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Equine rhinitis B virus 1 (ERBV1), genus Erbovirus, family Picornaviridae, is a pathogen of horses which causes clinical and subclinical infection of the upper respiratory tract in horses. The virus is widespread in European horse populations and the current standard method for the detection of antibody against ERBV1 is by virus neutralisation (VN). VN tests, however, are labour-intensive and time-consuming, require tissue culture facilities, and generally do not provide same-day results. In this study, a protocol for the high-level expression and purification of recombinant virion protein 1 (rVP1) was established using metal-chelate affinity chromatography under denaturing condition. When used as a coating antigen in a prototype enzyme-linked immunosorbent assay (ELISA), denatured rVP1 was recognised by ERBV1 antibody present in horse serum. This finding suggests that denatured rVP1 is a promising candidate for the development of an ELISA to be used in the routine laboratory diagnosis of ERBV1 infection in horses.
Kriegshauser, G. Cullinane, A. Kuechler, E. Skern, T.
Max F. Perutz Laboratories, Medical University of Vienna, Vienna, Austria.
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In 2004 and then in 2006 several outbreaks of infectious bursal disease (IBD) were reported in broiler and broiler breeder flocks in Slovenia. In this report ten recently emerged IBD viruses (IBDV) were characterised by sequence analysis of the VP2 hypervariable region and compared to previous Slovene IBDV strains from 1995/1996 and to some representative serotype 1 IBDV strains of different pathotypes. On the basis of nucleotide and amino acid identities, phylogenetic analyses and the presence of very virulent IBDV (vvIBDV) conserved amino acid substitutions, all Slovene isolates from recent outbreaks were identified as vvIBDV. Although some unique nucleotide exchanges and amino acid substitutions have been observed, the results of this study indicated that recent vvIBDV isolates are closely related with those from outbreaks in the 1990s. However, acute IBD has not been reported in commercial flocks in Slovenia for some years. This could lead to the conclusion that poor biosecurity and relaxed vaccination could be responsible for the re-emergence of vvIBDV.
Rojs, OZ Krapez, U. Slavec, B. Mankoc, S. Jurisic-Cizerl, R. Barlic-Maganja, D.
Veterinary Faculty, University of Ljubljana, Gerbiceva 60, 1000 Ljubljana, Slovenia.
The present study was designed to investigate the cryotolerance of in vitro fertilised (IVF) mouse embryos at various preimplantation developmental stages. IVF mouse embryos were vitrified by the open-pulled straw (OPS) method. After warming, embryos were morphologically evaluated and assessed by their development to blastocysts, hatched blastocysts or term. The results showed that a high proportion (93.3-100.0%) of vitrified embryos at all developmental stages were morphologically normal after recovery. The developmental rate of vitrified 1-cell embryos to blastocyst (40.0%) or hatched blastocyst (32.7%) or term (9.3%) was significantly lower than that from other stages (P<0.05). Vitrified embryos from 2-cell to early blastocyst stage showed similar blastocyst (71.8-89.5%) and hatched blastocyst rates (61.1-69.6%) and could develop to term without a significant loss of survival compared with those of fresh embryos (P>0.05). Vitrified 2-cell embryos showed the highest survival rate in vivo (50.6%, 88/174), compared with that from other stages (9.3-30.5%, P<0.05). The data demonstrate that the OPS method is suitable for the cryopreservation of IVF mouse embryos from 2-cell stage to early blastocyst stage without a significant loss of survival. Embryos at the 2-cell stage had the best tolerance for cryopreservation in the present study.
Yan ChangLiang Yang QiEn Zhou GuangBin Hou YunPeng Zhao XueMing Fan ZhiQiang Liu ManQing Liu Lin Zhu ShiEn
College of Animal Science and Technology, State Key Laboratories for Agrobiotechnology, China Agricultural University, Beijing, China.
The first weeks of lactation in dairy cows are characterized by elevated bone resorption. The connection between lactation and bone metabolism is still much discussed. In this work, changes in the concentration of plasma parathyroid hormone-related peptide (PTHrP) and markers of bone metabolism were studied in Holstein cows and heifers in the dry period and early lactation to determine the role of PTHrP in the relationship between the rate of bone remodelling and the onset of lactation in dairy cows. Blood samples were taken 14 days before calving (D-14, n=23) and then on day 10 (D+10, n=21) and day 30 after calving (D+30, n=23). Using enzyme immunoassay (EIA), the concentrations of PTHrP, parathyroid hormone (PTH), carboxyterminal cross-linked telopeptide of type I collagen (CTX) and oestradiol and the activity of bone specific alkaline phosphatase (BSALP) were determined. The results showed a statistically significant increase in plasma PTHrP (p<0.005) and CTX (p<0.0001) in cows on D+10 as compared to D-14 and CTX on D+30 as compared to D-14 (p<0.0001). Significant negative correlations were found between the concentrations of PTHrP and oestradiol (r=-0.29, p<0.05) and those of CTX and oestradiol (r=-0.54, p<0.0001). In nonpregnant heifers (n=6), the concentration of CTX and the activity of BSALP were significantly higher (p<0.0001) than in dry cows. The observed increments of PTHrP and bone resorption after parturition reveal adaptations of bone metabolism to lactation in dairy cows.
Filipovic, N. Stojevic, Z. Zdelar-Tuk, M. Kusec, V.
Department of Physiology and Radiobiology, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova 55, 10 000 Zagreb, Croatia.
Klebsiella (K.) oxytoca infection induced the abortion of a female equine fetus in the 10th month of pregnancy. Bacteria were cultured from the liver, lung and stomach content. They were labelled with an anti-Mycobacterium bovis antibody in the thymus, liver and lungs and were stained with Giemsa and Brown-Brenn staining in the thymus and lung. The diffusely consolidated lungs contained numerous grey-whitish foci 2-4 mm in diameter, which corresponded to severe pyogranulomatous pneumonia characterised by numerous intraalveolar neutrophils and macrophages and multinucleated Langhans’ giant cells. K. oxytoca was located in the cytoplasm of these cells, and extracellularly in the lumen of alveoli, bronchioles and bronchi, in the capsule of thymus and in the sinusoids of the liver. The results indicate that K. oxytoca can cause sporadic equine abortion.
Szeredi, L. Janosi, S. Tenk, M.
Central Agricultural Office, Veterinary Diagnostic Directorate, H-1149 Budapest, Tabornok u. 2, Hungary.
A postweaning pig died in spite of antibiotic therapy showing wasting in a small herd. Postweaning multisystemic wasting syndrome (PMWS) was diagnosed on the basis of gross pathological and histological lesions and the presence of moderate amounts of porcine circovirus 2 (PCV2) antigen in tissue samples. Mycotic gastritis caused by Zygomycetes spp. was found on round areas with a diameter of 1 to 3 cm in the glandular mucosa of the stomach. Moderate amount of PCV2 viral antigen was detected almost evently in the stomach and mostly in the macrophages. In addition, acute uraemia, revealed by an ammonia-like stink of the gastric mucosa and the presence of acute erosions on the glandular mucosa of the stomach, was observed as a consequence of PCV2-induced interstitial nephritis. Only PCV2 infection could be identified as a cause of secondary mycotic gastritis. The results further support the immunosuppressive ability of PCV2 infection in PMWS-affected pigs.
Szeredi, L. Szentirmai, C.
Central Agricultural Office, Veterinary Diagnostic Directorate, H-1149 Budapest, Tabornok u. 2, Hungary.
The aims of the present study were to determine: (a) the effectiveness of an attenuated live Cryptobia salmositica vaccine; (b) the effects of food deprivation on the immune response and its duration in rainbow trout (Oncorhynchus mykiss) immunized with a live C. salmositica vaccine or with a killed Aeromonas salmonicida vaccine. The fish were divided into three groups (I, II and III; 14 fish per group), those in Groups I and II were under food deprivation (0.40% of body weight), while Group III fish were fed to satiety. The study showed that the attenuated strain of C. salmositica did not cause anaemia and disease, and the fish were protected from clinical disease when they were challenged with virulent parasites. Parasitaemia in all fish vaccinated and challenged with virulent C. salmositica fluctuated and was relatively low; however, fish in Group III had higher parasitaemia than those in Groups I and II between weeks 8 and 14. The numbers of activated neutrophils increased [nitroblue tetrazolium (NBT) assay] after immunization with both Cryptobia and Aeromonas vaccines and they remained high throughout the experiment. Antibody production (ELISA values) increased after vaccination and were slightly higher in Group III. ELISA titres against A. salmonicida increased after vaccination and decreased after 5 weeks. The titres increased again after the vaccinated fish were given booster, and they were higher than those in the first vaccinated fish.
Nourollahi-Fard, SR Woo, P. TK
School of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran.
The goal of this study was to give insight into the lesser known action of neutrophils in the course of bovine respiratory disease (BRD). Neutrophils are principally involved in resistance to infection but under certain conditions these cells participate in lung injury. TNF alpha and IL-8, the primary cytokines which play essential roles in neutrophil activation during the inflammatory process, may lead not only to host defence reaction but also to destructive responses. Neutrophil constituents, such as elastase, myeloperoxidase (MPO), alkaline phosphatase (ALKP) and superoxide among others, contribute to tissue destruction. Also, 5-oxo-eicosatetraenoic acid (5-oxo-ETE), the potent activator of neutrophil chemotaxis and degranulation, can cause lung injury. This study revealed that neutrophils under the influence of TNF alpha exacerbated the release of elastase, MPO, ALKP and superoxide. IL-8, in turn, triggered those neutrophils which were isolated from heifers suffering from BRD, and released elastase, ALKP and 5-oxo-ETE, but not MPO or superoxide (O2-). The most pronounced degranulative response was observed in the acute form of BRD and a lesser response was seen in the chronic form. The secretory action of neutrophils varied not only depending on the form and phase of the disease but it also diminished successively during recovery.
Wessely-Szponder, J.
Department of Pathophysiology, Chair of Preclinical Veterinary Sciences, Faculty of Veterinary Medicine, Agricultural Academy, Akademicka 12, 20-033 Lublin, Poland.
The objective of this study was to estimate the effect of the thyroglobulin (TG) locus on beef quality traits in some beef cattle breeds and to investigate the effect of the DGAT1 locus on milk production traits in the Hungarian Holstein Friesian population. TG and DGAT1 genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. At the TG locus TT bulls showed the highest fat percentage values in the longissimus dorsi muscle (m. longissimus dorsi); the difference between CC and TT genotypes was significant. DGAT1 GC/GC cows had the highest milk, fat and protein yield values. Due to the relatively small number of GC/GC cows the difference proved to be significant only between AA/AA and AA/GC genotypes.
Anton, I. Kovacs, K. Fesus, L. Varhegyi, J. Lehel, L. Hajda, Z. Polgar, JP Szabo, F. Zsolnai, A.
Research Institute for Animal Breeding and Nutrition, H-2053 Herceghalom, Gesztenyes u. 1, Hungary.
Sensitisation to mites is frequent in atopic dogs. The main mite genus involved in canine atopic dermatitis is Dermatophagoides. The importance of storage mite allergens in dogs has been controversial. The aim of this study was to evaluate the sensitisation rates against storage mites (Lepidoglyphus destructor and Tyrophagus putrescentiae) and house dust mites (Dermatophagoides farinae and D. pteronyssinus) in atopic dogs from Galicia, a highly humid and temperate region of Spain, using a Fc epsilon RI alpha -based immunoglobulin E (IgE) in vitro test. The study was performed on 95 dogs suffering from atopic dermatitis and presenting detectable specific serum IgE levels: 91.6% of the dogs tested positive for storage mites, whereas sensitisation to house dust mites was detected in 87.4%. These results indicate the importance of storage mites in this specific geographic area.
Goicoa, A. Espino, L. Rodriguez, I. Puigdemont, A. Brazis, P. Rejas, J.
Department of Veterinary Clinical Sciences, Rof Codina Veterinary Teaching Hospital, Faculty of Veterinary Medicine, University of Santiago de Compostela, 22741 Lugo, Spain.