SELECTED ABSTRACTS OF THE 5TH ANNUAL MEETING OF THE ISRAEL ASSOCIATION OF VETERINARY MICROBIOLOGY AND IMMUNOLOGY, JANUARY 2009

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SELECTED ABSTRACTS
SELECTED ABSTRACTS OF THE 5TH ANNUAL MEETING OF THE ISRAEL ASSOCIATION OF VETERINARY MICROBIOLOGY AND IMMUNOLOGY, JANUARY 2009
Chairman: Elad, D.
DO ANTI- PORPhIROMONAS LEVII ANTIBODIES IN PREPARATORY HEIFERS AFFECT THE OCCURRENCE OF BOVINE NECROTIC VULVUVAGINITIS POST-PARTURITION?
Zamir, L. 1, Blum, S. 1, Leitner, G. 1, Krifuchs, O. 1, Goshen, T. 2, Brenner, J. 1 and Elad, D. 1 1 Div. of Bacteriology, Kimron Veterinary Institute, Bet Dagan, 2Hahaklait, Caesarea. Bovine necrotic vulvuvaginitis (BNVV) is an important syndrome in some dairy herds in Israel, which can potentially lead to sever economic losses (3). Primiparous cows are at the highest risk. Clinical signs appear three days after calving, starting with erythematic lesions that progress to hemorrhages leading to inflammation and necrosis of the vaginal mucosa. Severe cases of BNVV can result in peritonitis and death. Thus far there is no successful treatment of BNVV(1). BNVV was first diagnosed in 2001 in several dairy herds in the north of Israel and subsequent additional herds in the country became affected. All the infected herds had a history of introducing a large number of cows during consolidation with other farms during unification with small herds. The etiological agent of the syndrome is Porphyromonas levii, a Gram-negative anaerobe, commensal of the digestive tract that colonizes mucosal lesions of the post-parturient vagina. The microorganism is unable to cause BNVV without the help of additional predisposing factors. The principal factor is assumed to be stress associated with animal transfer and calving (4). However, BNVV has become endemic on two farms, with new cases appearing for several years after the consolidation of the farm. This indicates that factors other than the aforementioned stresses are necessary for predisposition to the syndrome. The aim of this study was to characterize the humoral response to P. levii during the periparturient period in order to assess whether a correlation exists between the presence of anti-P. levii antibodies before parturition and the appearance of BNVV afterwards.The study was conducted on three herds: A-endemic (54 cows), B-outbreak (51 cows) and C-unaffected (17 cows). Serum samples were taken on the weeks around days 210, 240 and 270 of pregnancy, and the first and fourth weeks post- parturition. ELISA tests were conducted to determine the levels and type of anti- P. levi antibodies in each sample.
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Antibodies were found in 59/105 (56.2%) heifers preparturition, indicating previous exposure. No correlation was found between the presence of antibodies preparturition and development of BNVV. However, only four (28.6%) of 14 cases with the high preparturion titers, developed BNVV after parturition. In contrast, among cows with lower or no titer, 41.9% (36/86) developed BNVV. The humoral pattern of clean herds was similar to infected ones. Also, heifers that developed BNVV had a higher mean antibody titer after calving than healthy animals. In conclusion, the presence of antibodies before parturition does not explain the difference between infected and uninfected herds and between afflicted and healthy heifers in the infected herds. However, the relatively low number of BNVV-positive heifers among the animals with high preparturient antibody titer, indicates that an prevention of the syndrome by vaccination would be justified. References: 1. Yeruham, I. Tiomkin, D. Friedgut, O. Van-Ham, M. Perl, S. and Elad, D.: Necrotic vulvovaginitis in dairy cattle in Israel. Vet. Rec. 160:164-166, 2007. 2. Elad, D. Friedgut, O. Alpert, N. Stram, Y. Lahav, D. and Tiomkin, D.: Bovine necrotic vulvovaginitis associated with Porphyromonas levii. Emerg Infect Dis. 10:505–507, 2004. 3. Blum, S. Mazuz, M. Brenner, J. Friedgut, O. Koren, O. Goshen, T. and Elad, D.: Effects of bovine necrotic vulvovaginitis on productivity in a dairy herd in Israel. Vet. J. 176 : 245–247, 2007. 4. Blum, S. Mazuz, M. Brenner, J. Friedgut, O. Stram, Y. Koren, O., Goshen, T. and Elad, D.: Sample-based assessment of the microbial etiology of bovine necrotic vulvovaginitis. Theriogenology. 68:290–293,2007. 5. Blum, S. Brenner, J. Friedgut, O. Stram, Y. Koren, O. Dagoni, I. Munbaz.A and Elad, D.: Porphyromonas levii in bovine necrotic vulvovaginitis negative dairy herds. Vet Record (accepted).
Volume 64 (1) 2009
website: www.isrvma.org
ISRAEL JOURNAL OF VETERINARY MEDICINE
THE FIRST EVIDENCE OF BVDV-2 IN AN ISRAELI HERD.
Stram, Y., Rubinstein-Guni, M., Kuznetzova L., Friedgut O and Rotenberg, D. Div. of Virology. Kimron Veterinary Institute, Bet Dagan Bovine viral diarrhea viruses belong to the pestivirus genus of the Flaviviridae. Analysis of genome sequences of BVD viruses distinguishes between two genotypes, BVDV-1 and BVDV-2. Antigenic differences occur between the genotypes, but there is considerable crossreactivity, so that pestiviruses are not classified into serotypes. The incursion of both genotypes of BVDV into a herd is usually inapparent and the main damage is the result of fetal infection. However there have been reports of severe clinical disease caused by virulent strains. The 5' UTR region of Israeli BVDV isolates, collected from 1996 to 2008 from various geographic locations, was sequenced and analyzed in order to characterize and map the isolates. BVDV-1 and -2 were both identified; BVDV-1 was the predominant genotype and could further be subtyped. Type 1b was the most common, but 1a and 1b were also found. An outbreak of severe acute BVDV infection occurred in an Israeli dairy herd in the summer of 2008. The outbreak presented 2 distinct clinical syndromes, characterized by hemorrhagic enteritis in adult cattle and severe pneumonia in young calves. Nine BVDV isolates from the outbreak were analyzed. All isolates from both clinical syndromes were identified as BVDV- 2 and were nearly identical phylogenetically.
USE OF IN VIVO INDUCED ANTIGEN TECHNOLOGY (IVIAT) FOR IDENTIFICATION AND CHARACTERIZATION OF MyCOPLASMA GALLISEPTICUM ANTIGENS PREFERENTIALLY EXPRESSED DURING INFECTION
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Ron M.1, Vaanunu E.1, Gorelik A.1, Lysnyansky I.2, Levisohn S.2 and Yogev D.1 Microbiology Institute, The Hebrew University, Jerusalem 91120 and 2Division of Avian and Aquatic Diseases, Five ivi-genes encode previously identified putative virulence factors of M. gallisepticum: phase variable VlhA lipoproteins (VlhA 1.07 and 4.01), the cytadhesin molecule GapA and two homologues of M. pneumoniae cytoskeletal proteins P65 and HMW3 (PlpA and Hlp3, respectively), which exhibit fibronectin binding capabilities. Identification of these known pathogenic determinants of M. gallisepticum by IVIAT validates this experimental approach. Interestingly, five additional IVIs proteins not previously implicated in infectivity were identified, including ABC predicted permease (MGA 0654), a hypothetical protein (MGA 0241), two 50S ribosomal proteins L23 and L2 (MGA 0711 and 0712), and a chaperonin GroEL protein. We believe that identification and characterization of ivi genes, which most likely encode virulence factors essential for survival, bacterial cell division and multiplication in situ, may provide new insights into M. gallisepticum pathogenesis as well as the development of efficacious vaccines, novel targets for antimicrobial therapy and innovative diagnostic tools.
Kimron Veterinary Institute, Bet Dagan, Israel Mycoplasma gallisepticum is known to cause chronic respiratory disease in chickens, infectious sinusitis in turkeys and is a major component of the respiratory complex in domestic poultry. Several virulence factors of M. gallisepticum have been characterized and shown to be involved in the pathogenicity of this organism. The focus of this study was to discover novel virulenceassociated determinants of M. gallisepticum induced in the host on infection, using in vivo induced antigen technology (IVIAT). In order to prepare antibodies detecting only in vivo induced (ivi) genes, serum samples from ten chickens experimentally infected with M. gallisepticum R (8p) were pooled and absorbed with the pathogen grown in vitro. An inducible expression library of M. gallisepticum R (25p) screened with anti-ivi serum yielded 10 positive clones. Identification of the ivi clones was performed by comparison of the nucleotide sequences of the mycoplasmal inserts with the available M. gallisepticum R genome sequence data. Analysis of the putative ivi clones revealed striking results.
Volume 64 (1) 2009
website: www.isrvma.org
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SELECTED ABSTRACTS
WHAT DO LUMPY SKIN DISEASE, PSEUDO LUMPY SKIN DISEASE AND ULCERATIVE HERPES MAMMILLITIS HAVE IN COMMON?
Brenner J. Kimron Veterinary Institute, POBox 12, 50250 Bet Dagan, Israel, yakovb@moag.gov.il BHV-2 belongs to the Alphaherpesviridae sub-family and produces mild to moderate disease in ruminants. It is probably transmitted mechanically while the other viruses belonging to this subfamily are transmitted by the respiratory route. Herpes viruses generally cause local diseases, with barely detectable levels of viremia. Bovine ulcerative herpes mammillitis (BHUM) and pseudo lumpy skin disease (PLSD), two clinically distinct diseases that have not been reported to appear on the same infected cow at the same time. BHUM is associated with a painful localized ulcerative udder, and although it seems to be present in Israel, it has not been reported officially. PLSD also known as Allerton virus infection is characterized as a generalized BHV-2 infection of cattle.. To the authors’ knowledge, the present outbreak is the fourth recorded case of PLSD outside Africa. In May 2008, a close examination of a primiparous dairy cow, presented lumps around and on the perianal region. The internal part of the tail revealed additional skin lesions. These were presented as scattered circular multifocal lesions, approximately 2 cm in diameter. Other lesions had lost hair and resembled ringworm lesions, were scattered among the lumps of the caudal region and on the dorsal region, between the neck and the hips, and were interspersed with a few wellcircumscribed hairless lesions. PLSD, as the generalized cutaneous form of BHV-2 infection does not seem to be a very serious disease in itself, but it raises concern at differential diagnosis between PLSD and LSDV infection, ringworm (Thricophyton verrucosum), and Dermatophilus congolensis infection. Lumpy skin disease (LSD) presents as an acute, subacute or latent infectious disease of cattle caused by a single strain of capripox virus known as Neethling virus. LSD is characterized by the rapid eruption of multiple circumscribed skin nodules, generalized lymphadenitis and fever. Although the field experience and circumstantial evidence suggest that transmission of LSDV occurs primarily by biting insects the mode of field transmission is not fully understood.
Table 1- Characteristics of PLSD, LSD and BUHM infections Clinical characterization Causal agent Anatomical appearance of the lesions Aspect of the lesion In between etiology Sharing intermediate clinical feature Differential diagnosis: PLSD BHV-2 Allerton virus Generalized skin lesions, caudal upper region mainly Lumps (pseudo) necrosis sloughs off? BHV-2 + Capripox Never appears with BUHM LSD LSD Capripox Neethling virus Generalized skin lesions, all over the body Lumps necrosis slough off BHV-2 ± Capripox + Lumps mixed with pseudo- lumps PLSD BUHM BHV-2 Allerton virus Local skin teat & udder only Granular ulcerative BHV-2 + Capripox Never appears with PLSD Unique
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website: www.isrvma.org
Volume 64 (1) 2009
ISRAEL JOURNAL OF VETERINARY MEDICINE
MICROBIOLOGY IN THE POST-GENOMIC ERA
Karnieli A., Karnieli Ltd., Molecular Diagnostic Division. Genomics has revolutionized every aspect of microbiology. Now, 13 years after the first bacterial genome was sequenced, it is important to pause and consider what has changed in microbiology research as a consequence of genomics. Over the past decade, genomic technologies have revolutionized microbiology and will probably continue to do so during the next decade. The information that is being added to sequence databases is increasing exponentially and every day we are in a better position to describe microorganisms by their genome, bacterial species by their pan-genome, and even complex microbial environments by their metagenomes. Advances in genomics are also beginning to drive the discovery of novel diagnostics, drug targets and vaccines. In this talk, I will review various aspects of the impact of genomics on microbiology in general and veterinary science in particular, including in the evolving field of bacterial typing, and the genomic technologies that enable the comparative analysis of multiple genomes. I will also address the implications of the genomic era for the future of veterinary microbiology and how this could assist and advance the veterinary practitioner in the day- to- day clinic.
CHARACTERIZATION OF QUINOLONE RESISTANCE-DETERMINING REGIONS IN MyCOPLASMA bOvIS CLINICAL ISOLATES WITH DIFFERENT SUSCEPTIBILITY TO ENROFLOXACIN
Lysnyansky I., Mikula I., Gerchman I. and Levisohn S. Division of Avian and Aquatic Diseases, Kimron Veterinary Institute, Bet Dagan 50250, Israel Emergence of resistance to fluoroquinolones is mainly due to chromosomal mutations in genes encoding the subunits of the drug’s target enzymes, DNA gyrase (GyrA and GyrB) and topoisomerase IV (ParC and ParE), which are essential for DNA replication. The quinolone resistance-determining regions (QRDRs) of these genes were characterized in 42 Mycoplasma bovis strains isolated between 2005 and 2008 from different clinical conditions and exhibiting various levels of susceptibility to fluoroquinolones. All enrofloxacinresistant isolates harbored amino acid substitutions in the QRDRs of each of three proteins (GyrA, ParC and ParE). However, the main difference between susceptible and resistant M. bovis strains was the change of nt G to A at position 265 of the parC amplicon, resulting in the substitution of 84-Asp/Asn. The parC-PCR-Restriction Fragment Length Polymorphism (RFLP) assay using PsiI restriction enzyme clearly differentiates between enrofloxacin resistant and susceptible M. bovis field strains. This feature may serve as a molecular marker for fluoroquinolone resistance. In addition, molecular typing of enrofloxacin resistant and susceptible M. bovis strains by insertion sequencelike elements-RFLP analysis, suggest that the currently prevalent enrofloxacin-resistant strain evolved by selection under field conditions from one of the susceptible strains.
Volume 64 (1) 2009
website: www.isrvma.org
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SELECTED ABSTRACTS
EMERGENCE OF DOG RABIES IN NORTHERN ISRAEL
David D.1, Dveres N.1, Davidson I.2 and Yakobson BA.1, 1 Rabies Laboratory and 2Division of Poultry Diseases, Kimron Veterinary Institute, POB 12, Bet Dagan, 50250, Israel Between the years 1979 to 2000, foxes constituted the main reservoirs of rabies in the Northern regions of Israel. Following the implementation of fox targeted oral vaccination programme (ORV) in 1998, rabies was eradicated from this area. Subsequently in the years 2004 to 2007, the biological and molecular characterization of the rabies isolates showed that stray dogs emerged as the main animal reservoir in Northern Israel while lower numbers of cases were reported in domestic animals. The virus isolates from foxes and dogs differed, in their molecular characterization, suggesting two distinct separate lineages. The transition from fox mediated rabies to dog-mediated rabies is a great concern to public health because of the close contact between dogs and the human population.
DETECTION OF bARTONELLA SPP. IN WILD RODENTS IN ISRAEL USING HRM REAL-TIME PCR.
Morick D.1, Baneth G.1, Avidor B.2, Mumcuoglu KY.3, Mintz D.1, Eyal O.1, Shpigel N.1 and Harrus S.1 1 Koret School of Veterinary Medicine, The Hebrew University of Jerusalem; 2Laboratory for Viruses and Molecular Biology, Tel-Aviv Sourasky Medical Center; 3Department of Parasitology, Hebrew University-Hadassah Medical School. The prevalence of Bartonella spp. in wild rodents was studied in four zoogeographical areas in Israel. One hundred and twelve rodents belonging to five species: Mus musculus, Rattus rattus, Microtus socialis, Acomys cahirinus and Apodemus sylvaticus were included in the survey. In addition, 152 external parasites belonging to three species: Pulex irritans, Xenopsyla cheopis and Haemaphysalis sp., and arthropod species belonging to one of the three following suborders: Anoplura, Gamasina and Ixodidae were collected from the rodents. DNA was extracted from spleen aspirates from each of the rodents and from pools of external parasites and examined for the presence of Bartonella spp. DNA. The diagnosis and molecular identification of Bartonella spp. was done by high resolution melt (HRM) real time PCR. The method, originally used for single nucleotide polymorphism (SNP) genotyping, was designed for the differentiation of 12 Bartonella spp. according to the nucleotide variation in two gene fragments, rpoB and gltA, and the 16S-23S intergenic spacer (ITS) locus simultaneously. Bartonella DNA was detected in spleen samples of 19 out of 79 (24%) black rats (R. rattus) and in 1 of 4 (25%) Cairo spiny mice (A. cahirinus). Bartonella DNA was detected in 15 of 34 (44%) flea-pools. All positive flea pools contained only rat-fleas (X. cheopis) and were collected from black rats (R. rattus). The Bartonella sp. detected in most samples was closely related to both B. tribocorum and the zoonotic B. elizabethae.
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website: www.isrvma.org
Volume 64 (1) 2009
ISRAEL JOURNAL OF VETERINARY MEDICINE
Cover image – Slipper lobster | Scyllarides latus
Slipper lobsters are a family of decapod crustaceans found in all warm oceans and seas. Despite their name, they are not true lobsters but are more closely related to spiny lobsters and furry lobsters, and they lack prominent claws. Slipper lobsters are easily identified by their enlarged antennae, which project forward from the head as wide plates. In the eastern Mediterranean their size rarely exceeds 40 cm.and they live on rocky substrates in depths up to 100 m. They are active at night and during the day they hide frequently inside caves and crevices. They feed on molluscs, mainly limpets and other sessile marine snails. The species has become rare in most parts of the Mediterranean Sea due to severe over-fishing for human food. In some places along Israel’s shores slipper lobsters can be spotted during the night, especially in the vicinity of artificial substrates like ship wrecks on the sea bed.
Volume 64 (1) 2009
website: www.isrvma.org
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