Abstract Book of EAVLD2012 - eavld congress 2012

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S3 - P - 25 VIRAL DIAGNOSIS USING TRANSMISSION ELECTRON MICROSCOPY SCHMALLENBERG VIRUS David J. Everest and W. A. Cooley 1 Central Animal Health and Veterinary Laboratories Agency-Weybridge, New Haw, Addlestone, Surrey KT15 3NB, UK Most viruses are very small and visualisation by Transmission Electron Microscopy (TEM) has provided a major contribution in the discovery and detection of viral infections. The AHVLA’s Bioimaging unit provides a rapid viral diagnostic service, analysing fresh unfixed scab, lesion, wart and faecal materials by TEM. Species range from domestic farm animals, to wildlife, marine mammals and Zoo animals, covering red squirrels to Elephants. Historically, this form of microscopy has been a key application in the diagnosis and surveillance of several wildlife viral infections. These have included Squirrel pox and adenovirus, both contributing to the decline of the UK Red Squirrel, calicivirus from Rabbits and Hares, the causative agent of rabbit haemorrhagic disease and pox viruses in various marine mammal species.
S3 - P - 26 DETECTION OF EQUINE FOAMY VIRUS INFECTIONS IN HORSES Magdalena Materniak, Jacek Kuzmak National Veterinary Research Institute, Department of Biochemistry, Pulawy, Poland Equine foamy virus, diagnostics, PCR, ELISA Introduction Foamy viruses (FVs) are the least known complex retroviruses. FVs infections are highly prevalent among several animal species including non-human primates (chimpanzees, gorillas, etc.), cats and cattle. Another foamy virus was isolated from horses (Equine foamy virus - EFV) (1). Similarly to other FVs, EFV shows characteristic ultrastructure and susceptibility to infect large number of cell lines. Furthermore, its molecular analysis exhibits genome organization typical for other known foamy viruses, but its sequence shows the highest similarity to bovine foamy virus. Although the first isolation of EFV was reported over ten years ago, still nothing is known about the prevalence of EFV infections in horses. Therefore we aimed to develop diagnostic tools and apply them to investigate the EFV infection in horses from Poland. References 1. Tobaly-Tapiero J., Bittoun P., Neves M., Guillemin M.-C., Lecellier Ch.- H., Puvion-Dutilleul F., Gicquel B., Zientara S., Giron M.-L., The H. de, Saϊb A. (2000). Isolation and characterization of an eqiune foamy virus. J. Virol., 74, 4064-4073. 2. Romen F., Backes P., Materniak M., Sting R., Vahlenkamp T. W., Riebe R., Pawlita M., Kuzmak J., Löchelt M. (2007). Serological detection systems for identification of cows shedding bovine foamy virus via milk. Virology, 364, 123-131. Materials & methods Blood samples were collected from 175 horses (4 -18 years old), including common breeds, Hucul ponies and randomly selected samples from slaughtered horses. Semi-nested PCR was developed and applied to detect EFV DNA in DNA extracted from peripheral blood leukocytes (PBLs). Selected primers flanked 275 bp fragment within pol gene, the most conservative EFV region. GST ELISA with BFV specific Gag-GST fusion protein (2) was used for detection of foamy virus specific antibodies in horse serum samples. To confirm ELISA results immunoblotting with antigen prepared from Cf2Th/BFV 100 cells was applied. Additionally, attempts were made to isolate EFV from peripheral blood leukocytes of PCR positive animals by co-cultivation with BHK-21 and RK-13 cells. Results 12.5% of samples were positive in semi-nested PCR. Specificity of amplified fragments was confirmed by sequencing and parwise identity of particular sequences ranged from 95.3% to 98.3%, when compared to EFV reference sequence (GenBank Acc. No. NC_002201). In contrast, almost 19% of the samples showed seroreactivity to BFV Gag protein. Most of PCR positive samples were detected in Hucul ponies, while only several in samples randomly collected from slaughtered animals. All attempts to isolate EFV from PBLs of PCR positive horses failed. Table 1: Summary of results. Origin PCR (+) ELISA (+) samples samples Hucul ponies 15 14 0 Saddle-horses 3 6 0 Slaughtered horses TOTAL 4 22 13 33 Co-cultivation (+) samples Discussion & conclusions This study presents the first report on prevalence of EFV infection in horses. The discordance between PCR and serological results can be due to application of BFV specific tests instead of EFV specific serological assays. Therefore further studies are essential to develop EFV specific serological tests to confirm obtained results. Nevertheless, our study clearly shows that EFV infections are present among horses in Poland. Acknowledgements The authors would like to thank Dr. Ali Saïb and Dr. Joelle Tobaly-Tapiero for providing EFV positive DNA and serum controls. 0 0
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2 nd CONGRESSOFTHEEUROPEAN ASSOCIAT
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Welcome Dear colleagues, Welcome to
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worked with many diagnostic compani
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Products and services for scientist
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16:4518.15 20:0023:00 [S1.]Oralpres
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Day4 Wednesday,4July2012 4 th sessi
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Oral presentations “General Sessi
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antigens or attenuated vaccines can
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conditions, e.g. Staphylococcus aur
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S1 - O - 1 ORAL FLUIDS - SAMPLE MAT
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S1 - O - 03 INTRODUCTION RATE OF A
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S1 - O -05 VALIDATION OF A COMPETIT
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S1 - O - 07 SUBTYPING OF SWINE INFL
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S1 - O - 09 VIRAL DIAGNOSIS USING T
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S1 - O - 11 USING ORAL FLUID FOR TH
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S1 - O - 13 THE FIRST YEAR OF OBLIG
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S1 - O - 15 SEROLOGICAL ANALYSIS AN
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S1 - O - 17 RING TEST EVALUATION FO
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Oral presentations “Diagnostics a
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S2 - O - 01 DEVELOPMENT OF A RAPID
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S2 - O - 03 EVALUATION OF RAPID HRS
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Oral presentations “Emerging, re-
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advances, the fundamental tools of
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S3 - O - 02 25 YEARS OF PASSIVE SUR
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S3 - O - 04 EQUINE NOCARDIOFORM PLA
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S3 - O - 06 DETECTION OF SCHMALLENB
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S3 - O - 08 SCHMALLENBERGVIRUS: SER
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S3 - O - 10 DIAGNOSTIC ASPECTS OF S
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Oral presentations “New technique
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acid and organic solvent. Then a dr
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S4 - O - 02 BIOLOG GENERATION III,
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S4 - O - 04 MATRIX-ASSISTED LASER D
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S4 - O - 06 15 MINUTE ELISA USING A
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S1 - P - 01 LAWSONIA INTRACELLULARI
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S1 - P - 03 DETECTION OF PRRSV ANTI
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S1 - P - 05 DEVELOPMENT AND EVALUAT
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S1 - P - 07 ANALYTICAL EVALUATION O
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S1 - P - 09 PTS FOR BVDV ANTIGEN DE
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S1 - P - 11 ANTIMICROBIAL SUSCEPTIB
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S1 - P - 13 IDENTIFICATION OF GLOBI
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S1 - P - 15 ACCURACY OF PARATUBERCU
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S1 - P - 17 VIROLOGICAL SURVEILLANC
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Abstract Book of EAVLD2012 - eavld congress 2012

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