Polymerase chain reaction tests for the identification of Ross River, Kunjin and Murray Valley encephalitis virus infections in horses

OBJECTIVE: To develop and validate specific, sensitive and rapid diagnostic tests using RT-PCR for the detection of Ross River virus (RRV), Kunjin virus (KV) and Murray Valley encephalitis virus (MVEV) infections in horses. METHODS: Primer sets based on nucleotide sequence encoding the envelope glycoprotein E2 of RRV and on the nonstructural protein 5 (NS5) of KV and MVEV were designed and used in single round PCRs to test for the respective viruses in infected cell cultures and, in the case of RRV, in samples of horse blood and synovial fluid. RESULTS: The primer pairs designed for each of the three viruses amplified a product of expected size from prototype viruses that were grown in cell culture. The identity of each of the products was confirmed by nucleotide sequencing indicating that in the context used the RT-PCRs were specific. RRV was detected in serums from 8 horses for which there were clinical signs consistent with RRV infection such that an acute-phase serum sample was taken and submitted for RRV serology testing. The RRV RT-PCR was analytically sensitive in that it was estimated to detect as little as 50 TCID50 of RRV per mL of serum and was specific in that the primer pairs did not amplify other products from the 8 serum samples. The RRV primers also detected virus in three independent mosquito pools known to contain RRV by virus isolation in cell culture. Samples from horses suspected to be infected with KV and MVEV were not available. CONCLUSION: Despite much anecdotal and serological evidence for infection of horses with RRV actual infection and associated clinical disease are infrequently confirmed. The availability of a specific and analytically sensitive RT-PCR for the detection of RRV provides additional opportunities to confirm the presence of this virus in clinical samples. The RT-PCR primers for the diagnosis of KV and MVEV infections were shown to be specific for cell culture grown viruses but the further validation of these tests requires the availability of appropriate clinical samples from infected horses.     

Characterization of monoclonal antibodies against avian reovirus S1133 protein sigmaA synthesized in Escherichia coli

Monoclonal antibodies (MAbs) were prepared against avian reovirus S1133 protein sigmaA (esigmaA) synthesized in Escherichia coli. MAbs were characterized and used to develop a diagnostic test. Ten MAbs were selected for competitive binding assay following coupling with horseradish peroxidase. The results indicated that these MAbs delineated two epitopes I and II of esigmaA. An immuno-dot binding assay was used to detect the effect of denaturation on antibody recognition of the epitopes. All MAbs bound to esigmaA in its native form. After denaturation by boiling in SDS and 2-mercaptoethanol, the binding of MAbs recognizing epitope I was fully abolished. However, the reactivity of MAbs recognizing epitope II was not affected. MAbs 31 and 32, recognizing epitopes I and II, respectively, were selected for the cross-reactivity to heterologous reovirus strains. The results suggest that the two epitopes are highly conserved among these virus strains. A MAb capture enzyme-linked immunosorbent assay (ELISA) procedure was developed using MAbs 32 and 31 to detect reovirus protein sigmaA in samples from tendon tissues of infected bird and chicken embryo fibroblast (CEF) cell cultures. Avian reovirus sigmaA antigens in tendon specimens were detected from the inoculated birds as early as 2 days post-inoculation (PI), approximated a peak at 7 days PI, and maintained this until 16 days PI, then decreased gradually. A clear difference in absorbance values between the tendon samples of the avian reovirus- and mock-infected birds is obtained. Positive results were also obtained from avian reovirus-infected CEF and from the tendon tissues of naturally infected broilers. These results indicated that the MAb capture ELISA is a useful methods for the detection of avian reovirus from chickens suspected to have avian reovirus infections.     

Frequent use of colistin-based drug treatment to eliminate extended-spectrum beta-lactamase-producing <Emphasis Type="Italic">Escherichia coli</Emphasis> in backyard chicken farms in Thai Binh Province,Vietnam

Reports of livestock infections with extended-spectrum beta-lactamase-producing Escherichia coli (ESBL-E) are increasing. Based on interviews conducted over a 6-month period, we found that veterinarians in the Vietnamese province of Thai Binh prefer to prescribe colistin-based drugs (CBD) in chicken farms. We aimed to clarify whether CBD use selects for strains of colistin-resistant ESBL-E. With the cooperation of seven local households, we detected ESBL-E in chickens’ feces after treating chickens with CBD. Phylogenetic groupings and the presence of CTX-M/AmpC genes were determined, and the multi-antibiotic susceptibility of isolates was analyzed. Our results showed that ESBL-E presented in seven chickens’ feces from two households. Seventy-two percent of ESBL-E isolates harbored CTX-M9 and the phylogenetic group A; the colistin minimum inhibitory concentration (MIC) of all isolated ESBL-E ranged from 0.064 to 1 μg mL^?1. Moreover, ESBL-E isolates were used to experimentally select for colistin resistance, and the effect of commercial CBD on ESBL-E was investigated. The results showed that an ESBL-E strain with a colistin MIC of 4 μg mL^?1 was able to grow in media with CBD. Although CBD treatment was effective, in vitro experiments demonstrated that ESBL-E can easily acquire colistin resistance. Therefore, restrictions on colistin use are necessary to prevent the emergence of colistin-resistant bacteria.     

不同采毛方式对长毛兔血清蛋白及含氮代谢物水平的影响

文章分析了3种方式（剪毛、直接拔毛和药物拔毛）对皖系长毛兔采毛前、采毛后0、4、8、24、48 h血清蛋白及含氮代谢物变化规律的影响.结果发现：3种采毛方式均显著上调血清总蛋白和肌酐水平,而血清白蛋白仅受药物拔毛影响而显著上调;药物拔毛显著上调血清总胆红素和直接胆红素水平,同时直接胆红素还受其他两种采毛方式影响而显著下调;两种拔毛方式（直接拔毛和药物拔毛）显著上调血清尿素和尿酸水平,但剪毛对这两个指标没有影响.长毛兔在采毛后48 h内,多数血清指标可恢复到采毛前水平,或呈恢复趋势;该研究筛选获得采毛应激指标血清总蛋白和肌酐,拔毛应激指标尿素和尿酸,同时发现肌酐、白蛋白和胆红素受地塞米松的显著影响.该研究结果可为初步揭示不同采毛方式应激生化机制提供参考.     

Inhibition of FGF Signalling Pathway Augments the Expression of Pluripotency and Trophoblast Lineage Marker Genes in Porcine Parthenogenetic Blastocyst

The consistent failure to isolate bona fide pluripotent cell lines from livestock indicates that the underlying mechanisms of early lineage specification are poorly defined. Unlike other species, the contrivances of segregation have been comprehensively studied in the mouse. In mouse, FGF/MAPK signalling pathway dictates the segregation of hypoblast (primitive endoderm). However, it is not evident whether this mechanism is also conserved in livestock. Here, in this study, we examined the roles of FGF/MAP kinase signalling pathways in porcine parthenogenetic embryos during the early development. Porcine parthenogenetic embryos were cultured in the medium addition with FGFR inhibitor BGJ398 (10 μm ) or DEMOS. Pluripotency‐ and lineage‐related gene expressions in the early porcine embryos were determined. Compared to control, total cell numbers on day 7 were significantly higher (55 ± 5.96 vs 47 ± 1.97, p < 0.05) in embryos cultured in the presence of BGJ398, but had no significant effect on the rate of blastocyst development (47% vs 44%, p > 0.05). Nonetheless, BGJ398 treatment significantly augmented the expression of pluripotency and trophoblast marker genes (SOX2, OCT4, KLF4 and CDX2), but did not significantly change the expression of NANOG and hypoblast marker gene (GATA4). Furthermore, the addition of FGF signalling agonist (FGF2) during the embryo development significantly decreased the expression of pluripotency and trophoblast marker genes (SOX2, NANOG, KLF4 and CDX2), but no significant effect on the expression of OCT4 and GATA4 was observed. Here, we exhibit that inhibition of FGF signalling could improve the quality of the porcine embryo and escalate the chance to capture pluripotency. Besides, it also promotes the trophoblast development of porcine parthenogenetic embryo. In addition, the data suggested that FGF signalling pathway is dispensable for the segregation of hypoblast and epiblast lineages in porcine embryo during the early development.     

Aortic rupture causing cardiac tamponade in a 24‐day‐old Friesian colt with concurrent colonic Chlamydiosis and Balantidiasis

A 24‐day‐old Friesian colt died suddenly and a physical examination the morning the foal died showed no abnormalities and serum IgG levels >8.0 g/l. Necropsy examination revealed haemopericardium and a 2 cm transverse tear at the root of the aorta. The foal was also found to have Chlamydophila spp. in the epithelium and Balantidium coli on the mucosal surface of the large colon. An aortic rupture is a novel finding in a foal, colonic Chlamydiosis has not been previously reported in horses and Balantidium coli has not been reported in equids in North America.     

Intake and digestibility in sheep and chemical composition during different seasons of some West African browse species

Foliage of Afzelia africana, Pterocarpus erinaceus and Khaya senegalensis, from 10 trees per species, was collected every two weeks during the late dry, rainy and cool season to determine the seasonal effects on chemical composition. Fifteen rams of the Djallonké breed, weighing on average 20.0 kg, were used to evaluate the voluntary intake and digestibility of hay of A. gayanus, foliage of A. africana (as a sole feed), and A. africana, P. erinaceus and K. senegalensis offered with 30% of the diet as A. gayanus hay. The crude protein (CP) content of A. africana, and P. erinaceus decreased significantly from the late dry season to the cool season when that of K. senegalensis tended to increase. The mean CP of A. africana, P. erinaceus and K. senegalensis differed significantly (173 g, 139 g and 114 g/kg DM, respectively). The DM intake of A. africana offered with hay (571 g/d) or as a sole feed (598 g/d) were not significantly different, but was higher than that of P. erinaceus (428 g/d) and K. senegalensis (298 g/d). The digestibility calculated by difference of DM and CP of A. africana (582 g/kg DM and 795 g/kg CP, respectively) did not differ significantly from A. africana as a sole feed, but were higher than for the other species. The nutritive value of A. africana seems to justify the high preference of herders for this species.     

规模化屠宰场官方兽医及协检员的设置与职责

@@进入新世纪以来,伴随SARS、禽流感和猪链球菌病等疫情在全球范围内的传播,对世界各国人民的身体健康构成极大威胁。因此,国家逐步加大对动物疫病的防控,相继颁布了《中华人民共和国动物防疫法》《动物检疫管理办法》等法律法规,使我国动物防疫和检疫工作走上了有法可依、有章可循的规范化、法制化轨道。屠宰检疫是动物检疫工作的重要组成部分,是保证人民群众食肉安全的重要手段。猪肉及猪产品是广大群众餐桌上不可或缺的食品,国家为保证生猪产品质量安全,保障人民身体健康对生猪屠宰实行定点屠宰、集中检疫,从而使重大疫病的发生和流行得到有效控制。     

Wild ostrich (<Emphasis Type="Italic">Struthio camelus australis</Emphasis>) reproduction in Orbata,a nature reserve in Tunisia

The aim of the current investigation was to determine wild ostrich reproductive behaviour in Orbata Nature Reserve by observing 16 hens and 28 cocks over a seven-year period. Intense laying commenced in January, one month after the cessation of the rainy season, and 92% of the eggs were produced during the dry season (January to May, peaking in March). Over the seven years, 1,322 eggs were laid in 69 nests, which corresponded to an annual average production of 19.2 ± 9.1 eggs/nest and 11.8 eggs/hen. 24 nests (34.78%) were non-brooded, 17 nests (24.64%) were deserted in the course of incubation, and 28 nests (40.58%) possessed hatched eggs. All the non-incubated nests had egg losses equivalent to 46.6 ± 12.6%. Hatchability success of incubated eggs was 41.9 ± 12.0%. Ostriches tended to dig their nests adjacent to the reserve enclosure which had direct access by road and track, the latter subjecting them to human disturbance and predation. The systematic obstruction of these nests stimulated ostriches to build additional nests within the reserve perimeter. The authors discussed the results recorded in an ostrich flock in relation to the environmental factors (climatic factors, food disponibility and predation) and suggested possibilities for improved wildlife management.