Fabrizio Agnoletti. Istituto Zooprofilattico Sperimentale delle Venezie.

Transcription

1 Fabrizio Agnoletti Istituto Zooprofilattico Sperimentale delle Venezie Mission: prevention, control and research activities in 3 main areas: animal health and welfare food safety environmental protection 1

2 Istituto Zooprofilattico Sperimentale delle Venezie High density of livestock farms High density of rabbit farms 4 % of the overall italian production of rabbit is concentrated in the Veneto region 2

3 ISTITUTO ZOOPROFILATTICO SPERIMENTALE DELLE VENEZIE High density of livestock farms High density of rabbit farms Development of considerable expertise on the diagnosis of the pathologies encountered in intensive rabbit farms Remarkable research activity Our customers: farmers, technicians, veterinarians, feed and pharmaceutical companies, public veterinary services etc. 3

4 Diagnosis of rabbit diseases Bacterial diseases Viral diseases Parasitic diseases Real time PCR for MV and RHDV 4

5 Diagnosis of rabbit diseases Bacterial diseases Viral diseases Enteric pathogens Skin/reproductive pathogens Respiratory pathogens Escherichia coli Klebsiella pneumoniae Salmonella sp. Clostridium sp. Parasitic diseases 5

6 Diagnosis of rabbit diseases Bacterial diseases Viral diseases Enteric pathogens Skin/reproductive pathogens Respiratory pathogens Staphylococcus aureus Pasteurella multocida Parasitic diseases 6

7 From isolation to further characterization of enteric (bacterical) pathogens E. coli BIOTYPING Sorbose Dulcitol D-Raffinose Sucrose Ramnosio isolation SEROTYPING (somatic antigens) A panel of 23 antisera GENOTYPING (detection of genes encoding E. coli virulence factors) eae (intimin) AF/R1, AF/R2 (adhesive factor/rabbit 1 and 2) 7

8 From isolation to further characterization of enteric (bacterical) pathogens Clostridium spiroforme Clostridium perfringens Clostridium difficile Bacterioscopic examination isolation isolation isolation PCR: identification PCR: binary toxin Toxinotyping multiplex PCR: detection of α, β, ε and ι toxins encoding genes duplex PCR: detection of β2 and enterotoxin encoding genes duplex PCR: β2 atypical/consensus Biochemical identification multiplex PCR: identification and detection of toxins A and B encoding genes multiplex PCR: detection of binary toxin encoding genes 8

9 From isolation to further characterization of skin/reproductive pathogens Staphylococcus aureus Beta-haemolysin isolation BIOTYPING Staphylokinase Coagulation of bovine plasma Crystal-violet reaction IDENTIFICATION (PCR) DETECTION OF METHICILLIN RESISTANT STRAINS (meca gene) 9

10 ANTIMICROBIAL SUSCEPTIBILITY TESTING (for therapy, research, cooperation with pharmaceutical companies) 1. Disk diffusion test (Kirby Bauer method) 2. MIC (Minimum Inhibitory Concentration) E-test Agar dilution method Broth dilution method 3. Detection of resistance genes (vana, vanb, meca, teta,tetb., tetm etc..)

11 RESEARCH ACTIVITY D i c e ( O p t: 2. % ) (T o l 1. % - 1. % ) ( H >. % S >. % ) [. % -1. % ] s m a I I 65 6 sm a 1. Staphylococcosis Study of virulence factors, PFGE and analysis of protein A polymorphism of Staphylococcus aureus isolates (molecular epidemiology) C V -A C V -A C V -A C V -A C V -A 2. Enteric diseases Quali-quantitative study of the rabbit intestinal microbiota through Real-Time PCR 11

12 3. Animal welfare Study of environmental parameters in intensive rabbit farms and their effects on animal health and welfare Noxious gas (ammonia, carbon dioxide, methane, hydrogen sulphide) Bacteria and fungal spores 12

13 3. Animal welfare Study of environmental parameters in intensive rabbit farms and their effects on animal health and welfare Microclimatic parameters Temperature, relative humidity, air speed Total dust 13

14 Fresh air inlets Animal welfare Study of environmental parameters in intensive rabbit farms and their effects on animal health and welfare Exhaust air outlets Fresh air inlets 25 Exhaust air outlets (air extraction fans) Ammonia Carbon dioxide Time (hours) 15 Ammonia and carbon dioxide concentration trends over a 7-day period 5 Fresh air inlets Ammonia concentration patterns in different ventilation systems NH3 (mg/m 3 ); CO2 (g/m 3 )

15 3. Animal welfare Study of environmental parameters in intensive rabbit farms and their effects on animal health and welfare 25 2 middle inlet side external Air temperature ( C) Time (hours) Air temperature in different points inside a rabbit house compared to external temperature over a 7-days period 15

16 4. Clostridial enteric diseases Set up of biomolecular methods for the detection identification characterization (toxinotyping, ribotyping) of clostridia 16

17 Fabrizio Agnoletti Istituto Zooprofilattico Sperimentale delle Venezie Treviso, Italy Treviso historic center and the new IZSVe laboratory 17