Identification of a bisegmented double-stranded RNA virus (picobirnavirus) in calf faeces

To determine the incidence of rotavirus infection among dairy herds in the State of S?o Paulo, Brazil, 576 faecal samples obtained from calves aged 1-45 days with and without diarrhoea, reared on 63 dairy cattle farms, were analyzed. Polyacrylamide gel electrophoresis (PAGE) identified 28 samples positive for group A rotavirus, while four samples, two diarrhoeic and two non-diarrhoeic, showed a bisegmented genome with a typical picobirnavirus pattern. Electron microscopy revealed spherical virus particles with a diameter of 37 nm and without a defined surface structure. The present study is the first report of a bisegmented virus identified in cattle in Brazil.     

Identification of picobirnavirus, viruses with bisegmented double stranded , in rabbit faeces

Picobirnaviruses are a novel group of viruses recently found in the faeces of several species of vertebrates. Examination by polyacrylamide gel electrophoresis of rabbit faecal samples collected in one animal facility revealed the viruses in 23 (11 per cent) of 211 samples. Further analysis by electron microscopy and caesium chloride isopycnic centrifugation confirmed the presence of picobirnaviruses in the samples. The oral inoculation of three newly weaned rabbits with purified virus resulted in the excretion of a virus with an electropherotype similar to the inoculum, by two of the three inoculated animals. Maximal viral shedding was detected 13 days after inoculation. No sign of diarrhoea was observed either in the inoculated animals or in the virus excreting animals surveyed. No antibody activity could be detected in the paired serum samples taken from the inoculated animals.     

Viruses and virus-like particles in the faeces of dogs with and without diarrhoea

Negative staining electron microscopy was used to identify viruses in 157 normal and 29 diarrhoeal faecal samples collected from 156 dogs admitted to an animal shelter during an 8 month period (March to October) in 1982. Seven distinct viral types were detected: 21-26 nm parvovirus-like particles, 28-31 nm astrovirus-like particles, a previously undescribed 34-35 nm "round" virus particle, coronavirus, coronavirus-like particles ( CVLP ), rotavirus and papova-like virus. Parvovirus-like particles alone were detected in 14 diarrhoeal and 50 normal faeces, astrovirus-like particles in 3 normal faeces, "round" viruses in 4 normal faeces, coronavirus in 2 diarrhoeal and 5 normal faeces, CVLP in one diarrhoeal and one normal faeces, rotavirus in 2 normal faeces, papova-like virus in one normal faeces, both parvovirus-like particles and coronavirus in 2 diarrhoeal and 2 normal faeces, parvovirus-like particles and rotavirus in one normal faeces and parvovirus-like and papova-like virus in one normal faeces. The significance of these findings in canine and human disease is discussed.     

负链RNA病毒的反向遗传技术

反向遗传技术是一种新的分子生物学技术,它在深入研究负链RNA病毒基因组结构和功能,探寻其基因组复制、转录和研发新型基因工程疫苗上发挥着重要的作用。文章介绍了分节与不分节负链RNA病毒的复制机理和特征,论述了反向遗传学在研究这些病毒上的策略、最新研究进展及其主要影响拯救效率的因素,进一步涉及了负链RNA病毒载体及其重组病毒的研究动态。因此,通过反向遗传学,人们将更加了解负链RNA病毒,为科学利用和有效控制该病毒奠定基础。     

RNA病毒重组及发生机制

重组在RNA病毒中很常见,但有关重组的许多关键问题目前尚无法解释.本文主要针对RNA重组的发生机制及其影响因素,解释重组频率发生变化的原因,阐述RNA重组未来仍需解决的问题.RNA重组是RNA病毒特殊的遗传信息交换方式,其频率在不同的RNA病毒中显著不同.目前,研究者普遍认为重组发生的机制可分为复制型、非复制型重组以及...     

Prevalence of vancomycin-resistant Enterococcus faecium (VREF) in pig faeces from slaughterhouses in Spain

A study to estimate the prevalence of vancomycin-resistant Enterococcus faecium in faecal samples from pigs at slaughterhouses in Spain was carried out between November 1998 and January 1999 with 900 samples taken from four abattoirs representing 9.7% of all pig slaughtered in 1998. Using a selective enrichment broth with vancomycin (8 μg/ml), 64 samples (7.1%; 95% CI: 5.5, 9.0%) had E. faecium vancomycin-resistant strains that showed minimal inhibitory concentrations of 256 μg/ml (62 strains) and 512 μg/ml (two strains). Results by farm showed that 43 of the 240 pig farms represented in the sampling had at least one faecal sample with vancomycin-resistant E. faecium.     

不分节段负股RNA病毒作为活疫苗载体研究进展

不分节段负股RNA病毒（nonsegmented negative—strand RNA viruses,NNSV）属于单负股病毒目,具有许多优良的特性,可以作为活病毒疫苗的候选载体,利用反向遗传学系统,表达外源基因,研发新型疫苗。NNSV载体疫苗的免疫原性,载体的容量,外源糖蛋白对载体病毒生物学的影响,以及插入基因的遗传稳定性等已成为当前活病毒疫苗载体研发的重要课题。     

Rapid methods for comparing the double-stranded RNA genome profiles of bluetongue virus

Various double-stranded RNA extraction procedures, gel electrophoresis systems, and methods to detect the RNA bands in the gel were investigated to find the most rapid methods to obtain the genome profiles of bluetongue virus in small volumes (1–25 ml) of infected cell culture fluids. Rapid double-stranded RNA extraction procedures coupled with staining the acrylamide gel slabs with ethidium bromide or silver nitrate resulted in well-defined genome profiles from bluetongue virus infected cell cultures in 6–48 h. Radioactive labelling of viral RNA with ³²P was time consuming, cumbersome and expensive. These techniques detect less than 0.5 μg of double-stranded RNA which can be obtained from one 1-ml well of a 24-well cluster plate of bluetongue virus infected cell monolayers. The methods were therefore suitable for rapid comparisons of the electropherotypes of multiple virus isolates.     

Sequence analysis of four double-stranded RNA genomic segments reveals an orthoreovirus with a unique genotype infecting psittaciformes

This paper describes the characterization of four double-stranded ribonucleic acid segments, S1, S2, S3, and S4, of a newly identified pathogenic reovirus from parrots. The four segments share a unique 5' terminus GCUUUUC. The amino-acid sequences of the conserved sigma A and sigma NS proteins show less than 60% sequence similarity, whereas those of the outer capsid proteins sigma B and sigma C have at most 47% sequence similarity to their counterparts in other bird or bat reoviruses. In a phylogenetic analysis of the amino-acid sequences, the proteins coded for by the S1 segment, P10, P17, and sigma C, group with their homologous proteins in other avian reoviruses, whereas the major capsid protein, sigma B, and the nonstructural protein, sigma NS, show more sequence similarity to their bat reoviral counterparts. The phylogenetic relationship of sigma A with the homologous avian and bat sequences is unresolved. The possibility that the parrot reovirus has evolved from an ancestral, more batlike reovirus is discussed. It is proposed to designate this unique virus as PsRV.     

Prevalence of vancomycin-resistant Enterococcus faecium (VREF) in pig faeces from slaughterhouses in Spain

A study to estimate the prevalence of vancomycin-resistant Enterococcus faecium in faecal samples from pigs at slaughterhouses in Spain was carried out between November 1998 and January 1999 with 900 samples taken from four abattoirs representing 9.7% of all pig slaughtered in 1998. Using a selective enrichment broth with vancomycin (8 μg/ml), 64 samples (7.1%; 95% CI: 5.5, 9.0%) had E. faecium vancomycin-resistant strains that showed minimal inhibitory concentrations of 256 μg/ml (62 strains) and 512 μg/ml (two strains). Results by farm showed that 43 of the 240 pig farms represented in the sampling had at least one faecal sample with vancomycin-resistant E. faecium.     

动物RNA病毒反向遗传学操作技术及应用进展

本文简述了反向遗传操作技术的基本原理以及近年来动物RNA病毒基因复制与表达调控机理、病毒与宿主间的相互作用,以及构建新型病毒载体和研发新型疫苗等方面的研究进展。     

Translation of double-stranded RNA segments 1 and 2 of Chuzan virus in rabbit reticulocyte lysates.

Coding assignments of genome segments 1 and 2 of Chuzan virus strain K-47 were studied in vitro by using rabbit reticulocyte lysates. The double-stranded RNA segment was extracted from sodium dodecyl sulfate (SDS)-polyacrylamide gels by a modified crushing and eluting technique. Translation products labeled with [35S]methionine were resolved by SDS-polyacrylamide gel electrophoresis. The molecular weights of the products from RNA segments 1 and 2 were estimated to be 98 and 95 kilodaltons, respectively.     

The critical threshold of Lawsonia intracellularis in pig faeces that causes reduced average daily weight gains in experimentally challenged pigs

Serology indicates that Lawsonia intracellularis infection is widespread in many countries, with most pigs seroconverting before 22 weeks of age. However, the majority of animals appear to be sub-clinically affected, demonstrated by the low reported prevalence of diarrhoea. Production losses caused by sub-clinical proliferative enteropathy (PE) are more difficult to diagnose, indicating the need for a quantitative L. intracellularis assay that correlates well with disease severity. In previous studies, increasing numbers of L. intracellularis in pig faeces, quantified with a real time polymerase chain reaction (qPCR), showed a strong negative correlation with average daily gain (ADG).In this study, the association between faecal L. intracellularis numbers and PE severity was examined in two L. intracellularis experimental challenge trials (n₁ = 32 and n₂ = 95). The number of L. intracellularis shed in individual faeces was determined by qPCR on days 0, 7, 14, 17 and 21 days post challenge, and average daily gain was recorded over the same period. The severity of histopathological lesions of PE was scored at 21 days post challenge. L. intracellularis numbers correlated well with histopathology severity and faecal consistency scores (r = 0.72 and 0.68, respectively), and negatively with ADG (r = ?0.44). Large reductions in ADG (131 g/day) occurred when the number of L. intracellularis shed by experimentally challenged pigs increased from 10⁷ to 10⁸ L. intracellularis, although smaller ADG reductions were also observed (15 g/day) when the number of L. intracellularis increased from 10⁶ to 10⁷ L. intracellularis.