibodies nes; it is unlikely that this will provide protection against an infection with the Aleutian Material Dog, cat: S, EP, HP 0.5 ml mink disease virus. Pig: S 1 ml Method Dog, cat: IFAT Pig Pig: ELISA Porcine parvovirus (PPV) can be detected in almost all pig populations worldwide. In Species Dog, cat, pig Germany, a prevalence of 60 – 80% can be assumed. Duration 1 day In an infection with PPV, fertility disorders and embryonic infections with subsequent Pig: 2 – 3 days fetal death (SMEDI: stillbirth, mummification, embryonic death, infertility) are the main Note Seroconversion occurs 4 – 7 days after infection; vaccine and clinical symptoms. The sows usually show no clinical signs. infection titres can only be differentiated by testing serum pairs. PBFD Ø see Circoviruses 166 167 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS 13.1.35 Picornaviruses Polyomavirus – Pathogen Detection Picornaviruses see also Ø Sacbrood Virus Material 2 – 3 freshly picked pinfeathers, blood (EB or 1 – 2 drops on a filter paper), faeces Picornaviruses are non-enveloped RNA viruses and in tortoises also known as virus Method Realtime PCR „X“. They are detected regularly in tortoises, and found associated with other infectious Species Birds pathogens, particularly with herpesvirus and mycoplasma. Duration 1 – 3 days Clinically, picornavirus is seen in young animals associated with a softening of the carapace. In adult tortoises, infections are seen as rhinitis, stomatitis, ascites and sudden death. 13.1.37 Porcine Respiratory and Reproductive Syndrome Virus (PRRSV) Picornavirus (Virus „X“) – Pathogen Detection Material Swab without medium (pharynx), tissue (e.g. intestine, tongue, Today, PRRS - also called swine infertility and respiratory syndrome (SIRS), porcine kidney, liver) epidemic abortion and respiratory syndrome (PEARS), mystery swine disease (MSD), or Method PCR blue ear disease - is among the world‘s most important diseases in pig production. In Species Tortoise Germany, the disease first occurred in the winter of 1990/91. Duration 1 – 3 days The virus can spread very quickly through droplet and airborne infection. The disease is characterised by late abortions around the 110th day of gestation. Dead or weak piglets can be born. In addition, there may be respiratory tract infections. Picornavirus (Virus „X“) – Antibodies Material S, HP 0.2 ml PRRSV – Pathogen Detection Method VNT Species Tortoise Material Swab without medium (nose or pharynx), EB, tissue (e.g. abortion Duration 1 – 2 weeks material, lung, trachea or lymph node), bronchoalveolar lavage, sperm Method Realtime PCR Species Pig 13.1.36 Polyomaviruses Duration 1 – 3 days Polyomaviruses are non-enveloped DNA viruses with a diameter of 45 nm and an Note Detection by PCR allows for a reliable diagnosis and the icosahedral capsid (similar to papillomaviruses). Polyomaviruses are latently present in differentiation between EU and US strain. mammalian cells and mostly cause persistent infections there. They usually form typical intranuclear inclusions in infected cells and after infection of heterologous cells lead to PRRSV – Antibodies* their transformation. They are therefore regarded as oncogenic. Polyomaviruses have a Material S 1 ml circular double-stranded DNA. Method ELISA Species Pig The highly contagious budgerigar fledgling disease virus (BFDV) is considered the Duration 3 days first avian polyomavirus (APV). BFDV causes an infection that may be fatal for psitta- cine nestlings; in adult birds, septicaemia and hepatitis are observed. In chronic cases, feather malformation and inability to fly occur; affected animals usually hop or run around. Particularly budgerigars are affected. The disease is also called French moult. 168 169 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS 13.1.38 Rabbit Haemorrhagic Disease Virus (RHDV) Rabbit Haemorrhagic Disease Virus 1+2 (RHDV 1+2) – Pathogen Detection Material Swab without medium (conjunctiva), urine, faeces, EB, bone Rabbit haemorrhagic disease (RHD), also known as rabbit calicivirus disease or viral marrow, tissue (e.g. liver) haemorrhagic disease, is a highly contagious disease of European rabbits (Oryctolagus Method Realtime PCR cuniculus). It occurs in both wild and domestic rabbits and causes peracute, acute or Species Rabbit subacute diseases. Duration 1 – 3 days RHD is caused by caliciviruses, small, non-enveloped, single-stranded RNA viruses. Rabbit haemorrhagic disease virus (RHDV) is closely related to the European brown hare syndrome virus, which causes a similar disease in hares (Lepus spp.). There are several genetically and serologically different variants of RHDV. Until 2010, six different 13.1.39 Rabies Virus genotypes were known which cross-react serologically. These are called “classic” RHDV or RHDVa. A new serotype, called RHDV2 or RHDVb, was first detected in France Rabies virus (RABV) belongs to the genus Lyssavirus of the family Rhabdoviridae and in 2010 and has since spread throughout Europe and other parts of the world. The is globally distributed. In Germany, rabies is a notifiable epizootic disease. After disease caused by RHDV2 is similar to that of classic RHDV strains but is associated intensive control measures, Germany has been considered free of classical rabies with a slightly lower (but extremely variable) mortality rate. RHDV2 can also infect some (RABV) since 2008. hare species and, unlike RHDVa, also infects very young rabbits. When travelling, some countries demand proof of the antibody titre. From January 2015 to June 2017, only 0.6% of Laboklin samples were RHDV/RHDVa Rabies – Antibodies* positive, but 37.4% were RHDV2 positive. RHDV/RHDVa was only detected in samples Material S 1 ml from Germany and the Netherlands, while RHDV2 was detected in animals from Method FAVN Germany, Great Britain, Luxembourg, the Netherlands, Spain, Switzerland, Poland, Species Dog, cat Belgium, Austria, Sweden and Finland. Duration 1 – 2 weeks RHDV/RHDVa and RHDV2 are mainly transmitted orally. Contaminated herbage can play a role here. Insects also act as mechanical vectors. Note Control of vaccination titres, also for export. Infected animals often show general clinical symptoms such as anorexia and lethargy, Please request special form if the test results are required for export. but also neurological symptoms such as opisthotonus, excitement, ataxia or paralysis. Conjunctivitis and respiratory symptoms such as dyspnoea and nasal discharge (possibly bloody) are also frequently observed. In some cases, an increased tendency 13.1.40 Ranaviruses to bleed can be observed. The chronic form of RHD only occurs in a small number of animals which then develop jaundice. Ranaviruses are enveloped double-stranded DNA viruses and belong to the family Hepatomegaly and splenomegaly are the most common pathologies. Histologically, Iridoviridae. They are found worldwide and have a very wide host range infecting acute necrotising hepatitis can be detected in affected animals. Bleeding and blood different animal species and even classes. Transmission is by direct contact, stasis in various organs are frequently observed. Affected rabbits often die within a few environmental contamination or cannibalism (or eating infected animals). days. In amphibians, ranaviruses are increasingly detected and can in these trigger systemic In addition to the clinical examination and histopathology, RHD is mainly diagnosed by disease and mass death. Clinically, distinctions are made between a haemorrhagic virus detection using realtime PCR. Due to the genetic differences between the RHDV form and a skin form. strains, both RHDV/RHDVa and RHDV2-specific methods must be used. In reptiles, ranaviruses occur especially in turtles, where they are associated with Treatment is not possible. A prophylactic vaccination is recommended. Several stomatitis, rhinitis, pneumonia and liver disease. In lizards, ranaviruses seem to have a vaccines are available. It must be noted that vaccination should take place against both role in skin lesions, stomatitis, granulomatous changes and mass deaths. The clinical RHDV/RHDVa and RHDV2. Currently, RHDV2 cases are being observed in Germany, symptoms of snakes are reported as stomatitis, granulomatous changes and liver but classic RHDV strains still occur. inflammation. Ranavirus is also found in fish. In fish, the infection can extend from clinically inapparent to systemic disease with mass mortality. 170 171 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS Ranaviruses – Pathogen Detection 13.1.42 Rotaviruses Material Tortoise/turtle: swab without medium (pharynx, cloaca), tissue Group A rotaviruses are one of the most important pathogens of nosocomial (liver, tongue, kidney, intestine, possibly skin), possibly EB gastroenteritis in veterinary and human medicine. In Germany, human rotavirus infections (box turtles) are among the most common gastrointestinal diarrhoeal diseases and are a notifiable Lizard: swab without medium (pharynx, cloaca), tissue (above all infection. Rotaviruses are non-enveloped and therefore very environmentally stable skin, liver) viruses. Rotaviruses are transmitted both via the faecal-oral route and airborne. The Snake: swab without medium (pharynx), tissue (above all liver) destruction of enterocytes and electrolyte imbalances lead to diarrhoea and dehydration. Amphibians: biopsy (toe clips, tail clips), blood, tissue (above all Detection is done from faeces; the virus antigen is detected by ELISA. liver, kidney), perhaps swab without medium (skin) Fish: biopsy (gills), blood, tissue (above all liver, kidney), perhaps swab without medium (skin) Rotavirus – Antigen Method PCR Material Faeces Species Reptiles, amphibians, fish Method EIA Duration 1 – 3 days Species Dog, cat, horse, cattle Duration 1 day Note A faecal sample of at least 2 g is required. 13.1.41 Reoviruses Birds 13.1.41 Sacbrood Virus Reoviruses are non-enveloped double-stranded RNA viruses that are regularly found in various bird species. They can cause inapparent infections but are also associated Sacbrood virus is an RNA virus of the picornavirus family. This virus only affects bee with various clinical changes. Liver and intestine are often affected. Respiratory infection brood; infected adult animals do not show any symptoms. Transmission occurs through also occurs. Especially in Old World parrots, lethal infections can be seen. the bees that take up the virus when removing dead larvae and afterwards excrete it again through the hypopharyngeal glands when feeding. The virus can survive winter dormancy Reptiles in the salivary glands. Reoviruses are often found in snakes and lizards, but occasionally also in turtles. In snakes and lizards, they are associated with respiratory symptoms, particularly pneumo- Infected larvae die shortly before pupation and become small, fluid-filled “sacs” which nia. However, they could also be involved in skin lesions (papillomatous changes). In later dry out and change to scab. The brood pattern shows sunken caps. Sacbrood is turtles, reoviruses are associated with respiratory symptoms and stomatitis. considered a so-called secondary infection, since the disease usually only takes on a severe course if the colony has been weakened by a primary infection, such as varroosis. Reoviruses – Pathogen Detection For the therapy of the swarm, affected combs can be removed and melted. Transmission Material Birds: swab without medium (cloaca), faeces, tissue (intestine, occurs through the bees themselves or through the beekeeper. liver, heart, kidney, lung) Reptiles: swab without medium (pharynx, cloaca), lung lavage, Sacbrood Virus – Pathogen Detection tissue (lung, intestine, tortoise/turtle: tongue) Material Bee larvae Method PCR Method PCR Species Birds, reptiles Species Bees Duration 2 – 4 days Duration 1 – 3 days 172 173 2019/20 Infectious Diseases / Viruses 2019/20 Infectious Diseases / Viruses LABORATORY FOR CLINICAL DIAGNOSTICS 13.1.44 Sendai Virus brain stem and sometimes also the spinal cord. The disease usually begins acutely to peracutely with a highly elevated body temperature (up to over 41 °C) and a further Sendai virus, also known as murine parainfluenza virus 1, causes infections in rapidly progressive course. Changes in behaviour, from being apathetic to overexcited rabbits, guinea pigs, hamsters, rats and mice as well as in humans. An introduction or aggressive, gait disorders up to tetraparesis/tetraplegia and seizures can occur. into a population leads to severe respiratory symptoms (focally ulcerative/necrotising Various brain nerve deficits are observed, e.g. facioplegia, strabismus, nystagmus, rhinitis/tracheitis, pneumonia and pleuritis) and mortality rates of up to 100%, especially miosis, missing menace reflex. Hyperalgesia in the head and neck area as well as a in mice. If the infection persists in a population, the course of infection is milder or generally increased painfulness are characteristic. A large part of the disorders ends subclinical. Once the infection is overcome, antibodies are detectable throughout life. lethally or by euthanasia within one week. Recently, there have been more and more literature reports on dogs with a chronic course of the disease that have survived. Sometimes small neurological symptoms remained, sometimes the dogs were fully Sendai Virus – Antibodies* recovered. Material S 0.5 ml Method IFAT Diagnosis should be confirmed serologically by antibody detection using ELISA. Species Rabbit, guinea pig, rat, mouse, hamster However, it must be taken into account that the antibodies could be the result of a Duration 1 week previous subclinical infection. Antibodies may also appear in the liquor within the first week after infection and can be detected by ELISA. In the peracute form, PCR can be used to try to detect the virus in the liquor. Due to 13.1.45 Sunshine Virus the very rapid virus elimination from the brain, however, this is only possible in the early phase of infection. Virus detection by PCR from a collected tick is possible and Sunshine virus is a novel paramyxovirus (PMV), which was first demonstrated in especially useful if a person is affected by a tick. pythons in 2012 in Australia. Sunshine virus is only distantly related to Ferlavirus (formerly known as reptile PMV or snake PMV). It has been demonstrated in animals Tick-borne Encephalitis Virus (TBE Virus) – Pathogen Detection with respiratory and/or neurological symptoms, but can occasionally be detected even in clinically healthy animals. Initial studies show that this virus may also be present in Material CSF 0.2 ml, S 0.2 ml, tick European pythons. Method Realtime PCR Species Dog, horse and others Duration 1 – 3 days Sunshine Virus – Pathogen Detection Note Detection from serum (before seroconversion) or cerebrospinal fluid Material Swab without medium (pharynx, cloaca), tissue (lung, brain) is only possible in the early phase of the infection. Method PCR Species Python Duration 1 – 3 days Tick-borne Encephalitis Virus (TBE Virus) – Antibodies Material S, EP, HP, CSF 0.5 ml Method ELISA 13.1.46 Tick-Borne Encephalitis Virus (TBEV) Species Dog, cat, horse Duration 3 days Tick-borne encephalitis (TBE) is caused by an arbovirus. Arboviruses are an Note Performing the analysis is advisable if animals have been to inhomogeneous group of viruses whose common feature is the transmission by blood- endemic areas and show neurological symptoms. sucking arthropods. Like the West Nile virus, the TBE virus (TBEV) belongs to the genus Flavivirus and is transmitted by ticks. In dogs, the disease was first described in 1972. Seroepidemiological studies conducted since then have shown that dogs have relatively frequent contact with TBEV (up to 30% in certain areas) without contracting the disease. If the disease is contracted, the symptoms in dogs are a multifocal occurrence involving the cerebrum, 174 175 2019/20 Infectious Diseases / Viruses, Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS 13.1.47 West Nile Virus Actinobacillus pleuropneumoniae (APP) – Antibodies* Material S, HP 0.5 ml The virus is transmitted by blood-sucking insects. Susceptible dead-end hosts are Method ELISA horses and humans; birds can become infected and serve as the disease reservoir, Species Pig thus possibly transferring the virus over long distances. However, some avian species, Duration 5 days such as crows, are very sensitive and quickly die from the infection. Infected horses show symptoms of encephalitis, but also ataxia, tremors and seizures as well as Actinomyces Ø see Chapter 14.4 paralysis. The mortality rate in infected horses is about 30%. In birds and horses, an infection with West Nile virus is a notifiable disease in Germany. 13.2.2 Anaplasma West Nile Virus – Antibodies Based on genetic analyses, the former species Ehrlichia phagocytophila, Ehrlichia equi Material S 0.5 ml and the causative agent of human granulocytic ehrlichiosis (HGE) were unified in the Method ELISA new species Anaplasma phagocytophilum. In addition, infection with Anaplasma platys, Species Birds, horse, pig the causative agent of infectious canine cyclic thrombocytopenia, plays an increasingly Duration 3 – 5 days important role in Europe, too. 13.2 Bacteria Anaplasma phagocytophilum Anaplasma phagocytophilum is a gram-negative, obligate intracellular bacterium, which particularly infects neutrophil granulocytes and forms, when multiplying within the 13.2.1 Actinobacillus pleuropneumoniae (APP) granulocytes, typical inclusion bodies, so-called morulae. In Europe, the main vector is Ixodes ricinus. Deer, mice and other rodents are reservoir hosts. Actinobacillus pleuropneumoniae is a gram-negative, rod-shaped bacillus. It produces The clinical symptoms are similar to those of ehrlichiosis, but here, more exotoxines that can destroy erythrocytes and lung macrophages. The clinical picture of thrombocytopenia can be observed, i.a. due to the formation of anti-thrombocyte APP infection is characterised by severe respiratory symptoms and significant impair- antibodies. ment of the general condition (body temperature may rise up to 42° C). In intensive pig The Anaplasma infection can by asymptomatic and cause non-specific symptoms farming, pleuropneumonia caused by A. pleuropneumoniae is one of the most impor- (fever, inappetence, apathy) or severe symptoms (CNS disorders). In dogs, orthopaedic tant infectious diseases. In peracute courses of the disease, it can lead to the death of problems (myositis, joint swellings, lameness) are often associated with Anaplasma the animals within 24 hours. infections. In horses, fever, apathy, limb oedema and reluctance to move are initially dominant. Actinobacillus pleuropneumoniae (APP) – Pathogen Detection Horses older than 4 years show more obvious symptoms than younger animals. Once Material (1) Swab with medium, tissue (lung, tonsils) the infection is overcome, a resilient immunity is acquired for about 2 years. (2) Swab without medium (nose), nasal flush, tissue In ruminants, Anaplasma phagocytophilum can cause tick-bite fever. Most infections Method (1) Culture progress subclinically, but fever and productivity loss or abortions are also possible. (2) PCR* Severe cases occur when non-immune animals are introduced into endemically Species Pig contaminated areas. Duration (1) 1 – 3 days (2) 7 – 14 days Anaplasma phagocytophilum – Pathogen Detection Material EB 0.2 ml, bone marrow, synovia, CSF 0.2 ml, tick Method Realtime PCR Species Dog, cat, horse, cattle and others Duration 1 – 3 days 176 177 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Note PCR is positive in blood smears 6 to 8 days before and 3 days after 13.2.3 Bartonella henselae the formation of morulae. Similar to an infection with Ehrlichia canis, persistent infections in Bartonella are gram-negative, facultative intracellular bacteria which are transmitted by the bone marrow, spleen and liver are considered for Anaplasma fleas and ticks. Bartonella henselae is mostly known as the causative agent of “cat- phagocytophilum. Thus, a negative PCR result does not completely scratch disease” in humans. Infections in cats are predominantly subclinical. Fever, rule out an infection. muscular pain, local lymphadenopathy and, rarely, also neurological symptoms can occur, which usually disappear again after a few days. Recently, the involvement of Bartonella Anaplasma phagocytophilum – Antibodies henselae in gingivitis and stomatitis in cats has been discussed more frequently. Pathogen detection and antibody detection often do not match and a definitive diagnosis is linked to Material S, EP, HP 0.5 ml the detection of the pathogen. A negative PCR result does not exclude an infection with B. Method IFAT; dog: ELISA henselae and should be repeated in case of clinical suspicion. Species Dog, cat, horse, cattle Duration 1 day Bartonella henselae – Pathogen Detection Note Dog: after consultation, determination can also be done by IFAT. Material EB 0.2 ml, CSF 0.2 ml, swab without medium (oral cavity), flea Method Realtime PCR Species Cat Anaplasma platys Duration 1 – 3 days Anaplasma platys (formerly Ehrlichia platys) is an obligate intracellular, gram- negative bacterium in dogs which multiplies in thrombocytes and leads to cyclic thrombocytopenia and bacteraemia with intervals of approximately 14 days. The Bartonella henselae – Antibodies disease is called infectious canine cyclic thrombocytopenia. Descriptions of this Material S, EP, HP 0.5 ml species of Anaplasma come from overseas, but the pathogen is also detectable in the Method IFAT Southern Mediterranean (North Africa, southern Portugal, Andalusia, Sicily, southern Species Dog, cat Italy, southern Greece). It is transmitted through ticks (Rhipicephalus sanguineus). After Duration 1 day the initial infection, there is a decrease in the platelet count within 7 days p.i.; the lowest Note Antibodies can usually be detected from the second week p.i. values are reached between days 14 and 24 p.i. onwards. Seroprevalence is particularly high in cats with flea Basophil inclusions (morulae) in the thrombocytes can particularly be detected 7 – 10 infestation and is not indicative of a clinical condition. The direct days p.i. The phase of bacteraemia extends approximately over a period of 4 – 14 days detection of the pathogen by PCR is preferable. p.i., followed by a phase in which the pathogen cannot be detected in the peripheral blood. Subsequently, these phases alternate cyclically depending on the platelet count. In the bacteraemic phase, the pathogen can be detected in blood samples by means of PCR. 13.2.4 Bordetella bronchiseptica Bordetella are small gram-negative bacilli which can move by means of flagella. B. Anaplasma platys – Pathogen Detection bronchiseptica usually only survives for a rather short period of time outside the respira- Material EB 0.2 ml, tick tory tract. Transmission takes place by direct contact or via aerosols. Method Realtime PCR Because of its toxins, B. bronchiseptica particularly damages the cilia-bearing cells of Species Dog the respiratory mucosa and it can persist in the respiratory tract for up to three months. Duration 1 – 3 days B. bronchiseptica is not host-specific and can be transmitted from one animal species (e.g. dog) to another (e.g. cat) and also to humans (zoonosis!). In dogs, Bordetella are known as a component of kennel cough and they are also responsible for respiratory tract diseases in cats, although coughing is not a characte- ristic symptom in cats. Typical symptoms are fever, sneezing, nasal discharge, swelling of the submandibular lymph nodes and intensified breath sounds. Usually, only mild symptoms occur which disappear again after about 10 days. However, life-threatening bronchopneumonia can develop in young kittens. 178 179 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Bordetella bronchiseptica – Pathogen Detection Infections and diseases in cats and cattle are reported more and more often. Furthermore, Lyme disease is classified as an emerging bacterial zoonosis. Material (1) Swab imperatively with medium (Amies) (nose, pharynx), Grazing animals are often used for blood meals by borrelia-infected ticks. Clinical bronchoalveolar lavage diseases appear as well as seropositive animals without any clinical signs, with the (2) Swab without medium (nose, pharynx), bronchoalveolar lavage evaluation often being difficult. Method (1) Culture (MALDI-TOF) In horses, a variety of symptoms are associated with borrelia: reduced performance, (2) Realtime PCR lameness, changes in the skin, eyes or heart up to neurological deficits and abortions. Species Dog, cat, rabbit, cattle, sheep, goat, pig, others However, there is still controversy as to whether the infection in horses leads to any Duration (1) 2 – 3 days clinical symptoms at all. (2) 1 – 3 days Note When requesting a bacteriological examination, please clearly Lyme disease in cattle is associated with lameness, weight loss and abortion. indicate on the submission form that Bordetella bronchiseptica Pathogen isolation from clinical material is sometimes successful (Borrelia burgdorferi should be tested, as special culture media are required. sensu stricto, Borrelia afzelii). Seroconversions have been shown as well as the response to tetracycline therapy. 13.2.5 Borrelia Borrelia – Pathogen Detection Material Tick, biopsy (skin), synovia, CSF 0.2 ml Borrelia are bacteria which belong to the spirochaetes. Spirochaetes are characterised Method Realtime PCR by contractile axial filaments which are located under a multi-layered outer membrane Species Dog, cat, horse, cattle, sheep, goat, other species and that give the spirochaetes their typical spiral shape as well as their motility. Borrelia Duration 1 – 3 days species which are discussed in connection with Lyme borreliosis in dogs are included in the group Borrelia burgdorferi sensu lato, which currently comprises 19 different Note The diagnostic value of a PCR is limited by the selection of the Borrelia species. appropriate material and the concentration of pathogens. During a chronic infection, pathogen spread can be suspected in many Borrelia are transmitted by vectors (ticks or lice) and except for B. recurrens and B. sites, but the concentration of pathogen DNA can be very low and dutonii they all have a reservoir among wild animals. therefore the PCR produces a negative result. While a positive PCR The main mode of transmission is a bite of the tick Ixodes ricinus (European castor is proof of infection, a negative PCR does not exclude an infection. bean tick). The bacteria are located in the intestine of the tick, are activated by the blood meal and migrate to the salivary glands. It then takes up to 24 hours until Borrelia – Antibodies (IgM+IgG) transmission via the saliva takes place. If the tick is properly removed within this period, Material S, EP, HP 0.5 ml the risk of infection can be greatly reduced. Method IFAT; dog: ELISA In contrast to humans, the clinical symptoms of Lyme borreliosis (Lyme disease) in Species Dog, cat, horse, cattle dogs are rather unspecific and can easily be overlooked. In dogs, there is rarely an Duration 1 day erythema migrans. Fatigue, loss of performance, possibly fever and, after a symptom- free phase of several weeks, reluctance to move, alternating lameness, emaciation, Note • Positive IgG antibody titres are found in dogs after about 3 – 6 vomiting and oedema occur. Occasionally, neurological deficits are also observed. A weeks, positive IgM antibodies 3 – 4 days after pathogen contact. serious complication is the development of glomerulonephritis with subsequent kidney The distinction between IgM and IgG is used to distinguish an failure due to the deposition of immune complexes. acute infection from a pathogen contact that occurred a long time The main vector, Ixodes ricinus, occurs throughout Germany but can be found more ago. frequently in certain areas. In such areas, it is therefore recommended to regularly • After consultation, detection in dogs can also be done using IFAT. check for any infestation of the dog with ticks and to have a Lyme disease test carried out if the symptoms mentioned above occur. 180 181 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Borrelia – Blot 13.2.7 Brucella Material S, HP 0.5 ml The causative agent of brucellosis are gram-negative, aerobic bacilli of the genus Method Line lmmunoassay/western blot Brucella. Brucellosis in cattle, pigs, sheep and goats is notifiable upon suspicion. (specific antibody detection) The disease occurs in both animals and humans. Several Brucella species are known, Species Dog, horse including B. canis (canine brucellosis), B. abortus (bovine brucellosis), B. melitensis Duration 1 – 3 days (ovine and caprine brucellosis), B. ovis (brucellosis in rams, infectious epididymitis) and Note The immunoblot is used to clarify B. suis (porcine brucellosis). Host-specificity of Brucella species is only limited. numerous questionable antibody titres and to distinguish between Brucella canis is transmitted genitally or via the oral route by latently infected animals. vaccination and infection antibodies. After 2 to 4 weeks, bacteraemia develops. In pregnant female dogs, there may be The blot also detects antibodies abortions in the last trimester of gestation or weak puppies are born. Male dogs suffer against the VlsE protein and from inflammation of the testicles and epididymis and can become infertile. The main synthetic C6 peptide. VlsE (variable symptoms of brucellosis in ruminants are abortions, birth of weak animals, inflammation major protein-like sequence, of the testicles and epididymis, and infertility. In humans, the infection leads to fever, expressed) and its subunit C6 are fatigue, night sweats, headaches and feelings of cold. The occurrence of cases in hu- highly immunogenic surface mans is always related to the disease being present in domestic or wild animals. antigens that express Borrelia during active reproduction. There is no recombination of the VslE molecule in vitro or in the Borrelia resi- Brucella canis – Pathogen Detection ding in the tick, the detection of antibodies against the VslE molecu- le therefore indicates an infection that has previously occurred. Material Swab without medium (cervix, prepuce), EB 0.2 ml, sperm, urine, The blot can be carried out at the earliest from the 3rd week after (faeces, milk), tissue (abortion material) infection onwards. Method Realtime PCR Species Dog Duration 1 – 3 days 13.2.6 Brachyspira Brucella canis – Antibodies Brachyspira are gram-negative anaerobic bacteria which, however, have a certain Material S 1 ml tolerance to oxygen. Reproduction takes place in the goblet cells of the large intestine Method (1) IFAT where Brachyspira can persist after surviving infection (intermittent shedding!). Pig (2) MAT dysentery, which is caused by B. hyodysenteriae, is a highly contagious multifactorial Species Dog diarrhoeal disorder that leads to high economic losses in pig production worldwide. The Duration 1 day main sources of infection are infected pigs without clinical symptoms and rodents as reservoir hosts. B. pilosicoli causes a milder disease, spirochete diarrhoea in pigs, Note Agglutination tests are usually demanded as certificate for transport which usually occurs directly after weaning. in non-European countries. Brucella abortus and Brucella mellitensis – Antibodies Brachyspira hyodysenteriae/pilosicoli – Pathogen Detection Material S, HP 0.5 ml Material Faeces, tissue (large intestine) Method ELISA Method Realtime PCR Species Cattle, sheep, new world camelids Species Pig Duration 3 days Duration 1 – 3 days Note Differentiation between B. hyodysenteriae and B. pilosicoli is done by PCR. 182 183 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Brucella suis – Antibodies In dogs and cats, C. jejuni can often be isolated from healthy animals, but can cause diarrhoea especially in young animals. This diarrhoea is often self-limiting. Feeding a Material S, HP 0.5 ml barf diet presents a risk of infection. Method ELISA Infections in humans, most commonly with C. jejuni, are usually food-associated. Species Pig Campylobacter is one of the most frequent causative agents of bacterial diarrhoeal Duration 3 days diseases in humans (especially due to insufficiently heated or contaminated poultry meat or poultry products, except for eggs). Further sources of infection include unpasteurised milk as well as raw minced meat. Rare complications which can occur 13.2.8 Burkholderia mallei are Guillain-Barré syndrome (polyradiculitis) and reactive arthritis. Campylobacter of the species C. jejuni, C. coli, C. lani and C. upsalensis are collectively Glanders is a disease of Equidae caused by Burkholderia mallei, which also has called thermophilic Campylobacter. In Germany, campylobacteriosis (thermophilic zoonotic potential: Apart from humans, wildcats (zoos!), camels, bears, wolves and Campylobacter) is reportable upon diagnosis in dogs, cats, ruminants and poultry. dogs are susceptible as well. Cattle, sheep and pig are resistant. The course of the disease is either acute (especially in donkeys and mules) with high fever and respiratory Campylobacter – Pathogen Detection symptoms and death after a few days. Or it is rather chronic, particularly in horses, with nodules and ulcerations on the skin, the mucous membrane and the inner organs. Material Faeces, swab with medium (intestine, cloaca) Chronically and subclinically infected animals represent a dangerous source of Method (1) Culture (MALDI-TOF) infection. All secretions of the respiratory tract and the skin are infectious; the incubation (2) Realtime PCR (only detection of Campylobacter jejuni) period ranges from a few days to many months. Species No limitations known In Europe, glanders is considered eradicated, but it does occur in different Asian, Duration (1) 2 – 3 days African and South American countries (export-relevant test). (2) 1 – 3 days In Germany, there is an obligation to inform the authorities! Note • Culture: faecal sample with a diameter of at least 2 cm, otherwise use swab with medium; for PCR: swab without medium. Burkholderia mallei (glanders) – Antibodies* • A combined cultural detection of Campylobacter and Yersinia is also available. Material S 1 ml • Resistances are common; therapy should therefore only be Method CFT carried out after an antibiogram has been performed. Preparation Species Equidae of an antibiogram is only possible after cultural examination. Duration 5 days 13.2.10 Candidatus Neoehrlichia mikurensis 13.2.9 Campylobacter Officially named in 2004, candidatus Neoehrlichia mikurensis is an obligate intracellular, Several Campylobacter species could be detected in mammals, birds and also gram-negative bacterium. The pathogen is characterised by endotheliotropism but has in humans. Some species are part of the normal gastrointestinal microbiota or its not been cultivated in vitro so far and thus could not be completely described yet. pathogenicity is still unclear. Cand. N. mikurensis was first found in common rats on the Japanese island of Mikura. In cattle, C. fetus subsp. veneralis causes epidemic abortions and fertility disorders It is assumed that small mammals, such as mice and rats, serve as reservoir; transmis- (reportable upon suspicion), C. jejuni can lead to diarrhoea or mastitis. In sheep, C. sion most likely occurs through ticks. In recent years, Cand. N. mikurensis has been fetus subsp. fetus is known as pathogen of the enzootic Campylobacter abortion; detected in about 2 to 25% of Ixodes ricinus ticks in Germany. occasional abortions caused by C. jejuni have also been described. The importance of Campylobacter infections in birds lies in the contamination of carcasses and the risk Since 2007, this pathogen has been associated with diseases in humans. Especially of food poisoning associated with it. Most frequently, birds are infected with C. jejuni. the elderly and immunocompromised people have been affected by the so-called Diarrhoea or hepatitis are rare. neoehrlichiosis, including two patients from Germany. The symptoms are non-specific, most often high fever and headaches as well as muscular pain and joint pains were 184 185 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS seen. The occurrence of vascular complications, like deep vein thromboses, pulmonary can last for 8 weeks or more. Other acute symptoms include slight rhinitis and fever. embolism and arterial aneurysms, was most noticeable. Laboratory findings particularly Animals between 5 weeks and 9 months of age are most affected, though conjunctivitis indicate an increased level of C-reactive protein, leukocytosis with neutrophilia and neonatalis has also been described. In that case, kittens already suffer a severe anaemia. conjunctivitis when opening their eyes, often due to a Chlamydia infection acquired intrapartum. Transmission occurs directly through conjunctival secretions. Persistent In dogs, so far only one single case has been reported in which this bacterium could infections are possible, and in some animals also respiratory symptoms may last for be isolated. It was an eight-year old female Irish Setter after ovariohysterectomy and several weeks. When the immune system is weakened, the infection can be reactivated. mastectomy. Postoperatively, she was lethargic and developed profuse subcutaneous bleeding (Diniz et al. 2011). Birds In our birds, infections with Chlamydia are of particular significance. Infection rates of 10 Candidatus Neoehrlichia mikurensis – Pathogen Detection to 40% may be prevalent in aviculture. As many birds have a carrier status, the disease can “suddenly” become clinically apparent under stress. Symptoms in birds are Material EB 0.2 ml, tick, tissue (e.g. spleen, kidney, liver) manifold and extremely non-specific. Ruffled feathers, apathy and lack of appetite must Method Realtime PCR be mentioned. Basically, every “sick bird” could have a Chlamydia infection. Respiratory Species Dog, tick symptoms with or without conjunctivitis are often seen, but central nervous disorders Duration 1 – 3 days are also possible. The extent of clinical signs largely depends on the animals’ condition; the type of symptoms also varies from one bird species to another. Sudden deaths without prior illness might happen. It is therefore not possible to make a diagnosis 13.2.11 Chlamydia based on clinical signs. To make a reliable diagnosis, the identification of the pathogen is always necessary. C. psittaci is a zoonotic agent. Infections in humans are normally Chlamydia are obligate intracellular, gram-negative pathogens. They cannot replicate airborne, resulting in a flu-like disease. In Germany, it is a notifiable disease. on their own, but need the enzyme activity of a suitable host cell. Reptiles and amphibians Chlamydia relevant in veterinary medicine belong to the family Chlamydiaceae. Some Various species of Chlamydia are regularly detected in reptiles and amphibians. In years ago, this family was split into the two genera Chlamydia and Chlamydophila. reptiles, they have been associated with granulomatous changes in different tissues as However, based on recent genetic analyses, this classification is no longer considered well as with pneumonia, enteritis, hepatitis and myocarditis. In amphibians, they were justified. Because of this, the single term Chlamydia is used here. found in cases of systemic disease. Farm animals Dog In Germany, chlamydiosis in cattle, sheep and goat, as well as in birds (see above) is a In literature, only little data is available on Chlamydia infections in dogs. Yet in notifiable disease. general, its appearance must be expected in Europe. Respiratory signs up to bronchopneumonia seem to be dominating here. At the onset of the disease, only progressive loss of condition may appear. High fever can develop. When the Chlamydia – Pathogen Detection disease progresses, central nervous disorders are possible. Other manifestations Material Dog, cat and others: swab without medium (eye, pharynx, cervix, of chlamydiosis in dogs are conjunctivitis and keratitis. Its involvement in keratitis prepuce), abortion material superficialis chronica in German Shepherd dogs is being discussed. Birds: triple/3-fold swab without medium (eye AND pharynx AND cloaca), tissue (liver, spleen, lung), if need be faeces Cat Reptiles: swab without medium (pharynx), lung lavage, tissue Originally identified as the causative agent of “feline pneumonitis”, C. felis is rather (lesions, lung, liver, spleen, intestine, heart) associated with conjunctivitis in cats nowadays. The cardinal symptom is serous Farm animals: swab without medium (eye, nose, cervix), lavage conjunctivitis which begins unilaterally and then spreads to the other eye. Especially sample, milk, faeces, tissue (lung, liver), abortion material when there is a secondary bacterial infection, the discharge can become mucopurulent. Amphibians: swab without medium (pharynx), tissue (lesions, lung, Chemosis and blepharospasm may also be present. In severe cases, follicular liver, spleen, intestine, heart) hyperplasia develops or even keratoconjunctivitis with corneal ulcerations. Conjunctivitis 186 187 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Method Realtime PCR Note Examination is specifically indicated if colitis-like symptoms occur. Species All A faecal sample of 2 cm of diameter or larger is required. Duration 1 – 3 days Note Pathogen detection identifies all Chlamydia of the family Clostridium perfringens Enterotoxin Chlamydiaceae. Material Faeces Method ELISA Chlamydia – Antibodies Species No limitations known Material S, EP, HP 0.5 ml Duration 2 days Method IFAT Note Determination is particularly indicated in case of colitis. Species Dog, cat, birds, horse, cattle In carnivores, Clostridium perfringens enterotoxin can cause Duration 1 day diarrhoea and vomiting of varying severity; enterotoxaemia is rare. Note Serology can determine whether an infection is or was present. Toxin formation is induced by antibiotic administration, stress, co- However, proof of active shedding can only be provided by infections or especially by an unbalanced diet rich in proteins and pathogen detection. connective tissue. Detection of further Clostridia on request. 13.2.12 Clostridia Clostridia are obligate anaerobic, gram-positive, spore-forming bacilli. 13.2.13 Coxiella burnetii Pathogenic Clostridia cause infectious and intoxication diseases; the latter through enterotoxins and neurotoxins. Coxiella burnetii is an obligate intracellular, gram-negative bacterium and the pathogen that causes Q-fever. It is highly infectious, only a small amount is needed to establish an infection. Clostridium botulinum Neurotoxin – Antibodies* Coxiella burnetii is spread worldwide and pathogen to a wide range of species, e.g. Material S 1 ml ruminants, dogs, cats, rodents, birds and humans (zoonotic!). An infection in humans is Method ELISA often subclinical but clinically unspecific severe influenza-like symptoms can occur. Fur- Species Horse, cattle, other farm animals on request thermore, chronic forms with endocarditis, hepatitis or CNS involvement are described. Duration 10 days Especially persons who are in contact with ruminants (e.g. vets, farmers, butchers) are affected. Note Botulism is regarded as pure intoxication, in which only the In ruminants, Coxiella burnetii replicates in the female genital tract and in the utter. It toxin is absorbed, reabsorbed via the intestine and spread is intermittently or persistently secreted by uterus secretion, amnion fluid and abort haematogenously. If, in exceptional cases, botulism progresses material, but also by urine, faeces or milk. During replication, small spore-like forms as a toxin infection, the toxins are formed in the intestine (visceral are produced, which can survive in the environment for a very long time. Transmission botulism) or in wounds (wound botulism). The absorption of occurs mostly by inhalation of pathogenic dust, but also by direct contact with infected botulinum toxin leads to paralysis of the motor nerves. animals (e.g. during obstetric assistance). Ticks have also been found to be vectors of Coxiella burnetii, whereby the tick faeces are infectious. Clostridium difficile Toxin A and B As in humans, the infection with Coxiella burnetii in animals is mostly subclinical. Clini- Material Faeces cal symptoms in animals are metritis, retained placenta, embryonic death, late abor- Method ELISA tions and stillbirths with subsequent infertility or birth of small, weak calves. Species No limitations known The finding of Coxiella burnetii in cattle, sheep and goat is notifiable in most countries! Duration 2 days 188 189 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Coxiella burnetii – Pathogen Detection Ehrlichia canis – Pathogen Detection Material Swab without medium (genital tract), abortion material, milk, faeces, Material EB 0.2 ml, bone marrow, tick urine Method Realtime PCR Method Realtime PCR Species Dog, cat Species Mainly ruminants, but also other species Duration 1 – 3 days Duration 1 – 3 days Note As this is a zoonotic disease, the education of the animal owner and Ehrlichia canis – Antibodies the practice staff is advisable. Material S, EP, HP 0.5 ml Method IFAT Coxiella burnetii – Antibodies Species Dog, cat Material S, HP 0.5 ml Duration 1 day Method IFAT Species Dog, cat, horse, cattle, sheep ESBL Ø see Chapter 14.4 Duration 1 day Dermatophilus congolensis Ø see Chapter 14.4 13.2.15 Francisella tularensis E. coli, eae/enteropathogenic Ø see Chapter 16.1.2 Francisella (F.) tularensis is a gram-negative, pleomorphic, non-motile, aerobically growing, rod-shaped bacterium that is very resistant especially in lower environmental temperatures. 13.2.14 Ehrlichia This pathogen is the causative agent of the so-called tularaemia (rabbit fever), which is a zoonosis. Four subspecies have been classified. Two subspecies are of particular clinical relevan- Infections with ehrlichia occur worldwide. Ehrlichia are gram-negative, obligate ce: F. tularensis ssp. tularensis and F. tularensis ssp. holarctica, with F. tularensis ssp. intracellular bacteria belonging to the Rickettsiales and are transmitted by ticks. tularensis naturally occurring in North America only and being responsible for a more Depending on the region, the tick species differ and thus also the species of ehrlichia. aggressive course of the disease. In contrast, F. tularensis ssp. holarctica can be found Whereas in Mediterranean countries and tropical as well as warmer areas (Portugal, the in the entire northern hemisphere. Canary Islands, USA, Asia, Australia), Rhipicephalus sanguineus, the main carrier of E. canis, is prevalent, Ixodes ricinus is found in Central and Northern Europe. However, if It is predominantly hares, rabbits and rodents (mice) that are affected, but also nu- R. sanguineus is introduced into Germany, it can survive in heated rooms. Infection with merous other animal species, including birds, are susceptible to this pathogen. In dogs, Ehrlichia canis still rather presents a “typical” travel-related disease or can be found in cats and sheep, sporadic cases of illness are known. Cats suffer from the disease more imported animals. often than dogs, but overall, the disease is rarely contracted. An infection with E. canis leads to an infection of the monocytes and thus to canine Symptoms of an acute disease are apathy, fever, tachypnoea and swelling of the lymph monocytic ehrlichiosis (CME). The incubation period is about 8 – 20 days, which nodes; most animals die of septicaemia within 2 weeks. Furthermore, in a chronic then changes into an acute phase of 2 – 4 weeks. Clinical symptoms are mostly non- course, emaciation and skin ulcerations occur; in dogs and cats in addition to spleno- specific: fever, anorexia, dyspnoea, anaemia, lymphadenopathy. In rare cases, CNS megaly, hepatomegaly, ulcers on the tongue and jaundice. disorders may occur. In the first 10 – 20 days, thrombocytopenia can be seen, although Here, too, a lethal outcome is possible after 2 – 6 weeks. there is rarely spontaneous bleeding. Subsequently, if untreated, a subclinical stage The modes of transmission include blood-sucking insects like fleas, biting midges, lice, develops, which lasts for months or years, or chronic infections arise, which are often ticks etc., consumption of infected carcass/meat or contaminated water. The infectious accompanied by hypergammaglobulinaemia. E. canis can also infect cats! dose is very low, only a few bacteria are sufficient (exception: infections through the gastrointestinal tract). Humans become infected when frequently exposed to hares, rabbits or wild animals. In Germany, there is an obligation to notify the authorities when F. tularensis ssp. is detected in hares and rabbits! 190 191 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Francisella tularensis – Pathogen Detection PCR accurately detects at least H. pylori, H. felis, H. bizzozeroni, H. hepaticus, H. bilis, H. heilmannii, H. muridarum, H. mustelae, H. Material Tissue (mainly spleen, liver, lung, kidney), lymph node (aspirate), cinaedi, H. canis and Flexispira rappini. swab without medium (pharynx, tonsil) Flexispira rappini is also assigned to the genus Helicobacter and is Method Realtime PCR associated with the occurrence of abortions in sheep. Species (Dog, cat), rabbit, hare, mouse, others Duration 1 – 3 days Note The PCR detects F. tularensis ssp. holarctica. 13.2.17 Histophilus somni Histophilus somni (formerly Haemophilus somnus) is a gram-negative bacillus of the 13.2.16 Helicobacter family Pasteurellaceae. While some strains of H. somni are commensals of the mucosa of the upper respiratory and reproductive tract in cattle, sheep and other ruminants, Helicobacter spp. are helical or curved, gram-negative bacteria. For culture, they pathogenic strains spread systemically and can cause severe diseases such as need especially enriched culture media. The Helicobacter genus comprises at least pneumonia, thrombotic meningoencephalitis, myocarditis, septicaemia, arthritis and 35 species. Some Helicobacter species colonise the gastric mucosa, while others abortions. colonise the intestine and liver of humans or animals. In humans, Helicobacter pylori is correlated with gastritis and stomach ulcers, but it is not normally relevant in animals. Histophilus somni – Pathogen Detection It is an anthroponotic disease – in other words, an infection of an animal with a human pathogen. Material Swab without medium, bronchoalveolar lavage, tissue Pathogenicity of Helicobacter spp. in animals has not yet completely been clarified. Method Realtime PCR Helicobacter mustelae was detected in ferrets with gastritis and stomach ulcers, Species Cattle, sheep, goat Helicobacter heilmanii was found in pigs with stomach ulcers. They also seem to be Duration 2 days associated with gastritis, vomiting and inappetence in dogs and cats. Prevalence is very high, both in healthy as well as in infected animals. Hence, an infection does not always lead to the outbreak of the disease. It is likely that the clinical picture is also 13.2.18 Lawsonia intracellularis influenced by the present Helicobacter spp. (mixed infections are frequent) as well as by the individual host response and environmental factors. Horse Gastric Helicobacter spp. include H. felis, H. bizzozeronii, H. heilmannii, H. salomonis; Especially in older foals, Lawsonia intracellularis causes proliferative enteropathy, the intestinal ones comprise H. canis, H. bilis, H. cinaedi, Flexispira rappini. which is accompanied by significant hypoproteinaemia. The animals also show Transmission occurs by oral-oral route, or possibly also by anal-oral route. Flexispira abdominal pain, reduced general condition and anorexia. Secondarily, oedema and a rappini, which is also assigned to the Helicobacter genus, is associated with the pear-shaped abdomen may occur. occurrence of abortions in sheep. Aborted lambs show multifocal hepatic necroses which are similar to the liver lesions seen in campylobacter infections. Pig Porcine proliferative enteropathy (PPE) in pigs is caused by an infection with the Helicobacter – Pathogen Detection obligate intracellular, gram-negative bacterium Lawsonia intracellularis. Infection occurs via the oral route, the spread mainly through the purchase of infected animals. Often, the Material Vomit, gastric lavage, gastric biopsy, sheep: abortion material infection is subclinical. It is widespread in pig herds, especially among weaners, store pigs Method PCR and fattening pigs. Infected animals suffer from growth disorders and diarrhoea. Species Dog, cat, ferret, sheep Duration 1 – 3 days PPE occurs in four clinically apparent forms: as acute and, if untreated, often fatal porcine haemorrhagic enteropathy (PHE) and as porcine intestinal adenomatosis (PIA), or less Note Positive PCR results from faecal samples do not necessarily indicate often as necrotic enteritis (NE) and as terminal regional ileitis (RI) with thickened and stiff involvement of the stomach (gastritis, stomach ulcer, etc.), as PCR ileum. While PHE mainly affects older fattening pigs and younger breeding pigs, the chro- also detects intestinal Helicobacter spp. For this diagnostic task, nic forms PIA, NE and RI mainly occur in weaners and store pigs. stomach biopsies or vomitus are recommended. 192 193 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Lawsonia intracellularis – Pathogen Detection Horse Leptospira infections, which are spread through the urine of rodents, are usually Material Faeces, tissue (intestine) clinically inapparent in horses. Thus, the seroprevalence in healthy horses is high (up Method Realtime PCR to approx. 75%). The pathogen is ingested with feed or water and leads to rather non- Species Horse (mainly foal), pig specific symptoms in horses, like fever (often intermittent), icterus, inappetence and Duration 1 – 3 days productivity loss. Abortions have been described as well. Transmission of the pathogen between horses does practically not occur. 13.2.19 Leptospira Equine recurrent uveitis (ERU) – It has been proven that an intraocular persistent leptospira infection contributes to the aetiology of ERU. Autoimmune reactions resulting Leptospira are gram-negative bacteria which belong to the spirochete group. They from it lead to a progressive deterioration of the inner structures of the eye and may are very thin, flexible, spiral bacteria with a hook-shaped end. Leptospira can actively even lead to blindness. move by twisting. Within the genus Leptospira interrogans sensu lato, there are various Detection of antibodies (= most sensitive test) or pathogen detection using PCR can pathogenic and saprophytic species which cannot be differentiated morphologically, be carried out from aqueous humour or tissue of the vitreous body. but only serologically or genetically. Since 1989, more than 250 serovars have been described that are currently classified in 24 serogroups. Ruminants Transmission of pathogens occurs directly through the urine or blood of infected Leptospirosis in ruminants can cause economic losses and is a notifiable animal animals or indirectly through inanimate vectors such as contaminated water, feed disease in sheep. It is predominantly cattle kept under extensive grazing conditions and sleeping places or living vectors like rodents. Leptospira best survive in a humid that get infected. In cattle, fever, anorexia, icteric mucous membranes, anaemia with environment at temperatures of 0 – 25 °C. haemoglobinuria and a decline in productivity are dominant. Diarrhoea and mastitis Clinically, leptospirosis in dogs is initially manifested by anorexia, vomiting, dehydration can also occur. Predominant serovars in our own research are L. Icterohaemorrhagiae, and fever. Later, animals are apathetic and often show difficulty breathing. The mucous Saxkoebing and Bratislava. The recently emerged serovar L. Hardjo was not detected in membranes are icteric, anaemia with haemoglobinuria appears and, in some cases, any of the samples we examined. as a complication, disseminated intravascular coagulation (DIC). Toxic degradation products lead to haemorrhagic diathesis and necroses. As a result, acute nephritis with azotaemia can arise. In some cases, hepatitis may also occur, which often has a highly Pig Gravid pigs are especially susceptible to leptospira. The cardinal symptoms are birth of acute course. Leptospira are fetotrophic. weak piglets or abortions. Aborted litters normally show different sizes and degrees of decay of the foetuses as the course of the disease is usually protracted. Dog When testing for antibodies, we look for serovars specific to pigs: L. canicola, L. Grip- According to our own research (see ACVIM Congress 2013, Seattle, USA), there has potyphosa, L. Saxkoebing, L. Bratislava, L. Sejroe, L. Pomona, L. Copenhageni and L. been a shift in the types of serovars over the past years. In dogs, analysed serovars Tarrasovi. include L. Bratislava, L. Australis, L. Autumnalis, L. Icterohaemorrhagiae, L. Pomona, In pigs, there is an obligation to notify the authorities. L. Canicola, L. Saxkoebing, L. Grippotyphosa and L. Sejroe. Cat Leptospira – Pathogen Detection Cats seem to show a natural resistance. However, here too, the number of cases with Material Urine + EB 0.2 ml (bacteraemia only at beginning of the infection), clinical manifestation is increasing. The predominant serovars are L. grippotyphosa and intraocular fluid (horse), tissue (kidney, vitreous body, abortion material) L. Bratislava, followed by L. Icterohaemorrhagiae, L. Sejroe, L. Autumnalis, L. Australis Method Realtime PCR and L. Javanica. Species List Dog, (cat), small mammals, horse, ruminants, pig Duration 1 – 3 days Reptiles Note Test design enables identification of 8 pathogenic serovars of L. In reptiles, leptospira antibodies can be detected quite often. interrogans. 194 195 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Leptospira – Antibodies Listeria – Antibodies Material S, EP, HP 0.5 ml Material S, EP, HP 0.5 ml Method Microagglutination Method IFAT Species Dog, cat, reptiles, horse, ruminants, pig, others on request Species Dog, cat, horse, cattle, sheep, goat Duration 1 day Duration 1 day Note Initially, antibody titres only confirm pathogen contact. Many animals Note Low titres may be unspecific (< 1:80), as there is an antigenic related are seropositive without showing any clinical symptoms. Generally, ness of L. monocytogenes with staphylococci and streptococci. titres > 1:400 or a three- to fourfold titre increase in a paired serum sample at an interval of 14 days are preferred for diagnosis. Peracutely infected animals only show low or even negative antibody 13.2.21 Mannheimia haemolytica titres. Furthermore, if animals have already been treated with antibiotics at a very early stage, the titre often does not increase as Mannheimia haemolytica is a gram-negative, facultative anaerobic bacillus of the genus expected. Mannheimia and the family Pasteurellaceae. It is considered the primary causative For acutely affected animals, direct detection from urine is agent of enzootic bronchopneumonia in cattle and sheep and, moreover, the pathogen recommended. of severe mastitis as well as septicaemia in sheep and goats. In ruminants, however, some of the serotypes described for M. haemolytica are part of the natural microflora of the upper respiratory tract. 13.2.20 Listeria Mannheimia haemolytica – Pathogen Detection Listeriosis can affect many animal species as well as humans. Material Swab without medium, bronchoalveolar lavage, tissue Listeria are relatively small gram-positive rods with a tendency to grow in chains. Within Method Realtime PCR the genus, Listeria monocytogenes has the greatest significance. Listeria ivanovii has Species Ruminants low virulence, but is pathogenic to humans and sheep. The pathogen has also been Duration 2 days isolated from monkeys suffering from meningitis. Listeriosis is predominantly a disease in sheep that contract the disease through the ingestion of spoiled silage. Cattle, chickens, pigs, rabbits and goats are much less 13.2.22 Melissococcus plutonius prone to the disease. Individual cases have been described in horses, dogs and cats. In more than 80% of the cases of ovine listeriosis, the brain is affected and turning The gram-positive bacterium Melissococcus plutonius is the primary pathogen of movements of the head and trunk occur, which are characteristic for the disease. European foulbrood (EFB) in bees. It mainly affects so-called coiled larvae, which then die at 4 – 5 days of age. The larvae are infected through the food and the Other forms are septic listeriosis of newborn or young animals, organ listeriosis (e.g. pathogen multiplies in the gut. Infected brood changes colour and becomes a semi- mastitis) or gestation listeriosis with abortions. liquid mass, which later on dries out to loose scales. Due to the partly sour smell, it is Listeriosis (L. monocytogenes) is a notifiable disease. also referred to as sourbrood. After capping, the caps are sunken and perforated. The symptoms are very similar to those of American foulbrood, a disease notifiable upon suspicion, thus, a precise diagnosis is of great importance. Transmission can either Listeria – Pathogen Detection occur through the bees themselves (drifting, robbing) or by the beekeeper. By forming Material Faeces an artificial swarm, the brood can be separated from the healthy bees and then killed. Method Culture Species Dog, cat, horse, cattle, sheep, goat Duration 3 – 4 days Melissococcus plutonius – Pathogen Detection Material Bee larvae, bees Note • If pathogenic species are detected, an antibiogram will be Method PCR performed additionally (subject to a charge). Species Bees • Detection of listeria is also part of the BARF Profile. Duration 1 – 3 days 196 197 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS 13.2.23 Methicillin-resistant Staphylococci: MRSA/MRSP Normally, an infection already occurs orofaecally in calves through contact with faeces of infected animals, but it can also spread through the colostrum and milk, and intraute- In human medicine, diseases caused by methicillin-resistant Staphylococcus aureus rine infections are possible as well. (MRSA) are known and feared as so-called “nosocomial infections”. These are infec- The incubation period varies greatly and can take several years. The first clinical sym- tions with pathogens that have developed resistance to common antibiotics, often as ptoms often tend to occur when the animals are already older than 2 years. Primary a result of inadequate disinfection measures in hospitals. The pathogens can enter the symptoms are continuous, profound, uncontrollable diarrhoea and progressive weight environment through visitors, personnel, equipment etc. As most of these infections in loss with regular appetite. Paratuberculosis is always lethal. Already prior to the onset of humans are zoonoses, pathogens can also be transmitted to animals as well as vice these symptoms, decreased milk production, reduced fertility, etc. lead to high econo- versa, because of the close contact between humans and animals. This probably also mical losses. Not all infected animals develop clinical symptoms, subclinically infected leads to an increase in MRSA cases in veterinary medicine. carriers also (intermittently) excrete the pathogen. Animals suspected of being infected should be isolated and, in case of a positive result, should soon be eliminated from the In calves, young cattle and pigs, MRSA is detected in about every second animal population or slaughtered. (body) (zoonoses monitoring 2012). In agricultural livestock, MRSA of a certain line are Test results vary depending on the phase of infection, therefore, the use of repeated predominant, so that the term livestock-associated or laMRSA is used. laMRSA mostly sampling is recommended if an infection is suspected! belong to the clonality CC398 and are responsible for 2% of MRSA cases in humans. In Germany, it is a notifiable disease in cattle, sheep and goats! In regions with high livestock density, laMRSA cause up to 10% of human MRSA cases and people with close animal contact, including veterinarians, are particularly at risk. Mycobacterium avium ssp. paratuberculosis – Pathogen Detection In small animals, we detect MRSP, methicillin-resistant Staphylococcus pseudinterme- Material Faeces, tissue (intestine, lymph node), milk dius, far more frequently than MRSA. According to our own research, more than 10% of Method Realtime PCR all isolates of Staphylococcus pseudintermedius (formerly Staphylococcus intermedius Species Cattle, sheep, goat (and others) in dogs) are already afflicted with a multi-resistance gene. Duration 1 – 3 days MRSA/MRSP Mycobacterium avium ssp. paratuberculosis – Antibodies Material Swab (eye, pharynx, nose etc.) Material S, HP, milk 1 ml Method Culture and subsequent MRSA/MRSP differentiation Method ELISA Species Dog, cat, horse, farm animal, others Species Cattle Duration 3 – 4 days Duration 2 – 3 days Note Proof of methicillin resistance by PCR. 13.2.25 Mycoplasma 13.2.24 Mycobacterium avium ssp. Paratuberculosis Mycoplasmas are prokaryotic pathogens that are divided into haemotropic and non- haemotropic mycoplasma. Outside the organism, mycoplasmas are very unstable. Mycobacteria Ø see Chapter 16.1.2 Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of para- 13.2.25.1 Haemotropic Mycoplasma tuberculosis, also called Johne’s disease, a chronic granulomatous enteritis in rumi- nants. The disease is globally distributed. In addition to domesticated ruminants (cattle, Haemotropic mycoplasmas (formerly haemobartonella and eperythrozoon) are globally sheep, goat), also wild ruminants and camelids can be affected. MAP could also be spread, gram-negative bacteria of the family mycoplasmatacea. They attach to the isolated from other animal species, e.g. rabbits, mice, foxes and ferrets. The pathogen surface membrane of erythrocytes and can cause the so-called infectious anaemia. is very stable and can remain infectious in the environment for up to one year. 198 199 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Dog of autoantibodies. Below the normal body temperature (“cold antibodies”), they So far, Mycoplasma haemocanis and Candidatus Mycoplasma haematoparvum agglutinate the blood cells and result in anaemia. Once infected animals go through have been described in dogs. Both strains are found in Europe, especially in the episodes of anaemia time and again. The disease becomes chronic. Older pigs are Mediterranean area. Clinically, the course of the disease is often just chronic and only latently infected and only suffer from another relapse when they are very weak. The asymptomatic. In contrast, acute infections with fever, anorexia, weight loss and pathogen remains in the body throughout life. lethargy are mainly seen in immunocompromised dogs, dogs that had splenectomy or those simultaneously infected with other pathogens. Deaths are also possible. Natural Mycoplasma haemocanis (dog), haemofelis, Candidatus Mycoplasma haemo- infection probably occurs through vectors, particularly the brown dog tick (Rhipicephalus minutum, Candidatus Mycoplasma turicensis (formerly haemobartonella) – sanguineus) is being discussed. Vertical transmission through the placenta and milk is Pathogen Detection also possible, and blood transfusions present a risk of infection as well. Material EB 0.2 ml, tissue (spleen) Cat Method Realtime PCR Species Dog (only M. haemocanis), cat Currently, three different types of haemobartonella with differing pathogenicity have Duration 1 – 3 days been described. In addition to the strain Mycoplasma haemofelis, which is known as Ohio isolate, and the most commonly found California isolate, Candidatus Mycoplasma Note PCR detection should be preferred over microscopic detection as haemominutum, another strain, Candidatus Mycoplasma turicensis, has been known the sensitivity of the microscopic detection is low (around 30%). for some years. The latter was first detected in cats in Switzerland, but seems to occur relatively rarely in Germany. While Mycoplasma haemofelis can cause serious Mycoplasma haemolamae – Pathogen Detection illness even in immunocompetent animals, an infection with Candidatus Mycoplasma haemominutum usually progresses subclinically in healthy animals. Co-infections are Material EB 0.2 ml possible with clinical symptoms typically being more distinct than in mono infections. Method Realtime PCR Natural infection probably occurs through vectors; particularly fleas, but also ticks and Species Llama, alpaca stinging insects are being discussed. Vertical transmission through the placenta and Duration 1 – 3 days milk is also possible. Blood transfusions present a risk of infection, too, as well as direct transmission between animals through bite wounds. Mycoplasma (Eperythrozoon) suis – Pathogen Detection Clinical symptoms in the acute phase are anaemia (haemolytic anaemia as cardinal Material EB 0.2 ml, tissue (spleen) symptom), fever, splenomegaly, general weakness and possibly polypnoea, tachycardia Method Realtime PCR and icterus. The cause of haemolytic anaemia is a haemobartonella-induced damage Species Pig of the erythrocyte membrane. Because of the change in the erythrocyte surface, a Duration 1 – 3 days secondary immune haemolytic anaemia can develop later on; in this case, the direct Coombs test will be positive. The main symptoms of a chronic infection include weight loss and intermittent fever. Studies have shown that a high percentage of the dog and 13.2.25.2 Non-haemotropic Mycoplasma cat population is infected without the animals showing any clinically relevant signs. These carriers present a particular risk for breeding and blood transfusions. Non-haemotropic mycoplasmas can be found on the mucous membranes of the res- piratory and the urogenital tract, where they can escape from the immune response of Camelids the infected animal for a very long time. Conjunctivitis and rhinitis are clinically apparent, In its acute phase, an infection with Mycoplasma haemolamae can cause haemolytic disorders of the upper respiratory tract occur less frequently. Mycoplasmas can also be anaemia in affected animals. However, infections can also primarily progress silently primarily pathogenic. and lead to a chronic carrier state. The disease may fully break out in these animals in situations linked to stress and/or immunosuppression. Dog Mycoplasmas in dogs are often associated with diseases of the urogenital region and Pig with infertility. Clinically, an infection with canine mycoplasma can cause prostatitis and/ Porcine eperythrozoonosis is an infectious disease caused by Mycoplasma suis or orchitis in male dogs, and can, amongst others, lead to endometritis in female dogs. (formerly Eperythrozoon suis). The pathogens attach to the erythrocytes (adhesion, However, mycoplasmas may also play a role in canine respiratory diseases. invasion) and provoke damage to and lysis of the erythrocytes due to the formation 200 201 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Cat Pig In the cat common cold complex, not only viral components (FHV, FCV) play a role, but Mycoplasma hyopneumoniae is the primary causative agent of enzootic porcine also Mycoplasma felis. Clinically, an infection is usually manifested by conjunctivitis and pneumonia (EPP) in pigs. EPP is one of the most significant causes of respiratory infec- rhinitis. Mycoplasma gatae and Mycoplasma feliminutum are sometimes isolated from tious diseases in pigs. The disease is globally distributed. However, the pathogen only cats, nevertheless, their clinical relevance is questionable. As it is difficult to cultivate causes high economic losses in pig production when combined with poor environmen- mycoplasma, PCR detection is the method of choice. tal conditions and secondary bacterial and/or viral infections. Rat and mouse Poultry Mycoplasma pulmonis is the causative agent of “murine respiratory mycoplasmosis” Infections with Mycoplasma gallisepticum cause the so-called chronic respiratory in rats and mice, a slowly progressing infection of the respiratory tracts associated with disease (CRD) in chickens and infectious sinusitis in turkeys. Infection occurs both ho- the formation of thick mucus. Clinical signs of infected animals are sneezing, mucopu- rizontally through the air and direct contact as well as vertically through hatching eggs. rulent nasal discharge, stertorous breathing sounds and dyspnoea. The infection can The main symptoms include chronic inflammation of the upper respiratory tracts and spread to the middle ear and lead to otitis media and head tilt. air sacs, accompanied by disorders of the joints, tendon sheaths and the genital tract. In addition, especially in older female rats, Mycoplasma pulmonis can cause genital Central nervous disorders can also arise. In addition, laying performance and hatching infection which leads to infertility or a small litter size. In rare cases, metritis or pyometra rates decrease significantly. Mixed infections with viral pathogens, such as Newcastle are also seen. disease virus (NDV) or infectious bronchitis virus (IBV) are not unusual and can severely Latent infections without any clinical symptoms are common. aggravate the clinical picture (also as vaccination viruses). Transmission occurs through aerosols in close direct contact. Sexual or intrauterine transmission is also possible. In chickens and turkeys, Mycoplasma synoviae causes infectious synovitis and arthri- tis, which is clinically manifested by joint swellings and lameness. Inflammations of the Reptiles air sacs, myocardium and pericardium also occur. Especially after mixed infections, re- Several Mycoplasma spp. exist in tortoises. An infection with a virulent Mycoplasma spiratory symptoms can be seen as well. Stunted growth, reduced laying performance agassizii strain causes the so-called upper respiratory tract disease (URTD), a disease and greenish diarrhoea are also due to the infection. Besides game birds, geese, too, clinically characterised by serous, mucous and purulent nasal discharge as well as are susceptible to this pathogen. ocular discharge, conjunctivitis and eyelid oedema. Furthermore, it can cause lethargy, dehydration, anorexia and fatal cachexia. An essential trait of mycoplasma infections is Mycoplasma – Pathogen Detection the fact that they can persist in the organism without triggering any symptoms. Often, the disease only breaks out if there are other microorganisms and environmental fac- Material Dog: swab without medium (eye, pharynx, nose, genital tract), tors involved, combined with the genetic properties and immune reactions of the host. bronchoalveolar lavage, abortion material Mycoplasmas are also detected in turtles and other reptiles, but little is known about Cat: swab without medium (eye, pharynx, nose, genital tract), their clinical relevance. bronchoalveolar lavage, abortion material Mouse, rat: swab without medium (nose, pharynx), tissue (lung) Tortoise/turtle: swab without medium (conjunctiva, mouth), nasal Cattle flush In the first weeks of the life of a calf, Mycoplasma bovis can cause mostly enzootic Cattle: swab without medium (nose, pharynx), nasal flush, pneumonia and arthritis, and in cows it leads to severe mastitis. bronchoalveolar lavage, milk, synovia, sperm, tissue (lung) Affected calves typically suffer from otitis with hanging earlobes and head tilt. As mas- Pig: swab without medium (trachea, nose), bronchoalveolar titis pathogen, Mycoplasma bovis is highly infectious. Typically, the mammary gland lavage, tissue (lung) increases in size and hardens, and within a few weeks, the inflammation spreads to the Poultry: swab without medium (pharynx, cloaca), faeces, tissue neighbouring udder quarters. (lung), synovia The pathogen is also often detected in connection with chronic diseases of the respira- Method PCR/realtime PCR tory tract. Reservoirs of M. bovis are the respiratory tract of clinically healthy calves and Species Dog, cat, rat, mouse, tortoise/turtle, cattle, pig, poultry young cattle as well as the udder of cows with subclinical mastitis. Duration 1 – 3 days 202 203 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS Note This PCR detects at least M. arginii, M. gateae, M. spumans, M. Pasteurella multocida (toxin producing) – Pathogen Detection cynos, M. molare, M. canis, M. edwardii, M. bovigenitalum, M. maculosum, M. opalescens, M. feliminutum Material Swab without medium (nose, pharynx, conjunctiva), bronchoalveolar The detection of Mycoplasma bovis by PCR is also part of our Bovine lavage, tissue (lung) Respiratory Profile. Method Realtime PCR Species Rabbit, pig Duration 1 – 3 days Mycoplasma bovis – Antibodies Note The serogroups/types (A - F)/serotypes cannot be differentiated by Material S 1 ml PCR, as there is no correlation between the serotype and genotype Method ELISA characteristics. Species Cattle Duration 5 days 13.2.27 Rhodococcus equi Mycoplasma hyopneumoniae – Antibodies Material S 1 ml Rhodococcus equi is a facultative pathogenic organism in soil or horse faeces. It is Method ELISA the most frequent pathogen in heavy lung disease in foals aged 3 weeks to 6 months; Species Pig lethality is high. Entry and predilection site is the lung (abscessation!); hematogen Duration 2 – 3 days spreading in other organs is possible. Other entries may be the gastrointestinal tract (diarrhoeas and ulcers) or umbilical infections. Moreover, R.equi shows an affinity for Neoehrlichia mikurensis Ø see Candidatus Neoehrlichia mikurensis bones and joints. Detection by bacteriological culture allows to perform an antibiogram. (Chapter 13.2.10) Rhodococcus equi – Pathogen Detection Nocardia Ø see Chapter 14.4 Material (1) Swab with medium (nose, navel), bronchoalveolar lavage, Paenibacillus larvae Ø see Chapter 14.4 tracheal lavage (2) Swab without medium (nose, navel), bronchoalveolar lavage, tracheal lavage, faeces Method (1) Culture 13.2.26 Pasteurella multocida (2) Realtime PCR Species Horse Pasteurella multocida is a gram-negative bacillus. Pasteurella are commensals of the Duration (1) 3 days mucous membrane of the upper respiratory tract. Factors that reduce resistance, such (2) 1 – 3 days as overpopulation or a bad stable environment, provide a predisposition to infections with toxigenic strains. Co-infections with Bordetella bronchiseptica are common and lead to particularly severe symptoms. Pasteurella multocida, either as a monoinfection or together with Bordetella 13.2.28 Rickettsia bronchiseptica, leads to “snuffles” in rabbits. Normally, this disease is a stock problem and is often recurrent. Rickettsia are obligate intracellular coccoid, rod-shaped or pleomorphic gram-negative In pigs, Pasteurella multocida toxin is the aetiological agent that causes progressive bacteria that parasitise in reticuloendothelial cells or erythrocytes. They are usually atrophic rhinitis, with especially the toxigenic pasteurella types A and D being transmitted by arthropods. involved. The cytotoxic toxin (PMT) inhibits the osteoblasts. With the activity of the Rickettsia are divided into the categories “spotted fever group”, thyphus group and osteoclasts being maintained, it leads to atrophy of the nasal conchae and deformation “others”, which includes Coxiella burnetii. of the nasal septum. The importance of the toxin in pneumonia in cattle and pigs has In the USA, Rickettsia rickettsii, the causative agent of “Rocky Mountain spotted not yet been clarified. fever”, and in the Mediterranean area, Rickettsia conorii, the causative agent of “Mediterranean spotted fever”, are of central importance in animal infections. Infected 204 205 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS dogs may remain asymptomatic or show symptoms ranging from lymphadenopathies, Germany: In cattle, it is an epizootic disease that is notifiable upon suspicion. In fever, hyperesthesia, peripheral oedema up to lameness. In laboratory diagnostics, other species, it is notifiable upon diagnosis. neutrophilic lymphocytosis with variable left shift, thrombocytopenia, hypoproteinaemia and hypergammaglobulinaemia are found, among others. Salmonella – Pathogen Detection Material (1) Faeces (swab with medium: intestine, cloaca) Rickettsia conorii – Antibodies (2) Faeces; birds: faeces, swab without medium (cloaca), eggs, Material S, EP, HP 0.5 ml tissue Method IFAT Method (1) Culture including enrichment, MALDI-TOF Species Dog (2) Realtime PCR Duration 1 day Species All Note Not endemic in Northern Europe, we find R. conorii in the Duration (1) 2 – 3 days Mediterranean, Africa, South West Asia, India. Serological studies (2) 1 – 3 days suggest a high prevalence in asymptomatic dogs. Note A faecal sample of 2 cm of diameter or larger is required. In exceptional cases, a swab with transport medium can be used Rickettsia rickettsii – Antibodies too. Material S, EP, HP 0.5 ml Method IFAT Salmonella – Antibodies Species Dog Material S, EP 1 ml Duration 1 day Method Agglutination (Gruber Widal reaction) Note Rickettsia ricketsii infection causes Rocky mountain spotted fever. It Species Dog, cat, birds, horse is found in North and South America. Duration 2 days Note Direct detection by culturing Salmonella from faecal samples is to be preferred greatly over any indirect detection. 13.2.29 Salmonella Salmonella abortusequi – Antibodies* Salmonella/Shigella Ø see also Chapter 16.1.2 Material S 1 ml Method Slow agglutination Salmonella belong to the family Enterobacteriaceae and are found in the intestines of Species Horse animals and humans. In most cases, infection occurs faecal-orally or by feeding raw Duration 5 days meat. Salmonella infections affect almost all animal species. Compared to herbivorous pets, Note Export-relevant test. dogs and cats are more resistant to salmonella infections. Under favourable conditions, In the host-adapted serovar Abortusequi, pathogen transmission salmonellosis causes diarrhoea with vomiting and fever; in young animals, the disease occurs orally; rarely through mating. With regard to miscarriages, can also become septicaemic. In reptiles and amphibians, salmonella can be part of this pathogen does not currently play a role in Germany anymore. the normal intestinal flora. In these animals, clinically relevant salmonellosis is associa- ted with immune deficiency. Shigella Ø see Salmonella/Shigella in Chapter 16.1.2 According to the Robert Koch Institute (RKI), about 10% of all human salmonella infec- tions, which cause diarrhoea, are related to direct contact with excreting dogs, cats and particularly reptiles. For some time now, ESBL producers have also been detected among salmonella, especially in livestock. Because of the ESBL problem, creating an antibiogram is essential. 206 207 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS 13.2.30 Staphylococcus the same time offering a comparably higher sensitivity and specificity. This way, the identification of clinically healthy carriers, which play a major role in pathogen Staphylococci are gram-positive and extremely resistant bacteria. They normally reside epidemiology, is also more reliable. on the skin and the mucous membranes, where they are part of the physiological germ As PCR does not differentiate between dead or living organisms, a positive pathogen flora. detection should always be formulated as a suspected diagnosis and be confirmed by Inflammations caused by staphylococci are usually locally limited. Only in cases of decre- culture examination. ased resistance, septicaemia and pyaemia can occur. In ruminants, staphylococci are of Clinically, an infection with Streptococcus equi subsp. equi cannot always be major importance as causative agents of mastitis. distinguished from an infection with Streptococcus equi subsp. zooepidemicus. Nowadays, special attention should be paid to whether methicillin-resistant strains of Streptococcus equi subsp. zooepidemicus can be found in all domestic animals and Staphylococcus aureus (MRSA) or, in the small animal practice, of Staphylococcus in humans. In horses, it is a facultative pathogenic commensal; infections can cause, pseudintermedius (MRSP) are present. If there is a corresponding resistance pattern, we amongst others, respiratory disorders and purulent bronchopneumonia. As with detect the mecA gene by means of PCR. In case of repeated wound healing problems in strangles, especially foals and young horses are affected. patients visiting the practice, which are caused by MRSA or MRSP, it should be consi- dered testing the practice‘s personnel, too, whether they carry this type of germ on their Streptococcus equi – Pathogen Detection nasal mucosa. Material (1) Swab with medium (nose, abscess, lymph node), BAL, TBS, Detection can be done through culture examination of clinical samples, e.g. swabs of guttural pouch lavage sample pustules, mucosal swabs and other body secretions and excretions. (2) Swab without medium (nose), lavage sample (BAL, TBS, guttural pouch), tissue (lymph node) Staphylococcus – Pathogen Detection Method (1) Culture Material Swab with medium, milk (ruminants) (2) Realtime PCR Method Culture including enrichment Species Horse Species All Duration (1) 2 – 3 days Duration 3 days (2) 1 – 3 days Note Further differentiation can be made if MRSA is suspected. Note In culture, both subspecies (Streptococcus equi equi and Strepto- coccus equi zooepidemicus) are determined and differentiated. If detection is to be done by means of PCR, it can be chosen Staphylococcus – Antibodies between the single detection of Streptococcus equi equi or the Material S 0.5 ml detection of both subspecies mentioned above. Method Agglutination Species Dog, cat Streptococcus equi equi – Antibodies Duration 1 day Material S 0.5 ml Note To recognise sensitised animals in cases of pyoderma. Method ELISA Species Horse Duration 1 day 13.2.31 Streptococcus equi Note Generally, this test determines both Streptococcus equi equi and The globally spread and highly infectious equine disease strangles is caused by an Streptococcus equi zooepidemicus. However, the detected surface infection with Streptococcus equi subsp. equi and is characterised by purulent antigen SeM is considered a virulence factor which mainly occurs in lymphadenitis and pharyngitis. It is a typical disease in young animals which induces Streptococcus equi equi. long-lasting immunity; thus, older animals rarely contract the disease. Over the past years, however, an increasing number of affected adult horses has been described, with the disease showing a rather atypical progression (mainly fever, respiratory disorders). Compared to culture, PCR has the advantage of delivering faster results while at 208 209 2019/20 Infectious Diseases / Bacteria 2019/20 Infectious Diseases / Bacteria LABORATORY FOR CLINICAL DIAGNOSTICS 13.2.32 Taylorella equigenitalis Treponema paraluiscuniculi – Pathogen Detection Material Tissue (mainly skin lesions/scurfs; if need be regional lymph nodes), Contagious equine metritis (CEM) is caused by the gram-negative bacillus Taylorella swab without medium (vagina, prepuce) equigenitalis. Transmission particularly occurs during mating; stallions latently carry the Method Realtime PCR pathogen on the mucous membrane of the penis, especially in the Fossa urethralis and in Species Rabbit, hare the smegma of the prepuce. Transmission from infected mares to stallions is also possible. Duration 1 – 3 days In mares, an infection leads to endometritis/cervicitis with mucopurulent vaginal discharge and to reduced fertility. Stallions show no clinical signs of the disease. Within the EU, the detection of Taylorella equigenitalis by PCR is also considered an Treponema paraluiscuniculi – Antibodies* appropriate test method in addition to the bacteriological examination. We recommend Material S 0.5 ml it as a quick way to get a preliminary diagnosis; to receive the official export licence, a Method Treponema pallidum haemagglutination test bacteriological examination is still required. In Germany, there is an obligation to inform Species Rabbit the authorities, if Taylorella equigenitalis is detected. Duration 1 week Taylorella equigenitalis/CEM – Pathogen Detection Material (1) Swab with medium (Amies with activated charcoal, ≤ 48 h) 13.2.34 Yersinia (2) Swab with medium, sperm Stallion: penile sheath, urethra, fossa glandis Yersinia belong to the family Enterobacteriaceae. Yersinia (Y.) pseudotuberculosis Mare: fossa clitoridis, sinus clitoridis is the causative agent of pseudotuberculosis/rodentiosis, an infectious disease Method (1) Culture, MALDI-TOF all mammalian and bird species can contract. For instance, rodents and cats are (2) Realtime PCR predisposed. Abscesses can occur in various organs. The pathogen has a high Species Horse tenacity. In the soil, the pathogen remains infectious for months. Duration (1) Culture 1 week Y. enterocolitica causes enterocolitis in humans and animals. Immunopathological Export to Canada: 2 weeks, reactions can lead to arthritis, arthrosis and skin diseases. Animals, especially pigs and Export to Norway: 3 weeks poultry, often act as pathogen reservoir. Dogs rarely become ill, if at all, puppies are (2) PCR 1 – 3 days mainly affected. The infection manifests itself as enteritis, resulting in mucous to bloody diarrhoea. Note Detection by PCR is offered as an individual service for one sample or as a profile for 2 samples (mare) resp. 3 samples (stallion). Yersinia – Pathogen Detection Material (1) Faeces (swab with medium) 13.2.33 Treponema paraluiscuniculi (2) Faeces, swab without medium (rectum) Method (1) Culture including enrichment Rabbit syphilis (Spirochaetosis cuniculi) is caused by the highly contagious bacterium (2) Realtime PCR (only Y. enterocolitica) Treponema paraluiscuniculi. Only rabbits and hares are susceptible, human infections Species All are not possible. Transmission occurs directly, usually during mating. However, the Duration (1) Up to 3 weeks animals can also become infected through other mucosal contacts as well as bedding (2) 1 – 3 days and feed. Rabbit syphilis typically occurs as a chronic disease, but latent infections are Note A faecal sample of 2 cm of diameter or larger is required. possible, too. The incubation period is weeks to months. First clinical symptoms can be In exceptional cases, a swab with transport medium can be used too. seen as oedematous swellings and formation of nodules on the external genital organs. Detection by culture is offered as a combined service together with In the further course of the disease, these nodules erode into ulcers and become the detection of Campylobacter. purulent and encrusted. By licking the affected anogenital region, other skin areas, such as lips, eyelids or the edge of the ears often become infected as well. 210 211 2019/20 Infectious Diseases / Bacteria, Fungi 2019/20 Infectious Diseases / Fungi LABORATORY FOR CLINICAL DIAGNOSTICS Yersinia – Antibodies Duration 1 day Material S 0.5 ml Note Cultural detection of Aspergillus is often very difficult due to the site of Method Agglutination infection. Antibody detection may be used to support the diagnosis. Species Dog, cat Duration 1 – 3 days 13.3.2 Batrachochytrium Note Detection of antibodies against Yersinia enterocolitica and Yersinia pseudotuberculosis. Batrachochytrium are fungi and being held responsible for large losses in amphibians. Batrachochytrium dendrobatidis 13.3 Fungi The chytrid fungus Batrachochytrium (B.) dendrobatidis was first identified in Australia in 1998 and named in 1999. This fungus is thought to be responsible for the population 13.3.1 Aspergillus decline and the global extinction of >200 amphibian species. Infections with B. dendrobatidis are in many cases associated with very high mortality Aspergillus is a mould with approximately 200 species found worldwide. In the rates (in lab up to 100%), but the fungus is not necessarily lethal. Other factors such as environment, Aspergillus is particularly found in compost heaps and in the ground. stress or co-infections with other pathogens also appear to play a role. Aspergillosis is often caused by the species Aspergillus fumigatus and preferably B. dendrobatidis multiplies in keratinized tissue and therefore affects primarily the outer affects the skin, nose, paranasal sinuses and the lung. Aspergillosis frequently occurs skin of adult animals (stratum corneum to the stratum granulosum). in birds, especially if the animal is predisposed by improper keeping, the administration In larvae, the horn strips on the mouth are affected. During metamorphosis the infections of antibiotics or stress. This very often results in severe respiratory disorders. Other can lead to dramatic high mortality rates. The clinical symptoms are often nonspecific organs (e.g. CNS) can also be affected. and may, in addition to the skin (often appear macroscopically unchanged or „blunt“ or depigmented; hyperkeratosis and massive skinning episodes, mixed infections with severe erosions of the skin) or behavioural changes (atypical behaviour, such as Aspergillus-Galactomannan – Antigen Detection* prolonged stay in the water, ataxia and CNS problems). Spontaneous deaths without Material S 0.5 ml previous overt clinical disease are also observed. Method ELISA Species Birds Batrachochytrium dendrobatidis – Pathogen Detection Duration 1 week Material Swab without medium: skin swabs of the ventral body surface (adult Note Galactomannan is a polysaccharide found in the cell wall of Asper- animals) or rather of the keratinized skin at the mouth (tadpoles), gillus spp. In animals affected by aspergillosis, it can be detected in tissue (skin frazzles of infected animals) the blood. Method PCR For birds that only develop low antibody titres against Aspergillus Species Amphibians (often in parrots), the detection of galactomannan in the blood can Duration 1 – 3 days help to diagnose aspergillosis. Animals that display high antibody titres against Aspergillus normally do not have detectable galactomannan in the blood, so that both Batrachochytrium salamandrivorans tests in combination can complement each other when making a Batrachochytrium salamandrivorans is a recently described highly-contagious and diagnosis. deadly chytrid fungus that has massively infested and killed fire salamanders especially in North-west Europe. Infected animals show anorexia, apathy and ataxia as well as skin lesions with super- Aspergillus sp. – Antibodies ficial erosions and deep ulcerations all over the body. Suitable test materials are skin Material S 0.5 ml biopsies and swabs. Method Agglutination Species Dog, cat, birds, cattle, others on request 212 213 2019/20 Infectious Diseases / Fungi 2019/20 Infectious Diseases / Fungi, Parasites LABORATORY FOR CLINICAL DIAGNOSTICS Batrachochytrium salamandrivorans – Pathogen Detection 13.3.4 Macrorhabdus ornithogaster Material Swab without medium (skin), tissue (skin) Macrorhabdus ornithogaster is a yeast which is found in many different avian species, Method PCR especially in budgerigars. Macrorhabdiosis is also called megabacteriosis or going Species Amphibians (mainly salamander) light syndrome. Infected animals may develop maldigestion and lose weight even Duration 1 – 3 days though there is no change in appetite. Undigested seeds can be excreted with the faeces, sometimes there is also blood in the faeces. Choking and vomiting as well as a general weakness can also be found in macrorhabdiosis. Inapparent carriers are fre- 13.3.3 Dermatophytes quent. Transmission presumably occurs through billing and feeding as well as through contaminated feed and water. Dermatophytes are filamentous fungi that can cause skin lesions in humans and animals. The disease is called dermatophytosis. The fungi use keratin as a carbon source and colonise keratinized tissue (hair, skin, nails). Macrorhabdus ornithogaster – Pathogen Detection Dermatophytes are highly contagious. Infection is direct or indirect. Enabling factors Material Faeces, proventriculus, smear on slide are, for example, immunosuppression, reduced immune response (e.g. high age) or Method Stain, microscopic pre-damage of the skin (e.g. by ectoparasites). Spores as propagating form can also Species Birds remain infectious in the environment for years. Duration 1 day The clinical symptoms are diverse and dependent on the virulence of the fungal strain, the infection period and the immune status of the host. Typical patchy alopecia may appear on the face, ears and front legs. An itch may be missing or from mild to severe. 13.3.5 Ophidiomyces In case of skin diseases, a dermatophytosis must always be considered in the differential diagnosis. Ophidiomyces (formerly Chrysosporium) ophiodiicola is a causative agent of snake fungal disease. Infections with O. ophiodiicola are associated with skin lesions, pustu- Dermatophytes – Pathogen Detection les, nodules and swelling of the skin. Lesions primarily occur on the head, but can also spread to the whole body. Material Hair with hair roots, deep skin scraping, scales, crusts, claws Method (1) Culture (2) PCR Ophidiomyces ophiodiicola – Pathogen Detection Species Dog, cat, rabbit, guinea pig, horse (and other species) Material Swab without medium (skin), tissue (skin) Duration (1) 3 days to 4 weeks Method Realtime PCR (2) 2 – 4 days Species Snake Note • Culture also detects Malassezia; a false negative result is possible Duration 1 – 3 days with previous treatment. • PCR is validated for the detection of the following dermatophyte species: Microsporum canis, Microsporum gypseum, Microspo- 13.4 Parasites rum persicolor, Trichophyton mentagrophytes, Trichophyton equi- num. Further types of skin fungus might also be detected by the PCR. Differentiation of the most common dermatophyte species 13.4.1 Angiostrongylus vasorum can be conducted on request. Angiostrongylus vasorum is a globally distributed nematode that parasitises the • PCR is not suited for therapy monitoring (dead dermatophytes are pulmonary arteries and, less frequently, the right heart of dogs and wild canines. detected as well). Infections with A. vasorum occur more often in Germany than normally expected (a prevalence of 7.4% according to Barutzki and Schaper, 2009). Thus, an infection with this lungworm should always be considered in differential diagnosis if respiratory and/or cardiovascular symptoms are present. 214 215 2019/20 Infectious Diseases / Parasites 2019/20 Infectious Diseases / Parasites LABORATORY FOR CLINICAL DIAGNOSTICS Dogs as definitive hosts get infected by ingesting L3 larvae when eating infected snails of the animals over months, anaemia and intermittent periods of fever. or slugs (intermediate hosts). L3 invade the lymphatic and blood system through the Without treatment, dogs can also develop a subclinical form with the blood count being wall of the small intestine of the dog and enter the pulmonary arteries. Six to eight normal again. Many imported dogs from the south are subclinically infected and thus weeks p.i., the females begin to lay eggs. Via the blood, the eggs reach the fine pose a risk of infection for other dogs. In addition, the infection can be reactivated in pulmonary capillaries where they develop into L1 larvae and enter the pulmonary these dogs by various factors. Cattle and horses can also remain carriers of Babesia for alveoli. From here, they are carried up by the ciliated epithelium or are coughed up, many years. swallowed again and finally excreted with the faeces. L1 are taken up with the faeces by intermediate hosts and the infectious L3 then develop within them. Dog Babesia canis Especially young dogs between the age of one and two years are affected by canine B. canis is transmitted by Dermacentor reticulatus and is more virulent than B. vogeli. A angiostrongylosis. Besides clinically inapparent infections, the course of the disease distinction is made between the French and the Hungarian strain. may be mild to life-threatening. Clinical symptoms are highly variable, however, French strain: Distribution: north and east Mediterranean area, locally in Holland the main symptoms include cardiopulmonary signs such as dyspnoea and cough. (The Hague, Arnhem), focuses in western Germany (Saarland, Rhineland-Palatinate, The second most typical symptoms are coagulopathy with epistaxis, haemoptysis, Baden-Wuerttemberg). What is often noticed about the French strain is its low antibody haematoma and anaemia. Subsequently, DIC, circulatory insufficiency and death can production. occur. Vomiting or neurological symptoms like muscle tremor, ataxia, dizziness and Hungarian strain: Distribution: Hungary, Ukraine, Romania, eastern Germany. epileptiform seizures are also possible. What is often noticed about the Hungarian strain is its high antibody production. In 80% of the animals, new infections with the Hungarian strain lead to death if untreated. In Angiostrongylus vasorum – Pathogen Detection this strain, it is particularly important to achieve pathogen elimination with the treatment in order to counteract further spread of this strain. Material (1) Faeces: 3 days collective sample (2) EB, BAL, (faeces: 3 days collective sample), tissue (lung, brain) Babesia vogeli Method (1) Baermann-Wetzel method Distribution: North Africa, the whole Mediterranean area, Southern Europe, France. In (2) Realtime PCR Germany, 2 isolates from Dermacentor ticks were discovered in the Berlin metropolitan Species Dog area and the Elbe valley. Duration (1) 2 days B. vogeli is transmitted by Rhipicephalus sanguineus and often only leads to low anti- (2) 1 – 3 days body titres. Note If blood should be examined, a combination of PCR and ELISA is recommended as it increases sensitivity. Babesia gibsoni Distribution: Asia, USA, Europe (imported) Angiostrongylus vasorum – Antigen Detection Distribution in Europe is considered questionable. The cases of Babesia gibsoni de- scribed for Portugal and Spain were corrected later, partly into the pathogen Theileria Material S 0.5 ml annae. Method EIA Species Dog Duration 1 day Cat Babesia canis Distribution: Thailand, Brazil, France, Poland, Germany 13.4.2 Babesia Babesia felis Distribution: in parts of Africa Babesiosis in mammals has become one of the most important parasitic diseases. The pathogens, which belong to the order Piroplasmida, are transmitted by ticks. Babesia cati In peracute or acute infections, unspecific clinical signs such as fever, apathy and loss Distribution: India of appetite appear between the 5th and 28th day p.i. Anaemia, icterus and massive haemoglobinuria occur. A chronic infection is characterised by fatigue and emaciation 216 217 2019/20 Infectious Diseases / Parasites 2019/20 Infectious Diseases / Parasites LABORATORY FOR CLINICAL DIAGNOSTICS Horse Babesia – Antibodies Babesia caballi and Theileria equi (formerly Babesia equi) Distribution: From the tropics and subtropics to the temperate zones. Equine Material S, EP, HP 0.5 ml; B. gibsoni: S 0.5 ml babesiosis (piroplasmosis) is also expected to occur in Germany. Method (1) Cat, horse: IFAT (2) Dog: ELISA (3) Horse: c-ELISA (export to USA, most sensitive test) Cattle (4) Horse: CFT* (export to most countries) Babesia divergens Species Dog, cat, horse Distribution: in Europe from Finland down to the Mediterranean Duration (1 and 2) 1 day (3) 2 – 3 days Babesia major (4) 5 days Distribution: Central Europe in small endemic areas Note Seroconversion from the 2nd week p. i., maximum titre after 4 Babesia bigemina weeks. Distribution: tropics and subtropics; in Europe: the Balkans, coastal areas in the False negative results can occur in early phase of infection and in Mediterranean, Portugal rare cases esp. young dogs (< 6 months). Upon request, the detection in dogs can also be carried out by IFAT. Babesia – Pathogen Detection Material (1) EB 1 ml + blood smear 13.4.3 Coccidia (2) EB 0.2 ml, tick Method (1) Microscopic evaluation Tortoise intranuclear coccidiosis (TINC) is a severe disease in tortoises with high (2) Realtime PCR morbidity and mortality rates. TINC has already been detected in different tortoises and Species Dog, cat, horse, cattle box turtles in North America and Europe. Clinical symptoms include lethargy, signifi- Duration (1) 1 day cant weight loss, erosive rhinitis, dyspnoea and occasionally skin lesions. Infections (2) 1 – 3 days are generally systemic. These coccidia are most frequently detected in the intestine, Note (1) Microscopic detection is possible from the 5th day post pancreas, liver and kidney. However, they can also be found in the Eustachian tube, in infectionem. It is preferable to collect capillary blood (edge of the macrophages of the spleen, in the middle ear, lungs and stomach. In live animals with ear) and spread it onto a glass slide. The detection from capillary rhinitis, they can also be detected in nasal lavage samples. blood is much more sensitive! (2) PCR detection is far more sensitive than the detection from a Intranuclear Coccidiosis (TINC) – Pathogen Detection blood smear. In case of a chronic infection, it can be assumed that pathogens have spread to many sites. However, the Material Swab without medium (nose, if need be cloaca), nasal flush, tissue concentration of pathogen DNA in the blood may be very low (nose (mucosa), intestine, pancreas, kidney, liver) and thus lead to a negative result in the PCR. While a positive Method Realtime PCR PCR is proof of an infection, a negative PCR never rules out an Species Tortoise, turtle infection. Duration 1 – 3 days PCR horse: In case of a positive result, a differentiation between Theileria equi/Babesia caballi can subsequently be made on request. 218 219 2019/20 Infectious Diseases / Parasites 2019/20 Infectious Diseases / Parasites LABORATORY FOR CLINICAL DIAGNOSTICS 13.4.4 Cryptosporidia Cryptosporidia infection, as the pathogen can be excreted intermittently. Symptomatic therapy and hygiene management are the best options in the fight against Cryptosporidia are very small, unicellular parasites of the gastrointestinal tract. They are Cryptosporidium. classified as coccidia. Different species are described with very similar morphology. Some of these are host-specific; others (e.g. Cryptosporidium parvum) can infect Cryptosporidia – Pathogen Detection various animal species and humans (zoonosis). Material Faeces; snake: faeces, regurgitated material, gastric lavage, biopsy Infections occur after intake of sporulated oocysts. The infectious dose is very low (stomach) (approx. 100 oocysts). Subsequently the liberated sporozoites infect the intestinal Method (1) Antigen detection: EIA, IFAT (reptiles) epithelial cells, followed by a development cycle over trophozoites, meronts, (2) PCR merozoites, gamonts, zygotes and in the end again oocysts are formed. The oocysts (3) modified Ziehl-Neelsen staining excreted in the faeces show a high tenacity, are resistant to many disinfectants and Species Dog, cat, small mammals, reptiles, cattle, other species can remain infectious for months. Therefore, e.g. contaminated pens or terrariums are Duration (1) IFAT: 1 day, EIA: 2 days frequent sources of infection. (2) 2 – 4 days In cattle, Cryptosporidia is a very common endoparasite. A large proportion of calves (3) 1 day go through an infection with C. parvum. Clinically apparent courses with enteritis and diarrhoea occur especially in calves up to 3 weeks of life, often related to co-infections. Note If the PCR yields positive results in reptiles, it is possible to perform a Not infrequently, lambs, piglets and foals are affected. differentiation of the Cryptosporidium species to distinguish between A much lower prevalence is seen in dogs and cats, with usually asymptomatic harmless species of Cryptosporidium, which are ingested with the infections. However, oocysts are excreted in the faeces here too for about 2 weeks. prey, and pathogenic species. Manifest infections can be seen in puppies or immunosuppressed animals (e.g. FeLV, FIV, distemper, neoplasia etc.). In reptiles, Cryptosporidia is a serious pathogen that can cause severe losses, 13.4.5 Demodex especially in snake and lizard stocks. C. serpentis is an important parasite in snakes and infects the gastric mucosa. Due to the chronic inflammation a subsequent swelling Demodex mites are strictly host-specific ectoparasites of numerous mammals and of and hardening of the connective tissue in the gastric area can occur. Typical symptom humans. So far, there have been three species each described in dogs and cats (dogs: is regurgitation of food days after digestion. C. saurophilum (also called C. varanii) particularly Demodex canis, rarely D. injai and D. cornei; cats: especially D. cati, but however destroys the lining of the intestinal walls of affected lizards and snakes. also D. gatoi and only recently D. felis, too). Clinically malabsorption with excretion of undigested food, profound weight and fluid The entire development of Demodex mites takes place in the hair follicles, the loss is observed. Both pathogens are not pathogenic to humans. Quite often, C. muris sebaceous and apocrine glands of the host. They cannot survive very long in the and C. parvum are found as passers in reptile faeces (origin: infected food animals). environment. Transmission mainly occurs postpartum while nursing. Demodex mites Therefore, a differentiation is absolutely necessary by a positive Cryptosporidium result. belong to the physiological skin fauna, but are potentially pathogenic. In dogs, there is often a low number of mites present without any clinical symptoms (prevalence up to Laboratory diagnostically several methods are available for detection. Already during 85%), but demodicosis is rare. Nevertheless, it is one of the most frequent dermatoses the microscopic examination after specific enrichment (MIFC) oocysts can be found. in dogs (especially young dogs), in cats, however, it is very rare. Like all parasitological faecal examinations, the sensitivity here is relatively limited by In dogs, lesions generally start in the face or on the forelegs and spread from there. around 60%. The localised form affects a few well-defined skin areas and most notably occurs in In cattle, ELISA testing (detects C. parvum) is recommended. The immunofluorescence young dogs. The skin areas are often hairless and may also be scaly. Comedones are test includes a wider range of Cryptosporidium species and is therefore suitable for typical as well. In general, itching only occurs in case of secondary bacterial infections. dogs, cats, but also small rodents (guinea pig: C. wrairi). If more than four lesions are present, an entire body region or at least two paws are Reptile faecal samples are additionally stained (modified Ziehl-Neelsen) to increase affected and if it continuously worsens without treatment, it is referred to as generalised the detection rate upon microscopic examination. In reptiles, one cannot distinguish demodicosis. There are usually secondary bacterial infections present and alopecia between pathogen strings or passers by positive IFAT results. Here, PCR testing is appears with follicular papules up to furunculosis, focal ulcerations and fistula tracts. recommended with subsequent differentiation. Most of the time, there is no itching, but sometimes intense pain. Fever, anorexia, It should be noted that a single negative result does not completely rule out a lethargy, lymphadenopathy and sepsis may occur and might be fatal if not treated. Special forms are podo- and otodemodicosis. 220 221 2019/20 Infectious Diseases / Parasites 2019/20 Infectious Diseases / Parasites LABORATORY FOR CLINICAL DIAGNOSTICS Favourable factors for mass reproduction of mites include, e.g., endoparasitosis, Encephalitozoon cuniculi – Pathogen Detection malnutrition, cortisone treatment, neoplasia, hypothyroidism or hyperadrenocorticism. There is a genetic predisposition in young dogs (juvenile generalised demodicosis). Material Urine, CSF 0.2 ml, tissue (e.g. kidney, brain or eye/lens) These dogs should be excluded from breeding. Method PCR In cats, demodicosis particularly occurs if systemic diseases, such as diabetes Species Rabbit, guinea pig and others mellitus, FIV, FeLV or neoplasia, are present, and most notably causes alopecia and Duration 1 – 3 days crusts on the head and neck. Itching is also possible. Encephalitozoon cuniculi – Antibodies Demodex – Pathogen Detection Material S, EP, HP 0.5 ml Material Deep skin scraping Method IFAT Method (1) Microscopic evaluation Species Dog, rabbit, guinea pig (2) Realtime PCR (semi-quantitative) Duration 1 day Species Dog, cat Note Positive titres can be expected from day 14 post infection on. Duration (1) 1 day Subclinical infections are possible. (2) 1 – 3 days Note • Since demodex mites belong to the normal skin fauna and only Fasciola hepatica Ø see Chapter 15.2 an excessive increase leads to demodicosis, a positive PCR result should always be interpreted in connection with clinical and epidemiological data. A negative PCR result cannot completely 13.4.7 Filaria rule out an infection. • If reduced immunocompetence or immunodeficiency is In Europe alone, five different filarial species are known to cause filariasis in dogs: suspected, examination of the lymphocyte subpopulation by flow Dirofilaria immitis, Dirofilaria repens as well as Dipetalonema reconditum, Dipetalonema cytometry may be helpful. (Acanthocheilonema) dracunculoides and Cercopithifilaria grassi. Dirofilaria immitis causes „cardiovascular dirofilariasis“ (heartworm disease), Dirofilaria repens causes Echinococcus Ø see Chapter 15.2 „cutaneous dirofilariasis“. Both types of dirofilariasis are zoonoses and are transmitted by mosquitoes, including the common house mosquito (Culex pipiens) which is very common in Germany. According to literature, the other three filarial species are 13.4.6 Encephalitozoon cuniculi considered apathogenic, a fact which will not be tenable in future, at least concerning Dip. reconditum and possibly Dip. dracunculoides, too. The pathogen Encephalitozoon cuniculi causes encephalitozoonosis (also called torti- collis, wry neck, head tilt) in rabbits. Approximately 80% of healthy rabbits carry the pa- Dirofilaria immitis – Antigen Detection thogen without showing any clinical signs. Mature infectious spores are mainly excreted in the urine, so that transmission takes place orally and nasally by eating infected food Material S, EP, HP 0.5 ml or sniffing at food and litter. However, infected pregnant female hares can also transmit Method Immunoassay the pathogen to their young in the womb. Faecal shedding of pathogens was detected Species Dog, cat, ferret but seems to be of little importance. Duration 1 day The pathogen has also been found in many other animal species such as dogs, foxes, Note The serological examination is the most sensitive detection method rodents and some bird species and even in humans. Especially in immunocompro- for Dirofilaria immitis and detects the surface proteins of female, mised persons, infection can be relevant. parturient filariae (macrofilarial worms), which are parasites in the Apart from head tilt, the clinical picture in rabbits is mainly characterised by ataxia, heart or in larger vessels. The earliest time for a positive result is nystagmus, seizures or cramps. As the disease can also take a milder course, it is half a year p.i., but it can be delayed up to nine months if infected recommended to test for E. cuniculi in case of any neurological sign. dogs receive heartworm prevention. Examination of puppies under the age of six months is therefore not appropriate. If in doubt, it is 222 223 2019/20 Infectious Diseases / Parasites 2019/20 Infectious Diseases / Parasites LABORATORY FOR CLINICAL DIAGNOSTICS recommended to test for microfilariae or to take the test at a later Giardia – Pathogen Detection point in time. Therapy monitoring should be done at the earliest 4 – 5 months after completing therapy. For the detection of microfilariae Material Faeces see below. Method (1) Microscopic evaluation after enrichment (2) EIA (antigen detection) (3) Realtime PCR Microfilaria – Pathogen Detection Species Dog, cat, small mammals, Material EB 0.5 ml reptiles, large animals Method Microscopic evaluation, Knott test, filtration test, realtime PCR (semi- Duration (1 and 2) 1 day quantitative) (3) 1 – 3 days Species Dog, cat Note • Giardia infections lead to a Ferret (PCR) decrease in vitamin B 12. Duration 1 – 3 days • If treatment of Giardia fails in Note Accumulation of the microfilariae of Dirofilaria immitis in the cats, Tritrichomonas foetus peripheral blood takes place in the evenings (adaptation to the should also be considered. piercing behaviour of vector mosquitoes). This behaviour has not yet • In case of a positive PCR result, been documented for other filarial species, but it is reasonable to testing for the presence of human-pathogenic assemblages A take the blood sample after 4 pm. and B can subsequently be conducted. Before departing to South Africa, it is mandatory to do a filtration test. In case of a positive PCR result, differentiation of the filarial species can be done on request. 13.4.9 Hepatozoon Hepatozoon canis belongs to the protozoa and goes through a typical coccidial life 13.4.8 Giardia cycle with the dog as intermediate host. Asexual reproduction, schizogony, takes place in several generations in the endothelial cells of the spleen, liver and bone marrow. The Giardia is a flagellate that can be found in the intestine of mammals, birds, reptiles, merozoites formed here penetrate the leukocytes and differentiate into gamonts. amphibians and humans. There are some well-differentiated species, such as G. The definitive host, the tick, ingests the gamonts during the blood meal. Gamogony and intestinalis (lamblia, duodenalis). Giardia is ingested orally (food, water) or through sporogony take place in the tick and oocysts with 16 infectious sporozoites each are smear infection as cysts, excystates in the intestine and attaches as trophozoites to formed. the intestinal wall where it replicates. Damage to and detachment of the intestinal Infection with H. canis occurs by biting or swallowing an infected tick, primarily the brown epithelium cause chronic intermittent catarrhal to mucous-bloody diarrhoea. The cysts dog tick (R. sanguineus), which is found in warm countries (mainly Southern Europe, that are excreted with the faeces remain infectious for many months in cold water and a South America, Africa and Asia). By now, the pathogen has also become endemic in humid environment. several regions of Germany. Horizontal intrauterine transmission is possible as well. Except for Giardia in birds and amphibians, Giardia is partially of zoonotic nature. Seven Acute infections are characterised by fever, lymphadenitis, anorexia, apathy, myositis variations have been identified through genetic characterisation of which variations and epileptiform seizures (bleeding in meninges). Massive lesions up to necrosis (assemblages) A and B mainly occur in humans, variations C and D are primarily occur in the affected organs (spleen, liver, lung, brain). Chronic infections cause detected in dogs and variation F can mostly be found in cats. intermittent fever, lymphadenopathy, anaemia, diarrhoea and vomiting. Hyperaesthesia Across species, however, isolates of different subtypes of A as well as those of B can and muscular pain with stiffening of the neck muscles and the trunk muscles occur. be detected in different animal species, so that a transmission from humans to animals Periosteal bone proliferation can occur. In case of low parasitaemia, the infection may be and from animals to humans cannot be excluded. In dogs and cats, Giardia is the clinically inapparent or may only have mild clinical signs. predominant type of intestinal parasites. In our own examinations, Giardia infections were detected in 15% of cats; 3.5% of these animals contained the human-pathogenic Hepatozoon – Pathogen Detection assemblage A. Material EB 0.2 ml, tissue (liver), tick Method (1) Microscopic evaluation (buffy coat) (2) Realtime PCR (Hepatozoon canis/felis) 224 225 2019/20 Infectious Diseases / Parasites 2019/20 Infectious Diseases / Parasites LABORATORY FOR CLINICAL DIAGNOSTICS Species Dog, cat Note Positive antibody titres appear earliest 2 – 3 weeks post infection. Duration 1 – 3 days In asymptomatic dogs, the ELISA shows increased sensitivity Note The prevalence of the disease mainly is linked to the presence of compared to the IFAT (approximately 90% versus 50 – 70%). the vector (chiefly subtropical and tropical countries), but it can also The determination of antibody titres is unsuitable for therapy be encountered in animal shelters, where rhipicephalus often is able monitoring. Instead, serum protein electrophoresis and to survive the winter. The disease is transmitted by oral intake of an determination of CRP are recommendable. infected tick. 13.4.11 Neospora caninum 13.4.10 Leishmania A neurological disease in dogs whose pathogens were similar to toxoplasma but could Leishmaniosis is an infectious disease transmitted by insects. The vectors of Leishmania not be classified was first described in Norway in 1984. In 1988, a similar pathogen was are sand flies (Phlebotominae). Leishmania is taken up during the blood-sucking found in dogs in the USA and was named Neospora caninum. It was later determined process. The promastigote stages, which are infectious 6 – 12 days after the blood- that Neospora caninum was identical to the Norwegian pathogen. Neosporosis has sucking process, multiply in the sand fly. In Europe, the pathogen is Leishmania infantum. already been detected in many countries, it must therefore be assumed that it is spread South of the Bosporus and especially in North Africa, Leishmania tropica occurs worldwide. Natural infections have been found in dogs, cattle, horses, sheep, goats, red additionally. Other species of Leishmania have been described worldwide. The main deer and cats. Numerous other animals can be experimentally infected. infection areas in Europe are Spain, Portugal, Italy and Greece. Clinically, dogs and cattle are particularly severely affected. In the latter, at every stage of In Germany, naturally occurring sandflies have been found along the Rhine rift in Baden- gestation, the clinical picture is determined by abortions. In dogs, neurological signs are Württemberg, in Rhineland-Palatinate in the Kaiserslautern region, and in Saarbrücken. prominent: ascending paralysis of the hind legs with hyperextension are a typical finding, Foxes and possibly also small rodents are considered as pathogen reservoirs. but all limbs might be affected as well (tetraplegia). Other possible findings are dyspha- Infected animals can be asymptomatic for up to 7 years. The beginning of the disease gia, paralysis of the jaw, head tilt, muscle weakness, cardiac insufficiency and pneumo- is mostly characterised by lymphadenopathy, anaemia; in the cutaneous form of nia. Young, congenitally infected dogs show more severe signs, sometimes with sudden leishmaniosis, skin changes at the edges of the ears, the rhinarium and periorbital lesions deaths. Older dogs often show signs of disseminated infection with polyradiculitis, poly- are visible. myositis and possibly multiple organ involvement. Thus, in older dogs with neurological In chronic infections, the animals show reduced resilience, weight loss, signs, neosporosis should always be included in the differential diagnosis. However, due lymphadenopathy, scaly, non-itchy skin and eye changes. to the often high antibody prevalence in certain regions, it is assumed that only a small percentage of infected dogs actually develops a clinical disease. Leishmania – Pathogen Detection Neospora caninum – Pathogen Detection Material Swab without medium (conjunctiva), bone marrow, tissue (skin, lymph node, spleen), possibly EB Material Dog: faeces, CSF Method Realtime PCR, cytology, histology Cattle: abortion material, foetal tissue (brain, lung, liver, kidney) Species Dog, others on request Method Realtime PCR Duration 1 – 3 days Species Dog, cattle Duration 1 – 3 days Note Preferred method is PCR due to highest sensitivity. Neospora caninum – Antibodies Leishmania – Antibodies Material S, EP, HP 0.5 ml Material S 0.5 ml; IFAT also EP, HP Method IFAT, ELISA (cattle) Method IFAT, ELISA (only dog) Species Dog, cat, horse, cattle Species Dog, cat, horse Duration 1 day, cattle: 2 – 3 days Duration 1 day 226 227 2019/20 Infectious Diseases / Parasites 2019/20 Infectious Diseases / Parasites LABORATORY FOR CLINICAL DIAGNOSTICS 13.4.12 Nosema The mites burrow their tunnels into the horny layer of the skin. They prefer skin areas that are only sparsely haired, so they are often found on ears, elbows, lower abdomen Nosemosis is the most common disease of adult honey bees. It is caused by and ankles. If the disease spreads, larger areas of the body may be colonised. The unicellular, intracellular parasites, so-called microsporidia, which are closely related to main clinical sign is massive pruritus, which is often intensified by heat. Fungi. It is spread through spores that are viable for several years. Two species can be In pigs, the mites spread beginning from the inside of the pinna. Bovine sarcoptic differentiated: Nosema apis and Nosema ceranae, which can only be distinguished by mange especially affects the head and neck, but can also spread to the udder. Mange PCR, but differ in pathogenicity. The pathogens infect the midgut cells and thus lead causes loss of performance. to yellowish diarrhoea. Affected animals are often unable to fly and the abdomen is bloated. Many times, symptoms are rather non-specific, the bees are weak. Nosemosis Sarcoptes, Notoedres – Pathogen Detection is a multifactorial disorder, which means outbreaks of the disease only occur when Material Skin scraping (superficial, large-scale) there are other adverse conditions involved, such as cold, other illnesses, etc. Hence, Method (1) Microscopic evaluation nosemosis is potentially curable by resolving the other factors. Due to the resistance (2) Realtime PCR (Sarcoptes scabei var. canis) of the spores, it is often difficult to completely eliminate the pathogens. As with many Species (1) Dog, cat, farm animal, other species bee diseases, transmission occurs through the bees themselves (drifting or robbing) or (2) Dog, cat, rabbit, guinea pig, ferret, other canids and mustelidae through the beekeeper. Duration (1) 1 day (2) 1 – 3 days Nosema – Pathogen Detection Note Often, the infestation in dogs cannot be diagnosed by performing Material 30 – 40 dead bees a skin scraping. In this case, the diagnosis can only be made by Method (1) Microscopic evaluation antibody detection. (2) PCR (differentiation) In cats, localised infections are found in the head and neck area. Species Bees Zoonosis (pseudo scabies) Duration 1 – 2 days Note In case of a positive result, we recommend PCR differentiation Sarcoptes – Antibodies between Nosema apis and Nosema ceranae. Material S 0.5 ml Method ELISA Ostertagia Ø see Chapter 15.2 Species Dog Duration 1 day Note Seroconversion takes place 2 – 3 weeks post infection. Therapy 13.4.13 Sarcoptes monitoring cannot be carried out serologically due to persisting antibodies. Sarcoptes scabiei is the only species of the genus Sarcoptes. The Sarcoptes mites that are found in the different hosts are considered varieties of S. scabiei. The varieties are mostly host-specific, yet these itch mites are able to spread to other hosts, but usually 13.4.14 Toxoplasma do not settle there permanently. Sarcoptes scabiei varietas canis causes sarcoptic mange in dogs. Red foxes are Toxoplasma gondii is an obligate intracellular parasite which belongs to the class considered reservoir animals. Occasionally, the mite is also transmitted to ferrets, Coccidia. It is ubiquitous and causes clinical signs in all warm-blooded animals, rabbits, guinea pigs, cats and humans. including humans. Transmission occurs by direct contact between animals, but also indirectly via the More than 1 billion people worldwide have antibodies against toxoplasma. In addition contaminated environment. In dogs, indirect transmission seems to be gaining more to fever and cold-like symptoms, congenital infection during pregnancy is feared. and more importance. The whole developmental cycle of itch mites takes place on The intrauterine infection of the foetus occurs approximately 3 – 4 weeks after the first the host animal. In abraded skin material, the mites can survive up to 3 weeks, if the infection of a seronegative mother, when the placental barrier is crossed and placentitis environment is damp and cool. occurs. Miscarriages and severe neurological or ophthalmological diseases can occur 228 229 2019/20 Infectious Diseases / Parasites 2019/20 Infectious Diseases / Parasites LABORATORY FOR CLINICAL DIAGNOSTICS in the newborn. The cat as the definite host excretes oocysts for approx. 3 weeks, which 13.4.15 Trichomonads sporulate and become infectious after approx. 2 – 4 days (depending on temperature) (daily cleaning of the cat litter box!). Birds Another source of infection is meat contaminated with tissue cysts that has not been Trichomoniasis (also called canker or frounce) is a disease of the gastrointestinal tract, sufficiently cooked before consumption. However, the main source of infection is especially of the crop, which is caused by protozoa of the order Trichomonadida. In gardening, where oocysts may be absorbed via contaminated soil (aerosols). particular, the flagellates are transmitted through the crop milk or through contaminated Cats can also be intermediate hosts at the same time; they rarely fall ill, but the clinical drinking water. It is most notably pigeons and finches, but also budgerigars, cockatiels signs depend on where the tissue cysts are located. For example, hepatitis, cholangitis, and sometimes other parrots and canary birds that become infected. In pigeons, older dyspnoea may occur, and in case of CNS involvement, there may be ataxia, motor animals are often persistently infected, clinically inapparent carriers. Trichomonas deficits and epileptic seizures. Additionally, uveitis and chorioretinitis can occur. The gallinae is a pear-shaped flagellate of 5 to 18 µm in size that uses small lesions in the same signs can also be seen in dogs. mucous membrane to penetrate into the tissue and triggers the characteristic focal, In sheep and goats, about 10% of the abortions worldwide are attributed to T. gondii. yellowish tumours there. Occurrence of the disease is often associated with stress, In Germany, the authorities must be notified of the detection of Toxoplasma gondii vitamin deficiency or other illnesses and in some cases it can lead to the colonisation in cats, hares, rabbits, horses, ruminants, pigs and other mammals, especially those of inner organs such as the liver and the heart. Clinical signs often include regurgitation supplying food. of undigested food, but diarrhoea can be an indicator, too. In case of a longer duration of the disease, the animals lose weight and become apathetic. In young birds, the Toxoplasma gondii – Pathogen Detection mortality rate can be up to 40%. Material Cat: faeces (detection of excretion), CSF Dog, rabbit, guinea pig: CSF, tissue (e.g. brain) Reptiles Farm animals: abortion material, tissue (brain, heart, lung and others) If there are predisposing factors, trichomonads can contribute to enteritis in reptiles. Method (1) Realtime PCR (2) Faeces: microscopic evaluation after enrichment Trichomonads – Pathogen Detection Species Dog, cat, rabbit, guinea pig, farm animal, others on request Material Swab without medium (crop), lavage sample (crop) Duration (1) 1 – 3 days Method PCR (2) 1 day Species Birds Duration 1 – 3 days Toxoplasma – Antibodies Material S, EP, HP 0.5 ml Method IFAT 13.4.16 Tritrichomonas foetus Species Dog, cat, rabbit, guinea pig, farm animals, others on request Duration 1 day Tritrichomonas foetus is a protozoon belonging to the order Trichomonadidae. The trophozoite is characterised by three anterior flagella and one posterior flagellum. Note Detection of IgG (all species) and IgM (dog, cat, rabbit, guinea pig) However, similar to Giardia, these are only microscopically visible in fresh faecal samples. Cat: Usually, seronegative animals don´t excrete oocysts. Increased Transmission between cats occurs by faecal-oral route. Transmission between cattle or titres of IgM correlate with the excretion of oocysts. IgG antibodies pig to cat is not documented. indicate exposure and can point to clinical symptoms also in cats. Affected animals show typical large intestinal diarrhoea with frequent defaecation in small portions; admixtures of mucus and blood may occur. Tenesmus and uncontrolled defaecation are frequently observed. The general condition usually remains unaffected, increases in temperature are rare. T. foetus should always be considered as differential diagnosis in cats suffering from chronic, intermittent diarrhoea. In cattle, Tritrichomonas foetus causes bovine trichomoniasis, which is characterised by inflammation of the reproductive tract in cows leading to repeat breading and abortion. Bulls transmit the disease but display no clinical signs. In Germany, bovine trichomoniasis is notifiable upon suspicion. 230 231 2019/20 Infectious Diseases / Parasites 2019/20 Infectious Diseases / Parasites LABORATORY FOR CLINICAL DIAGNOSTICS Tritrichomonas foetus – Pathogen Detection Trypanosoma evansi Trypanosoma evansi (dog): North Africa, Middle East, Latin America, Asia Material Cat: faeces Cattle: swab without medium (cervix), preputial wash Method Realtime PCR Trypanosoma evansi – Pathogen Detection (Antigen) Species Cat, cattle, others on request Material EB 1 ml Duration 1 – 3 days Method Microscopic evaluation Note • Especially in cases of faecal Species Dog incontinence in cats, an Duration 1 day infection with Tritrichomonas foetus should be added to Trypanosoma evansi – Antibodies the list of differential Material S 0.5 ml diagnosis. Method CATT (card agglutination test for T. evansi) If anamnesis includes the Species Dog following information “patient Duration 1 day responded to therapy against Giardia, afterwards immediately recurrence”, this indicates Tritrichomonas foetus. • PCR is considered the most sensitive and specific method for the detection of Tritrichomonas foetus. As T. foetus is excreted intermittently, it is recommended to send a pooled faeces sample (collected over a period of 3 days) for analysis. 13.4.17 Trypanosoma Trypanosoma equiperdum Dourine is an infectious disease of equines which is transmitted during mating from horse to horse; course can be acute to chronic. Naïve reservoirs of Trypanosoma equiperdum are exclusively infected equines; mares and stallions host the pathogen in their genital secretions. Incubation time, severity and duration of disease vary markedly. Subclinical infections are possible; donkeys and mules seem to be more resistant. Clinically, animals show inflammatory changes of outer genital tract with depigmentation of mucosa up to peripheral-neurologic disturbances/paralysis. Especially in Asia and Africa still widespread; Central Europe is currently free from T. equiperdum. Trypanosma equiperdum (dourine) – Antibodies* Material S 1 ml Method CFT Species Horse Duration 5 days Note In Germany, dourine is an epizootic disease that is notifiable upon suspicion. 232 233 2019/20 Bacteriology/Mycology 2019/20 Bacteriology/Mycology LABORATORY FOR CLINICAL DIAGNOSTICS 14 Bacteriology/Mycology Blood Culture Parameter Pathogenic agents, 14.1 Smears/Puncture Fluids/Milk/Faeces aerobic + anaerobic Method Bacteriological culture (MALDI-TOF) Below, the most frequent requirements for general microbiology are listed. If indicated, aerobic + anaerobic special tests can be ordered separately (see Chapter 13.2, 13.3, 14.3, 14.4 and 16.1). Species Dog, cat, small mammals, birds, reptiles, large animals Duration 7 – 10 days The bacteriological examination detects aerobic pathogens. The detection of anaerobic Note • Please order blood culture flasks in advance (subject to a charge). pathogens needs to be requested separately. An exemption to this is blood culture, as it • Blood should be taken during a fever episode. is always incubated both aerobically and anaerobically. • Inoculation of the flask is done with 5 – 10 ml of blood. • It is recommended to send in 2 – 3 blood culture flasks (blood Because of the increase in multi-resistant pathogens such as MRSA (methicillin-resistant collection at different times, intervals of at least 1 hour). Staphylococcus aureus) or MRSP (methicillin-resistant Staphylococcus pseudointer • Storage and transport are uncooled. medius) as well as in Enterobacteriaceae of the ESBL strains (extended spectrum of • Bacteriaemia may occur physiologically in reptiles. beta-lactamases), a cultural examination and a subsequent antibiogram are almost indispensable in case of bacterial infections. Bronchial Lavage, Bronchial Secretion, Tracheal Secretion Parameter Pathogenic agents, aerobic Abscess Material Method Bacteriological culture (MALDI-TOF) and mycology Parameter Pathogenic agents, aerobic resp. anaerobic Species Dog, cat, small mammals, birds, reptiles, large animals Method Bacteriological culture (MALDI-TOF) and mycology Duration 2 – 3 days Species Dog, cat, small mammals, birds, reptiles, large animals Up to 7 days incl. mycology Duration 2 – 3 days Note • Please state if Bordetella bronchiseptica is suspected as special Up to 7 days incl. mycology culture media are required! Anaerobic approx. 1 week • For microbiological examination, lavage fluid should be sent by Note Split the abscess cavity and swab the inside of the abscess use of a swab with transport medium. membrane; requesting an anaerobic bacteriological examination is recommended, too. Cerebrospinal Fluid Parameter Pathogenic agents, aerobic resp. anaerobic Aspirates (from primarily sterile body cavities) Method Bacteriological culture (MALDI-TOF) and mycology Parameter Pathogenic agents, aerobic resp. anaerobic Species Dog, cat, small mammals and large animals Method Bacteriological culture (MALDI-TOF) and mycology Duration 2 – 3 days Species Dog, cat, small mammals, birds, reptiles, large animals Up to 7 days incl. mycology Duration 2 – 3 days Up to 7 days incl. mycology Ear Smear Anaerobic approx. 1 week Parameter Pathogenic agents, aerobic Note • In case of punctures from primarily sterile body cavities, the Method (1) Bacteriological culture (MALDI-TOF) and mycology sample material should not be cooled. (2) Parasitology • If testing for actinomycetes and Nocardia is required, it needs to Species Dog, cat, small mammals, large animals be requested separately. Duration (1) 2 – 3 days (2) 1 day 234 235 2019/20 Bacteriology/Mycology 2019/20 Bacteriology/Mycology LABORATORY FOR CLINICAL DIAGNOSTICS Note • Please send in a swab with medium for the cultural examination. Wound Swab If a parasitological examination is also requested, a swab without medium must be sent in additionally. Parameter Pathogenic agents, aerobic resp. anaerobic • An antimycogram for Malassezia spp. is only performed on Method Bacteriological culture (MALDI-TOF) and mycology special request. Species Dog, cat, small mammals, birds, reptiles, large animals Duration 2 – 3 days Up to 7 days incl. mycology Faeces Anaerobic approx. 1 week Parameter Aerobic facultative and obligate pathogenic germs and fungi Method Bacteriological culture (MALDI-TOF) and mycology Species Dog, cat, small mammals, birds, large animals Duration 2 – 3 days 14.2 Skin/Hair/Feathers Up to 7 days incl. mycology Skin Swabs Note Diagnostic findings and significance of bacteriological faecal examination see Chapter 16.1. Parameter (1) Pathogenic agents, aerobic (2) Dermatophytes, yeasts, moulds Method (1) Bacteriological culture (MALDI-TOF) Milk (2) Culture, mycological Parameter Pathogenic agents, aerobic, including determination of germ count Species Dog, cat, small mammals, birds, reptiles, large animals and inhibitor test Duration (1) 2 – 3 days Method Bacteriological culture (MALDI-TOF), mycology (2) Up to 4 weeks Species Cow, sheep, goat, horse Note If an additional parasitological examination is required, hairs or a Duration 2 – 3 days skin scraping need to be submitted. Note Examination of both 1/4 and 4/4 milk samples is possible. Skin, Danders, Hairs, Feathers Smears (nose, pharynx, urethra, vagina etc.) Parameter (1) Pathogenic agents, aerobic Parameter Pathogenic agents, aerobic resp. anaerobic (2) Dermatophytes, yeasts, moulds Method Bacteriological culture (MALDI-TOF) and mycology (3) Parasites Species Dog, cat, small mammals, bird, reptiles, large animals Method (1) Bacteriological culture (MALDI-TOF) Duration 2 – 3 days (2) Culture, mycological + paraffin oil preparation Up to 7 days incl. mycology (3) Parasitology: paraffin oil preparation Note For detection by culture a swab with medium is required. A detection Species Dog, cat, small mammals, birds, reptiles, large animals by culture from purulent material is often difficult as the bacteria are Duration (1) 2 – 3 days pre-damaged and therefore difficult to breed. (2) Up to 4 weeks (3) 1 day Urine Bacteriological Culture Note In case of pathogenic yeasts and dermatophytes, an antimycogram can be performed on special request. Material Pathogenic agents, aerobic, including determination of germ content It is absolutely necessary to collect samples from the edge of the (semi-quantitative) skin changes. Method Bacteriological culture (MALDI-TOF) Species Dog, cat, small mammals, large animals Duration 2 – 3 days Note Submission of urine and swab of urine (with medium) is optimal. 236 237 2019/20 Bacteriology/Mycology 2019/20 Bacteriology/Mycology LABORATORY FOR CLINICAL DIAGNOSTICS Trichogram/Pennogram Taylorella equigenitalis (CEM) Parameter Current condition of coat/plumage Material Cervical or clitoral swab (mare) Method Microscopy Shaft swab, urethral swab or glans penis swab (stallion) Species Dog, cat, small mammals, birds, reptiles (1) Swab with medium (Amies, ≤ 48 h) Duration 1 day (2) Swab with medium Note In skin patients, a trichogram serves as an additional method for Method (1) Bacteriological culture (MALDI-TOF) diagnostic assessment. It cannot replace histology, bacteriological, (2) Realtime PCR mycological and parasitological examinations, cytology and other Species Horse tests, such as the determination of clinical-chemical parameters or Duration (1) 7 days, for export up to 21 days hormone assays, but it can provide very valuable information. (2) 1 – 3 days Trichograms are particularly suitable in the diagnosis of cats with Note • Culture: For the export to Canada, the incubation time is 14 days, hair loss that have no apparent itching and for which alopecia sine to Norway 21 days, otherwise 7 days causa is already clinically suspected. A trichogram can also provide • Detection by PCR is offered as a single test for one sample or as valuable diagnostic information in case of colour mutant alopecia. a profile for 2 samples (mare) or 3 samples (stallion). • Obligation to notify the authorities upon diagnosis. 14.3 Bacteriological Examination Horse 14.4 Testing for Specific Infectious Agents Reproductive Fitness Material Mare: swab with medium (cervix) Actinomyces Stallion: swab with medium (penile sheath, urethra, glans penis) Material Aspirates, swab with medium etc. Parameter: Pathogenic agents, aerobic Method Bacteriological culture Method Bacteriological culture (MALDI-TOF) and if necessary mycology Species Dog, cat, small mammals and large animals Species Horse Duration Approx. 8 days Duration 2 – 3 days, mycology: 7 days Note This examination is offered as service „Nocardia/Actinomyces“. Note This examination can be ordered with or without mycological examination. Bordetella bronchiseptica (aerobic) Furthermore, we offer a combination of cultural and mycological examination as well as pathological examination of 1 – 3 endometrial Material (1) Swab with medium (Amies) (nose, pharynx), bronchial secretion biopsies for mares. (2) Swab without medium, bronchial secretion, bronchoalveolar lavage Method (1) Bacteriological culture (MALDI-TOF) Streptococcus equi (2) Realtime PCR Material (1) Swab with medium (nose, abscess, lymph node), BAL, TBS Species Dog, cat, rabbit, cattle, sheep, goat, pig, other species (2) Swab without medium (nose), lavage sample (bronchoalveolar Duration (1) 2 – 3 days lavage, tracheobronchial secretion), tissue (lymph node) (2) 1 – 3 days Method (1) Culture Note When requesting a bacteriological examination, please indicate (2) Realtime PCR clearly on the submission form that Bordetella bronchiseptica should Species Horse be tested, as special culture media are required. Duration 1 – 3 days Note The two subspecies Streptococcus equi equi and Streptococcus equi zooepidemicus are detected and differentiated by cultural cultivation as well as by PCR. 238 239 2019/20 Bacteriology/Mycology 2019/20 Bacteriology/Mycology LABORATORY FOR CLINICAL DIAGNOSTICS Dermatophilus congolensis 14.5 Sensitivity Testing Material Scurfs Method Slide - quick staining We perform all antibiograms using microdilution technique according to CLSI standard. Species Horse, ruminants Duration 1 day Antibiogram (aerobes) Note Hair alone is not sufficient as sample material. There are species-specific standard programs (see notation). An antibiogram is invoiced as a fixed price per bacteriological culture, even if several antibiograms need ESBL – Pathogen Detection to be performed. Material Swab with medium, faeces Duration 2 – 3 days Method Culture Species Dog, cat, horse, cattle, other species Note Species-specific antibiograms: Duration 3 – 4 days • small animals • rabbits and rodents Note • Extended-spectrum β-lactamase (ESBL)-producing Enterobacte- • birds riaceae such as E. coli, Klebsiella sp. and Proteus sp. are called • reptiles and amphibians ESBL. Due to the specific lactamase with extended spectrum • large animals of activity, the bacteria are resistant to β-lactam antibiotics and • fish cephalosporins (also to 3rd and 4th generation cephalosporins). The property of ESBL formation is encoded on easily transferable genetic segments and, during reproduction, can be transferred Antibiogram (anaerobes) from one bacterial generation to the next (vertical transfer) or We can also perform an antibiogram if anaerobes are detected. Only antibiotics with exchanged between bacteria (horizontal transfer). potential efficacy against anaerobes are tested. • Only after prior bacteriological examination Duration 5 – 7 days after identification of the pathogen Nocardia Material Aspirates, swab with medium etc. Antibiogram Extended Method Bacteriological culture The complexity of an extended antibiogram depends on the respective species. Species Dog, cat, small mammals and large animals Duration Approx. 8 days Duration 2 – 3 days Note This examination includes the detection of Actinomyces. Antimycogram Paenibacillus larvae (aerobic) We can perform an antimycogram if yeasts or dermatophytes have been cultured. This Material Honey and wax sample needs to be requested, though. We store the cultivated yeasts and filamentous fungi for Method Cultural, bacteriological (MALDI-TOF) one week. Species Bees Duration 3 – 5 days Duration 28 – 54 days Note • Two tablespoons of feed honey from the honey dome should be scratched from a central brood comb, packed into a freezer bag and sent in. • American (malignant) foulbrood is an epizootic disease that is notifiable upon suspicion. 240 241 2019/20 Bacteriology/Mycology 2019/20 Parasitology LABORATORY FOR CLINICAL DIAGNOSTICS 14.6 Additional sensitivity testing 15 Parasitology Aromatogram 15.1 Parasitological Examination – Faeces The aromatogram is an in vitro test for testing the sensitivity of bacteria and yeasts/ Malassezia to various essential oils. The procedure is based on the principle of the agar Parasite Profile Ø see also Chapter 2.1.6, 2.2.3, 2.3.3, 16.1.1 diffusion test (disk diffusion test). (Faecal Profiles Small Animals and Horse) The in vitro efficacy of essential oils is classified into 4 categories: from ineffective to Below, the most frequent requirements for parasitological faecal examination are listed. slightly, moderately and highly effective. Currently, 13 different essential oils can be For the enrichment by flotation or sedimentation, we need a cherry-sized amount of tested. faeces, if possible a 3-day pooled faecal sample. For serological detection by EIA, a pea-sized amount is usually sufficient. Duration Bacteria: 2 days Yeasts: up to 7 days Egg Count: Modified McMaster Method Counting of worm eggs is done using a counting chamber after enrichment by flotation. This method is primarily used in horses, sheep and other farm animals to carry out targeted or selective deworming, in which individual deworming is only carried out if there are > 200 eggs per gram of faeces. If all animals of a population that have a worm infestation are dewormed, only resistant worms survive. However, if only animals that have a more severe worm infestation are dewormed, an untreated worm population is also found in the animal population, which reduces the selective advantage of resistant worms and thus counteracts the further increase in resistance. Duration 1 day Egg Count Reduction Test 2nd test: Within the concept of selective deworming (see above), further individual samples are taken 10 – 14 days after therapy. If high egg counts are still present, this finding is suspicious of anthelmintic resistance. Even if deworming does not take place selectively at the level of the individual animal but, for example, of groups of animals, regular monitoring of the effectiveness of anthel- mintics by means of an egg count reduction test is recommended. Duration 1 day Endoparasites Material Faeces Method Microscopy after enrichment by flotation and SAF (Sodium Acetate Acetic Acid Formalin Solution, special sedimentation method) Species All Duration 1 day 242 243 2019/20 Parasitology 2019/20 Parasitology LABORATORY FOR CLINICAL DIAGNOSTICS Note • Parasite or protozoa eggs are shed intermittently. In suspicious Fasciola Hepatica – Antibody Detection cases, the test has to be repeated (it is best to send 3 consecutive samples) Material S, HP, (tank) milk 0.5 ml • Flotation and SAF techniques are used for each sample. Method EIA Species Cattle Duration 3 – 4 days Endoparasites (Reptiles) Material Faeces Ostertagia Ostertagi – Antibody Detection Method Microscopy after enrichment by SAF (Sodium Acetate Acetic Acid Formalin Solution, special sedimentation method), Ziehl-Neelsen Material Milk, tank milk 0.5 ml staining, fresh preparation Method ELISA Species Reptiles Species Cattle Duration 1 day Duration 3 – 4 days Note Parasite eggs or protozoa are only shed intermittently, so on suspicion, the test needs to be repeated. 15.3 Parasitological Examination – Skin Lungworm Larvae (Baermann Test) Skin Material Faeces Method Baermann examination Material Skin scraping Species Dog, cat, small mammals, large animals Method Microscopy Duration 2 days Species Dog, cat, small mammals, large animals Duration 1 day Note • Baermann examination is indicated in cases of chronical cough and dyspnoea (because of lungworm larvae infections). Note The detection of ectoparasites is done by examination of skin • Angiostrongylus vasorum can be detected with a Baermann scrapings. These should be placed into shipping containers before test as well as by PCR from blood or bronchoalveolar lavage (see they are sent in. Chapter 13.4). 15.2 Testing for Specific Parasitic or Protozoa Infections Echinococcus – Pathogen Detection* Material Faeces Method (1) EIA (antigen detection) (2) PCR Species Dog, cat, fox Duration Approx. 1 week Note Detection by EIA or PCR can reveal infections with E. granulosus and E. multilocularis, while microscopy after enrichment is often only able to detect non-differentiable Taenia eggs. Echinococcosis is a notifiable disease. 244 245 2019/20 Tests for Indigestion and Diarrhoea 2019/20 Tests for Indigestion and Diarrhoea LABORATORY FOR CLINICAL DIAGNOSTICS 16 Tests for Indigestion and Diarrhoea Faecal Profile BARF Salmonella including enrichment, Yersinia including enrichment, campylobacter, listeria, 16.1 Bacteriological Examination endoparasites The physiological intestinal flora consists of numerous bacterial species that live Duration 2 – 3 days (except Yersinia: 21 days) together with the host in a complex symbiotic ecosystem. Shortly after birth and the Note See also biochemistry profile BARF (Chapter 2.1.1). suckling phase, the gastrointestinal flora is established and remains largely stable for the rest of the life. Faecal Profile Pathogenic Bacteria (dog) However, within the intestinal tract, there are considerable differences in distribution. Salmonella including enrichment, Yersinia including enrichment, campylobacter, entero- While pathogen counts in the duodenum and the jejunum are rather low due to the pathogenic E. coli incl. virulence factors (STa, stx1, stx2, eae) influence of gastric acid, bile and pancreatic enzymes as well as the present mucosal defence systems, they massively increase in the ileocecal area and reach their highest Duration 2 – 3 days (except Yersinia: 21 days) concentration in the large intestine. The number of anaerobes and facultative anaerobic pathogens is 1,000 to 10,000 times higher than the number of aerobic microflora. The highest concentrations are reached by Bacteroides spp., lactobacilli and bifidobacteria Large Faecal Profile (small animals) as well as Enterobacteriaceae. Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria including enrichment for salmonella, Clostridium perfringens enterotoxin, Clostridium The advantage of culture examination is that targeted antibiotic treatment can be per- difficile toxin A and B, endoparasites formed based on the antibiogram. This is necessary because of the MRSA/MRSP/ESBL problem. ESBL producers have a specific β-lactamase and are resistant to almost all Parasites Profile Cat Ø see Chapter 2.1.6 β-lactam antibiotics as well as to 3rd and 4th generation cephalosporins. Such patho- gens are found, for example, in E. coli, Proteus sp., Klebsiella, Enterobacter, Serratia, Puppy Faecal Profile Citrobacter and Salmonella. Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria including enrichment for salmonella, gas producers, endoparasites, parvovirus 16.1.1 Faecal Profiles Small Faecal Profile The faeces tube should be ¾ full, if possible. An aerobe bacteriological and possibly mycological examination, including enrichment, is performed for salmonella and shigella. Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria Pathogen differentiation is done by MALDI-TOF. Unless otherwise stated, the test duration including enrichment for salmonella, gas producers is 2 – 3 days. If required, serological pathogen differentiation (e.g. salmonella) and an antibiogram, which are subject to a charge, can be performed additionally. Dog and Cat – Virological Faecal Profiles Virological Faecal Profile Dog and Cat Parvovirus, rotavirus, coronavirus Combined Faecal Profile (dog/cat) Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria including enrichment for salmonella, gas producers, endoparasites as well as Giardia Dog and Cat – Faecal Profiles PCR sp. antigen-EIA and Cryptosporidia antigen-EIA Diarrhoea, Human Pathogenic Causes Salmonella, Yersinia enterocolitica, Campylobacter jejuni 246 247 2019/20 Tests for Indigestion and Diarrhoea 2019/20 Tests for Indigestion and Diarrhoea LABORATORY FOR CLINICAL DIAGNOSTICS Diarrhoea (dog) Small Faecal Profile Coronavirus, parvovirus, circovirus, giardia, cryptosporidia Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria including enrichment for salmonella, gas producers Diarrhoea (cat) Coronavirus, Tritrichomonas foetus, giardia, parvovirus, cryptosporidia Ruminants Bovine Faecal Profile Small Mammals, Birds and Reptiles Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria Avian Faecal Profile + Endoparasites including enrichment for salmonella, endoparasites; This profile also includes the detection of M. avium ssp. Paratuberculosis by PCR. Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria including enrichment for salmonella, endoparasites Calf Faecal Profile – Large Note This profile is also available without the detection of endoparasites. Bacteriology (aerobic) and mycology including enrichment for salmonella, endoparasites, rotavirus, coronavirus Ferret Faecal Profile Note If salmonella or E. coli are detected, they will be serologically typed Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria as an additional service (subject to a charge). including enrichment for salmonella, endoparasites, Giardia sp. antigen EIA Calf Faecal Profile (EIA) Pigeon Faecal Profile Detection of rota- and coronavirus, E. coli K99, cryptosporidia Salmonella incl. enrichment, endoparasites (incl. coccidia) Note The advantage of ELISA is the speed of scanning. Reptile Faecal Profile Faecal Profile Alpaca, Llama (camelids) Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria including enrichment for salmonella Bacteriology (aerobic) and mycology including enrichment for salmonella, endoparasites, rotavirus, coronavirus, Clostridium perfringens enterotoxin Rodent Faecal Profile + Endoparasites Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria Pig including enrichment for salmonella, endoparasites Piglet Faecal Profile Bacteriology (aerobic) and mycology including enrichment for salmonella, endoparasites, Horse rotavirus, coronavirus, Clostridium perfringens enterotoxin Foal Faecal Profile Note If salmonella or E. coli are detected, they will be serologically typed as an additional service (subject to a charge). Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria including enrichment for salmonella, gas producers, rotavirus, Clostridium perfringens enterotoxin, endoparasites Porcine Faecal Profile Bacteriology (aerobic) and mycology including enrichment for salmonella Large Faecal Profile This profile also includes the detection of Lawsonia intracellularis by PCR. Bacteriology (aerobic) and mycology, obligate and facultative pathogenic bacteria including enrichment for salmonella, gas producers, Clostridium perfringens enterotoxin, Clostridium difficile toxin A and B, endoparasites 248 249 2019/20 Tests for Indigestion and Diarrhoea 2019/20 Tests for Indigestion and Diarrhoea LABORATORY FOR CLINICAL DIAGNOSTICS 16.1.2 Testing for Specific Indigestion/Diarrhoea Pathogens Note • Examination is specifically indicated if colitis-like symptoms occur. (Bacteria) • Faeces sample of 2 cm of diameter or larger is required. • In carnivores, Clostridium perfringens enterotoxin can cause diarrhoea and vomiting of varying severity; enterotoxaemia is rare. Campylobacter – Pathogen Detection Toxin formation is induced by antibiotic administration, stress, co- Material Faeces, swab with medium (intestine, cloaca) infections or especially by an unbalanced diet rich in proteins and Method (1) Culture examination (MALDI-TOF) connective tissue. (2) Realtime PCR (only detection of Campylobacter jejuni) Species No limitations known E. coli eae Gene Duration (1) 2 – 3 days (2) 1 – 3 days Material Faeces Method PCR after prior detection of E. coli Note • A faecal sample with a diameter of at least 2 cm is required, Species Calf, piglet otherwise use swab with medium; for PCR: swab without medium Duration 3 days • A combined cultural detection of Campylobacter and Yersinia is also available. Note The eae-gene is a pathogenicity factor of E. coli. The eae-gene (E. • Resistances are common; therapy should therefore only be coli attaching and effacing) encodes the production of intimin, which carried out after an antibiogram has been performed. Preparation permits E. coli to attach itself to the intestinal cells. of an antibiogram is only possible after cultural examination. • In cattle, C. jejuni causes diarrhoea and mastitis. C. coli is Helicobacter spp. detected in case of mild diarrhoea in pigs and in case of abortion Material Vomit, gastric lavage, gastric biopsy, sheep: abortion material in sheep. Method PCR • Campylobacter of the species C. jejuni, C. coli, C. lani and Species Dog, cat, ferret, sheep C. upsalensis are classified as thermophilic Campylobacter. Duration 1 – 3 days Campylobacter is one of the most common causes of bacterial diarrhoea in humans (mainly from poultry products). Note • Positive PCR results from faecal samples do not necessarily • Campylobacteriosis (thermophilic Campylobacter) is a notifiable indicate involvement of the stomach (gastritis, stomach ulcer disease in dogs, cats, ruminants and poultry. etc.), as PCR also detects intestinal Helicobacter spp. For this diagnostic task, stomach biopsies or vomitus are recommended. • PCR accurately detects at least H. pylori, H. felis, H. bizzozeroni, Clostridium difficile Toxin A and B H. hepaticus, H. bilis, H. heilmannii, H. muridarum, H. mustelae, Material Faeces H. cinaedi, H. canis and Flexispira rappini. Method ELISA • Flexispira rappini is also assigned to the genus Helicobacter and Species No limitations known is associated with the occurrence of abortions in sheep. Duration 2 days Note • Examination is specifically indicated if colitis-like symptoms occur. Macrorhabdus ornithogaster • A faecal sample with a diameter of at least 2 cm is required. Material Faeces, proventriculus, smear on slide Method Stain, microscopy Species Birds Clostridium perfringens Enterotoxin Duration 1 day Material Faeces Method ELISA Species No limitations known Duration 2 days 250 251 2019/20 Tests for Indigestion and Diarrhoea 2019/20 Tests for Indigestion and Diarrhoea LABORATORY FOR CLINICAL DIAGNOSTICS Mycobacteria (Microscopic Detection of Acid-fast Rods) 16.2.2 Testing for Specific Indigestion/Diarrhoea Pathogens Material Faeces, smear on glass slide (viruses) Method Ziehl-Neelsen stain Species No limitations known Coronaviruses – Pathogen Detection Duration 1 day Material Faeces, pig: also tissue (e.g. intestine) Note A faecal sample with a diameter of at least 2 cm is required. Method Realtime PCR Species Dog, cat, horse, cattle, pig Duration 1 – 3 days Salmonella and Shigella Note A faecal sample with a diameter of at least 2 cm is required. Material Faeces (Swab with medium: intestine, cloaca) Method Culture including enrichment, MALDI-TOF Parvovirus – Pathogen Detection Species No limitations known Material Dog, cat: faeces, EB 0.2 ml Duration 2 – 3 days Pig: tissue (e.g. abortion material, lung, liver), sperm, EB Note A faecal sample with a diameter of at least 2 cm is required, in Method Realtime PCR exceptional cases a swab with transport medium can be used too. Species Dog, cat, pig Duration 1 – 3 days Note • PCR can be positive up to four weeks after vaccination with live Yersinia vaccine. Material (1) Faeces (swab with medium) • In dogs, differentiation between vaccine strain (CPV 2) and field (2) Faeces, swab without medium (rectum) strains (CPV 2a, 2b, 2c) is possible on request. Please note that Method (1) Culture examination including enrichment, MALDI-TOF when using parvovirus vaccines containing CPV 2b as vaccine (2) Realtime PCR (only Y. enterocolitica) antigen (e.g. Virbagen Puppy 2b) we cannot differentiate between Species No limitations known vaccine strain and field strain. Duration (1) Up to 3 weeks • Direct detection of parvoviruses in the blood is possible approx. (2) 1 – 3 days 1 – 5 days after infection. Note • A faecal sample of 2 cm of diameter or larger is required. In exceptional cases, a swab with transport medium can be used Parvovirus – Antigen too (culture). Material Faeces • Detection by culture is also offered as a combined service Method EIA together with the detection of Campylobacter. Species Dog, cat Duration 1 day Note A faecal sample with a diameter of at least 2 cm is required. The 16.2 Virological Examination test may yield positive results 5 – 12 days after vaccination with live vaccine. 16.2.1 Faecal Profiles – Virology Rotavirus – Antigen Material Faeces Virological Faecal Profile (dog, cat) Method EIA Parvovirus, rotavirus, coronavirus Species Dog, cat, horse, cattle Duration 1 day Note A faecal sample of 2 cm of diameter or larger is required. 252 253 2019/20 Tests for Indigestion and Diarrhoea 2019/20 Tests for Indigestion and Diarrhoea LABORATORY FOR CLINICAL DIAGNOSTICS 16.3 Diagnostics of Maldigestion/Malabsorption Particle Size Material Faeces Bile Acids Method Measurement Material Faeces Species Horse Method ELISA Duration 1 day Species Dog, cat Note Particle size provides information on the insufficient disintegration of Duration Up to 7 days the feed components. Note A bacterial overgrowth or a post-OP slow intestinal flow can produce Dental examination should follow. Furthermore, the ration should be diarrhoea, caused by the presence of bile acids in faeces. The symp- checked with regard to the amount of components that are difficult toms are aqueous diarrhoea or steatorrhoea. to digest (e.g. excessive feeding of straw) and to structure (e.g. sufficient fibre length). Chymotrypsin Sand Content Material Faeces Method ELISA Material Faeces Species Cat Method Wash out and measurement Duration 2 – 5 days Species Horse Duration 1 day Note In case of exocrine pancreatic insufficiency. Levels of Chymotrypsin depend on the gastrointestinal passage Note Recurrent digestion problems can be caused by an excess of sand time. Therefore, measurement should not be performed during an in the intestines or fodder rich in sand. acute phase of diarrhoea. Microscopical Nutritive Digestion 16.4 Determination of an Inflammatory Exudative Material Faeces Process Method Microscopy Species Dog, cat A faecal sample with a diameter of at least 2 cm is required for these examinations. Duration 1 day Note This is a semiquantitative analysis of those parts of the aliments that Albumin/Globulin remain undigested in the faeces. As it depends on the composition Material Faeces of diet, it is not possible to point out a malabsorption-maldigestion Method Gel electrophoresis problem. Species Dog, cat Test frequency 1 x/week Pancreas Elastase Note Detection of inflammatory exudative colitis. Material Faeces Method EIA Calprotectin Species Dog Duration 2 days Material Faeces Method Turbidimetry Note With the pancreas elastase test we check the pancreas exocrine Species Dog function in dogs. Duration 1 day The elastase is specific for the pancreas, stable in the intestine, and the test results are not altered by a substitution therapy. Note Biomarker used to diagnose suspected inflammatory bowel disease, IBD. 254 255 2019/20 Tests for Indigestion and Diarrhoea 2019/20 Autovaccine/Herd-specific Vaccine LABORATORY FOR CLINICAL DIAGNOSTICS Chemical Detection of Blood 17 Autovaccine/Herd-specific Vaccine Material Faeces Method Chemical (modified Guajak test) Chronic recurrent infections in pets and domestic animals are not uncommon and Species No limitations known often lead to an increased resistance of the pathogens involved because of prolonged Duration 1 day antibiotic therapies. When considering alternative methods of treatment, autovaccines are often mentioned, which have successfully been used in human and veterinary Note To avoid false-positive results, carnivores should be fed no meat for medicine for quite some time. These are customised vaccines that are produced for 3 – 4 days. each individual animal from its own aerobic bacterial flora. α-1-Antitrypsin Production of an autovaccine is always based on a microbiological examination Material Faeces during which the causative pathogen is isolated. It is multiplied in pure culture and Method ELISA then inactivated. The specific composition of the vaccine, such as concentration of Species Dog germs, method and amount of application, depends on the preliminary report and the Test frequency 1 x/week animal species. In general, you have to differentiate between the local and the systemic application of autovaccines. Injection vaccines that are given subcutaneously are Note Detection of protein-losing-enteropathy (PLE). especially recommended in case of skin infections, particularly with Staphylococcus pseudointermedius, for respiratory diseases, otitis, etc. If the nasopharyngeal area is affected, the autovaccine can also be prepared for use as an aerosol. Infections of the urogenital tract can be treated more effectively with combination vaccines, that are either used orally or systemically. Oral vaccines are administered in all gastrointestinal infections associated with chronic diarrhoea. The aim of the treatment is to sensitise the immune system against the causative pathogen or against its metabolites and to stimulate it to produce specific antibodies. Thus, the present infection is fought off and reinfection is prevented. The production of secretory IgA results in the mucosal barrier being “sealed”. Regarding the problem of watery stools in horses, very good results have been achieved with the autovaccine. Please note: We need a prescription from your veterinary practice for the preparation of this product! Autovaccine (Oral Vaccine) for Chronic Diarrhoea Material Faeces Method Bacteriological culture of the aerobic germs, then inactivation of the pathogens and subsequent production of the autovaccine solution. Species Dog, cat, rabbit, rodents, ferret, parrot, budgerigar, mynah, hawk, pigeon, ostrich, tortoise, snake, horse, goat, llama, alpaca, koi, gibbon, orang-utan, gorilla, marmosettes, giant panda, lion, tiger, elephant, tapir, elk, giraffe, Bactrian camel, kangaroo (Applies only to non-food producing animals.) 256 257 2019/20 Autovaccine/Herd-specific Vaccine 2019/20 Pathology LABORATORY FOR CLINICAL DIAGNOSTICS Duration 3 weeks 18 Pathology Note Is administered as an oral vaccine. Please note: We need a prescription from your veterinary practice for the preparation of this product! 18.1 Histopathology Autovaccine (Injection/Inhalation Vaccines) Endometrial Biopsy (mare) Material Swabs with medium, urine, hair etc. Material 3 tissue samples approx. 1.0 x 1.0 x 0.5 cm (1 x corpus, 2 x cornua Method Bacteriological culture of the aerobic germs, then inactivation of the uteri), formalin fixed (4% neutral buffered formaldehyde 10% pathogens and subsequent production of the autovaccine solution. formalin) Species Dog, cat, rabbit, rodents, ferret, Method Microscopy (standard und special stainings) ≙ parrot, budgerigar, mynah, hawk, pigeon, ostrich, Duration 2 days tortoise, snake, Note • For the following clinical issues: breeding suitability test, barren horse, mare, abortion etc. goat, llama, alpaca, • Histological diagnosis of endometritis, endometrosis, koi, angiopathies, (pathological) inactivity, lymphatic lacunae, false gibbon, orang-utan, gorilla, marmosettes, giant panda, lion, tiger, differentiations etc. elephant, tapir, elk, giraffe, Bactrian camel, kangaroo • Fertility prognosis (categorisation according to Kenney & Doig (Applies only to non-food producing animals.) 1986, mod. according to Schoon et al. 1992) Duration 3 weeks • Uterine biopsy can also be requested as a combined service Note The following can be produced: together with breeding hygiene testing and mycological • Injection vaccines (e.g. in case of pyoderma etc.) examination. • Combination vaccines (e.g. in case of vaginitis and cystitis) • Inhalation vaccines in case of respiratory diseases Histopathology Please note: We need a prescription from your veterinary practice Material • Formalin-fixed (10%) tissue samples (fixation in 4% neutral for the preparation of this product! buffered formaldehyde 10% formalin) • For dermatologic issues use skin punches (≥ 0.6 cm). Herd-specific Vaccines and Autovaccines: Indications in Small Animals Method Microscopy (standard und≙ special stainings) Autovaccines against Administration Sample material Duration 2 days skin infections (particularly Note Fill in the submission form Pathology. Staphylococcus pseudointermedius), injection vaccines swab samples respiratory diseases, otitis, and others chronic recurrent infections inhalation vaccines nasal swab in the ENT area (aerosol autovaccines) 18.2 Immunohistology gastrointestinal infections with chronic diarrhoea oral vaccine faecal swab/faeces chronic recurrent infections in the combination vaccines swab samples (vagina), Immunohistology urogenital region (per os and injection) urine Material Formalin-fixed and/or paraffin-embedded tissue samples Method Microscopy (labelling with specific antibodies) Duration Up to 1 week Note Examination only after histopathological diagnosis, e.g. tumour diagnosis. • CD3/CD20 in lymphoma diagnosis • c-kit expression pattern in mast cell tumours 258 259 2019/20 Pathology 2019/20 Pathology LABORATORY FOR CLINICAL DIAGNOSTICS • Ki-67 antigen as proliferation marker included in the overall diagnosis (summary probability diagnosis). • Cox-2, prostaglandin synthesis enzyme, in tumours possibly an Cell material can then be obtained directly from the cytological indicator of the effectiveness of inhibitors (NSAIDs) smear or the histological paraffin block. • Differentiation between epithelial/spindle cell/round cell/ melanocytic tumours Infection diagnosis: 18.5 BRAF Mutation • FIP virus, parvovirus, FeLV, toxoplasmoses, feline herpes virus BRAF Mutation (transitional cell carcinoma) 18.3 Cytology Material Formalin fixed tissue or 5 – 10 ml urine or smears Method Sequencing Species Dog Cytological Examination Duration 3 days Material Aspirate, air dried smears on slides after puncture, impression smears Note Transitional cell carcinomas (TCC) are the most common malignant or fine needle aspiration, vaginal cytology tumours of the canine urinary tract. Mochizuki et al. (PLoS ONE Method Microscopy (standard and special stainings) 2015a, 10(6):e0129534; PLoS ONE 2015b, 10(12):e0144170) Duration 1 day identified the BRAF variant V595E in approximately 85% of canine Note Please send native liquids (aspirates, excreta, secretion) for additional TCC cases by DNA sequencing of TCC tumour cells as well as in clinical chemistry in untreated tubes (can be used for bacteriology prostate carcinoma. as well, if necessary) and additionally in EDTA tubes (superior cell This BRAF variant V595E analysis is a non-invasive and simple tool morphology). used to confirm the diagnosis in cases where TCC is suspected. However, it is important to note that only a positive identification of the mutation is valuable, since the variant could never be demonstrated in animals with cystitis, polyp or other urinary bladder neoplasms. 18.4 Lymphocyte Clonality (PARR) The absence of the BRAF mutation may be due to the fact that the tumour is not caused by a BRAF mutation or that the submitted Molecular Biological Clonality Analysis of Lymphocytes material did not contain mutated cells. In addition, the tumour might Material Tissue, cytological smears, lymphoid material not be a TCC, but a tumour of other origin, a polyp or cystitis. Method PARR (Polymerase Chain Reaction for Antigen Receptor Rearrangements) Species Dog, cat 18.6 Differentiation Exsudate/Transudate Duration 2 – 4 days Note The examination serves to confirm a suspected diagnosis or, if Parameter Transudate Exsudate Modified Transudate negative, makes a lymphoma diagnosis less likely. In positive cases, a differentiation between B- and T-cell lymphomas Colour Slightly yellow Bloody, purulent Variable is done at the same time (with exceptions). Transparency Clear Mostly opaque cloudy The examination can be performed on all materials containing a sufficient number of lymphocytes (approx. 50,000 cells) (fixed and Total protein < 25 g/l > 30 g/l 25 - 75 g/l unfixed cell material/tissue/smears/EDTA blood). Rivalta reaction Negative Positive Variable As a cytological or a histological examination can reliably determine the presence of a lymphocyte population, these preliminary Cell count < 1000/µl > 5000/µl 1000 - 7000/µl examinations are highly recommended. In addition, the results of these pre-examinations (in addition to the clinical picture) should be 260 261 2019/20 Sex Determination in birds 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics LABORATORY FOR CLINICAL DIAGNOSTICS 19 Sex Determination in Birds 20 Hereditary Diseases/Phenotype/ The method we use to determine the gender is based on the principle of polymerase Breed Characteristics chain reaction (PCR). It allows for quick and reliable determination of the sex of the bird using small quantities of genome-containing samples. The test is based on the repli- cation of two highly conserved target genes, which makes it possible to examine many different species. 20.1 Heredities The method carried out by us provides double safety: During PCR, one probe specifi- cally binds to the “female” sequence, while the other one binds to the “male” sequence, Autosomal recessive inheritance if these are present. This way, there will be one sex confirmed and the other sex exclu- A recessive allele is overwritten by its dominant counterpart. Therefore, autosomal ded. recessive inherited diseases only affect animals which received two mutated alleles. With 50% probability carrier animals inherit the mutated allele to their offspring. Bree- ding two carriers results in 25% of the offspring being affected by the disease. In Which sample material is appropriate? healthy populations, carrier animals enlarge the gene pool and should not be excluded from breeding categorically. Never the less, breeding of carriers with other carriers or Sex determination can be done by using either blood or quills. One to three drops even affected animals should strictly be avoided. of whole blood (preferably EB) are sufficient. They can be collected in suitable micro capillary tubes or applied dropwise to a filter card. Filter cards/blood cards should be Autosomal dominant inheritance completely dry before shipment. A dominant allele overwrites its recessive counterpart. Therefore, autosomal dominant Alternatively, 2 – 3 quills from freshly plucked feathers are required. Lost feathers and inherited diseases affect animals which received one mutated allele. Either father or down feathers are not suitable for the test. mother of the affected animal have to carry the mutated allele and are affected by the To ensure a correct analysis, the sample must not be contaminated with foreign DNA. disease themselves. For this purpose, please wear gloves when sampling or wash your hands after each sample you take. Please pack feathers separately for each bird. For “dry” feathers, it Autosomal dominant inheritance with incomplete penetrance is sufficient to use an envelope or a small paper bag, “wet” feathers can, for example, Heterozygous animals can also show symptoms of the disease but with varying be packed in blood or urine tubes or in standard freezer bags. Additionally, we offer expression. so-called SampleKits for submitting feather samples or blood cards. You can request them free of charge. We recommend marking the samples with the ring or chip number X-chromosomal or gonosomal recessive inheritance of the bird. For gonosomal coupled traits, an animal derives an X- or Y-chromosome from the father and one of two X-chromosomes from its mother. The mutated allele resides on the X-chromosome only. Which bird species can be tested? We have performed sex determination tests for several years and thus have already tested many different bird species. Only after we tested a female and a male bird of a 20.2 Dog species, we give clearance for routine diagnostics. In some species, PCR differentiation is not possible. 20.2.1 Hereditary Diseases We will be pleased to provide you with information on which bird species we test. It is essential to specify the exact bird species when submitting the samples. Acatalasemia Material EB 1 ml, buccal swab Method Sequencing Breed Beagle Inheritance Autosomal recessive Duration 1 – 2 weeks 262 263 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Note Acatalasemia is characterised by the deficiency of the enzyme Note The clinical signs of ARDS in Dalmatians are progressive tachypnea, catalase which is important for cellular defence in oxidative stress. dyspnea, respiratory distress, pulmonary lesions and some affected Affected dogs suffer from ulcers and tissue death in the oral cavity. puppies also showed renal aplasia and hydrocephalus. The typical age of onset is about 5 – 10 months. The puppies usually have to be Achromatopsia/Day Blindness (ACHM) euthanised 1 – 6 weeks after the appearance of first symptoms. The described mutation is so far not found in other breeds than dalmati- Material EB 1 ml, buccal swab an. Method Sequencing Breed German Shepherd, Labradoodle, Labrador Retriever Inheritance Autosomal recessive Alaskan Husky Encephalopathy (AHE) Duration 1 – 2 weeks Material EB 1 ml Note Achromatopsia ACHM is a disease in which the cone cells of the Method Sequencing retina, which are responsible for colour vision, are not developed Breed Husky properly. Inheritance Autosomal recessive Initial symptoms are noticeable in affected dogs from 8 – 10 weeks Duration 1 – 2 weeks of age. In low light conditions their visual function is comparable to Note A fatal brain disease known as Alaskan Husky encephalopathy normal dogs. (AHE) is found in Husky breeds. For AHE affected dogs, clinical fin- dings consist of multifocal central nervous system deficits including Acral Mutilation Syndrome (AMS) seizures, altered mentation, dysphagia, absent menace response, central blindness, hypermetria, proprioceptive positioning deficits, Material EB 1 ml, buccal swab facial hypoalgesia, ataxia and tetraparesis. Method TaqMan SNP assay Breed English Cocker Spaniel, English Pointer, English Springer Spaniel, French Spaniel, German Shorthaired Pointer Alaskan Malamute Polyneuropathy (AMPN) Inheritance Autosomal recessive Material EB 1 ml, buccal swab Duration 3 – 5 days Method TaqMan SNP-assay Note The Acral mutilation syndrome (AMS) is characterised by a senso- Breed Alaskan Malamute ry neuropathy of the peripheral parts of the body, such as limbs, Inheritance Autosomal recessive fingers or toes. Affected puppies show an insensitivity to pain in their Duration 3 – 5 days distal extremities. However, the disease often emerges not until the Note AMPN is a neurological disease which leads to progressive muscle age of about 4 months, when puppies begin to lick, bite or even self- weakness and exercise intolerance as well as signs of paralysis and mutilate their distal extremities. The proprioception, motor abilities respiratory problems at a later stage. and spinal reflexes remain intact. Moreover, the pain perception becomes progressively normal above the knees and is not altered in Alexander Disease (AxD) the rest of the body. Some affected dogs show only insensitivity to pain but no self-mutilation, making diagnosis extremely difficult. Material EB 1 ml, buccal swab Method Sequencing Breed Labrador Retriever Acute Respiratory Distress Syndrome (ARDS) Inheritance Autosomal dominant Material EB 1 ml, buccal swab Duration 1 – 2 weeks Method Sequencing Note The homozygous affected Labrador Retriever developed progres- Breed Dalmatian sively worsening tetraparesis with a spastic position of the thoracic Inheritance Autosomal recessive limbs and flattened chest. Later on, myoclonic jerks at the head and Duration 1 – 2 weeks the cervical region, absent patellar reflexes, weakness on the four limbs and mild generalised muscle atrophy became obvious. 264 265 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Amelogenesis Imperfecta (AI) C3 Deficiency (C3) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method Sequencing Breed Italian Greyhound, Samoyed Breed Brittany Spaniel Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 1 – 2 weeks Note AI is an inherited condition of tooth enamel malformation. Affected Note The third component (C3) of the complement system plays an animals have slim, pointed teeth with thin brown tooth enamel. Des- important role in the immune response to microbes like bacteria, pite this condition the teeth do not seem to be more susceptible for fungi or parasites. The causative mutation prohibits the formation of dental cavities. functional C3 units and the chain of defence mechanisms is broken. Affected dogs exhibit higher susceptibility to bacterial infections, e.g. Bardet-Biedl Syndrome (BBS) glomerulonephritis. Material EB 1 ml, buccal swab Method Sequencing Canine Leucocyte Adhesion Deficiency (CLAD) Breed Puli Material EB 1 ml, buccal swab Inheritance Autosomal recessive Method TaqMan SNP Assay Duration 1 – 2 weeks Breed Irish Red and White Setter, Irish Setter Note BBS is characterised by retinopathy, obesity and infertility. These ver- Inheritance Autosomal recessive satile symptoms occur in variable forms in affected dogs. First signs Duration 3 – 5 days of the disease can be detected already in the puppies. Note CLAD is a fatal immunodeficiency disease. Because their healing capacities are impaired, the affected dogs show severe infections Brachyuria (stumpy tail) (omphalophlebitis, skin infections, osteomyelitis and gingivitis). At the age of 8 – 12 weeks, a swelling of the jawbones and joint inflam- Material EB 1 ml, buccal swab mations could be observed leading to the CLAD typical unsteady Method TaqMan SNP assay gait. Breed Australian Shepherd, Australian Stumpy Tail Cattle Dog, Austrian Pin- scher, Bourbonnais Pointing Dog, Bouvier des Ardennes, Brazilian Terrier, Brittany Spaniel, Cardigan Welsh Corgi, Croatian Sheepdog Canine Multi-focal Retinopathy (CMR1/2/3) (Hrvatski Ovcar), Danish-Swedish Farmdog, Jack Russell Terrier, Material EB 1 ml, buccal swab Karelian Bear Dog, Miniature Australian Shepherd, Mudi, Pembroke Method TaqMan SNP assay (Coton de Tulear); sequencing Welsh Corgi, Polish Lowland Sheepdog (PON), Pyrenean Sheep- Breed American Bulldog, Australian Shepherd, Boerboel, Bullmastiff, Coton dog, Savoy Sheepdog, Schipperke, Spanish Water Dog, Swedish de Tuléar, Dogo Canario, Dogue De Bordeaux, English Bulldog, Vallhund Finnish Lapphund, French Bulldog, Italian Corso Dog, Lapponian Inheritance Autosomal dominant Herder, Mastiff, Miniature Australian Shepherd, Pyrenean Mountain Duration 3 – 5 days Dog, Swedish Lapphund Note Particularly the tail length bestows the characteristic look of most Inheritance Autosomal recessive dog breeds. To dock the tail is forbidden in most countries. A gene- Duration 3 – 5 days (Coton de Tulear); tic analysis allows the confirmation of a natural origin of a short tail. 1 – 2 weeks Note CMR is an inherited disease which causes lesions of the retina. First symptoms can be found at the age of about four months. In some cases, lesions disappear for some time and emerge again later on. Blindness or visual impairment was not found in affected dogs yet. 266 267 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Canine Multiple System Degeneration (CMSD) Note Chondrodysplasia is a genetically determined skeletal dysplasia which leads to defects in tubular bones and dwarfism. In addition to Material EB 1 ml, buccal swab shortened limbs, clinical symptoms comprise of a large skull, spine Method Sequencing changes and deformations of the legs. Breed Chinese Crested Dog, Kerry Blue Terrier Inheritance Autosomal recessive Duration 1 – 2 weeks Chondrodysplasia and -dystrophia (IVDD risk) Note CMSD is a fatal, hereditary movement disorder. Affected dogs Material EB 1 ml, buccal swab develop cerebellar ataxia at 3 – 6 months of age. First clinical sign is Method FLP typically an intention tremor of the head, followed by goose-stepping Breed All gait in the thoracic limbs. Inheritance CDPA autosomal dominant; CDDY semi dominant (leg length) or Most dogs were euthanised at 1 – 2 years of age. dominant (IVDD risk) Duration 1 – 2 weeks Centronuclear Myopathy (CNM) Note In many dog breeds, chondrodystrophy (CDDY) and/or chondro dysplasia (CDPA) cause a short leg phenotype. However, CDDY is Material EB 1 ml, buccal swab associated with a high risk of premature degeneration of the inverte- Method Sequencing (1); FLP (2) bral discs (Hansen’s type I intervertebral disc disease, IVDD). Breed German Hunting Terrier, Great Dane (1), Labrador Retriever (2) We offer the combined test for CDPA and CDDY in order to enable Inheritance Autosomal recessive the prevention of CDDY and to prefer CDPA, so that the short leg Duration 1 – 2 weeks phenotype could be maintained (by CDPA) while minimising the risk Note CNM as a disease causes dysplasia of the dogs muscles. There- for IVDD. fore, affected dogs lack tendon reflexes and gain less weight than puppies of the same age (4 weeks). Obvious symptoms of HMLR Cleft Lip/Palate and Syndactyly (CLPS) first occur at weeks 12 to 20 and include general amyosthenia, aber- rant bearing, awkward gait and trouble with ingestion. Material EB 1 ml, buccal swab Method TaqMan SNP assay Breed Nova Scotia Duck Tolling Retriever Cerebral Dysfunction (CDFS) Inheritance Autosomal recessive Material EB 1 ml, buccal swab Duration 3 – 5 days Method TaqMan SNP assay Note Cleft lip/palate and syndactyly (CLPS) describes a hereditary Breed Stabijhoun disease in Nova Scotia Duck Tolling Retrievers. Inheritance Autosomal recessive Early on, affected puppies develop a cleft palate, cleft lips, as well Duration 3 – 5 days as the pathological growth along the middle toes (syndactyly). Note CDFS in the Stabijhoun breed is an inherited disease of the brain. Clinically affected animals exhibit a large variety of neuronal Collie-Eye-Anomaly* (CEA) Optigen symptoms such as depressed behaviour, walking circles, obsessive sniffing and running backwards. Material EB 1 ml, buccal swab Method Optigen or partner laboratory Breed Australian Shepherd, Bearded Collie, Border Collie, Boykin Spaniel, Chondrodysplasia Collie (rough/smooth), Hokkaido, Lancashire Heeler, Long-Haired Material EB 1 ml, buccal swab Whippet (Silken Windsprite), Miniature Australian Shepherd, Nova Method Sequencing Scotia Duck Tolling Retriever, Shetland Sheepdog (Sheltie), Silken Breed Chinook, Karelian Bear Dog, Norwegian Elkhound Windhound Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 4 – 6 weeks 268 269 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Note CEA is a hereditary canine ocular disorder in which the pattern of Note Lamellar ichthyosis so far has only been observed in Great Danes. chorioretinal and scleral development is variously disturbed. The The skin becomes dry and loses its elasticity, whereby the animal major change in affected dogs is hypoplasia (underdevelopment) of exhibits a wrinkled look (especially the head is affected). In addition, the choroid. A Coloboma, or hole, may form in or near the optic disc puppies sometimes develop a strong swelling of the eyelids. The as part of the CEA extended phenotype. The degree of these ab changes in skin-properties causing the folds also enhances the risk normalities varies, ranging from mild disease to complete blindness. of secondary infections. The abnormality can be diagnosed at a very young age and is not progressive. Congenital Myasthenic Syndrome (CMS) Material EB 1 ml, buccal swab Cone Degeneration* (CD) Optigen Method Sequencing Material EB 1 ml, buccal swab Breed Golden Retriever, Jack Russell Terrier, Labrador Retriever, Old Danish Method Partner laboratory (Optigen) Pointing Dog, Parson Russell Terrier Breed Alaskan Malamute, Australian Shepherd, German Shorthaired Inheritance Autosomal recessive Pointer, Miniature Australian Shepherd Duration 1 – 2 weeks Inheritance Autosomal recessive Note Symptoms of the disease are most of all a generalised skeletal Duration 3 – 4 weeks muscle weakness, which worsens especially after exercise, stress Note Cone degeneration (CD) is an inherited disease of the retina. Cone or excitement. Signs can already be seen at just two weeks of age. cells start to degenerate in the puppy-stage already, leading to a Mobility of all limbs is reduced, and the ability to bear the own body distinct form of dayblindness. Affected puppies avoid bright light, weight diminishes with time. In all limbs, spinal reflexes are reduced. as those conditions can cause pain for the dog. Vision in dim light conditions or at night stays normal. First symptoms can be Congenital Stationary Night Blindness (CSNB) recognised at the age of about 8 – 12 weeks. Material EB 1 ml, buccal swab Method TaqMan SNP assay Congenital Hypothyroidism with Goiter (CHG) Breed Briard Material EB 1 ml, buccal swab Inheritance Autosomal recessive Method Sequencing Duration 3 – 5 days Breed Fox Terrier, French Bulldog, Rat Terrier, Spanish Water Dog, Note The Briard suffers from a retinal disorder characterised by congeni- Tenterfield Terrier, Toy Fox Terrier tal night blindness with various degrees of visual impairment under Inheritance Autosomal recessive photo topic illumination. The vision of affected dogs can range from Duration 1 – 2 weeks normal day vision to profound day blindness. The disease generally Note Affected dogs have a very short lifespan and normally die as appears first in early whelpage, at about six months and results from puppies. Daily medication with thyroid hormones can prolong the a mutation in the RPE65 gene. lifespan, still the dogs will suffer from dwarfism and goiter. Copper Storage Disease – Copper Toxicosis (CT) Congenital Ichthyosis Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method FLP Method Sequencing Breed Bedlington Terrier Breed Great Dane Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 1 – 2 weeks 270 271 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Note Copper toxicosis in Bedlington Terriers is based on a disturbance of Note DWLM is caused by a malformation of the cerebellum. Symptoms the copper metabolism, resulting in an accumulation of copper in include various forms of ataxia and commence at the age of 5 – 6 the liver and other organs. weeks. Affected dogs exhibit wobbly gait, loss of balance or periodic The genetic test identifies a mutation, which represents a major risk reoccurring collapses. Epileptic seizures were described in some factor for the development of the disease. cases as well. Craniomandibular Osteopathy (CMO) Degenerative Myelopathy (DM exon 1 and 2) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay Method TaqMan SNP assay Breed Cairn Terrier, Scottish Terrier, West Highland White Terrier Breed All (exon 2) Inheritance Autosomal dominant with variable penetrance Bernese Mountain Dog (1+2) Duration 3 – 5 days Inheritance Autosomal recessive with variable penetrance; The mutation in the Note Craniomandibular osteopathy (CMO) is a painful proliferative bone SOD1-gene which represents a major risk factor for the development disease affecting dogs of both sexes less than 1 year of age. of the disease is identified in the genetic test. Clinically, the disease is characterised by intermittent episodes of Duration 3 – 5 days concurrent fever and painful mandibular swelling. Note DM is a fatal, slowly progressing neuro degenerative disease with a late onset (age 8 years or older). The initial clinical sign is cha- Cystinuria racterised by spastic and general proprioceptive ataxia of the hind limbs. As the disease progresses, the frequently observed asym- Material EB 1 ml, buccal swab metric weakness ascends to affect the thoracic limbs, resulting in Method TaqMan SNP assay (1); sequencing (2) paraplegia. Hyporeflexia of the myotactic and withdrawal reflexes Breed Continental Bulldog, English Bulldog, French Bulldog, Landseer, occur. Mastiff, Newfoundland, Olde English Bulldog (1); Labrador Retriever, DM exon 2: Laboklin owns the exclusive license to perform this Australian Cattle Dog, Miniature Pinscher (2) genetic test. Inheritance Autosomal recessive; autosomal dominant in Australian Cattle Dog, Miniature Pinscher Duration 3 – 5 days (Continental Bulldog, English Bulldog, French Bulldog, Digital Hyperkeratosis (DH/HFH) Landseer, Mastiff, Newfoundland, Olde English Bulldog); Material EB 1 ml, buccal swab 1 – 2 weeks (Labrador Retriever, Australian Cattle Dog, Miniature Method TaqMan SNP assay Pinscher) Breed Dogue De Bordeaux, Irish Terrier, Kromfohrländer Note Cystinuria is an inherited disorder caused by a defective transport of Inheritance Autosomal recessive the amino acid cystine in the kidney tubules which leads to formation Duration 3 – 5 days of cystine calculi (stones). Note This disease is also called “corny feet” as the main symptom is thickening and hardening of the footpads. First symptoms appear Dandy-Walker-like Malformation (DWLM) at an age of 10 weeks to 12 months (Dogue de Bordeaux) and at an age of 4 – 9 months (Irish Terrier and Kromfohrländers). Affected Material EB 1 ml, buccal swab dogs suffer from painful cracks and crevices sometimes leading to Method TaqMan SNP assay secondary infections. Additionally, nail growth is often accelerated. Breed Eurasien Inheritance Autosomal recessive Duration 3 – 5 days 272 273 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Dry Eye Curly Coat Syndrome (CCS) Note The keratin defect leads to superficial, mild, plantar epidermolytic hyperkeratosis with epidermal fragility. Affected dogs show clinical Material EB 1 ml, buccal swab signs from birth through adulthood. Method TaqMan SNP assay Breed Cavalier King Charles Spaniel Inheritance Autosomal recessive Episodic Falling (EF) Duration 3 – 5 days Material EB 1 ml, buccal swab Note Affected dog puppies suffer from keratoconjunctivitis sicca from Method TaqMan SNP assay eyelid opening, abnormal (rough/curly) coat and are usually smaller Breed Cavalier King Charles Spaniel than littermates. Footpads are hyperkeratinised from young adult- Inheritance Autosomal recessive hood including nail growth abnormalities and intermittent sloughing, Duration Approx. 1 week pain and lameness. Note Episodic falling is a neurological disorder. Episodes are triggered by exercise, stress or excitement and are characterised by stiffness Dystrophic Epidermolysis Bullosa (DEB) throughout thoracic and pelvic limbs resulting in a characteristic ‘deer-stalking’ position and/or collaps. Material EB 1 ml, buccal swab Laboklin owns the exclusive license to perform this genetic test. Method Sequencing Breed Central Asian Shepherd Dog Inheritance Autosomal recessive Exercise Induced Collapse (EIC) Duration 1 – 2 weeks Material EB 1 ml, buccal swab Note Epidermolysis bullosa is characterised by blisters in the dermal- Method TaqMan SNP assay epidermal junction. The disease onset varies between birth and early Breed Boykin Spaniel, Chesapeake Bay Retriever, Clumber Spaniel, Curly infancy. Affected puppies show skin lesions, blisters and ulcers on Coated Retriever, German Wirehaired Pointer, Labrador Retriever, feet, ears, muzzle and oral mucosa. Affected dogs were euthanised Old English Sheepdog, Welsh Corgi Pembroke due to an unfavourable prognosis. Inheritance Autosomal recessive Duration 3 – 5 days Ectodermal Dysplasia/Skin Fragility Syndrome (ED/SFS) Note The syndrome of Exercise induced collapse (EIC) is manifested by muscle weakness, incoordination and life-threatening collapse Material EB 1 ml, buccal swab after intense exercise. Five to fifteen minutes of strenuous exercise Method Sequencing causes dogs suffering from this condition to develop a „wobbly“ Breed Chesapeake Bay Retriever gait, which soon progresses to nonpainful, flaccid paraparesis and a Inheritance Autosomal recessive loss of control of the rear limbs. Laboklin owns the exclusive license Duration 1 – 2 weeks to perform this genetic test. Note At birth, the skin of affected dogs is abnormally pale and translu- cent on ears, feet, nose and mouth area. Spontaneous sloughing Factor VII Deficiency of the nose and footpad epithelium and bleeding of ear tips occur if traumatised. Lips and facial skin layers also slough when rubbed or Material EB 1 ml, buccal swab licked by mother. Affected dogs have to be euthanised. Method TaqMan SNP assay Breed Airedale Terrier, Alaskan Klee Kai, Beagle, Deerhound, Finnish Hound, Giant Schnauzer, Welsh Springer Spaniel Epidermolytic Hyperkeratosis (EHK) Inheritance Autosomal recessive Material EB 1 ml, buccal swab Duration 3 – 5 days Method Sequencing Note Affected dogs show a mild to moderate bleeding tendency but can Breed Norfolk Terrier also remain asymptomatic. Inheritance Autosomal recessive Duration 1 – 2 weeks 274 275 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Familial Nephropathy (FN) Fucosidosis Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Partner laboratory* (English Cocker Spaniel, Welsh Springer Spaniel) Method Sequencing Sequencing (English Springer Spaniel, Samoyed) Breed English Springer Spaniel Breed English Cocker Spaniel, English Springer Spaniel, Samoyed, Welsh Inheritance Autosomal recessive Springer Spaniel Duration 1 – 2 weeks Inheritance Autosomal recessive (English Cocker Spaniel, English Springer Note Fucosidosis is a lysosomal storage disorder causing deposits Spaniel, Welsh Springer Spaniel) in cerebral and neural tissue. Affected animals show a disturbed X-chromosomal recessive in Samoyed coordination of movements, behavioural abnormalities, blindness, Duration 1 – 2 weeks deafness and problems in deglutition. The disease occurs between Note Dogs with FN typically develop chronic renal failure between 6 the age of 18 months and 4 years with a progressive course and months and 2 years of age, with eventual and sometimes rapid finally lethal outgoing. destruction of both kidneys. Gallbladder Mucoceles Fanconi Syndrome Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method Sequencing Breed American Cocker Spaniel, Cairn Terrier, English Cocker Spaniel, Breed Basenji Pomeranian, Shetland Sheepdog (Sheltie) Inheritance Unknown Inheritance Autosomal dominant with incomplete penetrance Duration 1 – 2 weeks Duration 1 – 2 weeks Note Fanconi syndrome is an inherited disease in the Basenji. Due to Note Undetected gallbladder mucoceles can lead to cholecystitis and renal dysfunction, electrolytes and nutritive substances cannot be thus increase the risk of a rupture of the gallbladder. Clinical signs reabsorbed from the urine. Excessive uptake of water and urinating occur in older dogs showing vomiting, anorexia, lethargy, icterus and are typical symptoms. Without treatment, affected dogs exhibit abdominal pain. amyosthenia, acidosis and general weakness that leads to death ultimately. In Basenji dogs, onset of Fanconi syndrome is found at Glanzmann Thrombasthenia (GT) the age of four to eight years. Material EB 1 ml, buccal swab Method Sequencing Finnish Hound Ataxia (FHA) Breed Pyrenean Mountain Dog Material EB 1 ml, buccal swab Inheritance Autosomal recessive Method TaqMan SNP assay Duration 1 – 2 weeks Breed Finnish Hound, Norrbottenspitz Note GT is a bleeding disorder that occurs in two different types. They Inheritance Autosomal recessive differ in the amount of specific glycoproteins (aIIbb3) embedded Duration 3 – 5 days in the cell membrane of platelets (thrombocytes), which are im- Note Affected dogs will show first indications of cerebellar neurodege- portant part of the coagulation system. In the severe GT of type I neration at the age of 4 – 12 weeks. First clinical signs are loss of this amount of glycoprotein is less than 5% of the normal value. A balance, minor incoordination of gait and intention tremor while later mutation in the αIIb gene disrupts the production of one main com- symptoms can be a progressive incoordination or a complete loss of ponent of these glycoproteins. Symptomatically, GT shows in form of mobility. bleeding diathesis and continuous gingival bleeding after shedding of deciduous teeth. Continuous epistaxis can also be an indication for this disorder. 276 277 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Glaucoma and Goniodysgenesis (GG) Glycogen Storage Disease GSD II (Pompe Disease) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay Method Sequencing Breed Border Collie Breed Finnish Lapphund, Lapponian Herder, Swedish Lapphund Inheritance Presumably autosomal recessive (still in research). Inheritance Autosomal recessive Duration 3 – 5 days Duration 1 – 2 weeks Note Goniodysgenesis is a development abnormality of the anterior Note Affected dogs suffer from vomiting, progressive muscle weakness chamber of the eye, characterised by narrowing or closure of and loss of condition. Heart disease leads to death at about 1.5 intraocular channels of the iridocorneal angle. The increased ocular years of age. pressure damages the retinal ganglion and can therefore result in primary glaucoma and blindness. The development of a glaucoma Glycogen Storage Disease GSD IIIa seems to be influenced by a combination of genetic, environmental and/or random factors. Moreover, the OLFML3 variant could not be Material EB 1 ml, buccal swab associated with mild goniodysgenesis indicating that the mild form Method Sequencing is caused by another genetic disposition. Breed Curly Coated Retriever Inheritance Autosomal recessive Duration 1 – 2 weeks Globoid Cell Leucodystrophy (Krabbe Disease) Note Affected puppies don´t exhibit symptoms during the first years. The Material EB 1 ml, buccal swab disease gets obvious when lethargy and episodical hypoglycaemia Method TaqMan SNP assay (Cairn Terrier, West Highland White Terrier) including collapses occur after some years. Sequencing (Irish Setter) Breed Cairn Terrier, Irish Setter, West Highland White Terrier Inheritance Autosomal recessive GM1-Gangliosidosis (GM1) Duration 3 – 5 days (Cairn Terrier, West Highland White Terrier) Material EB 1 ml, buccal swab 1 – 2 weeks (Irish Setter) Method Sequencing Note The globoid cell leucodystrophy manifests itself in affected dogs at Breed Husky, Portuguese Water Dog, Shiba the age of 1 – 3 months, beginning with ataxia and paresis of the Inheritance Autosomal recessive hind legs. During the progress of the disease, muscular atrophy and Duration 1 – 2 weeks neurological degeneration occur. Due to the lack of treatment possi- Note GM1 is a lysosomal storage disease that leads to neurological bilities, the affected animals are usually euthanised after 10 months disorders. Affected dogs suffer from paralysis of the extremities and at the latest. spasticity of the muscles. Glycogen Storage Disease GSD I GM2-Gangliosidosis (GM2) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method Sequencing Breed Maltese Breed Japanese Chin, Miniature Poodle, Shiba, Toy Poodle Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 1 – 2 weeks Note Glycogen storage disease type I (GSD I) is caused by a dysfunction Note GM2 gangliosidosis is a fatal, progressive neurodegenerative of the glucose metabolism, which leads to mass storage of glyco- lysosomal storage disease. First symptoms appear at the age of 9 gen in liver, muscle and nerve cells, resulting in gradual dysfunction to 12 months and typically include loss of vision, walking difficulties, (variable severity) of these organs. Affected puppies exhibit symp- loss of balance, head tremors and vomiting. The disease progresses toms like depression, bad nutritional state and slow growth. rapidly and dogs usually die between the age of 18 and 23 months. 278 279 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Grey Collie Syndrome (GCS) (Canine Cyclic Neutropenia) Note The enhanced tendency to haemorrhage is caused by impaired coagulation activity, recognisable by activated platelets, which are Material EB 1 ml, buccal swab not capable to present anionic phospholipids, particularly phos- Method Sequencing phatidylserine, and to distribute coagulation microparticles. Other Breed Collie (rough/smooth) coagulation parameters are normal, with the exception of a reduced Inheritance Autosomal recessive prothrombin consumption during clotting of blood. Duration 1 – 2 weeks Note The disorder is characterised by an abnormality of the stem cells in Hereditary Ataxia (HA) the bone marrow. Affected dogs are susceptible to severe recurring bacterial infections and they are also prone to bleeding episodes. Material EB 1 ml, buccal swab Method TaqMan SNP assay Breed Gordon Setter, Old English Sheepdog Hemophilia A (Factor VIII Deficiency) Inheritance Autosomal recessive Material EB 1 ml, buccal swab Duration 3 – 5 days Method TaqMan SNP assay (German Shepherd) Note Affected dogs will show first indications of cerebellar neurodegene- FLP (Havanese) ration at an age between six months and up to four years. Clinical Sequencing (Boxer, Old English Sheepdog) signs are loss of balance, minor incoordination of gait and intention Breed Boxer, German Shepherd, Havanese, Old English Sheepdog tremor, which later progress to severe gait disturbances. Inheritance X-chromosomal recessive Duration 1 – 2 weeks Hereditary Cataract (HSF4) Note Affected dogs present with haemorrhage that can vary from mild to severe depending on the degree of the disease. Material EB 1 ml, buccal swab Laboklin owns the exclusive license to perform this genetic test. Method Partner laboratory* (Boston Terrier, French Bulldog, Staffordshire Bull Terrier) Sequencing (Australian Shepherd, Miniature Australian Shepherd, Hemophilia B (Factor IX Deficiency) Wäller) Material EB 1 ml, buccal swab Breed Australian Shepherd, Miniature Australian Shepherd, Wäller Method TaqMan SNP assay (Rhodesian Ridgeback) Inheritance Autosomal recessive (Boston Terrier, French Bulldog, Staffordshire Sequencing (Lhasa Apso) Bull Terrier) Breed Hovawart, Lhasa Apso, Rhodesian Ridgeback Autosomal dominant with incomplete penetrance (Australian Inheritance X-chromosomal recessive Shepherd, Miniature Australian Shepherd, Wäller) Duration 3 – 5 days (Rhodesian Ridgeback) Duration 1 – 2 weeks 1 – 2 weeks (Hovawart, Lhasa Apo) Note The cataract is one of the most frequent causes for blindness in Note Affected dogs present with haemorrhage that can vary from mild to dogs. severe depending on the degree of the disease. Laboklin owns the exclusive license to perform this genetic test. Hereditary Deafness (PTPRQ) Material EB 1 ml, buccal swab Haemorrhagic Diathesis (Scott Syndrome) Method Sequencing Material EB 1 ml, buccal swab Breed Dobermann Method Sequencing Inheritance Autosomal recessive Breed German Shepherd Duration 1 – 2 weeks Inheritance Autosomal recessive Duration 1 – 2 weeks 280 281 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Note Affected pups are deaf by 3 weeks of age and show signs of Hypomyelination/Shaking Puppy Syndrome (SPS) vestibular disease like head tilt, circling and ataxia. In the inner ear, progressive cochlear degeneration with a loss of auditory sensory Material EB 1 ml, buccal swab cells has been described. Moreover, abnormal or absent otoconia Method Sequencing could be found in the vestibular system of some affected dogs Breed English Springer Spaniel, Weimaraner without sensory cell loss. Inheritance Autosomal recessive Duration 1 – 2 weeks Hereditary Nasal Parakeratosis (HNPK) Note The disease is caused by disruption of myelination in the spinal cord. Affected dogs show a generalised tremor at age of 12 to 14 Material EB 1 ml, buccal swab days. The severity of the tremor can vary between litter mates. The Method TaqMan SNP assay (Labrador Retriever) dogs can ambulate, but show a “hopping” gait in the hind limbs. Sequencing (Greyhound) The tremor is not present when the dogs are at rest or asleep and Breed Labrador Retriever, Greyhound diminishes over time by 3 to 4 months of age. Inheritance Autosomal recessive Duration 3 – 5 days (Labrador Retriever) Ichthyosis in Great Danes Ø see Congenital ichthyosis 1 – 2 weeks (Greyhound) Note Affected dogs develop first symptoms at the age of six months to Ichthyosis (American Bulldog) one year. The dorsal surface of the planum forms keratinous scales, which adhere to the nose. Material EB 1 ml, buccal swab Treatment can only be symptomatic. Method Sequencing Laboklin owns the exclusive license to perform this genetic test. Breed American Bulldog Inheritance Autosomal recessive Duration 1 – 2 weeks Hereditary Neuropathy (GHN) Note Ichthyosis is a genetic disease, causing dysfunction of the keratin Material EB 1 ml, buccal swab in the skin, which leads to the production of large, differently Method FLP pigmented skin scales. Additionally, the pigmentation of the skin can Breed Greyhound be altered. Dogs which are affected by this dermatosis develop first Inheritance Autosomal recessive symptoms soon after birth. Duration 1 – 2 weeks Note Symptoms include progressive amyosthenia, exercise intolerance, Ichthyosis* loss of reflexes and ataxia of all limbs. Material EB 1 ml, buccal swab Method Partner laboratory Hyperuricosuria (HUU/SLC) Breed Golden Retriever Material EB 1 ml, buccal swab Inheritance Autosomal recessive Method TagMan SNP assay Duration 1 – 2 weeks Breed All Note See Ichthyosis in American Bulldogs. Inheritance Autosomal recessive Duration 3 – 5 days Imerslund-Gräsbeck Syndrome (IGS) Note Hyperuricosuria is a metabolic disorder that leads to an increased excretion of uric acid instead of allantoin. Preventative, affected dogs Material EB 1 ml, buccal swab should get a low purine diet. Additionally, adequate hydration is vital. Method TaqMan SNP assay Breed Beagle, Border Collie Inheritance Autosomal recessive Duration 3 – 5 days 282 283 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Note The malabsorption of Vitamin B12 leads to neurological symptoms Juvenile Laryngeal Paralysis & Polyneuropathy (JLPP) and irreversible damage of the brain and nervous system. Material EB 1 ml, buccal swab Method TaqMan SNP assay Junctional Epidermolysis Bullosa (JEB) Breed Black Russian Terrier, Rottweiler Material EB 1 ml, buccal swab Inheritance Autosomal recessive Method TaqMan SNP assay Duration 3 – 5 days Breed German Shorthaired Pointer Note JLPP is a hereditary disease that leads to breathing difficulties when Inheritance Autosomal recessive; the genetic test detects a SNP mutation that is excited or physically stressed already at an age of around three inherited together with the responsible mutation. months. With progression of the disease, weakness and coordina- Duration 3 – 5 days tion problems in the hind limbs will develop, which eventually will Note JEB is a blistering disorder of the skin and mucous membranes in extend to the front limbs. Swallowing difficulties will make the puppy which tissue separation occurs within the lamina lucida of the base- prone to choking or pneumonia. This disease cannot be cured and ment membrane zone at the dermal-epidermal junction. Affected is fatal within a few months after first symptoms are detected. dogs show erosion and encrustation in the field of the footpads, knees, elbows, ankle joints, carpal bones, hips, lips and tongue. Juvenile Myoclonic Epilepsy (JME) Material EB 1 ml, buccal swab Juvenile Brain Disease (JBD) Method TaqMan SNP assay Material EB 1 ml, buccal swab Breed Rhodesian Ridgeback Method TaqMan SNP assay Inheritance Autosomal recessive Breed Jack Russell Terrier, Parson Russell Terrier Duration 3 – 5 days Inheritance Autosomal recessive Note JME in Rhodesian Ridgebacks is characterised by a particular Duration 3 – 5 days epilepsy phenotype of frequent myoclonic jerks with a mean onset Note The juvenile encephalopathy in Russell Terriers is a severe brain at 6 months of age. The dogs suffer from automatic and sudden disorder with an early onset at 6 – 12 weeks of age. The affected twitches, notably occuring in recumbent and relaxed situations. The puppies suffer from epileptic seizures. The disease progresses attacks arise daily in over 85% of cases. The disease progresses to rapidly causing irreversible brain damage leading to death. generalised tonic-clonic seizures in about 40% of affected dogs. Juvenile Epilepsy (JE) L-2-hydroxyglutaric Aciduria (L2HGA) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay Method TaqMan SNP assay Breed Lagotto Romagnolo Breed Staffordshire Bull Terrier Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 3 – 5 days Duration 3 – 5 days Note L-2-HGA produces a variety of neurological deficits, including Note Juvenile epilepsy occurs during week five to twelve of a puppys life. psychomotor retardation, seizures and ataxia. Symptoms are Typically, symptoms range from slight shivering and wobbly gait to „wobbly“ gait, tremors, muscle stiffness as a result of exercise or spastic paralysis. excitement, and altered behaviour. 284 285 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Lafora Disease (EB only) Leonberger Polyneuropathy 1 (LPN1) and 2 (LPN2) Material EB 0.5 – 1 ml, no buccal swabs! Material EB 1 ml, buccal swab Method Special: FLP Method FLP (LPN1) Breed Basset Hound, Beagle, Cardigan Welsh Corgi, Chihuahua, Dachs- Sequencing (LPN2) hund (Dackel), French Bulldog, Long-Haired Dachshund, Welsh Breed Leonberger Corgi Pembroke, Short-Haired Dachshund, Wire-Haired Dachshund Inheritance Autosomal recessive (LPN1) Inheritance Autosomal recessive Autosomal dominant with incomplete penetrance (LPN2) Duration 2 – 3 weeks Duration 1 – 2 weeks Note Lafora disease is an inherited glycogen metabolism disorder that Note LPN1 starts at the age of 2 – 4 years (early onset), LPN2 at the age causes progressive myoclonic epilepsy. The symptoms of this of 6 years. The symptoms of this polyneuropathy include wobbly gait disease are: poor vision/blindness, generalised tonic-clonic seizures, and paralysis especially from the backhand. Affected dogs are not myoclonic jerks (often induced by light, sound or sudden move- able to stand up in the final stage of the disease. Breathing noises, ments in the visual field), panic attacks, dementia, aggression as strange barking voice and difficulties in swallowing are other typical well as faecal and urinary incontinence as a result of loss of house symptoms. training. The average onset of first symptoms is reported at 7 years. Lethal Acrodermatitis (LAD) Lagotto Storage Disease (LSD) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay Method TaqMan SNP assay Breed Bull Terrier, Miniature Bull Terrier Breed Lagotto Romagnolo Inheritance Autosomal recessive Inheritance Autosomal recessive with incomplete penetrance Duration 3 – 5 days Duration 3 – 5 days Note Lethal acrodermatitis is characterised by skin lesions on the feet and Note Lagotto storage disease (LSD) is a storage disease with neurode- on the face, leading to secondary infections with Malassezia and generative symptoms, which leads to cerebellar damage in affected Candida. In addition, the lesions cause hyperkeratosis of the foot- animals. These are the cause of dysfunction in movement coordi- pads and deformation of the nails. Affected puppies also suffer from nation and balance. In some affected dogs, abnormal eye motion diarrhoea, bronchopneumonia and a failure to thrive. First symptoms (nystagmus) as well as behavioural changes like aggression and occur in the first weeks of life. Affected puppies typically die before restlessness can be detected. First symptoms appear at an age of they reach the age of two years. four months to four years. Leukocyte Adhesion Deficiency Type III (LAD3) Late Onset Ataxia (LOA) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method TaqMan SNP assay Breed German Shepherd Breed Jack Russell Terrier, Parson Russell Terrier Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 3 – 5 days Note LAD3 is an inherited immunodeficiency disease. It is caused by a Note Affected dogs will show first indications of the ataxia at the age of recessive mutation that impairs cell to cell adhesion for leucocytes. 6 – 12 months. First clinical signs are loss of balance and minor Thereby, granulocytes are unable to migrate to the site of the incoordination of gait while later symptoms can be a progressive infection. The animals are unable to form pus or neutrophilia. incoordination or a complete loss of mobility. Affected dogs develop severe infections early in life, due to a weak immune system. Those infections persist even when high doses of antibiotics are applied. 286 287 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Leukoencephalomyelopathy (LEMP) Note MTC is a hereditary disorder in the formation of thrombocytes. Two mutations have been identified in the ß1-tubulin gene, one of Material EB 1 ml, buccal swab which is recessive, while the other one is dominant. Indication for Method Sequencing (Great Dane, Rottweiler) a congenital MTC is a reduced number of platelets, with counts TaqMan SNP assay (Leonberger) ranging between 100.000 and 50.000 per µl or even below. Breed Great Dane, Leonberger, Rottweiler Moreover, circulating platelets are larger than normal. Heterozygous Inheritance Autosomal recessive with reduced penetrance carriers of the dominant mutation have counts intermediate to Duration 3 – 5 days (Leonberger) affected and normal dogs. Affected dogs do not have a bleeding 1 – 2 weeks (Great Dane, Rottweiler) diathesis, but as treatment with antibiotics or steroids is impropriate for the congenital MTC, this genetic test should be considered as a Note Leukoencephalomyelopathy (LEMP) is a juvenile-onset neurodege- method of differential diagnosis. nerative disorder in the white matter of the central nervous system (CNS). Typical symptoms of LEMP are coordination and movement disorders. Only a few months after the first symptoms, the affected Malignant Hyperthermia (MH) dogs will not be able to stand up or to walk any more. The age of Material EB 1 ml, buccal swab onset is about 1 – 3 years. Method Sequencing Breed All Lundehund Syndrome (LHS) Inheritance Autosomal dominant Duration 1 – 2 weeks Material EB 1 ml, buccal swab Method Sequencing Note Maligant hyperthermia (MH) is an inherited disorder of skeletal Breed Norwegian Lundehund muscle characterised by hypercarbia, rhabdomyolysis, generalised Inheritance Autosomal recessive skeletal muscle contracture, cardiac dysrhythmia and renal failure, Duration 1 – 2 weeks that develops on exposure to succinylcholine or volatile anaesthetic agents. If narcotics and muscle relaxants are administered further Note LHS describes a specific assembly of symptoms for the Norwegian on, damages of nerve, liver and kidney tissue will occur with fatal Lundehund that resemble common features of a protein losing ente- outcome. ropathy (PLE). Those symptoms include intestinal lymphangiectasia, gastrointestinal disturbance, inflammatory bowel disease and mal- absorption. Additionally, general weakness, vomiting and oedema May-Hegglin Anomaly (MHA) are commonly associated with this disease. Material EB 1 ml, buccal swab Method Sequencing Macrothrombocytopenia (MTC) Breed Pug Dog Inheritance Autosomal recessive Material EB 1 ml, buccal swab Duration 1 – 2 weeks Method Sequencing Breed American Cocker Spaniel, Bichon Frisé, Boxer, Cairn Terrier, Note In animals with MHA a persistent thrombocytopenia as well as Cavalier King Charles Spaniel, Chihuahua, English Cocker Spaniel, greatly enlarged and in morphology variably altered platelets are Havanese, Jack Russell Terrier, Labrador Retriever, Maltese, found in haematological diagnosis. Consequently, these animals Miniature Poodle, Norfolk Terrier, Parson Russell Terrier, Poodle, Shih have prolonged coagulation time. In addition, cytoplasmic inclusions Tzu, Toy Poodle can be detected in neutrophils. Consequently, these animals have Inheritance Variable prolonged coagulation time when bleeding. Duration 1 – 2 weeks 288 289 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS MDR1-gene Defect* (Ivermectin Hypersensibility) Mucopolysaccharidosis Type VII (MPS 7) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Partner laboratory Method Sequencing Breed Australian Shepherd, Border Collie, Collie (rough/smooth), Elo, Ger- Breed Brazilian Terrier, German Shepherd man Shepherd, Long-Haired Whippet (Silken Windsprite), McNab Inheritance Autosomal recessive Shepherd/Border Collie, Miniature Australian Shepherd, Old English Duration 1 – 2 weeks Sheepdog, Shetland Sheepdog (Sheltie), Silken Windhound, White Note Clinical signs are corneal clouding and severe skeletal deformities. Swiss Shepherd, Wäller Affected dogs are unable to ambulate at several weeks to months of Inheritance Autosomal recessive, hypersensibility is to be expected in carriers age. too, though. Duration 1 – 2 weeks Muscular Dystrophy (MD) Note Neurotoxic symptoms include mydriasis, tremors, ataxia and ano- rexia. These symptoms occur at doses that are 1/200th of the dose Material EB 1 ml, buccal swab required to cause toxicity in other dogs. Neurological manifestations Method TaqMan SNP Assay (Landseer) of ivermectin in susceptible dogs also include hypersalivation, Sequencing (Cavalier King Charles Spaniel, Golden Retriever, blindness, coma, respiratory compromise and death. Dogs with Norfolk Terrier) the MDR1 gene defect suffer from multiple intolerance of drugs. An Breed Cavalier King Charles Spaniel, Golden Retriever, Landseer, Norfolk interaction with the multi-drug-resistence-transporter (MDR1) has Terrier been proven for more than 100 drugs. Inheritance X-chromosomal recessive (Cavalier King Charles Spaniel, Golden Retriever, Norfolk Terrier) Autosomal recessive (Landseer) Microphthalmia (RBP4) Duration 3 – 5 days (Landseer) Material EB 1 ml, buccal swab 1 – 2 weeks (Cavalier King Charles Spaniel, Golden Retriever, Method TaqMan SNP assay Norfolk Terrier) Breed Irish Soft Coated Wheaten Terrier Note Affected dogs show raised creatine kinase levels, muscle atrophy Inheritance Autosomal recessive with maternal influence with contractures, fibrosis and cardiomyopathy. Generally, first Duration 3 – 5 days symptoms appear at the age of three to six months. Affected dogs Note A genetic variant that causes a prenatal vitamin A deficiency can be usually die between the ages of 4 and 24 months. responsible for the onset of the disease. Affected puppies only develop symptoms of microphthalmia, if their Musladin-Lueke Syndrome (MLS) dam is affected by this disease, too. The disrupted transfer of vita- min A begins in the hepatic store of the mother and continues at the Material EB 1 ml, buccal swab placenta. Method TaqMan SNP assay Breed Beagle Inheritance Autosomal recessive Mucopolysaccharidosis Type IIIa (MPS3a) Duration 3 – 5 days Material EB 1 ml, buccal swab Note MLS manifests with extensive fibrosis of the skin and joints. By one Method Sequencing year of age the disease stabilises whereby arthrosis, stiffness, short Breed New Zealand Huntaway, Wire-Haired Dachshund outer digits as well as a typical flat head shape with higher ear set Inheritance Autosomal recessive and slanted eyes are characteristic for this disease. Duration 1 – 2 weeks Note Affected animals suffer from severe degeneration of the central nervous system. Symptoms start around 18 months of age and are characterised by neurologic deterioration including ataxia and often lead to death. 290 291 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Myostatin Mutation („Bully“-gene) Inheritance Autosomal recessive with variable penetrance; The genetic test identifies a mutation, which represents a major risk factor for the Material EB 1 ml, buccal swab development of the disease. Method Sequencing Duration 1 – 2 weeks Breed Whippet Inheritance Autosomal recessive Note This auto-immune disease appears to have no infectious cause, Duration 1 – 2 weeks but results from a genome driven immune-response, which attacks cells of the central nervous system. First symptoms usually appear Note A mutation in the Myostatin gene has been found which correlates to at the age of six months to three years and include disorientation, higher performance in dog-racing. Dogs that possess one of these instability and seizures. The genetic test identifies a mutation which “Bully” alleles predominantly appeared in the top-racing-class, while represents a major risk factor that could be associated with PDE dogs with two “Bully” alleles are extremely muscular, but slower (Pug Dog Encephalitis). when running. Nemalin Myopathy (NM) Myotonia Congenita Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method TaqMan SNP assay (Miniature Schnauzer) Breed American Bulldog Sequencing (Australian Cattle Dog, Labrador Retriever) Inheritance Autosomal recessive Breed Australian Cattle Dog, Labrador Retriever, Miniature Schnauzer Duration 1 – 2 weeks Inheritance Autosomal recessive Duration 3 – 5 days (Miniature Schnauzer) Note Affected dogs exhibit muscular disorders such as muscle weakness, 1 – 2 weeks (Australian Cattle Dog, Labrador Retriever) hypotonia, hypoventilation and swallowing dysfunction. Note This inherited pathogenic condition affects skeletal muscle ion chan- nels. Clinical signs are a stiff, lanky gait and difficulties in swallowing Neonatal Cortical Cerebellar Abiotrophy (NCCD) as well as excess salivation. All affected Miniature Schnauzers exhi- Material EB 1 ml, buccal swab bit an abnormal set of teeth and overbite, sometimes also abnormal Method FLP (Beagle) barking. Sequencing (Magyar Viszla) Breed Beagle, Magyar Vizsla Narcolepsy Inheritance Autosomal recessive Duration 1 – 2 weeks Material EB 1 ml, buccal swab Method Sequencing (Doberman Pinscher, Dachshund) Note Affected dogs exhibit symptoms soon after birth or in very early age. TaqMan SNP assay (Labrador Retriever) These include tremor, ataxia and spastic paralysis. Breed Dachshund, Dobermann, Labrador Retriever Inheritance Autosomal recessive Neonatal Encephalopathy with Seizures (NEWS) Duration 1 – 2 weeks (Doberman Pinscher, Dachshund) Material EB 1 ml, buccal swab 3 – 5 days (Labrador Retriever) Method TaqMan SNP assay Note The disease is characterised by daytime sleepiness, cataplexy and Breed Poodle striking transitions from wakefulness into REM sleep. Inheritance Autosomal recessive Duration 3 – 5 days Necrotizing Meningoencephalitis (NME/PDE) Note NEWS is a malformation of the cerebellum. Affected puppies are Material EB 1 ml, buccal swab relatively small and weak and they often die within the first week of Method Sequencing their lives. Those surviving this phase develop ataxia, tremor and Breed Pug Dog seizures. None of them survived to the eighth week of life. 292 293 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Neuroaxonal Dystrophy (NAD) Neuronal Ceroid Lipofuscinosis* (NCL) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay (Spanish Waterdog) Method Partner laboratory Sequencing (Papillon, Rottweiler) Breed American Staffordshire Terrier Breed Papillon, Rottweiler, Spanish Water Dog Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 3 – 5 days (Spanish Waterdog) Note See Neuronal ceroid lipofuscinosis (NCL) above. 1 – 2 weeks (Papillon, Rottweiler) Note NAD is characterised by a histologically distinct neurodegenerative Obesity pathology of the central and/or peripheral nervous system. As with most neurological disorders, symptoms may vary. The onset of the Material EB 1 ml, buccal swab disease takes place in early development. Homozygous puppies Method FLP die at birth due to respiratory failure and exhibit swollen axons and Breed Flat Coated Retriever, Labrador Retriever spheroids throughout the nervous system. Inheritance Yet unknown Duration About one week Neuronal Ceroid Lipofuscinosis (NCL) Note In the Labrador retriever breed and the closely related Flat coated retriever, a POMC (pro-opiomelanocortin) mutation has been found Material EB 1 ml, buccal swab to disrupt the production of the neuroactive peptides ß-MSH and Method Sequencing (Australian Cattle Dog, Australian Shepherd, Chinese ß-endorphin, both implicated in energy homeostasis. Furthermore, Crested Dog, Chihuahua, Dachshund, Golden Retriever, Saluki) the mutation is associated with greater weight, adiposity and food TaqMan SNP assay (American Bulldog, Border Collie, English Setter, motivation in affected dogs. Therefore, the mutation is especially Gordon Setter, Tibetan Terrier) common in assistance dogs. Breed American Bulldog, Australian Cattle Dog, Australian Shepherd, Border Collie, Chihuahua, Chinese Crested Dog, Dachshund (Dackel), English Setter, Golden Retriever, Gordon Setter, Long- Osteogenesis Imperfecta (Brittle Bone Disease) Haired Dachshund, Miniature Australian Shepherd, Saluki, Short- Material EB 1 ml, buccal swab Haired Dachshund, Tibetan Terrier, Wire-Haired Dachshund Method TaqMan SNP assay (Shorthaired/Wire-Haired Dachshund) Inheritance Autosomal recessive Sequencing (Beagle, Golden Retriever) Duration 1 – 2 weeks (Australian Cattle Dog, Australian Shepherd, Chinese Breed Beagle, Dachshund (Dackel), Golden Retriever, Long-Haired Crested Dog, Chihuahua, Dachshund, Saluki) Dachshund, Shorthaired Dachshund, Wire-Haired Dachshund 3 – 5 days (American Bulldog, Border Collie, English Setter, Gordon Inheritance Autosomal recessive Setter, Tibetan Terrier) Duration 2 – 3 days (Shorthaired/Wire-Haired Dachshund) Note NCL is a neurodegenerative disease, caused by lysosomal storage 1 – 2 weeks (Beagle, Golden Retriever) dysfunction. The clinical course includes increasing levels of agita- Note Osteogenesis imperfecta is characterised by extremely fragile bones tion and possible outbursts of aggression, hallucinations, hyperac- and teeth due to defects in the structure of collagen I. First symp- tivity and epileptic fits. Most animals lose their ability to coordinate toms appear already at young age. everyday muscular activities. As the extent of neurodegeneration increases, all affected dogs develop psychological abnormalities Paroxysmal Dyskinesia (PxD) and ataxia. The onset of the disease as well as its severity may vary strongly. Material EB 1 ml, buccal swab Method Sequencing Breed Irish Soft Coated Wheaten Terrier Inheritance Autosomal recessive Duration 1 – 2 weeks 294 295 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Note Affected dogs suffer from episodes of sudden, automatic and Polycystic Kidney Disease (PKD) unpredictable movements of especially the hindlimbs, known as hyperkinesia. These attacks last several minutes to hours and occur Material EB 1 ml, buccal swab about 10 times a day. First symptoms occur at the age of about two Method Sequencing years and worsen with age. Breed Bull Terrier Inheritance Autosomal dominant Duration 1 – 2 weeks Persistent Müllerian Duct Syndrome (PMDS) Note PKD is characterised by cysts in the liver, the pancreas and bilateral Material EB 1 ml, buccal swab renal cysts, ultimately leading to renal failure and death. Method Sequencing Breed Miniature Schnauzer Inheritance Autosomal recessive Postoperative Haemorrhage (P2Y12) Duration 1 – 2 weeks Material EB 1 ml, buccal swab Note Incomplete regression of the Mullerian duct during sex-differentiation Method Sequencing in the male dog. Normally, external genitalia are fully developed. Breed Great Swiss Mountain Dog Further symptoms include undescended testes, infertility and some- Inheritance Autosomal dominant times tumours. Duration 1 – 2 weeks Note A mutation found in the P2Y12-gene causes a severe bleeding Phosphofructokinase Deficiency (PFKD) disorder in Great Swiss Mountain Dog. Severe bleeding occurs after injuries or during surgery for the first time and ended fatal in many Material EB 1 ml, buccal swab known cases. The genetic test is a diagnostic tool to pre-emptively Method Sequencing check a dog for this predisposition for severe bleeding. Breed American Cocker Spaniel, English Springer Spaniel, German Spani- el, Whippet Inheritance Autosomal recessive Prekallikrein Deficiency (KLK) Duration 1 – 2 weeks Material EB 1 ml, buccal swab Note Affected dogs display the following intermittent, clinical signs: Method Sequencing weakness, exercise intolerance, muscle cramps, anaemia, jaundice Breed Shih Tzu and dark-coloured urine. Inheritance Autosomal recessive Duration 1 – 2 weeks Pituitary Dwarfism Note KLK leads to loss of prekallikrein function, a member of the coagula- tion cascade. Affected dogs show no or little symptoms of prolon- Material EB 1 ml, buccal swab ged bleeding. In combination with other defects in the coagulation Method FLP cascade (factor VII, VIII and IX deficiency), affected dogs did exhibit Breed Czechoslovakian Wolfdog, German Shepherd, Saarloos Wolfhond stronger bleeding, though. Inheritance Autosomal recessive Duration About 1 week Primary Ciliary Dyskinesia (PCD) Note Pituitary dwarfism produces perfectly proportioned but miniaturized dogs. It is caused by a gene defect, which leads to dysfunction of Material EB 1 ml, buccal swab the pituitary gland. As a result, lower amounts of growth hormones Method TaqMan SNP assay (Old English Sheepdog) and thyroxine reach the blood. Hence, affected dogs stop to grow Sequencing (Alaskan Malamute) between week three to eight of their lives. Breed Alaskan Malamute, Old English Sheepdog Inheritance Autosomal recessive Duration 3 – 5 days (Old English Sheepdog) 1 – 2 weeks (Alaskan Malamute) 296 297 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Note This syndrome is characterised by recurrent infections of the respira- Note A primary open angle glaucoma is a genetic dysfunction of the con- tory tract as well as reduced male fertility. Approximately 50% of the nective tissue in the eye. Thereby, the aqueous fluid is blocked and affected patients develop situs inversus (Kartagener syndrome). the pressure of the eyeball rises. Ultimately, the optic nerve and the retina are damaged leading from partial to complete blindness. The Primary Hyperoxaluria (PH) onset of the disease varies between breeds. Material EB 1 ml, buccal swab Method Sequencing Primary Open Angle Glaucoma and Lens Luxation (POAG/PLL) Breed Coton de Tuléar Material EB 1 ml, buccal swab Inheritance Autosomal recessive Method TaqMan SNP assay Duration 1 – 2 weeks Breed Shar-pei Note Accumulation of oxalate and formation of calcium oxalate crystals Inheritance Autosomal recessive in the urinary organs are a result of this disturbance. The resulting Duration 3 – 5 days crystals also accumulate in the kidney tissue and can lead to Note A genetic dysfunction of the connective tissue in the eye leads to decreased renal function. glaucoma (POAG) and often to lens luxation (PLL). POAG can lead to blindness. Primary Lens Luxation (PLL) Usually, affected dogs show first symptoms at the age of 4 – 6 years. Material EB 1 ml, buccal swab Method TaqMan SNP assay Progressive Retinal Atrophy (PRA) Breed American Eskimo Dog, American Hairless Terrier, Australian Cattle Progressive retinal atrophy (PRA) is a leading hereditary cause of blindness in pedigree Dog, Chinese Crested Dog, Danish-Swedish Farmdog, Fox Terrier, dogs as is its counterpart retinitis pigmentosa (RP) in humans. The onset of the German Hunting Terrier, Jack Russell Terrier, Lakeland Terrier, diseases varies between different breeds but is often not diagnosed before the age of Lancashire Heeler, Lucas Terrier, Miniature Bull Terrier, Norfolk Terrier, six. The progressive retinal atrophy (PRA) is leading to a degeneration of the photo Norwich Terrier, Parson Russell Terrier, Patterdale Terrier, Pug Dog, receptor cells of the retina. In most forms of PRA initially, a loss of function of the rod Rat Terrier, Sealyham Terrier, Teddy Roosevelt Terrier, Tenterfield cells is observed proceeding in night blindness and decreased adaptation of vision. In Terrier, Tibetan Terrier, Toy Fox Terrier, Volpino Italiano, Welsh Terrier, the final stage, degeneration of the cone cells results in complete blindness. The clinical Westfalen Terrier, Yorkshire Terrier and ophthalmologic signs of all forms are similar. Affected dogs suffer from bilateral Inheritance Autosomal recessive; it is estimated that about 2 – 20% of the carri- mydriasis, the reflection of the tapetum lucidum is increased and the retinal vascular ers will develop PLL. network appears atrophic. Fundus changes are bilateral and symmetrical. Duration 3 – 5 days Note Affected dogs have painful glaucomas and blindness due to a dis Bas-PRA1 location of the lens. Material EB 1 ml, buccal swab Method Sequencing Breed Basenji Primary Open Angle Glaucoma (POAG) Inheritance Autosomal recessive Material EB 1 ml, buccal swab Duration 1 – 2 weeks Method Sequencing Note The PRA form which can be explained by the investigated mutation Breed Basset Fauve de Bretagne, Basset Hound, Beagle, Norwegian has an onset of about 5 years. The known genetic variant does not Elkhound explain all PRA cases. So, one assumes that at least one further Inheritance Autosomal recessive mutation is involved in PRA genesis. Duration 1 – 2 weeks 298 299 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS CNGA1-PRA crd1-PRA Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay Method Sequencing Breed Shetland Sheepdog (Sheltie) Breed American Staffordshire Terrier Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 3 – 5 days Duration 1 – 2 weeks Note First symptoms of PRA affected Shetland Sheepdogs can be diag- Note see crd-PRA nosed at about two years of age. Another eye-disease in this breed, the so-called slow progressing retinopathy (SRP) exhibits similar crd2-PRA symptoms initially. Ophthalmologic differentiation is possible through Material EB 1 ml, buccal swab ERG only at that stage. Apart from the mutation in the CNGA1-gene Method Sequencing additional causative mutations for PRA might exist in Shetland Breed American Pitbull Terrier Sheepdogs. Inheritance Autosomal recessive Duration 1 – 2 weeks cord1-PRA Note See crd-PRA. Material EB 1 ml, buccal swab Method FLP Dominant PRA Breed Curly Coated Retriever, Dachshund, English Springer Spaniel, Material EB 1 ml, buccal swab Long-Haired Dachshund, Shorthaired Dachshund, Wire-Haired Method Sequencing Dachshund Breed Bullmastiff, Mastiff Inheritance Autosomal recessive Inheritance Autosomal dominant Duration About one week Duration 1 – 2 weeks Note In contrast to rod-cone dystrophies, where rod cells are affected first and the following degeneration of the cone cells results in complete g-PRA blindness of the dog, cone-rod dystrophies are characterised by the Material EB 1 ml, buccal swab relatively early loss of cone photoreceptors. Method Sequencing The first ophthalmoscopic signs may appear at the age of six Breed Schapendoes months. However, some genetically affected dogs never develop Inheritance Autosomal recessive clinical symptoms during their life. Duration 1 – 2 weeks crd-PRA GR-PRA1 and GR-PRA2 Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method FLP Method TaqMan SNP assay Breed Wire-Haired Dachshund Breed Golden Retriever Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 3 – 5 days Note The crd-PRA (cone-rod dystrophy) is an autosomal recessive Note The onset of the diseases varies but is often not diagnosed before inherited photoreceptor disease caused by the predominant loss the age of 6. of cone function. The earliest ophthalmoscopic signs appear at about six months of age. The complete manifestation of the disease (complete day blindness) occurs at an age of around 1 to 2. 300 301 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS pap-PRA1 rcd2-PRA Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay Method FLP Breed Papillon, Phalène Breed Collie (rough/smooth) Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 3 – 5 days Duration About 1 week Note The PRA form in Papillons and Phalènes show a wide range in age of onset maybe due to different genetic background or environ rcd3-PRA mental influences. The known genetic variant explains about 70% Material EB 1 ml, buccal swab of PRA affected dogs. So, one assumes that at least one further Method Sequencing mutation is involved in PRA genesis. Breed Chinese Crested Dog, Pomeranian, Welsh Corgi Inheritance Autosomal recessive prcd-PRA* (Optigen or partner laboratory) Duration 1 – 2 weeks Material EB 1 ml, buccal swab Method Partner laboratory rcd4-PRA Breed American Cocker Spaniel, American Eskimo Dog, Australian Material EB 1 ml, buccal swab Cattle Dog, Australian Shepherd, Australian Silky Terrier, Australian Method Sequencing Stumpy Tail Cattle Dog, Bearded Collie, Bolognese, Bolonka Breed Australian Cattle Dog, English Setter, Gordon Setter, Irish Red and Zwetna, Chesapeake Bay Retriever, Chihuahua, Chinese Crested White Setter, Irish Setter, Miniature Poodle, Old Danish Pointing Dog, Dog, English Cocker Spaniel, English Shepherd, Entlebucher Polish Lowland Sheepdog (PON), Poodle, Small Munsterlander, Mountain Dog, Finnish Lapphund, German Spitz, Giant Spitz, Tatra Shepherd Dog, Tibetan Terrier, Toy Poodle Gigant Schnauzer, Golden Retriever, Karelian Bear Dog, Kuvasz, Inheritance Autosomal recessive Labradoodle, Labrador Retriever, Lagotto Romagnolo, Lapponian Duration 1 – 2 weeks Herder, Markiesje, Miniature Australian Shepherd, Miniature Poodle, Norwegian Elkhound , Nova Scotia Duck Tolling Retriever, Poodle, Typ A-PRA* (Optigen) Portuguese Water Dog, Schipperke, Spanish Water Dog, Swedish Material EB 1 ml, buccal swab Lapphund, Toy Poodle, Wäller, Yorkshire Terrier Method Partner laboratory Inheritance Autosomal recessive Breed Miniature Schnauzer Duration 1 – 2 weeks (partner laboratory) Inheritance Autosomal recessive 4 – 6 weeks (Optigen) Duration 3 – 4 weeks Note The type A-PRA occurs very rarely. Therefore, the genetic test could rcd1-PRA be especially reasonable in the case of a clinical diagnosis of PRA. Material EB 1 ml, buccal swab Method Sequencing Type B-PRA* Optigen Breed Irish Red and White Setter, Irish Setter Material EB 1 ml, buccal swab Inheritance Autosomal recessive Method Partner laboratory Duration 1 – 2 weeks Breed Miniature Schnauzer Inheritance Autosomal recessive rcd1a-PRA Duration 3 – 4 weeks Material EB 1 ml, buccal swab Method Sequencing Breed Sloughi Inheritance Autosomal recessive Duration 1 – 2 weeks 302 303 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS XL-PRA Raine Syndrome Material EB 1 ml, buccal swab Method Sequencing Material EB 1 ml, buccal swab Breed Husky, Samoyed Method TaqMan SNP assay Inheritance X-chromosomal recessive Breed Border Collie Duration 1 – 2 weeks Inheritance Autosomal recessive Duration 3 – 5 days Note First symptoms usually occur at the age of three to five years. Note Affected dogs exhibit severe tooth wear resulting in pulpitis and the loss of most teeth. These symptoms result by hypomineralisation of Protein Losing Nephropathie (PLN) the teeth and weakened enamel. Skeletal bones can also be found Material EB 1 ml, buccal swab to be less mineralised in these dogs. Method TaqMan SNP assay Breed Irish Soft Coated Wheaten Terrier Renal Cystadenocarcinoma and Nodular Fibrosis (RCND) Inheritance Heterozygous dogs possess medium risk, homozygous affected dogs possess high risk to develop symptoms of the disease. Material EB 1 ml, buccal swab Duration 3 – 5 days Method TaqMan SNP assay Breed German Shepherd Note Hereditary PLN in Soft Coated Wheaten Terrier starts with hidden Inheritance Autosomal dominant proteinuria in midlife. The disease can proceed mildly and stay stea- Duration 3 – 5 days dy for years. In some cases, severe complications like kidney failure or thromboses occur. Note RCND causes bilateral, multifocal tumours in the kidney, uterine leiomyomas in female dogs and nodules in the skin consisting of dense collagen fibres. This form of cancer is inherited in a dominant Pyruvate Dehydrogenase Phosphatase 1 Deficiency (PDP1) pattern and seems to have a homozygous lethal effect on most Material EB 1 ml, buccal swab dogs so that these fetuses often die before birth. Method Sequencing Breed Clumber Spaniel, Sussex Spaniel Retinal Dysplasia* (OSD) (Optigen or partner laboratory) Inheritance Autosomal recessive Duration 1 – 2 weeks Material EB 1 ml, buccal swab Method Partner laboratory Note The disease is characterised by exercise intolerance and post- Breed Labrador Retriever, Samoyed exercise collapse. There may also be neurological symptoms. Inheritance Autosomal dominant with incomplete penetrance Duration 1 – 2 weeks (partner laboratory) Pyruvate Kinase Deficiency (PK) 4 – 6 weeks (Optigen) Material EB 1 ml, buccal swab Note Retinal folds, also called retinal dysplasia (RD), is observed in many Method Sequencing dog breeds without any pathological consequence. However, in Breed Basenji, Beagle, Cairn Terrier, Labrador Retriever, Pug Dog, West the Labrador Retriever breed as well as in Samoyeds, retinal folds Highland White Terrier can be associated with a more severe syndrome, the oculoskeletal Inheritance Autosomal recessive dysplasia (OSD). Clinical signs of OSD are skeletal malformations Duration 1 – 2 weeks comprising shortened limbs (dwarfism) as well as blindness at early Note In dogs, the anaemia caused by this enzyme deficiency in erythro age. cytes is always severe. Also associated with the disease is a progressive myelofibrosis and osteosclerosis of unknown aetiology. 304 305 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Sensory Neuropathy (SN) Skeletal Dysplasia 2 (Dwarfism) (SD2) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method FLP Method TaqMan SNP assay Breed Border Collie Breed Labrador Retriever Inheritance Autosomal recessive Inheritance Autosomal recessive Duration About 1 week Duration 1 – 2 weeks Note Sensory neuropathy in the Border Collie is a severe neurological Note Skeletal dysplasia 2 causes an early halt in growth of long bones. disorder caused by the degeneration of sensory and, to a lesser In contrast to other forms of dwarfism (pituitary dwarfism), the result extent, motor nerve cells. Clinical signs start between 2 and 7 are „disproportioned“ dogs with shortened front limbs and raising months of age and include progressive proprioceptive ataxia with dorsal line. Torso length and depth is not altered. intermittent knuckling of the paws, hyperextension of the limbs, and self-mutilation wounds in the distal part of the limbs. In some cases, Spinal Dysraphism (NTD) autonomic signs such as urinary incontinence and, in the later stage, regurgitation has also been reported. Material EB 1 ml, buccal swab Method Sequencing Breed Weimaraner Severe Combined Immuno Deficiency (SCID) Inheritance Autosomal recessive Material EB 1 ml, buccal swab Duration 1 – 2 weeks Method Sequencing Note Neural tube defects result from abnormal closure or development Breed Frisian Water Dog, Jack Russell Terrier, Parson Russell Terrier of the neuronal tube during embryogenesis. The spinal dysraphism Inheritance Autosomal recessive is a non-progressive ataxia and is characterised by the following Duration 1 – 2 weeks symptoms: abnormal hair streams along the back, kinked tails, Note SCID is associated with very low levels of immunoglobulins and scoliosis in the lumbar spinal region, paraparesis, “bunny-hopping”, lymphocytes and therefore causes serious failing in cellular and crouched stance. humoral immunity. Affected dogs show increased susceptibility for viral and bacterial pathogens and die of opportunistic infection at the Spinocerebellar Ataxia (SCA) age of 8 – 12 weeks. Material EB 1 ml, buccal swab Method TaqMan SNP assay Shar Pei Autoinflammatory Disease (SPAID) Breed Fox Terrier, Jack Russell Terrier, Parson Russell Terrier, Tenterfield Material EB 1 ml, buccal swab Terrier, Toy Fox Terrier Method TaqMan SNP assay Inheritance Autosomal recessive Breed Shar-pei Duration 3 – 5 days Inheritance Autosomal dominant with incomplete penetrance Note Affected dogs will show first indications of the ataxia at the age Duration 3 – 5 days of 3 – 6 months. First clinical signs are loss of balance and minor Note In Shar Pei, signs of inflammation including fever and arthritis incoordination of gait while later symptoms can be a progressive are known to be related with a breed-specific predisposition for incoordination or a complete loss of mobility. a syndrome termed Shar-Pei Autoinflammatory Disease (SPAID). SPAID could be characterised by the following varying symptoms: arthritis, dermatitis, otitis, systemic amyloidosis, erythema in the region of the wrinkles, thickened and pasty skin, eye inflammation and recurrent intestinal inflammations. First clinical signs of the disease usually appear at the age of 1 to 6 years. 306 307 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Spondylocostal Dysostosis (Comma Defect) Subacute Necrotising Encephalopathy (SNE) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method FLP Breed Miniature Schnauzer Breed Yorkshire Terrier Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 1 – 2 weeks Note The spondylocostal dysostosis (comma defect) is characterised Note SNE in Yorkshire Terriers usually manifests during the first year of life. mainly by dysfunction of segmentation of the spine and ribs. Typical symptoms are: abnormal walking, ataxia and spasticity as Newborns exhibit a disproportional dwarfism. They also have a well as visual and perceptual disorders. Affected dogs have to be prominent forehead, an expansive occipital and malformations of the euthanised. extremities. The malformation of the ribcage lowers chest-volume, which leads to decreased respiratory function. Thrombopathia Material EB 1 ml, buccal swab Spongy Degeneration with Cerebellar Ataxia (SDCA1 and 2) Method TaqMan SNP assay (Basset Hound) Material EB 1 ml, buccal swab Sequencing (Landseer) Method TaqMan SNP assay (SDCA1) Breed Basset Hound, Landseer FLP (SDCA2) Inheritance Autosomal recessive Breed Belgian Shepherd, Dutch Shepherd Duration 3 – 5 days (Basset) Inheritance Autosomal recessive 1 – 2 weeks (Landseer) Duration 3 – 5 days (SDCA1) Note The thrombocytes of dogs which suffer from this kind of hereditary About 1 week (SDCA2) thrombopathia do not respond normally to activation signals. Note SDCA is a neurodegenerative disease. Affected animals show a tendency to bruising and haemorrhages. The puppies with cerebellar dysfunction have early onset of clinical signs (5 – 8 weeks of age). They show a wide-based ataxic gait, Trapped Neutrophil Syndrome (TNS) particularly obvious in the hind limbs. Other clinical signs were stumbling, staggering, intention tremor, bunny hopping, muscle Material EB 1 ml, buccal swab spasms, as well as balance loss and falling. SDCA is a progressive Method TaqMan SNP assay disease and the affected dogs are usually euthanised after 12 weeks Breed Border Collie at the latest. Inheritance Autosomal recessive Duration 3 – 5 days Startle Disease Note In TNS affected dogs, neutrophils are produced by the bone marrow, but lack the ability to reach the blood stream. Therefore, affected Material EB 1 ml, buccal swab puppies possess a weak immune system. The variable symptoms Method FLP observed depend on the respective infection. Equally, starting point Breed Irish Wolfhound and severity of the TNS-disease vary, with most of the dogs dying Inheritance Autosomal recessive before the fifth month of their lives. Duration 1 – 2 weeks Note First symptoms appear 5 – 7 days after birth. Affected dogs show extensor rigidity and tremor evoked by handling. Symptoms cease when dogs are relaxed or sleeping. The puppies are unable to stand and show rigid extended posture in all four limbs. Affected puppies have to be euthanised. Laboklin owns the exclusive license to perform this genetic test. 308 309 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS van den Ende-Gupta Syndrome (VDEGS) von-Willebrand Disease Type II (vWD 2) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay Method TaqMan SNP assay Breed Fox Terrier, Toy Fox Terrier Breed German Shorthaired Pointer, German Wirehaired Pointer Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 3 – 5 days Duration 3 – 5 days Note All affected dogs exhibit a prominent underbite with short maxilla. Note Clinical symptoms of vWD are highly heterogeneous from mild Additionally, bones are often not mineralised and luxation of the el bleeding to severe life-threatening blood loss. bow or patella as well as swollen knee joints can be found regularly. von Willebrand Disease Type III (vWD3) Vitamin D Dependent Rickets (VDR) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay (Scottish Terrier, Shetland Sheepdog) Method Sequencing Sequencing (Kooikerhondje) Breed Pomeranian Breed Kooikerhondje, Scottish Terrier, Shetland Sheepdog (Sheltie) Inheritance Unknown Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 3 – 5 days (Scottish Terrier, Shetland Sheepdog) Note The hereditary form of vitamin D-dependent rickets type II is caused 1 – 2 weeks (Kooikerhondje) by a defect in the vitamin D receptor gene VDR. As a consequence, Note Clinical symptoms of vWD are highly heterogeneous from mild calcium cannot be absorbed intestinally, which results in skeletal bleeding to severe life-threatening blood loss. malformation and hypomineralisation of bones during growth at a young age. Because the VDR gene is also involved in the hair Warburg Micro Syndrome 1 (WARBM1) growth cycle, alopecia may occur as well. Material EB 1 ml, buccal swab Method Sequencing von-Willebrand Disease Type I (vWD I) Breed Husky Material EB 1 ml, buccal swab Inheritance Autosomal recessive Method TaqMan SNP assay Duration 1 – 2 weeks Breed Bernese Mountain Dog, Coton de Tuléar, Dobermann, Drentse Note Symptoms include developmental defects and polyneuropathy. Patrijshond, German Pinscher, Kerry Blue Terrier, Kromfohrländer, Affected dogs exhibit polyneuropathy with ocular abnormalities and Labradoodle, Manchester Terrier, Papillon, Poodle, Stabijhoun, neuronal vacuolation. Additionally, a progressive severe ataxia deve- Welsh Corgi Pembroke lops, which leads to euthanasia of the puppies at an age between 8 Inheritance Autosomal dominant with variable penetrance to 16 months. Duration 3 – 5 days Note Clinical symptoms of vWD are highly heterogeneous from mild bleeding to severe life-threatening blood loss. 310 311 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS X-chromosomal Severe Immunodeficiency (X-SCID) A-locus (alleles: Ay, Aw, at, a) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method TaqMan SNP assay + FLP Breed Basset Hound, Welsh Corgi Cardigan, Welsh Corgi Pembroke Breed All Inheritance X-chromosomal recessive Duration About one week Duration 1 – 2 weeks Note Affected dogs suffer from developmental disorder, increased B-locus (brown, chocolate, liver(nose)) responsiveness for viral and bacterial pathogens and degeneration Material EB 1 ml, buccal swab of peripheral lymph nodes. The dogs die at the age of four months. Method TaqMan SNP assay Breed All X-linked Myopathy (XL-MTM) Duration 3 – 5 days Material EB 1 ml, buccal swab Method Sequencing C-locus (albino) Breed Labrador Retriever, Rottweiler Material EB 1 ml, buccal swab Inheritance X-chromosomal recessive Method Sequencing Duration 1 – 2 weeks Breed German Spitz, Giant Spitz, Keeshond, Lhasa Apso, Pekingese, Note XL-MTM is a hereditary disorder that affects the skeletal muscles Pomeranian in the body. Clinical signs for this disease can be seen from birth. Duration 1 – 2 weeks Symptoms are strong hypotonia, muscle atrophy and progressive weakening of the hind limbs. This is usually accompanied by breathing difficulties, which can lead to death by suffocation. 20.2.2 Coat Colour/Coat Structure Dog Mammals inherit two pigments that are the basis for hair colour: eumelanin (black) and pheomelanin (red or yellow). The rich colour varieties seen among dog breeds arise due to genes controlling the amount, extent, and distribution of these two colour pig- ments. Involved in the production of these pigments in many species including dogs is melanocortin 1 receptor (MC1R) gene which is also called extension. Other genes modify the distribution of these pigments to produce the variety of colours and pat- terns found in the domestic dog. The brown gene, tyrosinase-related protein 1 (TYRP1) gene, is a modifier that alters black pigment to brown but does not affect red pigment. Other genes involved in canine coat colour include Agouti (ASIP) which organises the distribution of black and red pigments and Dilute (MLPH) which dilutes black to grey and red to cream. A lot of other genes specific to some breeds of dogs exist that add white patterns and dilute colours. Below expanded descriptions of the genetic tests offered by LABOKLIN for dog coat colour can be found. 312 313 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS 314 315 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog LABORATORY FOR CLINICAL DIAGNOSTICS Coat Length (long or short hair) EG-locus (domino, grizzle) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay Method Sequencing Breed All Breed Afghan Hound, Barzoi, Saluki Duration 3 – 5 days Duration 1 – 2 weeks Coat Length II (long or short hair) EH-locus (sable) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method TaqMan SNP assay Breed Afghan Hound, Akita, Alaskan Malamute, American Akita, Chow Breed American Cocker Spaniel, English Cocker Spaniel Chow, Eurasien, French Bulldog, Husky, Samoyed, Shiba Duration 3 – 5 days Duration 1 – 2 weeks EM-locus (melanistic mask) Curly (curled hair) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay Method TaqMan SNP assay Breed All Breed All Duration 3 – 5 days Duration 3 – 5 days Furnishing (wire hair) D-locus d1 (dilution) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method TaqMan SNP assay Breed All Breed All Duration 1 – 2 weeks Duration 3 – 5 days H-locus (harlequin) D-locus d2 (dilution) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay Method Sequencing Breed Great Dane Breed Chow Chow, Sloughi, Thai Ridgeback Dog Duration 3 – 5 days Duration 1 – 2 weeks Note Harlequin is a typical coat colour of Great Danes, which show black patterns on white background. The dominant H-allele is lethal when E-locus (yellow, lemon, red, cream, apricot) found homozygous (H/H). Affected puppies die before birth. Material EB 1 ml, buccal swab Method TaqMan SNP assay Hairlessness (powderpuff) Breed All Material EB 1 ml, buccal swab Duration 3 – 5 days Method FLP Note In Australian Shepherds, Border Collies and other herding dogs, the Breed Chinese Crested Dog, Mexican Hairless Dog (Xoloitzcuintle), colour variant “red” is detected by genetic testing of the B-locus. Peruvian Hairless Dog Duration 1 – 2 weeks Note Dogs which are homozygous for the mutation die as embryo during gestation. 316 317 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Dog 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cat LABORATORY FOR CLINICAL DIAGNOSTICS Improper Coat S-locus (Piebald, White Spotting) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method FLP Breed Portuguese Water Dog Breed All Duration 1 – 2 weeks Duration 1 – 2 weeks Note White spotting in dogs is mostly caused by variations of MITF. K-locus (only the allele KB) Depending on a short interspersed nucleotide element (SINE), the Material EB 1 ml, buccal swab dog is spotted or not. Del is the dominant allele and causes solid or Method Sequencing single-coloured dogs (genotype del/del). Breed All Deafness could be associated with white spotting in the case of a Duration 1 – 2 weeks very severe form of white spotting and/or a combination of other diluting factors (dilution, merle). Deafness especially occurs in animals with white spots at the head and the ears. M-locus* (alleles: Mh, M, Ma, Mc, m) Material EB 1 ml, buccal swab Method Partner laboratory 20.3 Cat Breed All Duration 1 – 2 weeks 20.3.1 Hereditary Diseases Note Merle is a coat pattern characterised by dark patches intermingled with diluted pigment. This trait is inherited in an autosomal, incom- Alpha-Mannosidosis (AMD) pletely dominant fashion. Additionally, a shortened version of the Material EB 1 ml, buccal swab causative M variant exists on the M-locus. This so-called cryptic Method Sequencing merle (Mc) has no influence on the coat colour itself. Dogs homo- Breed Persian zygous for merle (M/M) are also known as double merles and are Inheritance Autosomal recessive predominantly white. In all breeds, the double merle genotype can Duration 1 – 2 weeks be sublethal and is associated with multiple auditory and ophthal- mologic abnormalities. For these reasons, merle-to-merle breedings Note Alpha-mannosidosis is a lysosomal storage disease, symptoms are strongly prohibited to avoid pain breeding. include malformation in bone structure and severe neurological symptoms such as ataxia, tremor and limited vision. Affected cats usually die after birth or in the first months of life. Panda White Spotting Material EB 1 ml, buccal swab Autoimmune Lymphoproliferative Syndrome (ALPS) Method Sequencing Breed German Shepherd Material EB 1 ml, buccal swab Duration 1 – 2 weeks Method Sequencing Breed British Shorthair Note The Panda white spotting is inherited in an autosomal dominant Inheritance Autosomal recessive mode, the homozygous mutation is embryonic lethal. Duration 1 – 2 weeks Note ALPS is a rare non-neoplastic lymphoproliferative disease charac- Saddle-tan terised by lymphadenopathy and splenomegalie with autoimmune Material EB 1 ml, buccal swab cytopenia. ALPS has an early onset, affected cats show symptoms Method Sequencing during the first weeks with a rapidly progressive clinical course. So Breed Basset Hound, Welsh Corgi far, the mutation was exclusively detected in British Shorthair cats Duration 1 – 2 weeks from Australia and New Zealand. 318 319 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cat 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cat LABORATORY FOR CLINICAL DIAGNOSTICS Congenital Myasthenic Syndrome (CMS) Genetic Blood Group Determination Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method TaqMan SNP assay Breed Devon Rex, Sphynx Breed All except European Shorthair Inheritance Autosomal recessive Duration 3 – 5 days Duration 1 – 2 weeks Note Neonatal isoerythrolysis occurs when a female cat with blood type B Note Symptoms of the disease correlate to human CMS and are is bred to an A-type male cat and the A-type kitten absorb the anti-A described as generalised amyosthenia particularly following stress antibodies from the breast milk. This haemolytic disease can also be or agitation. First symptoms appear at three weeks of age. Affected lethal. The genetic determination of the blood types in cats enables cats often exhibit a “squirrel”-like posture and rest their front paws the genetic differentiation of the serologically determined blood on elevated objects. Almost all cats with CMS die during their first group before breeding (see also Chapter 3.3). two years of life. Glycogen Storage Disease Type IV (GSD4) Cystinuria (CysK) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method FLP Method Sequencing Breed Norwegian Forest Cat Breed All Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 1 – 2 weeks Note Most affected kittens die at or soon after birth, presumably due to Note Cystinuria is an inherited disorder caused by a defective transport hyperglycaemia. Survivors of the perinatal period appear clinically of the amino acid cystine in the kidney tubules. Cats with Cystinuria normal until onset of progressive neuromuscular degeneration at 5 do not properly reabsorb the cystine (and a few other amino acids) months of age that eventually leads to death. in the kidney tubules, causing the urine to contain abnormally high levels of cystine. Cystine is insoluble in neutral pH or acidic urine, Head Defect so excess urinary cystine results in the formation of crystals, which in turn can lead to formation of cystine calculi (stones) in the kidney Material EB 1 ml, buccal swab and/or the bladder. Method Sequencing Breed Burmese Inheritance Autosomal recessive Gangliosidosis (GM1/GM2) Duration 1 – 2 weeks Material EB 1 ml, buccal swab Note One copy of the mutation does not cause the craniofacial defect but Method Sequencing may produce a shortened facial structure (brachycephaly). Cats with Breed Balinese, Burmese, Burmilla, Javanese, Korat, Oriental Shorthair two copies of the mutation have the severe craniofacial defect that is (OSH), Peterbald, Siamese, Thai, Tonkinese incompatible with life. Inheritance Autosomal recessive Duration 1 – 2 weeks Hypertrophic Cardiomyopathy (HCM) Note There are two types of gangliosidoses, GM1 and GM2 gangliosidosis. Affected kittens have head tremors at the beginning Material EB 1 ml, buccal swab followed by impaired coordination of leg movements which Method TaqMan SNP assay eventually lead to paralysis. Breed Maine Coon, Ragdoll In GM2 (Burmese, Korat), symptoms normally occur earlier (at about Inheritance Autosomal dominant 2 months of age) and worsen faster. In GM1 (Siamese, Korat), the Duration 3 – 5 days neurologic symptoms start a little later (about 3 months of age) and progress slower. 320 321 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cat 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cat LABORATORY FOR CLINICAL DIAGNOSTICS Note HCM is characterised by an increased left ventricular mass due to nervous system as well as dwarfism. First clinical signs of the severe an increase in wall thickness of the heart, homozygous animals run a phenotype appear after only few weeks of life. significantly increased risk of developing HCM compared to healthy animals. Heterozygous carriers only run a slightly increased risk. Mucopolysaccharidosis Type VII (MPS7) Material EB 1 ml, buccal swab Hypokalemia Method Sequencing Material EB 1 ml, buccal swab Breed All Method TaqMan SNP assay Inheritance Autosomal recessive Breed Australian Mist, Burmese, Burmilla, Cornish Rex, Devon Rex, Duration 1 – 2 weeks Singapura, Sphynx, Tonkinese Note Mucopolysaccharidosis Type VII is a rare lysosomal storage disease Inheritance Autosomal recessive caused by β-glucuronidase deficiency. Affected cats show growth Duration 3 – 5 days retardation, corneal clouding, dental eruption and multiple skeletal Note Burmese hypokalemia, also known as familial episodic hypo abnormalities. First anatomical abnormalities are seen at the age of kalaemic polymyopathy, is characterised by episodes of skeletal about two months. muscle weakness. As a result, affected cats may show problems with walking and holding their head correctly. Myotonia Congenita Material EB 1 ml, buccal swab Hypotrichosis and Short Life Expectancy Method Sequencing Material EB 1 ml, buccal swab Breed All Method Sequencing Inheritance Autosomal recessive Breed Birman (Sacred cat of Burma) Duration 1 – 2 weeks Inheritance Autosomal recessive Note Myotonia congenita (MC) is a skeletal muscle channelopathy Duration 1 – 2 weeks characterised by inability of the muscle to relax after voluntary Note Affected kitten are either born bald or have a thin downy coat that contraction. Affected cats have a protruding tongue, limited range falls out within a week of birth. Regrowth of a thin coat may occur of jaw motion and drooling with prominent neck and proximal limb within the first two months in some animals. In addition, oily and musculature. Additionally, these cats show a short-strided gait, crusty skin in the facial area and anomalies of the claws, tongue and problems while swallowing and excessive salivation. beard are other clinical symptoms. The disease also leads to stillbir- ths and early death cases in kittens in the first thirteen weeks of life Osteochondrodysplasia (OCD) due to impaired immune response. Material EB 1 ml, buccal swab Mucopolysaccharidosis Type VI (MPS6) Method Sequencing Material EB 1 ml, buccal swab Breed Scottish Fold Longhair, Scottish Fold Shorthair Method Sequencing Inheritance Autosomal dominant Breed Balinese, Birman (Sacred cat of Burma), European Shorthair, Duration 1 – 2 weeks Javanese, Oriental Shorthair (OSH), Peterbald, Siamese, Thai, Note The mutation in the TRPV4 gene leads to the phenotype of Tonkinese Scottish fold cats with the characteristic ear shape. Furthermore, Inheritance Autosomal recessive this mutation causes osteochondrodysplasia in this breed with Duration 1 – 2 weeks malformation in bones and joints of the distal limbs and tail. Note Mucopolysaccharidosis Type VI is a lysosomal storage disease Homozygous mutant cats seem to be more affected, therefore it is resulting in a clinically mild and a severe MPS VI phenotype, not recommended to pair Scottish fold cats with one another. characterised by severe disorders of the bone structure and the 322 323 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cat 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cat LABORATORY FOR CLINICAL DIAGNOSTICS pd-PRA Polycystic Kidney Disease (PKD) Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method TaqMan SNP assay Breed Birman (Sacred cat of Burma), British Longhair, British Shorthair Breed Birman (Sacred cat of Burma), British Longhair, British Shorthair (BSH), Chartreux, Exotic Shorthair, Kartäuser, Persian, Ragdoll, (BSH), Chartreux, Exotic Shorthair, Persian, Ragdoll, Russian Blue, Russian Blue, Scottish Fold Longhair, Scottish Fold Shorthair, Selkirk Scottish Fold Longhair, Scottish Fold Shorthair, Selkirk Rex Longhair, Rex Longhair, Selkirk Rex Shorthair, Turkish Angora Selkirk Rex Shorthair, Turkish Angora Inheritance Autosomal recessive Inheritance Autosomal dominant Duration 1 – 2 weeks Duration 3 – 5 days Note Onset of photoreceptor loss is around 5 weeks of age with severe Note PKD causes the formation of hepatic and pancreatic cysts as well as loss by 16 weeks of age. In affected cats, uncoordinated eye of fluid-filled renal cysts, often leading to renal failure. movement is often observed, as well as owner-reported increased eye-shine (tapetal reflectivity) as thinning of the retina progresses. Primary Congenital Glaucoma (PCG) rdAc-PRA Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method Sequencing Method TaqMan SNP assay Breed Siamese Breed Abyssinian, American Curl Longhair, American Curl Shorthair, Inheritance Autosomal recessive American Wirehair, Balinese, Bengal (Leopard cat), Cornish Rex, Duration 1 – 2 weeks Javanese, Munchkin, Ocicat, Oriental Shorthair (OSH), Peterbald, Note In general, a distinction is drawn between the primary glaucoma Siamese, Singapura, Somali, Thai, Tonkinese (congenital malformation of the eyes) and the secondary glaucoma Inheritance Autosomal recessive (caused by various eye diseases or infections). In domestic cats, Duration 3 – 5 days a mutation in LTBP2 causes primary congenital glaucoma (PCG). Note Affected cats have normal vision at birth. The age of onset of clinical Elevated intraocular pressure, globe enlargement and elongated symptoms is typically at the age of 1.5 – 2 years. At the end stage ciliary processes were consistently observed in all affected cats by 8 of disease complete photoreceptor degeneration and blindness is weeks of age. Varying degrees of optic nerve damage resulted by 6 observed, usually at the age of 3 – 5 years. months of age. rdy-PRA Progressive Retinal Atrophy (PRA) Material EB 1 ml, buccal swab Method Sequencing b-PRA Breed Abyssinian, Ocicat, Somali Material EB 1 ml, buccal swab Inheritance Autosomal dominant Method TaqMan SNP assay Duration 1 – 2 weeks Breed Bengal Note Cats carrying one copy of this mutation have retarded development Inheritance Autosomal recessive and degeneration of photoreceptor cells, which leads to early-onset Duration 3 – 5 days blindness by 7 weeks of age. Note Bengal progressive retinal atrophy causes the destruction of the photoreceptors in the retina. Loss of cells begins around 7 weeks of age and slowly progresses until the cat has very compromised vision by approximately 2 years of age. However, blindness develops at different rates in different cats. 324 325 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cat 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cat LABORATORY FOR CLINICAL DIAGNOSTICS Pyruvate Kinase Deficiency (PK) Coat Colour Dilution Material EB 1 ml, buccal swab Material EB 1 ml, buccal swab Method TaqMan SNP assay Method TaqMan SNP assay Breed Abyssinian, Bengal (Leopard cat), Egyptian Mau, European Breed All Shorthair, LaPerm Longhair, LaPerm Shorthair, Maine Coon, Duration 3 – 5 days Norwegian Forest Cat, Ocicat, Savannah, Siberian, Singapura, Somali, Turkish Angora Coat Colour Russet Inheritance Autosomal recessive Duration 3 – 5 days Material EB 1 ml, buccal swab Method Sequencing Note This deficiency manifests as a haemolytic anaemia of variable Breed Burmese severity with a strong regenerative response. The clinical signs Duration 1 – 2 weeks of disease reflect the anaemic status of the animal and include exercise intolerance, weakness, heart murmur and splenomegaly. Coat Colour Variant Agouti Spinal Muscular Atrophy (SMA) Material EB 1 ml, buccal swab Method TagMan SNP assay Material EB 1 ml, buccal swab Breed All Method FLP Duration 3 – 5 days Breed Maine Coon Inheritance Autosomal recessive Duration 1 week Coat Colour Variant Albino Note SMA is a disorder caused by death of spinal cord neurons that Material EB 1 ml, buccal swab activate skeletal muscles of the trunk and limbs. Loss of neurons Method Sequencing leads to muscle weakness and atrophy that first becomes apparent Breed All at 3 – 4 months of age. Duration 1 – 2 weeks Coat Colour Variant Charcoal 20.3.2 Coat Colour/Coat Structure Cat Material EB 1 ml, buccal swab Method Sequencing Coat Colour Amber Breed Bengal Duration 1 – 2 weeks Material EB 1 ml, buccal swab Method TaqMan SNP assay Breed Norwegian Forest Cat Coat Colour Variant Colourpoint (Siam/Mink/Burma) Duration 3 – 5 days Material EB 1 ml, buccal swab Method TaqMan SNP assay Coat Colour Brown (chocolate/cinnamon) Breed All except Bengal Duration 3 – 5 days Material EB 1 ml, buccal swab Method TaqMan SNP assay Breed All Duration 3 – 5 days 326 327 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cat 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Horse LABORATORY FOR CLINICAL DIAGNOSTICS Coat Colour Variant Dominant White/White Spotting Breed Quarab, Quarter Horse, Quarter pony Inheritance X-chromosomal recessive Material EB 1 ml, buccal swab Duration 1 – 2 weeks Method Sequencing Breed All Note AIS is a condition that results in the partial or complete inability of Duration 1 – 2 weeks the cell to respond to androgens, so the animal does not become masculinised. This results in XY (genetically male) horses with female phenotype (female external genitalia and internal testis) with Coat Colour Variant Snow male-like behaviour. These individuals are unable to reproduce. Material EB 1 ml, buccal swab Method Sequencing Cerebellar Abiotrophy (CA) Breed Bengal Duration 1 – 2 weeks Material EB 1 ml, hair roots Method TaqMan SNP assay Breed Anglo-Arabian, Arabian Horse, Shagya Arabian Coat Length (long or short hair) Inheritance Autosomal recessive Material EB 1 ml, buccal swab Duration 3 – 5 days Method Sequencing Note Foals affected with CA appear normal at birth. Around six weeks of Breed All age (although sometimes as late as four months), the disease causes Duration 1 – 2 weeks the death of neurons in the cerebellum, leading to head tremor and a lack of balance equilibrium (ataxia), among other neurological Coat Variant Curly deficits. Signs of CA are variable. Material EB 1 ml, buccal swab Method Sequencing Congenital Myotonia Breed Selkirk Rex Material EB 1 ml, hair roots Inheritance Autosomal dominant Method TaqMan SNP assay Duration 1 – 2 weeks Breed German Riding Pony, New Forest Pony Inheritance Autosomal recessive Coat Variant Sphynx/Devon Rex Duration 3 – 5 days Material EB 1 ml, buccal swab Note The first symptoms are recurrent episodes of recumbency and diffi Method Sequencing culty rising to its feet as a result of muscle stiffness. They occur during Breed Devon Rex, Sphynx the first weeks of age and usually increase in the following months. Inheritance Autosomal recessive Duration 1 – 2 weeks Dwarfism Material EB 1 ml, hair roots Method Sequencing 20.4 Horse Breed Friesian horse Inheritance Autosomal recessive 20.4.1 Hereditary Diseases Duration 1 – 2 weeks Note Dwarfism in Friesian horses is characterised by physeal growth Androgen Insensitivity Syndrome (AIS) retardation of limbs and ribs while the head and back appear normal. A striking feature of the condition is hyperextension of the Material EB 1 ml, 20 – 30 hair roots from mane or tail fetlock joints. Flexor tendon laxity, which is often seen in newborn Method Sequencing 328 329 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Horse 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Horse LABORATORY FOR CLINICAL DIAGNOSTICS foals of all breeds, fails to recover in dwarf foals and instead tends Note Clinical signs associated with GBED are: Abortion, dead or weak to increase further during aging. As a consequence, these dwarf foals; sudden death due to heart failure and/or seizures; tachypnoea Friesians develop an abnormal gait in which the limbs undergo due to weak skeletal muscle (diaphragm); generalised weakness, extreme outward rotation at the level of carpus and hocks. especially when getting up. The ribcage is abnormal in most cases with thickened and S-shaped costochondral junctions, leading to an inward protrusion Hereditary Equine Regional Dermal Asthenia (HERDA) of the chest at the level of Th10-16. Mature dwarfs have a head of the same size as unaffected horses, a broader chest with Material EB 1 ml, hair roots narrowing at the costochondral junction, a disproportionally long Method TaqMan SNP assay back and abnormally short limbs. The abdomen has a weak and Breed Appaloosa, Paint Horse, Quarab, Quarter Horse, Quarter pony rounded appearance, and the musculature over the body is poorly Inheritance Autosomal recessive developed. Duration 3 – 5 days Note The skin of affected horses is hyperextensible, scarred, and shows Equine Malignant Hyperthermia (EMH) severe lesions along the back. Material EB 1 ml, hair roots Method TaqMan SNP assay Hereditary Junctional Epidermolysis Bullosa (JEB) Breed Appaloosa, Paint Horse, Quarab, Quarter Horse, Quarter pony Material EB 1 ml, hair roots Inheritance Autosomal dominant Method Sequencing Duration 3 – 5 days Breed Ardennes Horse, Belgian Horse Note EMH is a life-threatening pharmacogenetic disorder of skeletal Inheritance Autosomal recessive muscle elicited by halogenated anaesthetics, depolarizing muscle Duration 1 – 2 weeks relaxants, and stress. Clinical and laboratory manifestations Note H-JEB is an inherited disease that causes moderate to severe include tachycardia, hyperthermia, muscle rigidity, rhabdomyolysis, blistering of the skin and mouth epithelia, and sloughing of hooves respiratory and metabolic acidosis, and electrolyte derangements. in newborn foals. Foal Immunodeficiency Syndrome (FIS) Hoof Wall Separation Disease (HWSD) Material EB 1 ml, hair roots Material EB 1 ml, hair roots Method Sequencing Method TaqMan SNP assay Breed Dales Pony, Fell Pony Breed Connemara Pony, German Riding Pony Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 3 – 5 days Note Foals with FIS appear to be normal at birth but within a few weeks Note Hoof wall separation disease (HWSD) is characterised by a hoof wall develop evidence of infection such as diarrhoea, pneumonia, etc. that easily breaks and cracks. The breaks and cracks begin to occur in The infections are resistant to treatment, and the foals die or are young ponies. Some cases are milder while others are more severe. euthanised before three months of age. Hydrocephalus Glycogen Branching Enzyme Deficiency (GBED) Material EB 1 ml, hair roots Material EB 1 ml, hair roots Method Sequencing Method TaqMan SNP assay Breed Friesian horse Breed Appaloosa, Paint Horse, Quarab, Quarter Horse, Quarter pony Inheritance Autosomal recessive Inheritance Autosomal recessive Duration 1 – 2 weeks Duration 3 – 5 days 330 331 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Horse 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Horse LABORATORY FOR CLINICAL DIAGNOSTICS Note Hydrocephalus in Friesian Horses often results in stillbirth of affected Lavender Foal Syndrome (LFS) foals and dystocia in dams. Material EB 1 ml, hair roots Method TaqMan SNP assay Hyperkalemic Periodic Paralysis (HYPP) Breed Arabian Horse Material EB 1 ml, hair roots Inheritance Autosomal recessive Method TaqMan SNP assay Duration 3 – 5 days Breed Appaloosa, Paint Horse, Quarab, Quarter Horse, Quarter pony Note Affected foals can display an array of neurological signs including Inheritance Autosomal dominant tetanic-like seizures, opisthotonus, stiff or paddling leg movements Duration 3 – 5 days and nystagmus. These neurologic impairments prevent the foal from Note Hyperkalaemic periodic paralysis has been reported in Quarter standing and nursing normally and, if not lethal on their own, are Horses (offspring of the stallion “Impressive”) and horses with often the cause for euthanasia. Quarter Horse blood (Appaloosas and Paints). The disease is characterised by intermittent episodes of muscular fasciculations, Lethal White Foal Syndrome (LWO) weakness, myotonia, or involuntary recumbency with varying intensity. Life-threatening complications could be caused by Material EB 1 ml, hair roots cardiac arrhythmias (secondary to the hyperkalaemia) as well Method TaqMan SNP assay as risk of suffocation by laryngospasm. HYPP is the result of a Breed Appaloosa, Paint Horse, Quarter Horse genetic mutation in the sodium channel gene of skeletal muscles. Inheritance Autosomal recessive It is inherited as an autosomal dominant trait. That means that a Duration 3 – 5 days heterozygous carrier of the defect gene shows the same symptoms Note The syndrome occurs in white foals born to American Paint Horses like a horse with both alleles affected. of overo lineage, specifically the frame overo subtype. Affected foals appear normal at birth but fail to pass meconium and develop Immune Mediated Myositis & MYH1 Myopathy (MYHM) severe colic as a result of ileus caused by a functional intestinal obstruction. In the absence of veterinary intervention, death ensues, Material EB 1 ml, hair roots usually within 24 to 48 hours postpartum. Method Sequencing Breed Appaloosa, Paint Horse, Quarter Horse Inheritance Autosomal dominant with variable penetrance Naked Foal Syndrome (NFS) Duration 1 – 2 weeks Material EB 1 ml, hair roots Note Quarter Horse and related breeds are susceptible to developing Method TaqMan SNP assay rapid onset of muscle atrophy and severe muscle damage at rest Breed Akhal-Teke (nonexertional rhabdomyolysis). An autoimmune muscle disease Inheritance Autosomal recessive called immune-mediated myositis (IMM) can cause this severe Duration 3 – 5 days atrophy, which can result in the loss of 40% of muscle mass within Note NFS is a genodermatosis in the Akhal-Teke horse breed. Affected 72 hours in Quarter Horse and related breeds. IMM is characterised horses have almost no hair and show a mild ichthyosis. They often by stiffness, weakness and nonspecific malaise. Affected horses are die within days to months after birth. However, the reason for these usually 8 years and younger or 17 years and older, environmental early deaths is not known, and some hairless foals have survived up factors combined with genetic susceptibility are important triggers to 2.5 years. for the development of muscle atrophy or severe rhabdomyolysis. Another clinical presentation of the MYH1 variant in young Quarter Ocular Squamous Cell Carcinoma (SCC) Horses is severe, sudden muscle damage not associated with exercise (nonexertional rhabdomyolysis). Horses with nonexertional Material EB 1 ml, hair roots rhabdomyolysis do not necessarily have muscle atrophy. Method Sequencing Breed Ardennes Horse, Belgian Horse, Haflinger 332 333 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Horse 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Horse LABORATORY FOR CLINICAL DIAGNOSTICS Inheritance Autosomal recessive Warmblood Fragile Foal Syndrome (WFFS) Duration 1 – 2 weeks Material EB 1 ml, hair roots Note Squamous cell carcinoma (SCC) is the second most common type Method TaqMan SNP assay of tumour in the horse and the most frequent tumour of the horse’s Breed Austrian Warmblood, Bavarian Warmblood, Belgian Warmblood, eye. Factors thought to increase risk for SCC include UV exposure, Brandenburger, Czech Warmblood, Danish Warmblood, Dutch pigmentation, and genetics. When originating at the limbus, SCC Warmblood, German Riding Pony, German Sport Horse, Hanoverian can spread into the cornea and quickly lead to visual impairment horse, Hessen, Holsteiner horse, Irish Sport Horse, Latvian horse, and destruction of the eye. This risk factor does not explain all cases Mecklenburger, Oldenburg horse, Rhinelander horse, Saxonian of ocular SCC but it appears to be a major contributor in Haflingers Warmblood, Selle Francais, Swedish Warmblood, Swiss Warmblood, and Belgians. Trakehner, Westphalian horse, Wielkopolski, Württemberger, Homozygous horses (R/R) are advised to have routine eye exams Zweibrücker, other/unknown performed by a veterinary ophthalmologist for early detection and Inheritance Autosomal recessive better prognosis, and to wear a UV protecting fly mask when out Duration 3 – 5 days during the daylight hours. Note Warmblood fragile foal syndrome (WFFS) is an inherited systemic connective tissue disorder that is prevalent in Warmblood horses. Polysaccharid Storage Myopathy Type 1 (PSSM) Symptoms are comparable to the Ehlers-Danlos-Syndrome in Material EB 1 ml, hair roots humans. The skin lacks tensile strength (extreme skin fragility Method TaqMan SNP assay characterised by tearing, ulceration, etc. from contact with normal Breed All surroundings). Inheritance Autosomal dominant Laboklin owns the exclusive license to perform this genetic test. Duration 3 – 5 days Note Horses with PSSM have signs typically associated with tying-up. Most commonly these signs are muscle stiffness, sweating and 20.4.2 Coat Colour/Coat Structure Horse reluctance to move. Episodes usually begin after very light exercise such as 10 – 20 minutes of walking and trotting. Agouti Laboklin owns the exclusive license to perform this genetic test. Material EB 1 ml, hair roots Method FLP Severe Combined Immuno Deficiency (SCID) Breed All Material EB 1 ml, hair roots Duration About 1 week Method FLP Breed Arabian Horse Appaloosa Pattern 1 Inheritance Autosomal recessive Duration 1 week Material EB 1 ml, hair roots Method Sequencing Note SCID of Arabian foals is a fatal disease caused by the lack of B- and Breed Appaloosa, Pony of the Americas, others T-lymphocytes. Thus, a foal affected by SCID is born with no immune Duration 1 – 2 weeks system, and generally dies of an opportunistic infection such as pneumonia, usually before the age of five or six months. Brindle 1 Material EB 1 ml, hair roots Method Sequencing Breed All Duration 1 – 2 weeks 334 335 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Horse 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Horse LABORATORY FOR CLINICAL DIAGNOSTICS Camarillo White - W4* Graying* Material EB 1 ml, hair roots Material EB 1 ml, hair roots Method Partner laboratory Method Partner laboratory Breed All Breed All Duration 3 – 4 weeks Duration 3 – 4 weeks Champagne Incontinentia Pigmenti (Hyperpigmentation) Material EB 1 ml, hair roots Material EB 1 ml, 20 – 30 hair roots from mane or tail Method TaqMan SNP assay Method Sequencing Breed All Breed All Duration 3 – 5 days Duration 1 – 2 weeks Note IP is an ectodermal dysplasia characterised by skin lesions evolving Chestnut over time, as well as dental, nail and ocular abnormalities. Affected Material EB 1 ml, hair roots horses also have streaks of darker and lighter coat coloration from Method TaqMan SNP assay birth. Due to X-linked dominant inheritance IP symptoms can only be Breed All seen in female individuals while affected males die during develop- Duration 3 – 5 days ment in utero. Cream Leopard Complex Material EB 1 ml, hair roots Material EB 1 ml, hair roots Method TaqMan SNP assay Method TaqMan SNP assay Breed All Breed All Duration 3 – 5 days Duration 3 – 5 days Dun Pearl* Material EB 1 ml, hair roots Material EB 1 ml, hair roots Method FLP Method Partner laboratory Breed All Breed On request Duration Approx. one week Duration 3 – 4 weeks GQ Santana Dominant White W10* Roan Zygosity* Material EB 1 ml, hair roots Material EB 1 ml, hair roots Method Partner laboratory Method Partner laboratory Breed All Breed On request Duration 3 – 4 weeks Duration 3 – 4 weeks 336 337 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Horse 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cattle LABORATORY FOR CLINICAL DIAGNOSTICS Sabino-1 20.5 Cattle Material EB 1 ml, hair roots Method Sequencing Sampling instructions Breed All There should not be any blood samples sent in for cattle from multiple births Duration 1 – 2 weeks because of a possible blood chimerism, but if the test allows it, hair roots, sperm or tissue samples can be used. One exception to this is the free martin test, for which a blood sample is mandatory. Silver Dapple Material EB 1 ml, hair roots Method Sequencing 20.5.1 Hereditary Diseases Breed All Duration 1 – 2 weeks Arachnomelia* (spider limbs) Material EB 3 – 5 ml, about 50 hair roots, tissue, sperm Splashed White Breed Fleckvieh Material EB 1 ml, hair roots Duration About 2 weeks Method Sequencing Note Hereditary arachnomelia of Fleckvieh cattle is passed on autosomal Breed All recessively. It is characterised by a developmental disorder of the Duration 1 – 2 weeks skeletal system leading to the birth of dead or malformed calves and an increased risk of injury to the mother. The test is not suitable for Tobiano arachnomellia of Brown Swiss, as both breeds have different gene Material EB 1 ml, hair roots mutations. Method FLP Breed All Bovine Leukocyte Adhesion Deficiency (BLAD) Duration 3 – 5 days Material EB 1 ml Breed Holstein-Friesian cattle Duration 1 week 20.4.3 Performance Horse Note BLAD is a fatal autosomal recessive disease of the immune system in Holstein cattle. Affected calves suffer from immunodeficiency and Speed-gene* die before reaching sexual maturity. Clinical signs manifest them- Material EB 1 ml, hair roots selves in recurrent non-specific infections of the respiratory system Method Partner laboratory and the gastrointestinal tract, delayed wound healing and reduced Breed Thoroughbred weight gain as well as leukocytosis with granulocytosis and lympho- Duration 1 – 2 weeks penia as laboratory findings. The test is based on the duplication and sequence analysis of the affected gene segment. Healthy homozygous animals, hetero zygous carriers and homozygous cattle suffering from the disease can be identified. 338 339 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cattle 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cattle LABORATORY FOR CLINICAL DIAGNOSTICS Bovine Progressive Degenerative Myeloencephalopathy* (Weaver Syndrome) Bovine Spinal Dysmyelination* (BSD) Material EB 3 – 5 ml, about 50 hair roots, tissue, sperm Material EB 3 – 5 ml, about 50 hair roots, tissue, sperm Breed Brown Swiss Breed Brown Swiss Duration Approx. 2 weeks Duration Approx. 2 weeks Note Weaver syndrome is a hereditary CNS disease in Brown Swiss. At Note BSD occurs in Brown Swiss, is autosomal recessive and leads the age of a few months, the first symptoms appear as weakness of to insufficient myelination. After birth, calves lie down in a lateral the hind legs, problems getting up and unsteady gait. Disorders are position with their limbs stretched forward. The head is often in a progressive and, after 1 – 3 years lead to recumbency and death. „moon-gazing position“; sometimes hyperreflexia can be observed. Weaver syndrome appears to be autosomal recessive, but genetics The animals are usually euthanised shortly after birth. is not yet fully understood. This indirect genetic test determines the probability of the disease. Muscular Atrophy* (SMA) Material EB 3 – 5 ml, about 50 hair roots, tissue, sperm Chondrodysplasia* Breed Brown Swiss Material EB 1 – 2 ml, about 30 hairs with roots, tissue, sperm Duration Approx. 2 weeks Breed Dexter, Dahomey Note This autosomal recessive disease in Brown Swiss calves leads to Duration Approx. 3 weeks the destruction of motor neurons and, at the age of a few weeks, to Note If, in Dexter cattle, both parents pass on the chondrodysplasia gene, spinal muscular atrophy. The animals are recumbent, usually without it leads to disproportionate dwarfism in calves (bulldog calves). The losing their appetite. Spinal reflexes are reduced, pumping respira- animals are generally not viable and are aborted or stillborn. tion is seen, and the animals often develop secondary pneumonia and die after a few weeks. Complex Vertebral Malformation* (CVM) Material EB 1 – 2 ml, about 30 hairs with roots, tissue, sperm Free Martins* Breed All Material EB 1 ml Duration 3 weeks Breed All Note CVM is an autosomal recessive hereditary disease of the (Holstein) Duration 2 – 3 weeks cattle that leads to disturbed bone and skeletal formation, reduced Note In case of twins of different sexes, about 90% of infertile, externally weight and, possibly, to changes in the heart. The foetuses are female calves occur during pregnancy, so-called free martins, due to resorbed or the calves are born dead. The disease occurs in many the transfer of male cells to the female embryo. Already in newborn breeding lines. twins of different sexes, the phenotypically female calves can be examined to see whether they develop as free martins. Factor XI Deficiency* Material EB 1 – 2 ml, about 30 hairs with roots, tissue, sperm Breed All Duration Approx. 5 weeks Note Factor XI deficiency leads to coagulopathy. Affected animals suffer from bleeding tendency and exhibit clinical symptoms caused by coagulopathy. The disease is autosomal recessive. 340 341 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Cattle 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics LABORATORY FOR CLINICAL DIAGNOSTICS 20.5.2 Breed Characteristics Cattle Kappa-Kasein* Material EB 3 – 5 ml, about 50 hair roots, tissue, sperm Breed All Double Muscling* Duration About 3 weeks Material EB 1 – 2 ml, about 30 hairs with roots, tissue, sperm Note The kappa-casein gene influences important parameters for milk Breed Belgian Blue, Piedmontese cattle processing. Kappa-casein variant B is particularly favourable for Duration 5 weeks further processing. Note Double muscling is a muscular hypertrophy due to an increase in volume of the muscle fibres. This genetic factor is mainly found in β-Lactoglobulin* cattle of the Piedmontese and Belgian Blue breeds. On the one Material EB 1 – 2 ml, about 30 hairs with roots, tissue, sperm hand, it leads to an increase in muscle mass by 20% and leaner Breed All meat, but on the other hand, it results in heavy births. Duration Approx. 5 weeks There are numerous other genes which cause muscle hypertrophy Note The A variant of ß-lactoglobulin is associated with increased milk in dominant or recessive inheritance and which can be detected, on yield. The B variant leads to a higher protein and casein content and request, in a further test in all cattle breeds. to an increased yield in the cheese dairy. Polledness* Red Factor* Material EB 3 – 5 ml, about 50 hair roots, tissue, sperm Material EB 3 – 5 ml, about 50 hair roots, tissue, sperm Breed All Breed Black pied Holstein Friesian cattle Duration Approx. 2 weeks Duration Approx. 2 weeks Note In polled cattle, the painful procedure of dehorning is no longer Note Black pied cattle of the breed Holstein Friesian with a predisposition required. Reservations regarding loss of performance in polled lines for a red coat („red factor“) are popular crossbreeds in red pied have largely been eliminated. However, the test cannot evaluate the breeding. The determination of the genotype of the MSHR gene genetic predisposition for scurs and has not been validated for zebu allows to differentiate between black-coloured cattle with red factor cattle. (Ee) and without red factor (EE). Milk Protein 20.6 Small Ruminants and New World Camelids The composition of milk proteins is of considerable importance for further milk processing. In particular, the cheese-making properties of the milk strongly depend on the milk composition. 90% of the proteins in milk consist of the six proteins αS1-casein, 20.6.1 Hereditary Diseases αS2-casein, β-casein, kappa-casein, α-lactalbumin and β-lactoglobulin. Arachnomelia* (spider lamb syndrome) -Kasein β Material 20 – 30 hairs with roots Material EB 1 – 2 ml, about 30 hairs with roots, tissue, sperm Species Sheep of all breeds Breed All Duration 3 – 4 weeks Duration 3 weeks Note Hereditary chondrodysplasia leads to underdeveloped muscles in Note The four most important genetic variants of -casein can be β the lamb and, at about 4 – 6 weeks of age, to skeletal deformities detected: A1, A2, A3 and B. Alternatively, the test can be performed of the head, spine, ribs and abnormally long and bent/twisted limbs for the two variants A1 and A2 only. (spider lambs). The autosomal recessive disease first occurred in black-headed sheep of the Suffolk and Hampshire breeds and is based on a mutation in the gene FGFR3 (Fibroblast Growth Factor Receptor 3). 342 343 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics 2019/20 Hereditary Diseases/Phenotype/Breed Characteristics - Pig LABORATORY FOR CLINICAL DIAGNOSTICS Microphthalmia* 20.6.2 Breed Characteristics Material EB 1 – 2 ml, swab, tissue, sperm Small Ruminants and New World Camelids Species Texel sheep Duration Approx. 5 weeks Milk Protein α-S1-Kasein* Note Microphthalmia leads to extremely small or missing eyeballs. Affec- Material 20 – 30 hairs with roots ted lambs are completely blind. In Texel sheep, microphthalmia is Species Goats of all breeds autosomal recessive. Duration 3 – 4 weeks Note The αS1-casein gene influences the casein and fat content of goat Predisposition for Scrapie* milk. A high content of αS1-casein is positive for cheese production, Material EB 1 – 2 ml, swab a low content is beneficial for people with milk intolerance. The gene Species Sheep variants A and B are associated with high αS1-casein contents, Duration Approx. 5 weeks while only little αS1-casein is formed in E, F and N. Note Scrapie is a transmissible prion disease in sheep and goats. Prions are proteins. If they are pathologically altered, they induce their own proliferation and the vacuolation of nerve cells, especially in the brain 20.7 Pig stem. Initially, affected animals are lazy, later they show increasing excitability and an unnatural gait and die within six months after the onset of the disease. In sheep, there is a genetic predisposition to Malignant Hyperthermia (MH)* the classical form of scrapie. Based on the amino acid pattern, five Material EB 1 ml genotype classes are distinguished. The risk of scrapie varies from Species Pig an extremely low (“resistant”) to a very high risk depending on the Duration 2 weeks genotype class. The EU has obliged its member states to breed Note Malignant hyperthermia syndrome or porcine stress syndrome (PSS) scrapie-resistant sheep. is passed on recessively and is mainly found in breeds with increa- sed muscle mass and reduced amount of fat. The disease is caused Free Martins* by a mutation of the ryanodine receptor in the skeletal muscle, Material EB 1 ml which leads to a disturbance of the Ca²-ion exchange and a lowered Species Sheep, goat, llama, alpaca threshold for muscle cell contraction. It is accompanied by hyper- Duration 3 – 4 weeks metabolism and increased body temperature caused by inhalation narcotics, muscle relaxants and stress. Damage to nerve, liver and Note In case of twins of different sexes, the transfer of male cells to the kidney tissue occurs. female embryo during pregnancy can result in infertile, externally female animals, so-called free martins. In llamas and alpacas, the risk is 90%, for sheep and goats < 1%, but increases with multiple pregnancies with four or more animals. Already in newborn twins of different sexes, the phenotypically female calves can be examined to see whether they develop as free martins. 344 345 2019/20 DNA Profile, Breed, Species 2019/20 DNA Profile, Breed, Species LABORATORY FOR CLINICAL DIAGNOSTICS 21 DNA Profile, Breed, Species Likelihood Ratio (relationship analysis) Material EB 1 ml, buccal swab Method Microsatellite analysis (STRs) + database analysis 21.1 Identity and Parentage Species Dog Duration 2 – 3 weeks The DNA profile of an animal is also called genetic fingerprint. In contrast to other Note If only one parent (e.g. the father) is available for a proof of marking methods, like microchips or tattoos, it cannot be manipulated or destroyed by parentage, this test allows to calculate a so-called probability value. external factors, such as injuries. It remains unchanged for a lifetime. On the one hand, Values can also be determined to evaluate full or half siblings if only a DNA profile enables a lifelong, doubtless identification of the animal. On the other samples of the potential siblings are available. hand, parentage (fatherhood or parenthood) can be proven with certainty by comparing What is important to know: The test is limited to the breeds in the genetic fingerprints of the family members. our database (current information can always be found on our homepage). Parentage (paternity test) Material EB 1 ml, buccal swab Horse and water buffalo: 20 – 30 hairs with roots 21.2 Breed and Species Method Microsatellite analysis (STRs) Species Dog, cat, horse, cattle, sheep, goat, llama, alpaca, water buffalo, pig Breed Analysis (database analysis) Duration 1 – 2 weeks Material EB 1 ml, buccal swab 4 – 5 weeks (alpaca, llama, water buffalo) Method Microsatellite analysis (STRs) + database analysis Note The proof of parentage (paternity test) makes it possible to check Species List Dog, cat which parents the offspring has. The DNA profiles of the parents and Duration 3 – 4 weeks offspring are the basis for this. Note The genetic classification of breeds allows for the statistical What is important to know: Even if only the paternity should be calculation of a classification probability of the breeds in our clarified, please send in samples from both parents. database. The test can be used to check the purity of breeds as well as to detect mixed breeds of the first generation. Thus, the test DNA Profile mainly allows to clarify the genetic proportions of so-called „list dog Material EB 1 ml, buccal swab breeds“ or to detect non-pure breeding. Among others, the DNA Horse and water buffalo: 20 – 30 hairs with roots profile of the animal serves as a basis for the test. Method Microsatellite analysis (STRs) (ISAG 2006) What is important to know: The test is limited to the breeds in Species Dog, cat, horse, cattle, sheep, goat, llama, alpaca, water buffalo, pig our database (current information can always be found on our Duration 1 – 2 weeks homepage). 4 – 5 weeks (alpaca, llama, water buffalo) Note To create a DNA profile, we test so-called microsatellite markers (e.g. 22 markers in dogs), which are recommended by the “Inter- national Society for Animal Genetics (ISAG)”. The DNA profiles we generate are internationally comparable with laboratories working according to the recommendations of ISAG. 346 347 2019/20 DNA Profile, Breed, Species 2019/20 Hygiene Examinations LABORATORY FOR CLINICAL DIAGNOSTICS Species Differentiation, Molecular Biological 22 Hygiene Examinations Material Miscellaneous (possibly consultation by telephone) Method Sequencing and database analysis In the veterinary practice, hygiene is one decisive prerequisite for successful treatment. Duration 2 – 3 weeks For this purpose, the proper functioning of sterilisers should be checked on a regular basis. The same holds true for testing the disinfection of surfaces or endoscopes. Our Note This method allows, for example, to assign certain signs to an research data reveals that several sterilisers we tested in practice did not show any animal species, because it is rarely possible to draw conclusions level of sterilisation anymore and particularly with endoscopes, regular monitoring of about the origin of samples such as faeces, trails of blood, etc. with disinfection measures is strongly recommended. the naked eye or with the help of common laboratory methods. In this test, a specific region of mitochondrial DNA is duplicated by PCR, sequenced and its origin analysed. Since this is a compara- tively sensitive method, even the smallest sample quantities can be 22.1 Profiles Hygiene assigned (e.g. blood spatter). The differentiation of animal species using the latest molecular bio- Hygiene Monitoring logical methods can be applied to a large number of questions, e.g. Monitoring of a dry heat steriliser OR monitoring of a steam steriliser + monitoring of “Does the neighbour‘s dog/cat do its business in our garden?” 3 surfaces (by contact plates) after disinfection. We would be pleased to inform you in advance by telephone in If you participate regularly (2x per year), you will get a certificate stating the successful order to clear up any confusion regarding the test. annual monitoring of the disinfection performance of your dry heat steriliser or steam steriliser and the surface disinfection test. Material Bioindicators + contact plates Method Culture Duration 7 days Note Request test set from the laboratory. 22.2 Single Tests Control of Cleaning and Disinfection of Surgical Instruments Material Bioindicators (screws and flexible tubes with blood and/or gravel and E. faecium) Method Culture Duration 3 – 7 days Note Request test set from the laboratory. Disinfectant Testing Material Disinhibitor broth Method Culture Duration 3 days Note Request disinhibitor broth from the laboratory. 348 349 2019/20 Hygiene Examinations 2019/20 Hygiene Examinations LABORATORY FOR CLINICAL DIAGNOSTICS Endoscope Control Hand and Surface Disinfection Test Material Rinse samples + contact plates Material Contact plates Method Culture Method Culture Duration 3 – 5 days Duration 2 days Note Request the contact plates and sterile test containers from the Note Request contact plates from the laboratory. laboratory in due time. Where applicable, the following multiresistant germs can be iden- It is required to return the collected samples to the laboratory within tified: MRSA (methicillin-resistant Staphylococcus aureus) and/or 24 hours. MRSE (methicillin-resistant Staphylococcus epidermidis) and ESBL (germs forming extended spectrum ß-lactamase). For this, additional Heat Steriliser Control costs will be incurred. If you participate regularly (2x per year), you will get a certificate Material Bioindicators (contaminated with Bacillus atrophaeus) stating the annual monitoring of the disinfection performance of your Method Culture surface disinfection test. Duration 7 days Note If required, the bioindicators should be requested from the Testing of Air Settlement Plates laboratory in due time. If you participate regularly (2x per year), you will get a certificate Material Air settlement plate stating the annual monitoring of the disinfection performance of your Method Culture heat steriliser. Duration 2 days Note Request air settlement plates from the laboratory. Steam Steriliser Control (autoclave) This test is not suitable for examinations of barns and stables! Material Bioindicators (contaminated with Bacillus atrophaeus and Geobacillus stearothermophilus) Method Culture Duration 7 days Note If required, the bioindicators should be requested from the laborato- ry in due time. If you participate regularly (2x per year), you will get a certificate stating the annual monitoring of the disinfection performance of your autoclave. 350 351 2019/20 Reference Ranges 2019/20 Reference Ranges LABORATORY FOR CLINICAL DIAGNOSTICS 23 Reference Ranges 23.1.2 Haematological reference ranges dog, cat, horse unit dog cat horse 23.1 Dog, cat, horse Anisocytosis neg. neg. neg. 23.1.1 Clinical chemistry Basophils % 0 0 – 1 0 – 2 unit dog cat horse Eosinophils % 0 – 6 0 – 6 0 – 4 Enzymes 25°C ALT U/l up to 55 up to 70 - Erythrocytes T/l 5.5 – 8.5 5.0 – 10.0 6.0 – 12.0 α-Amylase U/l up to 1650 up to 1850 up to 170 AP U/l up to 108 up to 140 up to 450 Haematocrit l/l 0.44 – 0.52 0.3 – 0.44 0.3 – 0.5 AST (GOT) U/l up to 25 up to 30 up to 250 Haemoglobin g/l 150 – 190 90 – 150 110 – 170 Cholinesterase U/l 1500 – 3000 1000 – 3000 1500 – 3000 CK U/l up to 90 up to 130 up to 130 (190) Hypochromasia neg. neg. neg. GLDH U/l up to 6 up to 6 up to 8 γ-GT U/l up to 5 up to 5 up to 25 Leukocytes G/l 6 – 12 6 – 11 5 – 10 α -HBDH U/l up to 50 up to 97 up to 170 % 13 – 30 15 - 38 20 – 45 LDH U/l up to 100 up to 70 up to 400 Lymphocytes Lipase (DGGR) U/l up to 120 up to 26 up to 250 Monocytes % 0 – 4 0 – 4 0 – 5 Substrates Albumin g/l 25 – 44 26 – 56 25 – 54 Segmented % 55 - 75 60 – 78 45 – 70 Albumin-Globulin > 0.59 > 0.6 0.7 – 1.1 (A/G) -Quotient Unsegmented % 0 – 4 0 – 4 0 – 6 Bile Acids µmol/l up to 20, up to 20, up to 12 post-prandial up to 40 post-prandial up to 40 Platelets G/l 150 – 500 180 – 550 90 – 300 Bilirubin I (total) μmol/l up to 3.4 up to 3.4 8.6 – 59.9 Cholesterol mmol/l 3.1 – 10.1 1.8 – 3.9 1.81 – 4.66 Differential blood count (absolute numbers) Creatinine μmol/l 35 – 106 up to 168 71 – 159 Fructosamine μmol/l up to 374 up to 340 up to 360 Segmented G/l 3 – 9 3 – 11 3 – 7 Globuline g/l < 45 < 55 24 - 51 Lymphocytes G/l 1 – 3.6 1 – 4 1.5 – 4 Glucose mmol/l 3.05 – 6.1 3.1 – 6.9 3.05 – 4.99 Lactate mmol/l 0.5 – 3.0 up to 1.0 0.5 – 2.0 Monocytes G/l 0.04 – 0.5 0.04 – 0.5 0.04 – 0.4 NEFA mmol/l 0.1 – 0.5 0.1 – 0.5 0.1 – 0.5 Protein (total) g/l 54 – 75 57 – 94 55 – 75 Eosinophils G/l 0.04 – 0.6 0.04 – 0.6 0.04 – 0.3 Triglycerides mmol/l up to 3.9 up to 1.14 up to 0.97 Basophils G/l up to 0.04 up to 0.04 0 – 0.15 Urea mmol/l 3.3 – 8.3 5.0 – 11.3 3.3 – 6.7 β -HBS mmol/l up to 0.6 up to 0.6 up to 0.6 Unsegmented G/l up to 0.5 up to 0.6 0 – 0.6 Minerals and Trace Minerals Reticulocytes /nl < 110 < 60 - Calcium mmol/l 2.3 – 3.0 2.3 – 3.0 2.5 – 3.4 Chloride mmol/l 96 – 113 110 – 130 95 – 105 Copper μmol/l 15.7 – 18.9 13.4 – 16.9 7.9 – 21.0 Iron μmol/l 15 – 45 8 – 31 17.9 – 64.5 Potassium mmol/l 3.5 – 5.1 3.0 – 4.8 2.8 – 4,5 Magnesium mmol/l 0.6 – 1.3 0.6 – 1.3 0.5 – 0.9 Sodium mmol/l 140 – 155 145 – 158 125 – 150 Phosphate mmol/l 0.7 – 1.6 0.8 – 1.9 0.7 – 1.5 Selenium μg/l 80 – 250 80 – 250 100 – 200 Zinc μmol/l 7.7 – 19.9 12.2 – 15.3 9.2 – 19.9 Selenium horse: Up to 70 µg/l are marginal, more than 300 µg/l high/critical. Foals and Iceland horses are sometimes well below these levels. 352 353 2019/20 Reference Ranges 2019/20 Reference Ranges LABORATORY FOR CLINICAL DIAGNOSTICS 23.1.3 Hormones dog, cat, horse 23.2 Reference ranges rabbit, guinea pig and ferret 23.2.1 Clinical chemistry unit dog cat horse mid-Nov. – mid-July up to 30 unit rabbit guinea pig ferret ACTH pg/ml 6 – 58 up to 110 mid-July – mid-Nov. < 50 Enzymes 25°C m-neutered: < 0.1 granulosa theca ALT (GPT) U/l up to 61 up to 61 55 – 206 Anti- m-intact: > 2.0 w-neutered: < 0.1 cell tumour: over 4 Müllerian ng/ml w-neutered: < 0.02 w-intact: > 2.0 male neutered: ≤ 0.1 α-Amylase U/l up to 459 up to 3159 21 – 59 Hormone w-intact: > 0.5 male intact: > 2 AP U/l 9.05 – 94.58 up to 418 18 – 71 Cortisol ng/ml 5 – 65 3 – 50 (130) 30 - 67 AST (GOT) U/l 3.75 – 32.44 up to 90 43 – 142 Insulin ng/ml 8 – 25 10 – 30 up to 23.4*** CK U/l 1.63 – 559.53 up to 2143 80 – 453 Oestradiol pg/ml prooestrus: 25 – 65 interoestrus: up to 20 prooestrus: 1.2 – 6.2 oestrus: up to 25 oestrus: 20 – 60 oestrus: 7.1 – 13.0 GLDH U/l 0.68 – 14.78 up to 17 up to 2 anoestrus: up to 30 - dioestrus: 3.7 – 5.0 neutered: up to 10 - - γ-GT U/l 2.5 – 14.46 up to 13 up to 10 males: up to 15 - - Sertoli cell tumour: - - Lipase U/l up to 1587 up to 152 86 - 334 over 30 Substrates Progesterone ng/ml prooestrus: up to 1.0 Preov: up to 1.0 Levels over 1.0 indicate oestrus: up to 30 Postov: over 1.0 luteal activity Albumin g/l 36 – 57 - 26 – 42 ovulation*: 4.0 – 8.0 - - anoestrus: up to 1.0 - - Cholesterol mmol/l 0.3 – 1.7 0.3 – 1.7 2.4 – 6.9 - Creatinine μmol/l 51.38 – 154.35 up to 77 21 – 69 Fructosamines μmol/l 248.08 – 501.43 up to 271 128 – 201 Testosterone ng/ml m: 1.5 – 8.5 m: 2.5 – 7.0 stallion: 1.0 – 5.0 w: up to 0.4 - gelding: < 0.04** Glucose mmol/l 5.8 – 14.8 5.0 – 16.0 2.7 – 8.6 m-neutered: up to 0.5 m-neutered: up to 0.5 mare: < 0.04** Protein (total) g/l 48.66 – 73.64 44 – 66 54 – 78 TSH ng/ml up to 0.6 - - Triglycerides mmol/l 0.5 – 3.4 0.3 – 2.4 0.5 – 1.9 TSH μU/ml - > 0.04 - Urea mmol/l 2.63 – 10.28 3.3 – 10.3 5.1 – 16.6 T3 ng/dl 20 – 206 33 – 167 25 – 180 Minerals and f T3 pmol/l 3.7 – 9.2 0.8 – 1.4 1.1 – 7.2 Trace Minerals T4 μg/dl 1.3 – 4.5 0.9 – 2.9 1.3 – 4.1 Calcium mmol/l 3.02 – 4.3 2.4 – 3.1 2.0 – 2.6 T4 pmol/l 7.7 – 47.6 6.4 – 33.3 9.0 – 44.9 f Iron μmol/l 20 – 59 26 – 76 16 – 55 * Time of mating for female dogs: Optimum 24 to 48 hours, maximum 96 hours after ovulation Potassium mmol/l 3.52 – 6.04 4.5 – 8.8 3.8 – 5.5 ** Testosterone gelding: No increase in levels to the standard range after HCG stimulation. Magnesium mmol/l 0.66 – 1.51 1.0 – 2.6 0.9 – 1.6 Testosterone mare: Increased levels indicate a granulosa theca cell tumour. *** The reference value was test-specifically adapted according to the latest literature. Sodium mmol/l 132.61 – 154.0 130 – 150 139 – 166 Phosphate mmol/l 0.54 – 2.18 1.0 – 7.0 1.0 – 3.0 Hormones Androstenedione ng/ml - - < 428 17-OH- Progesterone ng/ml - - < 26.1 Up to 0.5: neutered 0.5 – 1: male: 1.5 – 8.5 questionable male/neutered: female: up to 0.4 Testosterone ng/ml over 1: testicular up to 0.5 neutered: up to tissue probably 0.5 present T4 μg/dl 3.9 – 5.3 1.1 – 5.2 1.1 – 2.8 fT4 pmol/l up to 20 (30) up to 20 (30) - 354 355 2019/20 Reference Ranges 2019/20 Reference Ranges LABORATORY FOR CLINICAL DIAGNOSTICS 23.2.2 Haematological reference ranges rabbit, guinea pig, ferret 23.3 Reference ranges birds unit rabbit guinea pig ferret 23.3.1 Clinical chemistry Erythrocytes T/l 4.37 – 7.43 4.51 – 6.36 7.4 – 13.0 unit parakeets amazon parrots parrot Enzymes 25°C l/l 0.28 – 0.48 0.39 – 0.55 0.4 – 0.7 Haematocrit ALT (GPT) U/l 5 – 20 5 – 11 5 – 12 Haemoglobin g/l 89.63 – 153.82 117 – 169 138 – 209 α-Amylase U/l 187 – 582 100 – 600 200 – 600 AP U/l 10 – 326 15 – 311 20 – 311 Leukocytes G/l 2.71 – 12.23 2.9 – 14.4 3.0 – 16.7 AST (GOT) U/l 55 – 390 35 – 350 100 – 400 Cholinesterase U/l 2000 – 4000 2000 – 4000 2500 – 12000 Segmented % 32 – 64 12 – 62 19 – 79 CK U/l 54 – 300 100 – 500 130 – 400 Lymphocytes % 13 – 54 28 – 84 16 – 75 GLDH U/l 0 – 9.9 0 – 9.9 0 – 9.9 γ-GT U/l 1 – 30 1 – 12 1 – 30 Monocytes % 3 – 14 0 – 9 0 – 7 LDH U/l 76 – 450 150 – 400 150 – 400 Substrates Eosinophils % < 3 0 – 14 0 – 6 Albumin g/l 7 – 18 19 – 35 15 – 33 mmol/l 3 – 6 4 – 6 2.6 – 7 Basophils % < 9 0 – 2 0 – 2 Cholesterol Protein g/l 22 - 50 26 – 50 26 – 53 Unsegmented % 0 0 – 1 0 – 2 Bile acids μmol/l up to 100 20 – 150 19 – 144 Uric acid μmol/l 190 – 833 70 – 595 100 – 654 Hypochromasia neg. neg. neg. Urea mmol/l - 0.9 – 9.6 0.7 – 9.5 Anisocytosis neg. neg. neg. Triglycerides mmol/l 1.23 – 3.05 0.66 – 2.25 0.57 – 1.58 Minerals and Trace Minerals Platelets G/l 225.45 – 905.3 273 – 745 297 – 910 Calcium mmol/l 1.6 – 3.3 2.0 – 3.5 1.75 – 3.5 Differential blood count (absolute numbers) Potassium mmol/l 2.2 – 4.6 2.0 – 4.5 2.0 – 5.0 Sodium mmol/l 139 – 159 125 – 160 135 – 165 Segmented G/l 0.87 – 7.82 0.9 – 5.1 3.2 – 13.1 Phosphate mmol/l 0.8 – 2.5 0.3 – 1.8 0.35 – 4.0 Lymphocytes G/l 0.36 – 6.58 1.4 – 10.7 2.6 – 12.5 23.3.2 Haematological reference ranges birds Monocytes G/l 0.08 – 1.71 up to 0.7 up to 1.1 unit parakeet amazon parrots parrot Eosinophils G/l 0.07 – 0.19 up to 1.5 up to 1.0 Haematocrit l/l 0.48 – 0.58 0.44 – 0.55 0.43 – 0.51 Haemoglobin g/l 124 – 175 135 – 200 142 – 171 Basophils G/l 0.06 – 1.1 up to 0.11 up to 0.3 Leukocytes G/l 3 – 11 1.5 – 15 6 – 15 Unsegmented G/l 0 up to 0.07 up to 0.3 Heterophils % 43 – 75 43 – 75 45 – 76 Lymphocytes % 20 – 58 20 – 55 20 – 51 Monocytes % 0 – 3 0 – 2 0 – 1 Eosinophils % 0 – 2 0 – 1 0 – 1 Basophils % 0 – 1 0 – 2 0 – 1 Unsegmented % - - - Hypochromasia neg. neg. neg. Anisocytosis neg. neg. neg. Platelets G/l 34.4 – 38.4 - 46 – 49 356 357 2019/20 Reference Ranges 2019/20 Reference Ranges LABORATORY FOR CLINICAL DIAGNOSTICS 23.4 Reference ranges farm animals unit cattle sheep goat pig alpaca llama Vitamins 23.4.1 Clinical chemistry β -carotene μg/l over 2500 - - - - - Vitamin A μg/l 130 - 380 - - - - - unit cattle sheep goat pig alpaca llama Vitamin B12 pg/ml over 100 - - - - - Enzymes 25°C Vitamin D3 nmol/l 75 – 125 - - - - - ALT (GPT) U/l up to 50 up to 14 up to 14 up to 68 up to 50 up to 26 Vitamin E mg/l over 3 - - - - - α -Amylase U/l up to 161 up to 120 - up to 3500 - - Hormones AP U/l up to 300 45 – 235 45 – 235 up to 170 32 – 167 46 – 119 Insulin μU/ml up to 5 - - - - - AST (GOT) U/l up to 80 up to 60 10 – 50 up to 35 up to 308 up to 450 Progesterone ng/ml over 1.0** - - - - - Cholinesterase U/l 50 – 160 - - - - 50 – 100 T4 μg/dl 3.4 – 8.2 - - - - - CK U/l up to 250 10 – 50 up to 65 up to 500 up to 120 up to 137 Other values - GLDH U/l up to 8 up to 2 up to 7 up to 4 up to 19 up to 25 Haptoglobin g/l up to 0.35 - - up to 0.68 - - γ-GT U/l up to 20 up to 32 10 – 20 up to 45 up to 35 up to 28 IgG mg/dl 1700 – 2700* - - 1700 – 2900 - - GPX U/g Hb over 130 60 – 180 - - - - α -HBDH U/l up to 700 up to 700 - up to 300 - - * cattle (calf: > 800) LDH U/l up to 1500 up to 1500 up to 1500 up to 600 up to 433 up to 695 ** values that exceed 1.0 indicate luteal activity Lipase U/l 2 – 8 - - - - - Substrates 23.4.2 Haematological reference ranges farm animals Albumin g/l 30 – 40 24 – 30 30 – 40 18 – 31 29 – 43 29 – 50 Bilirubin I (total) μmol/l up to 5.0 up to 8.5 up to 8.5 up to 4.3 up to 6.8 up to 8.6 Cholesterol mmol/l 2.07 – 3.88 1.2 – 1.9 2.07 – 3.88 2.0 – 3.3 0.4 – 2.3 0.3 – 2.3 unit cattle sheep goat pig alpaca llama Protein g/l 60 – 80 50 – 70 60 – 80 55 – 86 57 – 72 47 – 73 Erythrocytes T/l 5.0 – 10.0 7.3 – 11.3 8 – 18 5.8 – 8.1 9.4 – 18.1 9.9 – 17.7 Bile acids μmol/l 10 – 25 up to 10 - - - - Haematocrit l/l 0.28 – 0.38 0.29 – 0.38 0.24 – 0.48 0.33 – 0.45 0.22 – 0.45 0.25 – 0.46 Globulines g/l 27 – 48 27 – 48 27 – 48 51 – 64 21 – 31 12 – 32 Haemoglobin g/l 90 – 140 80 – 120 80 – 120 108 – 148 102 – 193 115 – 195 Glucose mmol/l 1.94 – 3.05 2.2 – 5.2 2.2 – 5.2 3.9 – 6.4 5.7 – 8.3 5.7 – 7.0 Leukocytes G/l 4 – 10 4 – 10 4 – 13 10 – 22 7.1 – 18.6 8.9 – 22.4 Urea mmol/l up to 8 4.5 – 10.7 4.5 – 10.7 3.3 – 8.3 3.6 – 10.1 3.2 – 12.8 Segmented % 25 – 45 10 – 50 30 – 48 10 – 39 49 – 65 49 – 65 Lymphocytes % 45 – 65 40 – 80 50 – 70 49 – 85 21 – 25 21 – 25 β -HBS mmol/l 0.2 – 1.0 up to 0.6 up to 0.6 - - - Creatinine μmol/l 88 – 177 50 – 120 50 – 120 40 – 130 88.4 – 212.2 79.5 – 247.5 Monocytes % 2 – 6 0 – 15 0 – 4 2 – 4 0 – 5 0 – 5 Lactate mmol/l 0.5 – 3.0 1 – 1.4 1 – 1.4 - - - Eosinophils % 1 – 10 0 – 8 1 – 8 0 – 6 6 – 22 6 – 22 NEFA mmol/l 0.4 – 0.8 0.1 – 0.5 0.1 – 0.5 - - - Basophils % 0 – 2 0 – 4 0 – 1 0 – 5 0 – 1 0 – 1 Triglycerides mmol/l 0.17 – 0.51 0.06 – 0.34 0.17 – 0.51 up to 0.5 up to 0.6 up to 0.27 Unsegmented % 0 – 3 0 – 4 0 – 2 0 – 7 0 – 1 0 – 1 Platelets G/l 300 – 800 200 – 800 200 – 800 175 – 580 200 – 600 200 – 600 Minerals and Trace Minerals Reticulocytes /ml up to 0.1 up to 0.1 up to 0.1 - - - Calcium mmol/l 2.3 – 2.8 2.1 – 2.7 2.2 – 2.8 2.4 – 3.5 2.1 – 2.5 1.9 – 2.7 Chloride mmol/l 90 – 110 75 – 114 97 – 110 102 – 106 99 – 122 103 – 122 Iron μmol/l 20 – 40 20 – 30 16 – 35 16.7 – 35.5 18.8 – 37.4 18.6 – 30.8 Potassium mmol/l 3.5 – 4.5 3.5 – 4.5 4.5 – 6.5 4.0 – 5.0 4.0 – 5.7 3.6 – 6.2 Cobalt μg/l 1.0 – 3.5 - - - - - Copper μmol/l 8 – 24 7 – 24 16 – 32 16 – 39 2.1 – 12.5 6.1 – 7.9 Magnesium mmol/l 0.8 – 1.3 0.8 – 1.0 0.8 – 1.0 1.1 – 1.5 0.7 – 1.0 0.8 – 1.1 Sodium mmol/l 135 – 145 145 – 155 135 – 157 140 – 160 146 – 155 148 – 158 Phosphate mmol/l 1.1 – 2.4 1.1 – 2.5 1.61 – 2.26 2.1 – 3.3 1.1 – 2.5 1.5 – 3.6 Selenium μg/l 40 – 85 55 – 170 15 – 40 100 – 200 over 99 over 99 Zinc μmol/l 8 - 24 11.0 – 20.5 10.7 – 19.9 10 – 20 3.0 – 14.6 4.1 – 12.4 358 359 2019/20 Conversion Table for Laboratory Diagnostic Parameters 2019/20 Conversion Table for Laboratory Diagnostic Parameters LABORATORY FOR CLINICAL DIAGNOSTICS 24 Conversion Table Potassium mg/dl 0.2557 mmol/l 3.9102 Copper µg/dl 0.1574 µmol/l 6.3532 for Laboratory Diagnostic Parameters Magnesium mg/dl 0.4113 mmol/l 2.4312 Sodium mg/dl 0.4350 mmol/l 2.2989 Phosphate mg/dl 0.3229 mmol/l 3.0974 On the diagnostic findings compiled by us, you will find the measured values as well as Zinc µg/dl 0.1530 µmol/l 6.5370 information on the standard ranges in the internationally valid SI units. During follow- up checks, you may want to compare the measured values of different findings using identical units of measurement. The conversion factors for the parameters for which we have changed the unit of measurement are listed below. 24.2 Blood Parameters To convert from one unit of measurement to the other, the corresponding measured Conversion factor Conversion factor Old unit to SI-unit SI-unit to old unit value must be multiplied with the conversion factor (e.g. bilirubin in mg/dl x 17.104 = bilirubin in μmol/l). Erythrocytes Mio/µl 1 T/l 1 Haematocrit % 0.01 l/l 100 Haemoglobin g/dl 10 g/l 0.1 24.1 Clinical-chemical Parameters Leukocytes 1/µl 0.001 G/l (= 109/l) 1000 Thrombocytes 1/µl 0.001 G/l (= 109/l) 1000 Reticulocytes % 0.001 1 1000 Conversion factor Conversion factor Old unit to SI-unit SI-unit to old unit Substrates You will find a converter for easily comparing diagnostic findings with parameters Albumin g/dl 144.9 µmol/l 0.0069 in different units – our SI calculator – in the menu item Vetinfo on our website Bilirubin mg/dl 17.104 µmol/l 0.0585 www.laboklin.com. Cholesterol g/dl 0.0259 mmol/l 38.664 Protein g/dl 10 g/l 0.1 Fibrinogen mg/dl 0.01 g/l 100 Glucose mg/dl 0.0555 mmol/l 18.016 Uric acid mg/dl 59.48 µmol/l 0.0168 Urea mg/dl 0.1665 mmol/l 6.0060 Creatinine mg/dl 88.402 µmol/l 0.0113 Lactate mg/dl 0.111 mmol/l 9.0080 Triglycerides mg/dl 0.0114 mmol/l 87.500 Minerals and Trace Minerals Calcium mg/dl 0.2495 mmol/l 4.0080 Chloride mg/dl 0.2821 mmol/l 3.5453 Iron µg/dl 0.1791 µmol/l 5.5847 360 361 2019/20 Courier Service/Invoicing 25 Courier Service LABOKLIN offers courier services in most countries. The samples are generally delivered to LABOKLIN within 24/48 hours. For more information, including prices and the possibilities of sample collection in your area, please contact our Service Department or your local LABOKLIN office. Our contacts: see page 8 and following. 26 Invoicing All prices listed on the submission forms are quoted without the applicable Value Added Tax (VAT). To receive VAT-free invoices, please provide your international tax number (EU only). We issue invoices at the beginning of the next month with detailed information on costs per sample and tests performed in the previous month, together with animal and owner name. If an invoice is to be sent to the owner, we invoice with a factor 1.4 plus 19% German VAT. This is only possible for genetic tests and when the owners´ signature and complete data are supplied. There are discounts available to veterinarians depending on the monthly invoice revenue: For more information please contact us or your local LABOKLIN office. 362 www.laboklin.com 97688 Bad Kissingen • Steubenstraße 4 D Tel. +49 (0) 971/7 20 20 • Fax +49 (0) 971/6 85 46 e-mail: [email protected] • www.laboklin.com