Diagnosis of bovine tuberculosis in Mozambique Comparison of diagnostic methods

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1 Diagnosis of bovine tuberculosis in Mozambique Comparison of diagnostic methods 1 Maxhuza, G., 2 Adelina Machado, A., 3 Manfred Tanner, 4 Irmgard Moser 1 National Department of Veterinary Services, Ministry of Agriculture, Maputo, Mozambique 2 Veterinary Faculty, Eduardo Mondlane University, Maputo, Mozambique 3 Institute of Infectious diseases and Zoonoses, Ludwig-Maximilian-University, Munich, Germany 4 Friedrich-Loeffler-Institut, Institute of Molecular Pathogenesis, Jena, Germany VI International M.bovis Conference 16th - 19th June 2014 Cathays Park, Cardiff, Wales, CF10 3ND, Great Britain

2 Tuberculosis (btb) prevalence in cattle Morrumbala 12% btb Govuro 60% btb Massingir 1% btb

3 Manifestation of tuberculosis and immune response in cattle Cellular Immunity Humoral Immunity Tuberkulin-Test Immune Response γ-ifn Anergy Time Bacterial Load Symptoms Pathology Non Visible Lesions Visible Lesions Modified according to de la Rua-Domenech et al., 2006

4 Animals and testing methods A Positive herd 106 animals 69/89 SICCT positive B Positive herd 80 animals 28 SICCT positive C Negative herd 100 animals 9 SICCT positive Single interdermal cervical comparative tuberculin test (SICCT) Gamma interferon release assay (IFN γ; Bovigam ) ELISA (IDEXX M. bovis Ab Test; IDEXX ) Meat inspection Bacteriological culture Molecular identification of isolates (PCR)

5 IFNƴ SICTT Culture Inspection IDEXX SICTT IFNƴ SICTT Inspection SICTT IDEXX Inspection IDEXX Synopsis of the results of diagnostic procedures in herd A 106 animals tested 1 IFNγ 2 SICCT 3 Culture 4 Inspection 5 IDEXX positive inconclusive negative not tested

6 BTB Detection methods Comparison of results Animals positive / Animals tested Method IFN γ SICCT Meat inspection Culture IDEXX N 28/103 14/103? 69/89 88/94 31/41 29/75 % /75? 6.6 SICCT IFNγ N % SICCT Meat N % insp.. SICCT IDEXX N % Meat IDEXX N % insp ? Total ? Total ? ? 2 3.2? Total ? Total

7 IFNγ and IDEXX results interpretation A/P>0.3 Positive A/P<0.3 Negative Positive OD PPDbov NIL antigen > 0.1 and OD PPDbov PPDav > 0.1 A : Sample P: Positive Control Negative OD PPDbov NIL antigen < 0.1 or OD PPDbov PPDav < 0.1 Inconclusive OD PPDbov NIL antigen > 0.1 and OD PPDbov PPDav > 0.05 Active cells OD PWM NIL antigen > 0.5 Active cells OD PWM NIL antigen > 0.3 OD values of positive samples 1, , , OD values of negative samples -0, , ,02907 OD values of inconclusive samples 0, ,20515

8 Meat inspection Routine Additionally for research Score of lesions (herd A) Masseter muscle Kidney Tongue Lymph nodes Lungs Tonsils Liver retropharyngeal Spleen parotidic Lymph nodes Prescapular Mediastinal Bronchial Precrural Mesenterial Inspection of the inner and outer wall of the carcass 53 Bronchial Ln. 49 Mesenterial Ln. 47 Mediastinal Ln. 34 Retropharyngeal Ln. 34 Lungs 30 Liver 13 Tonsils 13 Prescapular Ln. 10 Submandibular Ln. 9 Spleen 6 Mandibular Ln. 5 Parotidic Ln. 4 Diaphragma 3 Precrural Ln. 2 Kidney 2 Intercostal Ln. 1 Pleura/Percard

9 Pathological lesion at meat inspection

10 Summary and conclusion Four diagnostic methods to detect btb in a highly infected cattle herd have been compared: Single interdermal cervical comparative tuberculin test (SICCT) Gamma interferon release assay (IFN γ; Bovigam ) ELISA (IDEXX M. bovis Ab Test; IDEXX ) Meat inspection SICCT and meat inspection yielded the highest detection rate with 77.5% and 93.6%, respectively, and the highest single animal agreement with 77.4%. IGRA yielded the lowest detection rate with only 27.2%, 40.8% with severe interpretation. IDEXX ELISA yielded a detection rate of 38.6%, 45.2% with severe interpretation. Bacteriological culture yielded a detection rate of 75.6%. However, the species identification of the isolates has not yet been done. From the diagnostic methods for intra vitam diagnosis of btb SICCT yielded by far the highest detection rate.

11 Tuberculin skin testing rough conditions

12 Tuberculin skin testing using a mobile crush barrier

13 Many helpful hands available

14 Tables for veterinary equipment and documentation

15 Tuberculin skin testing normal procedure

16 Tuberculin skin testing normal procedure (cont.)

17 Tuberculin skin testing rough conditions

18 Tuberculin skin testing rough conditions (cont.)

19 Capturing escaped cows for testing

20 Tuberculin skin testing - Measuring skin fold thickness

21 Tuberculin skin testing - Injecting PPDs and harvesting blood

22 Positive skin test reactions

23 Accommodation in the field Lab Appartments back home from the field

24 Friendly contacts at the water supply

25 Lab work in the field and in Maputo

26 On the way to Malawi having negotiations Walking towards Rio Chire Crossing the border Border control

27 Fans and audients everywhere

28 Bem Vindo Mocambique! Obrigada pela vossa atenção! Thank you for your attention!

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