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编号	期刊名称	期卷号	日期	标题	作者	DOI	页码	摘要
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Sero-epidemiological study of the presence of hantaviruses in domestic dogs and cats from Belgium	A. Dobly, C. Cochez, E. Goossens, H. De Bosschere, ... P. Heyman	https://doi.org/10.1016/j.rvsc.2011.02.003	Pages 221-224	Hantaviruses are worldwide rodent-borne pathogens infecting humans and other animals mainly through inhalation of aerosols contaminated with rodent excreta. Few data are available on hantavirus serology and geographical distribution in dogs and cats. We therefore screened sera from pet dogs (N = 410) and cats (N = 124) in two regions of Belgium, using IgG ELISA and IFA. We analysed the effect of the owner’s address as well as pet gender and age on hantavirus status. Hantavirus antibodies were found in both species with a significantly higher seroprevalence in cats than in dogs (16.9% vs. 4.9%, P = 0.001). More dogs were infected in highly forested southern Belgium (harbouring more rodents) than in northern Belgium (10.5% vs. 3.0%, P = 0.002). In the south, hantavirus sero-positive cats were found in more densely forested localities than sero-negatives ones were (P = 0.033). These results are consistent with the ecological variations of hantavirus risks in humans.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Implications and challenges of tuberculosis in wildlife ungulates in Portugal: A molecular epidemiology perspective	Mónica V. Cunha, Filipa Matos, Ana Canto, Teresa Albuquerque, ... Ana Botelho	https://doi.org/10.1016/j.rvsc.2011.03.009	Pages 225-235	Mycobacterium bovis and, more rarely, Mycobacterium caprae, may cause zoonotic bovine tuberculosis (bTB) in an extensive range of animal species. In Portugal, during 2009, a remarkable raise of bTB incidence was registered in cattle along with an increase of new cases in wildlife. In this work, we reassess and update the molecular epidemiology of bTB in wild ungulates by including 83 new M. bovis and M. caprae isolates from wild boar and red deer obtained during 2008–2009. Spoligotyping identified 27 patterns in wild ungulates, including 11 patterns exclusive from deer and five from wild boar. The genetic relatedness of wildlife and livestock isolates is confirmed. However, the relative prevalence of the predominant genotypes is different between the two groups. Contrasting with the disease in livestock, which is widespread in the territory, the isolation of bTB in wildlife is, apparently, geographically localized and genotypic similarities of strains are observed at the Iberian level.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Evaluation of Spoligotyping and MIRU-VNTR for Mycobacterium bovis in Xinjiang, China	Zhaogang Sun, Rui Cao, Miao Tian, Xuxia Zhang, ... Chuanyou Li	https://doi.org/10.1016/j.rvsc.2011.03.012	Pages 236-239	Mycobacterium bovis, the causative agent of bovine tuberculosis (bTB) in cattle, is also a pathogen for human and other mammals. In this study, 406 cows were screened for bTB by both single intradermal comparative cervical tuberculin (SICCT) test and IFN-γ assay. 135 M. bovis were isolated from 31 SICCT and IFN-γ double-positive cows in Xinjiang Uygur Autonomous Region of China. Spoligotyping and MIRU-VNTR were evaluated for genotyping, and 4 and 7 genotypes were identified, respectively. A new combination of nine MIRU-VNTR loci was most discriminative for M. bovis clones from Xinjiang. Interestingly, two new spoligotypes (SB1903 and SB1904) and special repeat numbers of three loci (ETR-D, QUB 1895 and QUB 3336) were discovered in this study. These results indicated a specific epidemic conservation in Xinjiang, China. M. bovis strains with the unique genotypes were isolated from the herds maintaining parent cows imported from the bTB-free countries, suggesting a possible transmission from the local breed of Xinjiang brown cattle.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	cDNA cloning and recombinant expression of a proliferation inducing ligand (APRIL) from Père David’s deer	Cui Min, Qi Wang, Yu-Qing Chen, Shuang-Quan Zhang, ... Yi-Jun Ren	https://doi.org/10.1016/j.rvsc.2011.01.022	Pages 240-242	In the present study, the cDNA of Père David’s deer APRIL (miAPRIL) was successfully amplified, and nucleotide sequence analysis showed that the open reading frame (ORF) of miAPRIL consists of 753 bases encoding 250-amino acids. Phylogenetic analysis exhibited high homology with the even-toed ungulates (artiodactyla). The extracellular soluble domain of the miAPRIL (misAPRIL) fragment was cloned into the expression vector pET43.1a and transformed into Escherichia coli BL21 (DE3) for recombinant expression. SDS–PAGE and western blotting analysis indicated that misAPRIL protein expression was successfully achieved. Indirect immunofluorescence staining demonstrated that misAPRIL has the ability to bind to the surface of Père David’s deer lymphocytes. Flow cytometry analysis revealed that misAPRIL was able to enhance lymphocytes survival in a dose-dependent manner.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Polymorphism in GHRH gene and its association with growth traits in Chinese native cattle	Bao Zhang, Gaofeng Zhao, Xianyong Lan, Chuzhao Lei, ... Hong Chen	https://doi.org/10.1016/j.rvsc.2011.01.023	Pages 243-246	Growth hormone-releasing hormone (GHRH) is secreted by the hypothalamus and stimulates growth hormone (GH) released from the pituitary. Mutations detected in GHRH gene showed associations with animal production traits. The purpose of this study was to investigate the association of the GHRH gene with growth traits in Chinese native cattle. PCR-SSCP and sequencing were used to detect mutations of the GHRH gene in this study. One novel mutation 4251nt (C > T) was found and the frequencies of C allele were 0.8778 and 0.8476 for Qinchuan and Nanyang cattle, respectively. Body weight with the CT genotype was significantly higher (P < 0.05 or P < 0.01) than those with CC genotype for different growth periods (6, 12, 18, and 24 months old) in Nanyang cattle. Our findings suggested that polymorphism in bovine GHRH might be one of the important genetic factors to influence body weight.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Rumen fermentation and histology in light lambs as affected by forage supply and lactation length	J. ??lvarez-Rodríguez, E. Monleón, A. Sanz, J.J. Badiola, M. Joy	https://doi.org/10.1016/j.rvsc.2011.03.010	Pages 247-253	This study determined whether the rumen fermentation and histology traits may reflect the feeding strategy in light lambs (22–24 kg). Thirty-two single Rasa Aragonesa lambs were assigned to one of four treatments in a 2 × 2 factorial design. The factors were the inclusion of forage in the diet (alfalfa grazing vs. concentrate-fed indoors) and lactation length (weaning at 13 kg vs. suckling until slaughter). A multivariate canonical analysis discriminated individuals among feeding strategies. The main function differentiated weaned concentrate-fed lambs from the rest according to dorsal sac papillae height, ventral sac muscular layer thickness and the proportion of rumen valerate. The second function differentiated suckling concentrate-fed lambs from the rest according to plasma urea levels. Lactation length played an important role on rumen histology and protein utilization, especially in concentrate-fed lambs. Alfalfa grazing light lambs had similar rumen morphometric measures and fermentation characteristics, regardless of milk access.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Toxocara vitulorum in suckling calves in the Netherlands	F.H.M. Borgsteede, M. Holzhauer, F.L. Herder, E.G. Veldhuis-Wolterbeek, C. Hegeman	https://doi.org/10.1016/j.rvsc.2010.11.008	Pages 254-256	A Dutch farmer with beef cattle and suckling calves noticed one of the calves suffering from diarrhoea and observed large white worms near the perineum. In the faeces of this calf, eggs of Toxocara vitulorum were found. The majority of the cows in the herd were of the Piemontese breed previously imported from southern France. Treatment with albendazole was successful. Two other calves had positive faecal egg counts. One month after treatment of these calves with doramectin, no eggs were found. To prevent further infections on the farm and dissemination of this uncommon infection to other farms, the farmer decided to have all his cattle slaughtered.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Feline demodicosis caused by an unnamed species	Rui Kano, Ayako Hyuga, Jun Matsumoto, Sadao Nogami, ... Hiroshi Kamata	https://doi.org/10.1016/j.rvsc.2011.03.015	Pages 257-258	A case of feline demodicosis is described in this report. A 13-year-old spayed female domestic short hair cat weighing 4.5 kg was being treated with cefovecin and alternately with prednisone or methylprednisolone. On further physical examination, the cat showed mild erythema and hair loss on the bridge of the nose, around the eyes, on the chin, on the side part of the breast and on the abdomen. A large number of Demodex mites were found in deep skin scrapings from the affected areas. The cat was then treated with ivermectin at 600 μg/kg administered SC daily. After 4 weeks of treatment, the cat was clinically normal with no mites detected in the skin scrapings from the face or breast areas. The mite responsible may represent a previously seen but as yet unnamed new species. This is third report that describes a case of feline demodicosis caused by a different, unnamed mite species that has different morphological characteristics to those of known Demodex mites and may represent a previously seen but as yet unnamed species.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Molecular cloning, expression and characterization of enolase from adult Haemonchus contortus	Kaikai Han, Lixin Xu, Ruofeng Yan, Xiaokai Song, Xiangrui Li	https://doi.org/10.1016/j.rvsc.2011.03.008	Pages 259-265	Enolase represents a multifunctional protein involved in basic energy metabolism. In the present research, the enolase gene of Haemonchus contortus (HcENO) was cloned and characterized. Specific primers for the rapid amplification of cDNA ends (RACE) were designed based on the expression sequence tag (EST, GenBank Accession No. BF422728) to amplify the 3′- and 5′-ends of HcENO. The full length of cDNA from this gene was obtained by overlapping the sequences of 3′- and 5′-extremities and amplification by reverse transcription PCR. The biochemical activities of the recombinant protein HcENO, which was expressed in prokaryotic cells and purified by affinity chromatography, were analyzed by assays of enzymatic activity, stability to pH. The results showed that the cloned full length cDNA comprised 1583 bp and encoded a peptide with 434 amino acid residues which showed sequence similarity to several known enolases. The biochemical assay showed that the protein encoded by the HcENO exhibited enzymatic activity, whilst the HcENO was stable between pH 6 and 8. The natural enolase of H. contortus detected by immunoblot assay was approximately 49 kDa in size, and the recombinant HcENO was recognized strongly by serum from experimentally infected goats.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Perosomus elumbis, cerebral aplasia, and spina bifida in an aborted Thoroughbred foal	I. Gerhauser, F. Geburek, P. Wohlsein	https://doi.org/10.1016/j.rvsc.2010.11.009	Pages 266-268	Perosomus elumbis represents a rare congenital anomaly characterized by aplasia of the lumbosacral spinal cord and vertebrae. This anomaly is often associated with arthrogryposis and malformations of the urogenital and intestinal tract. This report describes the first case of perosomus elumbis in an aborted Thoroughbred foal associated with cerebral aplasia with meningocele, cranioschisis, spina bifida, a fused urogenital and intestinal tracts lined by a cutaneous mucosa without uterine glands, atresia ani, and arthrogryposis of the hind legs. Immunohistochemistry detected no abnormalities in the GFAP, S-100, vimentin, NeuN, doublecortin, and neurofilament expression pattern in the remnants of the brainstem. Pathogenic bacteria or equine herpesviruses one and four were not found using a microbiological investigation or polymerase chain reaction, respectively. The observed malformations may be induced by a severe developmental disturbance during the formation of the neural tube with secondary distortion of the surrounding mesoderm. Although the aetiology of this dysorganogenesis remains undetermined, inherited chromosomal mutations, an intake of different xenobiotics, and/or a disturbed metabolism of the mare may have caused the defect.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Matrix metalloproteinase expression in sheep with listerial meningoencephalitis	Fatma ??lhan, Yavuz Ulusoy, Mehmet Hal??gür	https://doi.org/10.1016/j.rvsc.2011.01.019	Pages 269-272	Matrix metalloproteinases (MMPs) have been implicated in the pathogenesis of several central nervous system (CNS) diseases. In this study, we investigated the presence of Listeria monocytogenes antigens and detected the expression of MMP-9 and MMP-7 in the brains of 22 sheep with clinical signs and histopathological findings characteristic of listerial meningoencephalitis. Archived sections from the brainstem, cerebrum, and cerebellum were stained for immunohistochemistry. L. monocytogenes antigens were located mainly in the cytoplasm of neutrophils and some macrophages and/or extracellularly within microabscesses of the brainstem. MMP-9 was mainly immunolocalised in the endothelial cells, microglial cells, and neurons especially in inflammatory areas. MMP-7 immunoreactivity was detected in perivascular cuffs, microglial cells, and only a few neurons. Overall, immunohistochemistry in formalin-fixed, paraffin-embedded tissues is a useful tool for the diagnosis of encephalitic listeriosis caused by L. monocytogenes, and MMP-9 and MMP-7 may contribute to the pathogenesis of listerial meningoencephalitis.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Clinical and morphological investigations on the incidence of forms of rickets and their association with other pathological states in broiler chickens	I. Dinev	https://doi.org/10.1016/j.rvsc.2011.02.011	Pages 273-277	The purpose of investigations was to determine, by means of pathomorphological methods, the incidence of different rickets forms and their role for the occurrence of other illnesses in broiler chickens in Bulgaria.Clinical, blood biochemical, gross anatomy and histological investigations were carried out in broiler chickens with signs of rickets. The studies were performed in 12 broiler flocks in 4 farms located in different regions of the country. Based on macro- and microscopic lesions, alterations specific for hypocalcaemic rickets were observed in two farms, whereas signs of hypophosphataemic rickets – in the other two. The rickets diagnosis was confirmed by analysis of blood serum calcium, magnesium and inorganic phosphate concentrations.At the age of 30–35 days, various pathological states were observed in the same farms. The presented results suggested that existing problems in studied flocks were associated with an earlier occurrence of rickets.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	The efficacy and safety of levobupivacaine administered by lumbosacral epidural route in conscious sheep	Rafael DeRossi, Amadeu B. Silva-Neto, Cassio T.D. Pompermeyer, Fabrício O. Frazílio, ... Andrea C.L. de Barros	https://doi.org/10.1016/j.rvsc.2010.11.006	Pages 278-282	The aim of this study was to evaluate 3 doses of levobupivacaine (LB) epidurally administered in sheep. Six adult male 24–36 month-old sheep received levobupivacaine at 3 doses, LB05 (0.05 mg/kg), LB15 (0.15 mg/kg), and LB25 (0.25 mg/kg), and saline solution into the lumbosacral epidural space. Heart rate, arterial blood pressure (systolic, diastolic, and mean), respiratory rate, rectal, and skin temperature, local anesthesia, and ataxia were determined before treatment and at predetermined intervals. The duration of local anesthesia was 30 ± 5 min, 145 ± 27 min, and 290 ± 18 min for LB05, LB15, and LB25, respectively (P < 0.05). Ataxia determined for LB05, LB15, or LB25 was similar to the anesthetic times. There was an increase in heart rate and reduction in arterial pressure in LB25 (P < 0.05), whereas LB05 or LB15 did not affect these parameters. Lumbosacral epidural levobupivacaine is an appropriate choice for local anesthesia in sheep.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Cross-sectional study of hepatic CYP1A and CYP3A enzymes in hybrid striped bass, channel catfish and Nile tilapia following oxytetracycline treatment	N. Topic Popovic, T. Howell, J.G. Babish, P.R. Bowser	https://doi.org/10.1016/j.rvsc.2011.03.003	Pages 283-291	Terramycin for Fish?? (oxytetracycline, OTC) is one of three approved drugs for therapeutic treatment of fish in the United States. Nothing is known, however, of the effects of this therapeutic on drug metabolizing enzymes in fish post-treatment. The main purpose of the study was to examine whether the fish CYP1A and CYP3A enzymes would cross-react with antibodies to known mammalian cytochrome P-450 forms (CYP1A1 and CYP3A). Observational feeding studies of OTC effects were conducted in hybrid striped bass, channel catfish and Nile tilapia. Oxytetracycline was mixed into the feed to achieve a daily dose of 82.8 mg per kg body weight at a feeding rate of 1% body weight per day. Hepatic microsomes of each fish were prepared and Western blotting of CYP1A1 and CYP3A4 and enzyme assays of CYP1A2 and CYP3A4 were performed prior to OTC treatment and on post-treatment days 1, 6, 11 and 21. Both goat anti-rat CYP1A1 and rabbit anti-human CYP3A4 showed good cross-reactivity with all three species in this study. All three species exhibited distinct perturbations in one or more of the variables examined on day 1 post-treatment. Immediately following the 10-day medication period, relative liver weight (RLW) of hybrid striped bass was increased 44% and remained elevated through post-treatment day 21. Increased CYP3A4 enzyme activity and protein abundance were noted in channel catfish and Nile tilapia, respectively. This observational approach demonstrated species differences both in control activities and in the timing and extent of hepatic responses to OTC. The unique perturbations of hepatic CYP450 enzymes in different fish species to OTC treatment observed in this study may have relevance for the use of additional antibiotics or other therapeutics used in aquaculture.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Biochemical properties of bull spermatozoa separated in iodixanol density solution	Blanka Beer-Ljubi??, Jasna Aladrovi??, Terezija Silvija Marenjak, Ivanka Maji??-Bali??, ... Suzana Milinkovi??-Tur	https://doi.org/10.1016/j.rvsc.2011.01.011	Pages 292-294	Bull spermatozoa samples contain variable portion of motile and normal morphology spermatozoa along with spermatozoa incapable of fertilization due to their pathologic changes. As semen quality is influenced by biochemical and morphological characteristics of all spermatozoa, the aim of the study was to separate spermatozoa in discontinuous iodixanol density gradient solution and to determine their cholesterol, phospholipid, triacylglycerol and lipid peroxide concentrations and creatine kinase activity. The study was performed in winter and included seven Simmental bulls aged 1.5–3.5 years. Semen samples were collected by use of artificial vagina. Upon evaluation of semen quality (volume, concentration and progressive sperm motility), the samples were centrifuged in iodixanol density solution to obtain two sperm fractions. The two fractions included sperms with progressive motility greater than 90% and less than 20%, respectively. A statistically significantly higher lipid peroxide concentration was determined in sperm fraction with <20% progressive motility. Different sperm subpopulations can be obtained by separating bull spermatozoa in different iodixanol density gradient solutions, while monitoring their biochemical properties can help assess the sperm quality.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Ultrasonography of the abomasum in 30 Saanen goats	U. Braun, D. Jacquat	https://doi.org/10.1016/j.rvsc.2011.03.013	Pages 295-298	This report describes the findings of ultrasonographic examination of the abomasum in 30 goats. A 5.0 MHz linear transducer was used to scan the left and right paramedian regions, the linea alba and the right abdominal wall of standing goats. The abomasum appeared as a heterogeneous, moderately echogenic structure with echogenic stippling. The abomasum could be seen from the linea alba in all the goats. The abomasal folds were easily identified in 19 goats and appeared as prominent echogenic bands. The cranial visible limit of the abomasum was ??0.3 2.34 cm from the xyphoid, and the caudal limit was 10.3 3.08 cm caudal to the xyphoid. The visible length of the abomasum was 10.7 1.96 cm and the maximum width 30.2 7.43 cm. The height of the abomasum was 6.7 1.79 cm.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Effect of hypoxia/reoxygenation on the contractility of the isolated bovine digital vein	A. Risso, S. Comerma-Steffensen, J. Rojas, E. Ascanio, ... H. Zerpa	https://doi.org/10.1016/j.rvsc.2010.11.007	Pages 299-305	The bovine digital vasculature contractility has been implicated in the development of laminitis. To investigate the effect of hypoxia/reoxygenation on the contractility of isolated peripheral bovine digital veins (BDVs), vessel rings were studied under isometric conditions and submitted to 30 min of hypoxia (95%N2–5%CO2) and reoxygenation (95%O2–5%CO2) conditions, respectively. The BDVs contracted with a high K+ depolarizing solution, developed hypoxia-induced relaxation, followed by an increase in tension upon reoxygenation. In contrast, phenylephrine-contracted BDVs displayed a rapid, sustained and reversible hypoxia-induced contraction. Reoxygenation caused a rapid relaxation in phenylephrine-contracted BDVs. The presence of the endothelium did not modify the hypoxia/reoxygenation effects and hypoxia-induced contraction was still observed in a nominal Ca2+-free Krebs, however, the last effect was not maintained over time. The hypoxia-induced contraction in an isolated peripheral vein may contribute to the understanding of the physiology and pathophysiology of superficial venous smooth muscle contractility, particularly in the alteration of bovine digital haemodynamics under hypoxia/reoxygenation conditions.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Immunomodulation during and after castration under inhalation anaesthetic without genotoxic effects on equine lymphocytes	A. Strasser, H. Kühnel, K. Velde, A. Dadak	https://doi.org/10.1016/j.rvsc.2011.01.021	Pages 306-310	Genotoxic DNA damage due to inhalation anaesthesia has been demonstrated in human lymphocytes. In order to evaluate anaesthesia-associated changes in cell-mediated immunity on the basis of a potential DNA damage as a health risk in horses, single cell gel electrophoresis and lymphocyte proliferation assay were performed on equine lymphocytes which were obtained before, during and after regular castration under inhalation anaesthetic. No significant lymphocytic DNA damage due to isoflurane anaesthesia was observed, whereas lymphocyte proliferative reactivity and lymphocyte counts decreased significantly (p ?? 0.05) during and after anaesthesia. The present study thus indicates that the combined anaesthesia does not result in significant DNA damage, which hence cannot be held responsible for the observed changes in the immune response of equine lymphocytes. However, the recognized compromises of immune function ought to be considered especially in immunologically challenged animals.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Osteoclast progenitors from cats with and without tooth resorption respond differently to 1,25-dihydroxyvitamin D and interleukin-6	H.E. Booij-Vrieling, T.J. de Vries, T. Schoenmaker, M.A. Tryfonidou, ... V. Everts	https://doi.org/10.1016/j.rvsc.2011.03.014	Pages 311-316	Both vitamin D and inflammatory cytokines can stimulate osteoclast formation and activity. We studied the effect of 1,25-dihydroxycholecalciferol (1,25(OH)2D), and interleukin-6 (IL-6), on the formation and activity of feline osteoclasts, using peripheral blood mononuclear cells (PBMCs) from cats with and without tooth resorption (TR+ and TR??) as a source of osteoclast precursors. The formation of osteoclast-like cells (defined as multinucleated, tartrate-resistant acid phosphatase-positive cells) was assessed at 7 and 14 days. In the presence of M-CSF and RANKL, with and without IL-6, more osteoclasts were formed from TR?? PBMCs than from TR+ PBMCs on plastic. More osteoclasts were formed from TR+ PBMCs on bone slices in the presence of M-CSF/RANKL with 1,25(OH)2D. This opposite effect may be due to a higher expression of the vitamin D receptor (VDR) in TR+ osteoclasts and precursors on bone. Formation of resorption pits was analyzed and confirmed with scanning electron microscopy. In conclusion, we propose that TR+ PBMCs when cultured on bone are sensitive to 1,25(OH)2D, whereas the differentiation of TR?? PMBCs on bone seem more sensitive to IL-6, suggesting that osteoclast precursors from cats with and without tooth resorption respond differently to osteoclast stimulating factors.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Experimental evidence for competitive growth advantage of genotype VII over VI: Implications for foot-and-mouth disease virus serotype A genotype turnover in nature	J.K. Mohapatra, S. Subramaniam, N.K. Singh, A. Sanyal, B. Pattnaik	https://doi.org/10.1016/j.rvsc.2011.01.030	Pages 317-319	In India, systematic genotype replacement has been observed for serotype A foot-and-mouth disease virus. After a decade of co-circulation of genotypes VI and VII, genotype VII emerged as the single dominant genotype since 2001. To derive possible explanations for such epochal evolution dynamics, in vitro intergenotype growth competition experiments involving both co- and superinfection regimes were conducted. Coinfection of BHK-21 cells demonstrated abrupt loss in the genotype VI viral load with commensurate increase in the load of genotype VII as measured by the genotype differentiating ELISA, RT-PCR and real-time RT-PCR. The superinfection dynamics was shaped by temporal spacing of infection, where the invading genotype VII took more number of passages than coinfection to eventually overtake the resident genotype VI. It was speculated that such superior replicative fitness of genotype VII could have been a possible factor for the ultimate dominance of genotype VII in nature.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Antiviral activity of Alpinia katsumadai extracts against rotaviruses	Ha-Hyun Kim, Hyung-Jun Kwon, Young Bae Ryu, Jong Sun Chang, ... Woo Song Lee	https://doi.org/10.1016/j.rvsc.2010.11.012	Pages 320-323	In vitro anti-rotavirus activity of Alpinia katsumadai (AK) extracts were evaluated against bovine G8P7 and porcine G5P7 rotaviruses in two different assay strategies, a mixed treatment assay and a post treatment assay. In the mixed treatment assay, six AK extracts AK-1 (EtOH extract), AK-3 (H2O layer), AK-5 (40% methanol fraction), and AK-9–11 (H2O extract, polysaccharide fraction, supernatant fraction) exhibited inhibitory activities against G5P7 rotavirus with the EC50 values ranging from 0.7 ± 0.4 to 33.7 ± 6.5 μg/mL. Extracts AK-1, AK-3, and AK-5 inhibited rotavirus infection against G8P7 rotavirus, the with EC50 values of 8.4 ± 2.2 μg/mL, 6.5 ± 0.8 μg/mL and 8.4 ± 5.0 μg/mL, respectively. By hemagglutination inhibition (HI) assay, six AK extracts completely inhibited viral adsorption onto human RBCs in both strains of rotaviruses at less than 11 μg/mL. However, in the post treatment assay, there was no anti activity shown against both strains of rotaviruses. As a result, six AK extracts were attributed mainly to having a strong interaction with hemagglutinin protein on the outer surface of rotavirus, resulting to blockage of viral adsorption.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	First genetic characterization of equine adenovirus type 1 (EAdV-1) in Turkey	Veysel Soydal Ataseven, Tuba ??i??dem O??uzo??lu, Zeynep Ba??aran-Karap??nar, Seval Bilge-Da??alp	https://doi.org/10.1016/j.rvsc.2011.03.001	Pages 324-326	Equine adenovirus type 1 (EAdV-1) is a cause of repiratory tract infection in equids. In present study for the first time in Turkey, the prevalence of EAdV-1 in nasal swab samples obtained from horses showing respiratory symptoms was investigated by polymerase chain reaction (PCR), and molecular characterization of the hexon gene detected in the Turkish (TR) strain was performed. Overall, the prevalence of EAdV-1 was found low (1.4%) as indicated by a positive PCR reaction from the nasal swab extracts tested. Phylogenetic analysis based on the partial sequences of the hexon gene of a TR-EAdV-1 strain with those of previously isolated AdVs from different mammals and an EAdV-1 M1 strain showed that the EAdV-1 strains were placed into a unique cluster. Although the TR-EAdV-1 strain was closely related to CAV-1, CAV-2 and bat adenovirus reference strains, larger-scale studies are necessary to better understand the molecular epidemiology and population structure of EAdV-1 in Turkey.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Induction of feline immunodeficiency virus from a chronically infected feline T-lymphocyte cell line	Tadafumi S. Tochikura, Yuko Naito, Yasunori Kozutsumi, Tsutomu Hohdatsu	https://doi.org/10.1016/j.rvsc.2011.02.013	Pages 327-332	The infection of the feline T-lymphocyte cell line FeT-J with the feline immunodeficiency virus (FIV) Petaluma strain led to the establishment of nonvirus-producing cells. One clone (C15) obtained by limiting dilution was found to express FIV in response to chemical inducers of retroviruses. The chemical treatment of C15 cells led to not only FIV protein synthesis but also an augmentation of viral production. Examination of the C15 cell derivatives obtained by recloning revealed that 10–40% of treated cells constitutively expressed FIV antigens, whereas 100% with expressed FIV antigen in response to the inducer. Chemical induction resulted in more than a 100-fold increase in infectious viral production. The results suggest that a majority of FeT-J cells that are infected with FIV exist in a non-productive state. Establishing a cell line that can be non-productively infected by FIV may help determine the mechanisms of FIV latency.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Cowpox virus isolate virulent in humans shows attenuated phenotype in mice	Hartwig P. Huemer, Caroline Lassnig, Norbert Nowotny	https://doi.org/10.1016/j.rvsc.2011.03.011	Pages 333-337	We have cultured Cowpox virus (CPXV) from skin lesion material of a human patient from Austria. Phylogenetic comparison of the HA-gene revealed a rather homogeneous cluster with other local isolates from recent years, the A36R-gene was mostly related to elephant derived strains from Germany. Despite causing disease in human, the isolate AT/Carinthia/788/07 surprisingly even at high titers showed a highly reduced virulence in BALB/c mice upon intranasal inoculation as compared to vaccinia virus. This contrasts earlier reports on other CPXV isolates. Using shotgun DNA sequencing several insertions and deletions were found in genes presumably involved in host range, immune regulation as well as established virulence factors. These preliminary data could be an indication that CPXV strains with proven pathogenicity for humans may have reduced virulence in mice and vice versa. Additionally strains with a reduced virulence may have an advantage in persisting in less dense rodent populations.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Effects of hot-iron disbudding, using regional anaesthesia with and without carprofen, on cortisol and behaviour of calves	G. Stilwell, M.S. Lima, R.C. Carvalho, D.M. Broom	https://doi.org/10.1016/j.rvsc.2011.02.005	Pages 338-341	The objective of this study was to assess cortisol and behaviour changes in calves hot-iron disbudded after different analgesic protocols. We assessed the response of 27 calves at 1, 3, 6 and 24 h after disbudding with regional anaesthesia (DA), regional anaesthesia plus carprofen (DAC), disbudded only (D) or sham-disbudded (ND). At 1 h cortisol was higher in D compared with all other groups. At 3 h DA showed higher cortisol than ND but did not differ from baseline. At 15 min and 1 h pain-related behaviours were more frequent in the D group than in all other groups. Group D (3 h) and DA (3 and 6 h) showed more ear flicks and head rubs compared with DAC and ND. We concluded that head rubbing, head shaking and ear flicking are useful behaviours for evaluating pain after hot-iron disbudding. Disbudding causes severe pain in calves and only the association of regional anaesthesia with carprofen assures good welfare for 24 h.
857	Research in Veterinary Science	Volume 92, Issue 2	20120401	Lactic acid bacteria isolated from young calves – Characterization and potential as probiotics	Natalia C. Maldonado, Clara Silva de Ruiz, María Claudia Otero, Fernando Sesma, María Elena Nader-Macías	https://doi.org/10.1016/j.rvsc.2011.03.017	Pages 342-349	Lactic acid bacteria (LAB) are widely used as probiotics in humans and animals to restore the ecological balance of different mucosa. They help in the physiological functions of newborn calves that are susceptible to a variety of syndromes. The criteria for the selection of strains for the design of probiotic products are not available. Based in the host-specificity of the indigenous microbiota, 96 LAB isolates from faeces and oral cavity of calves were obtained. The surface properties were screened showing a small number of highly hydrophobic or autoagglutinating isolates. Also, a group produced H2O2 and were able to inhibit pathogens, and two strains were bacteriocin-producers. Some grew at very low pH and high bile concentrations.The strains sharing some of the specific properties evaluated were identified genetically, assayed their compatibility and exopolysaccharide production. The results allow going further in the establishment of criteria to select strains to be included in a multi-strain-probiotic-product to be further assayed in animals.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Editorial Board		https://doi.org/10.1016/S0034-5288(11)00476-0	Page i
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Endothelial hyperpolarizing factor increases acetylcholine-induced vasodilatation in pulmonary hypertensive broilers arterial rings	Diana I. Alvarez-Medina, Aureliano Hernandez, Camilo Orozco	https://doi.org/10.1016/j.rvsc.2011.02.004	Pages 1-6	Pulmonary arterial hypertension (PAH) develops as result of imbalances between endothelium derived vasoconstrictors and vasodilators. Pulmonary hypertensive broiler chickens (PHBs) are deficient in NO production and endothelin-1 (ET-1) excess. With respect to prostacyclin, it appears that it does not alter vascular pulmonary tone in broilers. However, the role of Endothelium-Derived Hyperpolarizing Factor (EDHF) in PAH in broilers has not been determined. The possible involvement of EDHF in acetylcholine (Ach) induced vasodilatation was studied in pulmonary arterial rings taken from PHB and non-pulmonary hypertensive broilers (NPHBs). Ach induced higher vasodilatation in PHB than in NPHB. This dilatation seems to be directly related to the degree of PAH. Ach derived vasodilatation was inhibited, in PBH but not in NPHB, by blocking EDHF action with K+ or Apamin plus Charybdotoxin. It is proposed EDHF as an important vasodilator in the pulmonary arteries of PHB, which may play a compensatory role in PAH pathophysiology.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Computed tomography measurements of thoracic structures in 26 clinically normal goats	Stefanie Ohlerth, Mareike Becker-Birck, Heinz Augsburger, Rahel Jud, ... Ueli Braun	https://doi.org/10.1016/j.rvsc.2010.10.019	Pages 7-12	The present study was performed to provide computed tomographic (CT) reference values for structures in the thorax of 26 clinically normal Saanen goats. Animals were anesthetized, positioned in sternal recumbency and transverse images with a reconstructed 1.5 mm slice thickness were obtained by use of a 40-slice CT scanner. Absolute and relative measurements of the trachea, heart, cranial vena cava, thoracic aorta, caudal vena cava, right and left principal bronchus, right and left caudal lobar bronchus and the concomitant branch of the right and left pulmonary artery and vein, large caudal mediastinal lymph node and lung density were performed with dedicated software. Minimal to moderate interstitial or bronchopneumonia was incidentally found on CT in 24 animals. In conclusion, CT images obtained in this study can be used as a reference for the evaluation of thoracic diseases in goats.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	California mastitis test scores as indicators of subclinical intra-mammary infections at the end of lactation in dairy cows	A.L. Bhutto, R.D. Murray, Z. Woldehiwet	https://doi.org/10.1016/j.rvsc.2010.10.006	Pages 13-17	Intramammary infections (IMI) during the dry period can be reduced through the use of dry cow therapy (DCT); in the future, its blanket use is likely to be questioned in the light of public concern regarding the routine use of antibiotics in food producing animals. One possible alternative is to limit DCT to cows with IMI just before drying off, which would require a quick, simple identification of sub-clinical IMI. In the present study we examined quarter milk samples obtained from 240 cows one week before and on the day of drying off, using the California mastitis test (CMT) and for IMI by bacteriological culture. The results indicated that high CMT scores at drying off may be good indicators of IMI: there was a significant association between the frequency of isolation of major pathogens and the CMT score in milk samples obtained one week before (Pearson’s χ2 = 27.04, df = 4, p < 0.001) and those at drying off (Pearson’s χ2 = 25.87, df = 4, p < 0.001).
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Determination of flagellar types by PCR-RFLP analysis of enteropathogenic Escherichia coli (EPEC) and Shiga toxin-producing E. coli (STEC) strains isolated from animals in S??o Paulo, Brazil	Claudia de Oliveira Ayala, Ana Carolina Ramos Moreno, Marina Baquerizo Martinez, Ylanna Kelner Burgos, ... Silvia Yumi Bando	https://doi.org/10.1016/j.rvsc.2010.10.025	Pages 18-23	This study evaluated the polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) analysis of fliC for typing flagella antigen (H) of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) strains isolated from different animals. The molecular typing of the H type was efficient in the determination of 93 (85%) strains. Two nonmotile (H-) E. coli strains showed a PCR-RFLP electrophoretic profile that did not match known H type patterns. The fliC nucleotide sequence of strains B2N and 4a revealed a nucleotide substitution at the restriction site and a nucleotide insertion that generated a stop codon, respectively. The results of this study showed that PCR-RFLP analysis of fliC is faster, less laborious and as efficient for the determination of H type E. coli isolated from animals, compared to serotyping and that it is useful in determining H type in nonmotile strains and strains expressing non-reactive H antigens. Moreover, the fliC sequence of strain B2N suggests that we could have found a new flagellin antigen type.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Tei index (myocardial performance index) and cardiac biomarkers in dogs with parvoviral enteritis	Meric Kocaturk, Silvia Martinez, Oya Eralp, Asta Tvarijonaviciute, ... Zeki Yilmaz	https://doi.org/10.1016/j.rvsc.2010.10.018	Pages 24-29	Tei index (myocardial performance) and cardiac biomarkers were evaluated in dogs with parvoviral enteritis (PVE). Tei index was calculated as isovolumic contraction time plus isovolumic relaxation time divided by ejection time. Myocardial and skeletal muscle damages were assessed by serum levels of cardiac troponin I (cTnI), creatine (phospho) kinase, lactate dehydrogenase and aspartate aminotransferase. Serum magnesium level was also determined. According to treatment response, dogs were divided into the survivor (n = 20) and non-survivor groups (n = 23). Seven healthy dogs served as controls. The mean value of the Tei index was higher in non-survivors, compared with survivors (p < 0.02) and healthy controls (p < 0.01). Serum level of cTnI in non-survivors was higher than that of survivors and controls (p < 0.05). Tei index showed the highest sensitivity and specificity to predict mortality. The findings of an elevated Tei index and an increase in serum cTnI are factors associated with a poor prognosis in cases of canine parvovirosis.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Fatal bovine anaplasmosis in a herd with new genotypes of Anaplasma marginale, Anaplasma ovis and concurrent haemoplasmosis	S. Hornok, A. Micsutka, I.G. Fernández de Mera, M.L. Meli, ... J. de la Fuente	https://doi.org/10.1016/j.rvsc.2010.10.011	Pages 30-35	Haematological and molecular analysis of blood samples was carried out during an outbreak of bovine anaplasmosis in Hungary. Acute disease was observed in five animals, two of which died. Anaplasma-carrier state was diagnosed in 69 (92%) of cattle. Further evaluation of 24 blood samples revealed concurrent infections with Mycoplasma wenyonii and ‘Candidatus M. haemobos’ in 22 and 21 animals, respectively. In addition, two cows were identified with rickettsaemia. Regarding molecular investigation of potential hard tick vectors, Haemaphysalis inermis and Dermacentor marginatus males collected from the animals were PCR-negative. However, in one pool (out of 18) of Ixodes ricinus males, and in six pools (out of 18) of D. reticulatus males the msp4 gene of Anaplasma marginale was detected. In the same I. ricinus pool Anaplasma ovis was also identified. All ticks were negative for haemoplasmas. Anaplasma sequences yielded 97–99% homology to sequences deposited in the Genbank. This is the first report of fatal bovine anaplasmosis associated with divergent A. marginale genotypes and concurrent ‘Candidatus M. haemobos’ infection, as well as of an A. ovis strain in ticks collected from cattle.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Seroprevalence of canine herpesvirus-1 in Turkish dog population	Kadir Ye??ilba??, Ebru Yal????n, Pelin Tuncer, Zeki Y??lmaz	https://doi.org/10.1016/j.rvsc.2010.10.016	Pages 36-39	Canine herpesvirus-1 (CHV-1) is the agent of reproductive and respiratory disorders in adult dogs, and the infection generally results in haemorrhagic disease conditions and neonatal death. In this study, virus neutralisation test that used complement (VNT) as well as in-house ELISA were utilised to investigate the CHV-1 seroprevalence in the Turkish dog population. Among the 560 serum samples, 39.3% of the samples tested by ELISA were CHV-1 positive while 29.4% of the samples tested by VNT were CHV-1 positive. Compared to the individual dogs (39.0%), there was a higher CHV-1 seroprevalence (62.1%) found in the colony dogs (62.1%) (p = 0.0002). However, there was an insignificant difference between male and female dogs. Although the highest antibody prevalence (56.7%) was found in Golden Retrievers, there were no significant variations detected among the dog breeds used in this study. Neutralizing antibody titres were very low (??1:16) in a high portion of the tested animals, confirming the rapid decrease of CHV-1 antibodies after the course of infection. The results of this study show that CHV-1 seroprevalence is moderately high in the Turkish dog population.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Effects of hyperimmune equine plasma on clinical and cellular responses in a low-dose endotoxaemia model in horses	G. Forbes, S. Church, C.J. Savage, S.R. Bailey	https://doi.org/10.1016/j.rvsc.2010.10.020	Pages 40-44	Endotoxaemia is a major cause of equine morbidity, and plasma from horses immunised against Escherichia coli is used in its treatment. The aim of this study was to determine the effects of hyperimmune plasma on the clinical and leukocyte responses, including production and activity of TNFα, in an in vivo endotoxin challenge model. Pre-treatment with hyperimmune equine plasma had no significant effect on peak total plasma TNFα concentration (occurring 90 min after the administration of 30 ng/kg LPS). However, the bioavailable (unbound) TNFα measured by bioassay was significantly reduced in plasma-treated horses (1044.44 ± 193.93 pg/ml at 90 min) compared to saline treated controls (1373.92 ± 107.63 pg/ml; P = 0.05). Therefore, although pre-treatment with hyperimmune equine plasma did not significantly modify the clinical signs of endotoxaemia in this model, there was some evidence of reduced TNF bioactivity, which may be due to factors in the plasma which bind and reduce the activity of this cytokine.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Molecular identification and characterization of the intervening sequences (IVSs) within 23S ribosomal RNA (rRNA) genes of Taylorella asinigenitalis isolated in France	Akihiro Tazumi, Sandrine Petry, Kyohei Hayashi, John E. Moore, ... Motoo Matsuda	https://doi.org/10.1016/j.rvsc.2010.10.004	Pages 45-52	In the helix 25 region, 32 French Taylorella asinigenitalis isolates carried at least one 23S rRNA gene not containing intervening sequences (IVSs). No IVSs in the region were identified in three isolates and the other remaining 29 isolates carried one or more IVSs (UCD-1TIVS1A, UCD-1TIVS1B and UK-1IVS1B) described already and two new kinds of IVS (TaIVS1C and TaIVS1D). In the helix 45 region, no T. asinigenitalis isolates not carrying any IVSs were identified. UK-1IVS2B was identified in the region from 26 isolates. Five new kinds of IVSs (TaIVS2D, E, F, G and H) occurred in the region in the 13 isolates. Distinctly different tandem repeat units (RS48 and RS32 and RS-A, -B and -C) were evident in both regions, respectively, from the French (n = 32) and American (n = 3) T. asinigenitalis isolates. Thus, several different kinds of tandem repeat units and their combinations in IVSs in both regions within the gene were shown in 32 French isolates.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Term placenta shows methylation independent down regulation of Cyp19 gene in animals with retained fetal membranes	Sandeep Ghai, Rachna Monga, T.K. Mohanty, M.S. Chauhan, Dheer Singh	https://doi.org/10.1016/j.rvsc.2010.10.008	Pages 53-59	Retention of fetal membranes (RFM) is the major post-partum disorder in dairy cattle. Cyp19 gene encodes the aromatase enzyme responsible for catalyzing the rate limiting step in estrogen biosynthesis, an important hormone for placental maturation and expulsion. The present study was aimed for comparative analysis of Cyp19 gene expression and its epigenetic regulation in placental cotyledons of animals with and without RFM. Significantly lower expression of Cyp19 gene was found in placental samples of RFM affected animals in comparison to normal animals. Methylation analysis of 5 CpG dinucleotides of placenta specific Cyp19 gene promoter I.1 and proximal promoter, PII showed hypo-methylation of both PI.1 and PII in term placenta of normal and diseased animals. In conclusion, a mechanism other than promoter methylation is responsible for decreased aromatase expression in placental cotyledons of animals suffering from RFM.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Transcriptional profiling of antimicrobial peptides avian β-defensins in the chicken ovary during sexual maturation and in response to Salmonella enteritidis infection	G. Michailidis, M. Avdi, A. Argiriou	https://doi.org/10.1016/j.rvsc.2010.10.010	Pages 60-65	Avian β-defensins (AvβDs) are antimicrobial peptides that play significant roles in the innate immune system in chickens. The aim of this study was to identify the types of AvβDs expressed in the chicken ovary, to investigate the effects of sexual maturation in the ovarian mRNA abundance and to determine the changes in their expression levels as a result to Salmonella enteritidis (SE) infection. RNA was extracted from the ovary of healthy prepubertal, sexually mature and aged birds, as well as from sexually mature and aged SE infected birds. Real-time PCR analysis revealed that 11 AvβDs genes were expressed in the chicken ovary. A significant up regulation of AvβD1, 3, 4, 5, 7 and 11 was observed in the ovary of sexually mature and aged birds. Furthermore, a significant up-regulation of AvβD4, 5, 7, 11 and 12 was observed in the ovary of SE infected sexually mature birds. These results suggest that the mRNA expression of at least six AvβDs increase with age in the ovary of laying hens, and that at least five AvβDs show an induction in their expression in response to SE infection, indicating an AvβD-mediated immune response mechanism in the chicken ovary.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Osteogenic proliferation and differentiation of canine bone marrow and adipose tissue derived mesenchymal stromal cells and the influence of hypoxia	Dai-Jung Chung, Kei Hayashi, Chrisoula A. Toupadakis, Alice Wong, Clare E. Yellowley	https://doi.org/10.1016/j.rvsc.2010.10.012	Pages 66-75	The aim of this study was to compare the osteogenic and proliferative potential of canine mesenchymal stromal cells (cMSCs) derived from bone marrow (BM-cMSCs) and adipose tissue (AT-cMSCs). Proliferation potential was determined under varying oxygen tensions (1%, 5%, and 21% O2). Effects of reduced oxygen levels on the osteogenic differentiation of AT-cMSCs were also investigated. AT-cMSCs proliferated at a significantly faster rate than BM-cMSCs, although both cell types showed robust osteogenic differentiation. Culture in 5% and 1% O2 impaired proliferation in cMSC from both sources and osteogenic differentiation in AT-cMSCs. Our data suggests that AT-cMSCs might be more suitable for use in a clinical situation, where large cell numbers are required for bone repair, due to their rapid proliferation combined with robust osteogenic potential. Our data also suggests that the inhibitory effects of hypoxia on both cell proliferation and differentiation should be considered when using MSCs in a potentially hypoxic environment such as a fracture site.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Genetic variation of major histocompatibility complex BLB2 gene exon 2 in Hebei domestic chicken	Xiu-Li Guo, Hui-qin Zheng, Xiang-Long Li, Ying Li, ... Lan-Hui Li	https://doi.org/10.1016/j.rvsc.2010.10.021	Pages 76-79	Genetic variation of MHC BLB2 gene exon 2 in Hebei domestic chicken was investigated, after PCR and sequencing of a 374 bp fragment (containing entire exon 2 (270 bp) of BLB2 gene) in 76 individuals. The results showed that along this fragment, there were 69 variable sites, of which 18 were novel variations, and 82 estimated haplotypes with the diversity of 0.960. In Hebei domestic chicken, the nucleotide diversity (π), the average number of nucleotide differences (k), the average number of nucleotide diversity of synonymous substitution (π(s)) and non-synonymous substitution (π(a)) in BLB2 gene exon 2 were 0.098, 24.688, 0.075, and 0.106, respectively; nine non-synonymous substitutions was exclusively found in the peptide-binding sites (PBS) region of BLB2 gene exon 2, inferring that these unique substitutions might be helpful to resist some special bacteria and pathogens. The higher genetic diversity of MHC BLB2 gene exon 2 in Hebei domestic chicken might be consistent with its more robust disease resistance.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Calreticulin expression in neoplastic versus normal dog mammary glands: A cDNA subtraction-based study	Takumi Okawa, Yuzo Kurio, Masahiro Morimoto, Toshiharu Hayashi, ... Takuya Mizuno	https://doi.org/10.1016/j.rvsc.2010.10.028	Pages 80-91	This is the first report describing the expression of canine calreticulin (cCRT) in canine mammary gland tumour (MGT). Using cDNA subtraction method, it is found that mRNAs of CRT, cathepsin A, ovostatin, and lactotransferrin were differentially expressed in mammary adenocarcinoma as compared to hyperplasia, both of which were obtained from the dog. Furthermore, the mRNA expression levels of CRT and cathepsin A were significantly higher in canine MGT samples than in nontumour samples. In contrast, immunohistochemical studies have indicated that the expression of cCRT protein found to be detected in most of mammary gland tissues and was not correlated to the types of canine MGTs. Furthermore, cCRT was molecularly cloned, and the amino acid sequence of cCRT was found to be very similar to those of other species. Further studies are required to elucidate additional roles of cCRT in canine MGT.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Isolation, propagation, identification and comparative pathogenicity of five Egyptian field strains of Eimeria tenella from broiler chickens in five different provinces in Egypt	Somaia Saif Abu-Akkada, Ashraf Mahmoud Awad	https://doi.org/10.1016/j.rvsc.2010.10.023	Pages 92-95	A battery trial was conducted to evaluate the comparative pathogenicity of five field strains of Eimeria tenella from Behera, Khafr El-Sheikh, Alexandria, Gharbia and Matrouh provinces in Egypt. Two-week-old broiler chickens were infected with 25 × 103 sporulated oocysts of each strain of E. tenella. The comparative pathogenicity of the strains was assessed by calculating body weight gain, feed conversion ratio, lesion scores, dropping nature scores, cecal scrapings, mortality percentage and oocysts count. Hematological parameters including hemoglobin (Hb) content, packed cell volume (PCV%) and total erythrocytic count, were also evaluated. There were different degrees of pathogenicity between the strains.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Detection of specific antibodies anti-Neospora caninum in the fallow deer (Dama dama)	Justyna Bień, Bo??ena Moskwa, Marek Bogdaszewski, W??adyslaw Cabaj	https://doi.org/10.1016/j.rvsc.2010.10.014	Pages 96-98	Sera from 335 farmed fallow deer (Dama dama) at the breeding station in Kosewo Górne in the Mazurian Lake District, North-East Poland, were investigated for the presence of antibodies against Neospora caninum. The distribution of age groups was as follow: >4 years – 154 animals; 2 years – 76 animals; 1 year – 105 animals.Ten sera with the optical density exceeding 0.159 absorbance units (i.e., cut-off value) in ELISA test were also analyzed by Western blot. Western blot analysis revealed seroreactivity against immunodominant N. caninum antigens of 37, 25, and 16 kDa; however, in some sera additional bands were also visible.This is the first screening studies for antibodies against N. caninum in farmed fallow deer in Poland, in the region where neosporosis was confirmed in cattle and in farmed and free-ranging European red deer (Cervus elaphus).
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Characterization of Bunostomum trigonocephalum and Bunostomum phlebotomum from sheep and cattle by internal transcribed spacers of nuclear ribosomal DNA	C.R. Wang, J.F. Gao, X.Q. Zhu, Q. Zhao	https://doi.org/10.1016/j.rvsc.2010.10.024	Pages 99-102	In the present study, samples representing Bunostomum trigonocephalum and Bunostomum phlebotomum from sheep and cattle in Heilongjiang Province, China, were characterized and grouped genetically by the first (ITS-1) and second (ITS-2) internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA). The rDNA region including the ITS-1, 5.8S, ITS-2, and flanking 18S and 28S rDNA sequences was amplified by polymerase chain reaction (PCR), then sequenced and compared with that of other members of the hookworms available in GenBank??, and phylogenetic relationships between them were reconstructed using the Maximum-Parsimony method. The ITS-1, 5.8S, and ITS-2 sequences of the sheep hookworm were 381, 153, and 231 bp in length, respectively, and the corresponding sequences of the cattle hookworm were 392, 153, and 240 bp in length. The identity of ITS sequences of B. trigonocephalum and B. phlebotomum from sheep and cattle was 87.4%. A PCR-linked restriction fragment length polymorphism (PCR-RFLP) assay using restriction endonuclease Nde I was established for the unequivocal differentiation of the two hookworm species. Phylogenetic analyses based on the ITS sequences revealed that B. trigonocephalum and B. phlebotomum were closely related, but they represent two different species.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	2-Aminoethoxydiphenyl borate (2-APB) reduces respiratory burst, MMP-9 release and CD11b expression, and increases l-selectin shedding in bovine neutrophils	I. Conejeros, E. Jara, M.D. Carretta, P. Alarcón, ... R.A. Burgos	https://doi.org/10.1016/j.rvsc.2010.10.005	Pages 103-110	This study describes the effect of 2-aminoethoxydiphenyl borate (2-APB), a putative store-operated calcium (Ca2+) entry (SOCE) inhibitor, on reactive oxygen species (ROS) production, matrix metalloproteinase 9 (MMP-9) release, CD11b and l-selectin (CD62L) expression, size changes and apoptosis in bovine neutrophils stimulated with platelet-activating factor (PAF). It was observed that doses ??1 μM 2-APB significantly reduced ROS production, whereas 50 and 100 μM 2-APB reduced MMP-9 release induced by PAF. Moreover, concentrations ??10 μM 2-APB reduced CD11b expression and increased l-selectin shedding. PAF induced size changes in neutrophils, and this effect was inhibited by 2-APB. From this work it is possible to conclude that 2-APB at concentrations that inhibit SOCE responses was able to inhibit ROS and MMP-9 release and CD11b expression, and increase l-selectin shedding, suggesting that the Ca2+ channel involved in SOCE is a potential target for the development of new anti-inflammatory drugs in cattle.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Pharmacokinetics in pulmonary epithelial lining fluid and plasma of ampicillin and pivampicillin administered to horses	Lotte Winther, Keith Edward Baptiste, Christian Friis	https://doi.org/10.1016/j.rvsc.2010.11.001	Pages 111-115	Ampicillin concentrations in pulmonary epithelial lining fluid (PELF) and plasma was studied after single intravenous ampicillin administration (15 mg/kg) or single intragastric administration of its prodrug, pivampicillin (19.9 mg/kg) to horses and discussed in relation to minimum inhibitory concentrations (MIC) of common equine respiratory pathogens.After intravenous administration, elimination of ampicillin was fast and not detectable in plasma after 12 h in three out of six horses. Pivampicillin was absorbed well in non-fasted horses with an oral bioavailability of 36%. The degree of penetration of ampicillin into PELF, as described by the AUCPELF/AUCplasma ratio from 0 to 12 h was 0.40 after intravenous administration and 1.00 after pivampicillin administration.In horses, ampicillin administered either intravenously or orally, in the form of pivampicillin, can provide clinically relevant drug concentrations in PELF for at least 12 h, when treating susceptible equine respiratory pathogens (e.g. streptococci). Treatment of other bacterial pathogens requires susceptibility testing and possibly more frequent dosing, depending of minimum inhibitory concentrations (MIC) values.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Effects induced by exercise on lymphocyte β-adrenergic receptors and plasma catecholamine levels in performance horses	B. Cuniberti, P. Badino, R. Odore, C. Girardi, G. Re	https://doi.org/10.1016/j.rvsc.2010.11.002	Pages 116-120	The effect of dynamic exercise on complete blood cell count, lymphocyte β-adrenergic receptor and plasma catecholamine (adrenaline and noradrenaline) levels in horses performing different disciplines were investigated during rest and after exercise.Blood samples were collected from jumping horses (n = 6), Arabian Endurance horses (n = 6) and Standardbred trotters (n = 6) before and immediately after competition. Dynamic exercise caused a significant increase in red blood cell count (Standardbred trotters: P = 0.0012), haemoglobin concentration (jumping horses: P = 0.001; Standardbred trotters: P = 0.01), haematocrit percentage (Standardbred trotters: P = 0.005), neutrophil percentage (jumping horses: P = 0.0003), lymphocyte percentage (jumping horses: P = 0.0003), monocyte percentage (Standardbred trotters: P = 0.0008), lymphocyte β-AR numbers (jumping horses: P = 0.01; Arabian Endurance horses: P = 0.016; Standardbred trotters: P = 0.05), plasma adrenaline concentration (Standardbred trotters: P = 0.0001) and plasma noradrenaline levels (Standardbred trotters: P = 0.003). It is concluded that acute increases in plasma catecholamine concentrations depended on the exercise performed and may induce up-regulation of β-AR in equine lymphocytes. However, the exact mechanism of β-AR up-regulation still remains unclear.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Factors affecting rectal temperature measurement using commonly available digital thermometers	Jonathan M. Naylor, Renee M. Streeter, Paul Torgerson	https://doi.org/10.1016/j.rvsc.2010.10.027	Pages 121-123	Rectal temperature measurement is an essential part of physical examination of cattle and some physiological experiments. Modern digital thermometers are often used to measure rectal temperatures by students; this study describes their reliability and appropriate use. Students measured rectal temperature on 53 occasions using their personal digital thermometer and techniques gained from previous instruction, rectal temperature was also measured by an experienced person using a Cornell mercury thermometer completely inserted in the rectum. Cornell mercury thermometers values were 38.95 ± 0.05 °C (mean ± 1 SE, n = 53). Student rectal temperature measurements using their initial technique were nearly 0.5 °C lower, 38.46 ± 0.07 °C. After receiving instruction to insert the digital thermometer to the window, student obtained values were 38.77 ± 0.06 °C; these are significantly higher than with the student’s initial technique and closer to those obtained with a Cornell thermometer. In a series of 53 water bath tests, student owned thermometers recorded similar mean values to those of a traceable (reference) digital thermometer, Cornell mercury thermometer readings were 0.2 °C higher. 10 individual digital thermometers were repeatedly tested against a traceable thermometer in a water bath, one was inaccurate. In a separate experiment a trained clinician tested the effect of angle of insertion of a digital thermometer on temperature readings and the affect was <0.1 °C. We conclude that accurate temperature measurements using digital thermometers are only likely if the thermometer is inserted to the beginning of the window and the thermometer’s accuracy is checked periodically.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Effect of melatonin on DNA damage of bovine cumulus cells during in vitro maturation (IVM) and on in vitro embryo development	L. Takada, A. Martins Junior, G.Z. Mingoti, J.C.C. Balieiro, ... L.A. Coelho	https://doi.org/10.1016/j.rvsc.2010.11.004	Pages 124-127	The effect of melatonin during in vitro maturation (IVM) on DNA damage of cumulus cells (CCs) from bovine cumulus–oocyte complexes (COCs) and embryo development was evaluated. COCs from abattoir ovaries were cultured in maturation medium (MM) with 0.5 μg/ml FSH and 5.0 μg/ml LH (FSH–LH); 10??9 M melatonin (MEL) or FSH–LH + MEL (FSH–LH–MEL). After 24 h of in vitro maturation, the CCs surrounding the oocyte were subjected to DNA analysis by Comet assay. After in vitro fertilization and in vitro embryo culture, the embryo development rates were evaluated on day 2 post insemination (cleavage) and days 7–8 (blastocyst). The percentage of CCs with no DNA damage was significantly superior in MEL group (37.6 ± 2.4) than in FSH–LH–MEL (28.0 ± 2.4) and FSH–LH (17.8 ± 2.41) groups. Cleavage and blastocysts rates were similar among groups. Melatonin during IVM protects the CCs from DNA damage but this effect did not influence embryo development in vitro.
857	Research in Veterinary Science	Volume 92, Issue 1	20120201	Haematological values of post-laying Arrau turtle (Podocnemis expansa) in the Orinoco River, Venezuela	M. Rossini, P.A. Blanco, E. Marín, S. Comerma-Steffensen, H. Zerpa	https://doi.org/10.1016/j.rvsc.2010.10.026	Pages 128-131	The Arrau turtle (Podocnemis expansa) is an endangered species, as a result of long-lasting, unsustainable exploitation. To obtain reference haematological values from the wild Podocnemis expansa during post-laying, 20 turtles were captured in the Orinoco River. Blood was obtained from the dorsal cervical sinus in lithium heparin tubes. Red blood cells (RBC), white blood cells (WBC), thrombocytes (TC), packed cell volume (PCV), plasmatic protein (PP), haemoglobin (Hgb), mean corpuscular volume (MCV) and differential leukocyte count were determined. Haematological values were: RBC 0.9 × 109/L, WBC 5.7 × 109/L, TC 5.4 × 109/L, PCV 35.6%, PP 4.2 g/dL, Hgb 11.8 g/dL, MCV 411 fL. The differential leukocyte count comprised: 71% heterophils, 23% lymphocytes, 3% eosinophils, 1.6% basophils, and 1% monocytes. The reports of reference haematology values for the wild P. expansa are limited; therefore, the results presented herein contrast with those values obtained in captivity. This study represents a contribution to the referential haematological values of the wild P. expansa.

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