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1.
The open reading frame of the S3 segment encoding the sigma2 protein of four turkey reovirus field isolates was analyzed for sequence heterogeneity. The turkey reoviruses we present here have a 97% amino acid identity to turkey NC 98. The S3 nucleotide and amino acid sequence similarity was < or =61% and 78%-80%, respectively, when compared to the chicken reovirus isolates. Comparison of amino acid sequences from chickens and turkeys with that of a duck isolate revealed a 53% and 55% similarity, respectively. Phylogenetic analyses, based on both nucleotide and amino acid sequence, resulted in three major groups among the avian reoviruses; these groups were clearly separated by species. The results of this study provide further evidence, based on the deduced sigma2 sequence, that turkey reoviruses form a distinct, separate group relative to chicken and duck isolates. In addition, as a result of the limited sequence identity with their avian counterparts, turkey reoviruses could potentially be considered a separate virus species within subgroup 2 of the Orthoreovirus genus.  相似文献   

2.
RNA viruses rapidly mutate, which can result in increased virulence, increased escape from vaccine protection, and false-negative detection results. Targeted detection methods have a limited ability to detect unknown viruses and often provide insufficient data to detect coinfections or identify antigenic variants. Random, deep sequencing is a method that can more fully detect and characterize RNA viruses and is often coupled with molecular techniques or culture methods for viral enrichment. We tested viral culture coupled with third-generation sequencing for the ability to detect and characterize RNA viruses. Cultures of bovine viral diarrhea virus, canine distemper virus (CDV), epizootic hemorrhagic disease virus, infectious bronchitis virus, 2 influenza A viruses, and porcine respiratory and reproductive syndrome virus were sequenced on the MinION platform using a random, reverse primer in a strand-switching reaction, coupled with PCR-based barcoding. Reads were taxonomically classified and used for reference-based sequence building using a stock personal computer. This method accurately detected and identified complete coding sequence genomes with a minimum of 20× coverage depth for all 7 viruses, including a sample containing 2 viruses. Each lineage-typing region had at least 26× coverage depth for all viruses. Furthermore, analyzing the CDV sample through a pipeline devoid of CDV reference sequences modeled the ability of this protocol to detect unknown viruses. Our results show the ability of this technique to detect and characterize dsRNA, negative- and positive-sense ssRNA, and nonsegmented and segmented RNA viruses.  相似文献   

3.
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4.
A new reovirus was isolated from a sick Muscovy duckling with hemorrhagic-necrotic lesions in the liver in Zhejiang, China in 2000 and was tentatively denoted a new type of Muscovy duck reovirus (N-MDRV ZJ00M). This reovirus was propagated in a chicken fibroblast cell line (DF-1) with obvious cytopathic effects. The reovirus's genome was 23,419 bp in length with an approximately 50% G+C content and 10 dsRNA segments encoding 12 proteins. The length of the genomic segments was similar to those of avian reoviruses (ARVs), which range from 3959 nt (L1) to 1191 nt (S4) in size. All of the segments have the conserved terminal sequences 5′-GCUUUUU…UUCAUC-3′, and all of the genome segments, with the exception of S1, apparently encoded one single primary translation product. The genome analysis revealed that the S1 segment of N-MDRV is a tricistronic gene that encodes the overlapping ORFs for p10, p18, and σC. This finding is similar to that found for ARVs but distinct from that found for classical MDRV and GRV, which have a bicistronic S4 segment that encodes p10 and σC and do not encode p18. The amino acid (aa) alignments of the putative proteins encoded by the main ORF in each segment revealed a high similarity (14.1–100%) to the counterpart proteins encoded by other ARV species from the avian orthoreoviruses (e.g., ARV, classical MDRV and N-MDRV) in the Orthoreovirus genus, particularly with N-MDRV (94.6–100%). The phylogenetic analysis of the nucleotide sequences of all 10 genome segments revealed that N-MDRV ZJ00M is distinct from all other described reovirus species groups but is a separated from the ARV (including MDRV and GRV) species within orthoreovirus species group II and grouped into the classical MDRV and GRV genogroup with the N-MDRV isolates. The MDRV genogroup can be further divided into two genotype clusters. The morphological and pathological analyses and the genetic characterization of N-MDRV ZJ00M suggest that it belongs to genotype 2 (N-MDRV). In addition, the RT-PCR assays of DRV diseased duckling and gosling samples collected from different regions of China during 2000–2013 indicate that N-MDRV is currently the prevalent genotype in China.  相似文献   

5.
In 2009, 26 clinical samples (organs and oral/cloacal swabs) from a total of 24 corn snakes (Pantherophis guttatus) from a single owner were sent to our laboratory to be tested for the presence of viruses. Paramyxoviruses (PMV), adenoviruses (AdV) and reoviruses were detected by RT-PCR, PCR and virus isolation methods. Three snakes were infected with all three viruses at the same time, while two other snakes had a double infection (PMV and reo, AdV and reo) and nine other snakes had a single infection with any of the three viruses. No viruses were detected in 10 animals. All isolated reoviruses were identical to one another and to the reptilian orthoreovirus isolate 55-02 in the partial RNA dependent RNA polymerase (RDRP) gene sequence. AdV partial polymerase sequences represented four different types, one of which was first described here: most similar to SnAdV-1, while the other three were identical to known types: SnAV-1, -2 and -3. However, the detected single PMV differed distinctly from described reptile PMV and was a new type. According to partial L gene, HN gene and U gene sequences it may be the first described representative of a third squamatid PMV cluster: "group C" within the proposed reptilian PMV genus "Ferlavirus". Nucleotide identity values for the L gene of the new PMV compared to group A viruses range between 76.5 and 80.3%, and between 80.5 and 81.2% compared to group B viruses. For the HN gene, these values were similar: 78.2-80% (A) and 79.9-80.5% (B) and somewhat lower for the U gene: 72.7-75.4% (A) and 69.7-70% (B). No reports on the prevalence of concurrent viral infection in captive snake populations have been published so far. The possibility of concurrent infection with several different viruses and subsequent consequences for animal health should be kept in mind when testing reptile samples for viruses.  相似文献   

6.
An orthoreovirus was isolated from an Ostrich (Struthio camelus) and rapidly identified as orthoreovirus by the rapid determination of viral RNA sequences (RDV) system and electron microscopy. Phylogenetic analysis of the sigma A protein indicated that the isolate belonged to avian species and was closely related to chicken orthoreovirus strain 138. The results of the present study indicated that an ostrich orthoreovirus is slight different from other chicken orthoreoviruses and provided evidence of diversity among avian orthoreoviruses. To our knowledge, this is the first genetic report of an orthoreovirus isolated from an ostrich.  相似文献   

7.
Enteric viruses in diarrheic turkey poults   总被引：4，自引：0，他引：4  
Thirty-three intestinal samples from 10-to-21-day-old diarrheic turkey poults were examined for the presence of enteric viruses by electron microscopy. Samples originated from 32 flocks in six commercial operations located in six states. Mortality in these flocks ranged from 3 to 15%, and birds from recovered flocks varied greatly in size. Rotavirus-like agents (RVLA) were the most common viruses associated with diarrhea outbreaks in the flocks examined, occurring in five out of six operations. Other viruses detected either singly or in combination, in order of prevalence, were astroviruses, reoviruses, rotaviruses, enteroviruses, and adenoviruses. With the exception of RVLA and rotaviruses, the other viruses were identified solely on the basis of morphology. Salmonellae were isolated from only one of the intestinal samples. By electron microscopy, RVLA were morphologically indistinguishable from rotaviruses, occurring as both 55-nm single-shelled and 70-nm double-shelled particles. However, immune electron microscopy was useful for antigenic differentiation of these two viruses. Turkey rotaviruses reacted with antisera to porcine and bovine rotaviruses, whereas turkey RVLA did not. Neither turkey rotaviruses nor RVLA reacted with antisera to porcine para-rotavirus or an antigenically distinct bovine rotavirus (bovine rotavirus-like agent). Similarly, convalescent anti-turkey RVLA serum (from recovered specific-pathogen-free poults) reacted with homologous virus but did not react with mammalian or avian rotaviruses or reoviruses. Further, RVLA were found to possess RNA electrophoretic migration patterns unlike those of conventional rotaviruses or reoviruses. This trait was used as an additional means of differentiating these viruses.  相似文献   

8.
We have amplified, cloned and sequenced (part of) the open reading frame of the S1 segment encoding the sigma C protein of avian reoviruses isolated from chickens with different disease conditions in Germany and The Netherlands during 1980 up to 2000. These avian reoviruses were analysed phylogenetically and compared with sequences of avian reoviruses in the Genbank database. The avian reoviruses could be grouped in 5 different genotyping clusters and this classification was identical when the sequences were compared of the 5' end, the 3' end or the whole open reading frame of the sigma C protein. Therefore sequencing of either part of the gene encoding the sigma C protein seems to be reliable for classification. We were unable to identify a correlation between sigma C sequences of the avian reoviruses and the disease condition they were isolated from. The sequences found in The Netherlands and in Germany are, like those in Taiwan, more dispersed than the known avian reovirus sigma C sequences in the USA and Australia. We did not establish temporal or geographic differences in the avian reoviruses studied.  相似文献   

9.
The prevalence of reoviruses in commercial chickens with the runting/stunting syndrome, tenosynovitis, and normal chickens was investigated. Reoviruses were isolated from 3-week-old chickens affected with the runting/stunting syndrome and from older chickens with tenosynovitis; viruses were isolated from tissues with and without lesions. Reoviruses were also frequently isolated from rectal contents of normal 3-week-old chickens, and there was serological evidence of previous reovirus infection in all flocks of adult meat breeder chickens examined. The widespread occurrence of reoviruses in both normal and diseased chickens indicates that the isolation of reoviruses from tissues of chickens with lesions does not necessarily imply any aetiological relationship of reovirus with disease, even in the absence of other known causes. However, the occurrence of reovirus in normal chickens does not preclude an aetiological relationship with disease and further investigation of strain variation and possible virulence factors in avian reoviruses is required.  相似文献   

10.
Characterization of avian reovirus strain-specific polymorphisms   总被引：2，自引：0，他引：2  
Avian reoviruses have been associated with several pathologic conditions, but correlative relationships between genotypes and specific diseases have not been demonstrated. Six avian reoviruses (883, 176, 81-5, S1133, FC, and TX) were selected for this study, and a comparative study of the pathogenic properties of the viruses in chickens, following peroral and footpad inoculation, was carried out, along with a comparison of the electrophoretic mobility of viral genomic segments and viral proteins encoded by the gene segments. The pathogenic properties of the viruses were shown to be diverse, with three distinct pathotypes being defined: Pathotype I (883) caused only a syndrome that we have termed "transient digestive system disorder" (TDSD); Pathotype II (FC, TX, and S1133) caused only "viral arthritis syndrome" (VAS), whereas Pathotype III (176 and 81-5) caused both TDSD and VAS. Likewise, the genomes of the viruses were shown to be extremely polymorphic, with a maximum of five segments co-migrating between any two strains. Considerable variation in the electrophoretic mobility of the encoded proteins also was demonstrated with pronounced variation in the molecular size of the sigma 4 protein, the purported viral attachment protein, being evident. These results show that the genomes of avian reoviruses were extremely polymorphic, preventing correlation between genotypes and pathotypes. But these studies have provided us with the genetic elements needed to characterize the gene functions involved in viral pathogenesis.  相似文献   

11.
Necropsy and culture results are presented for 269 consignments of imported birds (mainly psittacine and passerine species) examined between January 1977 and August 1980. Consignments were submitted for diagnosis of clinical illness or deaths occurring among these birds while they were in quarantine before entry into Canada. Enteritis and injury were the most frequent diagnoses. Pathogens or potential pathogens were isolated from 77% of consignments. Newcastle disease virus was isolated nine times, and Chlamydia psittaci was isolated once. Escherichia coli (from 113 consignments) and salmonellae (from 49) were the most common bacteria isolated, and reoviruses (from 22) and paramyxoviruses other than Newcastle disease virus (from 22) were the most common viruses. Salmonella typhimurium was the most common Salmonella serovar. Salmonella hadar was isolated from turkey poults imported from Great Britain. The possible public health significance of the role of imported birds in the introduction of exotic Salmonella serovars, or of serovars resistant to several antimicrobials is discussed.  相似文献   

12.
Specimens from the legs of chickens from 84 outbreaks of suspected tenosynovitis were examined for the presence of viruses by culture in chick embryo lung or liver cell monolayers. All samples were from broilers or broiler breeders, ranging in age from dead-in-shell embryos to 36 weeks old. Twenty-five outbreaks (29.8 per cent) yielded viruses of which 12 were reoviruses alone, 12 adenoviruses alone, and one, a mixture of both types of virus. Rupture of the gastrocnemius tendon was seen in 12 outbreaks and viruses were isolated from six of these: three were reoviruses and three were adenoviruses. Approximately half the affected flocks from which specimens were received were in the six to 14 week age range. With one exception, all the reovirus isolations were made from chickens of 11 weeks or under, while adenovirus isolations showed more scatter with regard to age.  相似文献   

13.
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14.
Many viruses have been identified in pericardial fluid and in tissue samples from humans with pericarditis by means of molecular diagnostics. In canine idiopathic pericardial effusion there is as yet no conclusive evidence to support the involvement of an infectious agent. This study was designed to investigate a possible relationship between idiopathic pericardial effusion in dogs and viruses most commonly encountered in humans affected with viral pericarditis. Coxsackievirus B3 RNA, influenza virus type A RNA, human adenovirus type 2 DNA, human cytomegalovirus DNA, and parvovirus B19 DNA were investigated using PCR on pericardial effusion samples and pericardial tissue specimens collected from 14 dogs with idiopathic pericardial effusion. PCR was also used to test for two bacteria, Borrelia burgdorferi and Chlamydia pneumoniae. The same microorganisms were also looked for in pericardial effusions or pericardial washes from 10 dogs with neoplastic pericardial effusion, and in samples collected from 10 dogs which died of a non-cardiac disease. One pericardial effusion sample from a dog with the idiopathic form of the disease tested positive for influenza virus type A and sequencing of the amplicon confirmed the PCR result. In another dog from the same group a cytomegalovirus was detected by PCR in the effusion, but sequencing showed this to be a false-positive result. The genomes of the microorganisms investigated were not detected in neoplastic effusions or pericardial washes. The results indicate that viral and bacterial DNA/RNA of relevance for human pericarditis is rare in pericardial samples from dogs with idiopathic pericardial effusion. The finding of influenza type A viral RNA in pericardial fluid from one dog with the idiopathic form of the disease warrants further investigation.  相似文献   

15.
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16.
A number of virologic and environmental factors are involved in the emergence and re-emergence of viral disease. Viruses do not conservatively occupy a single and permanent ecological niche. Rather, due to their intrinsic capacity for genetic change, and to the evolvability of fitness levels, viruses display a potential to parasitize alternative host species. Mutation, recombination and genome segment reassortment, and combination of these molecular events, produce complex and phenotypically diverse populations of viruses, which constitute the raw material on which selection acts. The majority of emerging viral diseases of humans have a zoonotic origin. Sociologic and ecologic factors produce diverse and changing environments in which viral subpopulations have ample opportunities to be selected from intrinsically heterogeneous viral populations, particularly in the case of RNA viruses. In this manner, new human, animal and plant viruses have emerged periodically and, from all evidence, will continue to emerge. This article reviews some of the mechanisms that have been identified in viral emergence, with a focus on the importance of genetic variation of viruses, and on the general concept of biological complexity.  相似文献   

17.
M R Rekik  A Silim 《Avian diseases》1992,36(2):237-246
Total RNA of eight avian reovirus isolates and the S1133 strain were compared by RNase T1-oligonucleotide mapping. The viruses were propagated in Vero cell cultures, and viral genomes were extracted from purified virions for comparison. Pairwise comparisons of the oligonucleotide maps showed genetic variation among reovirus isolates ranging from 78% to 99%. The T1 fingerprints of the RNA of isolates 1103, 724, 615, and 684 differed slightly from the standard S1133 strain, suggesting that the vaccine strain might have changed and became part of the circulating reoviruses. In contrast, when compared with the vaccine strain, isolates 902, 644, and 6207 showed greater differences in the fingerprint pattern. This genomic diversity may be due to the differences in immunological status of the affected avian population and/or due to simultaneous coinfection with different reovirus strains.  相似文献   

18.
A group of avian reoviruses comprising serially passaged S1133 strains and their vaccine derivatives was examined biochemically to study the temporal evolution of the viruses and biologically to assess their relative pathogenicities. The strains fell into three groups of differing virulence, the viruses becoming less pathogenic the longer they were passaged. Protein and RNA profiles of the strains showed no distinct patterns of evolution nor any trend that could be correlated with pathogenicity. Nucleic acid hybridization studies of the strains indicated that all the genes were altered to some extent during passage. The S1 and M3 genes appeared to change the most during the first half of passage history, but later, as the virus was cold-adapted or passaged extensively, the M2, S2, and S3 genes also appeared to vary. When viruses were grouped according to virulence, the greatest changes were seen in the S1, M2, and M3 genes, suggesting that these may be associated with the virulence of a given avian reovirus strain.  相似文献   

19.
Several avian reoviruses were screened to determine their effects on the immune system by inoculating them subcutaneously (SQ) into day-old chicks. For comparison, infectious bursal disease virus (IBDV) was similarly evaluated. The response of the immune system was measured functionally by the hemagglutination-inhibition (HI) response to Newcastle disease virus (NDV) and structurally by changes in the organ-to-body-weight ratios of the bursa of Fabricius, thymus, and spleen. When inoculated SQ, most of the reoviruses caused transient alterations in lymphoid organ weights, decreasing the bursa weight and increasing the spleen weight. Of those reoviruses tested, only one--a commercial vaccine based on the isolate S-1133--demonstrated the ability to interfere significantly with NDV-HI responses, although several had numerically lower titers. Two of the isolates were also evaluated by oral inoculation. Giving the viruses orally did not cause any alterations in organ weights; however, both isolates depressed the HI response of chicks to NDV. Compared with reoviruses, IBDV significantly depressed NDV-HI titers. The structural responses to IBDV differed, however: IBDV significantly depressed bursa weights for all 3 weeks of the test period without affecting spleen weights. Some of the reovirus isolates inoculated SQ were lethal for day-old chicks. This, and their ability to alter the lymphoid-to-body-weight ratios of the spleen and bursa, could be considered valid criteria by which to study the pathogenesis of these agents.  相似文献   

20.
In this study, we isolated and identified three camel pox viruses (CMLV) from two outbreaks of camel pox infection in camels associated with eruptions on cheeks, nostrils, limbs, scrotum, and sheath that occurred at different places of Bikaner district, Rajasthan (India). The scab specimens collected were subjected for virus isolation in Vero cell culture, and the isolated viruses were characterized by employing polymerase chain reaction (PCR) and sequencing. The causative agent was identified as CMLV, based on A-type inclusion, B5R and C18L genes-specific PCRs and partial sequencing of these genes, which clearly confirmed that the outbreaks were caused by CMLV and identity of CMLV isolates. Further, phylogenetic analysis of partial C18L gene sequences have showed that Indian CMLV are clustered together with other reported isolates/strains.  相似文献   

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