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1

Comparison of the minor coat protein gene sequences of aphid-transmissible and -nontransmissible isolates of Citrus tristeza virus  

Microsoft Academic Search

The nucleotide sequences for the minor coat protein (CPm) gene and its deduced amino acid sequences for two aphid-transmissible\\u000a and two nontransmissible isolates of Citrus tristeza virus (CTV) from symptomless orchard trees of Miyagawa satsuma [Citrus unshiu (Macf.) Marc.] on trifoliate orange [Poncirus trifoliate (L.) Raf.] and declining Washington navel [C. sinensis (L.) Osb.] trees on sour orange (C. aurantium

Ali Barzegar; Heshmat Rahimian; Haleh Hashemi Sohi

2010-01-01

2

The First Identified Citrus tristeza virus Isolate of Turkey Contains a Mixture of Mild and Severe Strains  

PubMed Central

The presence of Citrus tristeza virus (CTV) has previously been reported in citrus growing regions of Turkey. All serologically and biologically characterized isolates including I?d?r, which was the first identified CTV isolates from Turkey, were considered mild isolates. In this study, molecular characteristics of the I?d?r isolate were determined by different methods. Analysis of the I?d?r isolate by western blot and BD-RT-PCR assays showed the presence of MCA13 epitope, predominantly found in severe isolates, in the I?d?r isolate revealing that it contains a severe component. For further characterization, the coat protein (CP) and the RNA-dependent RNA polymerase (RdRp) genes representing the 3? and 5? half of CTV genome, respectively, were amplified from dsRNA by RT-PCR. Both genes were cloned separately and two clones for each gene were sequenced. Comparisons of nucleotide and deduced amino acid sequences showed that while two CP gene sequences were identical, two RdRp clones showed only 90% and 91% sequence identity in their nucleotide and amino acid sequences, respectively, suggesting a mixed infection with different strains. Phylogenetic analyses of the CP and RdRp genes of I?d?r isolate with previously characterized CTV isolates from different citrus growing regions showed that the CP gene was clustered with NZRB-TH30, a resistance breaking isolate from New Zealand, clearly showing the presence of severe component. Furthermore, two different clones of the RdRp gene were clustered separately with different CTV isolates with a diverse biological activity. While the RdRp-1 was clustered with T30 and T385, two well-characterized mild isolates from Florida and Spain, respectively, the RdRp-2 was most closely related to NZRB-G90 and NZRB-TH30, two well-characterized resistance breaking and stem pitting (SP) isolates from New Zealand confirming the mixed infection. These results clearly demonstrated that the I?d?r isolate, which was previously described as biologically a mild isolate, actually contains a mixture of mild and severe strains. PMID:25288926

Cevik, Bayram; Yardimci, Nejla; Korkmaz, Savas

2013-01-01

3

Citrus tristeza virus-host interactions  

PubMed Central

Citrus tristeza virus (CTV) is a phloem-limited virus whose natural host range is restricted to citrus and related species. Although the virus has killed millions of trees, almost destroying whole industries, and continually limits production in many citrus growing areas, most isolates are mild or symptomless in most of their host range. There is little understanding of how the virus causes severe disease in some citrus and none in others. Movement and distribution of CTV differs considerably from that of well-studied viruses of herbaceous plants where movement occurs largely through adjacent cells. In contrast, CTV systemically infects plants mainly by long-distance movement with only limited cell-to-cell movement. The virus is transported through sieve elements and occasionally enters an adjacent companion or phloem parenchyma cell where virus replication occurs. In some plants this is followed by cell-to-cell movement into only a small cluster of adjacent cells, while in others there is no cell-to-cell movement. Different proportions of cells adjacent to sieve elements become infected in different plant species. This appears to be related to how well viral gene products interact with specific hosts. CTV has three genes (p33, p18, and p13) that are not necessary for infection of most of its hosts, but are needed in different combinations for infection of certain citrus species. These genes apparently were acquired by the virus to extend its host range. Some specific viral gene products have been implicated in symptom induction. Remarkably, the deletion of these genes from the virus genome can induce large increases in stem pitting (SP) symptoms. The p23 gene, which is a suppressor of RNA silencing and a regulator of viral RNA synthesis, has been shown to be the cause of seedling yellows (SY) symptoms in sour orange. Most isolates of CTV in nature are populations of different strains of CTV. The next frontier of CTV biology is the understanding how the virus variants in those mixtures interact with each other and cause diseases. PMID:23717303

Dawson, W. O.; Garnsey, S. M.; Tatineni, S.; Folimonova, S. Y.; Harper, S. J.; Gowda, S.

2013-01-01

4

Citrus tristeza virus: characterization of Texas isolates, studies on aphid transmission and pathogen-derived control strategies  

E-print Network

Texas CTV isolates. The majority of the CTV isolates tested contained the most severe CTV types known. In Florida, T. citricida were fed on crude CTV preparations in vitro and could transmit CTV to virus-free receptor plants with two CTV isolates...

Herron, Caroline Mary

2004-11-15

5

Emergence and Phylodynamics of Citrus tristeza virus in Sicily, Italy  

PubMed Central

Citrus tristeza virus (CTV) outbreaks were detected in Sicily island, Italy for the first time in 2002. To gain insight into the evolutionary forces driving the emergence and phylogeography of these CTV populations, we determined and analyzed the nucleotide sequences of the p20 gene from 108 CTV isolates collected from 2002 to 2009. Bayesian phylogenetic analysis revealed that mild and severe CTV isolates belonging to five different clades (lineages) were introduced in Sicily in 2002. Phylogeographic analysis showed that four lineages co-circulated in the main citrus growing area located in Eastern Sicily. However, only one lineage (composed of mild isolates) spread to distant areas of Sicily and was detected after 2007. No correlation was found between genetic variation and citrus host, indicating that citrus cultivars did not exert differential selective pressures on the virus. The genetic variation of CTV was not structured according to geographical location or sampling time, likely due to the multiple introduction events and a complex migration pattern with intense co- and re-circulation of different lineages in the same area. The phylogenetic structure, statistical tests of neutrality and comparison of synonymous and nonsynonymous substitution rates suggest that weak negative selection and genetic drift following a rapid expansion may be the main causes of the CTV variability observed today in Sicily. Nonetheless, three adjacent amino acids at the p20 N-terminal region were found to be under positive selection, likely resulting from adaptation events. PMID:23818960

Davino, Salvatore; Willemsen, Anouk; Panno, Stefano; Davino, Mario; Catara, Antonino; Elena, Santiago F.; Rubio, Luis

2013-01-01

6

Citrus tristeza virus: Evolution of Complex and Varied Genotypic Groups  

PubMed Central

Amongst the Closteroviridae, Citrus tristeza virus (CTV) is almost unique in possessing a number of distinct and characterized strains, isolates of which produce a wide range of phenotype combinations among its different hosts. There is little understanding to connect genotypes to phenotypes, and to complicate matters more, these genotypes are found throughout the world as members of mixed populations within a single host plant. There is essentially no understanding of how combinations of genotypes affect symptom expression and disease severity. We know little about the evolution of the genotypes that have been characterized to date, little about the biological role of their diversity and particularly, about the effects of recombination. Additionally, genotype grouping has not been standardized. In this study we utilized an extensive array of CTV genomic information to classify the major genotypes, and to determine the major evolutionary processes that led to their formation and subsequent retention. Our analyses suggest that three major processes act on these genotypes: (1) ancestral diversification of the major CTV lineages, followed by (2) conservation and co-evolution of the major functional domains within, though not between CTV genotypes, and (3) extensive recombination between lineages that have given rise to new genotypes that have subsequently been retained within the global population. The effects of genotype diversity and host-interaction are discussed, as is a proposal for standardizing the classification of existing and novel CTV genotypes. PMID:23630519

Harper, S. J.

2013-01-01

7

Differential tropism in roots and shoots infected by Citrus tristeza virus.  

PubMed

Virus tropism is a result of interactions between virus, host and vector species, and determines the fate of an infection. In this study, we examined the infection process of Citrus tristeza virus (CTV) in susceptible and resistant species, and found that the tropism of CTV is not simply phloem limited, but tissue specific. In resistant species, virus infection was not prevented, but mostly restricted to the roots. This phenomenon was also observed after partial replacement of genes of one CTV strain from another, despite both parental strains being capable of systemic infection. Finally, the roots remained susceptible in the absence of viral gene products needed for systemic infection of shoots. Our results suggest that all phloem cells within a plant are not equally susceptible and that changes in host or virus may produce a novel tropism: restriction by the host to a location where further virus spread is prevented. PMID:25010274

Harper, S J; Cowell, S J; Robertson, C J; Dawson, W O

2014-07-01

8

Citrus tristeza virus p23: a unique protein mediating key virus-host interactions  

PubMed Central

The large RNA genome of Citrus tristeza virus (CTV; ca. 20 kb) contains 12 open reading frames, with the 3?-terminal one corresponding to a protein of 209 amino acids (p23) that is expressed from an abundant subgenomic RNA. p23, an RNA-binding protein with a putative zinc-finger domain and some basic motifs, is unique to CTV because no homologs have been found in other closteroviruses, including the type species of the genus Beet yellows virus (despite both viruses having many homologous genes). Consequently, p23 might have evolved for the specific interaction of CTV with its citrus hosts. From a functional perspective p23 has been involved in many roles: (i) regulation of the asymmetrical accumulation of CTV RNA strands, (ii) induction of the seedling yellows syndrome in sour orange and grapefruit, (iii) intracellular suppression of RNA silencing, (iv) elicitation of CTV-like symptoms when expressed ectopically as a transgene in several Citrus spp., and (v) enhancement of systemic infection (and virus accumulation) in sour orange and CTV release from the phloem in p23-expressing transgenic sweet and sour orange. Moreover, transformation of Mexican lime with intron-hairpin constructs designed for the co-inactivation of p23 and the two other CTV silencing suppressors results in complete resistance against the homologous virus. From a cellular point of view, recent data indicate that p23 accumulates preferentially in the nucleolus, being the first closterovirus protein with such a subcellular localization, as well as in plasmodesmata. These major accumulation sites most likely determine some of the functional roles of p23. PMID:23653624

Flores, Ricardo; Ruiz-Ruiz, Susana; Soler, Nuria; Sanchez-Navarro, Jesus; Fagoaga, Carmen; Lopez, Carmelo; Navarro, Luis; Moreno, Pedro; Pena, Leandro

2013-01-01

9

Effects of Modification of the Transcription Initiation Site Context on Citrus Tristeza Virus Subgenomic RNA Synthesis†  

PubMed Central

Citrus tristeza virus (CTV), a member of the Closteroviridae, has a positive-sense RNA genome of about 20 kb organized into 12 open reading frames (ORFs). The last 10 ORFs are expressed through 3?-coterminal subgenomic RNAs (sgRNAs) regulated in both amounts and timing. Additionally, relatively large amounts of complementary sgRNAs are produced. We have been unable to determine whether these sgRNAs are produced by internal promotion from the full-length template minus strand or by transcription from the minus-stranded sgRNAs. Understanding the regulation of 10 sgRNAs is a conceptual challenge. In analyzing commonalities of a replicase complex in producing so many sgRNAs, we examined initiating nucleotides of the sgRNAs. We mapped the 5? termini of intermediate- (CP and p13) and low- (p18) produced sgRNAs that, like the two highly abundant sgRNAs (p20 and p23) previously mapped, all initiate with an adenylate. We then examined modifications of the initiation site, which has been shown to be useful in defining mechanisms of sgRNA synthesis. Surprisingly, mutation of the initiating nucleotide of the CTV sgRNAs did not prevent sgRNA accumulation. Based on our results, the CTV replication complex appears to initiate sgRNA synthesis with purines, preferably with adenylates, and is able to initiate synthesis using a nucleotide a few positions 5? or 3? of the native initiation nucleotide. Furthermore, the context of the initiation site appears to be a regulatory mechanism for levels of sgRNA production. These data do not support either of the established mechanisms for synthesis of sgRNAs, suggesting that CTV sgRNA production utilizes a different mechanism. PMID:12915539

Ayllon, Maria A.; Gowda, Siddarame; Satyanarayana, Tatineni; Karasev, Alexander V.; Adkins, Scott; Mawassi, Munir; Guerri, Jose; Moreno, Pedro; Dawson, William O.

2003-01-01

10

A genetic system for Citrus Tristeza Virus using the non-natural host Nicotiana benthamiana: an update.  

PubMed

In nature Citrus tristeza virus (CTV), genus Closterovirus, infects only the phloem cells of species of Citrus and related genera. Finding that the CTV T36 strain replicated in Nicotiana benthamiana (NB) protoplasts and produced normal virions allowed development of the first genetic system based on protoplast transfection with RNA transcribed from a full-genome cDNA clone, a laborious and uncertain system requiring several months for each experiment. We developed a more efficient system based on agroinfiltration of NB leaves with CTV-T36-based binary plasmids, which caused systemic infection in this non-natural host within a few weeks yielding in the upper leaves enough CTV virions to readily infect citrus by slash inoculation. Stem agroinoculation of citrus and NB plants with oncogenic strains of Agrobacterium tumefaciens carrying a CTV-T36 binary vector with a GUS marker, induced GUS positive galls in both species. However, while most NB tumors were CTV positive and many plants became systemically infected, no coat protein or viral RNA was detected in citrus tumors, even though CTV cDNA was readily detected by PCR in the same galls. This finding suggests (1) strong silencing or CTV RNA processing in transformed cells impairing infection progress, and (2) the need for using NB as an intermediate host in the genetic system. To maintain CTV-T36 in NB or assay other CTV genotypes in this host, we also tried to graft-transmit the virus from infected to healthy NB, or to mechanically inoculate NB leaves with virion extracts. While these trials were mostly unsuccessful on non-treated NB plants, agroinfiltration with silencing suppressors enabled for the first time infecting NB plants by side-grafting and by mechanical inoculation with virions, indicating that previous failure to infect NB was likely due to virus silencing in early infection steps. Using NB as a CTV host provides new possibilities to study virus-host interactions with a simple and reliable system. PMID:23847598

Ambrós, Silvia; Ruiz-Ruiz, Susana; Peńa, Leandro; Moreno, Pedro

2013-01-01

11

Influenza virus isolation.  

PubMed

The isolation of influenza viruses is important for the diagnosis of respiratory diseases in lower animals and humans, for the detection of the infecting agent in surveillance programs, and is an essential element in the development and production of vaccine. Since influenza is caused by a zoonotic virus it is necessary to do surveillance in the reservoir species (aquatic waterfowls), intermediate hosts (quails, pigs), and in affected mammals including humans. Two of the hemagglutinin (HA) subtypes of influenza A viruses (H5 and H7) can evolve into highly pathogenic (HP) strains for gallinaceous poultry; some HP H5 and H7 strains cause lethal infection of humans. In waterfowls, low pathogenic avian influenza (LPAI) isolates are obtained primarily from the cloaca (or feces); in domestic poultry, the virus is more often recovered from the respiratory tract than from cloacal samples; in mammals, the virus is most often isolated from the respiratory tract, and in cases of high pathogenic avian influenza (HPAI) from the blood and internal organs of infected birds. Virus isolation procedures are performed by inoculation of clinical specimens into embryonated eggs (primarily chicken eggs) or onto a variety of primary or continuous tissue culture systems. Successful isolation of influenza virus depends on the quality of the sample and matching the appropriate culture method to the sample type. PMID:22528151

Krauss, Scott; Walker, David; Webster, Robert G

2012-01-01

12

Increase and Patterns of Spread of Citrus Tristeza Virus Infections in Costa Rica and the Dominican Republic in the Presence of the Brown Citrus Aphid, Toxoptera citricida.  

PubMed

ABSTRACT Citrus tristeza virus (CTV) was monitored for 4 years by monoclonal antibody probes via enzyme-linked immunosorbent assay in four citrus orchards in northern Costa Rica and four orchards in the Dominican Republic following the introduction of the brown citrus aphid, Toxoptera citricida. The Gompertz nonlinear model was selected as the most appropriate in most cases to describe temporal increase of CTV. Ordinary runs analysis for association of CTV-positive trees failed to show a spatial relationship of virus status among immediately adjacent trees within or across rows. The beta-binomial index of dispersion for various quadrat sizes suggested aggregations of CTV-positive trees for all plots within the quadrat sizes tested. Spatial autocorrelation analysis of proximity patterns suggested that aggregation often existed among quadrats of various sizes up to four lag distances; however, significant lag positions discontinuous from the main proximity pattern were rare. Some asymmetry was also detected for some spatial autocorrelation proximity patterns. These results were interpreted to mean that, although CTV-positive trees did not often influence immediately adjacent trees, virus transmission was common within a local area of influence that extended two to eight trees in all directions. Where asymmetry was indicated, this area of influence was somewhat elliptical. The spatial and temporal analyses gave some insight into possible underlying processes of CTV spread in the presence of T. citricida and suggested CTV spread was predominantly to trees within a local area. Patterns of longer-distance spread were not detected within the confines of the plot sizes tested. Longer-distance spread probably exists, but may well be of a complexity beyond the detection ability of the spatial analysis methods employed, or perhaps is on a scale larger than the dimensions of the plots studied. PMID:18944934

Gottwald, T R; Garnsey, S M; Borbón, J

1998-07-01

13

Citrus tristeza virus-based RNAi in citrus plants induces gene silencing in Diaphorina citri, a phloem-sap sucking insect vector of citrus greening disease (Huanglongbing).  

PubMed

A transient expression vector based on Citrus tristeza virus (CTV) is unusually stable. Because of its stability it is being considered for use in the field to control Huanglongbing (HLB), which is caused by Candidatus Liberibacter asiaticus (CLas) and vectored by Asian citrus psyllid, Diaphorina citri. In the absence of effective control strategies for CLas, emphasis has been on control of D. citri. Coincident cohabitation in phloem tissue by CLas, D. citri and CTV was exploited to develop a novel method to mitigate HLB through RNA interference (RNAi). Since CTV has three RNA silencing suppressors, it was not known if CTV-based vector could induce RNAi in citrus. Yet, expression of sequences targeting citrus phytoene desaturase gene by CTV-RNAi resulted in photo-bleaching phenotype. CTV-RNAi vector, engineered with truncated abnormal wing disc (Awd) gene of D. citri, induced altered Awd expression when silencing triggers ingested by feeding D. citri nymphs. Decreased Awd in nymphs resulted in malformed-wing phenotype in adults and increased adult mortality. This impaired ability of D. citri to fly would potentially limit the successful vectoring of CLas bacteria between citrus trees in the grove. CTV-RNAi vector would be relevant for fast-track screening of candidate sequences for RNAi-mediated pest control. PMID:24572372

Hajeri, Subhas; Killiny, Nabil; El-Mohtar, Choaa; Dawson, William O; Gowda, Siddarame

2014-04-20

14

Oropouche virus isolation, southeast Brazil.  

PubMed

An Oropouche virus strain was isolated from a novel host (Callithrix sp.) in Arinos, Minas Gerais State, southeastern Brazil. The virus was identified by complement fixation test and confirmed by reverse transcription-polymerase chain reaction. Phylogenetic analysis identified this strain as a genotype III isolate previously recognized only in Panama. PMID:16318707

Nunes, Márcio Roberto Teixeira; Martins, Lívia Carício; Rodrigues, Sueli Guerreiro; Chiang, Jannifer Oliveira; Azevedo, Raimunda do Socorro da Silva; da Rosa, Amelia P A Travassos; Vasconcelos, Pedro Fernando da Costa

2005-10-01

15

Chlorella viruses isolated in China  

SciTech Connect

Plaque-forming viruses of the unicellular, eukaryotic, exsymbiotic, Chlorella-like green algae strain NC64A, which are common in the United States, were also present in fresh water collected in the People's Republic of China. Seven of the Chinese viruses were examined in detail and compared with the Chlorella viruses previously isolated in the United States. Like the American viruses, the Chinese viruses were large polyhedra and sensitive to chloroform. They contained numerous structural proteins and large double-stranded DNA genomes of at least 300 kilobase pairs. Each of the DNAs from the Chinese viruses contained 5-methyldeoxycytosine, which varied from 12.6 to 46.7% of the deoxycytosine, and N{sup 6}-methyldeoxyadenosine, which varied from 2.2 to 28.3% of the deoxyadenosine. Four of the Chinese virus DNAs hybridized extensively with {sup 32}P-labeled DNA from the American virus PBCV-1, and three hybridized poorly.

Zhang, Y.; Burbank, D.E.; Van Etten, J.L. (Univ. of Nebraska, Lincoln (USA))

1988-09-01

16

Mild strain cross protection of tristeza: a review of research to protect against decline on sour orange in Florida  

PubMed Central

Tristeza, caused by Citrus tristeza virus (CTV), has long been present in Florida but outbreaks of decline on sour orange rootstock were occasional events until the late 1970s. Sour orange rootstock was valued for the high quality of fruit produced and was widely used because of its tolerance of citrus blight, a disease of unknown etiology. Research was directed towards the selection and screening of mild strains of CTV which could protect against sour orange decline strains. Following the introduction of Toxoptera citricida (also known as the brown citrus aphid) in 1995 there was a greater concern for maintaining production of existing blocks of citrus on sour orange rootstock. Availability of the CTV genome sequence around the same time as well as molecular characterization of in planta CTV populations led to the selection of mild CTV isolates which when inoculated into existing field trees, extended the productive life of the groves and enabled a more graduate replanting of trees on CTV-tolerant rootstocks. The history of CTV in Florida and the methods developed to select mild isolates for use for mild strain cross protection will be reviewed. PMID:24046764

Lee, Richard F.; Keremane, Manjunath L.

2013-01-01

17

Comparison of Immunohistochemistry and Virus Isolation for Diagnosis of West Nile Virus  

PubMed Central

Immunohistochemistry and virus isolation were performed on 1,057 birds. Immunohistochemistry, virus isolation, or both found 325 birds to be West Nile virus positive. Of these, 271 were positive by both methods. These results indicate that virus isolation and immunohistochemistry are approximately equal in their ability to detect West Nile virus. PMID:15956415

Ellis, Angela E.; Mead, Daniel G.; Allison, Andrew B.; Gibbs, Samantha E. J.; Gottdenker, Nicole L.; Stallknecht, David E.; Howerth, Elizabeth W.

2005-01-01

18

Comparison of cowpox-like viruses isolated from European zoos  

Microsoft Academic Search

Summary Poxviruses isolated from captive carnivores in Russia (Moscow virus) and elephants in Germany (elephant virus) were very closely-related to cowpox virus. Immunological analysis with absorbed sera separated elephant virus but not cowpox and Moscow virus, whereas polypeptide analysis separated cowpox but not elephant and Moscow virus. A combination of biological tests separated all three. The epidemiological implications are briefly

D. Baxby; W. B. Shackleton; Jean Wheeler; A. Turner

1979-01-01

19

Isolation of Usutu Virus in Germany  

PubMed Central

Usutu virus (USUV) is a mosquito-borne flavivirus that emerged 2001 in Austria and caused deaths in wild birds. In Germany, 70,378 female mosquitoes were captured in 2009 and 2010 and assayed for USUV. Virus was isolated in cell culture from one pool of Culex pipiens pipiens mosquitoes trapped exclusively in August 2010 in Weinheim, Germany. Subsequent phylogenetic analysis demonstrated a close relationship between the isolated USUV strain from Germany and a USUV strain from Austria, which was detected in a dead blackbird in 2004. PMID:21896821

Jost, Hanna; Bialonski, Alexandra; Maus, Deborah; Sambri, Vittorio; Eiden, Martin; Groschup, Martin H.; Gunther, Stephan; Becker, Norbert; Schmidt-Chanasit, Jonas

2011-01-01

20

ISOLATION OF NEWCASTLE DISEASE VIRUS FROM TEALS  

Microsoft Academic Search

Eight of 30 teals (Anas crecca) died several days following capture and Newcastle Disease Virus (NDV) was isolated from all eight. Brains from the dead birds were homogenized and inoculated into chicken embryos. The allantoic fluid from the embryos was inoculated into 10 domestic chickens susceptible to NDV and 10 chickens immunized against NDV. Eight of 10 (80%) susceptible chickens

M. H. BOZORGMEHRI-FARD; H. KEYVANFAR

21

Molecular and Epidemiologic Analysis of Dengue Virus Isolates from Somalia  

Microsoft Academic Search

Nucleotide sequence analysis was performed on 14 dengue virus isolates (13 dengue-2 viruses and 1 dengue-3 virus) recovered from febrile soldiers in Somalia in 1993. The dengue-2 viruses were most closely related to dengue-2 virus recovered in Somalia in 1984. However, differences in nucleotide sequence (0.35% to 1.35%) were evident among the 1993 isolates. These differences were closely associated with

Niranjan Kanesa-thasan; Gwong-Jen J. Chang; Bonnie L. Smoak; Alan Magill; M. Jeanne Burrous; Charles H. Hoke

1998-01-01

22

Influenza A virus isolation, culture and identification.  

PubMed

Influenza A viruses (IAVs) cause epidemics and pandemics that result in considerable financial burden and loss of human life. To manage annual IAV epidemics and prepare for future pandemics, an improved understanding of how IAVs emerge, transmit, cause disease and acquire pandemic potential is urgently needed. Fundamental techniques essential for procuring such knowledge are IAV isolation and culture from experimental and surveillance samples. Here we present a detailed protocol for IAV sample collection and processing, amplification in chicken eggs or mammalian cells, and identification from samples containing unknown pathogens. This protocol is robust, and it allows for the generation of virus cultures that can be used for downstream analyses. Once experimental or surveillance samples are obtained, virus cultures can be generated and the presence of IAVs can be verified in 3-5 d via reverse-transcription (RT)-PCR or hemagglutination assay. Increased time frames may be required for less experienced laboratory personnel, or when large numbers of samples will be processed. PMID:25321410

Eisfeld, Amie J; Neumann, Gabriele; Kawaoka, Yoshihiro

2014-11-01

23

High sequence conservation among cucumber mosaic virus isolates from Lily  

Microsoft Academic Search

Summary.  ?For classification of Cucumber mosaic virus (CMV) isolates from ornamental crops of different geographical areas, these were characterized by comparing the nucleotide\\u000a sequences of RNAs 4 and the encoded coat proteins. Within the ornamental-infecting CMV viruses both subgroups were represented.\\u000a CMV isolates of Alstroemeria and crocus were classified as subgroup II isolates, whereas 8 other isolates, from lily, gladiolus, amaranthus,

Y. K. Chen; A. F. L. M. Derks; S. Langeveld; R. Goldbach; M. Prins

2001-01-01

24

Virus Isolation and Identification in Cervical Cancer Patients.  

National Technical Information Service (NTIS)

7 strains of viruses were isolated from cervical specimens of cervical cancer patients. The viral isolates were herpes simplex viruses according to the morphologic, biologic as well as serologic characteristics. 5 of the 7 strains were typed and 3 were he...

J. Xiang, J. Feng, P. Huang, W. Chen, J. Wu

1982-01-01

25

Isolation of Genetically Diverse Marburg Viruses from Egyptian Fruit Bats  

PubMed Central

In July and September 2007, miners working in Kitaka Cave, Uganda, were diagnosed with Marburg hemorrhagic fever. The likely source of infection in the cave was Egyptian fruit bats (Rousettus aegyptiacus) based on detection of Marburg virus RNA in 31/611 (5.1%) bats, virus-specific antibody in bat sera, and isolation of genetically diverse virus from bat tissues. The virus isolates were collected nine months apart, demonstrating long-term virus circulation. The bat colony was estimated to be over 100,000 animals using mark and re-capture methods, predicting the presence of over 5,000 virus-infected bats. The genetically diverse virus genome sequences from bats and miners closely matched. These data indicate common Egyptian fruit bats can represent a major natural reservoir and source of Marburg virus with potential for spillover into humans. PMID:19649327

Towner, Jonathan S.; Amman, Brian R.; Sealy, Tara K.; Carroll, Serena A. Reeder; Comer, James A.; Kemp, Alan; Swanepoel, Robert; Paddock, Christopher D.; Balinandi, Stephen; Khristova, Marina L.; Formenty, Pierre B. H.; Albarino, Cesar G.; Miller, David M.; Reed, Zachary D.; Kayiwa, John T.; Mills, James N.; Cannon, Deborah L.; Greer, Patricia W.; Byaruhanga, Emmanuel; Farnon, Eileen C.; Atimnedi, Patrick; Okware, Samuel; Katongole-Mbidde, Edward; Downing, Robert; Tappero, Jordan W.; Zaki, Sherif R.; Ksiazek, Thomas G.; Nichol, Stuart T.; Rollin, Pierre E.

2009-01-01

26

Distinct replicative and cytopathic characteristics of human immunodeficiency virus isolates.  

PubMed Central

According to their capacity to replicate in vitro, human immunodeficiency virus (HIV) isolates can be divided into two major groups, rapid/high and slow/low. Rapid/high viruses can easily be transmitted to a variety of cell lines of T-lymphoid (CEM, H9, and Jurkat) and monocytoid (U937) origin. In contrast, slow/low viruses replicate transiently, if at all, in these cell lines. Except for a few isolates, the great majority of slow/low viruses replicate in peripheral blood mononuclear cells and Jurkat-tatIII cells constitutively expressing the tatIII gene of HIV-1. The viruses able to replicate efficiently cause syncytium formation and are regularly isolated from immunodeficient patients. Poorly replicating HIV isolates, often obtained from individuals with no or mild disease, show syncytium formation and single-cell killing simultaneously or, with some isolates, cell killing only. Images PMID:2459416

Fenyo, E M; Morfeldt-Manson, L; Chiodi, F; Lind, B; von Gegerfelt, A; Albert, J; Olausson, E; Asjo, B

1988-01-01

27

Characterisation of New Zealand isolates of infectious bursal disease virus  

Microsoft Academic Search

Summary.  ?Isolates of infectious bursal disease virus (IBDV) were obtained from domestic poultry in New Zealand in 1997 and 1998. An\\u000a in-vivo pathogenicity study carried out in specific pathogen free (SPF) chickens demonstrated the low virulence of one of\\u000a the virus isolates. The nucleotide sequences of the hypervariable region of the VP2 gene of two isolates were determined and\\u000a compared with

Y. F. Chai; N. H. Christensen; C. R. Wilks; J. Meers

2001-01-01

28

High sequence conservation among cucumber mosaic virus isolates from lily.  

PubMed

For classification of Cucumber mosaic virus (CMV) isolates from ornamental crops of different geographical areas, these were characterized by comparing the nucleotide sequences of RNAs 4 and the encoded coat proteins. Within the ornamental-infecting CMV viruses both subgroups were represented. CMV isolates of Alstroemeria and crocus were classified as subgroup II isolates, whereas 8 other isolates, from lily, gladiolus, amaranthus, larkspur, and lisianthus, were identified as subgroup I members. In general, nucleotide sequence comparisons correlated well with geographic distribution, with one notable exception: the analyzed nucleotide sequences of 5 lily isolates showed remarkably high homology despite different origins. PMID:11676424

Chen, Y K; Derks, A F; Langeveld, S; Goldbach, R; Prins, M

2001-08-01

29

Interaction between isolates of barley yellow dwarf virus  

Microsoft Academic Search

1. Some isolates of barley yellow dwarf virus (BYDV) differing in vector transmission characteristics and in host plant reactions were studied in single and mixed inoculations in glass-house trials.2. The symptoms obtained depended on the isolate of BYDV, and the interval between the protective and test inoculation and the variety of host plant.3. Two of the isolates showed complete protection

Harvey C. Smith

1963-01-01

30

Evidence for two groups of banana bunchy top virus isolates  

Microsoft Academic Search

Banana bunchy top virus (BBTV) DNA component 1 from isolates from 10 different countries was cloned and sequenced and the sequences were aligned and com- pared. This analysis indicated two groups: the South Pacific group (isolates from Australia, Burundi, Egypt, Fiji, India, Tonga and Western Samoa) and the Asian group (isolates from the Philippines, Taiwan and Vietnam). The mean sequence

Mirko Karan; Robert M. Harding; James L. Dale

1994-01-01

31

[Isolation of the virus of Syr-Darya Valley fever].  

PubMed

In the course of studies on the ecological structure of acute febrile diseases in the season of activity of blood-sucking arthropods strains of a virus antigenically related to Sikhote-Alyń virus were isolated from the blood of a patient and from Ixodid ticks. This paper presents the results of the study on the causative agent and the clinical picture of the disease caused by this virus. The virus was found to be a new one for science; its appurtenance to the family Picornaviridae, genus Cardiovirus, the antigenic group of encephalomyocarditis has been determined. The virus has been designated "Syr-Darya Valley fever virus" by the area of its isolation. PMID:6097042

L'vov, D K; Karimov, S K; Kiriushchenko, T V; Chun-Siun, F; Skvortsova, T M

1984-01-01

32

Chikungunya virus was isolated in Thailand, 2010.  

PubMed

Chikungunya fever (CHIKF) is an acute febrile illness caused by a mosquito-borne alphavirus, chikungunya virus (CHIKV). This disease re-emerged in Kenya in 2004, and spread to the countries in and around the Indian Ocean. The re-emerging epidemics rapidly spread to regions like India and Southeast Asia, and it was subsequently identified in Europe in 2007, probably as a result of importation of chikungunya cases. On the one hand, chikungunya is one of the neglected diseases and has only attracted strong attention during large outbreaks. In 2008-2009, there was a major outbreak of chikungunya fever in Thailand, resulting in the highest number of infections in any country in the region. However, no update of CHIKV circulating in Thailand has been published since 2009. In this study, we examined the viral growth kinetics and sequences of the structural genes derived from CHIKV clinical isolates obtained from the serum specimens of CHIKF-suspected patients in Central Thailand in 2010. We identified the CHIKV harboring two mutations E1-A226V and E2-I211T, indicating that the East, Central, and South African lineage of CHIKV was continuously circulating as an indigenous population in Thailand. PMID:25113745

Sasayama, Mikiko; Benjathummarak, Surachet; Kawashita, Norihito; Rukmanee, Prasert; Sangmukdanun, Suntaree; Masrinoul, Promsin; Pitaksajjakul, Pannamthip; Puiprom, Orapim; Wuthisen, Pitak; Kurosu, Takeshi; Chaichana, Panjaporn; Maneekan, Pannamas; Ikuta, Kazuyoshi; Ramasoota, Pongrama; Okabayashi, Tamaki; Singhasivanon, Pratap; Luplertlop, Natthanej

2014-12-01

33

Isolation and Phylogenetic Grouping of Equine Encephalosis Virus in Israel  

PubMed Central

During 2008–2009 in Israel, equine encephalosis virus (EEV) caused febrile outbreaks in horses. Phylogenetic analysis of segment 10 of the virus strains showed that they form a new cluster; analysis of segment 2 showed ?92% sequence identity to EEV-3, the reference isolate. Thus, the source of this emerging EEV remains uncertain. PMID:22000361

Aharonson-Raz, Karin; Steinman, Amir; Bumbarov, Velizar; Maan, Sushila; Maan, Narender Singh; Nomikou, Kyriaki; Batten, Carrie; Potgieter, Christiaan; Gottlieb, Yuval; Mertens, Peter

2011-01-01

34

Isolation of 16L Virus: A Rapidly Transforming Sarcoma Virus from an Avian Leukosis Virus-Induced Sarcoma  

Microsoft Academic Search

We have isolated a replication-defective rapidly transforming sarcoma virus (designated 16L virus) from a fibro-sarcoma in a chicken infected with td107A, a transformation-defective deletion mutant of subgroup A Schmidt-Ruppin Rous sarcoma virus. 16L virus transforms fibroblasts and causes sarcomas in infected chickens within 2 wk. Its genomic RNA is 6.0 kilobases and contains sequences homologous to the transforming gene (fps)

Benjamin G. Neel; Lu-Hai Wang; Bernard Mathey-Prevot; Teruko Hanafusa; Hidesaburo Hanafusa; William S. Hayward

1982-01-01

35

Complete genome sequence of a raccoon rabies virus isolate  

Microsoft Academic Search

The entire genome of a mid-Atlantic raccoon strain rabies virus (RRV) isolated in Canada was sequenced; this is the second North American wildlife rabies virus isolate to be fully characterized. The overall organization and length of the genome was similar to that of other lyssaviruses. The nucleotide sequence identity of the raccoon strain ranged between 32.7% and 85.0% when compared

Annamaria G. Szanto; Susan A. Nadin-Davis; Bradley N. White

2008-01-01

36

Identification and evaluation of an isolate of sugarcane mosaic virus  

E-print Network

IDENTIFICATION AND EVALUATION OF AN ISOLATE OF SUGARCANE MOSAIC VIRUS A Thesis by LAURA MARIA GIORDA DE MESSINA Submitted to the Graduate College of Texas ARM University in partial fulfillment of the requirement for the degree of MASt...ER OF SCIENCE May 1983 Major Subject: Plant Pathology IDENTIFICATION AND EVALUATION OF AN ISOLATE OF SUGARCANE MOSAIC VIRUS A Thesis by LAURA MARIA GIORDA DE MESSINA Approved as to style and content by: R. W. loler airman of Committee) ~F. ' R...

Giorda de Messina, Laura Maria

2012-06-07

37

Isolation and characterization of cidofovir resistant vaccinia viruses  

PubMed Central

Background The emergence of drug resistant viruses, together with the possibility of increased virulence, is an important concern in the development of new antiviral compounds. Cidofovir (CDV) is a phosphonate nucleotide that is approved for use against cytomegalovirus retinitis and for the emergency treatment of smallpox or complications following vaccination. One mode of action for CDV has been demonstrated to be the inhibition of the viral DNA polymerase. Results We have isolated several CDV resistant (CDVR) vaccinia viruses through a one step process, two of which have unique single mutations within the DNA polymerase. An additional resistant virus isolate provides evidence of a second site mutation within the genome involved in CDV resistance. The CDVR viruses were 3–7 fold more resistant to the drug than the parental viruses. The virulence of the CDVR viruses was tested in mice inoculated intranasally and all were found to be attenuated. Conclusion Resistance to CDV in vaccinia virus can be conferred individually by at least two different mutations within the DNA polymerase gene. Additional genes may be involved. This one step approach for isolating resistant viruses without serial passage and in the presence of low doses of drug minimizes unintended secondary mutations and is applicable to other potential antiviral agents. PMID:18479513

Becker, Marie N; Obraztsova, Maria; Kern, Earl R; Quenelle, Debra C; Keith, Kathy A; Prichard, Mark N; Luo, Ming; Moyer, Richard W

2008-01-01

38

Genetic Diversity of Argentine Isolates of Feline Immunodeficiency Virus  

Microsoft Academic Search

We report the nucleotide sequence and genetic diversity of part of the envelope (env) gene of four strains of feline immunodeficiency virus (FIV) isol- ated from Argentine domestic cats. The DNA enco- ding the V3 to V5 regions of the env gene of the FIV isolates were amplified by PCR, cloned and sequ- enced. Phylogenetic analysis revealed that the Argentine

Marcelo R. Pecoraro; K. Tomonaga; T. Miyazawa; Y. Kawaguchi; S. Sugita; Y. Tohya; C. Kai; M. E. Etcheverrigaray; T. Mikami

1996-01-01

39

Molecular characterization and heterogeneity of feline immunodeficiency virus isolates  

Microsoft Academic Search

Summary We have molecularly cloned the complete genomic DNA of TM2 strain of feline immunodeficiency virus (FIV) isolated in Japan and compared its nucleotide and the deduced amino acid sequence with those of previously described U.S. isolates, FIV Petaluma and FIV PPR. The infectious molecular clone of FIV TM2 is different from FIV Petaluma in host cell range; the clone

N. Maki; T. Miyazawa; M. Fukasawa; A. Hasegawa; M. Hayami; K. Miki; T. Mikami

1992-01-01

40

Herpes simplex virus: isolation, cytopathological characterization and antiviral sensitivity*  

PubMed Central

BACKGROUND Herpes simplex virus (HSV) infection is an endemic disease and it is estimated that 6095% of the adult population are infected with symptoms that are usually self-limiting, though they can be serious, extensive and prolonged in immunocompromised individuals, highlighted by the emergence of drug-resistant strains. The study of the wild-type HSV strains based on the cytopathogenic features and its antiviral sensitivity are important in the establishment of an antivirogram for controlling the infection. OBJECTIVE This study sought to isolate and examine the cytopathological characteristics of circulating strains of the Herpes simplex virus, from clinical specimens and their sensitivity to commercially available antiherpesvirus drugs, acyclovir, phosphonophormic acid and trifluridine. METHODS Herpes simplex virus isolation, cytopathological features and antiviral sensitivity assays were performed in cell culture by tissue culture infectious dose or plaque forming unit assay. RESULTS From twenty-two clinical specimens, we isolated and adapted nine strains. Overall, the cytopathic effect was detected 24 h post-infection (p.i.) and the presence of syncytia was remarkable 48 h p.i., observed after cell staining. Out of eight isolates, four developed plaques of varying sizes. All the isolates were sensitive to acyclovir, phosphonophormic and trifluridine, with the percentage of virus inhibition (%VI) ranging from 49.7-100%. CONCLUSIONS The methodology for HSV isolation and characterization is a straightforward approach, but the drug sensitivity test, regarded as being of great practical importance, needs to be better understood. PMID:24937819

Nozawa, Carlos; Hattori, Lilian Yumi; Galhardi, Ligia Carla Faccin; Lopes, Nayara; Bomfim, Wesley Andrade; de Cândido, Ligyana Korki; de Azevedo, Elbens Marcos Minoreli; Gon, Airton dos Santos; Linhares, Rosa Elisa Carvalho

2014-01-01

41

First Human Isolate of Hantavirus (Andes virus) in the Americas  

Microsoft Academic Search

We isolated Andes virus (formal name: Andes virus (ANDV), a species in the genus Hantavirus), from serum of an asymptomatic 10-year-old Chilean boy who died 6 days later of hantavirus pulmonary syn- drome (HPS). The serum was obtained 12 days after his grandmother died from HPS and 2 days before he became febrile. No hantavirus immunoglobulin (Ig) G or IgM

Hector Galeno; Judith Mora; Eliecer Villagra; Jorge Fernandez; Jury Hernandez; Gregory J. Mertz; Eugenio Ramirez

2002-01-01

42

Characterization of two strains of tribe? virus isolated in ukraine.  

PubMed

Abstract During arbovirus surveillance in Ixodes ricinus ticks in South Ukraine, two strains of Tribe? virus were isolated using an in vivo method and characterized using the complement fixation text (CFT), hemagglutination assay (HA), electron microscopy, and molecular methods. Both strains replicated well in the BHK-21, PEC, and Vero cell lines and demonstrated 95% nucleotide identity in their VP3 sequences compared with the reference VP3 sequence for Tribe? virus. These two strains of Tribe? virus were named Tr35 and Tr19. Also, segment reassortment involving Tr35, Tr19, TRBV, and LIPV strains was shown. PMID:25409272

Dedkov, Vladimir G; Dubina, Dmitriy A; Yurchenko, Oksana A; Bekova, Marina V; Valdokhina, Anna V; Shipulin, German A

2014-11-01

43

Isolation of the envelope of vesicular stomatitis virus.  

PubMed Central

Vesicular stomatitis virus was disrupted by a combination of freezing and thawing, osmotic shock, and sonic treatment. Subviral components were separated by isopycnic centrifugation. The low-density, lipid-rich fractions were pooled and shown to contain primarily viral glycoprotein. Further purification of this material resulted in the isolation of a preparation of vesicles which contained only the G protein and the same phospholipids as in the intact virions and exhibited spikelike structures similar to those on intact vesicular stomatitis virions. We conclude that we have isolated fragments of native vesicular stomatitis virus envelopes. Images PMID:209217

Taube, S E; Rothfield, L I

1978-01-01

44

Genetic typing of bovine viral diarrhea virus isolates from Argentina  

Microsoft Academic Search

Genetic typing of 29 Bovine Viral Diarrhea Virus (BVDV) isolates from Argentina was carried out by sequencing 245 nucleotides of the RT-PCR products of the 5?-UTR region. Sequence analysis shows that these Argentinean BVDV include types 1 and 2. The majority (26\\/29) of the isolates are type 1, which comprises subtypes 1a and 1b, together with an additional subgroup within

Leandro R Jones; Rubén Zandomeni; E. Laura Weber

2001-01-01

45

Sequence variation and biological activity of rubella virus isolates  

Microsoft Academic Search

Summary Haemagglutination (HA) by rubella virus is mediated by the E1 glycoprotein. Rubella isolates which haemagglutinate with different avidity have been characterised. A significant reduction of HA titre at pH6.0 was observed in one isolate in which isoleucine is substituted for threonine at rubella E1 residue 280. This residue is located in an epitope (EP1) which we have previously identified

P. Londesborough; L. Ho-Terry; G. Terry

1995-01-01

46

Paramyxovirus type 1 infections of racing pigeons: 1 characterisation of isolated viruses  

Microsoft Academic Search

Viruses isolated from field outbreaks of disease in racing pigeons in continental Europe and Great Britain were shown to be identical by serological tests using conventional chicken antisera and mouse monoclonal antibodies. The pigeon viruses showed high levels of cross-reaction to Newcastle disease virus (NDV) in haemagglutination inhibition tests and Madin-Darby bovine kidney cells infected with pigeon virus isolates bound

DJ Alexander; PH Russell

1984-01-01

47

[Characteristics of isolating influenza virus hemagglutinin using purified bromelin].  

PubMed

Gel filtration and ion-exchange chromatography were used to isolate a fraction with the highest proteolytic activity and virtually lacking glycosidases from a commercial bromeline preparation ("Serva", grade B) showing, in addition to proteolytic, also glycosidase activity. The utilization of purified bromeline allows isolation of hemagglutinin without residual neuraminidase activity from influenza virus of various serotypes (H1, H1, and H3). The results are discussed on the basis of an assumption of analogous localization but different conformation accessibility of bromeline-sensitive parts of the light chains of influenza viruses of different subtypes, and a possible role of the carbohydrate component of surface glycoproteins, hemagglutinin and neuraminidase of influenza virus virions. PMID:7293158

Mukazhanova, G N; Siniakov, M S; Kharitonenkov, I G

1981-01-01

48

Molecular Epidemiology of Rabies Virus Isolates in India  

Microsoft Academic Search

In India, rabies is enzootic and is a serious public health and economic problem. India has a large population of stray dogs which, together with a lack of effective control strategies, might have led to the persistence of rabies virus (RV) in the canine population. Our objective was to study the molecular epidemi- ology of RV isolates in India based

T. Nagarajan; B. Mohanasubramanian; E. V. Seshagiri; S. B. Nagendrakumar; M. R. Saseendranath; M. L. Satyanarayana; D. Thiagarajan; P. N. Rangarajan; V. A. Srinivasan

2006-01-01

49

ISOLEMENT D'UN VIRUS HERPS DANS UN LEVAGE DE PIGEONS DE CHAIR (1)  

E-print Network

ISOLEMENT D'UN VIRUS HERPĂ?S DANS UN Ă?LEVAGE DE PIGEONS DE CHAIR (1) H. VINDEVOGEL P. P. PASTORET, G Champs, 1 200 Bruxelles SUMMARY HERPES VIRUS ISOLATED FROM PIGEONS A virus has been isolated from pigeons is pathogenic for pigeons. Lesions produced by intralaryngeal inoculation consists of small necrotic foci

Boyer, Edmond

50

Replication of Pea Enation Mosaic Virus RNA in Isolated Pea Nuclei  

Microsoft Academic Search

Isolated nuclei from healthy pea plants were primed with pea enation mosaic virus (PEMV), southern bean mosaic virus (SBMV), radish mosaic virus (RdMV), tobacco mosaic virus (TMV), PEMV RNA, SBMV RNA, RdMV RNA, or TMV RNA. RNA replication occurred only with PEMV RNA and not with intact PEMV or any of the other viruses or RNAs, as judged by ensuing

C. A. Powell; G. A. de Zoeten

1977-01-01

51

Iguana Virus, a Herpes-Like Virus Isolated from Cultured Cells of a Lizard, Iguana iguana  

PubMed Central

An agent cytopathic for Terrapene and Iguana cell cultures was isolated from spontaneously degenerating cell cultures prepared from a green iguana (Iguana iguana). The agent, designated iguana virus, caused a cytopathic effect (CPE) of a giant cell type, with eosinophilic inclusions commonly observed within giant cell nuclei. Incubation temperature had a marked effect on CPE and on virus release from infected cells. Within the range of 23 to 36 C, low temperatures favored CPE characterized by cytolysis and small giant cell formation, and significant virus release was observed. At warmer temperatures, a purely syncytial type of CPE and total absence of released virus were noted. A unique type of hexagonal eosinophilic cytoplasmic inclusion was observed within syncytia of infected Terrapene cell cultures incubated at 36 C. In vivo studies revealed no evidence of pathogenicity of iguana virus for suckling mice, embryonated hen's eggs, or several species of reptiles and amphibians. Inoculation of iguana virus into young iguanas consistently caused infection that was “unmasked” only when cell cultures were prepared directly from the infected animal. Filtration studies revealed a virion size of >100 nm and <220 nm. Iguana virus is ether-sensitive and, as presumptively indicated by studies of inhibition by bromodeoxyuridine, possesses a deoxyribonucleic type of nucleic acid. The virus characteristics described, as well as electron microscopy observations described in a separate report, indicate that iguana virus is a member of the herpesvirus group. Images PMID:4344303

Clark, H. Fred; Karzon, David T.

1972-01-01

52

Isolation of Newcastle disease virus from a non-avian host (sheep) and its implications.  

PubMed

Newcastle disease virus (NDV) is an avian virus that has not been isolated from naturally infected non-avian and non-human hosts except for one report in which it was isolated from cattle in 1952. We report here for the first time the isolation and identification of NDV from sheep and suggest that this virus be included in the screening of viruses from non-avian hosts. PMID:22543636

Sharma, Bhaskar; Pokhriyal, Mayank; Rai, Gaurava K; Saxena, Meeta; Ratta, Barkha; Chaurasia, Mona; Yadav, Brijesh S; Sen, Arnab; Mondal, Bimelendu

2012-08-01

53

Isolation and characterization of Midway virus: a new tick-borne virus related to Nyamanini.  

PubMed

Midway virus, a new tick-borne virus isolated from two species of Ornithodoros (Alectorobius) ticks of the capensis group (O capensis, O denmarki), is described from Midway, Kure, and Manana islands in the Central Pacific (Hawaiian Archipelago) and from northern Honshu (Japan). Midway virion is enveloped, unusually large, acid and temperature sensitive, and its type of nucleic acid is RNA. Complement-fixation (CF) tests show a close relation of Midway to Nyamanini virus, which has been isolated from ardeid birds and Argas ticks in Africa, the Indian subcontinent, and Southeastern Asia. However, cross-box tests (CF, mouse and tissue culture neutralization, immunofluorescence) show that these two viruses are quite distinct. Midway virus is lethal for newborn Swiss mice inoculated by intracerebral, but not intraperitoneal route. It fails to kill four-week-old mice by either route. Midway virus causes cytopathic effects in BHK-21 cells and titerable plaques in Vero cells. Antibodies to it were prevalent among nestlings of Larus crassirostris (Black-tailed Gull) on Aomatsushima I., but were scarce among those of Nycticorax nycticorax (Black-crowned Night Heron) of the same island. PMID:7153772

Takahashi, M; Yunker, C E; Clifford, C M; Nakano, W; Fujino, N; Tanifuji, K; Thomas, L A

1982-01-01

54

[Rabies virus isolation in the salivary glands of insectivorous bats].  

PubMed

This study determined the presence of the rabies virus in salivary glands, as well as its titre and antigenic characterisation and the level of exposure to the virus from contact between domestic animals and humans. Twenty-six positive brain samples were selected, 80% of which were from the Brazilian free-tailed bat, Tadarida brasiliensis, corresponding to the period 1999-2005. Antigenic characterisation was conducted on a panel of 19 monoclonal antibodies targeting the rabies virus nucleoprotein supplied by the Centers for Disease Control and Prevention in Atlanta in the United States of America. The results revealed a high percentage of isolations in salivary glands (76.9%). Their average titres were compared in a batch of positive samples of brain and salivary glands, giving values of 4.75 and 3.81 respectively (expressed as log LD50/0.03 ml). The isolated viruses corresponded principally to variant 4 associated with T brasiliensis and variant 6 associated with the hoary bat, Lasiurus cinereus, and the red bat, L. borealis, and their respective subvariants. The level of exposure in domestic animals and humans was 50% during the period under study. PMID:20462155

Gury Dohmen, F; Beltrán, F

2009-12-01

55

Xiburema Virus, a Hitherto Undescribed Virus within the Family Rhabdoviridae Isolated in the Brazilian Amazon Region  

PubMed Central

We report here the first complete open reading frame (ORF) genome sequence of Xiburema virus (XIBV), that of strain BE AR362159, isolated from mosquitoes (Sabethes intermedius) in Sena Madureira, Acre state, northern Brazil. All genes showed similarities with those belonging to members of the family Rhabdoviridae. PMID:24948755

Martins, Livia C.; Diniz Junior, Jose Antonio P.; de Almeida Medeiros, Daniele Barbosa; Cardoso, Jedson F.; da Silva, Daisy E. A.; de Oliveira, Layanna F.; de Vasconcelos, Janaina M.; Vianez Junior, Joao Lidio da S. G.; Vasconcelos, Pedro F. C.

2014-01-01

56

Dispatches Isolation and Phylogenetic Characterization of Ebola Viruses Causing Different Outbreaks in Gabon  

E-print Network

Three outbreaks of Ebola hemorrhagic fever have recently occurred in Gabon. Virus has been isolated from clinical materials from all three outbreaks, and nucleotide sequence analysis of the glycoprotein gene of the isolates and virus present in clinical samples has been carried out. These data indicate that each of the three outbreaks should be considered an independent emergence of a different Ebola virus of the Zaire subtype. As in earlier Ebola virus outbreaks, no genetic variability was detected between virus samples taken during an individual outbreak. Since the first recognized outbreaks of human Ebola virus hemorrhagic fever in Africa in the 1970s, biological and genetic differences have been described among the four distinct Ebola viruses isolated: Zaire, Sudan, and Côte d’Ivoire, all isolated from humans, and Reston virus isolated from macaque monkeys from the Philippines (1-5). Serologic evidence has suggested the presence

unknown authors

57

Complete genome sequence of a rabies virus isolated from a human in central african republic.  

PubMed

To validate the feasibility of using next-generation sequencing in an African context, the complete genome of a rabies virus isolated from a human patient was obtained by high-throughput sequencing after virus isolation in mice and random unbiased amplification. Phylogenetic analysis suggested that this virus belongs to the Africa II clade. PMID:24948767

Tricou, Vianney; Berthet, Nicolas; Nakouné, Emmanuel; Kazanji, Mirdad

2014-01-01

58

Complete Genome Sequence of a Rat Hepatitis E Virus Strain Isolated in the United States  

PubMed Central

Hepatitis E virus is a common cause of acute hepatitis in humans. Related viruses have been isolated from multiple animal species, including rats, but their impact on human health is unclear. We present the first full-length genome sequence of a rat hepatitis E virus strain isolated in the United States (LA-B350). PMID:25377700

Debing, Yannick; Emerson, Suzanne U.; Purcell, Robert H.; Dallmeier, Kai

2014-01-01

59

Complete Genome Sequence of a Rabies Virus Isolated from a Human in Central African Republic  

PubMed Central

To validate the feasibility of using next-generation sequencing in an African context, the complete genome of a rabies virus isolated from a human patient was obtained by high-throughput sequencing after virus isolation in mice and random unbiased amplification. Phylogenetic analysis suggested that this virus belongs to the Africa II clade. PMID:24948767

Berthet, Nicolas; Nakoune, Emmanuel; Kazanji, Mirdad

2014-01-01

60

Antigenic and genetic characterization of rabies virus isolates from Uruguay.  

PubMed

After 25 years without any reported cases of rabies in Uruguay, the northern region of the country experienced an epizootic of bovine paralytic rabies in October 2007. The outbreak affected bovines and equines, and the main source of infection was the bat Desmodus rotundus, the only hematophagous species in the country. From October 2007 to July 2008, 42 bovine, 3 equine and 120 chiropteran samples were submitted to the National Veterinary Diagnostic Laboratory for rabies testing. A total of 12 samples (7 bovine, 2 equine and 3 from D. rotundus) were positive by the fluorescent antibody test, and viruses were isolated by the mouse inoculation test. The objective of this study was to compare the antigenic and genetic characteristics of these isolates and three isolates from insectivorous bats from other regions. Antigenic typing using a panel of eight monoclonal antibodies identified all 12 viruses as variant 3 (AgV3), a variant associated with D. rotundus. Two isolates from insectivorous bats (Tadarida brasiliensis and Molossus sp.) were characterized as antigenic variant 4 (AgV4) while the third, from Myotis sp., could not be characterized using this panel as its reactivity pattern did not match that of any of the known antigenic variants. Partial N-gene sequences (nt 149-1420) of these isolates were aligned with homologous sequences derived from GenBank by the CLUSTAL/W method and used to build a neighbor-joining distance tree with the Kimura 2-parameter model. All 12 isolates were genetically grouped into the D. rotundus cluster as they shared 100% identity. In the phylogenetic analysis, the three isolates from insectivorous bats segregated into three clusters: one related to T. brasiliensis, one to Myotis sp. and the other to Lasiurus sp., although the isolate associated with the latter came from a Molossus sp. specimen. These results indicate that AgV3 was associated with the outbreak of bovine paralytic rabies in Uruguay. This is the first report of rabies virus having been detected in non-hematophagous bats in this country. PMID:23318595

Guarino, Helena; Castilho, Juliana Galera; Souto, Juanita; Oliveira, Rafael de Novaes; Carrieri, Maria Luiza; Kotait, Ivanete

2013-05-01

61

Cedar Virus: A Novel Henipavirus Isolated from Australian Bats  

PubMed Central

The genus Henipavirus in the family Paramyxoviridae contains two viruses, Hendra virus (HeV) and Nipah virus (NiV) for which pteropid bats act as the main natural reservoir. Each virus also causes serious and commonly lethal infection of people as well as various species of domestic animals, however little is known about the associated mechanisms of pathogenesis. Here, we report the isolation and characterization of a new paramyxovirus from pteropid bats, Cedar virus (CedPV), which shares significant features with the known henipaviruses. The genome size (18,162 nt) and organization of CedPV is very similar to that of HeV and NiV; its nucleocapsid protein displays antigenic cross-reactivity with henipaviruses; and it uses the same receptor molecule (ephrin- B2) for entry during infection. Preliminary challenge studies with CedPV in ferrets and guinea pigs, both susceptible to infection and disease with known henipaviruses, confirmed virus replication and production of neutralizing antibodies although clinical disease was not observed. In this context, it is interesting to note that the major genetic difference between CedPV and HeV or NiV lies within the coding strategy of the P gene, which is known to play an important role in evading the host innate immune system. Unlike HeV, NiV, and almost all known paramyxoviruses, the CedPV P gene lacks both RNA editing and also the coding capacity for the highly conserved V protein. Preliminary study indicated that CedPV infection of human cells induces a more robust IFN-? response than HeV. PMID:22879820

Barr, Jennifer A.; Tachedjian, Mary; Smith, Craig; Middleton, Deborah; Yu, Meng; Todd, Shawn; Foord, Adam J.; Haring, Volker; Payne, Jean; Robinson, Rachel; Broz, Ivano; Crameri, Gary; Field, Hume E.; Wang, Lin-Fa

2012-01-01

62

Characterization of Whitney's Clethrionomy gapperi virus isolates from Massachusetts.  

PubMed

Six strains of virus were recovered from the blood and/or liver of five Clethrionomys gapperi ochraceus trapped in southeastern Massachusetts during 1969. Biological, antigenic and physiochemical properties of these isolates are reported. USA M-2268a was selected as the reference strain. This strain was identical by complement-fixation and neutralization tests to Whitneys C. gapperie virus (USA 64-7855) from New York State and was related to, but distinct from, an unpublished agent (Johnson's Microtus montanus enterovirus USA M-1146) isolated in June, 1962 from voles trapped in Klamath County, Oregon. USA M-2268a was resistant to lipid solvents and acid pH and was stable at temperatures of 4 C, 22 C, and 37 C. Virus was detected over a 10-day observation period in four species of mosquitoes inoculated with USA M-2268a, although there was no evidence of infection or replication, and transmission attempts by bite failed. Neutralizing antibody was detected in C.g. gapperi and C. g. ochraceus in various habitats throughout the state. PMID:6801

Main, A J; Shope, R E; Wallis, R C

1976-04-01

63

Genomic characterization of pseudorabies virus strains isolated in Italy.  

PubMed

In this study, we undertook the genomic characterization of 54 pseudorabies virus (PRV) strains isolated in Italy during 1984-2010. The characterization was based on partial sequencing of the UL44 (gC) and US8 (gE) genes; 44 strains (38 for gene gE and 36 for gC) were isolated on pig farms; 9 originated from dogs and 1 from cattle. These porcine PRV strains, which were closely related to those isolated in Europe and America in the last 20 years, and the bovine strain bovine/It/2441/1992 belong to cluster B in both phylogenetic trees. Six porcine strains that do not belong to cluster B are related in both gE and gC phylogenetic trees to the 'old' porcine PRV strains isolated in the 1970s and 1980s. In the last two decades, the presence of these strains in domestic pig populations has been reduced drastically, whereas they are prevalent in wild boar. The two remaining strains have an interesting genomic profile, characterized by the gC gene being closely related to the old porcine PRV strains, and the gE gene being similar to that of recently isolated strains. Three strains originating from working dogs on pig farms are located in cluster B in both phylogenetic trees. Five strains isolated from hunting dogs have a high degree of correlation with PRV strains circulating in wild boar. The last isolate has a gC gene similar to that in the two porcine strains mentioned previously, and the gE gene is correlated with the strains isolated from hunting dogs. These results provide interesting insight into the genomic characterization of PRV strains and reveal a clear differentiation between the strains isolated from hunting dogs that are related to the wild boar strains and those originating from domestic pigs. PMID:23331342

Sozzi, E; Moreno, A; Lelli, D; Cinotti, S; Alborali, G L; Nigrelli, A; Luppi, A; Bresaola, M; Catella, A; Cordioli, P

2014-08-01

64

Recovery of H14 influenza A virus isolates from sea ducks in the Western Hemisphere  

PubMed Central

In 2010, H14 influenza A viruses were recovered from clinically normal sea ducks in the United States. These are the first H14 isolates recovered in the Western Hemisphere and represent the only documented H14 influenza A viruses isolated since the original isolates were recovered from near the Caspian Sea during 1982. PMID:22307173

Nolting, Jacqueline; Fries, Anthony C.; Slemons, Richard D; Courtney, Chad; Hines, Nichole; Pedersen, Janice

2012-01-01

65

Endogenous mink (Mustela vison) type C virus isolated from sarcoma virus-transformed mink cells.  

PubMed Central

A previously described type virus stock (designated PP-1R), isolated by cocultivating baboon cells with mink cells transformed by Kirsten sarcoma virus (64J1), has been further cloned and characterized. End point-diluted stocks of PP-1R have been obtained that are free of focus-forming activity and lack both Kirsten sarcoma and primate type C viral sequences. Nucleic acid hybridization experiments show that the cloned virus (MiLV) is an endogenous, genetically transmitted virus of the mink (Mustela vison). MiLV replicates in canine, feline, and 64J1 mink cells but not in an untransformed mink cell line. Multiple viral gene copies can be detected in the DNA of normal mink cells in culture and in normal mink tissues; related endogenous viral genes are also detected in several related Mustela species. The virus codes for a p30 protein very closely related antigenically to that of feline leukemia virus but contains p15 and p12 proteins that are antigenically distinct. The mink cell line, Mv1Lu, and its Kirsten sarcoma-transformed derivatives, 64J1, express relatively low levels of type C viral RNA related to MiLV and normally do not produce detectable levels of MiLV p30 protein or complete, infectious viral particles. Infection of sarcoma virus-transformed mink cells with baboon type C virus, however, can augment the level of expression of endogenous mink viral RNA and can result in the synthesis and packaging of mink viral RNA and p30 antigen in extracellular virions. Since the Mv1Lu cell line and its tranformed derivatives have become widely used in studies of retroviruses, the possibility of activating endogenous mink viral genes should be considered by investigators working with these cells. PMID:76684

Sherr, C J; Benveniste, R E; Todaro, G J

1978-01-01

66

Isolations of Potosi virus from mosquitoes collected in the United States, 1989-94.  

PubMed

Potosi (POT) virus, a recently characterized Bunyamwera serogroup virus, was discovered when it was isolated from Aedes albopictus collected at a waste-tire site in Potosi, Washington County, Missouri, during 1989. During the following year, POT virus was not isolated from 39,048 mosquitoes, including 17,519 Ae. albopictus, collected in Washington County. In 1991, mosquito collections from South Carolina, Ohio, and Michigan yielded 8 strains of POT virus: 6 from Coquillettidia perturbans and one each from Culex restuans and Psorophora columbiae. Additional collections of Ae. albopictus from several states during 1990-93 failed to yield further isolates of POT virus. In 1994, POT virus was isolated from Ae. albopictus and Anopheles punctipennis in North Carolina and from Ae. albopictus in Illinois. These represent the first virus isolations of any type for Ae. albopictus in those states. Thus far, POT virus has been isolated from 5 mosquito species in different genera in 6 states. The known geographic range of POT virus, based on virus isolations, has been extended from Missouri to the upper Midwest and the Atlantic seaboard. Potential vector relationships and possible transmission cycles of POT virus are discussed. PMID:8723251

Mitchell, C J; Smith, G C; Karabatsos, N; Moore, C G; Francy, D B; Nasci, R S

1996-03-01

67

First isolation of a giant virus from wild Hirudo medicinalis leech: Mimiviridae isolation in Hirudo medicinalis.  

PubMed

Giant viruses and amoebae are common in freshwater, where they can coexist with other living multicellular organisms. We screened leeches from the species Hirudo medicinalis for giant viruses. We analyzed five H. medicinalis obtained from Tunisia (3) and France (2). The leeches were decontaminated and then dissected to remove internal parts for co-culture with Acanthamoeba polyphaga. The genomes of isolated viruses were sequenced on a 454 Roche instrument, and a comparative genomics analysis was performed. One Mimivirus was isolated and the strain was named Hirudovirus. The genome assembly generated two scaffolds, which were 1,155,382 and 25,660 base pairs in length. Functional annotations were identified for 47% of the genes, which corresponds to 466 proteins. The presence of Mimividae in the same ecological niche as wild Hirudo may explain the presence of the mimivirus in the digestive tract of the leech, and several studies have already shown that viruses can persist in the digestive tracts of leeches fed contaminated blood. As leeches can be used medically and Mimiviruses have the potential to be an infectious agent in humans, patients treated with leeches should be surveyed to investigate a possible connection. PMID:24287596

Boughalmi, Mondher; Pagnier, Isabelle; Aherfi, Sarah; Colson, Philippe; Raoult, Didier; La Scola, Bernard

2013-12-01

68

First Isolation of a Giant Virus from Wild Hirudo medicinalis Leech: Mimiviridae isolation in Hirudo medicinalis  

PubMed Central

Giant viruses and amoebae are common in freshwater, where they can coexist with other living multicellular organisms. We screened leeches from the species Hirudo medicinalis for giant viruses. We analyzed five H. medicinalis obtained from Tunisia (3) and France (2). The leeches were decontaminated and then dissected to remove internal parts for co-culture with Acanthamoeba polyphaga. The genomes of isolated viruses were sequenced on a 454 Roche instrument, and a comparative genomics analysis was performed. One Mimivirus was isolated and the strain was named Hirudovirus. The genome assembly generated two scaffolds, which were 1,155,382 and 25,660 base pairs in length. Functional annotations were identified for 47% of the genes, which corresponds to 466 proteins. The presence of Mimividae in the same ecological niche as wild Hirudo may explain the presence of the mimivirus in the digestive tract of the leech, and several studies have already shown that viruses can persist in the digestive tracts of leeches fed contaminated blood. As leeches can be used medically and Mimiviruses have the potential to be an infectious agent in humans, patients treated with leeches should be surveyed to investigate a possible connection. PMID:24287596

Boughalmi, Mondher; Pagnier, Isabelle; Aherfi, Sarah; Colson, Philippe; Raoult, Didier; La Scola, Bernard

2013-01-01

69

Isolation of H9N2 avian influenza virus from bobwhite quail (Colinus virginianus) in Egypt.  

PubMed

This study describes the first isolation of H9N2 avian influenza virus (AIV) from commercial bobwhite quail (Colinus virginianus) in Egypt. Infected birds showed neither clinical signs nor mortality. Virus isolation and real-time reverse transcription polymerase chain reaction confirmed the presence of the H9N2 virus in cloacal swab samples collected at 35 days of age and the absence of other AIV subtypes, including H5 and H7. The hemagglutinin and neuraminidase genes of the isolated virus showed 99.1% and 98.2% nucleotide identity and 97.3% and 100% amino acid identity, respectively, to those of H9N2 viruses currently circulating in poultry in the Middle East. Phylogenetically, the Egyptian H9N2 virus was closely related to viruses of the G1-like lineage isolated from neighbouring countries, indicating possible epidemiological links. PMID:22426861

El-Zoghby, Elham F; Arafa, Abdel-Satar; Hassan, Mohamed K; Aly, Mona M; Selim, Abdullah; Kilany, Walid H; Selim, Usama; Nasef, Soad; Aggor, Mohamed G; Abdelwhab, E M; Hafez, Hafez M

2012-06-01

70

Isolation and partial characterisation of a new strain of Ebola We have isolated a new strain of Ebola virus from a non-  

E-print Network

1271 Isolation and partial characterisation of a new strain of Ebola virus Summary We have isolated a new strain of Ebola virus from a non- fatal human case infected during the autopsy of a wild about the natural reservoir of the Ebola virus. Lancet 1995; 345: 1271-74 Introduction Ebola virus

71

Clinical, pathological and antigenic aspects of bovine viral diarrhea virus (BVDV) type 2 isolates identified in Brazil  

Microsoft Academic Search

Nucleotide sequencing and phylogenetic analysis of Brazilian bovine viral diarrhea virus (BVDV) field isolates identified four viruses belonging to the genotype 2. Comparison of 5? UTR sequences from these isolates to those of North American BVDV type 2 revealed genomic variations that correlated with the geographic origins of the isolates. Two of the Brazilian type 2 viruses were isolated from

E. F Flores; L. H. G. V Gil; S. A Botton; R Weiblen; J. F Ridpath; L. C Kreutz; C Pilati; D Driemeyer; V Moojen; A. C Wendelstein

2000-01-01

72

First isolation of reticuloendotheliosis virus from mallards in China.  

PubMed

Reticuloendotheliosis virus (REV) causes an oncogenic, immunosuppressive and runting syndrome in many avian hosts worldwide. REV infection has never been reported in mallard ducks, however. To identify REV infection in mallards, we collected 40 mallard duck samples from Jilin Province of China. In this study, the REV strain, DBYR1102, was first isolated from a mallard in China and identified by PCR, indirect immunofluorescence assay and electron microscopy. The gp90 gene and complete LTR of DBYR1102 were amplified and sequenced. Phylogenetic analysis based on gp90 genes of REV indicated that the REV strain DBYR1102 is closely related to strain HLJR0901 from northeastern China, the prairie chicken isolate APC-566, and REV subtype III, represented by chick syncytial virus. This new strain is distantly related to two other subtypes of REV, 170A and SNV. Phylogenetic analysis based on the LTR yielded information similar to that obtained with the gp90 genes. The results of this study not only expand our epidemiological understanding of REV in the wild birds of China but also demonstrate the potential role of wild waterfowl in REV transmission. PMID:24643331

Jiang, Lili; Deng, Xiaoyun; Gao, Yulong; Li, Kai; Chai, Hongliang; Fan, Zhaobin; Ren, Xiangang; Wang, Qi; Zhang, Lizhou; Yun, Bingling; Yin, Chunhong; Chen, Yuming; Qin, Liting; Gao, Honglei; Wang, Yongqiang; Hua, Yuping; Wang, Xiaomei

2014-08-01

73

Isolation of Borna Disease Virus from Human Brain Tissue  

PubMed Central

Serological and molecular epidemiological studies indicate that Borna disease virus (BDV) can infect humans and is possibly associated with certain neuropsychiatric disorders. We examined brain tissue collected at autopsy from four schizophrenic patients and two healthy controls for the presence of BDV markers in 12 different brain regions. BDV RNA and antigen was detected in four brain regions of a BDV-seropositive schizophrenic patient (P2) with a very recent (2 years) onset of disease. BDV markers exhibited a regionally localized distribution. BDV RNA was found in newborn Mongolian gerbils intracranially inoculated with homogenates from BDV-positive brain regions of P2. Human oligodendroglia (OL) cells inoculated with brain homogenates from BDV-positive gerbils allowed propagation and isolation of BDVHuP2br, a human brain-derived BDV. Virus isolation was also possible by transfection of Vero cells with ribonucleoprotein complexes prepared from BDV-positive human and gerbil brain tissues. BDVHuP2br was genetically closely related to but distinct from previously reported human- and animal-derived BDV sequences. PMID:10775596

Nakamura, Yurie; Takahashi, Hirokazu; Shoya, Yuko; Nakaya, Takaaki; Watanabe, Makiko; Tomonaga, Keizo; Iwahashi, Kazuhiko; Ameno, Kiyoshi; Momiyama, Noriko; Taniyama, Hiroyuka; Sata, Tetsutaro; Kurata, Takeshi; de la Torre, Juan Carlos; Ikuta, Kazuyoshi

2000-01-01

74

Serotyping and strain identification of maize streak virus isolates.  

PubMed

Four strains of maize streak virus, namely the Panicum maximum, Digitaria setigera and sugarcane strains, have been identified from 19 isolates by ELISA using polyclonal antisera cross-absorbed with particles of the maize strain. The results suggest there is an epitope of the maize strain which is not dependent on the capsid being intact and which is common to all the members of the group; other strain-specific epitopes are probably conformation-dependent. A specific epitope (probably internal) occurs on the coat protein of a maize strain isolate, D(R)D, grown in D. velutina, that is also present on the coat protein of the D. setigera (previously reported as D. sanguinalis) strain. Specific internal epitopes also occur in the coat proteins of sugarcane and P. maximum strains. The use of indirect ELISA was necessary for accurate serotyping. The serological reactivities of particles of all the members of each type were identical irrespective of the host from which they were extracted. Sap extracts proved to be more reliable sources of antigen than virus preparations, which could vary in their serological reactivity. Nevertheless, serological differentiation index values determined in tests using either type of antigen proved reliable and consistent. PMID:1697327

Pinner, M S; Markham, P G

1990-08-01

75

Genotyping of Korean isolates of infectious hematopoietic necrosis virus (IHNV) based on the glycoprotein gene  

USGS Publications Warehouse

Glycoprotein (G) gene nucleotide sequences of four Korean isolates of infectious hematopoietic necrosis virus (IHNV) were analyzed to evaluate their genetic relatedness to worldwide isolates. All Korean isolates were closely related to Japanese isolates of genogroup JRt rather than to those of North American and European genogroups. It is believed that Korean IHNV has been most likely introduced from Japan to Korea by the movement of contaminated fish eggs. Among the Korean isolates, phylogenetically distinct virus types were obtained from sites north and south of a large mountain range, suggesting the possibility of more than one introduction of virus from Japan. ?? 2007 Springer-Verlag.

Kim, W. -S.; Oh, M. -J.; Nishizawa, T.; Park, J. -W.; Kurath, G.; Yoshimizu, M.

2007-01-01

76

Isolation of a Novel Swine Influenza Virus from Oklahoma in 2011 Which Is Distantly Related to Human Influenza C Viruses  

PubMed Central

Of the Orthomyxoviridae family of viruses, only influenza A viruses are thought to exist as multiple subtypes and has non-human maintenance hosts. In April 2011, nasal swabs were collected for virus isolation from pigs exhibiting influenza-like illness. Subsequent electron microscopic, biochemical, and genetic studies identified an orthomyxovirus with seven RNA segments exhibiting approximately 50% overall amino acid identity to human influenza C virus. Based on its genetic organizational similarities to influenza C viruses this virus has been provisionally designated C/Oklahoma/1334/2011 (C/OK). Phylogenetic analysis of the predicted viral proteins found that the divergence between C/OK and human influenza C viruses was similar to that observed between influenza A and B viruses. No cross reactivity was observed between C/OK and human influenza C viruses using hemagglutination inhibition (HI) assays. Additionally, screening of pig and human serum samples found that 9.5% and 1.3%, respectively, of individuals had measurable HI antibody titers to C/OK virus. C/OK virus was able to infect both ferrets and pigs and transmit to naive animals by direct contact. Cell culture studies showed that C/OK virus displayed a broader cellular tropism than a human influenza C virus. The observed difference in cellular tropism was further supported by structural analysis showing that hemagglutinin esterase (HE) proteins between two viruses have conserved enzymatic but divergent receptor-binding sites. These results suggest that C/OK virus represents a new subtype of influenza C viruses that currently circulates in pigs that has not been recognized previously. The presence of multiple subtypes of co-circulating influenza C viruses raises the possibility of reassortment and antigenic shift as mechanisms of influenza C virus evolution. PMID:23408893

Hause, Ben M.; Ducatez, Mariette; Collin, Emily A.; Ran, Zhiguang; Liu, Runxia; Sheng, Zizhang; Armien, Anibal; Kaplan, Bryan; Chakravarty, Suvobrata; Hoppe, Adam D.; Webby, Richard J.; Simonson, Randy R.; Li, Feng

2013-01-01

77

In vitro and in vivo characterization of herpes simplex virus clinical isolates recovered from patients infected with human immunodeficiency virus.  

PubMed Central

A total of 100 herpes simplex viruses isolated from lesions not responding to acyclovir (ACV) therapy were recovered from 51 patients infected with human immunodeficiency virus. In vitro analysis of these isolates included testing their susceptibility to ACV and determining their thymidine kinase (TK) phenotypes. Of the 100 isolates evaluated, 23 were ACV sensitive and 77 were ACV resistant. Seventy-four of these ACV-resistant isolates were of the TK-deficient or low-TK-producer phenotype and three were of the TK-altered phenotype. The TKs isolates that represented each of the different autoradiographic phenotypes were further characterized by enzyme kinetics. The ability of selected isolates to cause disease in vivo was evaluated by using several mouse virulence models. Cutaneous virulence in normal and immunocompromised mice was evaluated, and neurovirulence in normal mice was determined. Latent infections were assayed by the cocultivation of trigeminal ganglia recovered from mice that had survived acute infection. These reactivated viruses were evaluated in vitro and compared with the original infecting isolate. The mechanisms of resistance and pathogenicity of these herpes simplex virus isolates recovered from patients positive for human immunodeficiency virus are similar to those reported for isolates recovered from normal and immunocompromised patients without AIDS. PMID:1666496

Hill, E L; Hunter, G A; Ellis, M N

1991-01-01

78

New Genome Sequences of Gamboa Viruses (Family Bunyaviridae, Genus Orthobunyavirus) Isolated in Panama and Argentina  

PubMed Central

We describe here the nearly complete open reading frame (ORF) of five Gamboa virus strains isolated in Panama and Argentina. The viruses with complete ORF showed the regular genome organization observed in other orthobunyaviruses with exception to the presence of NSs protein. All predicted proteins showed homology with viruses belonging to members of the family Bunyaviridae. PMID:25414487

de Lima, Clayton P. S.; Martins, Lívia C.; Aragăo Dias, Amarílis; Cardoso, Jedson F.; Silva, Sandro P.; Da Silva, Daisy E. A.; Oliveira, Layanna F.; Vasconcelos, Janaina M.; Ferreira, Joăo Paulo C.; Travassos da Rosa, Amelia P. A.; Guzman, Hilda; Tesh, Robert B.; Vasconcelos, Pedro F. C.

2014-01-01

79

Isolation and characterisation of an H3N8 equine influenza virus in Australia, 2007.  

PubMed

Before 2007, equine influenza had never been diagnosed in Australia. On 22 August 2007, infection was confirmed in horses at Eastern Creek Animal Quarantine Station near Sydney. The virus subsequently isolated (A/equine/Sydney/2888-8/2007) was confirmed by sequence analysis of the haemagglutinin (HA) gene as an H3 virus of the variant American Florida lineage that is now referred to as Clade 1. The HA sequence of the virus was identical to that of a virus isolated from a contemporaneous outbreak in Japan and showed high homology to viruses circulating in North America. PMID:21711282

Watson, J; Halpin, K; Selleck, P; Axell, A; Bruce, K; Hansson, E; Hammond, J; Daniels, P; Jeggo, M

2011-07-01

80

Matrix Gene of Influenza A Viruses Isolated from Wild Aquatic Birds: Ecology and Emergence of Influenza A Viruses  

PubMed Central

Wild aquatic birds are the primary reservoir of influenza A viruses, but little is known about the viruses' gene pool in wild birds. Therefore, we investigated the ecology and emergence of influenza viruses by conducting phylogenetic analysis of 70 matrix (M) genes of influenza viruses isolated from shorebirds and gulls in the Delaware Bay region and from ducks in Alberta, Canada, during >18 years of surveillance. In our analysis, we included 61 published M genes of isolates from various hosts. We showed that M genes of Canadian duck viruses and those of shorebird and gull viruses in the Delaware Bay shared ancestors with the M genes of North American poultry viruses. We found that North American and Eurasian avian-like lineages are divided into sublineages, indicating that multiple branches of virus evolution may be maintained in wild aquatic birds. The presence of non-H13 gull viruses in the gull-like lineage and of H13 gull viruses in other avian lineages suggested that gulls' M genes do not preferentially associate with the H13 subtype or segregate into a distinct lineage. Some North American avian influenza viruses contained M genes closely related to those of Eurasian avian viruses. Therefore, there may be interregional mixing of the two clades. Reassortment of shorebird M and HA genes was evident, but there was no correlation among the HA or NA subtype, M gene sequence, and isolation time. Overall, these results support the hypothesis that influenza viruses in wild waterfowl contain distinguishable lineages of M genes. PMID:15280485

Widjaja, Linda; Krauss, Scott L.; Webby, Richard J.; Xie, Tao; Webster, Robert G.

2004-01-01

81

ISOLATION OF BOVINE VIRAL DIARRHEA VIRUS FROM A FREE RANGING MULE DEER IN WYOMING  

Microsoft Academic Search

A noncytopathic type 1a bovine viral diarrhea virus (BVDV) was isolated from a free- ranging yearling female mule deer (Odocoileus hemionus) from northwestern Wyoming (USA). The mule deer was emaciated, weak, and salivating, and Arcanobacterium pyogenes was cultured from lung abscesses. Bovine viral diarrhea virus was isolated from lung, however, BVDV antigen was not detected by immunohistochemistry. The BVDV genotype

Hana Van Campen; Julia Ridpath; Elizabeth Williams; Jacqueline Cavender; Joan Edwards; Scott Smith; Hall Sawyer

82

Virulence Markers in the 5? Untranslated Region of Genotype 2 Bovine Viral Diarrhea Virus Isolates  

Microsoft Academic Search

Virulence markers to distinguish high from low virulence bovine viral diarrhea virus genotype 2 isolates have not been previously reported. The objective of this study was to identify virulence markers by evaluating the primary and secondary structures of the 5?-untranslated region of low and high virulence bovine viral diarrhea virus genotype 2 isolates. The nucleotide sequences of the entire 5?-untranslated

Christina L Topliff; Clayton L Kelling

1998-01-01

83

Respiratory Syncytial Virus Isolation by Combined Continuous Flow-Isopycnic Banding Centrifugation  

PubMed Central

A new zonal centrifuge rotor (B-IX) which combines continuous sample flow centrifugation with isopycnic banding has been used to isolate and concentrate respiratory syncytial virus from liter volumes of culture fluid. This isolation technique utilizes a sucrose density gradient to trap and isopycnically band the virus particles, and permits recovery of the particles from the rotor in an unaggregated condition. PMID:5621468

Cline, G. B.; Coates, Helen; Anderson, N. G.; Chanock, R. M.; Harris, W. W.

1967-01-01

84

Purification, Identity and Some Properties of An Isolate of Barley Yellow Dwarf Virus from Indiana  

Microsoft Academic Search

SUMMARY Methods were developed for preparing an Indiana isolate of barley yellow dwarf virus in amounts (1 to 2 mg\\/kg) and purity comparable to those recently achieved with other luteoviruses. Choice of host, and environmental conditions, period of infection and tissue used were critical factors for virus production. Aphid transmission, serological tests, and other properties defined the isolate as 'PAV-like'

J. Hammond; R. M. Lister; J. E. Foster

1983-01-01

85

Isolations of Potosi Virus from Mosquitoes (Diptera: Culicidae) Collected in Connecticut  

Microsoft Academic Search

Potosi virus (POTV) (Bunyaviridae: Orthobunyavirus) was Ţrst isolated from Aedes albopictus (Skuse) collected in Potosi, MO, in 1989, and subsequent isolations were reported from Illinois, Michigan, Ohio, and the Carolinas. To determine whether the distribution of this virus extends into the northeastern United States, we analyzed arboviruses acquired from mosquitoes collected in Connecticut from 1998 to 2004. In 2001, a

Philip M. Armstrong; Theodore G. Andreadis; John F. Anderson; Andrew J. Main

2005-01-01

86

Isolation of caprine arthritis encephalitis virus from goats in Mexico.  

PubMed Central

A lentivirus was isolated from 2 goats in Mexico that were seropositive to caprine arthritis encephalitis virus (CAEV) by the agar gel immunodiffusion (AGID) test. The lentivirus was identified as CAEV by the observation of giant multinucleated cells (syncytia) in goat synovial membrane (GSM) monolayers co-cultivated with blood mononuclear (BMN) cells from the seropositive goats, and by amplifying a DNA segment of the CAEV gag gene using the polymerase chain reaction (PCR) technique. Subsequently, cell supernatants from the GSM cells co-cultivated with BMN cells were used to infect 2 CAEV-seronegative goats. These goats seroconverted to CAEV as determined by the AGID test, and CAEV was re-isolated from these goats. One of the goats developed polyarthritis 8 mo after inoculation. Previous serological surveys indicate that infection with CAEV is prevalent among goats in Mexico. To our knowledge this is the first report of CAEV isolation in Mexico. Because of globalization of markets and increased trading among nations, the rapid identification and reporting of diseases such as CAEV are important to prevent the dissemination of these diseases. Images Figure 1. Figure 2. PMID:10480464

Daltabuit Test, M; de la Concha-Bermejillo, A; Espinosa, L E; Loza Rubio, E; Aguilar Setien, A

1999-01-01

87

A new host record: strawberry latent ringspot virus isolated from flowering cherry  

Microsoft Academic Search

Strawberry latent ringspot virus (SLRV) was isolated from flowering cherry (Prunus serrulata sensu lato) trees in close proximity to each other in Auckland, New Zealand. The virus was not isolated from any of 390 flowering cherry\\u000a trees tested from four other regions in the North Island. The virus was identified by host range, particle morphology, RNA\\u000a and protein content and

K. R. Everett-v; K. S. Milne; R. L. S. Forster

1994-01-01

88

Leakey Virus: a New Hantavirus Isolated from Mus musculus in the United States  

Microsoft Academic Search

SUMMARY A hantavirus, designated Leakey virus, was isolated from a Mus musculus captured in Real County, Texas, U.S.A. in August 1986. Virus-specific fluorescence was first detected 13 days after inoculation of Vero-E6 cells with spleen tissue from the seropositive M. musculus. Ultrastructurally, the new isolate resembled other hantaviruses. Leakey virus induced a fatal meningoencephalitis in infant Fischer rats, with viral

L. J. Baek; R. Yanagihara; C. J. Gibbs; M. Miyazaki; D. C. Gajdusek

1988-01-01

89

Chemokine Coreceptor Usage by Diverse Primary Isolates of Human Immunodeficiency Virus Type 1  

Microsoft Academic Search

We tested chemokine receptor subset usage by diverse, well-characterized primary viruses isolated from peripheral blood by monitoring viral replication with CCR1, CCR2b, CCR3, CCR5, and CXCR4 U87MG.CD4 transformed cell lines and STRL33\\/BONZO\\/TYMSTR and GPR15\\/BOB HOS.CD4 transformed cell lines. Primary viruses were isolated from 79 men with confirmed human immunodeficiency virus type 1 (HIV-1) infection from the Chicago component of the

LINQI ZHANG; TIAN HE; YAOXING HUANG; ZHIWEI CHEN; YOUNG GUO; SAM WU; KEVIN J. KUNSTMAN; R. CLARK BROWN; JOHN P. PHAIR; AVIDAN U. NEUMANN; DAVID D. HO; STEVEN M. WOLINSKY

1998-01-01

90

[Khurdun virus, a presumably new RNA-containing virus associated with coots (Fulica atra), isolated in the Volga river delta].  

PubMed

The prototype strain LEIV-Ast 01-5 of the unclassified enveloped RNA-containing Khurdun virus, less than 220 nm in size, which is widely distributed among coots (Fulica atra) in the Volga River delta, was deposited on November 4, 2004, at the State Virus Collection (SVC # 992). The virus was isolated annually (2001-2004) with a frequency of 2.3-8.5% (mean 6.3%) when examining 348 coots caught in the lower and middle zones of the Volga River delta. Virological examinations used mixed pools of the brain and spleen to inoculate neonatal albino mice and the cellular line Vero-E6. One strain was isolated from a pygmy cormorant (Phalacrocorax pygmaeus). The virus could not be isolated from other species of 1080 birds, 20 hares, 140,000 mosquitoes of 5 predominant species, and 6,700 Hyalomma marginatum ticks. Any antigenic relationship of the virus with all the viruses early isolated in the Northern Caspian Sea region has not been found. ELISA has been developed to detect and identify Khurdun virus antigen. PMID:16104519

Galkina, I V; L'vov, L N; Gromashevski?, V L; Moskvina, T M

2005-01-01

91

Cucumber mosaic virus groups IA and II are represented among isolates from naturally infected lilies  

Microsoft Academic Search

Four cucumber mosaic virus isolates (named Cas, CB, P26 and Simp2) found in naturally infected lily plants were characterized\\u000a on the basis of their serological properties and the results of analysis of RNA3 sequence fragments containing coat protein\\u000a and movement protein genes. The properties of lily isolates were compared with those of eight other virus isolates originating\\u000a from dahlia, delphinium,

Hanna Berniak; Maria Kami?ska; Tadeusz Malinowski

2010-01-01

92

Isolation of a new herpes virus from human CD4 sup + T cells  

SciTech Connect

A new human herpes virus has been isolated from CD4{sup +} T cells purified from peripheral blood mononuclear cells of a healthy individual (RK), following incubation of the cells under conditions promoting T-cell activation. The virus could not be recovered from nonactivated cells. Cultures of lymphocytes infected with the RK virus exhibited a cytopathic effect, and electron microscopic analyses revealed a characteristic herpes virus structure. RK virus DNA did not hybridize with large probes derived from herpes simplex virus, Epstein-Barr virus, varicella-zoster virus, and human cytomegalovirus. The genetic relatedness of the RK virus to the recently identified T-lymphotropic human herpes virus 6 (HHV-6) was investigated by restriction enzyme analyses using 21 different enzymes and by blot hydridization analyses using 11 probes derived from two strains of HHV-6 (Z29 and U1102). Whereas the two HHV-6 strains exhibited only limited restriction enzyme polymorphism, cleavage of the RK virus DNA yielded distinct patterns. Of the 11 HHV-6 DNA probes tested, only 6 cross-hybridized with DNA fragments derived from the RK virus. Taken together, the maximal homology amounted to 31 kilobases of the 75 kilobases tested. The authors conclude that the RK virus is distinct from previously characterized human herpesviruses. The authors propose to designate it as the prototype of a new herpes virus, the seventh human herpes virus identified to date.

Frenkel, N.; Schirmer, E.C.; Wyatt, L.S.; Katsafanas, G.; Roffman, E.; Danovich, R.M. (National Institutes of Health, Rockville, MD (USA)); June, C.H. (Naval Medical Research Institute, Bethesda, MD (USA))

1990-01-01

93

Cedar Virus: A Novel Henipavirus Isolated from Australian Bats  

Microsoft Academic Search

The genus Henipavirus in the family Paramyxoviridae contains two viruses, Hendra virus (HeV) and Nipah virus (NiV) for which pteropid bats act as the main natural reservoir. Each virus also causes serious and commonly lethal infection of people as well as various species of domestic animals, however little is known about the associated mechanisms of pathogenesis. Here, we report the

Glenn A. Marsh; Carol de Jong; Jennifer A. Barr; Mary Tachedjian; Craig Smith; Deborah Middleton; Meng Yu; Shawn Todd; Adam J. Foord; Volker Haring; Jean Payne; Rachel Robinson; Ivano Broz; Gary Crameri; Hume E. Field; Lin-Fa Wang

2012-01-01

94

Isolation and propagation of infectious bursal disease virus using the ovine kidney continuous cell line.  

PubMed

Twenty-six samples known to contain infectious bursal disease virus (IBDV) were examined by virus-isolation attempts on ovine kidney (OK) cell line, Vero cell line, and chicken embryo fibroblast (CEF) cultures. Virus was isolated from two of 26 samples, three of 26 samples, and three of 25 samples on OK, Vero, and CEF cultures, respectively. However, in contrast to IBDV replication in Vero and CEF cultures, isolated virus was unable to induce serially sustained cytopathic effects (CPE) during successive passages in the OK cell line, unless cell lysates were treated with chloroform between every other passage. The cytopathogenicity of the untreated virus passaged in OK cells was revived and maintained upon passage in Vero cells. An initial single passage of laboratory or field material in OK cells followed by further passages in Vero cells resulted in virus isolation from six of 26 samples, which was better virus recovery than when either cell line was used alone or when CEF cultures were used. Twenty of the 26 test samples were originally positive when examined by nucleic acid hybridization with radiolabeled IBDV cDNA, indicating that some of the samples that were negative upon virus isolation using OK and Vero cells may have contained inactivated virus. PMID:1320862

Kibenge, F S; McKenna, P K

1992-01-01

95

Rabies virus isolates of India - Simultaneous existence of two distinct evolutionary lineages.  

PubMed

Rabies is a fatal viral disease of serious public health implication. The disease is enzootic in India. In the present study, thirty six rabies virus isolates were obtained from terrestrial mammals of India during 2002-2012. Ecto-domain coding region of the glycoprotein gene from all the isolates were sequenced and the phylogenetic analysis was performed in relation to the global rabies and rabies related virus isolates. The Indian isolates grouped into two distinctly separate lineages with majority of the Indian isolates in Arctic like 1 lineage and the remaining isolates in sub-continental lineage. Isolates of the two distinct lineages were identified simultaneously from the same geographical region. Time scaled phylogenetic tree indicated that the sub-continental lineage of the virus is one of the earliest clade of rabies virus that diverged from bat rabies virus. On the contrary, the Arctic-like 1 lineage of India appeared to be a more recent divergence event. The amino acid sequence comparison revealed that all the major antigenic sites were almost conserved among the Indian isolates whereas few amino acid variations could be identified around site IIa, minor site I and IV. The dN/dS study based on G ecto-domain is in support of the earlier reports of strong purifying selection. In conclusion, it is evident that the Indian rabies virus isolates are of two major distinct lineages with distant phylogenetic and evolutionary relationship. PMID:25077994

Reddy, R V Chandrasekhar; Mohana Subramanian, B; Surendra, K S N L; Babu, R P Aravindh; Rana, S K; Manjari, K Sunitha; Srinivasan, V A

2014-10-01

96

Adaptive evolution of simian immunodeficiency viruses isolated from two conventional progressor macaques with neuroaids  

SciTech Connect

Simian immunodeficiency virus infection of macaques may result in neuroAIDS, a feature more commonly observed in macaques with rapid progressive disease than in those with conventional disease. This is the first report of two conventional progressors (H631 and H636) with encephalitis in rhesus macaques inoculated with a derivative of SIVsmES43-3. Phylogenetic analyses of viruses isolated from the cerebral spinal fluid (CSF) and plasma from both animals demonstrated tissue compartmentalization. Additionally, virus from the central nervous system (CNS) was able to infect primary macaque monocyte-derived macrophages more efficiently than virus from plasma. Conversely, virus isolated from plasma was able to replicate better in peripheral blood mononuclear cells than virus from CNS. We speculate that these viruses were under different selective pressures in their separate compartments. Furthermore, these viruses appear to have undergone adaptive evolution to preferentially replicate in their respective cell targets. Analysis of the number of potential N-linked glycosylation sites (PNGS) in gp160 showed that there was a statistically significant loss of PNGS in viruses isolated from CNS in both macaques compared to SIVsmE543-3. Moreover, virus isolated from the brain in H631, had statistically significant loss of PNGS compared to virus isolated from CSF and plasma of the same animal. It is possible that the brain isolate may have adapted to decrease the number of PNGS given that humoral immune selection pressure is less likely to be encountered in the brain. These viruses provide a relevant model to study the adaptations required for SIV to induce encephalitis.

Foley, Brian T [Los Alamos National Laboratory; Korber, Bette T [Los Alamos National Laboratory

2008-01-01

97

Complete Genome Sequencing of Mosquito and Human Isolates of Ngari Virus  

PubMed Central

Ngari virus (NRIV) is a recently described, naturally occurring reassortant between two other orthobunyaviruses, Bunyamwera virus (BUNV) and Batai virus (BATV). Intriguingly, this reassortment was associated with the acquisition of heightened virulence, although the molecular basis for this is not understood. Here we report the first complete genome sequences of Ngari virus. We include five isolates from various geographical locations, as well as samples isolated from both mosquitos and human cases. Based on an analysis of these sequence data, NRIVs are clearly genetically distinct from all known BUNV and BATV strains but are very closely related to one another regardless of their source. PMID:23166252

Groseth, Allison; Weisend, Carla

2012-01-01

98

Complete genome sequencing of mosquito and human isolates of Ngari virus.  

PubMed

Ngari virus (NRIV) is a recently described, naturally occurring reassortant between two other orthobunyaviruses, Bunyamwera virus (BUNV) and Batai virus (BATV). Intriguingly, this reassortment was associated with the acquisition of heightened virulence, although the molecular basis for this is not understood. Here we report the first complete genome sequences of Ngari virus. We include five isolates from various geographical locations, as well as samples isolated from both mosquitos and human cases. Based on an analysis of these sequence data, NRIVs are clearly genetically distinct from all known BUNV and BATV strains but are very closely related to one another regardless of their source. PMID:23166252

Groseth, Allison; Weisend, Carla; Ebihara, Hideki

2012-12-01

99

Isolation of a phylogenetically distinct rabies virus from a tufted capuchin monkey (Cebus apella) in Brazil.  

PubMed

A rabies virus isolate (BRmk1358 strain) was discovered from a rabid tufted capuchin monkey in Brazil. The present study determined the nucleotide sequence of the BRmk1358 strain and compared with the rabies viruses isolated from marmosets and other animals in the Americas. Phylogenetic analyses showed that the BRmk1358 strain formed a lineage distant from that of marmoset rabies virus within the Chiroptera-related rabies virus cluster. This result suggests that the source of rabies infection in the tufted capuchin monkey may have been bat, and that they have a risk to act as rabies reservoir in Brazil. PMID:24055656

Kobayashi, Yuki; Sugimoto, Kahori; Mochizuki, Nobuyuki; Segawa, Takao; Itou, Takuya; Carvalho, Adolorata A B; Nociti, Darci P; Mello, Rosane M; Santos, Anna K R A; Ito, Fumio H; Sakai, Takeo

2013-12-26

100

Phylogenetic analysis of hepatitis E virus isolates from India (1976-1993)  

Microsoft Academic Search

Seventeen Indian hepatitis E virus (HEV) isolates, representing epidemic and sporadic hepatitis E cases during 1976-1991, were sequenced in the RNA polymerase (RNAP) region. Five isolates were also sequenced in the non-structural hypervariable region of open reading frame 1. Open reading frames 2 and 3 were sequenced only for the prototype isolate. On the basis of the comparison of all

V. A. Arankalle; S. Paranjape; S. U. Emerson; R. H. Purcell; A. M. Walimbe

101

Genetic and antigenic variability in bovine viral diarrhea virus (BVDV) isolates from Belgium  

Microsoft Academic Search

This report describes the genetic and antigenic variability of bovine viral diarrhea virus strains isolated in Belgium. Part of the 5? untranslated region and the 5? end of the gp53 (E2) coding sequence were amplified by PCR and sequenced. Phylogenetic analysis showed that most field isolates segregated into genotypes Ib or II. Only one out of 28 field isolates belonged

B. Couvreur; C. Letellier; A. Collard; P. Quenon; P. Dehan; C. Hamers; P.-P. Pastoret; P. Kerkhofs

2002-01-01

102

Isolation and characterization of viruses related to the SARS coronavirus from animals in southern China.  

PubMed

A novel coronavirus (SCoV) is the etiological agent of severe acute respiratory syndrome (SARS). SCoV-like viruses were isolated from Himalayan palm civets found in a live-animal market in Guangdong, China. Evidence of virus infection was also detected in other animals (including a raccoon dog, Nyctereutes procyonoides) and in humans working at the same market. All the animal isolates retain a 29-nucleotide sequence that is not found in most human isolates. The detection of SCoV-like viruses in small, live wild mammals in a retail market indicates a route of interspecies transmission, although the natural reservoir is not known. PMID:12958366

Guan, Y; Zheng, B J; He, Y Q; Liu, X L; Zhuang, Z X; Cheung, C L; Luo, S W; Li, P H; Zhang, L J; Guan, Y J; Butt, K M; Wong, K L; Chan, K W; Lim, W; Shortridge, K F; Yuen, K Y; Peiris, J S M; Poon, L L M

2003-10-10

103

Molecular characterization of low pathogenic avian influenza viruses, isolated from food products imported into Singapore.  

PubMed

We have completed the genetic characterization of all eight gene segments for four low pathogenic avian influenza (LPAI) viruses. The objective of this study was to detect the presence of novel signatures that may serve as early warning indicators of the conversion of LPAI viruses to high pathogenic avian influenza (HPAI) viruses. This study included three H5N2 and one H5N3 viruses that were isolated from live poultry imported into Singapore as part of the national avian influenza virus (AIV) surveillance program. Based on the molecular criterion of the World Organisation for Animal Health (OIE), sequence analysis with the translated amino acid (aa) sequence of the hemagglutinin (HA) gene revealed the absence of multibasic aa at the HA cleavage site, identifying all four virus isolates as LPAI. Detailed phylogenetic tree analyses using the HA and neuraminidase (NA) genes clustered these isolates in the Eurasian H5 lineage, but away from the HPAI H5 subtypes. This analysis further revealed that the internal genes clustered to different avian and swine subtypes, suggesting that the four isolates may possibly share their ancestry with these different influenza subtypes. Our results suggest that the four LPAI isolates in this study contained mainly avian signatures, and the phylogenetic tree for the internal genes further suggests the potential for reassortment with other different circulating avian subtypes. This is the first comprehensive report on the genetic characterization of LPAI H5N2/3 viruses isolated in South-East Asia. PMID:19464126

Yeo, Dawn Su-Yin; Ng, Sock-Hoon; Liaw, Chin-Wen; Ng, Ley-Moy; Wee, Eugene Jing-Hui; Lim, Elizabeth Ai-Sim; Seah, Shirely Lay-Kheng; Wong, Wai-Kwan; Lim, Chee-Wee; Sugrue, Richard J; Tan, Boon-Huan

2009-09-18

104

[Phylogenetic analysis of rabies viruses isolated from animals in Tokyo in the 1950s].  

PubMed

Molecular epidemiological analysis of 96 rabies viruses isolated from animals in Tokyo in the 1950s involves Japanese fixed virus, Komatsugawa, Takamen, and Nishigahara strains. Strains isolated in Tokyo were divided into Tokyo 1 and Tokyo 2, and grouped into a worldwide distribution cluster differing from Takamen and Nishigahara. Tokyo 1 was grouped into the same cluster as viruses isolated from United States west coast dogs in the 1930s and 1940s. Tokyo 2 was grouped into the same cluster as the Komatsugawa strain, also known as a cluster of viruses from the Khabarovsk raccoon dog, and the Lake Baikal stepped fox in Russia. These findings suggest that 1950s Tokyo rabies viruses were related to those in Russia and the USA. PMID:21706842

Hatakeyama, Kaoru; Sadamasu, Kenji; Kai, Akemi

2011-05-01

105

Dissemination of influenza virus between anatomically isolated sites in ferrets.  

PubMed

Influenza virus was shown to disseminate from the respiratory tract of nasally infected ferrets to a surgically formed, subcutaneous tracheal pouch. Conversely, ferrets infected via the pouch shed virus from the upper respiratory tract. These events occurred within the first 24 h after infection. Passively administered ferret antibody to virus did not prevent the dissemination. PMID:4816632

Barber, W H; Small, P A

1974-03-01

106

Dengue Virus Serotype 2 from a Sylvatic Lineage Isolated from a Patient with Dengue Hemorrhagic Fever  

PubMed Central

Dengue viruses circulate in both human and sylvatic cycles. Although dengue viruses (DENV) infecting humans can cause major epidemics and severe disease, relatively little is known about the epidemiology and etiology of sylvatic dengue viruses. A 20-year-old male developed dengue hemorrhagic fever (DHF) with thrombocytopenia (12,000/ul) and a raised hematocrit (29.5% above baseline) in January 2008 in Malaysia. Dengue virus serotype 2 was isolated from his blood on day 4 of fever. A phylogenetic analysis of the complete genome sequence revealed that this virus was a member of a sylvatic lineage of DENV-2 and most closely related to a virus isolated from a sentinel monkey in Malaysia in 1970. This is the first identification of a sylvatic DENV circulating in Asia since 1975. PMID:19399166

Cardosa, Jane; Ooi, Mong How; Tio, Phaik Hooi; Perera, David; Holmes, Edward C.; Bibi, Khatijar; Abdul Manap, Zahara

2009-01-01

107

Isolation of West Nile virus from mosquitoes, crows, and a Cooper's hawk in Connecticut.  

PubMed

West Nile (WN) virus, a mosquito-transmitted virus native to Africa, Asia, and Europe, was isolated from two species of mosquitoes, Culex pipiens and Aedes vexans, and from brain tissues of 28 American crows, Corvus brachyrhynchos, and one Cooper's hawk, Accipiter cooperii, in Connecticut. A portion of the genome of virus isolates from four different hosts was sequenced and analyzed by comparative phylogenetic analysis. Our isolates from Connecticut were similar to one another and most closely related to two WN isolates from Romania (2.8 and 3.6 percent difference). If established in North America, WN virus will likely have severe effects on human health and on the health of populations of birds. PMID:10600741

Anderson, J F; Andreadis, T G; Vossbrinck, C R; Tirrell, S; Wakem, E M; French, R A; Garmendia, A E; Van Kruiningen, H J

1999-12-17

108

Complete genome sequence of a Dengue virus serotype 4 strain isolated in Roraima, Brazil.  

PubMed

Dengue is the most important arboviral disease worldwide. We report the complete genome sequence of a dengue virus serotype 4, genotype II strain isolated in 2010 from a patient with classical dengue fever in Boa Vista, Roraima, Brazil. PMID:22247521

Naveca, Felipe G; Souza, Victor C; Silva, George A V; Maito, Rodrigo M; Granja, Fabiana; Siqueira, Thalita; Acosta, Pablo O A

2012-02-01

109

Molecular Epidemiologic Studies on North American H9 Avian Influenza Virus Isolates from Waterfowl and Shorebirds  

Microsoft Academic Search

sequence data on H9 avian influenza virus (AIV) from wild birds are currently limited, we set out to determine the sequence of the hemagglutinin (HA) gene of H9 viruses circulating in North American waterfowl and shorebirds. In this study, we examined the HA gene from H9 AIV isolated from mallards (Anas platyrhynchos) sampled during 1998 and 1999 in Minnesota and

Mark W. Jackwood; David E. Stallknecht

2006-01-01

110

Monoclonal antibodies with neutralizing activity segregate isolates of bovine viral diarrhea virus into groups  

Microsoft Academic Search

Summary Isolates of bovine viral diarrhea (BVD) virus were differentiated by monoclonal antibodies (MoAbs) reactive with the 56kD viral polypeptide. Patterns of neutralizing activity of the MoAbs indicate that multiple epitopes are involved in virus neutralization.

S. Bolin; V. Moennig; N. E. Kelso Gourley; J. Ridpath

1988-01-01

111

THE ISOLATION OF BOVINE EPHEMERAL FEVER VIRUS IN CELL CULTURES AND EVIDENCE FOR AUTOINTERFERENCE  

Microsoft Academic Search

Bovine ephemeral fever (BEF) virus was isolated from infected fresh cattle blood directly into Vero cell cultures. One cycle of rapid freezethaw destroyed the infectivity of the virus to Vero cells. The infectivity to baby mice by intracerebral inoculation, on the other hand, was only reduced. The occurrence of autointerferencc due to the presence of defective interfering particles in cell

S Tzipori

1975-01-01

112

VECTOR-BORNE DISEASES, SURVEILLANCE, PREVENTION Isolations of Cache Valley Virus From Aedes albopictus (Diptera  

E-print Network

transcription-polymerase chain reaction assays. Three virus isolates were obtained from 34,567 Ă?eld-collected Ae virus (WNV) by reverse transcription-polymerase chain reaction, and all were negative. These results in the Northeast only WNV has been de- tected in Ae. albopictus using polymerase chain reac- tion (PCR) based

113

Characterization of a natural Plum pox virus isolate bearing a truncated coat protein.  

PubMed

Plum pox virus (PPV) isolates were collected in Hungary from plum varieties. PCR targeting the 3' genomic region resulted in a shorter PCR product in the case of the B1298 isolate bearing a 135-nucleotide deletion in frame in the N-terminal part of the coat protein (CP). The isolate was aphid-transmissible and the virion diameter was reduced compared to PPV-SK68. Detectability of this isolate by Western blot varied according to the antibody used. Integration of the deleted CP gene into an infectious PPV clone had no effect on infectivity and symptomatology. In competition experiments, B1298 had a considerable advantage in virus accumulation. PMID:19082685

Szathmáry, Erzsébet; Nádudvari, Júlia Novák; Szabó, László; Tóbiás, István; Balázs, Ervin; Palkovics, László

2009-01-01

114

Nucleotide sequence and phylogenetic analysis of Newcastle disease virus isolates from recent outbreaks in Taiwan.  

PubMed

Portions of the hemagglutinin neuraminidase (HN) gene of Newcastle disease virus (NDV) isolates from two recent outbreaks were sequenced to investigate epidemiology of this disease in Taiwan. These NDV isolates were all viscerotropic velogenic according to the clinical lesions produced in chickens. Sequence data were obtained from 14 NDV isolates (12 from 1995 and 2 from 1984). All isolates differed in their nucleotide sequences (from 0.3 to 15.3%), and represented potentially different strains of NDV. Phylogenetic analysis revealed that these isolates are closely related to viruses isolated from Japan and Malaysia. Some viruses isolated in 1995 appeared to evolve from viruses isolated in 1984. The results suggest that the 1995 outbreak of Newcastle disease (ND) in Taiwan may have been caused by multiple strains of velogenic NDV that have cocirculated in Taiwan for some time. Moreover, NDV isolates from racing pigeons were very similar to isolates from chickens in the same period, suggesting that both domestic and free-living birds were involved in the spread of ND in Taiwan. PMID:9201401

Yang, C Y; Chang, P C; Hwang, J M; Shieh, H K

1997-01-01

115

Isolation and Transmission of Human Retrovirus (Human T-Cell Leukemia Virus)  

Microsoft Academic Search

Nine new isolates of human T-cell leukemia-lymphoma virus (HTLV) were obtained from cells of seven patients with malignancies of mature T cells and from two clinically normal relatives of a T-cell leukemia patient. These people were from the United States, Israel, the West Indies, and Japan. The virus was detected in the fresh T cells and was isolated from the

M. Popovic; P. S. Sarin; M. Robert-Gurroff; V. S. Kalyanaraman; D. Mann; J. Minowada; R. C. Gallo

1983-01-01

116

Isolation methods and electron microscopy of the Internal Cork Virus of sweet potatoes  

E-print Network

ISOLATION METHODS AND ELECTRON MICROSCOPY OF THE INTERNAL CORK VIRUS OF SWEET POTATOES A Thesis By Edgar Eugene Pickens Submitted to the Graduate College of the Texas A8cM University in partial fulfillment of the requirements for the degree... of MASTER OF SCIENCE August 1967 Major Subject Biochemistry ISOLATION METHODS AND ELECTRON MICROSCOPY OF THE INTERNAL CORK VIRUS OF SWEET POTATOES A Thesis Edgar Eugene Pickens Approved as to style and content by: (Cnairman of Committee) (Head wf...

Pickens, Edgar Eugene

2012-06-07

117

Susceptibility of different plant species and tomato cultivars to two isolates of Pepino mosaic virus  

Microsoft Academic Search

As Pepino mosaic virus has become a pathogen of major importance in worldwide tomato production, information is needed on possible differences between\\u000a the sensitivity of cultivars towards infection. Furthermore, it is important what hosts other than Solanaceae may be virus reservoirs and are, therefore, threats for tomato cultivation. Two PepMV isolates (PepMV-Sav, E397, a European\\u000a tomato isolate and PV-0554, a

Ahmad Fakhro; Susanne von Bargen; Martina Bandte; Carmen Büttner; Philipp Franken; Dietmar Schwarz

2011-01-01

118

Comparative Study of Influenza Virus H2 (Asian) Hemagglutinins Isolated from Human and Avian Sources  

Microsoft Academic Search

Summary The hemagglutinin of an influenza virus isolated from a wild duck (Pintail, Anas acutd) in the USSR in 1976 had been found to be antigenically indistinguishable from the hemagglutinin of H2N2 viruses of human origin isolated in 1957. The hemagglutinins from viral preparations of the A\\/Anas acuta\\/Primorie\\/695\\/76 (H2Nav2) and A\\/Singapore\\/1\\/57 (H2N2) strains were purified by SDS gel chromatography as

Doris J. Bucher; Igor G. Kharitonenkov; Dema K. Lvov; Tamara V. Pysina; How-Ming Lee

1980-01-01

119

Genetic analysis of influenza B viruses isolated in Uganda during the 2009-2010 seasons  

PubMed Central

Background Influenza B viruses can cause morbidity and mortality in humans but due to the lack of an animal reservoir are not associated with pandemics. Because of this, there is relatively limited genetic sequences available for influenza B viruses, especially from developing countries. Complete genome analysis of one influenza B virus and several gene segments of other influenza B viruses isolated from Uganda from May 2009 through December 2010 was therefore undertaken in this study. Methods Samples were collected from patients showing influenza like illness and screened for influenza A and B by PCR. Influenza B viruses were isolated on Madin-Darby Canine Kidney cells and selected isolates were subsequently sequenced and analyzed phylogenetically. Findings Of the 2,089 samples collected during the period, 292 were positive by PCR for influenza A or B; 12.3% of the PCR positives were influenza B. Thirty influenza B viruses were recovered and of these 25 that grew well consistently on subculture were subjected to further analysis. All the isolates belonged to the B/Victoria-lineage as identified by hemagglutination inhibition assay and genetic analysis except one isolate that grouped with the B-Yamagata-lineage. The Ugandan B/Victoria-lineage isolates grouped in clade 1 which was defined by the N75K, N165K and S172P substitutions in hemagglutinin (HA) protein clustered together with the B/Brisbane/60/2008 vaccine strain. The Yamagata-like Ugandan strain, B/Uganda/MUWRP-053/2009, clustered with clade 3 Yamagata viruses such as B/Bangladesh/3333/2007 which is characterized by S150I and N166Y substitutions in HA. Conclusion In general there was limited variation among the Ugandan isolates but they were interestingly closer to viruses from West and North Africa than from neighboring Kenya. Our isolates closely matched the World Health Organization recommended vaccines for the seasons. PMID:23289789

2013-01-01

120

Autochthonous Hepatitis E Virus Infection in Germany with Sequence Similarities to Other European Isolates  

Microsoft Academic Search

Hepatitis E virus (HEV) is a common cause of acute hepatitis in endemic areas. Yet reports on autochthonous cases in other\\u000a areas such as middle Europe are increasing. Here we report on a patient, who obviously acquired his HEV infection in Germany.\\u000a Sequence analysis of the virus gained from his serum revealed homologies to other European isolates and swine isolates.

J. C. Preiss; A. Plentz; E. Engelmann; T. Schneider; W. Jilg; M. Zeitz; R. Duchmann

2006-01-01

121

Molecular differentiation of cytopathic and noncytopathic isolates of Bovine Viral Diarrhea Virus  

E-print Network

MOLECULAR DIFFERENTIATION OF CYTOPATHIC AND NONCYTOPATHIC ISOLATES OF BOVINE VIRAL DIARRHEA VIRUS A Thesis by LYNDA LEDAWN BISSEY Submitted to the Office of Graduate Studies of Texas A&M University in partial fulfillment of requirements... for the degree of MASTER OF SCIENCE August 1989 Major Subject: Veterinary Microbiology MOLECULAR DIFFERENTIATION OF CYTOPATHIC AND NONCYTOPATHIC ISOLATES OF BOVINE VIRAL DIARRHEA VIRUS A Thesis by LYNDA LEDAWN BISSEY Approved as to style and content by...

Bissey, Lynda LeDawn

2012-06-07

122

Isolation and mutation trend analysis of influenza A virus subtype H9N2 in Egypt  

PubMed Central

Background Avian influenza virus H9N2 is a panzootic pathogen that affects poultry causing mild to moderate respiratory distress but has been associated with high morbidity and considerable mortality. Interspecies transmission of H9N2 from avian species to mammalian hosts does occur. The virus possesses human virus-like receptor specificity and it can infect humans producing flu-like illness. Methods Recently, mild influenza like symptoms were detected in H5N1 vaccinated flocks. Influenza A subtype H9N2 was isolated from the infected flock. The virus evolution was investigated by sequencing the viral genes to screen the possible virus recombination. The viral amino acid sequences from the isolated H9N2 strains were compared to other related sequences from the flu data base that were used to assess the robustness of the mutation trend. Changes in the species-associated amino acid residues or those that enabled virulence to mammals were allocated. Results Phylogenetic analyses of haemagglutinin and neuraminidase genes showed that the recently isolated Egyptian strain belonged to the H9N2 sub-lineage that prevails in Israel. The six internal segments of the isolated virus were found to be derived from the same sub-lineage with no new evidence of reassortment. The results demonstrated conserved genetic and biological constitution of H9N2 viruses in the Middle East. The recently isolated H9N2 virus from chicken in Egypt possessed amino acids that could enable the virus to replicate in mammals and caused severe disease in domestic chickens. Conclusion The study highlights the importance of continuous monitoring of the mutations evolved in avian influenza viruses and its impact on virulence to avian species in addition to its importance in the emergence of new strains with the capacity to be a pandemic candidate. PMID:22925485

2012-01-01

123

Pathogenesis of Newcastle disease in vaccinated chickens: pathogenicity of isolated virus and vaccine effect on challenge of its virus.  

PubMed

The pathogenicity of Newcastle disease (ND) virus, isolated from ND outbreak in vaccinated chickens, was evaluated through experiments. The pathogenicity indexes (mean death time (MDT); 58 hr, intracerebral pathogenicity index (ICPI); 1.7 and intravenous pathogenicity index (IVPI); 2.51) indicated that the ND virus was velogenic. The ND virus caused lymphocytic necrosis in the spleen with fibrinous exudation and proliferation of macrophages, sinusoidal fibrin exudation in the liver, proliferation of macrophages in the lung, lymphocytic necrosis and depletion in the bursa of Fabricius, cecal tonsils and thymus, necrosis of bone marrow, tracheitis, conjunctivitis and necrosis of feather epithelial cells in specific-pathogen-free chickens. Immunohistochemically, ND virus antigens were seen in the lesions mentioned above. The ND virus could not induce the encephalitis and pancreatitis that were observed in the natural case of ND in vaccinated chickens. There was no clinical disease in vaccinated chickens after the challenge of the ND virus. In diluted ND vaccine experiments, chickens vaccinated with a high dilution of vaccine and then challenged with the ND virus showed clinical sign and mortality with pancreatic focal necrosis. Vaccine diluted with fresh tap water had no effect on protection against the challenge of the ND virus. This study suggests that improper vaccination may be involved in outbreaks of ND in vaccinated chickens. PMID:23966012

Nakamura, Kikuyasu; Ito, Mitsuru; Nakamura, Toshiki; Yamamoto, Yu; Yamada, Manabu; Mase, Masaji; Imai, Kunitoshi

2014-01-01

124

RNA interference inhibits herpes simplex virus type 1 isolated from saliva samples and mucocutaneous lesions.  

PubMed

The aim of this study was to evaluate the use of RNA interference to inhibit herpes simplex virus type-1 replication in vitro. For herpes simplex virus type-1 gene silencing, three different small interfering RNAs (siRNAs) targeting the herpes simplex virus type-1 UL39 gene (sequence si-UL 39-1, si-UL 39-2, and si-UL 39-3) were used, which encode the large subunit of ribonucleotide reductase, an essential enzyme for DNA synthesis. Herpes simplex virus type-1 was isolated from saliva samples and mucocutaneous lesions from infected patients. All mucocutaneous lesions' samples were positive for herpes simplex virus type-1 by real-time PCR and by virus isolation; all herpes simplex virus type-1 from saliva samples were positive by real-time PCR and 50% were positive by virus isolation. The levels of herpes simplex virus type-1 DNA remaining after siRNA treatment were assessed by real-time PCR, whose results demonstrated that the effect of siRNAs on gene expression depends on siRNA concentration. The three siRNA sequences used were able to inhibit viral replication, assessed by real-time PCR and plaque assays and among them, the sequence si-UL 39-1 was the most effective. This sequence inhibited 99% of herpes simplex virus type-1 replication. The results demonstrate that silencing herpes simplex virus type-1 UL39 expression by siRNAs effectively inhibits herpes simplex virus type-1 replication, suggesting that siRNA based antiviral strategy may be a potential therapeutic alternative. PMID:24835621

Silva, Amanda Perse da; Lopes, Juliana Freitas; Paula, Vanessa Salete de

2014-01-01

125

Influenza A (H15N4) Virus Isolation in Western Siberia, Russia  

PubMed Central

The rarely identified influenza A viruses of the H15 hemagglutinin subtype have been isolated exclusively in Australia. Here we report the isolation of an H15N4 influenza A virus (A/teal/Chany/7119/2008) in Western Siberia, Russia. Phylogenetic analysis demonstrated that the internal genes of the A/teal/Chany/7119/2008 strain belong to the Eurasian clade and that the H15 and N4 genes were introduced into the gene pool of circulating endemic avian influenza viruses through reassortment events. PMID:23283950

Sivay, Mariya V.; Baranovich, Tatiana; Marchenko, Vasiliy Y.; Sharshov, Kirill A.; Govorkova, Elena A.; Shestopalov, Aleksander M.

2013-01-01

126

West Nile virus isolates from mosquitoes in New York and New Jersey, 1999.  

PubMed Central

An outbreak of encephalitis due to West Nile (WN) virus occurred in New York City and the surrounding areas during 1999. Mosquitoes were collected as part of a comprehensive surveillance program implemented to monitor the outbreak. More than 32,000 mosquitoes representing 24 species were tested, and 15 WN virus isolates were obtained. Molecular techniques were used to identify the species represented in the WN virus-positive mosquito pools. Most isolates were from pools containing Culex pipiens mosquitoes, but several pools contained two or more Culex species. PMID:11585523

Nasci, R. S.; White, D. J.; Stirling, H.; Oliver, J. A.; Daniels, T. J.; Falco, R. C.; Campbell, S.; Crans, W. J.; Savage, H. M.; Lanciotti, R. S.; Moore, C. G.; Godsey, M. S.; Gottfried, K. L.; Mitchell, C. J.

2001-01-01

127

Isolation and full genomic characterization of Batai virus from mosquitoes, Italy 2009.  

PubMed

In 2009, 2589 mosquitoes were collected in northwest Italy and screened for orthobunyavirus RNA by RT-PCR. One pool of Anopheles maculipennis complex mosquitoes was found to be positive and a virus was isolated from that pool. The isolate was identified as Batai virus (BATV) by sequencing. Previously, BATV was detected in Italy, but limited data and no prior isolates existed. Full-length sequences of the S, M and L segments were determined for the newly isolated Italian strain. For comparison, partial sequences were also determined for the BATV strain Calovo (former Czechoslovakia, 1960). Phylogenetic analyses revealed clustering of the newly derived Italian BATV along with a recent isolate from Germany and the historic strain Calovo. To the best of our knowledge, this represents the first isolation of BATV from Italy, which confirms a broader geographical distribution of BATV in Europe than was previously verified by isolation. PMID:23515020

Huhtamo, Eili; Lambert, Amy J; Costantino, Stefano; Servino, Luca; Krizmancic, Letizia; Boldorini, Renzo; Allegrini, Sara; Grasso, Ivan; Korhonen, Essi M; Vapalahti, Olli; Lanciotti, Robert S; Ravanini, Paolo

2013-06-01

128

Modoc-like virus isolated from wild deer mice (Peromyscus maniculatus) in Alberta.  

PubMed

Small mammals were trapped in northeastern Alberta, Canada during 1976. Blood samples from these animals were tested for virus by inoculation of suckling mice. Blood clots from two deer mice yielded isolates of the same virus. The virus was related antigenically to a number of flaviviruses which have been isolated from mammals in Central America and North America and was related most closely to Modoc virus. Physical, chemical, and biological properties of the virus were similar also to those of Modoc virus. It did not produce illness or death in deer mice inoculated in the laboratory. Neutralization tests indicated that 1/38 (3%) red squirrels (Tamiasciurus hudsonicus), 3/35 (9%) least chipmunks (Eutamius minimus), 13/109 (12%) deer mice, and 3/50 (6%) humans were infected naturally. This is the first reported evidence of infection of red squirrels and chipmunks with a Modoc-like virus. These data extend the range of Modoc-like viruses northward by 1,500 km and comprise the first isolate from mammals in the boreal forest of Canada. PMID:2987550

Zarnke, R L; Yuill, T M

1985-04-01

129

Biological and Genomic Sequence Characterization of Maize streak virus Isolates from Wheat.  

PubMed

ABSTRACT Maize streak virus (MSV) is best known as the causal agent of maize streak disease. However, only a genetically uniform subset of the viruses within this diverse species is actually capable of producing severe symptoms in maize. Whereas these "maize-type" viruses all share greater than 95% sequence identity, MSV strains isolated from grasses may share as little as 79% sequence identity with the maize-type viruses. Here, we present the complete genome sequences and biological characterization of two MSV isolates from wheat that share approximately 89% sequence identity with the maize-type viruses. Clonal populations of these two isolates, named MSV-Tas and MSV-VW, were leafhopper-transmitted to Digitaria sanguinalis and a range of maize, wheat, and barley genotypes. Whereas the two viruses showed some differences in their pathogenicity in maize, they were both equally pathogenic in D. sanguinalis and the various wheat and barley genotypes tested. Phylogenetic analyses involving the genome sequences of MSV-Tas and MSV-VW, a new maize-type virus also fully sequenced in this study (MSV-VM), and all other available African streak virus sequences, indicated that MSV-Tas and MSV-VW are close relatives that together represent a distinct MSV strain. Sequence analyses revealed that MSV-VM has a recombinant genome containing MSV-Tas/VW-like sequences within its movement protein gene. PMID:18944143

Willment, J A; Martin, D P; Van der Walt, E; Rybicki, E P

2002-01-01

130

Genetic characterization of a noncytopathic bovine viral diarrhea virus 2b isolated from cattle in China.  

PubMed

In January 2013, several clinical signs of cattle with diarrhea, cough, nasal discharge, and fever were reported in Jilin province, China. One virus named SD1301 was isolated and identified. Complete genome of the virus is 12258nt in length and contains a 5'UTR, one open reading frame encoding a polyprotein of 3,897 amino acids and a 3'UTR. Phylogenetic analysis of 5'UTR, N(pro), E1 and E2 gene demonstrated the virus belonged to BVDV 2b, and genetically related to the BVDV strain Hokudai-Lab/09 from Japan in 2010. This bovine viral diarrhea virus displays a unique genetic signature with 27-nucleotide deletion in the 5'UTR, which is similar to the bovine viral diarrhea virus C413 (AF002227). This was the first confirmed isolation of ncp BVDV2b circulating in bovine herd of China. PMID:24811746

Wang, Wei; Shi, Xinchuan; Chen, Chaoyang; Wu, Hua

2014-10-01

131

Respiratory transmission of an avian H3N8 influenza virus isolated from a harbour seal  

USGS Publications Warehouse

The ongoing human H7N9 influenza infections highlight the threat of emerging avian influenza viruses. In 2011, an avian H3N8 influenza virus isolated from moribund New England harbour seals was shown to have naturally acquired mutations known to increase the transmissibility of highly pathogenic H5N1 influenza viruses. To elucidate the potential human health threat, here we evaluate a panel of avian H3N8 viruses and find that the harbour seal virus displays increased affinity for mammalian receptors, transmits via respiratory droplets in ferrets and replicates in human lung cells. Analysis of a panel of human sera for H3N8 neutralizing antibodies suggests that there is no population-wide immunity to these viruses. The prevalence of H3N8 viruses in birds and multiple mammalian species including recent isolations from pigs and evidence that it was a past human pandemic virus make the need for surveillance and risk analysis of these viruses of public health importance.

Karlsson, Erik A.; Ip, Hon S.; Hall, Jeffrey S.; Yoon, Sun W.; Johnson, Jordan; Beck, Melinda A.; Webby, Richard J.; Schultz-Cherry, Stacey

2014-01-01

132

Respiratory transmission of an avian H3N8 influenza virus isolated from a harbour seal.  

PubMed

The ongoing human H7N9 influenza infections highlight the threat of emerging avian influenza viruses. In 2011, an avian H3N8 influenza virus isolated from moribund New England harbour seals was shown to have naturally acquired mutations known to increase the transmissibility of highly pathogenic H5N1 influenza viruses. To elucidate the potential human health threat, here we evaluate a panel of avian H3N8 viruses and find that the harbour seal virus displays increased affinity for mammalian receptors, transmits via respiratory droplets in ferrets and replicates in human lung cells. Analysis of a panel of human sera for H3N8 neutralizing antibodies suggests that there is no population-wide immunity to these viruses. The prevalence of H3N8 viruses in birds and multiple mammalian species including recent isolations from pigs and evidence that it was a past human pandemic virus make the need for surveillance and risk analysis of these viruses of public health importance. PMID:25183346

Karlsson, Erik A; Ip, Hon S; Hall, Jeffrey S; Yoon, Sun Woo; Johnson, Jordan; Beck, Melinda A; Webby, Richard J; Schultz-Cherry, Stacey

2014-01-01

133

Molecular characterization of a hepatitis E virus isolate from Namibia  

Microsoft Academic Search

Hepatitis E virus (HEV) causes sporadic and epidemic acute viral hepatitis in many developing countries. In Africa, hepatitis E has been documented by virus detection (reverse transcriptase polymerase chain reaction, RT-PCR) in Egypt, Chad, Algeria, Morocco and Tunisia. Cases of presumptive hepatitis E also have been documented by detection of antibody to HEV in the Sudan, Kenya, Ethiopia, Somalia, Djibouti

Junkun He; Leonard N. Binn; Sergei A. Tsarev; Curtis G. Hayes; John A. Frean; Margaretha Isaacson; Bruce L. Innis

2000-01-01

134

[First isolation of quang binh-like virus from mosquitoes in China].  

PubMed

This study aims to investigate the distribution patterns of mosquito-borne viruses in Menghai County, Xishuangbanna Prefecture, Yunnan Province, China and to provide evidence for the prevention and control of mosquito-borne diseases. Mosquito samples were collected using mosquito lamps. Viruses were isolated from the samples by cell culture, and the isolates were identified by RT-PCR. The genomes of isolates were sequenced for phylogenetic analysis. In July 2012, a total of 1468 mosquitoes were captured in Daluo Town of Menghai County; they were divided into 32 pools, including Culex tritaeniorhynchus (28 pools, 1383 mosquitoes), Culex quinquefasciatus (2 pools, 66 mosquitoes), and Anopheles (2 pools, 19 mosquitoes). Golden hamster kidney cells (BHK-21) and Aedes albopictus cells (C6/36) were used for virus isolation. The results showed that C6/36 cells were susceptible to two isolates recovered from Culex tritaeniorhynchus (BNDL1205 and BNDL1227), with marked cytopathic effect (CPE) of cell fusion. By contrast, the two isolates could not cause CPE in BHK-21 cells. RT-PCR was performed for the two isolates using the flavivirus-specific primers FU2/cFD3, and a 800-bp amplicon was obtained from both of them. Phylogenetic analysis showed that the two isolates shared the same evolutionary branch with the Quang Binh virus (QBV) strain VN180, which had been isolated from Vietnam, with nucleotide sequence homologies of 83.4% and 82.9%, respectively. However, there existed relatively large differences in nucleotide sequence between them and other Culex flavivirus strains previously isolated in China and other regions. In light of the similarity between the two isolates and QBV, BNDL1205 and BNDL122 were referred to as Quang Binh-like virus, which were first reported in China. PMID:24772899

Feng, Yun; Li, Hong-Bin; Zhu, Jin; Zhang, Yu-Zhen; Yang, Wei-Hong; Fan, Jian-Hua; Liang, Guo-Dong; Zhang, Hai-Lin

2014-01-01

135

Complete Genome Sequence of a New H9N2 Avian Influenza Virus Isolated in China  

PubMed Central

The complete genomic sequence of a new H9N2 avian influenza virus (AIV), isolated in northwestern China, was determined. Sequence and phylogenetic analyses based on the sequences of eight genomic segments revealed that the isolate is phylogenetically related to the Y280-like sublineage. PMID:23723395

Wang, Jing-Yu; Ren, Juan-Juan; Liu, Wan-Hua; Tang, Pan; Wu, Ning; Wang, Chi-Young; Chang, Ching-Dong

2013-01-01

136

Punique virus, a novel phlebovirus, related to sandfly fever Naples virus, isolated from sandflies collected in Tunisia  

PubMed Central

Sandflies are widely distributed around the Mediterranean Basin. Therefore, human populations in this area are potentially exposed to sandfly-transmitted diseases, including those caused by phleboviruses. Whilst there are substantial data in countries located in the northern part of the Mediterranean basin, few data are available for North Africa. In this study, a total of 1489 sandflies were collected in 2008 in Tunisia from two sites, bioclimatically distinct, located 235 km apart, and identified morphologically. Sandfly species comprised Phlebotomus perniciosus (52.2 %), Phlebotomus longicuspis (30.1 %), Phlebotomus papatasi (12 .0%), Phlebotomus perfiliewi (4.6 %), Phlebotomus langeroni (0.4 %) and Sergentomyia minuta (0.5 %). PCR screening, using generic primers for the genus Phlebovirus, resulted in the detection of ten positive pools. Sequence analysis revealed that two pools contained viral RNA corresponding to a novel virus closely related to sandfly fever Naples virus. Virus isolation in Vero cells was achieved from one pool. Genetic and phylogenetic characterization based on sequences in the three genomic segments showed that it was a novel virus distinct from other recognized members of the species. This novel virus was provisionally named Punique virus. Viral sequences in the polymerase gene corresponding to another phlebovirus closely related to but distinct from sandfly fever Sicilian virus were obtained from the eight remaining positive pools. PMID:20089800

Zhioua, Elyes; Moureau, Gregory; Chelbi, Ifhem; Ninove, Laetitia; Bichaud, Laurence; Derbali, Mohamed; Champs, Mylene; Cherni, Saifeddine; Salez, Nicolas; Cook, Shelley; de Lamballerie, Xavier; Charrel, Remi N.

2012-01-01

137

Complete genome and clinicopathological characterization of a virulent Newcastle disease virus isolate from South America.  

PubMed

Newcastle disease (ND) is one of the most important diseases of poultry, negatively affecting poultry production worldwide. The disease is caused by Newcastle disease virus (NDV) or avian paramyxovirus type 1 (APMV-1), a negative-sense single-stranded RNA virus of the genus Avulavirus, family Paramyxoviridae. Although all NDV isolates characterized to date belong to a single serotype of APMV-1, significant genetic diversity has been described between different NDV isolates. Here we present the complete genome sequence and the clinicopathological characterization of a virulent Newcastle disease virus isolate (NDV-Peru/08) obtained from poultry during an outbreak of ND in Peru in 2008. Phylogenetic reconstruction and analysis of the evolutionary distances between NDV-Peru/08 and other isolates representing established NDV genotypes revealed the existence of large genomic and amino differences that clearly distinguish this isolate from viruses of typical NDV genotypes. Although NDV-Peru/08 is a genetically distinct virus, pathogenesis studies conducted with chickens revealed that NDV-Peru/08 infection results in clinical signs characteristic of velogenic viscerotropic NDV strains. Additionally, vaccination studies have shown that an inactivated NDV-LaSota/46 vaccine conferred full protection from NDV-Peru/08-induced clinical disease and mortality. This represents the first complete characterization of a virulent NDV isolate from South America. PMID:22135263

Diel, Diego G; Susta, Leonardo; Cardenas Garcia, Stivalis; Killian, Mary L; Brown, Corrie C; Miller, Patti J; Afonso, Claudio L

2012-02-01

138

Chemokine coreceptor usage by diverse primary isolates of human immunodeficiency virus type 1.  

PubMed

We tested chemokine receptor subset usage by diverse, well-characterized primary viruses isolated from peripheral blood by monitoring viral replication with CCR1, CCR2b, CCR3, CCR5, and CXCR4 U87MG.CD4 transformed cell lines and STRL33/BONZO/TYMSTR and GPR15/BOB HOS.CD4 transformed cell lines. Primary viruses were isolated from 79 men with confirmed human immunodeficiency virus type 1 (HIV-1) infection from the Chicago component of the Multicenter AIDS Cohort Study at interval time points. Thirty-five additional well-characterized primary viruses representing HIV-1 group M subtypes A, B, C, D, and E and group O and three primary simian immunodeficiency virus (SIV) isolates were also used for these studies. The restricted use of the CCR5 chemokine receptor for viral entry was associated with infection by a virus having a non-syncytium-inducing phenotype and correlated with a reduced rate of disease progression and a prolonged disease-free interval. Conversely, broadening chemokine receptor usage from CCR5 to both CCR5 and CXCR4 was associated with infection by a virus having a syncytium-inducing phenotype and correlated with a faster rate of CD4 T-cell decline and progression of disease. We also observed a greater tendency for infection with a virus having a syncytium-inducing phenotype in men heterozygous for the defective CCR5 Delta32 allele (25%) than in those men homozygous for the wild-type CCR5 allele (6%) (P = 0.03). The propensity for infection with a virus having a syncytium-inducing phenotype provides a partial explanation for the rapid disease progression among some men heterozygous for the defective CCR5 Delta32 allele. Furthermore, we did not identify any primary viruses that used CCR3 as an entry cofactor, despite this CC chemokine receptor being expressed on the cell surface at a level commensurate with or higher than that observed for primary peripheral blood mononuclear cells. Whereas isolates of primary viruses of SIV also used STRL33/BONZO/TYMSTR and GPR15/BOB, no primary isolates of HIV-1 used these particular chemokine receptor-like orphan molecules as entry cofactors, suggesting a limited contribution of these other chemokine receptors to viral evolution. Thus, despite the number of chemokine receptors implicated in viral entry, CCR5 and CXCR4 are likely to be the physiologically relevant chemokine receptors used as entry cofactors in vivo by diverse strains of primary viruses isolated from blood. PMID:9765480

Zhang, L; He, T; Huang, Y; Chen, Z; Guo, Y; Wu, S; Kunstman, K J; Brown, R C; Phair, J P; Neumann, A U; Ho, D D; Wolinsky, S M

1998-11-01

139

Isolation of infectious pancreatic necrosis virus (serotype Ab) from diverse species of estuarine fish  

NASA Astrophysics Data System (ADS)

Two significant fish kills occurred in the Pamlico River estuary (North Carolina, USA), one in December 1981 and January 1982, and the other in June 1982. The first involved only the southern flounder (Paralichthys lethostigma). Histopathologic examination of morbid and moribund flounder revealed extensive sloughing and necrosis of the mucosa of the pyloric caeca and intestine, and inflammation of the submucosa of the pyloric caeca. Brain and internal organ homogenates from morbid and moribund flounder were assayed on CHSE-214 cells, and a virus was isolated. Virus titers ranged from?8.4 · 102 to 6.3 · 107 TCID50 per gram of tissue. Cross-plaque neutralization assays indicated that the southern flounder virus was infectious pancreatic necrosis virus serotype Ab. Immersion challenge showed the isolate is only slightly virulent for fry of brook trout (Salvelinus fontinalis). The second fish kill involved the southern flounder and six other species: hogchoker (Trinectes maculatus), Atlantic silverside (Menidia menidia), spot (Leiostomus xanthurus), Atlantic croaker (Micropogon undulatus), silver perch (Bairdiella chrysura), and striped mullet (Mugil cephalus). Virus was isolated from southern founder, hogchoker, Atlantic silverside, and spot. Neutralization assays indicated that the four isolates were nearly identical; however, the diversity of species affected suggests that the virus might not have been the specific cause of mortality.

McAllister, P. E.; Newman, M. W.; Sauber, J. H.; Owens, W. J.

1984-03-01

140

Genetic heterogeneity among isolates of Ross River virus from different geographical regions.  

PubMed Central

The RNase T1 maps of 80 isolates of Ross River virus from different regions of mainland Australia and the Pacific Islands were compared. Four different clusters of isolates with greater than an estimated 5 to 6% diversity at the nucleotide level were found. There was a pattern of differences between eastern and western Australian strains; however, the pattern was disturbed by overlaps and incursants. Pacific Islands isolates belonged to the eastern Australian topotype. Our findings suggest that certain genetic types of Ross River virus predominate in different geographical regions. In contrast, populations of other important Australian arboviruses (Murray Valley encephalitis, Kunjin, and Sindbis viruses) are distributed across the Australian continent as minor variants of one strain. Our data also show that in one region, strains of Ross River virus with identical RNase T1 maps circulate during both years when epidemics occur and years when they do not. This finding suggests that Ross River virus epidemics are not dependent on the introduction or evolution of new strains of the virus. Two strains, belonging to the eastern Australian topotype, were isolated in Western Australia. It is likely that viremic humans or possibly domestic livestock travelling by aircraft were responsible for this movement. Images PMID:8497065

Lindsay, M D; Coelen, R J; Mackenzie, J S

1993-01-01

141

Genetic characterization and pathogenicity assessment of Newcastle disease virus isolated from wild peacock.  

PubMed

The continued spread and occurrence of Newcastle disease virus (NDV) has posed potential threat to domestic poultry industry around the globe. Mainly, wild avian species has always been implicated for the natural reservoir for virus and spread of the disease. In the present study, we report the isolation of Newcastle disease virus (NDV/Peacock/India/2012) in necropsy brain tissue sample of wild peacock from North India. Complete genome of the virus was found to be 15,186 nucleotides (nts) with six genes in order of 3'-N-P-M-F-HN-L-5', which was limited by 55-nts leader region at the 3' end and a 114-nts trailer sequence at 5' end. Sequence analysis of fusion protein revealed the dibasic amino acid cleavage site (112)R-R-Q-K-R-F(117), a characteristic motif of virulent virus. Phylogenetic analysis placed the isolate in genotype II of Newcastle disease virus showing the lowest mean percent divergence (6 %) with other genotype II counterparts. The isolate was characterized as mesogenic (intermediate pathotype) based on the mean death time (63 h) in embryonated chicken eggs and the intra-cerebral pathogenicity index (1.40) in day-old chicks. The report emphasizes the dynamic ecology of NDV strains circulating in a wild avian host during the outbreak of 2012 in North India. Further the genotypic and pathotypical characterizations of the isolate could help in development of homologous vaccine against NDV strain circulating in avian population. PMID:25260553

Khulape, Sagar A; Gaikwad, Satish S; Chellappa, Madhan Mohan; Mishra, Bishnu Prasad; Dey, Sohini

2014-12-01

142

Characterization of H5N1 influenza A viruses isolated from domestic green-winged teal.  

PubMed

Two avian influenza virus strains, A/domestic green-winged teal/Hunan/67/2005 (H5N1) (D-GWT/67) and A/domestic green-winged teal/Hunan/79/2005 (H5N1) (D-GWT/79), were isolated from healthy domestic green-winged teals (Anas crecca) in Hunan Province, South China. Genomic analysis showed that both isolates were reassortants. The hemagglutinin (HA) genes of the two isolates were closely related to that of an H5N1 strain isolated from tree sparrow (A/tree sparrow/Henan/1/04). The neuraminidase (NA) genes and the internal protein genes of both isolates were closely related to those from A/chicken/Shantou/4231/2003-like (H5N1) viruses, with exception of the matrix (M) gene of D-GWT/79, which was closely related to that of the H7N3 strain A/mallard/Netherlands/12/2000 isolated from wild mallard duck. The virulence of the two isolates was examined in chickens, ducks, and mice. Both strains were found to be highly pathogenic in chickens and ducks, but showed low pathogenicity in mice. These findings contribute to the realization that domestic green-winged teals carrying the H5N1 virus may play an important role in transmitting the virus among birds. PMID:18825495

Chen, Jianjun; Yang, Zhongdong; Chen, Quanjiao; Liu, Xueying; Fang, Fang; Chang, Haiyan; Li, Dongmei; Chen, Ze

2009-02-01

143

Molecular epidemiologic studies on North American H9 avian influenza virus isolates from waterfowl and shorebirds.  

PubMed

Because sequence data on H9 avian influenza virus (AIV) from wild birds are currently limited, we set out to determine the sequence of the hemagglutinin (HA) gene of H9 viruses circulating in North American waterfowl and shorebirds. In this study, we examined the HA gene from H9 AIV isolated from mallards (Anas platyrhynchos) sampled during 1998 and 1999 in Minnesota and ruddy turnstones (Arenaria interpres) sampled during 2003 in Delaware and New Jersey. At these sites, the H9N2 subtype represented 12% and 4% of the avian influenza isolates from mallards in 1998 and 1999, respectively, and 8% of the AIVs isolated from shorebirds between 2000 and 2002. Sequences from these viruses were compared with sequences from H9 AIV isolated from commercial poultry and aquatic birds from North America, Europe, Asia, and the Middle East: four previously reported and three new clades were observed. Sequence data from the HA gene of North American waterfowl and shorebird isolates generated in this study most closely group with the Eurasian H9 viruses in the Y439 clade. In addition, the HA cleavage site (AASNR/G) and receptor binding site was identical to the representative virus of that group (DK/Hong Kong/Y439/97). Viruses in that clade are commonly found in ducks and chickens in Hong Kong and Korea. Positive evolutionary selection (dNonsynonymous > dSynonymous) was observed for the HA gene among the North American waterfowl and shorebird H9N2 viruses, indicating that the H9N2-type viruses are changing in their natural hosts. PMID:17494604

Jackwood, Mark W; Stallknecht, David E

2007-03-01

144

Sunguru virus: a novel virus in the family Rhabdoviridae isolated from a chicken in north-western Uganda.  

PubMed

Sunguru virus (SUNV), a novel virus belonging to the highly diverse Rhabdoviridae family, was isolated from a domestic chicken in the district of Arua, Uganda, in 2011. This is the first documented isolation of a rhabdovirus from a chicken. SUNV is related to, but distinct from, Boteke virus and other members of the unclassified Sandjimba group. The genome is 11056 nt in length and contains the five core rhabdovirus genes plus an additional C gene (within the ORF of a phosphoprotein gene) and a small hydrophobic protein (between the matrix and glycoprotein genes). Inoculation of vertebrate cells with SUNV resulted in significant viral growth, with a peak titre of 7.8 log10 p.f.u. ml(-1) observed in baby hamster kidney (BHK) cells. Little to no growth was observed in invertebrate cells and in live mosquitoes, with Anopheles gambiae mosquitoes having a 47.4% infection rate in the body but no dissemination of the virus to the salivary glands; this suggests that this novel virus is not arthropod borne as some other members of the family Rhabdoviridae. PMID:24718834

Ledermann, Jeremy P; Zeidner, Nord; Borland, Erin M; Mutebi, John-Paul; Lanciotti, Robert S; Miller, Barry R; Lutwama, Julius J; Tendo, Joseph M; Andama, Vincent; Powers, Ann M

2014-07-01

145

Comparison of different tissue cultures for isolation and quantitation of influenza and parainfluenza viruses.  

PubMed Central

Rhesus and cynomolgus monkey kidney tissue cultures and two continuous lines, Madin-Darby canine kidney (MDCK) and LLC-MK2, were compared in titrations and isolations of influenza and parainfluenza viruses. Tube cultures were inoculated with laboratory virus strains or stored patient specimens and observed for hemadsorption. Trypsin was added to the medium of the continuous lines to increase sensitivity. All four tissue cultures gave similar titers of influenza A/USSR (H1N1), A/Texas (H3N2), and B/HK, but lower titers of parainfluenza 1, 2, and 3 were observed with MDCK. Cynomolgus kidney was the best single tissue culture for reisolation of the six viruses, but foamy-virus contamination of many lots was a serious problem. Reisolation of influenza viruses was as successful with MDCK as with primary monkey kidney. LLC-MK2 was similar to rhesus kidney but less successful than cynomolgus kidney. For reisolation of parainfluenza viruses, LLC-MK2 was superior to rhesus monkey kidney and similar to cynomolgus kidney. MDCK was less useful for parainfluenza viruses. Thus, LLC-MK2 would be an acceptable single tissue alternative to primary monkey kidney. The combination of MDCK and LLC-MK2 would provide optimal sensitivity for isolation of all six viruses. PMID:227920

Frank, A L; Couch, R B; Griffis, C A; Baxter, B D

1979-01-01

146

Isolation and characterization of viruses from fetal calf serum  

Microsoft Academic Search

Summary  Ten lots of specially procured fetal calf serum collected under sterile conditions and not filtered and 16 lots of commercial\\u000a fetal calf serum were tested for both human and bovine viral contamination. The presence of viruses was evaluated by observing\\u000a for cytopathogenic effect (CPE), hemadsorption with guinea pig erythrocytes, and interference with cytopathogenic challenge\\u000a viruses in both embryonic bovine trachea

C. W. Molander; A. J. Kniazeff; C. W. Boone; A. Paley; D. T. Imagawa

1971-01-01

147

Comparison of monoclonal antibody-based sandwich enzyme-linked immunosorbent assay and virus isolation for detection of peste des petits ruminants virus in goat tissues and secretions.  

PubMed Central

A monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (S-ELISA) was developed for specific detection of peste des petits ruminants virus. Compared with virus isolation in Vero cell cultures using 89 paired tissue and secretion samples from six experimentally infected goats, S-ELISA was significantly more sensitive (71.9% versus 65.2%; P < 0.05). The S-ELISA is a suitable alternative to virus isolation. PMID:8051266

Saliki, J T; House, J A; Mebus, C A; Dubovi, E J

1994-01-01

148

Complete genome sequence analysis of an American isolate of Grapevine virus E.  

PubMed

The complete genome sequence of Grapevine virus E (GVE) collected from a red-berried wine grape cultivar (Cabernet Sauvignon) in Washington State was determined. The 7,568 nucleotide long genome of GVE is more similar in sequence identity with a GVE isolate from a wine grape cv. Shiraz from South Africa when compared with an isolate from "Aki Queen" grape from Japan. Like GVE isolates from South Africa and Japan, the Washington isolate encodes five open reading frames (ORFs) and the overall genome organization is identical among these isolates. In addition to AlkB domain, a DExD domain, belonging to the DEAD-like helicases superfamily, was present upstream of the helicase domain in the replicase ORF of the virus. PMID:23296875

Alabi, Olufemi J; Poojari, Sudarsana; Sarver, Kara; Martin, Robert R; Naidu, Rayapati A

2013-06-01

149

Avian paramyxoviruses and influenza viruses isolated from mallard ducks ( Anas platyrhynchos ) in New Zealand  

Microsoft Academic Search

Summary.  ?A comprehensive study using virological and serological approaches was carried out to determine the status of live healthy\\u000a mallard ducks (Anas platyrhynchos) in New Zealand for infections with avian paramyxoviruses (APMV) and influenza viruses (AIV). Thirty-three viruses isolated\\u000a from 321 tracheal and cloacal swabs were characterized as: 6 AIV (two H5N2 and four H4N6), 10 APMV-1 and 17 APMV-4. Of

W. L. Stanislawek; C. R. Wilks; J. Meers; G. W. Horner; D. J. Alexander; R. J. Manvell; J. A. Kattenbelt; A. R. Gould

2002-01-01

150

Phylogeography of rabies virus isolated from dogs in Brazil between 1985 and 2006  

Microsoft Academic Search

To establish the phylogeographic relationships in rabies viruses in Brazil, we studied a dataset retrieved from GenBank consisting\\u000a of 71 genetic sequences from the coding region of the N gene of rabies viruses isolated in dogs over a period of 22 years.\\u000a The Bayesian Markov chain Monte Carlo method available in the BEAST package was used with the GTR+G+?4 evolutionary model

Pedro Carnieli Jr; Rafael de Novaes Oliveira; Carla Isabel Macedo; Juliana Galera Castilho

2011-01-01

151

Avian flu: Isolation of drug-resistant H5N1 virus  

Microsoft Academic Search

The persistence of H5N1 avian influenza viruses in many Asian countries and their ability to cause fatal infections in humans have raised serious concerns about a global flu pandemic. Here we report the isolation of an H5N1 virus from a Vietnamese girl that is resistant to the drug oseltamivir, which is an inhibitor of the viral enzyme neuraminidase and is

Q. Mai Le; Maki Kiso; Kazuhiko Someya; Yuko T. Sakai; T. Hien Nguyen; Khan H. L. Nguyen; N. Dinh Pham; Ha H. Ngyen; Shinya Yamada; Yukiko Muramoto; Taisuke Horimoto; Ayato Takada; Hideo Goto; Takashi Suzuki; Yasuo Suzuki; Yoshihiro Kawaoka

2005-01-01

152

[Genetic characterization analysis on epidemic rubella virus strains isolated in Liaoning from 2007 to 2012].  

PubMed

To analyze the genetic characterization of epidemic rubella virus strains isolated in Liaoning from 2007-2012, a total of 145 rubella virus strains were isolated using Vero/Slam cell line from the patients' throat swabs during rubella outbreaks and sporadics cases in Liaoning Province from 2007 to 2012. Fragments of 945 nucleotides containing 1E gene from 145 rubella virus isolates were amplified by RT-PCR, the PCR products were sequenced and analyzed. Based on the 739 nucleotides of 1E gene, the phylogenetic trees were constructed with 32 WHO rubella reference strains of 13 genotypes downloaded from GenBank and 145 rubella virus strains. The results showed that the 145 rubella virus strains in 2007 -2012 belonged to genotype 1E, nucleotide acids and amino acids similarities were 97.2%-100.0% and 97.6%-100.0%, respectively. Compared to the 1E reference strains(Rvi/ Dezhou.CHN/02, RVi/MYS/01), the nucleotide acids and amino acids similarities were 96.6%-99.2% and 98.2%-100.0%, respectively except for one amino acid change (Val246-Ala246) of RVi/Shenyang. Liaoning. CHN/13.11/13, and Asp262-Asn262 of RVi/Shenyang. Liaoning. CHN/13.11/4 and RVi/Liaoyang. Liaoning. CHN/26. 11/2. there had no change found in the important antigenic epitope sites, the hemagglutination inhibition and neutralization epitopes of the other rubella viruses. All the 145 strains isolated had the same amino acid change (Leu338--Phe338) in E1 protein. These findings suggested that genotype 1E of rubella virus was the predominant genotype in Liaoning province. the rubella prevailed in recent six years was mainly caused by rubella viruses genotype 1E with multi-transmission routes. PMID:24520763

Wang, Yan; Ma, Yan; Xu, Xiao-Ting; Fan, Xue-Song; Lin, Qian; Sui, Dan; Yin, Ye; Wu, Feng-Tong; Pan, Bai-Ling; Liu, Guang-Yuan; Wang, Ji-Jian; Han, Yue; Guo, Jun-Qiao; Zhao, Zhuo

2013-11-01

153

Analysis of new aphid lethal paralysis virus (ALPV) isolates suggests evolution of two ALPV species.  

PubMed

Aphid lethal paralysis virus (ALPV; family Dicistroviridae) was first isolated from the bird cherry-oat aphid, Rhopalosiphum padi. ALPV-like virus sequences have been reported from many insects and insect predators. We identified a new isolate of ALPV (ALPV-AP) from the pea aphid, Acyrthosiphon pisum, and a new isolate (ALPV-DvV) from western corn rootworm, Diabrotica virgifera virgifera. ALPV-AP has an ssRNA genome of 9940 nt. Based on phylogenetic analysis, ALPV-AP was closely related to ALPV-AM, an ALPV isolate from honeybees, Apis mellifera, in Spain and Brookings, SD, USA. The distinct evolutionary branches suggested the existence of two lineages of the ALPV virus. One consisted of ALPV-AP and ALPV-AM, whilst all other isolates of ALPV grouped into the other lineage. The similarity of ALPV-AP and ALPV-AM was up to 88?% at the RNA level, compared with 78-79?% between ALPV-AP and other ALPV isolates. The sequence identity of proteins between ALPV-AP and ALPV-AM was 98-99?% for both ORF1 and ORF2, whilst only 85-87?% for ORF1 and 91-92?% for ORF2 between ALPV-AP and other ALPV isolates. Sequencing of RACE (rapid amplification of cDNA ends) products and cDNA clones of the virus genome revealed sequence variation in the 5' UTRs and in ORF1, indicating that ALPV may be under strong selection pressure, which could have important biological implications for ALPV host range and infectivity. Our results indicated that ALPV-like viruses infect insects in the order Coleoptera, in addition to the orders Hemiptera and Hymenoptera, and we propose that ALPV isolates be classified as two separate viral species. PMID:25170050

Liu, Sijun; Vijayendran, Diveena; Carrillo-Tripp, Jimena; Miller, W Allen; Bonning, Bryony C

2014-12-01

154

Phenotypic characteristics of human immunodeficiency virus type 1 subtype C isolates of Ethiopian AIDS patients.  

PubMed

It has been estimated that, to date, about 48% of all HIV-infected people in the world carry HIV-1 subtype C virus. Therefore, it is of great importance to gain better knowledge about the genetic and biological characteristics of this virus subtype. In the present study, the biological properties of HIV-1 isolates obtained from nine Ethiopian patients with AIDS were studied. DNA sequencing of the V3 loop of gp120 classified the isolates as subtype C. In primary isolation cultures, virus infection was accompanied by syncytium formation and cell lysis. Interestingly, when examining the growth in primary monocyte-macrophage cultures, initial low-level virus replication was followed by a nonproductive state, from which virus could be rescued by cocultivation with Jurkat(tat) cells. Furthermore, none of the isolates replicated in T cell lines (CEM, MT-2, HuT-78, and H9) or in the promonocytic cell line U937 clone 2. All isolates could use CCR5 as coreceptor, whereas no isolates could use CCR2b, CCR3, CCR5, CXCR4, Bonzo/STRL33, or BOB/GPR15. The genotype of the V3 region correlated with the MT-2 negative/non-syncytium-inducing (NSI) phenotype. Comparative studies revealed that the scarcity of CXCR4 usage as well as other phenotypic characteristics of subtype C isolates distinguish this subtype. On the basis of these data, we suggest that in addition, factors other than viral phenotype may govern the pathogenic potential of subtype C isolates. PMID:10331443

Björndal, A; Sönnerborg, A; Tscherning, C; Albert, J; Fenyö, E M

1999-05-01

155

Isolation of influenza virus in human lung embryonated fibroblast cells (MRC-5) from clinical samples.  

PubMed

Ninety-four pharyngeal swab samples corresponding to 94 patients with suspected influenza virus infection were inoculated in Madin-Darby canine kidney (MDCK) cells, the conventional cell system for the isolation of influenza virus, and in fibroblastic human embryo lung (MRC-5) cells, a cell system less commonly used for this purpose but one frequently used in clinical virology laboratories. Both cell preparations were treated with trypsin. Influenza virus was recovered from 15% of the samples inoculated in MDCK cells and from 18% of those inoculated in MRC-5 cells. The use of MRC-5 cells can simplify the search for respiratory viruses and would assist in the rapid detection of influenza virus during new epidemics. PMID:7665680

de Ońa, M; Melón, S; de la Iglesia, P; Hidalgo, F; Verdugo, A F

1995-07-01

156

Acanthamoeba polyphaga mimivirus Stability in Environmental and Clinical Substrates: Implications for Virus Detection and Isolation  

PubMed Central

Viruses are extremely diverse and abundant and are present in countless environments. Giant viruses of the Megavirales order have emerged as a fascinating research topic for virologists around the world. As evidence of their ubiquity and ecological impact, mimiviruses have been found in multiple environmental samples. However, isolation of these viruses from environmental samples is inefficient, mainly due to methodological limitations and lack of information regarding the interactions between viruses and substrates. In this work, we demonstrate the long-lasting stability of mimivirus in environmental (freshwater and saline water) and hospital (ventilator plastic device tube) substrates, showing the detection of infectious particles after more than 9 months. In addition, an enrichment protocol was implemented that remarkably increased mimivirus detection from all tested substrates, including field tests. Moreover, biological, morphological and genetic tests revealed that the enrichment protocol maintained mimivirus particle integrity. In conclusion, our work demonstrated the stability of APMV in samples of environmental and health interest and proposed a reliable and easy protocol to improve giant virus isolation. The data presented here can guide future giant virus detection and isolation studies. PMID:24498379

de Almeida, Gabriel M.; Campos, Rafael K.; Boratto, Paulo V. M.; Franco-Luiz, Ana P. M.; La Scola, Bernard; Ferreira, Paulo C. P.; Kroon, Erna G.; Abrahăo, Jônatas S.

2014-01-01

157

Isolation of a protein kinase induced by herpes simplex virus type 1  

SciTech Connect

Researchers have isolated a new cyclic AMP-independent protein kinase activity induced in HeLa cells by infection with herpes simplex virus type 1. Induction of the enzyme does not occur in cells treated with cycloheximide at the time of infection, or in cells infected with UV-inactivated herpes simplex virus type 1. The amount of enzyme induced in infected cells is dependent upon the multiplicity of infection. An enzyme with identical properties to the appearing in infected HeLa cells is also induced by herpes simplex virus type 1 in BHK cells.

Blue, W.T.; Stobbs, D.G.

1981-04-01

158

Genomic and phylogenetic characterization of Leanyer virus, a novel orthobunyavirus isolated in northern Australia  

PubMed Central

Leanyer virus (LEAV), currently classified as a member of the genus Orthobunyavirus, in the family Bunyaviridae, was originally isolated from a pool of Anopheles meraukensis mosquitoes, collected at Leanyer, Northern Territory, Australia in 1974. When it failed to react in serological tests with antisera from other known viruses, full-length genomic sequencing was pursued to determine the relationship of LEAV to other orthobunyavirus species. Genetic and serological characterization confirmed its antigenic distance from other orthobunyaviruses, including to its closest genetic neighbours, the Simbu group viruses, suggesting that it may represent a new antigenic complex. PMID:21402599

Savji, Nazir; Travassos da Rosa, Amelia; Hutchison, Stephen; Celone, Christopher; Hui, Jeffrey; Briese, Thomas; Calisher, Charles H.; Lipkin, W. Ian

2011-01-01

159

Segment 10 based molecular epidemiology of bluetongue virus (BTV) isolates from Turkey: 1999-2001.  

PubMed

Bluetongue is a significant arbovirus infection that has a negative impact on ruminant productivity in Turkey. Twenty-one Turkish BTV isolates were analyzed phylogenetically, based on genome segment 10 (Seg-10) nucleotide sequences. These analyses were used to explore the epidemiological background of individual isolates from both a regional and global perspective. In the regional analysis, the different BTV strains fell into two groups (Group 1 and Group 2). The Turkish virus isolates were localized in Group 1 which contains two sub-groups. The neighbor-joining analysis revealed that Seg-10 of majority of the Turkish viruses was closely related to certain other virus strains allocated in the eastern lineage. The Seg-10's of two viruses (TR25 and TR26) were more closely related to strains isolated in the Asia-Australia region. These strains belong to the 'eastern' topotype identified by [Maan, S., Maan, N.S., Ross-Smith, N., Batten, C.A., Shaw, A.E., Anthony, S.J., Samuel, A.R., Darpel, K.E., Veronesi, E., Oura, C.A.L., Singh,K.P., Nomikou, K., Potgieter, A.C., Attoui, H., van Rooij, E., van Rijn, P., De Clercq, K., Vandenbussche, F., Zientara, S., Bréard, E., Sailleau, C., Beer, M., Hoffman, B., Mellor, P.S., Mertens, P.P.C., 2008. Sequence analysis of bluetongue virus serotype 8 from the Netherlands 2006 and comparison to other European strains. Virology 377, 308-318]. Comparisons of amino acid sequences deduced from the Seg-10 genes showed a high level of conservation in the NS3/3A proteins from the Turkish viruses. The more frequent amino acid substitutions were identified by multiple alignment analysis, and one of the isolates (TR23) was remarkably found to be genetically quite distinct from the other isolates. PMID:19428746

Ozkul, Aykut; Erturk, Arife; Caliskan, Elvin; Sarac, Fahriye; Ceylan, Cagatay; Mertens, Peter; Kabakli, Ozden; Dincer, Ender; Cizmeci, Sirin G

2009-06-01

160

Thermostability of Subpopulations of H2N3 Influenza Virus Isolates from Mallard Ducks ?  

PubMed Central

Maintenance of avian influenza virus in waterfowl populations requires that virions remain infectious while in the environment. Temperature has been shown to negatively correlate with persistence time, which is the duration for which virions are infectious. However, thermostability can vary between isolates regardless of subtype, and it is not known whether this variation occurs when host and geographic location of isolation are controlled. In this study, we analyzed the thermostabilities of 7 H2N3 viruses isolated from mallard ducks in Alberta, Canada. Virus samples were incubated at 37°C and 55°C, and infectivity titers were calculated at different time points. Based on the rate of infectivity inactivation at 37°C, isolates could be grouped into either a thermosensitive or thermostable fraction for both egg- and MDCK-grown virus populations. Titers decreased more rapidly for isolates incubated at 55°C, and this loss of infectivity occurred in a nonlinear, 2-step process, which is in contrast with the consensus on thermostability. This suggests that stock samples contain a mixture of subpopulations with different thermostabilities. The rate of decrease for the sensitive fraction was approximately 14 times higher than that for the stable fraction. The presence of subpopulations is further supported by selection experiments and plaque purification, both of which result in homogenous populations that exhibit linear decreases of infectivity titer. Therefore, variation of thermostability of influenza virus isolates begins at the level of the population. The presence of subpopulations with high thermostability suggests that avian viruses can persist in water longer than previously estimated, thus increasing the probability of transmission to susceptible hosts. PMID:20610728

Negovetich, Nicholas J.; Webster, Robert G.

2010-01-01

161

Evolutionary Changes Affecting Rapid Identification of 2008 Newcastle Disease Viruses Isolated from Double-Crested Cormorants? †  

PubMed Central

A morbidity-mortality event involving virulent Newcastle disease virus (NDV) in wild double-crested cormorants (Phalacrocorax auritus) occurred in North America in the summer of 2008. All 22 viruses isolated from cormorants were positively identified by the USDA-validated real-time reverse transcription-PCR assay targeting the matrix gene. However, the USDA-validated reverse transcription-PCR assay targeting the fusion gene that is specific for virulent isolates identified only 1 of these 22 isolates. Additionally, several of these isolates have been sequenced, and this information was used to identify genomic changes that caused the failure of the test and to revisit the evolution of NDV in cormorants. The forward primer and fusion probe were redesigned from the 2008 cormorant isolate sequence, and the revised fusion gene test successfully identified all 22 isolates. Phylogenetic analyses using both the full fusion sequence and the partial 374-nucleotide sequence identified these isolates as genotype V, with their nearest ancestor being an earlier isolate collected from Nevada in 2005. Histopathological analysis of this ancestral strain revealed morphological changes in the brain consistent with that of the traditional mesogenic pathotypes in cormorants. Intracerebral pathogenicity assays indicated that each of these isolates is virulent with values of >0.7 but not more virulent than earlier isolates reported from Canada. PMID:20107098

Rue, Cary A.; Susta, Leonardo; Brown, Corrie C.; Pasick, John M.; Swafford, Seth R.; Wolf, Paul C.; Killian, Mary Lea; Pedersen, Janice C.; Miller, Patti J.; Afonso, Claudio L.

2010-01-01

162

Evolutionary changes affecting rapid identification of 2008 Newcastle disease viruses isolated from double-crested cormorants.  

PubMed

A morbidity-mortality event involving virulent Newcastle disease virus (NDV) in wild double-crested cormorants (Phalacrocorax auritus) occurred in North America in the summer of 2008. All 22 viruses isolated from cormorants were positively identified by the USDA-validated real-time reverse transcription-PCR assay targeting the matrix gene. However, the USDA-validated reverse transcription-PCR assay targeting the fusion gene that is specific for virulent isolates identified only 1 of these 22 isolates. Additionally, several of these isolates have been sequenced, and this information was used to identify genomic changes that caused the failure of the test and to revisit the evolution of NDV in cormorants. The forward primer and fusion probe were redesigned from the 2008 cormorant isolate sequence, and the revised fusion gene test successfully identified all 22 isolates. Phylogenetic analyses using both the full fusion sequence and the partial 374-nucleotide sequence identified these isolates as genotype V, with their nearest ancestor being an earlier isolate collected from Nevada in 2005. Histopathological analysis of this ancestral strain revealed morphological changes in the brain consistent with that of the traditional mesogenic pathotypes in cormorants. Intracerebral pathogenicity assays indicated that each of these isolates is virulent with values of >0.7 but not more virulent than earlier isolates reported from Canada. PMID:20107098

Rue, Cary A; Susta, Leonardo; Brown, Corrie C; Pasick, John M; Swafford, Seth R; Wolf, Paul C; Killian, Mary Lea; Pedersen, Janice C; Miller, Patti J; Afonso, Claudio L

2010-07-01

163

Effectiveness of reverse transcription-PCR, virus isolation, and enzyme-linked immunosorbent assay for diagnosis of influenza A virus infection in different age groups.  

PubMed

The degrees of effectiveness of reverse transcription (RT)-PCR, virus isolation, and antigen enzyme-linked immunosorbent assay (ELISA) for the detection of influenza A virus were evaluated with nasopharyngeal swabs from 150 patients (1 week to 86 years old) with influenza A virus infection. RT-PCR had a sensitivity for influenza A virus in stock virus preparations 10(3) times higher than virus isolation and 10(6) to 10(7) times higher than ELISA. The detection rate achieved by RT-PCR in clinical samples was clearly higher (93%) than that by virus isolation (80%) and ELISA (62%). Despite low overall detection rates achieved by antigen ELISA, samples from patients younger than 5 years old yielded higher-than-average rates in this rapid assay (88%). The likelihood of negative results in the ELISA increased significantly with increasing age of the patient (P < 0.01). The degrees of effectiveness of RT-PCR and virus isolation were not influenced by the age of the patient. Neither influenza immunizations nor the interval between onset of symptoms and laboratory investigation (mean, 4.7 days; standard deviation, 3.3 days) affected results obtained by the three test systems. Our results demonstrate that the ELISA is reliable for rapid laboratory diagnosis of influenza in infants and young children, but for older patients application of RT-PCR or virus isolation is necessary to avoid false negative results. PMID:12037063

Steininger, Christoph; Kundi, Michael; Aberle, Stephan W; Aberle, Judith H; Popow-Kraupp, Theresia

2002-06-01

164

Genetic characterization of H5N1 influenza A viruses isolated from zoo tigers in Thailand  

Microsoft Academic Search

The H5N1 avian influenza virus outbreak among zoo tigers in mid-October 2004, with 45 animals dead, indicated that the avian influenza virus could cause lethal infection in a large mammalian species apart from humans. In this outbreak investigation, six H5N1 isolates were identified and two isolates (A\\/Tiger\\/Thailand\\/CU-T3\\/04 and A\\/Tiger\\/Thailand\\/CU-T7\\/04) were selected for whole genome analysis. Phylogenetic analysis of the 8

Alongkorn Amonsin; Sunchai Payungporn; Apiradee Theamboonlers; Roongroje Thanawongnuwech; Sanipa Suradhat; Nuananong Pariyothorn; Rachod Tantilertcharoen; Sudarat Damrongwantanapokin; Chantanee Buranathai; Arunee Chaisingh; Thaweesak Songserm; Yong Poovorawan

2006-01-01

165

Characterization of H5N1 influenza A viruses isolated from domestic green-winged teal  

Microsoft Academic Search

Two avian influenza virus strains, A\\/domestic green-winged teal\\/Hunan\\/67\\/2005 (H5N1) (D-GWT\\/67) and A\\/domestic green-winged\\u000a teal\\/Hunan\\/79\\/2005 (H5N1) (D-GWT\\/79), were isolated from healthy domestic green-winged teals (Anas crecca) in Hunan Province, South China. Genomic analysis showed that both isolates were reassortants. The hemagglutinin (HA) genes\\u000a of the two isolates were closely related to that of an H5N1 strain isolated from tree sparrow (A\\/tree

Jianjun Chen; Zhongdong Yang; Quanjiao Chen; Xueying Liu; Fang Fang; Haiyan Chang; Dongmei Li; Ze Chen

2009-01-01

166

Isolation of Borna Disease Virus from Human Brain Tissue  

Microsoft Academic Search

Serological and molecular epidemiological studies indicate that Borna disease virus (BDV) can infect humans and is possibly associated with certain neuropsychiatric disorders. We examined brain tissue collected at autopsy from four schizophrenic patients and two healthy controls for the presence of BDV markers in 12 different brain regions. BDV RNA and antigen was detected in four brain regions of a

YURIE NAKAMURA; HIROKAZU TAKAHASHI; YUKO SHOYA; TAKAAKI NAKAYA; MAKIKO WATANABE; KEIZO TOMONAGA; KAZUHIKO IWAHASHI; KIYOSHI AMENO; NORIKO MOMIYAMA; HIROYUKA TANIYAMA; TETSUTARO SATA; TAKESHI KURATA; JUAN CARLOS DE LA TORRE; KAZUYOSHI IKUTA

2000-01-01

167

Infectious hematopoietic necrosis virus: Monophyletic origin of European isolates from North American Genogroup M  

USGS Publications Warehouse

Infectious hematopoietic necrosis virus (IHNV) was first detected in Europe in 1987 in France and Italy, and later, in 1992, in Germany. The source of the virus and the route of introduction are unknown. The present study investigates the molecular epidemiology of IHNV outbreaks in Germany since its first introduction. The complete nucleotide sequences of the glycoprotein (G) and non-virion (NV) genes from 9 IHNV isolates from Germany have been determined, and this has allowed the identification of characteristic differences between these isolates. Phylogenetic analysis of partial G gene sequences (mid-G, 303 nucleotides) from North American IHNV isolates (Kurath et al. 2003) has revealed 3 major genogroups, designated U, M and L. Using this gene region with 2 different North American IHNV data sets, it was possible to group the European IHNV strains within the M genogroup, but not in any previously defined subgroup. Analysis of the full length G gene sequences indicated that an independent evolution of IHN viruses had occurred in Europe. IHN viruses in Europe seem to be of a monophyletic origin, again most closely related to North American isolates in the M genogroup. Analysis of the NV gene sequences also showed the European isolates to be monophyletic, but resolution of the 3 genogroups was poor with this gene region. As a result of comparative sequence analyses, several different genotypes have been identified circulating in Europe. ?? Inter-Research 2005.

Enzmann, P.-J.; Kurath, G.; Fichtner, D.; Bergmann, S.M.

2005-01-01

168

Characterization of Newcastle disease virus isolated from northern pintail (Anas acuta) in Japan.  

PubMed

A field isolate of Newcastle disease virus (NDV) isolated from northern pintail (Anas acuta) in Tohoku district, northeast Japan, was characterized. Phylogenetic analysis of the fusion protein indicated that the isolate belonged to genotype I and was closely related to isolates from the Far East corresponded to the migration route for this bird species. The isolate had the typical avirulent cleavage site of the fusion protein (112)GKQGR*L(117). In addition, pathogenicity tests indicated the isolate to have avirulent characteristics. However, the isolate has been shown to cause fusion cytopathic effects and form plaques on chicken embryo fibroblasts (CEF) in the absence of trypsin. The present results suggest that the CEF-adapted NDV, which is avirulent, is circulating among waterfowl populations. PMID:18176032

Sakai, Kouji; Sakabe, Genki; Tani, Orie; Watanabe, Yuko; Jahangir, Alam; Nakamura, Masayuki; Takehara, Kazuaki

2007-12-01

169

A unique isolate of beak and feather disease virus isolated from budgerigars (Melopsittacus undulatus) in South Africa.  

PubMed

Beak and feather disease virus (BFDV), the causative agent of psittacine beak and feather disease (PBFD) infects psittaciformes worldwide. We provide an annotated sequence record of three full-length unique genomes of BFDV isolates from budgerigars (Melopsittacus undulatus) from a breeding farm in South Africa. The isolates share >99% nucleotide sequence identity with each other and approximately 96% nucleotide sequence identity to two recent isolates (Melopsittacus undulatus) from Thailand but only between 91.6 and 86.6% identity with all other full-length BFDV sequences. Maximum-likelihood analysis and recombination analysis suggest that the South African budgerigar BFDV isolates are unique to budgerigars, are non-recombinant in origin, and represent a new genotype of BFDV. PMID:20127375

Varsani, Arvind; de Villiers, Gillian K; Regnard, Guy L; Bragg, Robert R; Kondiah, Kulsum; Hitzeroth, Inga I; Rybicki, Edward P

2010-03-01

170

Receptor-Binding and Oncogenic Properties of Polyoma Viruses Isolated from Feral Mice  

PubMed Central

Laboratory strains of the mouse polyoma virus differ markedly in their abilities to replicate and induce tumors in newborn mice. Major determinants of pathogenicity lie in the sialic binding pocket of the major capsid protein Vp1 and dictate receptor-binding properties of the virus. Substitutions at two sites in Vp1 define three prototype strains, which vary greatly in pathogenicity. These strains replicate in a limited fashion and induce few or no tumors, cause a disseminated infection leading to the development of multiple solid tumors, or replicate and spread acutely causing early death. This investigation was undertaken to determine the Vp1 type(s) of new virus isolates from naturally infected mice. Compared with laboratory strains, truly wild-type viruses are constrained with respect to their selectivity and avidity of binding to cell receptors. Fifteen of 15 new isolates carried the Vp1 type identical to that of highly tumorigenic laboratory strains. Upon injection into newborn laboratory mice, the new isolates induced a broad spectrum of tumors, including ones of epithelial as well as mesenchymal origin. Though invariant in their Vp1 coding sequences, these isolates showed considerable variation in their regulatory sequences. The common Vp1 type has two essential features: 1) failure to recognize “pseudoreceptors” with branched chain sialic acids binding to which would attenuate virus spread, and 2) maintenance of a hydrophobic contact with true receptors bearing a single sialic acid, which retards virus spread and avoids acute and potentially lethal infection of the host. Conservation of these receptor-binding properties under natural selection preserves the oncogenic potential of the virus. These findings emphasize the importance of immune protection of neonates under conditions of natural transmission. PMID:18085820

Velupillai, Palanivel; Dahl, Jean; Telford, Samuel; Bronson, Roderick; Benjamin, Thomas

2007-01-01

171

Isolation and Phylogenetic Analysis of Novel Viruses Infecting the Phytoplankton Phaeocystis globosa (Prymnesiophyceae)  

PubMed Central

Viruses infecting the harmful bloom-causing alga Phaeocystis globosa (Prymnesiophyceae) were readily isolated from Dutch coastal waters (southern North Sea) in 2000 and 2001. Our data show a large increase in the abundance of putative P. globosa viruses during blooms of P. globosa, suggesting that viruses are an important source of mortality for this alga. In order to examine genetic relatedness among viruses infecting P. globosa and other phytoplankton, DNA polymerase gene (pol) fragments were amplified and the inferred amino acid sequences were phylogenetically analyzed. The results demonstrated that viruses infecting P. globosa formed a closely related monophyletic group within the family Phycodnaviridae, with at least 96.9% similarity to each other. The sequences grouped most closely with others from viruses that infect the prymnesiophyte algae Chrysochromulina brevifilum and Chrysochromulina strobilus. Whether the P. globosa viruses belong to the genus Prymnesiovirus or form a separate group needs further study. Our data suggest that, like their phytoplankton hosts, the Chrysochromulina and Phaeocystis viruses share a common ancestor and that these prymnesioviruses and their algal host have coevolved. PMID:15184176

Brussaard, C. P. D.; Short, S. M.; Frederickson, C. M.; Suttle, C. A.

2004-01-01

172

Isolation of virus from brain after immunosuppression of mice with latent herpes simplex  

NASA Astrophysics Data System (ADS)

Herpes simplex virus (HSV) is usually present in a latent form in the trigeminal ganglion of man1-3. Various stress factors may induce virus reactivation, which is manifest by a lip lesion (innervated from the trigeminal ganglion) and the production of infectious virus. The considerable experimental efforts to define the conditions that lead to the reactivation of latent HSV have concentrated on isolating virus either from the original extraneural site of virus inoculation, or from cell-free homogenates of sensory ganglia from latently infected animals4-15. Recent DNA hybridization experiments resulted in the demonstration of the presence of HSV genomes in the brain tissue of both latently infected mice, and of humans who showed no clinical symptoms of HSV (ref. 16 and N. Fraser, personal communication). This led us to consider the possibility that HSV may be present in brain tissue as the result of either reactivation of the virus in brain cells or the passage of reactivated virus from trigeminal ganglia through the brain stem to the brain. The presence of infectious HSV in brain tissue has not previously been demonstrated; yet this could be a factor in chronic, relapsing neurological diseases such as multiple sclerosis. We have now shown experimentally that mice carrying latent HSV in their trigeminal ganglia may, following massive immunosuppression, express infectious virus in the central nervous system (CNS).

Kastrukoff, Lorne; Long, Carol; Doherty, Peter C.; Wroblewska, Zofia; Koprowski, Hilary

1981-06-01

173

Two Viruses Isolated from Rodents (Clethrionomys gapperi and Microtus pennsylvanicus) Trapped in St. Lawrence County, New York  

Microsoft Academic Search

Four strains of C. gapperi virus were isolated from 3 Clethrionomys gapperi and 47 strains of Microtus virus from 15 Microtus pennsylvani- cus and 1 Mus musculus. One of the Microtus strains was isolated from a poo1 of 20 mites while the others were from rodent tissues. These agents were insensitive to ether and sodium desoxycholate, withstood freezing at -70

ELINOR WHITNEY; ALBERT P. ROZ; GEORGE A. RAYNER

174

Phylogenetic relationships of the G gene sequence of bovine ephemeral fever virus isolated in Japan, Taiwan and Australia  

Microsoft Academic Search

The G gene encoding the neutralization antigen of bovine ephemeral fever virus (BEFV) was characterized in order to define the virus's molecular epidemiology in Japan and the genetic relationships among the Japanese, Taiwanese and Australian isolates. The nucleotide and amino acid sequences of the gene were highly conserved among the Japanese strains, regardless of the year of isolation, and were

Tomoko Kato; Maki Aizawa; Katsunori Takayoshi; Tamotsu Kokuba; Tohru Yanase; Hiroaki Shirafuji; Tomoyuki Tsuda; Makoto Yamakawa

2009-01-01

175

Recent human influenza A (H1N1) viruses are closely related genetically to strains isolated in 1950  

Microsoft Academic Search

Comparison of the oligonucleotide maps of the RNAs of current human influenza (H1N1) virus isolates shows these strains to be much more closely related to viruses isolated in 1950 than to strains which circulated before or after that period.

Katsuhisa Nakajima; Ulrich Desselberger; Peter Palese

1978-01-01

176

Genetic Characterization and Geographic Distribution of Rabies Virus Isolates in Brazil: Identification of Two Reservoirs, Dogs and Vampire Bats  

Microsoft Academic Search

We analyzed 50 rabies virus samples isolated in Brazil from 12 dogs, 11 cats, 5 vampire bats, 15 cattle, 2 horses, 1 pig, 1 sheep, and 3 humans to investigate the molecular epidemiology of rabies viruses. We sequenced 203 nucleotides on the nucleoprotein gene by direct sequencing of the PCR-amplified products. All the isolates belonged to the genotype 1 and

Mikako Ito; Yohko T. Arai; Takuya Itou; Takeo Sakai; Fumio H. Ito; Tomohiko Takasaki; Ichiro Kurane

2001-01-01

177

Full genome sequence and some biological properties of reticuloendotheliosis virus strain APC566 isolated from endangered Attwater's prairie chickens  

Microsoft Academic Search

Reticuloendotheliosis virus (REV) causes runting, high mortality, immunosuppression, and chronic neoplasia associated with T and\\/or B cell lymphomas in a variety of domestic and wild birds, including Attwater's prairie chickens (APC) (Tympanuchus cupido attwateri). The complete proviral sequence of a recent REV isolate from APC (REV APC-566) was determined. This virus was isolated from an APC maintained in captivity in

Taylor Barbosa; Guillermo Zavala; Sunny Cheng; Pedro Villegas

2007-01-01

178

Biological and molecular variation of Iranian Cauliflower mosaic virus (CaMV) isolates.  

PubMed

Seventeen provinces of Iran were surveyed during 2003-2012 to find Brassicaceae hosts of Cauliflower mosaic virus (CaMV). A total 397 samples were collected from plants with virus-like symptoms. Among those tested by ELISA, 255 samples (67.2 %) were found to be infected with CaMV. Mechanical transmission tests showed that the Iranian isolates have similar biological properties on a number of Brassica and Raphanus plant species and cultivars tested. However, the isolates varied in the severity of symptoms they induced and in the capacity to infect B. oleracea var. capitata, on the basis of which they were grouped into two distinct biotypes L/MMo (latent/mild mottle) and severe (S) infection. The molecular diversity of natural population of CaMV were investigated based on the complete sequences of OFR 6 of 36 Iranian isolates collected from different geographically distant regions in Iran alongside the sequences of 14 previously reported isolates. Phylogenetic analyses indicated that the Iranian CaMV isolates belong to two groups (GI and GII). Most of the Iranian isolates fell into GI with other exotic isolates; however, the isolates from North-East Iran with Xinjiang from China fell into GII. The phylogenetic group GII (the North-East Iranian isolates) closely corresponded to the S biological group however other Iranian isolates corresponded to the L/MMo biological group. The within-population diversity was lower than the between population diversity suggesting the contribution of a founder effect on diversification of CaMV isolates. The Iranian isolates were differentiated from other exotic CaMV isolates and clustered into two RFLP groups using Hpy99I which closely corresponded to the biological and phylogenetic groups. This study showed the evolutionary process in CaMV isolates is shaped by a combination of host range differentiation and nucleotide substitution using the approach of population genetics. PMID:23828619

Farzadfar, Shirin; Pourrahim, Reza

2013-10-01

179

Isolation and characterization of bovine parainfluenza virus type 3 from water buffaloes (Bubalus bubalis) in Argentina  

PubMed Central

Background Parainfluenza virus type 3 (PIV3) was isolated from dairy buffaloes (Bubalus bubalis) naturally affected with respiratory and reproductive clinical conditions. Results Examination of nasal and vaginal swabs collected from 12 diseased buffaloes led to the isolation of three paramyxovirus isolates from two animals. Antigenic, morphological and biological characteristics of these three isolates were essentially similar to those of members of the Paramyxoviridae family. Antigenic analysis by direct immunofluorescence and cross neutralization test placed these isolates together with bovine parainfluenza virus type 3 (BPIV3). Nucleotide and amino acid phylogenetic analysis of partial matrix gene sequences of the buffalo isolates and six field BPIV3 isolates from bovines in Argentina were studied. Buffalo isolates were similar to genotype B (BPIV3b) while the six BPIV3 isolates were similar to genotypes A (BPIV3a) and C (BPIV3c). Conclusions This is the first characterization of BPIV3 in water buffalo. According to the samples analyzed, in Argentina, the genotype B was found in buffalo and the genotypes A and C were found in cattle. PMID:22716217

2012-01-01

180

Comparison of the coat protein genes of Mirafiori lettuce big-vein virus isolates from Australia with those of isolates from other continents  

Microsoft Academic Search

The complete coat protein nucleotide encoding sequences of 13 Mirafiori lettuce big-vein virus isolates from Australia were compared to those of 23 other isolates, including one from Australia. On phylogenetic analysis,\\u000a sub-clade A1 contained isolates from Australia (13), Europe and Japan, A2 contained isolates from Australia (1), Europe and\\u000a South America, and B1 and B2 contained only European isolates. In

Linda D. Maccarone; Martin J. Barbetti; Krishnapillai Sivasithamparam; Roger A. C. Jones

2010-01-01

181

Isolation of Newcastle disease virus from teals (Anas crecca) in Iran.  

PubMed

Eight of 30 teals (Anas crecca) died several days following capture and Newcastle Disease Virus (NDV) was isolated from all eight. Brains from the dead birds were homogenized and inoculated into chicken embryos. The allantoic fluid from the embryos were inoculated into 10 domestic chickens susceptible to NDV and 10 chickens immunized against NDV. Eight of 10 (80%) susceptible chickens died, while the immunized chickens remained healthy. Anti-NDV serum showed complete homology against NDV and the eight isolates. PMID:480525

Bozorgmehri-Fard, M H; Keyvanfar, H

1979-04-01

182

“Zaliv Terpeniya” virus, a new Uukuniemi group arbovirus isolated from ixodes (Ceratixodes) putus Pick.-Camb. 1878 on tyuleniy island (Sakhalin region) and Commodore islands (Kamchatsk region)  

Microsoft Academic Search

Summary Three virus strains isolated fromIxodes putus ticks were shown to be related to, though not identical, with Uukuniemi virus by complement-fixation tests. No antigenic relations were detected in the cross-neutralization test performed with the virus isolated and Uukuniemi virus. Two virus strains were isolated in 1969 on Tyuleniy island, Zaliv Terpeniya (Patience Bay) of the Sea of Okhotsk (Sakhalin

D. K. Lvov; A. A. TIMOPI-IEEVA; V. L. GROMASttEVSKI; G. V. GOSTINSIICIIIKOVA; O. V. Veselovskaya; V. I. Chervonski; K. B. Fomina; A. I. Gromov; A. G. Pogrebenko; V. Yu. Zhezmer

1973-01-01

183

Coreceptor usage of sequential isolates from cynomolgus monkeys experimentally Infected with simian immunodeficiency virus (SIVsm).  

PubMed

Sequential isolates from eight cynomolgus monkeys experimentally infected with simian immunodeficiency virus (SIVsm, of sooty mangabey origin) were tested for coreceptor use in the human osteosarcoma indicator cell line, GHOST(3), expressing CD4 and one or another of the chemokine receptors CCR3, CCR5, CXCR4, BOB, or the orphan receptor Bonzo. The indicator cell line carries the human immunodeficiency virus type 2 long terminal repeat-driven green fluorescence protein gene that becomes activated upon infection with HIV or SIV and fluorescence can be quantitated by flow cytometric analysis. The methodological details are described in the accompanying paper (Vödrös et al., 2001, Virology 290, in press). All SIVsm inoculum viruses and reisolates used CCR5 with a high level of efficiency. CCR5 use was stable over time. BOB and Bonzo use was less efficient than CCR5 use and, in particular, late isolates obtained at the time of immunodeficiency varied greatly in their coreceptor use and often could not establish a productive infection in BOB- or Bonzo-expressing cells. Unexpectedly, early reisolates obtained 12 days postinfection could infect the entire GHOST(3) panel including the parental cells. In one case this was due to use of CXCR4, either transfected or endogenously expressed on the GHOST(3) cells. Our results demonstrate the complex coreceptor use of SIVsm isolates. Moreover, they focus attention on the initial phase of virus replication when the availability of target cells may govern the replication pattern of the virus. PMID:11878872

Vödrös, D; Thorstensson, R; Biberfeld, G; Schols, D; De Clercq, E; Fenyö, E M

2001-12-01

184

Isolation of mixed subtypes of influenza A virus from a bald eagle (Haliaeetus leucocephalus)  

PubMed Central

From April 2007 to March 2008, cloacal swabs were obtained from 246 casualty raptors recovered by various wildlife rehabilitation centers in the United States. The swabs were placed in a virus transport medium and transported to the laboratory on ice packs. At the laboratory, the samples were pooled with each pool consisting of five samples. All pools (n = 50) were screened for the presence of avian influenza virus (AIV) using a real time reverse transcription-polymerase chain reaction (rRT-PCR); one of the pools was found positive. All five samples in this pool were tested individually by rRT-PCR; one sample from a bald eagle was found positive. This sample was inoculated in embryonated chicken eggs for virus isolation and a hemagglutinating virus was isolated. Complete genome sequencing of the isolate revealed a mixed infection with H1N1 and H2N1 subtypes. Further analysis revealed that the PB1-F2 gene sequence of H1N1 virus had the N66S virulence-associated substitution. Further studies on ecology and epidemiology of AIV in raptors are needed to help understand their role in the maintenance and evolution of AIV. PMID:20667110

2010-01-01

185

Isolation of mixed subtypes of influenza A virus from a bald eagle (Haliaeetus leucocephalus).  

PubMed

From April 2007 to March 2008, cloacal swabs were obtained from 246 casualty raptors recovered by various wildlife rehabilitation centers in the United States. The swabs were placed in a virus transport medium and transported to the laboratory on ice packs. At the laboratory, the samples were pooled with each pool consisting of five samples. All pools (n = 50) were screened for the presence of avian influenza virus (AIV) using a real time reverse transcription-polymerase chain reaction (rRT-PCR); one of the pools was found positive. All five samples in this pool were tested individually by rRT-PCR; one sample from a bald eagle was found positive. This sample was inoculated in embryonated chicken eggs for virus isolation and a hemagglutinating virus was isolated. Complete genome sequencing of the isolate revealed a mixed infection with H1N1 and H2N1 subtypes. Further analysis revealed that the PB1-F2 gene sequence of H1N1 virus had the N66S virulence-associated substitution. Further studies on ecology and epidemiology of AIV in raptors are needed to help understand their role in the maintenance and evolution of AIV. PMID:20667110

Goyal, Sagar M; Jindal, Naresh; Chander, Yogesh; Ramakrishnan, Muthanan A; Redig, Patrick T; Sreevatsan, Srinand

2010-01-01

186

Isolations of Potosi virus from mosquitoes (Diptera: Culicidae) collected in Connecticut.  

PubMed

Potosi virus (POTV) (Bunyaviridae: Orthobunyavirus) was first isolated from Aedes albopictus (Skuse) collected in Potosi, MO, in 1989, and subsequent isolations were reported from Illinois, Michigan, Ohio, and the Carolinas. To determine whether the distribution of this virus extends into the northeastern United States, we analyzed arboviruses acquired from mosquitoes collected in Connecticut from 1998 to 2004. In 2001, a bunyavirus was isolated from Aedes vexans (Meigen) that was different from other arboviruses known to occur in Connecticut by cross-neutralization and reverse transcription-polymerase chain reaction (RT-PCR) assays. Nucleotide and encoded amino acid sequences of a portion of the G2 envelope gene were 99 and 100% similar to POTV, respectively, yet distinct from indigenous strains of Jamestown Canyon (JCV), Cache Valley (CVV), and Trivittatus virus (TVTV). Viral isolates obtained from the statewide surveillance program were retested by RT-PCR coupled with restriction enzyme analysis to distinguish POTV from other bunyaviruses. POTV isolates, previously typed by neutralization, were correctly identified by RT-PCR; however, many isolates classified as JCV or CVV by enzyme-linked immunosorbent assay proved to be POTV by molecular assays. In total, 92 strains of POTV were isolated from 12 mosquito species in 2000, 2001, and 2003, whereas POTV was not detected in mosquitoes sampled during 1998, 1999, 2002, and 2004. Viral isolation rates were highest for Anopheles punctipennis (Say) (3.2-11.3 infection rate per 1,000 mosquitoes), whereas the greatest number of isolates came from Ochlerotatus trivittatus (Coquillett) (8-16 isolates). This finding represents the first detection of POTV in the northeastern United States where it infects a diverse array of mosquito species. PMID:16363172

Armstrong, Philip M; Andreadis, Theodore G; Anderson, John F; Main, Andrew J

2005-09-01

187

Isolation of bovine viral diarrhea virus from a free-ranging mule deer in Wyoming.  

PubMed

A noncytopathic type 1a bovine viral diarrhea virus (BVDV) was isolated from a free-ranging yearling female mule deer (Odocoileus hemionus) from northwestern Wyoming (USA). The mule deer was emaciated, weak, and salivating, and Arcanobacterium pyogenes was cultured from lung abscesses. Bovine viral diarrhea virus was isolated from lung, however, BVDV antigen was not detected by immunohistochemistry. The BVDV genotype was determined by reverse transcriptase polymerase chain reaction and the RNA sequences from the 5'UTR and E2 genes compared with sequences of a type 1a BVDV isolated from cattle from the same area as the deer. The sequences from the deer BVDV were distinct from those of the bovine type 1a BVDV, but similar to other bovine type 1a BVDVs. Seventy-four (60%) of 124 sera collected from mule deer in this area had serum neutralizing antibody titers to type 1a BVDV of > or = 1:32. The high prevalence of seropositive mule deer and isolation of BVDV suggests that this virus circulates in the mule deer population. The isolate described in this report is the second reported BVDV isolate from free-ranging deer in North America and the first from a mule deer. PMID:11310881

Van Campen, H; Ridpath, J; Williams, E; Cavender, J; Edwards, J; Smith, S; Sawyer, H

2001-04-01

188

Dielectrophoretic isolation and detection of cfc-DNA nanoparticulate biomarkers and virus from blood.  

PubMed

Dielectrophoretic (DEP) microarray devices allow important cellular nanoparticulate biomarkers and virus to be rapidly isolated, concentrated, and detected directly from clinical and biological samples. A variety of submicron nanoparticulate entities including cell free circulating (cfc) DNA, mitochondria, and virus can be isolated into DEP high-field areas on microelectrodes, while blood cells and other micron-size entities become isolated into DEP low-field areas between the microelectrodes. The nanoparticulate entities are held in the DEP high-field areas while cells are washed away along with proteins and other small molecules that are not affected by the DEP electric fields. DEP carried out on 20 ?L of whole blood obtained from chronic lymphocytic leukemia patients showed a considerable amount of SYBR Green stained DNA fluorescent material concentrated in the DEP high-field regions. Whole blood obtained from healthy individuals showed little or no fluorescent DNA materials in the DEP high-field regions. Fluorescent T7 bacteriophage virus could be isolated directly from blood samples, and fluorescently stained mitochondria could be isolated from biological buffer samples. Using newer DEP microarray devices, high-molecular-weight DNA could be isolated from serum and detected at levels as low as 8-16 ng/mL. PMID:23436471

Sonnenberg, Avery; Marciniak, Jennifer Y; McCanna, James; Krishnan, Rajaram; Rassenti, Laura; Kipps, Thomas J; Heller, Michael J

2013-04-01

189

MOLECULAR CHARACTERIZATION OF RABIES VIRUS ISOLATES FROM MEXICO: IMPLICATIONS FOR TRANSMISSION DYNAMICS AND HUMAN RISK  

Microsoft Academic Search

Twenty-eight samples from humans and domestic and wild animals collected in Mexico between 1990 and 1995 were characterized by using anti-nucleoprotein monoclonal antibodies and limited sequence analysis of the nucleoprotein gene. The variants of rabies viruses identified in these samples were compared with other isolates from Mexico and the rest of the Americas to establish epidemiologic links between cases and

CECILIA C. DE MATTOS; CARLOS A. DE MATTOS; ELIZABETH LOZA-RUBIO; ALVARO AGUILAR-SETIEN; LILLIAN A. ORCIARI; JEAN S. SMITH

1999-01-01

190

Complete genome sequence of a bovine viral diarrhea virus strain isolated in southern china.  

PubMed

We report here the full-length RNA genomic sequence of the bovine viral diarrhea virus (BVDV) strain GX4, isolated from a cow in southern China. Studies indicate that BVDV GX4 belongs to the BVDV-1b subtype. This report will help in understanding the epidemiology and molecular characteristics of BVDV in southern China cattle. PMID:24948756

Xie, Zhixun; Fan, Qing; Xie, Zhiqin; Liu, Jiabo; Pang, Yaoshan; Deng, Xianwen; Xie, Liji; Luo, Sisi; Khan, M I

2014-01-01

191

Neuraminidase Sequence Analysis and Susceptibilities of Influenza Virus Clinical Isolates to Zanamivir and Oseltamivir  

Microsoft Academic Search

The influenza virus neuraminidase (NA) inhibitors zanamivir and oseltamivir were introduced into clinical practice in various parts of the world between 1999 and 2002. In order to monitor the potential development of resistance, the Neuraminidase Inhibitor Susceptibility Network was established to coordinate testing of clinical isolates collected through the World Health Organization influenza surveillance network from different regions of the

J. McKimm-Breschkin; T. Trivedi; A. Hampson; A. Hay; A. Klimov; M. Tashiro; F. Hayden; M. Zambon

2003-01-01

192

Complete Nucleotide Sequence of Canine Distemper Virus CDV-PS, Isolated from Dogs in China  

PubMed Central

A new strain of canine distemper virus, CDV-PS, has been isolated from dogs in China, and its complete genome has been sequenced and analyzed. The phylogenetic analysis suggests that CDV-PS belongs to the Asia-1 cluster and has low identity to the vaccine strain. PMID:23682141

Yi, Li; Xu, Hongli; Wang, Jianke; Cheng, Yuening; Zhang, Hailing; Yan, Xijun

2013-01-01

193

Complete Genome Sequences of Two Crimean-Congo Hemorrhagic Fever Viruses Isolated in China  

PubMed Central

Here, we report the complete genome sequences of two Crimean-Congo hemorrhagic fever virus (CCHFV) strains, 79121M18 and YL04057, isolated in Xinjiang, China. Sequence analysis showed that they represent a genotype of CCHFV that has not been reported before. PMID:23908296

Zhou, Zhaorui; Meng, Weiwei; Deng, Fei; Xia, Han; Li, Tianxian; Sun, Surong; Wang, Manli; Wang, Hualin

2013-01-01

194

Avian Influenza Virus with Hemagglutinin-Neuraminidase Combination H8N8, Isolated in Russia  

PubMed Central

We report the genome sequence of an avian influenza virus (AIV) subtype H8N8, isolated in Russia. The genome analysis shows that all genes belong to AIV Eurasian lineages. The PB2 gene was similar to a Mongolian low-pathogenic (LP) AIV H7N1 and a Chinese high-pathogenic (HP) AIV H5N2. PMID:24903874

Sharshov, Kirill A.; Pantin-Jackwood, Mary; Muzyka, Vladimir V.; Shestopalov, Alexander M.

2014-01-01

195

Complete Nucleotide Sequence of Canine Distemper Virus HLJ1-06, Isolated from Foxes in China.  

PubMed

A new strain of canine distemper virus, HLJ1-06, has been isolated from foxes in China, and its complete genome has been sequenced and analyzed. The phylogenetic analysis suggests that HLJ1-06 belongs to the Asia-1 cluster and has low identity to the vaccine strain. PMID:23405331

Jiang, Qian; Hu, Xiaoliang; Ge, Yanhua; Lin, Huan; Jiang, Yong; Liu, Jiasen; Guo, Dongchun; Si, Changde; Qu, Liandong

2013-01-01

196

Molecular characterization of the Great Lakes viral hemorrhagic septicemia virus (VHSV) isolate from USA  

Microsoft Academic Search

BACKGROUND: Viral hemorrhagic septicemia virus (VHSV) is a highly contagious viral disease of fresh and saltwater fish worldwide. VHSV caused several large scale fish kills in the Great Lakes area and has been found in 28 different host species. The emergence of VHS in the Great Lakes began with the isolation of VHSV from a diseased muskellunge (Esox masquinongy) caught

Arun Ammayappan; Vikram N Vakharia

2009-01-01

197

Complete Genome Sequence of Two Middelburg Viruses Isolated from Arthropods in the Central African Republic  

PubMed Central

Arboviral diseases are a major threat to human and animal health today. Analysis of whole-genome sequences of decades-old arboviral strains may bring new insights into the viral evolution that might have facilitated outbreaks. Here, we report the whole-genome sequences of two Middelburg viruses isolated several decades ago in the Central African Republic. PMID:25342688

Berthet, Nicolas; Descorps-Declere, Stephane; Nakoune, Emmanuel; Kazanji, Mirdad

2014-01-01

198

Genetic variability in the coat protein genes of Cymbidium mosaic virus isolates from orchids.  

PubMed

The variability in the nucleotide (nt) and amino acid (aa) sequences of the coat protein (CP) of Cymbidium mosaic virus (CymMV), which naturally infects orchids worldwide, was investigated. The CP genes of 55 CymMV isolates originating from different locations in Korea were amplified using RT-PCR and sequenced. The encoded CP consists of 223 aa. The CP sequences of the Korean isolates were compared with those of previously published CymMV isolates originating from different countries at both nt and aa levels. The Korean isolates shared 74.9-98.3 and 52.7-100% CP homology with CymMV isolates from other countries at the nt and aa levels, respectively. No particular region of variability could be found in either grouping of viruses. In the deduced CymMV CP aa sequence, the C-terminal region was more divergent than the N-terminal. The phylogenetic tree analysis based on nt sequence diversity of CP genes of CymMV isolates supported the hypothesis that CymMV isolates were divided into two subgroups. However, these subgroups were not formed by phylogenetic tree analysis of CP aa sequences. There was no distinct correlation between geographical locations and specific sequence identity, while recombination analysis revealed that there were no intra-specific recombination events among CymMV isolates. PMID:22015427

Yoon, Ju-Yeon; Chung, Bong-Nam; Choi, Gug-Seoun; Choi, Seung-Kook

2012-04-01

199

Complete genomic characterization of a potato mop-top virus isolate from the United States.  

PubMed

Potato mop-top virus (PMTV; family Virgaviridae) was reported recently in the Pacific Northwestern USA. To better understand the genetic diversity of this virus, the complete genome of an isolate from Washington State (WA), USA, was characterized. Sequence comparisons of the WA isolate with other known sequences revealed that the RNA-Rep-encoded RdRp protein and the RNA-CP-encoded coat protein displayed >99 % amino acid sequence identity to those of two Nordic (RdRp) and several European and North American isolates (CP), respectively. The RNA-TGB-encoded TGB 1 and TGB 3 protein sequences had >99 % amino acid sequence identity to the corresponding proteins of Czech and Danish isolates, whereas the TGB 2 protein is identical to those of Colombian isolates. Phylogenetic analysis of the viral genes of the WA isolate reflected the close relationship between WA and European isolates. RFLP analysis of corresponding DNA of RNA TGB and RNA CP revealed that the WA isolate has the RNA TGB-II and RNA CP-B types, which are prevalent in Europe and other parts of world. This is the first report of the complete genome characterization of PMTV from the Americas. PMID:25287129

Ramesh, S V; Raikhy, G; Brown, C R; Whitworth, J L; Pappu, H R

2014-12-01

200

Isolation and Identification of a Novel Rabies Virus Lineage in China with Natural Recombinant Nucleoprotein Gene  

PubMed Central

Rabies virus (RABV) causes severe neurological disease and death. As an important mechanism for generating genetic diversity in viruses, homologous recombination can lead to the emergence of novel virus strains with increased virulence and changed host tropism. However, it is still unclear whether recombination plays a role in the evolution of RABV. In this study, we isolated and sequenced four circulating RABV strains in China. Phylogenetic analyses identified a novel lineage of hybrid origin that comprises two different strains, J and CQ92. Analyses revealed that the virus 3? untranslated region (UTR) and part of the N gene (approximate 500 nt in length) were likely derived from Chinese lineage I while the other part of the genomic sequence was homologous to Chinese lineage II. Our findings reveal that homologous recombination can occur naturally in the field and shape the genetic structure of RABV populations. PMID:23226506

Yan, Hong-Yan; Ding, Nai-Zheng; He, Hong-Bin; Yan, Jia-Xin; Xu, Ge-Lin

2012-01-01

201

Phylogeography of rabies virus isolated from herbivores and bats in the Espírito Santo State, Brazil.  

PubMed

Rabies is enzootic in the State of Espírito Santo, Brazil. Every year, cattle and horses die from rabies that is transmitted by the vampire bat Desmodus rotundus. This paper describes the spread of the rabies virus by the continuous diffusion model using relaxed random walks with BEAST software. Forty-one (41) sequences of gene G from the rabies virus that was isolated from bats and domestic herbivores from several areas of the state between 2006 and 2010 were analyzed. The phylogenetic tree showed three main clusters as well as two sub-clusters under cluster 2. A spatial analysis showed that three strains of the rabies virus spread independently. In general, central Espírito Santo, which is mountainous, was the area where separation of the virus strains occurred. This physical barrier, however, was overcome at some point in time, as samples from different lineages were found in the same microarea. PMID:23264105

Vieira, Luiz Fernando Pereira; Pereira, Sílvia Regina Ferreira Gonçalves; Carnieli, Pedro; Tavares, Luiz Carlos Barbosa; Kotait, Ivanete

2013-04-01

202

Isolation and Characterization of Sylvatic Mosquito-Borne Viruses in Trinidad: Enzootic Transmission and a New Potential Vector of Mucambo Virus  

PubMed Central

Mosquito surveillance was carried out in three forested regions of Trinidad during July 2007–March 2009. A total of 185,397 mosquitoes representing at least 46 species was collected, divided into pools of 1–50 mosquitoes according to species and sex, and screened for arboviruses using cytopathic effect assays on Vero cell monolayers. Eighty-five viruses were isolated, including members of the genera Alphavirus (Mucambo virus; MUCV) and Orthobunyavirus (Caraparu, Oriboca, Bimiti, and Wyeomyia viruses). Species of the Culex subgenus Melanoconion accounted for 56% of the total number of mosquitoes collected and 97% of the viruses isolated; Cx. (Mel.) portesi accounted for 92% of virus isolations. Our results also implicate for the first time Aedes (Ochlerotatus) hortator as a potential vector of MUCV. Phylogenetic analyses of 43 MUCV strains suggest population subdivision within Trinidad, consistent with the hypothesis of enzootic maintenance in localized rodent populations. PMID:21118932

Auguste, Albert J.; Adams, A. Paige; Arrigo, Nicole C.; Martinez, Raymond; Travassos da Rosa, Amelia P. A.; Adesiyun, Abiodun A.; Chadee, Dave D.; Tesh, Robert B.; Carrington, Christine V. F.; Weaver, Scott C.

2010-01-01

203

Isolation and Characterization of a Single-Stranded DNA Virus Infecting Chaetoceros lorenzianus Grunow?  

PubMed Central

Diatoms are one of the most significant primary producers in the ocean, and the importance of viruses as a potential source of mortality for diatoms has recently been recognized. Thus far, eight different diatom viruses infecting the genera Rhizosolenia and Chaetoceros have been isolated and characterized to different extents. We report the isolation of a novel diatom virus (ClorDNAV), which causes the lysis of the bloom-forming species Chaetoceros lorenzianus, and show its physiological, morphological, and genomic characteristics. The free virion was estimated to be ?34 nm in diameter. The arrangement of virus particles appearing in cross-section was basically a random aggregation in the nucleus. Occasionally, distinctive formations such as a ring-like array composed of 9 or 10 spherical virions or a centipede-like array composed of rod-shaped particles were also observed. The latent period and the burst size were estimated to be <48 h and 2.2 × 104 infectious units per host cell, respectively. ClorDNAV harbors a covalently closed circular single-stranded DNA (ssDNA) genome (5,813 nucleotides [nt]) that includes a partially double-stranded DNA region (979 nt). At least three major open reading frames were identified; one showed a high similarity to putative replicase-related proteins of the other ssDNA diatom viruses, Chaetoceros salsugineum DNA virus (previously reported as CsNIV) and Chaetoceros tenuissimus DNA virus. ClorDNAV is the third member of the closed circular ssDNA diatom virus group, the genus Bacilladnavirus. PMID:21666026

Tomaru, Yuji; Takao, Yoshitake; Suzuki, Hidekazu; Nagumo, Tamotsu; Koike, Kanae; Nagasaki, Keizo

2011-01-01

204

Antigenic typing of Brazilian rabies virus samples isolated from animals and humans, 1989-2000.  

PubMed

Animal and human rabies samples isolated between 1989 and 2000 were typified by means of a monoclonal antibody panel against the viral nucleoprotein. The panel had been previously established to study the molecular epidemiology of rabies virus in the Americas. Samples were isolated in the Diagnostic Laboratory of the Pasteur Institute and in other rabies diagnostic centers in Brazil. In addition to the fixed virus samples CVS-31/96-IP, preserved in mouse brain, and PV-BHK/97, preserved in cell culture, a total of 330 rabies virus samples were isolated from dogs, cats, cattle, horses, bats, sheep, goat, swine, foxes, marmosets, coati and humans. Six antigenic variants that were compatible with the pre-established monoclonal antibodies panel were defined: numbers 2 (dog), 3 (Desmodus rotundus), 4 (Tadarida brasiliensis), 5 (vampire bat from Venezuela), 6 (Lasiurus cinereus) and Lab (reacted to all used antibodies). Six unknown profiles, not compatible with the panel, were also found. Samples isolated from insectivore bats showed the greatest variability and the most commonly isolated variant was variant-3 (Desmodus rotundus). These findings may be related to the existence of multiple independent transmission cycles, involving different bat species. PMID:12048546

Favoretto, Silvana Regina; Carrieri, Maria Luiza; Cunha, Elenice Maria S; Aguiar, Elizabeth A C; Silva, Luzia Helena Q; Sodre, Miriam M; Souza, Maria Conceiçăo A M; Kotait, Ivanete

2002-01-01

205

First Report of Cucumber mosaic virus Isolated from Wild Vigna angularis var. nipponensis in Korea  

PubMed Central

A viral disease causing severe mosaic, necrotic, and yellow symptoms on Vigna angularis var. nipponensis was prevalent around Suwon area in Korea. The causal virus was characterized as Cucumber mosaic virus (CMV) on the basis of biological and nucleotide sequence properties of RNAs 1, 2 and 3 and named as CMV-wVa. CMV-wVa isolate caused mosaic symptoms on indicator plants, Nicotiana tabacum cv. Xanthi-nc, Petunia hybrida, and Cucumis sativus. Strikingly, CMV-wVa induced severe mosaic and malformation on Cucurbita pepo, and Solanum lycopersicum. Moreover, it caused necrotic or mosaic symptoms on V. angularis and V. radiate of Fabaceae. Symptoms of necrotic local or pin point were observed on inoculated leaves of V. unguiculata, Vicia fava, Pisum sativum and Phaseolus vulgaris. However, CMV-wVa isolate failed to infect in Glycine max cvs. ‘Sorok’, ‘Sodam’ and ‘Somyeong’. To assess genetic variation between CMV-wVa and the other known CMV isolates, phylogenetic analysis using 16 complete nucleotide sequences of CMV RNA1, RNA2, and RNA3 including CMV-wVa was performed. CMV-wVa was more closely related to CMV isolates belonging to CMV subgroup I showing about 85.1–100% nucleotide sequences identity to those of subgroup I isolates. This is the first report of CMV as the causal virus infecting wild Vigna angularis var. nipponensis in Korea. PMID:25289004

Kim, Mi-Kyeong; Jeong, Rae-Dong; Kwak, Hae-Ryun; Lee, Su-Heon; Kim, Jeong-Soo; Kim, Kook-Hyung; Cha, Byeongjin; Choi, Hong-Soo

2014-01-01

206

Genetic variation in potato virus M isolates infecting pepino (Solanum muricatum) in China.  

PubMed

Potato virus M (PVM, genus Carlavirus, family Betaflexviridae) is considered to be one of most economically important pathogens of pepino in China. However, the details and the mechanisms underlying PVM evolution are unknown. In this study, we determined and analyzed 40 TGB 1 gene sequences, 67 TGB 2 and TGB 3 gene sequences, and 88 CP and NABP gene sequences from viruses isolated from 19 samples of pepino (Solanum muricatum) and one sample of tomato (S. lycopersicum) collected from different areas of China. Recombination analysis identified only one clear recombinant in the TGB2-TGB3-CP region, but no recombinants were detected for each of the five individual genes. Phylogenetic analysis showed that all PVM isolates could be divided into at least two lineages in trees derived from the TGB 2, CP, and NABP gene sequences, and the lineages seemed to reflect geographical origin. The five PVM genes in this study were found to be under strong negative selection pressure. The PVM isolates examined showed frequent gene flow between the Chinese and European populations, and also within the Chinese population. Clear star phylogenies and the neutral equilibrium model test showed that pepino isolates of PVM appear to be experiencing a new expansion after a recent introduction into China, and these isolates display low levels of genetic diversity. To our knowledge, this study is the first report describing genetic structure, recombination, and gene flow in PVM populations, and it provides strong evolutionary evidence for the virus populations from different geographic regions of China. PMID:25233939

Ge, Beibei; He, Zhen; Zhang, Zhixiang; Wang, Hongqing; Li, Shifang

2014-12-01

207

Isolation of an Hswl Nav4 influenza virus from a tufted duck (Aythya fuligula) in Japan.  

PubMed

A hemagglutinating agent was isolated from a tufted duck captured in Lake Shinji, Shimane Prefecture, Japan, and identified as influenza A by double immunodiffusion tests with the antiserum to influenza A virus ribonucleoprotein. The hemagglutinin in the isolate was antigenically related to that of A/New Jersey/8/76 but was not identical with it. The neuraminidase antigen of this Hswl subtype was closely related to that of A/turkey/Ontario/6118/68 and was shown to be Nav4 subtype. After experimental infection of 5-week-old SPF-chickens with the isolate, virus was recovered from various organs including the brain, despite the absence of signs of disease. PMID:6270508

Tsubokura, M; Otsuki, K; Yamamoto, H; Kawaoka, Y; Nerome, K

1981-01-01

208

Isolation and characterization of caprine arthritis encephalitis virus in goats from Poland.  

PubMed

The caprine arthritis-encephalitis virus (CAEV) was isolated from monocyte-derived macrophages (M/M), but not from PBMC of seropositive goats by co-cultivation with goat synovial membrane cells. Out of eight M/M co-cultures, CAEV was evidenced by the syncytia formation and presence of proviral DNA in two and four cultures, respectively. Two virus isolates from co-cultures showing cytopathic effects were further confirmed as CAEV by western blotting, PCR, and sequence analysis. The nucleotide sequence of gag gene showed 92.0% and 90.3% homology to the prototype CAEV-Co strain. Supernatants harvested from these cultures induced syncytia when cultured with uninfected cells and the resultant titer was 10(3.5) and 10(2.5) TCID50 per ml. New CAEV isolates are suitable candidates for further analysis of their genetic and biological properties. PMID:19645347

Kaba, J; Rola, M; Materniak, M; Ku?mak, J; Nowicki, M

2009-01-01

209

Studies on Bunyaviridae including Zaliv Terpeniya virus isolated from Ixodes uriae ticks (Acarina: Ixodidae) in Brittany, France.  

PubMed

Three viruses were isolated from ticks parasitizing nesting seabirds on Cape Sizun, Brittany, France, during the spring of 1979: Soldado like virus (Bunyaviridae, Nairovirus genus) from Ornithodoros (Alectorobius) maritimus (Argasidae), Zaliv Terpeniya (ZT) virus (Bunyaviridae, Uukuvirus genus) from Ixodes (Ceratixodes) uriae (Ixodidae) parasitizing chicks of the kittiwake (Rissa tridactyla), and a virus of the Sakhalin group from I. (C.) uriae. Certain virological and ultrastructural properties of ZT virus were studied and the antigenic relationships between th French isolate and other UUK serogroup viruses from I. (C.) uriae were analyzed in the complement fixation and immune diffusion tests. Cape Sizun is notable for the unusual presence of both argasied (Ornithodoros) and ixodid (Ixodes) ticks infesting seabirds in adjacent niches in a single locality, where they are infected by different viruses. PMID:6173027

Chastel, C; Monnat, J Y; Le Lay, G; Guiguen, C; Quillien, M C; Beaucournu, J C

1981-01-01

210

Complete genome analysis of three isolates of narcissus late season yellows virus and two of narcissus yellow stripe virus: three species or one?  

PubMed

Complete genome sequences of two new isolates of narcissus late season yellows virus (NLSYV) from Australia were compared with the other NLSYV genome from China and with two complete genomes of isolates designated narcissus yellow stripe virus (NYSV), one from Australia and the other from China. On the basis of symptoms on natural and experimental host species, and genome sequence identity, the isolates could either be classified as closely related members of three different species or placed together in one taxon. Options for classification of these potyvirus isolates are discussed. PMID:24385160

Wylie, Stephen J; Li, Hua; Sivasithamparam, Krishnapillai; Jones, Michael G K

2014-06-01

211

A comparative study of rabies virus isolates from hematophagous bats in Brazil.  

PubMed

The Brazilian chiropteran fauna consists of 167 species; of which, three are hematophagous: the common vampire bat (Desmodus rotundus), the white-winged vampire bat (Diaemus youngi), and the hairy-legged vampire bat (Diphylla ecaudata). The aim of this study was to describe the isolation of Rabies virus from common and hairy-legged vampire bats and to report the first comparative antigenic and genetic studies of isolates from these bats. Antigenic and genetic typing of both isolates identified them as antigenic variant 3 (AgV3), the variant frequently isolated from common vampire bats. Phylogenetic analysis showed 99.3% identity between the isolates. This is the first time since 1934 that Rabies virus has been isolated from hairy-legged vampire bats in Brazil. Our analysis provides evidence that the existence of rabies-positive isolates from hairy-legged vampire bats may be the result of an interspecific rabies transmission event from common vampire bats and suggests that roost cohabitation may occur. PMID:20966291

Castilho, Juliana G; Carnieli, Pedro; Oliveira, Rafael N; Fahl, Willian O; Cavalcante, Rosangela; Santana, Antonio A; Rosa, Wellington L G A; Carrieri, Maria L; Kotait, Ivanete

2010-10-01

212

Development of FPV140 antigen-specific ELISA differentiating fowlpox virus isolates from all other viral pathogens of avian origin.  

PubMed

The FPV140 gene encodes an envelope protein of fowlpox virus (FPV). In this study, the FPV140 gene of FPV Chinese isolate HH2008 was cloned and the comparison of its sequence with other FPV isolates showed it to be highly conserved across all FPV isolates. A recombinant plasmid pET-FPV140 carrying FPV140 gene was constructed and transformed into Escherichia coli. The optimal expression condition for the FPV140 gene was developed and purified FPV140 recombinant protein was used to produce rabbit polyclonal antibody. An indirect ELISA using this anti-FPV140 polyclonal antibody was capable of distinguishing avian FPV isolates from other common avian pathogens such as mycoplasma gallisepticum, infectious laryngotracheitis virus, avian influenza virus, infectious bursal disease virus, and avian infectious bronchitis virus. This ELISA will serve as a useful diagnostic tool for the detection of FPV in clinical samples. PMID:22991535

Li, G; Hong, Q; Ren, Y; Lillehoj, H S; He, C; Ren, X

2012-10-01

213

Molecular characterization of the VP2 gene of infectious pancreatic necrosis virus (IPNV) isolates from Mexico.  

PubMed

Infectious pancreatic necrosis virus (IPNV) is one of the most important viruses in the Pacific salmon Oncorhynchus spp., Atlantic Salmon Salmo salar, and Rainbow Trout O. mykiss industry. This virus has been shown to produce high mortality among salmonid fry and juveniles, and survivors might become carriers. Since 2000, IPNV has affected Mexican Rainbow Trout culture, resulting in considerable economic losses. In the current study, molecular characterization of the VP2 gene of a number of Mexican IPNV isolates was done and the virus's phylogenetic relationships to IPNV reference strains were investigated. The phylogenetic analysis indicated that Mexican IPNV isolates are closely related to strains from the United States and Canada and that all Mexican IPNV isolates belong to genogroup 1. Furthermore, low genetic diversity was found between the Mexican isolates (identity, 95.8-99.8% nucleotides and 95.8-99.6% amino acids). The result of the analysis of the amino acid residues found at positions 217, 221, and 247 (alanine, threonine, and glutamic acid, respectively) could be associated with virulence, although the expression of virulence factors is more complex and may be influenced by the agent and host factors. The high percentage of identity among the VP2 genes from geographically distant IPNV isolates and the evidence of wide distribution in the country might have been facilitated by carrier trout. This hypothesis is supported by the identification of the amino acid threonine at position 221 in all Mexican isolates, a factor related to the carrier state for IPNV, as reported by other studies. PMID:24689957

Salgado-Miranda, Celene; Rojas-Anaya, Edith; García-Espinosa, Gary; Loza-Rubio, Elizabeth

2014-03-01

214

Properties of a virus isolated from Vernonia amygdalina Del. in Lagos, Nigeria.  

PubMed

A previously uncharacterized virus tentatively named Vernonia green vein-banding virus (VGVBV) was isolated from Vernonia amygdalina Del. ("bitterleaf") from Lagos, Nigeria. The virus was mechanically transmissible but had a narrow host range restricted to Nicotiana benthamiana, Chenopodium quinoa and C. amaranticolor. It was also transmissible in a non-persistent manner by Myzus persicae. The virus was purified from N. benthamiana and about 750 nm long flexuous rod-shaped particles were observed in purified preparations as well as in leaf-dips of Vernonia sp. Inclusion bodies in the form of pinwheels and scrolls were observed in ultrathin sections of Vernonia leaves by electron microscopy. M(r), of the viral coat protein was estimated to be about 34 K. In indirect ELISA, all 20 samples from naturally infected Vernonia sp. reacted positively with a potyvirus-specific monoclonal antibody (MAb) as well as with an antiserum raised against VGVBV. Apart from the homologous antigen, the VGVBV antiserum reacted only with Plum poxvirus (PPV). The VGVBV reacted strongly with the antisera to Bean yellow mosaic virus (BYMV), Bean common mosaic virus (BCMV) and Amaranthus leaf mottle virus (AmLMV) but weakly with antisera to PPV and Cowpea aphid-borne mosaic virus (CABMV) (all members of the family Potyviridae, the genus Potyvirus) in at least one of the assays used (indirect ELISA, dot-blot immunoassay and Western blot analysis). The results of our host range, cytopathological and serological studies and the available literature indicate that a hitherto difficult to transmit VGVBV has only been reported from Nigeria. We consider VGVBV a candidate for a new potyvirus. This virus should be further investigated to collect sufficient data for a qualified proposal of VGVBV as a new potyvirus. PMID:15745049

Taiwo, M A; Dijkstra, J

2004-01-01

215

Viral Replication, Persistence in Water and Genetic Characterization of Two Influenza A Viruses Isolated from Surface Lake Water  

PubMed Central

Water-borne transmission has been suggested as an important transmission mechanism for Influenza A (IA) viruses in wild duck populations; however, relatively few studies have attempted to detect IA viruses from aquatic habitats. Water-isolated viruses have rarely been genetically characterized and evaluation for persistence in water and infectivity in natural hosts has never been documented. In this study, we focused on two IA viruses (H3N8 and H4N6 subtypes) isolated from surface lake water in Minnesota, USA. We investigated the relative prevalence of the two virus subtypes in wild duck populations at the sampling site and their genetic relatedness to IA viruses isolated in wild waterbirds in North America. Viral persistence under different laboratory conditions (temperature and pH) and replication in experimentally infected Mallards (Anas platyrhynchos) were also characterized. Both viruses were the most prevalent subtype one year following their isolation in lake water. The viruses persisted in water for an extended time period at constant temperature (several weeks) but infectivity rapidly reduced under multiple freeze-thaw cycles. Furthermore, the two isolates efficiently replicated in Mallards. The complete genome characterization supported that these isolates originated from genetic reassortments with other IA viruses circulating in wild duck populations during the year of sampling. Based on phylogenetic analyses, we couldn't identify genetically similar viruses in duck populations in the years following their isolation from lake water. Our study supports the role for water-borne transmission for IA viruses but also highlights that additional field and experimental studies are required to support inter-annual persistence in aquatic habitats. PMID:22028909

Lebarbenchon, Camille; Yang, My; Keeler, Shamus P.; Ramakrishnan, Muthannan A.; Brown, Justin D.; Stallknecht, David E.; Sreevatsan, Srinand

2011-01-01

216

A bovine viral diarrhea virus type 1a strain in China: isolation, identification, and experimental infection in calves  

PubMed Central

Background Bovine viral diarrhea virus (BVDV) is one of the most important pathogens in cattle. Previously, BVDV sub-genotypes of 1b, 1c, 1d, and 1 m were detected in China. However, isolation of BVDV type 1a from cattle has not been reported in China. In 2010, twenty nasal swabs and blood samples were collected from the cattle suspected BVDV infection in Henan province, China. A BVDV isolate was isolated using cell culture, and the pathogenesis of the virus isolate was studied. Methods Virus isolation was performed on MDBK cells. The virus identification was conducted by RT-PCR, neutralization test and immunofluorescence assay. In order to determine the genotype of the newly isolated virus, the 5? un-translated region (5?UTR) of the virus isolate was cloned, sequenced and phylogenetically analyzed. To evaluate the virulence of the virus isolate, four BVDV sero-negative calves were intranasally inoculated with the virus suspension. Rectal temperatures and clinical signs were recorded daily. Blood samples were analyzed for changes in white blood cell counts, and tissue samples were taken for histopathology analysis. Results A new isolate of bovine viral diarrhea virus (BVDV), named HN01, was isolated from the nasal swabs using MDBK cell culture. The HN01 strain caused cytopathic effect (CPE) in MDBK cell cultures after two passages. The virus specifically reacted to BVDV1-specific monoclonal antibody in an immunofluorescence assay. A fragment of 288 bp of genome from this isolate was amplified by the RT-PCR. Phylogenetic analysis of 5?UTR indicated that the virus was BVDV 1a. In the pathogenesis study, four calves experimentally infected with the BVDV strain developed depression, cough and other clinical signs. Calves showed high temperature over 40°C, and white blood cell counts dropped more than 40%. Conclusions A new subgenotype 1a strain of BVDV was firstly isolated from dairy cattle in China. The experimental infection showed that the virus was moderate pathogenic to cattle and can be used as a BVDV challenge virus to evaluate the efficacy of BVDV vaccines in the target animals. PMID:24444389

2014-01-01

217

Characterization of herpes simplex virus clinical isolate Y3369 as a glycoprotein G variant and its bearing on virus typing  

PubMed Central

Background Herpes simplex viruses exist as two major serotypes, type 1 (HSV-1) and type 2 (HSV-2). Determination of type, either HSV-1 or HSV-2, is important in accurate diagnosis and clinical control of transmission. Several tests are available for typing HSV, including a monoclonal antibody specific for glycoprotein G and several PCR assays. Findings A clinical isolate was identified as herpes simplex virus, but tested negative for both HSV-1 and HSV-2 antigens using type-specific monoclonal antibody assays. The isolate was determined to be HSV-1 by PCR analysis. A mutation which likely caused the monoclonal antibody non-reactivity was found in glycoprotein G. Phylogenetic analysis revealed two groups of HSV, one with the mutation and one without. Three population studies examining mutations in HSV-1 glycoprotein G were analyzed by chi-squared test. To this point, the epitope which the monoclonal antibody recognizes was only found in HSV-1 isolates from human European populations (p < 0.0001). Conclusions These findings suggest that the PCR-based methods for HSV typing may be more useful than the standard monoclonal antibody test in areas of the world where the variant in glycoprotein G is more prevalent. PMID:21658271

2011-01-01

218

Phospholipase A2 Isolated from the Venom of Crotalus durissus terrificus Inactivates Dengue virus and Other Enveloped Viruses by Disrupting the Viral Envelope  

PubMed Central

The Flaviviridae family includes several virus pathogens associated with human diseases worldwide. Within this family, Dengue virus is the most serious threat to public health, especially in tropical and sub-tropical regions of the world. Currently, there are no vaccines or specific antiviral drugs against Dengue virus or against most of the viruses of this family. Therefore, the development of vaccines and the discovery of therapeutic compounds against the medically most important flaviviruses remain a global public health priority. We previously showed that phospholipase A2 isolated from the venom of Crotalus durissus terrificus was able to inhibit Dengue virus and Yellow fever virus infection in Vero cells. Here, we present evidence that phospholipase A2 has a direct effect on Dengue virus particles, inducing a partial exposure of genomic RNA, which strongly suggests inhibition via the cleavage of glycerophospholipids at the virus lipid bilayer envelope. This cleavage might induce a disruption of the lipid bilayer that causes a destabilization of the E proteins on the virus surface, resulting in inactivation. We show by computational analysis that phospholipase A2 might gain access to the Dengue virus lipid bilayer through the pores found on each of the twenty 3-fold vertices of the E protein shell on the virus surface. In addition, phospholipase A2 is able to inactivate other enveloped viruses, highlighting its potential as a natural product lead for developing broad-spectrum antiviral drugs. PMID:25383618

Muller, Vanessa Danielle; Soares, Ricardo Oliveira; dos Santos-Junior, Nilton Nascimento; Trabuco, Amanda Cristina; Cintra, Adelia Cristina; Figueiredo, Luiz Tadeu; Caliri, Antonio; Sampaio, Suely Vilela; Aquino, Victor Hugo

2014-01-01

219

First isolation of dengue virus from the 2010 epidemic in Nepal.  

PubMed

Dengue is an emerging disease in Nepal and was first observed as an outbreak in nine lowland districts in 2006. In 2010, however, a large epidemic of dengue occurred with 4,529 suspected and 917 serologically-confirmed cases and five deaths reported in government hospitals in Nepal. The collection of demographic information was performed along with an entomological survey and clinical evaluation of the patients. A total of 280 serum samples were collected from suspected dengue patients. These samples were subjected to routine laboratory investigations and IgM-capture ELISA for dengue serological identification, and 160 acute serum samples were used for virus isolation, RT-PCR, sequencing and phylogenetic analysis. The results showed that affected patients were predominately adults, and that 10% of the cases were classified as dengue haemorrhagic fever/ dengue shock syndrome. The genetic characterization of dengue viruses isolated from patients in four major outbreak areas of Nepal suggests that the DENV-1 strain was responsible for the 2010 epidemic. Entomological studies identified Aedes aegypti in all epidemic areas. All viruses belonged to a monophyletic single clade which is phylogenetically close to Indian viruses. The dengue epidemic started in the lowlands and expanded to the highland areas. To our knowledge, this is the first dengue isolation and genetic characterization reported from Nepal. PMID:24155651

Pandey, Basu D; Nabeshima, Takeshi; Pandey, Kishor; Rajendra, Saroj P; Shah, Yogendra; Adhikari, Bal R; Gupta, Govinda; Gautam, Ishan; Tun, Mya M N; Uchida, Reo; Shrestha, Mahendra; Kurane, Ichiro; Morita, Kouichi

2013-09-01

220

First Isolation of Dengue Virus from the 2010 Epidemic in Nepal  

PubMed Central

Dengue is an emerging disease in Nepal and was first observed as an outbreak in nine lowland districts in 2006. In 2010, however, a large epidemic of dengue occurred with 4,529 suspected and 917 serologically-confirmed cases and five deaths reported in government hospitals in Nepal. The collection of demographic information was performed along with an entomological survey and clinical evaluation of the patients. A total of 280 serum samples were collected from suspected dengue patients. These samples were subjected to routine laboratory investigations and IgM-capture ELISA for dengue serological identification, and 160 acute serum samples were used for virus isolation, RT-PCR, sequencing and phylogenetic analysis. The results showed that affected patients were predominately adults, and that 10% of the cases were classified as dengue haemorrhagic fever/ dengue shock syndrome. The genetic characterization of dengue viruses isolated from patients in four major outbreak areas of Nepal suggests that the DENV-1 strain was responsible for the 2010 epidemic. Entomological studies identified Aedes aegypti in all epidemic areas. All viruses belonged to a monophyletic single clade which is phylogenetically close to Indian viruses. The dengue epidemic started in the lowlands and expanded to the highland areas. To our knowledge, this is the first dengue isolation and genetic characterization reported from Nepal. PMID:24155651

Pandey, Basu D.; Nabeshima, Takeshi; Pandey, Kishor; Rajendra, Saroj P.; Shah, Yogendra; Adhikari, Bal R.; Gupta, Govinda; Gautam, Ishan; Tun, Mya M. N.; Uchida, Reo; Shrestha, Mahendra; Kurane, Ichiro; Morita, Kouichi

2013-01-01

221

Isolation of avian influenza H5N1 virus from vaccinated commercial layer flock in Egypt  

PubMed Central

Background Uninterrupted transmission of highly pathogenic avian influenza virus (HPAIV) H5N1 of clade 2.2.1 in Egypt since 2006 resulted in establishment of two main genetic clusters. The 2.2.1/C group where all recent human and majority of backyard origin viruses clustered together, meanwhile the majority of viruses derived from vaccinated poultry in commercial farms grouped in 2.2.1.1 clade. Findings In the present investigation, an HPAIV H5N1 was isolated from twenty weeks old layers chickens that were vaccinated with a homologous H5N1 vaccine at 1, 7 and 16 weeks old. At twenty weeks of age, birds showed cyanosis of comb and wattle, decrease in egg production and up to 27% mortality. Examined serum samples showed low antibody titer in HI test (Log2 3.2± 4.2). The hemagglutinin (HA) and neuraminidase (NA) genes of the isolated virus were closely related to viruses in 2.2.1/C group isolated from poultry in live bird market (LBM) and backyards or from infected people. Conspicuous mutations in the HA and NA genes including a deletion within the receptor binding domain in the HA globular head region were observed. Conclusions Despite repeated vaccination of layer chickens using a homologous H5N1 vaccine, infection with HPAIV H5N1 resulted in significant morbidity and mortality. In endemic countries like Egypt, rigorous control measures including enforcement of biosecurity, culling of infected birds and constant update of vaccine virus strains are highly required to prevent circulation of HPAIV H5N1 between backyard birds, commercial poultry, LBM and humans. PMID:23185975

2012-01-01

222

Molecular characterization of two prunus necrotic ringspot virus isolates from Canada.  

PubMed

We determined the entire RNA1, 2 and 3 sequences of two prunus necrotic ringspot virus (PNRSV) isolates, Chr3 from cherry and Pch12 from peach, obtained from an orchard in the Niagara Fruit Belt, Canada. The RNA1, 2 and 3 of the two isolates share nucleotide sequence identities of 98.6%, 98.4% and 94.5%, respectively. Their RNA1- and 2-encoded amino acid sequences are about 98% identical to the corresponding sequences of a cherry isolate, CH57, the only other PNRSV isolate with complete RNA1 and 2 sequences available. Phylogenetic analysis of the coat protein and movement protein encoded by RNA3 of Pch12 and Chr3 and published PNRSV isolates indicated that Chr3 belongs to the PV96 group and Pch12 belongs to the PV32 group. PMID:22327390

Cui, Hongguang; Hong, Ni; Wang, Guoping; Wang, Aiming

2012-05-01

223

A molecular taxonomy for cricket paralysis virus including two new isolates from Australian populations of Drosophila (Diptera: Drosophilidae)  

Microsoft Academic Search

Summary Two new isolates of cricket paralysis virus, TAR and SIM, are described that were originally isolated from laboratory colonies ofDrosophila melanogaster andDrosophila simulans respectively. Using a combination of biological, serological and molecular characters it was possible to distinguish the SIM isolate from all other isolates and it is thus described as a new strain; CrPVSIM. The TAR isolate however,

K. N. Johnson; P. D. Christian

1996-01-01

224

Complete genomic sequence of a border disease virus isolated from Pyrenean chamois.  

PubMed

This report describes the full-length genome sequence of the pestivirus strain H2121 which was recently isolated from Pyrenean chamois and typed as Border disease virus (BDV) genotype 4. Comparison with full-length genomic sequences of the approved pestivirus species Bovine viral diarrhea virus-1 (BVDV-1), BVDV-2, BDV, and Classical swine fever virus, the tentative species represented by strain Giraffe-1, as well as the atypical pestivirus strain Th/04_KhonKaen confirmed that the chamois pestivirus strain is most similar to BDV. The viral genome of H2121 is 12,305 nucleotides long and contains one large open reading frame. The latter encodes a polyprotein consisting of 3899 amino acids and is flanked with 376 nucleotides long 5' untranslated region (UTR) and 229 nt long 3' UTR. The genome organization of the chamois virus is reminiscent to that of other pestiviruses. Compared to other BDV strains including BDV-1 strain X818 and BDV-2 strain Reindeer-1, the 5' UTR and ORF of the chamois virus are very similar in length, while the 3' UTR of H2121 is 31-44 nucleotides shorter. In contrast to other BDV strains, the genome of the chamois virus contains a unique four amino acid insertion at the N-terminus of NS2. PMID:20638945

Vilcek, Stefan; Willoughby, Kim; Nettleton, Peter; Becher, Paul

2010-09-01

225

Endogenous New World primate type C viruses isolated from owl monkey (Aotus trivirgatus) kidney cell line.  

PubMed Central

A type C virus (OMC-1) detected in a culture of owl monkey kidney cells resembled typical type C viruses morphologically, but was slightly larger than previously characterized mammalian type C viruses. OMC-1 can be transmitted to bat lung cells and cat embryo fibroblasts. The virions band at a density of 1.16 g/ml in isopycnic sucrose density gradients and contain reverse transcriptase and a 60-65S RNA genome composed of approximately 32S subunits. The reverse transcriptase is immunologically and biochemically distinct from the polymerases of othe retroviruses. Radioimmunoassays directed to the interspecies antigenic determinants of the major structure proteins of other type C viruses do not detect a related antigen in OMC-1. Nucleic acid hybridization experiments using labeled viral genomic RNA or proviral cDNA transcripts to normal cellular DNA of different species show that OMC-1 is an endogenous virus with multiple virogene copies (20-50 per haploid genome) present in normal owl monkey cells and is distinct from previously isolated type C and D viruses. Sequences related to the OMC-1 genome can be detected in other New World monkeys. Thus, similar to the Old World primates (e.g., baboons as a prototype), the New World monkeys contain endogenous type C viral genes that appear to have been transmitted in the primate germ line. Images PMID:76312

Todaro, G J; Sherr, C J; Sen, A; King, N; Daniel, M D; Fleckenstein, B

1978-01-01

226

Structure of a Venezuelan equine encephalitis virus assembly intermediate isolated from infected cells  

SciTech Connect

Venezuelan equine encephalitis virus (VEEV) is a prototypical enveloped ssRNA virus of the family Togaviridae. To better understand alphavirus assembly, we analyzed newly formed nucleocapsid particles (termed pre-viral nucleocapsids) isolated from infected cells. These particles were intermediates along the virus assembly pathway, and ultimately bind membrane-associated viral glycoproteins to bud as mature infectious virus. Purified pre-viral nucleocapsids were spherical with a unimodal diameter distribution. The structure of one class of pre-viral nucleocapsids was determined with single particle reconstruction of cryo-electron microscopy images. These studies showed that pre-viral nucleocapsids assembled into an icosahedral structure with a capsid stoichiometry similar to the mature nucleocapsid. However, the individual capsomers were organized significantly differently within the pre-viral and mature nucleocapsids. The pre-viral nucleocapsid structure implies that nucleocapsids are highly plastic and undergo glycoprotein and/or lipid-driven rearrangements during virus self-assembly. This mechanism of self-assembly may be general for other enveloped viruses.

Lamb, Kristen; Lokesh, G.L. [Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555-0647 (United States); Sherman, Michael [Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555-0647 (United States); W.M. Keck Center for Virus Imaging, University of Texas Medical Branch, Galveston, TX 77555-0647 (United States); Watowich, Stanley, E-mail: watowich@xray.utmb.ed [Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555-0647 (United States); W.M. Keck Center for Virus Imaging, University of Texas Medical Branch, Galveston, TX 77555-0647 (United States)

2010-10-25

227

Receptor Specificity of Influenza A H3N2 Viruses Isolated in Mammalian Cells and Embryonated Chicken Eggs ? †  

PubMed Central

Isolation of human subtype H3N2 influenza viruses in embryonated chicken eggs yields viruses with amino acid substitutions in the hemagglutinin (HA) that often affect binding to sialic acid receptors. We used a glycan array approach to analyze the repertoire of sialylated glycans recognized by viruses from the same clinical specimen isolated in eggs or cell cultures. The binding profiles of whole virions to 85 sialoglycans on the microarray allowed the categorization of cell isolates into two groups. Group 1 cell isolates displayed binding to a restricted set of ?2-6 and ?2-3 sialoglycans, whereas group 2 cell isolates revealed receptor specificity broader than that of their egg counterparts. Egg isolates from group 1 showed binding specificities similar to those of cell isolates, whereas group 2 egg isolates showed a significantly reduced binding to ?2-6- and ?2-3-type receptors but retained substantial binding to specific O- and N-linked ?2-3 glycans, including ?2-3GalNAc and fucosylated ?2-3 glycans (including sialyl Lewis x), both of which may be important receptors for H3N2 virus replication in eggs. These results revealed an unexpected diversity in receptor binding specificities among recent H3N2 viruses, with distinct patterns of amino acid substitution in the HA occurring upon isolation and/or propagation in eggs. These findings also suggest that clinical specimens containing viruses with group 1-like receptor binding profiles would be less prone to undergoing receptor binding or antigenic changes upon isolation in eggs. Screening cell isolates for appropriate receptor binding properties might help focus efforts to isolate the most suitable viruses in eggs for production of antigenically well-matched influenza vaccines. PMID:20519409

Stevens, James; Chen, Li-Mei; Carney, Paul J.; Garten, Rebecca; Foust, Angie; Le, Jianhua; Pokorny, Barbara A.; Manojkumar, Ramanunninair; Silverman, Jeanmarie; Devis, Rene; Rhea, Karen; Xu, Xiyan; Bucher, Doris J.; Paulson, James; Cox, Nancy J.; Klimov, Alexander; Donis, Ruben O.

2010-01-01

228

Genetic and phylogenetic analysis of South Korean sacbrood virus isolates from infected honey bees (Apis cerana).  

PubMed

Sacbrood virus (SBV) is one of the most destructive honey bee viruses. The virus causes failure to pupate and death in both larvae and adult bees. Genetic analysis of SBV infected honey bees (Apis cerana) from five different provinces was carried out based on three nucleotide sequences; one partial structural protein coding sequence and two non-structural protein coding sequences. Sequences amplified by three specific primer pairs were aligned and compared with reference sequences deposited in the GenBank database. Sequence alignments revealed a low level of sequence variation among Korean isolates (? 98.6% nucleotide identity), regardless of the genome regions studied or the geographic origins of the strains. Multiple sequence comparisons indicated that Korean SBV isolates are genetically closely related to Chinese and other Asian strains. Interestingly, the Korean SBV isolates showed a number of unique nucleotides and amino acids that had not been observed in other published strains. Korean and other Asian isolates from the host A. cerana and the UK, European and Japanese strains from the host Apis mellifera showed differences in nucleotide and deduced amino acid identities. This suggests that host-specificity exists among SBV strains isolated from different species. Phylogenetic relatedness between compared sequences was analyzed by MEGA 4.1 software using the neighbor-joining (NJ) method with a boot-strap value of 1000 replicates. Obtained topologies were in agreement with previous studies, in which a distinct group of SBV was formed by UK and European genotypes and another group was comprised of Asian genotypes including strains that originated from China, Japan (japonica), India and Nepal. However, phylogeny based on a partial protein structural coding sequence grouped all Korean SBV isolates identified in A. cerana as a separate cluster. Our findings suggest that further study, including Korean SBV isolated from A. mellifera, is needed. PMID:22221381

Choe, Se-Eun; Nguyen, Thuy Thi-Dieu; Hyun, Bang-Hun; Noh, Jin-Hyeong; Lee, Hee-Soo; Lee, Chang-Hee; Kang, Seung-Won

2012-05-25

229

Unique RNA 2 sequences of two Brazilian isolates of Pepper ringspot virus, a tobravirus.  

PubMed

Pepper ringspot virus (PepRSV) is a tobravirus reported only in Brazil. Here, the sequences of the complete RNA 2 segments and the 3' end of the RNA 1 genomic regions of two new isolates from tomato plants were analyzed. The main ORF encodes the CP gene as other tobraviruses and termed ORF 1 of RNA 2. The second ORF was found only in one of the new isolates, although this gene was absent in the type isolate, CAM (collected in the 1960's). Interestingly, this ORF 2 gene did not show any nucleotide and amino acid sequence similarities with known 2b genes of tobraviruses, an essential gene of tobraviruses for nematodes-transmission. The 5'UTR sequence of RNA 2 segment of CAM isolate was previously reported showing two impaired direct repeats; however, the direct-repeats were absent in these new isolates. An additional ORF was predicted upstream of the CP gene. This putative protein possessed a transmembrane domain similar to the ORFN1 of RNA 2 of Tobacco rattle virus SYM isolate, although there was no sequence similarity. This is the first report on the diversity of the RNA 2 sequences of PepRSV. PMID:24756556

Batista, Adriana Ribeiro Silva; Nicolini, Cícero; Rodrigues, Kelly Barreto; Melo, Fernando Lucas; Vasques, Raquel Medeiros; de Macędo, Mônica Alves; Inoue-Nagata, Alice Kazuko; Nagata, Tatsuya

2014-08-01

230

Isolation and genetic characterization of Japanese encephalitis virus from equines in India  

PubMed Central

Japanese encephalitis (JE) is an important vector-borne viral disease of humans and horses in Asia. JE outbreaks occur regularly amongst humans in certain parts of India and sporadic cases occur among horses. In this study, JE seroprevalence and evidence of JE virus (JEV) infection among horses in Haryana (India) is described. Antibodies against JEV were detected in 67 out of 637 (10.5%) horses screened between 2006 and 2010. Two foals exhibiting neurological signs were positive for JEV RNA by RT-PCR; JEV was isolated from the serum of one of the foals collected on the second day of illness. This is the first report of JEV isolation from a horse in India. Furthermore, a pool of mosquitoes collected from the premises housing these foals was positive for JEV RNA by RT-PCR. Three structural genes, capsid (C), premembrane (prM), and envelope (E) of the isolated virus (JE/eq/India/H225/2009) spanning 2,500 nucleotides (from 134 to 2,633) were cloned and sequenced. BLAST results showed that these genes had a greater than 97% nucleotide sequence identity with different human JEV isolates from India. Phylogenetic analysis based on E- and C/prM genes indicated that the equine JEV isolate belonged to genotype III and was closely related to the Vellore group of JEV isolates from India. PMID:22705732

Singha, Harisankar; Singh, Birendra K.; Virmani, Nitin; Kumar, Sanjay; Singh, Raj K.

2012-01-01

231

Pathogenicity and transmission of reticuloendotheliosis virus isolated from endangered prairie chickens.  

PubMed

The pathogenicity and transmission of a field isolate of reticuloendotheliosis virus (REV) was studied using an experimental model in Japanese quail. Oncogenicity was also evaluated after inoculations in chickens and turkeys. The original REV (designated APC-566) was isolated from Attwater's prairie chickens (Tympanuchus cupido attwateri), an endangered wild avian species of the southern United States. The transmissibility of the REV isolate was studied in young naive Japanese quail in contact with experimentally infected quail. Vertical transmission was not detected by virus isolation and indirect immunofluorescence. Seroconversion was detected in few contact quails, suggesting horizontal transmission. The APC-566 isolate induced tumors beginning at 6 wk of age in quails infected as embryos. Most of the tumors detected in Japanese quail were lymphosarcomas, and 81% of these neoplasias contained CD3+ cells by immunoperoxidase. REV APC-566 was also oncogenic in chickens and turkeys infected at 1 day of age, with tumors appearing as early as 58 days after infection in chickens and at 13 wk of age in turkeys. This study was conducted in part as an attempt to understand the potential for pathogenicity and transmission of REV isolated from endangered avian species. PMID:17461264

Barbosa, Taylor; Zavala, Guillermo; Cheng, Sunny; Villegas, Pedro

2007-03-01

232

Identification of new isolates of Turnip mosaic virus that cluster with less common viral strains.  

PubMed

Turnip mosaic virus (TuMV) was found infecting cultivated brassicas and wild and cultivated ornamental Brassicaceae plants in different regions of Spain. Five new TuMV isolates, originating from different host plant species (Brassica cretica, Brassica juncea, Brassica napus, Eruca vesicaria subsp. sativa and Sisymbrium orientale), have been identified. The nucleotide sequences of the coat protein (CP) genes of the five isolates were determined. Phylogenetic analysis of the CP sequences showed that the five isolates grouped into two different clusters. The three isolates from the central region of Spain clustered with a previously reported Pisum sativum isolate from southeastern Spain, whereas the other two isolates from the eastern region clustered with two Italian and two Greek isolates. Both clusters were genetically distinct and belonged to the multi-lineage group OBR. The OBR group contains mainly TuMV isolates from hosts other than Brassica spp. and Raphanus sativus and mostly originating from Mediterranean countries. These new sequences provide further phylogenetic resolution of the OBR group. Although new TuMV isolates have been found in Spain, they were not associated with any serious disease outbreaks. PMID:17347771

Sánchez, F; Rodríguez-Mateos, M; Tourińo, A; Fresno, J; Gómez-Campo, C; Jenner, C E; Walsh, J A; Ponz, F

2007-01-01

233

Genetic and serological typing of European infectious haematopoietic necrosis virus (IHNV) isolates  

USGS Publications Warehouse

Infectious haematopoietic necrosis virus (IHNV) causes the lethal disease infectious haematopoietic necrosis (IHN) in juvenile salmon and trout. The nucleocapsid (N) protein gene and partial glycoprotein (G) gene (nucleotides 457 to 1061) of the European isolates IT-217A, FR-32/87, DE-DF 13/98 11621, DE-DF 4/99-8/99, AU-9695338 and RU-FR1 were sequenced and compared with IHNV isolates from the North American genogroups U, M and L. In phylogenetic studies the N gene of the Italian, French, German and Austrian isolates clustered in the M genogroup, though in a different subgroup than the isolates from the USA. Analyses of the partial G gene of these European isolates clustered them in the M genogroup close to the root while the Russian isolate clustered in the U genogroup. The European isolates together with US-WRAC and US-Col-80 were also tested in an enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies (MAbs) against the N protein. MAbs 136-1 and 136-3 reacted equally at all concentrations with the isolates tested, indicating that these antibodies identify a common epitope. MAb 34D3 separated the M and L genogroup isolates from the U genogroup isolate. MAb 1DW14D divided the European isolates into 2 groups. MAb 1DW14D reacted more strongly with DE-DF 13/98 11621 and RU-FR1 than with IT-217A, FR- 32/87, DE-DF 4/99-8/99 and AU-9695338. In the phylogenetic studies, the Italian, French, German and Austrian isolates clustered in the M genogroup, whereas in the serological studies using MAbs, the European M genogroup isolates could not be placed in the same specific group. These results indicate that genotypic and serotypic classification do not correlate. ?? 2009 Inter-Research.

Johansson, T.; Einer-Jensen, K.; Batts, W.; Ahrens, P.; Bjorkblom, C.; Kurath, G.; Bjorklund, H.; Lorenzen, N.

2009-01-01

234

Analysis of the biological and molecular variability of Watermelon mosaic virus isolates from Iran.  

PubMed

Watermelon mosaic virus (WMV) is one of the most important viruses that causes different symptoms in Cucurbitaceae. WMV is a potyvirus with a worldwide distribution, but occurs most commonly in temperate and Mediterranean regions. Cucurbit species grown in Yazd, Esfahan, West Azerbaijan, Hormozgan, and Kerman provinces were surveyed for the relative incidence of WMV in 2004-2005. A total of 757 symptomatic cucurbit and 31 weed species were collected and assayed for infection with WMV. Of 788 leaf samples from cucurbit and weed plants, 190 samples were positive by double antibody sandwich ELISA (DAS-ELISA) using specific polyclonal antibody. Among the weed species tested only colocynth (Citrullus colocynthis) was found to be infected with WMV. The coat protein (CP) gene from 18 representative isolates was PCR amplified, cloned, sequenced, and compared with the sequences available in GeneBank. Phylogenetic analysis using 778 nucleotide long sequences of the coat protein gene showed that these isolates fell into two; groups I and II. Only one isolates (KER.JI.1) was classified in the group II. This isolate had a wider host range and infected Nicotiana debneyii and Datura metel. None of the other 17 isolates could infect these two species. Members of group I were divided into three subgroups; A, B, and C. The subgroup I(B) appears to be a new subgroup comprising only of the Iranian isolates. Phylogenetic analysis based on 200 nucleotides coding for the N-terminal segment of the CP showed that all Iranian isolates except KER.JI.1 clustered with the previously reported WMV strains. All Iranian isolates had a DAG amino acid triplet which is involved in aphid transmissibility. This is the first report on sequence analysis of the nearly full-length CP cDNA clones of WMV isolates from Iran. PMID:18712590

Sharifi, Maraym; Massumi, Hossain; Heydarnejad, Jahangir; Hosseini Pour, Akbar; Shaabanian, Mehdi; Rahimian, H

2008-12-01

235

Antigenic and genotypical characterization of Newcastle disease viruses isolated in Taiwan between 1969 and 1996.  

PubMed

Three major epidemics of Newcastle disease (ND) occurred in Taiwan over the past three decades (in 1969, 1984, and 1995). In order to gain a better understanding of the relationships between past ND epizootics in Taiwan, 36 ND viruses (NDVs) isolated between 1969 and 1996 were characterized antigenically and genotypically. The antigenicity of these viruses was analysed by their ability to cause binding of mouse monoclonal antibodies (mAbs) to cell cultures infected with the isolate. Using a panel of 22 mAbs to divide NDVs into subgroups, a total of 18 binding patterns were revealed. The sequences covering the cleavage site of the fusion protein gene of these isolates were also determined. The results of the phylogenetic analysis placed 36 NDVs into I, II, VIb, VIIa, VIII and two novel genotypes (provisionally termed X and VIh). The 1969 velogenic isolates were of genotypes X and VIh; the 1984-1985 velogenic isolates were genotyped VIb, VIh, VIIa, and X; while the 1995-1996 velogenic isolates were genotyped VIIa or VIII. Some 1969 and 1984 velogenic isolates were of the same mAbs binding pattern and genotype, and the mAbs binding patterns of the 1995-1996 isolates have not been seen before. It is concluded that velogenic NDVs of different genotype and antigenic type have co-circulated in Taiwan at least since 1969. Also there were epizootiological links between strains isolated in 1969 and 1984, whereas the 1995-1996 epidemic was caused by new antigenic variants. PMID:15530736

Tsai, Hsiang-Jung; Chang, Kuo-Hui; Tseng, Chun-Hsien; Frost, Karen M; Manvell, Ruth J; Alexander, Dennis J

2004-11-30

236

Surveillance and characterization of Newcastle disease viruses isolated from northern pintail (Anas acuta) in Japan during 2006-09.  

PubMed

A total of 38 Newcastle disease virus (NDV) isolates were obtained from 6060 fecal samples from northern pintail (Anas acuta) ducks collected in the Tohoku district in Japan during 2006-09. One isolate from each sampling location and date was selected for a total of 38 isolates, then 15 of these were characterized for their pathogenicity by mean death time of minimum lethal dose (MDT/MLD) using chicken embryos and by plaque formation on chicken embryo fibroblasts. Furthermore, nine isolates were randomly selected from these 15 isolates, and the fusion protein genes were sequenced to characterize amino acid sequences around the cleavage site. All 15 were confirmed to be nonvirulent by MDT/MLD test, and nine isolates were also confirmed as nonvirulent by the cleavage site of the fusion protein 112G/E-K/R-Q-G/E-R*L117 that was specific for nonvirulent NDVs. The characteristics of nine isolates identified by phylogenic analysis of the fusion protein gene indicated that the isolates belong to genotype I or II. In addition, we also isolated 68 avian influenza viruses and 28 other hemagglutinating viruses. Our data indicate that northern pintails are subclinically infected by, perpetuate, and distribute NDV along with different subtypes of avian influenza viruses and other hemagglutinating viruses during their migrations across vast areas over the Northern Hemisphere to Japan. PMID:21793438

Ruenphet, Sakchai; Jahangir, Alam; Shoham, Dany; Morikawa, Kae; Miyoshi, Yuki; Hanawa, Eiko; Okamura, Masashi; Nakamura, Masayuki; Takehara, Kazuaki

2011-06-01

237

Isolation and titration of dengue viruses by the mosquito inoculation technique.  

PubMed

Mosquito inoculation is a highly sensitive technique for isolation and titration of dengue virus (DENV) from sera, human tissues, wild animals, or mosquitoes. It has been under utilized since it was described 40 years ago because most dengue laboratories do not have access to an insectary to rear mosquitoes. This technique requires good eye-hand coordination while doing manipulation under a stereoscopic microscope, and extensive practice is needed to become proficient at inoculating mosquitoes. Following inoculation, mosquitoes are held for 10 days to allow dengue virus to replicate and disseminate to tissues throughout the mosquitoes. They are then harvested and examined for the presence of viral antigens in head tissue by either immunofluorescence assay (IFA) or PCR (polymerase chain reaction). The mosquito infectious dose 50 (MID50) is calculated using the method of Reed and Muench to quantitate the virus. This method can be used for other arboviruses as well as for dengue. PMID:24696328

Choy, Milly M; Gubler, Duane J

2014-01-01

238

Complete genome sequence of invertebrate iridescent virus 22 isolated from a blackfly larva  

PubMed Central

Members of the family Iridoviridae are animal viruses that infect only invertebrates and poikilothermic vertebrates. Invertebrate iridescent virus 22 (IIV-22) was originally isolated from the larva of a blackfly (Simulium sp., order Diptera) found in the Ystwyth river, near Aberystwyth, Wales, UK. IIV-22 virions are icosahedral, with a diameter of about 130 nm and contain a dsDNA genome that is 197.7 kb in length, has a G+C content of 28.05 mol% and contains 167 coding sequences. Here, we describe the complete genome sequence of this virus and its annotation. This is the fourth genome sequence of an invertebrate iridovirus to be reported. PMID:23804567

Piegu, Benoit; Guizard, Sebastien; Spears, Tatsinda; Cruaud, Corinne; Couloux, Arnault; Bideshi, Dennis K.; Federici, Brian A.

2013-01-01

239

Molecular characterization of avian-like H1N1 swine influenza a viruses isolated in Eastern China, 2011.  

PubMed

Currently, three predominant subtypes of influenza virus are prevalent in pig populations worldwide: H1N1, H3N2, and H1N2. European avian-like H1N1 viruses, which were initially detected in European pig populations in 1979, have been circulating in pigs in eastern China since 2007. In this study, six influenza A viruses were isolated from 60 swine lung samples collected from January to April 2011 in eastern China. Based on whole genome sequencing, molecular characteristics of two isolates were determined. Phylogenetic analysis showed the eight genes of the two isolates were closely related to those of the avian-like H1N1 viruses circulating in pig populations, especially similar to those found in China. Four potential glycosylation sites were observed at positions 13, 26, 198, 277 in the HA1 proteins of the two isolates. Due to the presence of a stop codon at codon 12, the isolates contained truncated PB1-F2 proteins. In this study, the isolates contained 591Q, 627E and 701N in the polymerase subunit PB2, which had been shown to be determinants of virulence and host adaptation. The isolates also had a D rather than E at position 92 of the NS1, a marker of mammalian adaptation. Both isolates contained the GPKV motif at the PDZ ligand domain of the 3' end of the NS1, a characteristic marker of the European avian-like swine viruses since about 1999, which is distinct from those of avian, human and classical swine viruses. The M2 proteins of the isolates have the mutation (S31N), a characteristic marker of the European avian-like swine viruses since about 1987, which may confer resistance to amantadine and rimantadine antivirals. Our findings further emphasize the importance of surveillance on the genetic diversity of influenza A viruses in pigs, and raise more concerns about the occurrence of cross-species transmission events. PMID:23055004

Qi, Xian; Pan, Yuning; Qin, Yuanfang; Zu, Rongqiang; Tang, Fengyang; Zhou, Minghao; Wang, Hua; Song, Yongchun

2012-10-01

240

Sequence conservation in field and experimental isolates of Borna disease virus.  

PubMed Central

Coding and noncoding sequences were analyzed from field and experimental isolates of Borna disease virus. For a 24-kDa protein, maximum divergence was 1.5% at the predicted amino acid level and 3.1% at the nucleotide level. For a 40-kDa protein, maximum divergence was 1.1% at the predicted amino acid level and 4.1% at the nucleotide level. The highest variability in sequence (10%) was found in a 40-nucleotide stretch of genomic RNA between coding sequences for the 40- and 24-kDa proteins. The degree of sequence conservation in these isolates, passaged in various host species in vivo and in vitro over a period of 64 years, is unusual for negative-strand RNA viruses. PMID:8254777

Schneider, P A; Briese, T; Zimmermann, W; Ludwig, H; Lipkin, W I

1994-01-01

241

Comparative study of influenza virus H2 (Asian) hemagglutinins isolated from human and avian sources.  

PubMed

The hemagglutinin of an influenza virus isolated from a wild duck (Pintail, Anas acuta) in the USSR in 1976 had been found to be antigenically indistinguishable from the hemagglutinin of H2N2 viruses of human origin isolated in 1957. The hemagglutinins from viral preparations of the A/Anas acuta/Primorie/695/76 (H2Nav2) and A/Singapore/1/57 (H2N2) strains were purified by SDS gel chromatography as the subunits HA1 and HA2. Comparison of amino acid compositions and peptide maps of tryptic peptides containing [14C]-carboxymethylcysteine showed a striking degree of similarity between the H2 hemagglutinins. PMID:7193663

Bucher, D J; Kharitonenkov, I G; Lvov, D K; Pysina, T V; Lee, H M

1980-01-01

242

Genetic Diversity of the Coat Protein of Olive Mild Mosaic Virus (OMMV) and Tobacco Necrosis Virus D (TNV-D) Isolates and Its Structural Implications  

PubMed Central

The genetic variability among 13 isolates of Olive mild mosaic virus (OMMV) and of 11 isolates of Tobacco necrosis virus D (TNV-D) recovered from Olea europaea L. samples from various sites in Portugal, was assessed through the analysis of the coat protein (CP) gene sequences. This gene was amplified through reverse transcriptase polymerase chain reaction (RT-PCR), cloned, and 5 clone sequences of each virus isolate, were analysed and compared, including sequences from OMMV and TNV-D isolates originally recovered from different hosts and countries and available in the GenBank, totalling 131 sequences. The encoded CP sequences consisted of 269 amino acids (aa) in OMMV and 268 in TNV-D. Comparison of the CP genomic and amino acid sequences of the isolates showed a very low variability among OMMV isolates, 0.005 and 0.007, respectively, as well as among TNV-D isolates, 0.006 and 0.008. The maximum nucleotide distances of OMMV and TNV-D sequences within isolates were also low, 0.013 and 0.031, respectively, and close to that found between isolates, 0.018 and 0.034, respectively. In some cases, less variability was found in clone sequences between isolates than in clone sequences within isolates, as also shown through phylogenetic analysis. CP aa sequence identities among OMMV and TNV-D isolates ranged from 84.3% to 85.8%. Comparison between the CP genomic sequences of the two viruses, showed a relatively low variability, 0.199, and a maximum nucleotide distance between isolates of 0.411. Analysis of comparative models of OMMV and TNV-D CPs, showed that naturally occurring substitutions in their respective sequences do not seem to cause significant alterations in the virion structure. This is consistent with a high selective pressure to preserve the structure of viral capsid proteins. PMID:25350108

Varanda, Carla M. R.; Machado, Marco; Martel, Paulo; Nolasco, Gustavo; Clara, Maria I. E.; Félix, Maria R.

2014-01-01

243

Identification of syncytial mutations in a clinical isolate of herpes simplex virus 2  

SciTech Connect

Small polykaryocytes resulting from cell fusion are found in herpes simplex virus (HSV) lesions in patients, but their significance for viral spread and pathogenesis is unclear. Although syncytial variants causing extensive fusion in tissue culture can be readily isolated from laboratory strains, they are rarely found in clinical isolates, suggesting that extensive cell fusion may be deleterious in vivo. Syncytial mutations have previously been identified for several laboratory strains, but not for clinical isolates of HSV type 2. To address this deficiency, we studied a recent syncytial clinical isolate, finding it to be a mixture of two syncytial and one nonsyncytial strain. The two syncytial strains have novel mutations in glycoprotein B, and in vitro cell fusion assays confirmed that they are responsible for syncytium formation. This panel of clinical strains may be ideal for examining the effect of increased cell fusion on pathogenesis.

Muggeridge, Martin I. [Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, LA 71130 (United States) and Center for Molecular and Tumor Virology, Louisiana State University Health Sciences Center, Shreveport, LA 71130 (United States) and Feist-Weiller Cancer Center, Louisiana State University Health Sciences Center, Shreveport, LA 71130 (United States)]. E-mail: mmugge@lsuhsc.edu; Grantham, Michael L. [Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, LA 71130 (United States); Center for Molecular and Tumor Virology, Louisiana State University Health Sciences Center, Shreveport, LA 71130 (United States); Johnson, F. Brent [Department of Microbiology and Molecular Biology, Brigham Young University, Provo, UT 84602 (United States)

2004-10-25

244

Comparative transmission of two cucumber mosaic virus isolates by two color morphs of Acyrthosiphon pisum (Harris).  

PubMed

 Cucumber mosaic virus (CMV) is one of the most important legume-infecting viruses, which is transmitted effectively by pea aphid Acyrthosiphon pisum (Harris) (Hem: Aphididae). Transmission efficiency of two CMV isolates (As and Kh from cowpea and bean hosts, resp.) by red and green color morphs of pea aphid were evaluated on bean plants. Triple-antibody sandwich ELISA (TAS-ELISA) using CMV-specific monoclonal antibodies revealed that both CMV isolates belonged to the serotype II. Bean plants inoculated by viruliferous aphids were assayed by double-antibody sandwich ELISA (DAS-ELISA) at 16 days post inoculation (dpi). The results showed that the transmission rate of CMV-As by the red morph was significantly higher than by the green morph, resulting in significantly higher transmission rate of the virus (As + Kh) by the red morph than by the green morph, with p? 0.1. Similarly, the efficiency of CMV transmission by the red morph of A. pisum was higher than the efficiency of transmission by the green morph. The higher transmission rate and efficiency of CMV by red pea aphid would be important in the epidemiology. Based on these results, we hypothesize that the transmission efficiency of CMV is affected more by the difference in transmission determinants of A. pisum color morphs than by the sequence of virus coat protein determinants. Keywords: Aphididae; Bromoviridae; color polymorphism; transmission efficiency. PMID:22720705

Tahmasebi, A; Dizadji, A; Farhoudi, F; Allahyari, H; Koohi-Habibi, M

2012-01-01

245

Sea bass Dicentrarchus labrax nervous necrosis virus isolates with distinct pathogenicity to sea bass larvae.  

PubMed

Reproduction of nodavirus disease was performed by experimental infection of sea bass eggs during fertilization or at larval stage 4 with 2 genetically distinguishable nodavirus strains (Sb1 and Sb2) isolated from sea bass collected along the Atlantic and Mediterranean French coast. The pathogenicity of the virus strains was assigned after detection of the virus by ELISA and immunohistochemistry (IHC). The Atlantic (Sb1) strain was more pathogenic than the Mediterranean (Sb2) strain during the fertilization step whilst both strains were pathogenic following experimental exposure of 4 d old larvae. Virus lesions developed in the brain 4 to 6 d following experimental exposure. Experimental ELISA proved very sensitive for detecting the nodavirus in Sb1 or Sb2 experimentally infected larvae, as well as in naturally infected sea bass larvae collected in French hatcheries or in barramundi larvae reared in the Pacific area. The development of an ELISA specific for the 2 nodavirus strains isolated from the sea bass should be useful for the detection of the virus, in addition to other techniques recommended by the Office International des Epizooties (OIE). PMID:11411641

Breuil, G; Mouchel, O; Fauvel, C; Pepin, J F

2001-05-01

246

Genetic Diversity of a Natural Population of Apple stem pitting virus Isolated from Apple in Korea  

PubMed Central

Apple stem pitting virus (ASPV), of the Foveavirus genus in the family Betaflexiviridae, is one of the most common viruses of apple and pear trees. To examine variability of the coat protein (CP) gene from ASPV, eight isolates originating from 251 apple trees, which were collected from 22 apple orchards located in intensive apple growing areas of the North Gyeongsang and North Jeolla Provinces in Korea, were sequenced and compared. The nucleotide sequence identity of the CP gene of eight ASPV isolates ranged from 77.0 to 97.0%, while the amino acid sequence identity ranged from 87.7 to 98.5%. The N-terminal region of the viral CP gene was highly variable, whereas the C-terminal region was conserved. Genetic algorithm recombination detection (GARD) and single breakpoint recombination (SBP) analyses identified base substitutions between eight ASPV isolates at positions 54 and 57 and position 771, respectively. GABranch analysis was used to determine whether the eight isolates evolved due to positive selection. All values in the GABranch analysis showed a ratio of substitution rates at non-synonymous and synonymous sites (dNS/dS) below 1, suggestive of strong negative selection forces during ASPV CP history. Although negative selection dominated CP evolution in the eight ASPV isolates, SLAC and FEL tests identified four possible positive selection sites at codons 10, 22, 102, and 158. This is the first study of the ASPV genome in Korea.

Yoon, Ju Yeon; Joa, Jae Ho; Choi, Kyung San; Do, Ki Seck; Lim, Han Cheol; Chung, Bong Nam

2014-01-01

247

Isolation and partial characterisation of a new strain of Ebola virus  

Microsoft Academic Search

We have isolated a new strain of Ebola virus from a non-fatal human case infected during the autopsy of a wild chimpanzee in the Cote-d'Ivoire. The wild troop to which this animal belonged has been decimated by outbreaks of haemorrhagic syndromes. This is the first time that a human infection has been connected to naturally-infected monkeys in Africa. Data from

B. Le Guenno; P Formentry; M. Wyers; P. Gounon; F. Walker; C. Boesch

1995-01-01

248

The Isolation and some Properties of a Virus-Inhibiting Protein from Phytolacca esculenta  

Microsoft Academic Search

SUMMARY: An inhibitor of plant viruses can be isolated from the sap of Phytolacca esculenta by differential precipitation with ethanol followed by adsorption on celite and elution with 10% NaC1. Purified preparations contain 141-15% nitrogen and 8-12 yo carbohydrate and the inhibitor is probably a glycoprotein. Denaturation leads to loss of inhibiting power. The protein, unless denatured, !s unaffected by

B. Kassanis; A. Kleczkowski

1948-01-01

249

Identification of 5' capped structure and 3' terminal sequence of hepatitis E virus isolated from Morocco  

Microsoft Academic Search

AIM: To examine 5' and 3' terminal sequences of hepatitis E virus (HEV) isolated from Morocco, to confirm 5' methylated cap structure of the genome, and to investigate whether the 3' UTR can be used to distinguish HEV genotypes instead of HEV complete genome sequence. METHODS: RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE) was employed to obtain the 5'

Guo-Bing Chen; Ji-Hong Meng

2004-01-01

250

Molecular characterization of the first Australian isolate of Japanese encephalitis virus, the FU strain  

Microsoft Academic Search

The complete genomic and predicted amino acid sequence of the Japanese encephalitis virus (JEV) FU strain, a human isolate recovered from the first outbreak of Japanese encephalitis in Australian territory, was determined. Comparison of the FU genome with 15 fully sequenced JEV genomes revealed high levels of sequence identity, ranging from 88-7% (GP78) to 89-7% (K94P05) for nucleotides and 96-8%

David T. Williams; Lin-Fa Wang; Peter W. Daniels; John S. Mackenzie

2000-01-01

251

Characterization of Tissue Culture-induced Heterogeneity in DNAs of Independent Isolates of JC Virus  

Microsoft Academic Search

SUMMARY After several serial passages at low multiplicities of infection in primary human foetal glial cells at 37 °C, the DNA of prototype (MAD-I) JC virus and that of MAD-2 and MAD-3 are typically heterogeneous in size, but DNAs of MAD-4 and MAD-6 are relatively homogeneous. A similar dichotomy was observed in the DNAs of six isolates propagated more recently

JONATHAN D. MARTIN; BILLIE L. PADGETT; DUARD L. WALKER

1983-01-01

252

Antigenic variants of rabies virus in isolates from eastern, central and northern Canada.  

PubMed Central

Street rabies virus isolated from 51 specimens from Ontario, Quebec, Manitoba and the Northwest Territories have been typed by a panel of 36 antinucleocapsid monoclonal antibodies. Three main groups were found. The first group comprised those terrestrial mammals originating in Ontario, Quebec and the Northwest Territories. The second group was found in terrestrial mammals from Manitoba. The third heterogenous group was made up of bats from Ontario. PMID:3893660

Webster, W A; Casey, G A; Charlton, K M; Wiktor, T J

1985-01-01

253

Complete nucleotide sequence of a Japanese isolate of Chrysanthemum virus B (genus Carlavirus )  

Microsoft Academic Search

Summary  The complete nucleotide sequence of a Chrysanthemum virus B isolate from Japan (CVB-S) has been determined. The genomic RNA of CVB-S is 8,990 nucleotides long, excluding the poly(A)\\u000a tail and, like that of other carlaviruses, contains six open reading frames (ORFs). Multiple alignment and phylogenetic analyses\\u000a indicated that the phylogenetic relationship among members of the genus Carlavirus is very diverse,

A. Ohkawa; M. Yamada; H. Sayama; N. Sugiyama; S. Okuda; T. Natsuaki

2007-01-01

254

Phylogenetic study on the 5'-untranslated region of bovine viral diarrhoea virus isolates from Iran.  

PubMed

Bovine viral diarrhoea virus is a pathogen of bovids associated with reproduction system, causing in infected animals a range of ailments, from abortion to congenital defects. In this article, the nucleotide structure of the 5'-untranslated region (5-UTR) from 7 Iranian bovine diarrhoea virus (BVDV) isolates was characterized and subjected to comparative analysis against a panel of BVDV isolates from different sources. To this end, a 288 bp-long stretch of the internal ribosome entry site was amplified by RT-PCR. The PCR products subsequently cloned into PTZ57T vector and sequenced using T7 promoter primers. This resulted in detection of 3 new point mutations G ? A and G ? T in 2 isolates. When these findings were phylogenetically assessed, all the examined Iranian isolates were deemed to belong to the type1 of BVDV. Besides, 2 subtypes were identified among these isolates. In group A, a high level of similarity (99.2%) between Iranian isolates with a cytopathic Australian strain of BVDV-1c was detected; while in group B, the 4 Iranian isolates proved to be very similar to NADL-like BVDV-1a strains. We believe that the surprisingly high level of similarity between group A Iranian isolates and their corresponding Australian strain is likely to be an indication of a shared common ancestor. If correct, the most likely explanation of this observation is the introduction of such strains from Australia to Iran, possibly through exportation of infected live animals or animal productions (e.g. semen and meat) at some points in the past. Nevertheless, this hypothesis remains to be proved as further epidemiological work at genomic level is required to understand population of BVDV in Iran. PMID:25273964

Esmaelizad, Majid; Kargar-Moakhar, Rohani

2014-09-30

255

Receptor Specificity and Transmission of H2N2 Subtype Viruses Isolated from the Pandemic of 1957  

E-print Network

Influenza viruses of the H2N2 subtype have not circulated among humans in over 40 years. The occasional isolation of avian H2 strains from swine and avian species coupled with waning population immunity to H2 hemagglutinin ...

Pappas, Claudia

256

Infectivity analysis of a blackgram isolate of Mungbean yellow mosaic virus and genetic assortment with MYMIV in selective hosts.  

PubMed

Yellow mosaic disease in grain legumes in Indian subcontinent is caused by two important virus species viz. Mungbean yellow mosaic virus (MYMV) and Mungbean yellow mosaic India virus (MYMIV), belonging to the genus Begomovirus of the family Geminiviridae. The genomic components of a begomovirus causing yellow mosaic disease in blackgram in southern India were cloned and sequenced. Nucleotide sequence comparison of DNA A component shows the virus isolate to be a variant of Mungbean yellow mosaic virus:-(MYMV-[IN:Vam:05]). However, DNA B component of the present virus isolate has greater similarity (92%) to Mungbean yellow mosaic India virus. Agroinoculations of the viral clones produced typical yellow mosaic symptoms in blackgram and mungbean, severe leaf curl and stunting in French bean, similar to blackgram isolate of MYMIV. Blackgram isolates of both the virus species were only mildly infectious on cowpea, produced atypical leaf curl symptoms and not yellow or golden mosaic. In agroinoculations done by exchanging genomic components, symptom expression was seen only in French bean. In cowpea, blackgram and mungbean there was no visible symptoms though viral DNA could be detected by PCR. PMID:21380756

Haq, Q M I; Rouhibakhsh, A; Ali, Arif; Malathi, V G

2011-06-01

257

Complete genome sequence of a tobacco isolate of the tobacco vein banding mosaic virus strain prevailing in China  

Microsoft Academic Search

Tobacco vein banding mosaic virus (TVBMV) is a species of the largest plant virus genus Potyvirus. Its incidence has been increasing in Chinese tobacco-growing area. TVBMV isolates can be clustered into three genetic groups\\u000a that are corresponding with their geographical origin. We have reported the complete genomic sequence of TVBMV isolate YND\\u000a with unique NIb\\/CP cleavage site. Here, we determined

H.-Y. Wang; T.-S. Zhu; T.-T. Cui; S.-S. Hou; X. Yin; Xiang-Dong Li; L.-P. Lei; X.-P. Zhu

2010-01-01

258

CHARACTERIZATION OF H5N1 INFLUENZA VIRUSES ISOLATED FROM MIGRATORY BIRDS IN QINGHAI PROVINCE OF CHINA IN 2006  

Microsoft Academic Search

SUMMARY. Avian influenza H5N1 viruses pose a significant threat to human health because of their ability to infect humans directly. In the paper, three highly pathogenic H5N1 influenza viruses were isolated from three species of migratory birds in Qinghai Province of China in 2006. The analysis of the genome sequences indicated that the three isolates shared high homology with each

Fumin Lei; AF Shuang Tang; A Xiaowei Zhang; Zhong Zhang; A Shengliang Chen; D Dehai Zhang; D Baoping Yan; Zuohua Yin; Shengliang Chen; Sandan Li; Dehai Zhang; Baoping Yan; Tianxian Li

2007-01-01

259

Genetic characterization of highly pathogenic H5N1 avian influenza viruses isolated from poultry farms in Egypt  

Microsoft Academic Search

Twenty-four avian influenza viruses were collected from poultry farms in three different governorates in Egypt during the\\u000a years 2006–2009 and genetically characterized. All the isolates were confirmed to be type A and subtype H5 influenza virus\\u000a by chromatographic strip test and hemagglutination inhibition assay. The sequence and phylogenetic data revealed that all\\u000a Egyptian isolates cluster together and belong to subclade

Abd Elfattah H. Eladl; Kamel I. Abou El-Azm; Abd Elshakour N. Ismail; Ahmed Ali; Yehia M. Saif; Chang-Won Lee

260

Surveillance for avirulent Newcastle disease viruses in domestic ducks (Anasplatyrhynchos and Cairina moschata) at live bird markets in Eastern China and characterization of the viruses isolated  

Microsoft Academic Search

We isolated and identified 201 Newcastle disease viruses (NDVs) from domestic ducks in a 5-year surveillance study at live bird markets in Eastern China. Seventy-three of these isolates were characterized biologically and genetically. Fusion protein (F) genes of these isolates were amplified by reverse transcription-polymerase chain reaction and sequenced. Intracerebral pathogenicity index tests in 1-day-old specific-pathogen-free chickens and the mean

Xiaowen Liu; Xiaoquan Wang; Shuang Wu; Shunlin Hu; Yi Peng; Feng Xue; Xiufan Liu

2009-01-01

261

[Phylogenetic analysis of the nucleotide sequences of Chatanga virus strains, the new representative of California encephalitis serocomplex, isolated in different regions of the Russian Federation].  

PubMed

The complete nucleotide sequences of the S segment and the fragments of M and L segments (on 900 and 410 nucleotides, respectively) were determined in 14 California encephalitis serocomplex (CES) strains isolated from different regions of the Russian Federation. Phylogenetic analysis by the sequences of genomic S, M, and L segments indicated that all Russian strains were an individual independent CES virus. The new virus is named Chatanga for the place of isolation of one of the strains. Two genetic groups of Chatanga virus have been identified. Snowshoe hare (Lepus americanus) virus and La Crosse virus are closest to Chatanga virus among CES viruses. PMID:19172903

Lavrent'ev, M V; Prilipov, A G; L'vov, S D; L'vov, D K

2008-01-01

262

A highly divergent Encephalomyocarditis virus isolated from nonhuman primates in Singapore  

PubMed Central

Background In 2001 and 2002, fatal myocarditis resulted in the sudden deaths of four, two adult and two juvenile, orang utans out of a cohort of 26 in the Singapore Zoological Gardens. Methods Of the four orang utans that underwent post-mortem examination, virus isolation was performed from the tissue homogenates of the heart and lung obtained from the two juvenile orang utans in Vero cell cultures. The tissue culture fluid was examined using electron microscopy. Reverse transcription and polymerase chain reaction with Encephalomyocarditis virus (EMCV)-specific primers targeting the gene regions of VP3/VP1 and 3D polymerase (3Dpol) confirmed the virus genus and species. The two EMCV isolates were sequenced and phylogenetic analyses of the virus genes performed. Serological testing on other animal species in the Singapore Zoological Gardens was also conducted. Results Electron microscopy of the two EMCV isolates, designated Sing-M100-02 and Sing-M105-02, revealed spherical viral particles of about 20 to 30 nm, consistent with the size and morphology of members belonging to the family Picornaviridae. In addition, infected-Vero cells showed positive immunoflorescence staining with antiserum to EMCV. Sequencing of the viral genome showed that the two EMCV isolates were 99.9% identical at the nucleotide level, indicating a similar source of origin. When compared with existing EMCV sequences in the VP1 and 3Dpol gene regions, the nucleotide divergence were at a maximum of 38.8% and 23.6% respectively, while the amino acid divergence were at a maximum of 33.9% and 11.3% respectively. Phylogenetic analyses of VP1 and 3Dpol genes further grouped the Sing-M100-02 and Sing-M105-02 isolates to themselves, away from existing EMCV lineages. This strongly suggested that Sing-M100-02 and Sing-M105-02 isolates are highly divergent variants of EMCV. Apart from the two deceased orang utans, a serological survey conducted among other zoo animals showed that a number of other animal species had neutralizing antibodies to Sing-M105-02 isolate, indicating that the EMCV variant has a relatively wide host range. Conclusions The etiological agent responsible for the fatal myocarditis cases among two of the four orang utans in the Singapore Zoological Gardens was a highly divergent variant of EMCV. This is the first report of an EMCV infection in Singapore and South East Asia. PMID:23914943

2013-01-01

263

Surveillance for avirulent Newcastle disease viruses in domestic ducks (Anas platyrhynchos and Cairina moschata) at live bird markets in Eastern China and characterization of the viruses isolated.  

PubMed

We isolated and identified 201 Newcastle disease viruses (NDVs) from domestic ducks in a 5-year surveillance study at live bird markets in Eastern China. Seventy-three of these isolates were characterized biologically and genetically. Fusion protein (F) genes of these isolates were amplified by reverse transcription-polymerase chain reaction and sequenced. Intracerebral pathogenicity index tests in 1-day-old specific-pathogen-free chickens and the mean death time of embryonated fowl eggs in addition to the cleavage site analysis of the F-protein precursor for these viruses showed that they were all avirulent NDVs. Phylogenetic analysis based on partial sequences of the F gene showed that 30 isolates clustered into the class I clade and the other 43 isolates clustered into genotype I of class II, but diverged from the vaccine virus Queensland V4, which is extensively used in China. Most class I viruses (18/30) formed a separate branch closest to the Hong Kong live bird market strains that have been recently designated as genotype 3, while the rest (12/30) were closely related to some European viruses within genotype 2. All of the 43 class II genotype I viruses diverged from viruses originally assigned to genotype Ia and formed a separate sublineage designated as Ib with water bird isolates from the Far East, suggesting the possible transmission between the wild and domestic waterfowl. The results in the present study clearly showed that the domestic duck population carries avirulent NDVs with genetic divergence regularly and may act as one of the important reservoirs. PMID:19937525

Liu, Xiaowen; Wang, Xiaoquan; Wu, Shuang; Hu, Shunlin; Peng, Yi; Xue, Feng; Liu, Xiufan

2009-10-01

264

The consequence of passive administration of an anti-human immunodeficiency virus type 1 neutralizing monoclonal antibody before challenge of chimpanzees with a primary virus isolate.  

PubMed Central

The anti-gp41 virus neutralizing monoclonal antibody 2F5 was infused into chimpanzees, which were then given an intravenous challenge with a primary human immunodeficiency virus type I (HIV-1) isolate. In two control animals, the infection was established immediately, as evidenced by positive cell-associated DNA PCR and serum RNA PCR tests within 1 week, seroconversion by 4 weeks, and development of lymphadenopathy in this acute phase. Serum RNA PCR tests were negative in one of the two antibody-infused animals until week 8 and in the other antibody-infused animal until week 12; both animals seroconverted at week 14. The peak of measurable virus-specific serum RNA was delayed until week 16 in one antibody-infused animal. Virus-specific RNA in the other animal did not reach levels comparable to those in the other animals through 1 year of follow-up studies. Virus was isolated from the week 16 blood sample from one infused animal. Virus was not isolated from peripheral blood of the second animal but was isolated from lymph node cells taken at week 36. The infection of untreated chimpanzees with this primary isolate appears robust. Use of this isolate should widen the scope of possible experiments in the chimpanzee model. This antibody infusion study indicates that neutralizing antibody, when present at the time of challenge, affects the timing and level of infection and remains influential after it can no longer be detected in the peripheral circulation. It is possible that preexisting, neutralizing antibodies (passively administered or actively elicited) affect the course of acute-phase virus replication in humans. It remains to be established whether these immunologically mediated early effects will influence the course of HIV-1 disease. PMID:8794312

Conley, A J; Kessler JA, I I; Boots, L J; McKenna, P M; Schleif, W A; Emini, E A; Mark, G E; Katinger, H; Cobb, E K; Lunceford, S M; Rouse, S R; Murthy, K K

1996-01-01

265

Complete nucleotide sequence of a South African isolate of Grapevine fanleaf virus.  

PubMed

The complete sequences of RNA1 and RNA2 have been determined for a South African isolate of Grapevine fanleaf virus (GFLV-SAPCS3). The two RNAs are, respectively, 7,342 and 3,817 nucleotides in length, excluding the poly(A) tails. RNA1 has a large open reading frame (ORF) of 6,852 nucleotides and a 5'-UTR and a 3'-UTR of 243 and 244 nucleotides, respectively. RNA2 encodes for an ORF of 3,330 nucleotides and has the highest nucleotide identity (90.4 %) with GFLV-F13. The full length nucleotide sequence of GFLV-SAPCS3 RNA1 had the highest nucleotide identity (86.5 %) to the French isolate GFLV-F13. The 5'- and 3'-UTRs of GFLV-SAPCS3 RNA2 are 272 nucleotides and 212 nucleotides (nt) in length, respectively. The GFLV-SAPCS3 RNA2 5'-UTR is 32-53 nt longer compared to other GFLV isolates. The GFLV-SAPCS3 RNA2 5'-UTR is also more closely related to GFLV-GHu and Arabis mosaic virus (ArMV) isolates than to other GFLV isolates. Putative intra- and interspecies recombination events between GFLV and ArMV isolates involving GFLV-SAPCS3 RNA1 and RNA2 were investigated. Recombination analysis software has indicated that the GFLV-SAPCS3 5'-UTR might have evolved from a recombinational event between GFLV-F13-type and ArMV-Ta-type isolate. PMID:22669541

Lamprecht, Renate L; Maree, Hans J; Stephan, Dirk; Burger, Johan T

2012-10-01

266

Diversity of caprine arthritis-encephalitis virus promoters isolated from goat milk and passaged in vitro.  

PubMed

Transcriptional regulation in retroviruses resides in the U3 region of the proviral long terminal repeat (LTR). Transcription binding sites (TBS) in the U3 region of proviral sequences derived from the milk of 17 goats infected with caprine arthritis-encephalitis virus (CAEV) were analysed by nested PCR and sequencing. U3 sequences shared a high degree of homology (86-99%) and were closely related to isolates previously ascribed to small ruminant lentivirus subtype B1. Multiple putative AP-1, AP-4, Ets-1, Stat-1 and TATA binding protein (TBP) sites were highly conserved (>85% of isolates), as were single AML(vis), GAS, IRF-1, NFAT and TAS sites. A 10 nucleotide insertion of undetermined relevance was identified in the U3 region of two isolates. To study the stability of TBS within the CAEV U3 region through in vitro passage, milk-derived isolates of CAEV from three infected dams were cultured in goat synovial membrane (GSM) cells; in one isolate the viral U3 region was completely stable during in vitro passage, in a second isolate the viral U3 region accumulated multiple deletions, single nucleotide polymorphisms and insertions, while a third isolate had an intermediate degree of promoter stability. Promoter mutations arising during in vitro passage did not affect most of the conserved putative TBS identified in CAEV. PMID:23183018

Gomez-Lucia, Esperanza; Rowe, Joan; Collar, Carol; Murphy, Brian

2013-06-01

267

Molecular analysis of soybean dwarf virus isolates in the eastern United States confirms the presence of both D and Y strains and provides evidence of mixed  

E-print Network

Molecular analysis of soybean dwarf virus isolates in the eastern United States confirms for revision 24 October 2010 Accepted 4 January 2011 Available online 21 January 2011 Keywords: Soybean dwarf virus Recombination Mixed infections Emerging plant virus Soybean dwarf virus (SbDV), first identified

Graves, Michael V.

268

A minute virus of canines (MVC: canine bocavirus) isolated from an elderly dog with severe gastroenteritis, and phylogenetic analysis of MVC strains  

Microsoft Academic Search

Two of the three adult dogs kept in a family developed severe gastroenteritis. From the feces of one of the affected dogs a minute virus of canines (MVC) was detected by PCR and virus isolation. That this virus had recently infected the dogs was indicated by high anti-MVC antibody titers of their sera. No other virus commonly associated with canine

T. Ohshima; K. Kawakami; T. Abe; M. Mochizuki

2010-01-01

269

The isolation of salmonellae, Newcastle disease virus and other infectious agents from quarantined imported birds in Canada.  

PubMed Central

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. PMID:7039785

Rigby, C E; Pettit, J R; Papp-Vid, G; Spencer, J L; Willis, N G

1981-01-01

270

The complete genome sequence of a south Indian isolate of Rice tungro spherical virus reveals evidence of genetic recombination between distinct isolates.  

PubMed

In this study, complete genome of a south Indian isolate of Rice tungro spherical virus (RTSV) from Andhra Pradesh (AP) was sequenced, and the predicted amino acid sequence was analysed. The RTSV RNA genome consists of 12,171 nt without the poly(A) tail, encoding a putative typical polyprotein of 3,470 amino acids. Furthermore, cleavage sites and sequence motifs of the polyprotein were predicted. Multiple alignment with other RTSV isolates showed a nucleotide sequence identity of 95% to east Indian isolates and 90% to Philippines isolates. A phylogenetic tree based on complete genome sequence showed that Indian isolates clustered together, while Vt6 and PhilA isolates of Philippines formed two separate clusters. Twelve recombination events were detected in RNA genome of RTSV using the Recombination Detection Program version 3. Recombination analysis suggested significant role of 5' end and central region of genome in virus evolution. Further, AP and Odisha isolates appeared as important RTSV isolates involved in diversification of this virus in India through recombination phenomenon. The new addition of complete genome of first south Indian isolate provided an opportunity to establish the molecular evolution of RTSV through recombination analysis and phylogenetic relationship. PMID:23925555

Sailaja, B; Anjum, Najreen; Patil, Yogesh K; Agarwal, Surekha; Malathi, P; Krishnaveni, D; Balachandran, S M; Viraktamath, B C; Mangrauthia, Satendra K

2013-12-01

271

Influenza A viruses isolated from waterfowl in two wildlife management areas of Pennsylvania.  

PubMed

A survey was conducted at two wildlife management areas of Pennsylvania (USA) to evaluate an antigen capture enzyme-linked immunosorbent assay (AC-ELISA) for the detection of avian influenza viruses (AIV) in cloacal swabs from waterfowl and to determine the influenza A virus subtypes and the distribution of these viruses among waterfowl. We collected 330 cloacal swabs from hunter-killed waterfowl in the fall of 1990 and from cage-captured waterfowl in the summer of 1991. Thirty-one hemagglutinating agents were isolated by chicken embryo inoculation (CEI) of which 27 were influenza A viruses and four Newcastle disease viruses (NDV). The prevalence of AIV infection was 8.2%. Compared to CEI, AC-ELISA was only 15% sensitive and 61% specific. Based on the distribution of AIV by species of waterfowl, mallards (Anas platyrhynchos) and American wigeons (Anas americana) were at equal risk of AIV infection even though most of the AIV isolates came from mallards. Although significant crude effects of sampling site and season on AIV recovery could be established, juvenile age was identified as the primary risk factor of AIV recovery. Twelve AIV subtypes were identified by hemagglutination inhibition (HI) and neuraminidase inhibition (NI) tests. The most prevalent subytpes were H4N8 and H6N8. We concluded that AC-ELISA was not useful for the detection of AIV in cloacal swabs from waterfowl and that CEI, HI, and NI tests remain as the method of choice for AIV screening in waterfowl. Based on the results AIV infected preferentially the young which represent the high risk group in waterfowl populations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8583635

Alfonso, C P; Cowen, B S; van Campen, H

1995-04-01

272

Isolation of midgut escape mutants of two American genotype dengue 2 viruses from Aedes aegypti  

PubMed Central

Background Several studies have shown that American genotype dengue 2 viruses (DENV2) have reduced viral fitness in the mosquito vector, Aedes aegypti, compared to other DENV2 genotypes. Diminished replication efficiency or inability to efficiently traverse membrane barriers encompassing organs such as the midgut or salivary glands are considered major factors negatively impacting viral fitness in the mosquito. Results We analyzed the vector competence of Ae. aegypti for two American DENV2 strains, QR94 and PR159 originating from Mexico and Puerto-Rico, respectively. Both strains infected mosquito midguts following acquisition of infectious bloodmeals. However, DENV2-QR94 and DENV2-PR159 poorly disseminated from the midgut at 7 or 14 days post-bloodmeal (pbm). We detected one virus isolate, EM33, among 31 DENV2-QR94 infected mosquitoes, and one isolate, EM41, among 121 DENV2-PR159 infected mosquitoes, generating high virus titers in mosquito carcasses at 7 days pbm. In oral challenge experiments, EM33 and EM41 showed midgut dissemination rates of 40-50%. Replication efficiency of EM41 in secondary mosquito tissue was similar to that of a dissemination-competent control strain, whereas the replication efficiency of EM33 was significantly lower than that of the control virus. The genome sequence of DENV2-QR94 encoded seven unique amino acids (aa), which were not found in 100 of the most closely related DENV2 strains. EM33 had one additional aa change, E202K, in the E protein. DENV2-PR159 encoded four unique aa residues, one of them E202K, whereas EM41 had two additional aa substitutions, Q77E in the E protein and E93D in NS3. Conclusions Our results indicate that the midgut of Ae. aegypti acts as a selective sieve for DENV2 in which genetically distinct, dissemination-competent virus variants are rapidly selected from the viral quasispecies to be transmitted to vertebrates. PMID:23937713

2013-01-01

273

Complete genome sequence of a novel porcine parainfluenza virus 5 isolate in Korea.  

PubMed

A novel cytopathogenic paramyxovirus was isolated from a lung sample from a piglet, using continuous porcine alveolar macrophage cells. Morphologic and genetic studies indicated that this porcine virus (pPIV5) belongs to the species Parainfluenza 5 in the family Paramyxoviridae. We attempted to determine the complete nucleotide sequence of the first Korean pPIV5 isolate, designated KNU-11. The full-length genome of KNU-11 was found to be 15,246 nucleotides in length and consist of seven nonoverlapping genes (3'-N-V/P-M-F-SH-HN-L-5') predicted to encode eight proteins. The overall degree of nucleotide sequence identity was 98.7 % between KNU-11 and PIV5 (formerly simian virus 5, SV5), a prototype paramyxovirus, and the putative proteins had 74.4 to 99.2 % amino acid identity to those of PIV5. Phylogenetic analysis further demonstrated that the novel pPIV5 isolate is a member of the genus Rubulavirus of the subfamily Paramyxovirinae. The present study describes the identification and genomic characterization of a pPIV5 isolate in South Korea. PMID:23807746

Lee, Yu Na; Lee, Changhee

2013-08-01

274

Molecular identification of Cucumber mosaic virus isolates of subgroup IB associated with mosaic disease of eggplant in India.  

PubMed

Association of Cucumber mosaic virus (CMV) with severe mosaic disease of eggplant (Solanum melongena L.) collected from Lucknow and Kanpur, India was initially detected by host reaction and serological assay and confirmed by RT-PCR employing coat protein gene specific primers. Further, molecular identification of the virus isolates was done by cloning and sequence analysis of the complete RNA3 genome. Based on 97-99 % identities and phylogenetic relationships, the virus isolates infecting eggplant were identified as members of CMV subgroup IB. PMID:24426321

Kumar, Susheel; Gautam, Karmveer Kumar; Raj, Shri Krishna

2014-01-01

275

Differentiation of field isolates and vaccine strains of infectious laryngotracheitis virus by DNA sequencing.  

PubMed

Two different regions of the infected cell protein 4 (ICP4) gene of infectious laryngotracheitis virus (ILTV) were amplified and sequenced for characterization of field isolates and tissue culture-origin (TCO) and chicken embryo-origin (CEO) vaccine strains. Phylogenetic analysis of the two regions showed differences in nucleotide and amino acid sequences between field isolates and attenuated vaccines. The PCR-RFLP results were identical to those obtained by DNA sequencing and validated their use to differentiate ILTV strains. The approach using the sequencing of the two fragments of the ICP4 gene showed to be an efficient and practical procedure to differentiate between field isolates and vaccine strains of ILTV. PMID:19747995

Chacón, Jorge Luis; Ferreira, Antonio J Piantino

2009-11-12

276

Demonstration of antigenic variation among rabies virus isolates by using monoclonal antibodies to nucleocapsid proteins.  

PubMed Central

Rabies virus isolates from terrestrial animals in six areas of the United States were examined with a panel of monoclonal antibodies to nucleocapsid proteins. Characteristic differences in immunofluorescence reactions permitted the formation of four antigenically distinct reaction groups from the 231 isolates tested. The geographic distribution of these groups corresponded well with separate rabies enzootic areas recognized by surveillance of sylvatic rabies in the United States. Distinctive reaction patterns were also identified for viral proteins from four infected bat species, and identical patterns were found in eight isolated cases of rabies in terrestrial animals. These findings suggest that monoclonal antibodies can be used to study the prevalence, distribution, and transmission of rabies among wildlife species. PMID:2429983

Smith, J S; Reid-Sanden, F L; Roumillat, L F; Trimarchi, C; Clark, K; Baer, G M; Winkler, W G

1986-01-01

277

Restriction endonuclease analysis of bovine herpesvirus type 1 isolates from calves with fatal encephalitis: comparison with vaccine virus.  

PubMed

Meningo-encephalitis in feedlot cattle sporadically occurred in the Tokachi area in northern Japan. The calves had been vaccinated intranasally with a mixed live-vaccine (infectious bovine rhinotracheitis virus, bovine viral diarrhea-mucosal disease virus, and parainfluenza 3 virus) for which intramuscular inoculation was indicated. Two additional live vaccines, bovine adenovirus type 7 and bovine respiratory syncytical virus, had been inoculated simultaneously. Eleven isolates of bovine herpesvirus type 1 were plaque-purified from two brains with fatal encephalitis; their viral DNAs were examined by restriction endonuclease analysis (REA) using PstI and HindIII. The REA patterns of the virus clones were almost identical to those of the vaccine strains 758-43, suggesting that the isolates from this outbreak of fatal encephalitis originated in the abnormally administered vaccine. PMID:7548427

Horiuchi, M; Yamazaki, N; Furuoka, H; Matsui, T; Nakagawa, M; Ishiguro, N; Shinagawa, M

1995-06-01

278

Single Strain Isolation Method for Cell Culture-Adapted Hepatitis C Virus by End-Point Dilution and Infection  

PubMed Central

The hepatitis C virus (HCV) culture system has enabled us to clarify the HCV life cycle and essential host factors for propagation. However, the virus production level of wild-type JFH-1 (JFH-1/wt) is limited, and this leads to difficulties in performing experiments that require higher viral concentrations. As the cell culture-adapted JFH-1 has been reported to have robust virus production, some mutations in the viral genome may play a role in the efficiency of virus production. In this study, we obtained cell culture-adapted virus by passage of full-length JFH-1 RNA-transfected Huh-7.5.1 cells. The obtained virus produced 3 log-fold more progeny viruses as compared with JFH-1/wt. Several mutations were identified as being responsible for robust virus production, but, on reverse-genetics analysis, the production levels of JFH-1 with these mutations did not reach the level of cell culture-adapted virus. By using the single strain isolation method by end-point dilution and infection, we isolated two strains with additional mutations, and found that these strains have the ability to produce more progeny viruses. On reverse-genetics analysis, the strains with these additional mutations were able to produce robust progeny viruses at comparable levels as cell culture-adapted JFH-1 virus. The strategy used in this study will be useful for identifying strains with unique characteristics, such as robust virus production, from a diverse population, and for determining the responsible mutations for these characteristics. PMID:24848954

Sugiyama, Nao; Murayama, Asako; Suzuki, Ryosuke; Watanabe, Noriyuki; Shiina, Masaaki; Liang, T. Jake; Wakita, Takaji; Kato, Takanobu

2014-01-01

279

Detection and isolation of infectious laryngotracheitis virus on a broiler farm after a disease outbreak.  

PubMed

A broiler farm in North Alabama suffered a mild infectious laryngotracheitis (ILT) outbreak, as determined by clinical disease and PCR. The poultry integrator sought help to control further outbreaks in subsequent flocks. Samples were collected from various areas of the poultry houses on the farm over an 8-wk period. The first sampling was conducted 8 days after the infected farm was depopulated; the second was conducted 2 days prior to subsequent flock placement; and the third was conducted when the new flock was 5 wk of age. Samples were examined for ILT virus (ILTV) DNA by real-time PCR and virus isolation in embryos. The infected houses were cleaned, disinfected, heated, litter composted, and curtains replaced after the first sampling and prior to placement of the next flock. Samples from all periods were positive for ILTV DNA. However, the number of positive samples and crossing point values indicated a decrease in the amount of viral DNA, while virus isolation in embryos was successful only on the first sampling. The subsequent flock was vaccinated against ILTV by in ovo route using a commercial recombinant vaccine. Cleaning and sanitation after the disease outbreak reduced the amount of ILTV on the farm and together with in ovo vaccination of the new flock may have prevented a recurrence of another ILT outbreak. PMID:24597126

Dormitorio, Teresa V; Giambrone, Joseph J; Macklin, Kenneth S

2013-12-01

280

Genomic characterization of a bovine viral diarrhea virus 1 isolate from swine.  

PubMed

The SD0803 strain of the bovine viral diarrhea virus (BVDV) was isolated from a piglet in China in 2008 and has been classified as a novel subgenotype of BVDV-1. To describe the molecular features of this novel subgenotype, we sequenced and characterized the complete genome of the SD0803 virus. The genome is 12,271 bp in length and contains 5' and 3' untranslated regions (UTRs) that flank an open reading frame (ORF) encoding a 3,898-amino-acid polypeptide. The full-length genome of the SD0803 strain shares 78.8% to 83.3% identity with those of other BVDV-1 strains, 70.0% to 70.7% identity with those of BVDV-2 strains, and less than 67.6% identity with those of other pestiviruses. The highest level of shared identity was 83.3% between the complete SD0803 genome and that of the ZM-95 strain of BVDV-1. Phylogenetic analysis of the 5' UTR and the coding sequence for the N-terminal protease fragment of the SD0803 polyprotein indicated that the SD0803 virus is a member of the novel subgenotype BVDV-1q, isolates of which have been identified recently in dairy cattle and camels in China. PMID:24719194

Deng, Yu; Shan, Tong-Ling; Tong, Wu; Zheng, Xu-Chen; Guo, Yi-Yei; Zheng, Hao; Cao, San-Jie; Wen, Xin-Tian; Tong, Guang-Zhi

2014-09-01

281

Isolation of Maedi/Visna Virus from a Sheep in Japan  

PubMed Central

ABSTRACT Maedi/visna (MV) is a lentiviral disease of sheep caused by the maedi/visna virus (MVV). Although MV is prevalent in many countries, it had not been reported in Japan. In 2011, however, three sheep in northern Japan were reported to be seropositive against the MVV antigen, indicating a persistent MVV infection. In the present study, we isolated MVV from one sheep to confirm MVV infection and conducted genomic classification of the virus. The co-culture of leukocytes from a seropositive sheep with fetal goat lung cells resulted in the formation of syncytial cells and the amplification of a long terminal repeat sequence of MVV by polymerase chain reaction. The isolate was confirmed as being MVV, rather than the caprine arthritis-encephalitis virus based on phylogenetic analysis of the gag gene sequence. Although the sheep was asymptomatic, nonpurulent meningitis and demyelination were found in the spinal cord. These were considered to be early lesions associated with pathogenic MVV infection. Therefore, the present study demonstrated that MVV is distributed in Japan. PMID:24141278

OGUMA, Keisuke; TANAKA, Chiaki; HARASAWA, Ryo; KIMURA, Atsushi; SASAKI, Jun; GORYO, Masanobu; SENTSUI, Hiroshi

2013-01-01

282

Isolation of maedi/visna virus from a sheep in Japan.  

PubMed

Maedi/visna (MV) is a lentiviral disease of sheep caused by the maedi/visna virus (MVV). Although MV is prevalent in many countries, it had not been reported in Japan. In 2011, however, three sheep in northern Japan were reported to be seropositive against the MVV antigen, indicating a persistent MVV infection. In the present study, we isolated MVV from one sheep to confirm MVV infection and conducted genomic classification of the virus. The co-culture of leukocytes from a seropositive sheep with fetal goat lung cells resulted in the formation of syncytial cells and the amplification of a long terminal repeat sequence of MVV by polymerase chain reaction. The isolate was confirmed as being MVV, rather than the caprine arthritis-encephalitis virus based on phylogenetic analysis of the gag gene sequence. Although the sheep was asymptomatic, nonpurulent meningitis and demyelination were found in the spinal cord. These were considered to be early lesions associated with pathogenic MVV infection. Therefore, the present study demonstrated that MVV is distributed in Japan. PMID:24141278

Oguma, Keisuke; Tanaka, Chiaki; Harasawa, Ryo; Kimura, Atsushi; Sasaki, Jun; Goryo, Masanobu; Sentsui, Hiroshi

2014-03-01

283

Isolation and replication of rabies virus in C6 rat glioma cells (clone CCL-107).  

PubMed

The susceptibility of the C6 rat glioma cell line (ATCC; CCL-107) to rabies virus was characterized. The kinetics of infection performed with a fixed and a wild strain (from an infected cow) of rabies virus was monitored by direct immunofluorescence. Fluorescent cytoplasmic bodies were readily observed by UV microscopy from 24 hours post-infection (hpi) onwards. The ability of C6 to produce rabies infective virion particles was confirmed by determining the viral titres present in the supernatants of infected cultures, by both BHK-21 cell infection and mice inoculation. C6 cells produced similar viral titres to those produced by BHK-21 for both strains used. In addition, the yield of rabies glycoprotein was assessed by ELISA. In general, BHK-21 and C6 cells infected either by PV or with the wild rabies strain produced similar amounts of rabies glycoprotein. At 96 hpi, however, when the glycoprotein production peaked, BHK-21 infected with the wild strain produced significantly higher amounts of glycoprotein than C6. Subsequently, the optimal conditions for isolation of wild rabies virus strains from C6 cells were established and these proved to be as sensitive as NA cells in detecting 10 wild rabies samples. Due to the high sensitivity exhibited, C6 rat glioma cells present a new and useful system for rabies virus investigation. PMID:11580211

Bordignon, J; Piza, A T; Alvarez-Silva, M; Caporale, G M; Carrieri, M L; Kotait, I; Zanetti, C R

2001-06-01

284

Phosphonoacetic Acid-Resistant Mutants of Herpes Simplex Virus: Effect of Phosphonoacetic Acid on Virus Replication and In Vitro Deoxyribonucleic Acid Synthesis in Isolated Nuclei  

PubMed Central

Phosphonoacetic acid (PAA) inhibits the replication of herpes simplex virus in BSC-1 cells and the in vitro synthesis of deoxyribonucleic acid (DNA) in isolated nuclei. Phosphonopropionic acid at a concentration of 100 ?g/ml had no effect on herpes simplex virus replication. PAA-resistant mutants were obtained at a rate of 1 in 104 plaque-forming units after 5-bromodeoxyuridine mutagenization of the virus. These mutants replicate in BSC-1 cells in the presence of 100 ?g of PAA per ml and induce a PAA-resistant DNA polymerase that synthesizes DNA in vitro in the presence of PAA. PMID:195514

Becker, Yechiel; Asher, Yael; Cohen, Yaffa; Weinberg-Zahlering, Eynat; Shlomai, Joseph

1977-01-01

285

Biochemical and Antigenic Properties of the First Isolates of Infectious Hematopoietic Necrosis Virus from Salmonid Fish in Europe  

Microsoft Academic Search

The first isolates of infectious hematopoietic necrosis virus (IHNV) recovered from rainbow trout Oncorhynchus mykiss (formerly Salmo gairdneri) in France and Italy were compared to six representative strains from North America by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of virion polypeptides and neutralization by monoclonal antibodies (MAbs). All three IHNV isolates from Europe had similar polypeptide profiles when compared by

K. D. Arkush; G. Bovo; P. De Kinkelin; J. R. Winton; W. H. Wingfield; R. P. Hedrick

1989-01-01

286

Whole-Genome Sequence of a Reassortant H5N6 Avian Influenza Virus Isolated from a Live Poultry Market in China, 2013  

PubMed Central

An avian influenza virus, A/environment/Zhenjiang/C13/2013(H5N6), was isolated from a live poultry market in eastern China. Phylogenetic analysis showed that the isolate was a novel reassortant virus with a neuraminidase (NA) gene from H6N6 viruses and the other seven genes from H5N1 viruses, which may pose a potential threat to human and animal health. PMID:25212611

Qi, Xian; Cui, Lunbiao; Yu, Huiyan; Ge, Yiyue

2014-01-01

287

Identification of Recombination in the Envelope Gene of Simian Foamy Virus Serotype 2 Isolated from Macaca cyclopis  

PubMed Central

The full-length sequence of simian foamy virus serotype 2 (SFVmcy-2), isolated from a Taiwanese macaque, was determined. SFVmcy-2 was highly related to SFV serotype 1 (SFVmcy-1), an isolate from the same species, except in the putative receptor binding domain (RBD) in env, which contained novel sequences related to SFV serotype 3 (SFVagm-3), isolated from an African green monkey. The results identify a potential region of neutralization in SFVs and demonstrate recombination between genetically divergent foamy viruses. PMID:23698303

Galvin, Teresa A.; Ahmed, Imran A.; Shahabuddin, Muhammad; Bryan, Theodore

2013-01-01

288

Infectivity and complete nucleotide sequence of cucumber fruit mottle mosaic virus isolate Cm cDNA.  

PubMed

Three isolates of cucumber fruit mottle mosaic virus (CFMMV) were collected from melon, cucumber, and pumpkin plants in Korea. A full-length cDNA clone of CFMMV-Cm (melon isolate) was produced and evaluated for infectivity after T7 transcription in vitro (pT7CF-Cmflc). The complete CFMMV genome sequence of the infectious clone pT7CF-Cmflc was determined. The genome of CFMMV-Cm consisted of 6,571 nucleotides and shared high nucleotide sequence identity (98.8 %) with the Israel isolate of CFMMV. Based on the infectious clone pT7CF-Cmflc, a CaMV 35S-promoter driven cDNA clone (p35SCF-Cmflc) was subsequently constructed and sequenced. Mechanical inoculation with RNA transcripts of pT7CF-Cmflc and agro-inoculation with p35SCF-Cmflc resulted in systemic infection of cucumber and melon, producing symptoms similar to those produced by CFMMV-Cm. Progeny virus in infected plants was detected by RT-PCR, western blot assay, and transmission electron microscopy. PMID:24473709

Rhee, Sun-Ju; Hong, Jin-Sung; Lee, Gung Pyo

2014-07-01

289

Isolation, molecular characterization, and phylogenetic analysis of encephalomyocarditis virus from South China tigers in China.  

PubMed

Although encephalomyocarditis virus (EMCV) can infect many host species and cause myocarditis and sudden death in many species, little is known about EMCV infection in tigers. A virus was isolated from organs of dead South China tigers with sudden death in southern China. The production of cytopathic effect on BHK cells, and the results of PCR, electron microscopy (EM), and whole genome sequencing indicated that the pathogen was EMCV, the strain was named FJ13. Other pathogenic agents were excluded as possible pathogenic agents. Phylogenetic analyses of the whole genome, ORF (open reading frame) and CCR (capsid coding region) using the neighbour-joining method revealed that EMCV isolates cluster into two groups (group 1 and 2) with two sub-clusters within group 1 (group 1a and 1b), and FJ13 belongs to group 1a. Animal experiment showed that the isolated strain FJ13 could cause clinical symptoms and pathological changes. The results of this study indicated that FJ13 caused myocarditis of tigers and provided new epidemiologic data on EMCV in China. PMID:23917023

Liu, Huimin; Yan, Qi; Zhao, Bo; Luo, Jing; Wang, Chengmin; Du, Yingchun; Yan, Jing; He, Hongxuan

2013-10-01

290

Characteristics of a Lettuce mosaic virus Isolate Infecting Lettuce in Korea  

PubMed Central

Lettuce mosaic virus (LMV) causes disease of plants in the family Asteraceae, especially lettuce crops. LMV isolates have previously been clustered in three main groups, LMV-Yar, LMV-Greek and LMVRoW. The first two groups, LMV-Yar and LMV-Greek, have similar characteristics such as no seed-borne transmission and non-resistance-breaking. The latter one, LMV-RoW, comprising a large percentage of the LMV isolates contains two large subgroups, LMV-Common and LMV-Most. To date, however, no Korean LMV isolate has been classified and characterized. In this study, LMV-Muju, the Korean LMV isolate, was isolated from lettuce showing pale green and mottle symptoms, and its complete genome sequence was determined. Classification method of LMV isolates based on nucleotide sequence divergence of the NIb-CP junction showed that LMV-Muju was categorized as LMV-Common. LMV-Muju was more similar to LMV-O (LMV-Common subgroup) than to LMV-E (LMV-RoW group but not LMV-Common subgroup) even in the amino acid domains of HC-Pro associated with pathogenicity, and in the CI and VPg regions related to ability to overcome resistance. Taken together, LMV-Muju belongs to the LMV-Common subgroup, and is expected to be a seed-borne, non-resistance-breaking isolate. According to our analysis, all other LMV isolates not previously assigned to a subgroup were also included in the LMV-RoW group.

Lim, Seungmo; Zhao, Fumei; Yoo, Ran Hee; Igori, Davaajargal; Lee, Su-Heon; Lim, Hyoun-Sub; Moon, Jae Sun

2014-01-01

291

Complete genomic sequence of a Tobacco rattle virus isolate from Michigan-grown potatoes.  

PubMed

Tobacco rattle virus (TRV) causes stem mottle on potato leaves and necrotic arcs and rings in potato tubers, known as corky ringspot disease. Recently, TRV was reported in Michigan potato tubers cv. FL1879 exhibiting corky ringspot disease. Sequence analysis of the RNA-1-encoded 16-kDa gene of the Michigan isolate, designated MI-1, revealed homology to TRV isolates from Florida and Washington. Here, we report the complete genomic sequence of RNA-1 (6,791 nt) and RNA-2 (3,685 nt) of TRV MI-1. RNA-1 is predicted to contain four open reading frames, and the genome structure and phylogenetic analyses of the RNA-1 nucleotide sequence revealed significant homologies to the known sequences of other TRV-1 isolates. The relationships based on the full-length nucleotide sequence were different from than those based on the 16-kDa gene encoded on genomic RNA-1 and reflect sequence variation within a 20-25-aa residue region of the 16-kDa protein. MI-1 RNA-2 is predicted to contain three ORFs, encoding the coat protein (CP), a 37.6-kDa protein (ORF 2b), and a 33.6-kDa protein (ORF 2c). In addition, it contains a region of similarity to the 3' terminus of RNA-1, including a truncated portion of the 16-kDa cistron. Phylogenetic analysis of RNA-2, based on a comparison of nucleotide sequences with other members of the genus Tobravirus, indicates that TRV MI-1 and other North American isolates cluster as a distinct group. TRV M1-1 is only the second North American isolate for which there is a complete sequence of the genome, and it is distinct from the North American isolate TRV ORY. The relationship of the TRV MI-1 isolate to other tobravirus isolates is discussed. PMID:20221838

Crosslin, James M; Hamm, Philip B; Kirk, William W; Hammond, Rosemarie W

2010-04-01

292

Phylogenetic analyses of highly pathogenic avian influenza virus isolates from Germany in 2006 and 2007 suggest at least three separate introductions of H5N1 virus.  

PubMed

In spring 2006, highly pathogenic avian influenza virus (HPAIV) of subtype H5N1 was detected in Germany in 343 dead wild birds, as well as in a black swan (Cygnus atratus) kept in a zoo, three stray cats, one stone marten (Martes foina), and in a single turkey farm. In 2007 (June-July) the virus reoccurred in 96 wild birds at six geographically separate locations in the Southeast of Germany. In addition, a backyard mixed duck and goose holding was affected. Real-time RT-PCR [Hoffmann, B., Harder, T., Starick, E., Depner, K., Werner, O., Beer, M., 2007. Rapid and highly sensitive pathotyping of avian influenza A H5N1 virus by using real-time reverse transcription-PCR. J. Clin. Microbiol. 45, 600-603] and nucleotide sequencing confirmed that these H5-viruses belonged to the Qinghai lineage of HPAIV H5N1 (clade 2.2). For a more detailed analysis, the hemagglutinin and neuraminidase genes of 27 selected German H5N1 viruses isolated 2006 or 2007 and originating from different regions and animal species were sequenced and analysed phylogenetically. As a result, three closely related but distinguishable H5N1 subclades could be defined: In 2006 a 'Northern type' (subclade 2.2.2), representing virus isolates from the German federal states Mecklenburg-Western Pomerania, Schleswig-Holstein, Brandenburg, and Lower Saxony, and a 'Southern type' (subclade 2.2.1) from Baden-Württemberg and Bavaria were detected. Interestingly, representatives of both types were present in Central Germany and caused the outbreak in turkeys (subclade 2.2.2) and in a case in a tufted duck (Aythya fuligula) (subclade 2.2.1) in Saxony. Furthermore, one isolate from the South of Germany was identified as 2.2.2 and vice versa a 2.2.1-like isolate was found in Northern Germany. H5N1 viruses isolated in 2007 belonged to a third type (subclade 2.2.3) which was not detected in 2006. Our data suggest the introduction of three distinct H5N1 variants into the wild bird population of Germany. The source of these viruses and the exact time of introduction remain obscure. Based on the identification of closely related H5N1 viruses from Southern and Central Russia, a recent introduction via wild birds on winter escape from these regions, early in 2006 constitutes the most likely scenario for the 2006 outbreaks. The viruses detected in 2007 most likely represent another new incursion from an as yet unknown source. PMID:18031958

Starick, E; Beer, M; Hoffmann, B; Staubach, C; Werner, O; Globig, A; Strebelow, G; Grund, C; Durban, M; Conraths, F J; Mettenleiter, T; Harder, T

2008-04-30

293

The genomic RNA1 and RNA2 sequences of the tobacco rattle virus isolates found in Polish potato fields.  

PubMed

Four tobacco rattle virus (TRV) isolates were identified from tobacco bait seedlings planted in soil samples from Polish potato fields. Sequence analysis of the genomic RNA1 of the isolates revealed significant similarity to the isolates Ho and AL recently found in Germany. Multiple sequence alignments of the genomic RNA2 indicated that the two isolates from northern Poland (Deb57 and Slu24) are in a cluster with the isolates PSG and PLB found in the Netherlands. The remaining two isolates, from central Poland (11r21 and Mlo7), are in a distinct group with the unique isolate SYM found in England. The RNA2 sequences of the studied isolates range from 1998 nt to 2739 nt in length, and all carry deletions of the 2b and/or 2c genes. The isolate Mlo7 has an atypical RNA2 structure, having its cp gene located in its central region. PMID:24637409

Yin, Zhimin; Pawe?kowicz, Magdalena; Michalak, Krystyna; Chrzanowska, Miros?awa; Zimnoch-Guzowska, Ewa

2014-06-24

294

Isolation and propagation of Dengue virus in Vero and BHK-21 cells expressing human DC-SIGN stably.  

PubMed

The "standard" methods of isolating dengue virus (DENV) utilize the mosquito cell line C6/36, monkey kidney LLC-MK2 cells, Vero cells, or baby hamster kidney (BHK-21) cells. However, these cells lines lack a particular DENV receptor, known as dendritic cell-specific ICAM-3-grabbing non-integrin (DC-SIGN), which is expressed on immature dendritic cells and monocytes/macrophages. This may result in less efficient virus isolation and propagation. The present study used a lentivirus vector to establish Vero and BHK-21 cell lines (Vero-DC and BHK-DC) that express human DC-SIGN stably. Five DENV strains, each passaged several times in C6/36 cells, replicated more efficiently in Vero-DC and BHK-DC than in the parental Vero or BHK-21 cells. Vero/Vero-DC and BHK-21/BHK-DC were used to isolate virus from buffy coats and plasma samples derived from 13 patients infected with DENV. Most of the viruses showed increased production in cell lines expressing DC-SIGN. However, the isolation rate was lower (15.4-46.2%) than that from C6/36 cells (84.6%). Interestingly, when the viruses were isolated in C6/36 cells prior to infecting Vero/Vero-DC and BHK-21/BHK-DC, the rate of virus production increased markedly, reaching levels higher than those initially achieved in C6/36 cells. These data suggest that Vero-DC and BHK-DC could be useful tools for virus propagation, and that human specimens may contain a factor that interferes with virus growth in mammalian cells. PMID:25205264

Phanthanawiboon, Supranee; A-Nuegoonpipat, Atchareeya; Panngarm, Narawan; Limkittikul, Kriengsak; Ikuta, Kazuyoshi; Anantapreecha, Surapee; Kurosu, Takeshi

2014-12-01

295

First isolation and molecular characterization of foot-and-mouth disease virus in Benin.  

PubMed

Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals. It is one of the most economically devastating diseases affecting livestock animals. In West Africa, where constant circulation of FMD virus (FMDV) is assumed, very few studies on the characterization of circulating strains have been published. This study describes the first isolation and characterization of FMDV in Benin. FMDV was isolated from 42 samples. Antigen Capture Elisa (Ag-ELISA) and VP1 coding sequence analysis revealed 33 strains of serotype O and 9 strains of serotype A. Phylogenetic analysis of the VP1 sequence revealed two different groups of type O isolates and one group of A isolates. VP1 sequence comparison with the sequences available in the GenBank database revealed a close relationship of the Benin isolates with topotype O of West Africa and with African topotype A of genotype VI. Knowledge of the recent strains circulating in Benin should contribute to better selection of vaccine strains and enable the updating of molecular epidemiology data available for West Africa in general. PMID:24720890

Gorna, Kamila; Houndjč, Evelyne; Romey, Aurore; Relmy, Anthony; Blaise-Boisseau, Sandra; Kpodékon, Marc; Saegerman, Claude; Moutou, François; Zientara, Stephan; Bakkali Kassimi, Labib

2014-06-25

296

Isolation and characterization of the dsRNA virus from the yeast Endomyces magnusii.  

PubMed

Virus-like particles (VLPs) have been isolated from the yeast Endomyces magnusii. The VLPs measure 43 nm in diameter and contain six species of dsRNA (0.78, 0.83, 1.77, 1.84, 2.64, 4.30 kb respectively). E. magnusii produces a 'toxic' protein, which reduces the growth, and changes the colony morphology, of sensitive strains of Hansenula sp. growing on solid media. All strains of E. magnusii tested produced the 'toxin' and contained the VLPs. Current procedures of curing failed to destroy the ability to produce the 'toxin'. PMID:8150269

Pospísek, M; Palková, Z; Janderová, B; Korb, J

1994-02-15

297

Isolation and characterization of a new strain of lymphocytic choriomeningitis virus from rodents in southwestern France.  

PubMed

A total of 821 tissue samples from rodents trapped during field campaigns organized in Europe and Africa were screened for the presence of arenaviruses by molecular methods and cell culture inoculation when feasible. Two Mus musculus domesticus trapped in the southwestern part of France were infected with a potentially new strain of lymphocytic choriomeningitis virus (LCMV), here referred to as LCMV strain HP65-2009, which was isolated and genetically characterized by whole genome sequencing. Genetic and phylogenetic analyses comparing LCMV HP65-2009 with 26 other LCMV strains showed that it represents a novel highly-divergent strain within the group of Mus musculus-associated LCMV. PMID:22651393

Yama, Ines N; Cazaux, Benoite; Britton-Davidian, Janice; Moureau, Grégory; Thirion, Laurence; de Lamballerie, Xavier; Dobigny, Gauthier; Charrel, Rémi N

2012-10-01

298

Complete genome analysis of a novel recombinant isolate of potato virus Y from China.  

PubMed

The complete sequence of GF_YL20, a potato virus Y (PVY) isolate from China, encodes a polyprotein of 3,061 amino acids. Sequence analysis indicates that GF_YL20 has a genomic structure different from previously reported PVY strains. It shares 99 % nucleotide sequence identity with PB209 (PVY(N:O)) except in VPg, but more than 97 % nucleotide sequence identity with the VPg of Mont (PVY(N)), PB312 (PVY(NTN)) and HN2 (SYR-I). Phylogenetic analysis indicates that GF_YL20 is a novel N:O recombinant with three recombination breakpoints. PMID:25091741

Gao, Fangluan; Chang, Fei; Shen, Jianguo; Shi, Fengyang; Xie, Lianhui; Zhan, Jiasui

2014-12-01

299

[The isolation of Dhori viruses (Orthomyxoviridae, Thogotovirus) and Crimean-Congo hemorrhagic fever virus (Bunyaviridae, Nairovirus) from the hare (Lepus europaeus) and its ticks Hyalomma marginatum in the middle zone of the Volga delta, Astrakhan region, 2001].  

PubMed

In August, 2001, in the middle zone of the delta of the Volga River, the Astrakhan region, during investigation of the natural foci of West Nile fever and Crimean--Congo hemorrhagic fever (CCHF), the material from the hare (Lepus europaeus, Pallas, 1778 (Lagomorpha, Leporidae) and collected from it the ticks Hyalomna marginatum Koch 1844, was obtained. 4 strains of Dhori virus (Orthomyxoviridae, Thogotovirus) and 2 strains of CCHF virus (Bunyaviridae, Nairovirus) were isolated. This is the first isolation of Thogotovirus genus virus from the wild vertebrates. Considering the overlap of the Dhori virus and CCHF virus areas, similar ecology and the isolation both viruses from the same pool of the ticks, the necessity for the use of the test-system for indication of the viruses, differential diagnosis and accumulation of the data concerning the role of Dhori virus in the human and farm animals pathology is discussed. PMID:12271723

L'vov, D N; Dzharkenov, A F; Aristova, V A; Kovtunov, A I; Gromashevski?, V L; Vyshemirski?, O I; Galkina, I V; Larichev, V F; Butenko, A M; L'vov, D K

2002-01-01

300

Study of oseltamivir and zanamivir resistance-related mutations in influenza viruses isolated from wild mallards in Sweden.  

PubMed

Resistance to neuraminidase inhibitors (NAIs) is a growing problem in battle against influenza A virus. However, little is known about the resistance of viruses isolated from dabbling ducks, the natural reservoir of the influenza virus. To our knowledge, no low-pathogenic avian influenza (LPAI) virus resistant to NAIs has been detected. The aim of this study was to investigate mallard isolates of influenza A virus previously identified to carry oseltamivir carboxylate (OC) or zanamivir (ZA) resistance-related mutations. In this work, 21 viruses belonging to the N1, N3, N6 and N9 subtypes were analyzed using a colorimetric NA inhibition assay. The results of assay showed no NAIs-resistant phenotype for any of the viruses. The R118K mutation was the most recurrent, as it was observed in all subtypes except for N6. IC50 values confirmed the differences in sensitivity to OC or ZA observed in the N1 and N2 groups of NAs. Furthermore, both wild types (WTs) in the N6 and one WT in the N9 subtype were less sensitive to ZA than were genotypically related mutants with R152K and R118K change in the respective subtypes. This may indicate that these and probably even other NAIs resistance-related mutations found in our virus collection were not induced by NAIs residuals in the environment and that the impact of such mutations in an avian influenza could be dependent on subtype, strain and host species. PMID:24558492

Orozovic, Goran; Orozovic, Kanita; Järhult, Josef D; Olsen, Björn

2014-01-01

301

Study of Oseltamivir and Zanamivir Resistance-Related Mutations in Influenza Viruses Isolated from Wild Mallards in Sweden  

PubMed Central

Resistance to neuraminidase inhibitors (NAIs) is a growing problem in battle against influenza A virus. However, little is known about the resistance of viruses isolated from dabbling ducks, the natural reservoir of the influenza virus. To our knowledge, no low-pathogenic avian influenza (LPAI) virus resistant to NAIs has been detected. The aim of this study was to investigate mallard isolates of influenza A virus previously identified to carry oseltamivir carboxylate (OC) or zanamivir (ZA) resistance-related mutations. In this work, 21 viruses belonging to the N1, N3, N6 and N9 subtypes were analyzed using a colorimetric NA inhibition assay. The results of assay showed no NAIs-resistant phenotype for any of the viruses. The R118K mutation was the most recurrent, as it was observed in all subtypes except for N6. IC50 values confirmed the differences in sensitivity to OC or ZA observed in the N1 and N2 groups of NAs. Furthermore, both wild types (WTs) in the N6 and one WT in the N9 subtype were less sensitive to ZA than were genotypically related mutants with R152K and R118K change in the respective subtypes. This may indicate that these and probably even other NAIs resistance-related mutations found in our virus collection were not induced by NAIs residuals in the environment and that the impact of such mutations in an avian influenza could be dependent on subtype, strain and host species. PMID:24558492

Orozovic, Goran; Orozovic, Kanita; Jarhult, Josef D.; Olsen, Bjorn

2014-01-01

302

Hepatitis B virus subgenotype A1: evolutionary relationships between Brazilian, African and Asian isolates.  

PubMed

Brazil is a country of low hepatitis B virus (HBV) endemicity in which the genotype A of HBV (HBV/A) is the most prevalent. The complete nucleotide sequences of 26 HBV/A isolates, originating from eight Brazilian states, were determined. All were adw2. Twenty-three belonged to subgenotype A1 and three to A2. By phylogenetic analysis, it was shown that all the 23 HBV/A1 isolates clustered together with isolates from Bangladesh, India, Japan, Nepal, the Philippines and United Arab Emirates, but not with those of Congo, Kenya, Malawi, Rwanda, South Africa, Tanzania, Uganda and Zimbabwe. Four amino acid residues in the polymerase (His138 in the terminal protein domain, Pro18 and His90 in the spacer, and Ser109 in the reverse transcriptase), and one (Phe17) in the precore region, predominated in Latin American and Asian HBV/A1 isolates, but were rarely encountered in African isolates, with the exception of those from Somalia. Specific variations of two adjacent amino acids in the C-terminal domain of the HBx protein, namely Ala146 and Pro147, were found in all the Brazilian, but rarely in the other HBV/A1 isolates. By Bayesian analysis, the existence of an 'Asian-American' clade within subgenotype A1 was supported by a posterior probability value of 0.996. The close relatedness of the Brazilian, Asian and Somalian isolates suggests that the HBV/A1 strains predominant in Brazil did not originate from the five million slaves who were imported from Central and Western Africa from 1551 to 1840, but rather from the 300-400,000 captives forcibly removed from southeast Africa at the middle of the 19th century. PMID:25122004

Lago, Bárbara V; Mello, Francisco C; Kramvis, Anna; Niel, Christian; Gomes, Selma A

2014-01-01

303

Characterization of Newly Emerging Newcastle Disease Virus Isolates from the People's Republic of China and Taiwan  

PubMed Central

Seven Newcastle disease (ND) virus (NDV) isolates which were recovered from ND outbreaks in chicken and pigeon flocks in China and Taiwan between 1996 and 2000 were genotypically and pathotypically characterized. By phylogenetic analysis of the fusion protein genes, isolates Ch-A7/96, Ch/98-3, Ch/99, Ch/2000, and TW/2000 were placed into two novel subgenotypes, VIIc and VIId. Isolate Ch/98-1 was grouped into subgenotype VIb, while Ch-W6/96 was proven to be a mixture of isolates Ch-A7/96 and Ch/98-1. These isolates were pathotyped as viscerotropic velogenic for Ch/98-3, Ch/99, Ch/2000, and TW/2000; neurotropic velogenic for Ch-A7/96; and mesogenic for Ch/98-1. Three separate, comparative, genetic analyses of the F genes, including genetic distance measurement, phylogenetic tree analysis, and residue substitution analysis, were performed with our isolates and selected NDV strains from GenBank. Results showed that the close genetic similarity provided evidence for the epidemiological linkage between the outbreaks in China and Taiwan and that the 1990s outbreaks in Asia, the Middle East, Africa, and Europe constituted the fourth panzootic of ND. In combination with epidemiological analysis, an evolutionary model of the NDV strains, representative of the direction of transmission within the NDV strains, was proposed, and epidemiology of NDV transmission was evaluated with emphasis on molecular aspects. Finally, a cross-protective experiment indicated that at least one strain (Ch-A7/96) among our NDV isolates was an antigenic variant, responsible for recent outbreaks of ND in vaccinated chicken flocks. PMID:11574565

Yu, Li; Wang, Zhiliang; Jiang, Yihai; Chang, Leo; Kwang, Jimmy

2001-01-01

304

Molecular characterization of an infectious bronchitis virus strain isolated from northern China in 2012.  

PubMed

This study reports the complete genome sequence of an infectious bronchitis virus (CK/CH/SD/121220, KJ128295) isolated in 2012 from Shandong Province in northern China. The genome is 27,666 nt long, comprising six genes and 5' and 3' untranslated regions. The full-length genome of the CK/CH/SD/121220 isolate had the highest nucleotide sequence identity (96.7 %) to the YX10 strain. Sites of recombination were identified in the genes 1ab, S, 5a, 5b and N, with their putative parental strains belonging to the QX- and YN-type subgroups, which are already circulating in China. Our findings suggest an important role played by recombination in IBV evolution. PMID:25168045

Zhao, Ye; Liu, Xiao-Yu; Cheng, Jin-Long; Wu, Yan-Ping; Zhang, Guo-Zhong

2014-12-01

305

Genotyping hepatitis C virus isolates from Spain, Brazil, China, and Macau by a simplified PCR method.  

PubMed Central

An improved and simplified method of genotyping was developed for classifying hepatitis C virus (HCV) isolates into the five common genotypes, i.e., I/1a, II/1b, III/2a, IV/2b, and V/3a, by PCR with genotype-specific primers deduced from the core gene. Sense and antisense primers, specific for each of the five common genotypes, were designed by comparison of 319 core gene sequences from HCV isolates of various genotypes from genetic groups 1 to 9. In the first round of PCR, a sequence of 433 bp representing nucleotides 319 to 751 was amplified with universal primers. The second round of PCR was performed with respective sense and antisense primers in two separate reactions, one for the amplification of genotypes I/1a and II/1b and the other for the amplification of genotypes III/2a, IV/2b, and V/3a. The specificity of genotyping was confirmed with a panel of 191 serum samples containing HCV isolates whose core gene sequences were known: 110 serum samples infected with HCV of the five common genotypes and 81 serum samples infected with HCV of other genotypes. The use of sense and antisense primers for genotype II/1b (primers 389 and 492) abolished the cross-reaction of the antisense primer for genotype II/1b (primer 133) with some HCV isolates of genotype I/1a found by our original method. The new method was used for genotyping 130 HCV isolates from Spain, 53 from Brazil, 106 from China, and 30 from Macau. A total of 329 bp of the NS5b region (nucleotides 8279 to 8607) of five isolates from Spain and five isolates from Macau which could not be classified as any of the five common HCV genotypes or genotype 2c were sequenced, and the sequences were compared with those of HCV isolates of known genotypes; two isolates from Spain were deduced to be of genotype 4d and one was deduced to be of genotype 1d, while the remaining two isolates from Spain had novel genotypes in genetic group 2; however, all five isolates from Macau were of genotype 6a. PMID:8880482

Holland, P V; Barrera, J M; Ercilla, M G; Yoshida, C F; Wang, Y; de Olim, G A; Betlach, B; Kuramoto, K; Okamoto, H

1996-01-01

306

Influence of isolate pathogenicity on the aerosol transmission of Porcine reproductive and respiratory syndrome virus  

PubMed Central

The objectives of this study were to evaluate the role of isolate pathogenicity in the aerosol transmission of Porcine reproductive and respiratory syndrome virus (PRRSV) and to determine whether PRRSV could be detected in air samples. To assess transmission, we exposed naďve recipient pigs to aerosols from pigs inoculated with PRRSV MN-30100, an isolate of low pathogenicity, or MN-184, a highly pathogenic isolate. Blood samples and nasal-swab samples were collected from the inoculated pigs during the exposure period and tested for the presence of PRRSV RNA by quantitative (real-time) reverse-transcriptase polymerase chain reaction (RT-PCR); the amount of RNA was expressed as the median tissue culture dose per milliliter (TCID50/mL). The recipient pigs were clinically evaluated for 14 d after exposure and tested on days 7 and 14 by qualitative RT-PCR and enzyme-linked immunosorbent assay (ELISA). To prove the presence of PRRSV in aerosols, air samples were collected from each recipient-pig chamber by means of an air sampler. The PRRSV RNA concentrations were significantly higher (P = 0.01) in the blood samples from the pigs infected with PRRSV MN-184 than in the blood samples from those infected with PRRSV MN-30100; however, the concentrations in the nasal-swab samples were not significantly different (P = 0.26). Recipient pigs exposed to aerosols from pigs infected with PRRSV MN-184 became infected, whereas those exposed to aerosols from pigs infected with PRRSV MN-30100 did not; the difference in transmission rate was significant at P = 0.04. We detected PRRSV MN-184 RNA but not PRRSV MN-30100 RNA in air samples by PCR. Under the conditions of this study, PRRSV isolate pathogenicity may influence aerosol transmission of the virus. PMID:17193878

Cho, Jenny G.; Deen, John; Dee, Scott A.

2007-01-01

307

Isolation of Buggy Creek Virus (Togaviridae: Alphavirus) From Field-Collected Eggs of Oeciacus vicarius (Hemiptera: Cimicidae)  

PubMed Central

Alphaviruses (Togaviridae) rarely have been found to be vertically transmitted from female arthropods to their progeny. We report two isolations of Buggy Creek virus (BCRV), an ecologically unusual alphavirus related to western equine encephalomyelitis virus, from field-collected eggs of cimicid swallow bugs (Oeciacus vicarius Horvath), the principal vector for BCRV. Ten percent of egg pools were positive for BCRV, and we estimated minimum infection rates to be 1.03 infected eggs per 1,000 tested. The results show potential vertical transmission of BCRV, represent one of the few isolations of any alphavirus from eggs or larvae of insects in the field, and are the first report of any virus in the eggs of cimicid bedbugs. The specialized ecological niche of BCRV in swallow bugs and at cliff swallow (Petrochelidon pyrrhonota Vieillot) nesting sites may promote vertical transmission of this virus. PMID:19351091

Brown, Charles R.; Moore, Amy T.; Young, Ginger R.; Padhi, Abinash; Komar, Nicholas

2009-01-01

308

Isolation of Buggy Creek virus (Togaviridae: Alphavirus) from field-collected eggs of Oeciacus vicarius (Hemiptera: Cimicidae).  

PubMed

Alphaviruses (Togaviridae) rarely have been found to be vertically transmitted from female arthropods to their progeny. We report two isolations of Buggy Creek virus (BCRV), an ecologically unusual alphavirus related to western equine encephalomyelitis virus, from field-collected eggs of cimicid swallow bugs (Oeciacus vicarius Horvath), the principal vector for BCRV. Ten percent of egg pools were positive for BCRV, and we estimated minimum infection rates to be 1.03 infected eggs per 1,000 tested. The results show potential vertical transmission of BCRV, represent one of the few isolations of any alphavirus from eggs or larvae of insects in the field, and are the first report of any virus in the eggs of cimicid bedbugs. The specialized ecological niche of BCRV in swallow bugs and at cliff swallow (Petrochelidon pyrrhonota Vieillot) nesting sites may promote vertical transmission of this virus. PMID:19351091

Brown, Charles R; Moore, Amy T; Young, Ginger R; Padhi, Abinash; Komar, Nicholas

2009-03-01

309

Characterization of an Avian Influenza Virus H9N2 Strain Isolated from a Wild Bird in Southern China  

PubMed Central

We isolated an avian influenza virus H9N2 strain from a wild bird in the Guangxi Province of southern China in 2013 named A/turtledove/Guangxi/49B6/2013(H9N2) (GX49B6). We aimed to understand the genetic characters of the GX49B6 strain by analyzing the complete genome sequence. The results showed that our isolated strain has features of low pathogenic avian influenza viruses and viruses that infect humans. The discovery of the complete genome sequence of the GX49B6 strain may be helpful to further the understanding of the epidemiology and surveillance of avian influenza viruses in the field. PMID:24948768

Xu, Qian; Xie, Liji; Xie, Zhiqin; Deng, Xianwen; Liu, Jiabo; Luo, Sisi

2014-01-01

310

Characterization of human immunodeficiency virus type 1-specific cytotoxic T lymphocyte clones isolated during acute seroconversion: recognition of autologous virus sequences within a conserved immunodominant epitope  

PubMed Central

Virus-specific cytotoxic T lymphocytes (CTL) are involved in protective immunity to many virus infections. It has recently been shown that CTL are detectable early during primary infection with the primate lentiviruses, human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus. To better characterize the CTL response during acute HIV-1 infection, HIV-1-specific CTL clones were generated from two patients during symptomatic HIV-1 seroconversion. These CTL clones demonstrated specificity for env of HIV-1 and recognized sequences within gp41. Two human histocompatibility leukocyte antigen (HLA) A31- restricted clones from the same individual were found to have differing virus strain specificities. Both clones recognized the 11-amino acid peptide RLRDLLLIVTR from position 770-780 of gp41. A change from T to V at position 779 in this epitope abrogated lysis by one clone but not the other. A CTL clone from the other patient, restricted by a different class I HLA allele, recognized the nine-amino acid peptide HRLRDLLLI from position 769-777 of gp41. Of note, the peptide RLRDLLLIVTR has been shown by others to be presented to CTL by HLA- A3.1. Autologous virus sequences from seroconversion and up to 15 wk after presentation in these two patients were recognized by the CTL clones isolated during acute infection. None of the CTL clones recognized the MN strain of HIV-1, indicating the problems inherent in relying on a single virus strain in the development of a vaccine. These studies have identified an immunodominant and promiscuous area for the generation of CTL responses within gp41. This recognition of autologous virus sequences by the initial CTL response is consistent with the hypothesis that a single virus strain is transmitted to the seroconverter and that the CTL response is involved in the initial control of that virus. These studies indicate the importance of the CTL response to HIV-1 infection and have implications in the design of vaccines. PMID:8294860

1994-01-01

311

Microevolution of Highly Pathogenic Avian Influenza A(H5N1) Viruses Isolated from Humans, Egypt, 2007–2011  

PubMed Central

We analyzed highly pathogenic avian influenza A(H5N1) viruses isolated from humans infected in Egypt during 2007–2011. All analyzed viruses evolved from the lineage of subtype H5N1 viruses introduced into Egypt in 2006; we found minimal evidence of reassortment and no exotic introductions. The hemagglutinin genes of the viruses from 2011 formed a monophyletic group within clade 2.2.1 that also included human viruses from 2009 and 2010 and contemporary viruses from poultry; this finding is consistent with zoonotic transmission. Although molecular markers suggestive of decreased susceptibility to antiviral drugs were detected sporadically in the neuraminidase and matrix 2 proteins, functional neuraminidase inhibition assays did not identify resistant viruses. No other mutations suggesting a change in the threat to public health were detected in the viral proteomes. However, a comparison of representative subtype H5N1 viruses from 2011 with older subtype H5N1 viruses from Egypt revealed substantial antigenic drift. PMID:23260983

Younan, Mary; Poh, Mee Kian; Elassal, Emad; Davis, Todd; Rivailler, Pierre; Balish, Amanda L.; Simpson, Natosha; Jones, Joyce; Deyde, Varough; Loughlin, Rosette; Perry, Ije; Gubareva, Larisa; ElBadry, Maha A.; Truelove, Shaun; Gaynor, Anne M.; Mohareb, Emad; Amin, Magdy; Cornelius, Claire; Pimentel, Guillermo; Earhart, Kenneth; Naguib, Amel; Abdelghani, Ahmed S.; Refaey, Samir; Klimov, Alexander I.; Kandeel, Amr

2013-01-01

312

Use of lambdagt11 to isolate genes for two pseudorabies virus glycoproteins with homology to herpes simplex virus and varicella-zoster virus glycoproteins  

SciTech Connect

A library of pseudorabies virus (PRV) DNA fragments was constructed in the expression cloning vector lambdagt11. The library was screened with antisera which reacted with mixtures of PRV proteins to isolate recombinant bacteriophages expressing PRV proteins. By the nature of the lambdagt11 vector, the cloned proteins were expressed in Escherichia coli as ..beta..-galactosidase fusion proteins. The fusion proteins from 35 of these phages were purified and injected into mice to raise antisera. The antisera were screened by several different assays, including immunoprecipitation of (/sup 14/C)glucosamine-labeled PRV proteins. This method identified phages expressing three different PRV glycoproteins: the secreted glycoprotein, gX; gI; and a glycoprotein that had not been previously identified, which we designate gp63. The gp63 and gI genes map adjacent to each other in the small unique region of the PRV genome. The DNA sequence was determined for the region of the genome encoding gp63 and gI. It was found that gp63 has a region of homology with a herpes simplex virus type 1 (HSV-1) protein, encoded by US7, and also with varicella-zoster virus (VZV) gpIV. The gI protein sequence has a region of homology with HSV-1 gE and VZV gpI. It is concluded that PRV, HSV, and VZV all have a cluster of homologous glycoprotein genes in the small unique components of their genomes and that the organization of these genes is conserved.

Petrovskis, E.A.; Timmins, J.G.; Post, L.E.

1986-10-01

313

Efficacy of a whole inactivated EI vaccine against a recent EIV outbreak isolate and comparative detection of virus shedding  

Microsoft Academic Search

An outbreak of H3N8 Equine Influenza virus (EIV) that occurred in vaccinated horses in Japan was caused by a genetically divergent EIV isolate of the Florida clade 1 sub-lineage. This virus subsequently entered Australia where it infected thousands of immunologically naďve horses. The objective of this study was to evaluate the ability of a non-updated whole inactivated equine influenza (EI)

R. Paillot; L. Prowse; C. Donald; E. Medcalf; F. Montesso; N. Bryant; J. Watson; M. Jeggo; D. Elton; R. Newton; P. Trail; H. Barnes

2010-01-01

314

Some properties of pea enation mosaic virus isolated from field pea and broad bean plants in Bohemia  

Microsoft Academic Search

Pea enation mosaic virus (PEMV) was isolated from disea sed field pea (Pisum sativum L.ssp. arvense A.Gr.) and broad bean (Faba vulgaris Moench) plants grown as filed crops at Bohumilice in Bohemia. The virus proved to be pathogenic for the following plant species:Pisum sativum L. cv. Raman,Faba vulgaris Moench,Lens culinaris Med.,Vicia sativa L.,Lathyrus odoratus L.,Glycine soja L.,Phaseolus vulgaris L.,Chenopodium amaranticolor

M. Musil; Olga Lešková

1969-01-01

315

Isolation and characterization of simian immunodeficiency virus from African white-crowned mangabey monkeys ( Cercocebus torquatus lunulatus )  

Microsoft Academic Search

Summary Forty-eight of 236 sera from seven species of African non-human primates in Kenya, including those of white-crowned mangabey monkeys (Cercocebus torquatus lunulatus) had antibodies to simian immunodeficiency viruses (SIVs). Isolates of simian lentivirus were obtained from seropositive white-crowned mangabey monkeys which are indigenous in West Africa. This virus, designated as SIVWCM, appeared morphologically similar to HIV by electron microscopy,

K. Tomonaga; J. Katahira; M. Fukasawa; M. A. Hassan; M. Kawamura; H. Akari; T. Miura; T. Goto; M. Nakai; M. Suleman; M. Isahakia; M. Hayami

1993-01-01

316

Molecular characterization and phylogenetics of a reassortant H13N8 influenza virus isolated from gulls in Mongolia.  

PubMed

Double reassortant H13N8 influenza A virus was isolated from gull in Mongolia. The basic virological characteristics were studied. Complete genome sequence analysis indicated the complicated evolutionary history. The PA gene belongs to classical Avian-like lineage and more likely originated from non-gull avian virus pool. Data confirm the state of extensive geographic mosaicism in AIV from gulls in the Northern Hemisphere. PMID:24839173

Sharshov, K; Sivay, M; Liu, D; Pantin-Jackwood, M; Marchenko, V; Durymanov, A; Alekseev, A; Damdindorj, T; Gao, G F; Swayne, D E; Shestopalov, A

2014-10-01

317

Molecular epidemiology of Nigerian and Ghanaian measles virus isolates reveals a genotype circulating widely in western and central Africa  

Microsoft Academic Search

Sub-Saharan Africa is one of the regions of the globe with the highest measles-related morbidity and mortality. Yet only seven virus isolates from this vast region have been phylogenetically characterized on the basis of their nucleoprotein, the last one in 1991. To characterize the prevalent wild-type viruses and to understand their circulation pattern, a large panel (n fl 45) of

Frank Hanses; Anh T. Truong; Wim Ammerlaan; Oyewole Ikusika; Festus Adu; Akeeb O. Oyefolu; Sunday A. Omilabu; Claude P. Muller

1999-01-01

318

Hepatitis B virus genotypes circulating in Brazil: molecular characterization of genotype F isolates  

PubMed Central

Background Hepatitis B virus (HBV) isolates have been classified in eight genotypes, A to H, which exhibit distinct geographical distributions. Genotypes A, D and F are predominant in Brazil, a country formed by a miscegenated population, where the proportion of individuals from Caucasian, Amerindian and African origins varies by region. Genotype F, which is the most divergent, is considered indigenous to the Americas. A systematic molecular characterization of HBV isolates from different parts of the world would be invaluable in establishing HBV evolutionary origins and dispersion patterns. A large-scale study is needed to map the region-by-region distribution of the HBV genotypes in Brazil. Results Genotyping by PCR-RFLP of 303 HBV isolates from HBsAg-positive blood donors showed that at least two of the three genotypes, A, D, and F, co-circulate in each of the five geographic regions of Brazil. No other genotypes were identified. Overall, genotype A was most prevalent (48.5%), and most of these isolates were classified as subgenotype A1 (138/153; 90.2%). Genotype D was the most common genotype in the South (84.2%) and Central (47.6%) regions. The prevalence of genotype F was low (13%) countrywide. Nucleotide sequencing of the S gene and a phylogenetic analysis of 32 HBV genotype F isolates showed that a great majority (28/32; 87.5%) belonged to subgenotype F2, cluster II. The deduced serotype of 31 of 32 F isolates was adw4. The remaining isolate showed a leucine-to-isoleucine substitution at position 127. Conclusion The presence of genotypes A, D and F, and the absence of other genotypes in a large cohort of HBV infected individuals may reflect the ethnic origins of the Brazilian population. The high prevalence of isolates from subgenotype A1 (of African origin) indicates that the African influx during the colonial slavery period had a major impact on the circulation of HBV genotype A currently found in Brazil. Although most genotype F isolates belonged to cluster II, the presence of some isolates belonging to clusters I (subgroup Ib) and IV suggests the existence of two or more founder viral populations of genotype F in Brazil. PMID:18036224

Mello, Francisco CA; Souto, Francisco JD; Nabuco, Leticia C; Villela-Nogueira, Cristiane A; Coelho, Henrique Sergio M; Franz, Helena Cristina F; Saraiva, Joao Carlos P; Virgolino, Helaine A; Motta-Castro, Ana Rita C; Melo, Mabel MM; Martins, Regina MB; Gomes, Selma A

2007-01-01

319

Rapid detection of infectious laryngotracheitis virus isolates by loop-mediated isothermal amplification.  

PubMed

The objective of this study was to develop and evaluate a loop-mediated isothermal amplification (LAMP) method to detect infectious laryngotracheitis virus (ILTV) from commercial broiler and layer flocks in southern China. A set of six specific primers was designed to recognize six distinct genomic sequences of thymidine kinase (TK) from ILTV. The entire assay duration was recorded at 40 min under isothermal condition at 63.5 degrees C. The amplified products were analyzed by electrophoresis and visual judgment by the SYBR Green I dyeing. LAMP assay was 10-fold more sensitive than the routine PCR assay, with a detection limit of 46 copies per reaction. In detecting ILTV, the LAMP assay detected all 5 strains previously isolated, did not cross-react with other avian pathogens, and obtained a 100% sensitivity in 43 positive clinical samples with reference to virus isolation. Therefore, the LAMP assay may be a good alternative method for specific diagnosis of ILTV infection in primary care facilities, and in less well-equipped laboratories. PMID:20100518

Xie, Qing-mei; Ji, Jun; Pickens, Tristan Tyler; Du, Li-qin; Cao, Yong-chang; Li, Hong-mei; Wang, Lin-guo; Ma, Jing-yun; Bi, Ying-zuo

2010-04-01

320

The genome sequence of ectromelia virus Naval and Cornell isolates from outbreaks in North America.  

PubMed

Ectromelia virus (ECTV) is the causative agent of mousepox, a disease of laboratory mouse colonies and an excellent model for human smallpox. We report the genome sequence of two isolates from outbreaks in laboratory mouse colonies in the USA in 1995 and 1999: ECTV-Naval and ECTV-Cornell, respectively. The genome of ECTV-Naval and ECTV-Cornell was sequenced by the 454-Roche technology. The ECTV-Naval genome was also sequenced by the Sanger and Illumina technologies in order to evaluate these technologies for poxvirus genome sequencing. Genomic comparisons revealed that ECTV-Naval and ECTV-Cornell correspond to the same virus isolated from independent outbreaks. Both ECTV-Naval and ECTV-Cornell are extremely virulent in susceptible BALB/c mice, similar to ECTV-Moscow. This is consistent with the ECTV-Naval genome sharing 98.2% DNA sequence identity with that of ECTV-Moscow, and indicates that the genetic differences with ECTV-Moscow do not affect the virulence of ECTV-Naval in the mousepox model of footpad infection. PMID:24999046

Mavian, Carla; López-Bueno, Alberto; Bryant, Neil A; Seeger, Kathy; Quail, Michael A; Harris, David; Barrell, Bart; Alcami, Antonio

2014-08-01

321

Epidemiology and genetic characterization of equine infectious anaemia virus strains isolated in belgium in 2010.  

PubMed

In January 2010, the United Kingdom notified cases of equine infectious anaemia (EIA) in two horses introduced from Belgium. The animals came from one assembly centre in Romania and had transited through Belgium with 16 other horses. Nine of them, bought by a Belgian horse breeder, were investigated in Belgium and revealed one additional EIA-positive animal. Afterwards, the Belgian Federal Agency for the Safety of the Food Chain (FASFC) organized a serological EIA survey of the horses introduced into Belgium from Romania between 2007 and 2009. Among the 95 horses identified, six additional serological positive cases were found that had been introduced into Belgium in 2008 (n = 4) and in 2009 (n = 2). The survey was extended to the horses in contact with the positive cases, but all contact animals were negative, indicating the absence of transmission. Virological examination performed on tissue samples collected from two seropositive animals demonstrated the presence of viral DNA of EIA virus. Phylogenetic analysis based on the sequences of EIA virus gag gene clustered the Belgian isolates with Romanian strains isolated in 2009. The presumption of a common Belgian origin could be rejected. PMID:23173784

Caij, A B; Tignon, M

2014-10-01

322

The genome sequence of ectromelia virus Naval and Cornell isolates from outbreaks in North America  

PubMed Central

Ectromelia virus (ECTV) is the causative agent of mousepox, a disease of laboratory mouse colonies and an excellent model for human smallpox. We report the genome sequence of two isolates from outbreaks in laboratory mouse colonies in the USA in 1995 and 1999: ECTV-Naval and ECTV-Cornell, respectively. The genome of ECTV-Naval and ECTV-Cornell was sequenced by the 454-Roche technology. The ECTV-Naval genome was also sequenced by the Sanger and Illumina technologies in order to evaluate these technologies for poxvirus genome sequencing. Genomic comparisons revealed that ECTV-Naval and ECTV-Cornell correspond to the same virus isolated from independent outbreaks. Both ECTV-Naval and ECTV-Cornell are extremely virulent in susceptible BALB/c mice, similar to ECTV-Moscow. This is consistent with the ECTV-Naval genome sharing 98.2% DNA sequence identity with that of ECTV-Moscow, and indicates that the genetic differences with ECTV-Moscow do not affect the virulence of ECTV-Naval in the mousepox model of footpad infection. PMID:24999046

Mavian, Carla; Lopez-Bueno, Alberto; Bryant, Neil A.; Seeger, Kathy; Quail, Michael A.; Harris, David; Barrell, Bart; Alcami, Antonio

2014-01-01

323

Screening, isolation and optimization of anti-white spot syndrome virus drug derived from marine plants  

PubMed Central

Objective To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various marine floral ecosystems and to evaluate the efficacy of the same in host–pathogen interaction model. Methods Thirty species of marine plants were subjected to Soxhlet extraction using water, ethanol, methanol and hexane as solvents. The 120 plant isolates thus obtained were screened for their in vivo anti-WSSV property in Litopenaeus vannamei. By means of chemical processes, the purified anti-WSSV plant isolate, MP07X was derived. The drug was optimized at various concentrations. Viral and immune genes were analysed using reverse transcriptase PCR to confirm the potency of the drug. Results Nine plant isolates exhibited significant survivability in host. The drug MP07X thus formulated showing 85% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of MP07X required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 1?000 mg/kg body weight/day survived at the rate of 85%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection. Conclusions The drug MP07X derived from Rhizophora mucronata is a potent anti-WSSV drug. PMID:25183065

Chakraborty, Somnath; Ghosh, Upasana; Balasubramanian, Thangavel; Das, Punyabrata

2014-01-01

324

A comparative study of strains of Ebola virus isolated from southern Sudan and northern Zaire in 1976.  

PubMed

During the 1976 Ebola virus outbreak in Sudan, the investigations team gained the impression that fewer haemorrhagic manifestations and few fatalities occurred during the later stages of the epidemic after the virus had undergone several generations in man. This impression was also noted in guinea pigs experimentally infected with Sudanese and Zairean strains of Ebola virus. The virulence of the Sudanese isolates was less intense than isolates emanating from Zaire. Similar findings were seen in monkeys; a Zairean isolated produced fatal infections, whereas monkeys inoculated with a Sudan strain generally recovered. Two monkeys, which had recovered from Sudanese strain infections and had developed high levels of antibody detectable by immunofluorescence, were challenged with the Zairean strain. Both developed viraemias and died. The mechanisms of this "failed protection" are discussed. PMID:6165800

Bowen, E T; Platt, G S; Lloyd, G; Raymond, R T; Simpson, D I

1980-01-01

325

Detection and Isolation of Airborne Influenza A H3N2 Virus Using a Sioutas Personal Cascade Impactor Sampler  

PubMed Central

The air we breathe contains microorganisms that can cause infectious respiratory diseases. After two occupants of an apartment were diagnosed with influenza in February of 2013, efforts were made to detect and isolate airborne influenza virus using two different types of active air samplers: a Sioutas Personal Cascade Impactor Sampler (PCIS) and an SKC BioSampler. The PCIS collects size-fractionated particles by impaction on polytetrafluoroethylene filters, whereas the SKC BioSampler collects airborne particles in liquid media. Influenza H3N2 virus was collected by both types of air samplers. The PCIS collected a range of particle sizes containing influenza virus near one of the sick individuals but only ultrafine particles when the samplers were positioned farther away. Viable virus was present in the liquid collection media of the SKC BioSampler and some PCIS filters. These findings suggest that influenza patients produce ultrafine aerosol particles that contain viable virus. PMID:24224087

Lednicky, John A.; Loeb, Julia C.

2013-01-01

326

Detection and Isolation of Airborne Influenza A H3N2 Virus Using a Sioutas Personal Cascade Impactor Sampler.  

PubMed

The air we breathe contains microorganisms that can cause infectious respiratory diseases. After two occupants of an apartment were diagnosed with influenza in February of 2013, efforts were made to detect and isolate airborne influenza virus using two different types of active air samplers: a Sioutas Personal Cascade Impactor Sampler (PCIS) and an SKC BioSampler. The PCIS collects size-fractionated particles by impaction on polytetrafluoroethylene filters, whereas the SKC BioSampler collects airborne particles in liquid media. Influenza H3N2 virus was collected by both types of air samplers. The PCIS collected a range of particle sizes containing influenza virus near one of the sick individuals but only ultrafine particles when the samplers were positioned farther away. Viable virus was present in the liquid collection media of the SKC BioSampler and some PCIS filters. These findings suggest that influenza patients produce ultrafine aerosol particles that contain viable virus. PMID:24224087

Lednicky, John A; Loeb, Julia C

2013-01-01

327

Characterization of a herpes virus isolated from domestic geese in Australia  

USGS Publications Warehouse

A herpesvirus (GHV 552/89) associated with high mortality in a flock of domestic geese in Australia was compared with duck virus enteritis (DVE) herpesvirus by cross-protection studies in domestic geese, Muscovy ducks and commercial Pekin ducks. In DVE-vaccinated geese, Muscovy ducks and Pekin ducks, mortality levels of 100, 50 and 0%, respectively, were recorded following challenge with GHV 552/89. Conversely, in geese, Muscovy ducks and Pekin ducks immunized with inactivated GHV 552/89, 100% mortality was observed in the geese and Muscovy ducks, and 80% in the Pekin ducks following challenge with DVE virus. The isolate was also compared with six other avian herpesviruses using cross-neutralization tests in cell cultures. No detectable cross-neutralization occurred with any of the avian herpesviruses tested. Further characterization of GHV 552/89 was undertaken by comparing its genome with strains of DVE herpesvirus using restriction endonuclease analysis of the viral DNA and a polymerase chain reaction (PCR) test. Following digestion with HindIII, the DNA fragment pattern of GHV 552/89 was found to be completely different from the DVE viruses. Similarities were found between the digestion patterns of a UK and a US DVE isolate, but both were distinguishable from a UK vaccine strain. The results of the PCR analysis and comparison using two DVE-specific primer sets did not produce specific amplification products of expected molecular weights (603 and 446 base pairs) from the GHV 552/89 genome. The PCR products derived from the DVE strains were similar to those derived from the DVE control DNA. From the results of this study, it is concluded that the goose herpesvirus GHV 552/89 is antigenically and genomically distinct from DVE herpesvirus.

Gough, R.E.; Hansen, W.R.

2000-01-01

328

Complete Genome Sequence of an H10N5 Avian Influenza Virus Isolated from Pigs in Central China  

PubMed Central

An avian H10N5 influenza virus, A/swine/Hubei/10/2008/H10N5, was isolated from pigs in the Hubei Province of central China. Homology and phylogenetic analyses of all eight gene segments demonstrated that the strain was wholly of avian origin and closely homologous to the Eurasian lineage avian influenza virus. To our knowledge, this is the first report of interspecies transmission of an avian H10N5 influenza virus to domestic pigs under natural conditions. PMID:23166264

Wang, Nan; Zou, Wei; Yang, Ying; Guo, Xuebo; Hua, Yafeng; Zhang, Qiang; Zhao, Zongzheng

2012-01-01

329

Pathogenicity and Transmission in Pigs of the Novel A(H3N2)v Influenza Virus Isolated from Humans and Characterization of Swine H3N2 Viruses Isolated in 2010-2011  

PubMed Central

Swine influenza virus (SIV) H3N2 with triple reassorted internal genes (TRIG) has been enzootic in Unites States since 1998. Transmission of the 2009 pandemic H1N1 (pH1N1) virus to pigs in the United States was followed by reassortment with endemic SIV, resulting in reassorted viruses that include novel H3N2 genotypes (rH3N2p). Between July and December 2011, 12 cases of human infections with swine-lineage H3N2 viruses containing the pandemic matrix (pM) gene [A(H3N2)v] were detected. Whole-genome analysis of H3N2 viruses isolated from pigs from 2009 to 2011 sequenced in this study and other available H3N2 sequences showed six different rH3N2p genotypes present in the U.S. swine population since 2009. The presence of the pM gene was a common feature among all rH3N2p genotypes, but no specific genotype appeared to predominate in the swine population. We compared the pathogenic, transmission, genetic, and antigenic properties of a human A(H3N2)v isolate and two swine H3N2 isolates, H3N2-TRIG and rH3N2p. Our in vivo study detected no increased virulence in A(H3N2)v or rH3N2p viruses compared to endemic H3N2-TRIG virus. Antibodies to cluster IV H3N2-TRIG and rH3N2p viruses had reduced cross-reactivity to A(H3N2)v compared to other cluster IV H3N2-TRIG and rH3N2p viruses. Genetic analysis of the hemagglutinin gene indicated that although rH3N2p and A(H3N2)v are related to cluster IV of H3N2-TRIG, some recent rH3N2p isolates appeared to be forming a separate cluster along with the human isolates of A(H3N2)v. Continued monitoring of these H3N2 viruses is necessary to evaluate the evolution and potential loss of population immunity in swine and humans. PMID:22491461

Kitikoon, Pravina; Gauger, Phillip C.; Schlink, Sarah N.; Bayles, Darrell O.; Gramer, Marie R.; Darnell, Daniel; Webby, Richard J.; Lager, Kelly M.; Swenson, Sabrina L.; Klimov, Alexander

2012-01-01

330

Isolation and Characterization of a Single-Stranded RNA Virus Infecting the Bloom-Forming Diatom Chaetoceros socialis?  

PubMed Central

Diatoms are very significant primary producers in the world's oceans. Various environmental factors affect the depletion of diatom populations. The importance of viruses as a potential mortality source has recently been recognized. We isolated and characterized a new diatom virus (Chaetoceros socialis f. radians RNA virus [CsfrRNAV]) causing the lysis of the bloom-forming species Chaetoceros socialis Lauder f. radians (Schütt) Proschkina-Lavrenko. The virus infectious to C. socialis f. radians was isolated from water samples collected in Hiroshima Bay. Here we show the physiology, morphology, and genome characteristics of the virus clone. Virions were 22 nm in diameter and accumulated in the cytoplasm of the host cells. The latent period and the burst size were estimated to be <48 h and 66 infectious units per host cell, respectively. CsfrRNAV harbors a single-stranded RNA (ssRNA) genome and encodes at least three polypeptides of 32.0, 28.5, and 25.0 kDa. Sequencing analysis shows the length of the genome is 9,467 bases, excluding a poly(A) tail. The monophyly of CsfrRNAV and other diatom-infecting RNA viruses, Rhizosolenia setigera RNA virus and Chaetoceros tenuissimus RNA virus, was strongly supported by phylogenetic analysis based on the amino acid sequence of the RNA-dependent RNA polymerase domains. This suggested a new ssRNA virus family, Bacillariornaviridae. This discovery of CsfrRNAV may aid in further understanding the ecological dynamics of the C. socialis f. radians population in nature and the relationships between ssRNA diatom viruses and their hosts. PMID:19233955

Tomaru, Yuji; Takao, Yoshitake; Suzuki, Hidekazu; Nagumo, Tamotsu; Nagasaki, Keizo

2009-01-01

331

Problems in the Laboratory Isolation of Simian Hemorrhagic Fever Viruses and Isolation of the Agent Responsible for the Sussex/69 Epizootic  

PubMed Central

At least six epizootics of simian hemorrhagic fever have occurred at four different primate centers. Although these diseases could easily be transmitted to other monkeys of the Macaca species, difficulty has been encountered in isolating the causative virus in cell culture. The results of this study have shown that the isolation of simian hemorrhagic fever virus strains in cell culture is dependent upon the use of a susceptible MA-104 cell strain and that the ability of such strains to support the replication of these viral agents may vary. By using this information we have been able to isolate a viral agent in cell culture from materials derived from the Sussex/69 epizootic. PMID:4626908

Myers, Martin G.; Vincent, Monroe M.; Hensen, Sally A.; Tauraso, Nicola M.

1972-01-01

332

Survival of avian influenza and Newcastle disease viruses in compost and at ambient temperatures based on virus isolation and real-time reverse transcriptase PCR.  

PubMed

In four composting experiments, survival of avian influenza (AI) and Newcastle disease (ND) viruses was assessed by virus isolation in embryonated chicken eggs (ECEs) and by real-time reverse transcriptase-polymerase chain reaction. Specimens contained in nylon mesh bags consisted of 20-g samples of chicken manure, used litter, or feed that had been inoculated with allantoic fluid containing an AI virus (H6N2, Expt. 1) or an ND vaccine virus (Expt. 2). Other specimens consisted of 20-g samples of infected ECEs that had been homogenized and mixed with corn silage. As a control, allantoic fluid diluted in phosphate-buffered saline was contained in sealed vials. Except for the feed, in which the AI virus was inactivated soon after the specimen was inoculated, on day 0 the specimens buried in compost or placed outside at ambient temperatures contained at least 5.0 log10 of virus and 7.7 log10 of viral RNA. By day 7, temperatures in compost ranged from 50 C to 65 C, and viruses had been killed in all specimens in bags. In comparison, viruses in sealed vials remained viable to day 10. Viral RNA in mesh-bag specimens had been degraded to nondetectable levels by day 10, but it was still detected in sealed vials on day 21. In specimens that were held at ambient temperatures (13 C-28 C), the viruses in mesh-bag specimens were inactivated by day 21, but their RNA was still detected. In comparison, the viruses in sealed vials survived to day 21. In Expts. 3 and 4, viruses were inactivated in carcass specimens and in whole ECEs during composting. In an in vitro experiment, the time required for a 1-log10 reduction of viruses was significantly shorter (P < 0.05) in water extracts from compost than in phosphate buffers at temperatures of 25 C to 45 C. This study provided evidence that microbial activity during composting contributed to the rapid killing of AI and ND viruses and to the degradation of their viral RNA. PMID:19432000

Guan, J; Chan, M; Grenier, C; Wilkie, D C; Brooks, B W; Spencer, J L

2009-03-01

333

Restriction fragment length polymorphism analysis of multiple genome regions of Korean isolates of infectious laryngotracheitis virus collected from chickens.  

PubMed

This study was conducted to characterize infectious laryngotracheitis (ILT) viruses isolated from poultry in South Korea using RFLP analysis of PCR products. Seven wild-type Korean isolates from commercial chicken farms collected between 1986 and 2012 were compared with 3 imported commercial vaccine strains [LT Blen (Hudson strain, United States), Laryngo Vac (Cover strain, United States), and Nobilis ILT (Serva strain, France)] and a Korean chicken embryo origin (CEO) vaccine strain [ILT-VAC (Gyeonggi97 strain, Korea)]. Six of the field isolates were highly virulent viruses, and the Kr12AD37 isolate was considered an attenuated type according to Han's RFLP method. These virulent Korean ILT viruses were divided into 3 classes (class I, II, and III). The Kr12AD37 isolate was found to have the same RFLP pattern as the Korean CEO vaccine strain, and both of these strains were different from the 3 foreign vaccine strains. The results suggest that the Korean CEO vaccine strain has been responsible for recent outbreaks, and the characterization of ILT viruses by RFLP was useful for diagnosis by providing epidemiological information. PMID:23873552

Kim, Hye-Ryoung; Kang, Min-Su; Kim, Mi-Jin; Lee, Hee-Soo; Kwon, Yong-Kuk

2013-08-01

334

Stone Lakes Virus (Family Togaviridae, Genus Alphavirus), a Variant of Fort Morgan Virus Isolated From Swallow Bugs (Hemiptera: Cimicidae) West of the Continental Divide  

PubMed Central

Multiple isolates of an alphaviruses within the western equine encephalomyelitis-serocomplex that were related closely to Ft. Morgan and its variant Buggy Creek virus were made from swallow bugs, Oeciacus vicarius Horvath (Hemiptera: Cimicidae), collected from cliff swallow (Petrochelidon pyrrhonota) nests at the Stone Lakes National Wildlife Refuge, Sacramento County, CA, during the summers of 2005 and 2006. This virus (hereafter Stone Lakes virus, family Togaviridae, genus Alphavirus, STLV) was the first record of this viral group west of the Continental Divide. STLV replicated well in Vero and other vertebrate cell cultures but failed to replicate in C6/36 cells or infect Culex tarsalis Coquillett mosquitoes. STLV failed to produce elevated viremias in adult chickens or house sparrows and was weakly immunogenic. In addition, STLV was not isolated from cliff swallow nestlings nor was antibody detected in adults collected at mist nets. We suggest that STL and related swallow bug viruses may be primarily infections of cimicids that are maintained and amplified either by vertical or nonviremic transmission and that cliff swallows may primarily be important as a bloodmeal source for the bugs rather than as an amplification host for the viruses. PMID:19769055

Brault, Aaron C.; Armijos, M. Veronica; Wheeler, Sarah; Wright, Stan; Fang, Ying; Langevin, Stanley; Reisen, William K.

2009-01-01

335

Stone Lakes virus (family Togaviridae, genus Alphavirus), a variant of Fort Morgan virus isolated from swallow bugs (Hemiptera: Cimicidae) west of the Continental Divide.  

PubMed

Multiple isolates of an alphaviruses within the western equine encephalomyelitis-serocomplex that were related closely to Ft. Morgan and its variant Buggy Creek virus were made from swallow bugs, Oeciacus vicarius Horvath (Hemiptera: Cimicidae), collected from cliff swallow (Petrochelidon pyrrhonota) nests at the Stone Lakes National Wildlife Refuge, Sacramento County, CA, during the summers of 2005 and 2006. This virus (hereafter Stone Lakes virus, family Togaviridae, genus Alphavirus, STLV) was the first record of this viral group west of the Continental Divide. STLV replicated well in Vero and other vertebrate cell cultures but failed to replicate in C6/36 cells or infect Culex tarsalis Coquillett mosquitoes. STLV failed to produce elevated viremias in adult chickens or house sparrows and was weakly immunogenic. In addition, STLV was not isolated from cliff swallow nestlings nor was antibody detected in adults collected at mist nets. We suggest that STL and related swallow bug viruses may be primarily infections of cimicids that are maintained and amplified either by vertical or nonviremic transmission and that cliff swallows may primarily be important as a bloodmeal source for the bugs rather than as an amplification host for the viruses. PMID:19769055

Brault, Aaron C; Armijos, M Veronica; Wheeler, Sarah; Wright, Stan; Fang, Ying; Langevin, Stanley; Reisen, William K

2009-09-01

336

Full-Genome Sequence Analysis of a Natural Reassortant H4N2 Avian Influenza Virus Isolated from a Domestic Duck in Southern China  

PubMed Central

We report here the complete genome sequence of a novel reassortant H4N2 avian influenza virus strain, A/duck/Guangxi/125D17/2012(H4N2) (GX125D17), isolated from a duck in Guangxi Province, China in 2012. We obtained the complete genome sequence of the GX125D17 virus isolation by PCR, cloning, and sequencing. Sequence analysis revealed that this H4N2 virus strain was a novel reassortant avian in?uenza virus (AIV). Information about the complete genome sequence of the GX125D17 virus strain will be useful for epidemiological studies. PMID:25212619

Wu, Aiqiong; Xie, Liji; Xie, Zhiqin; Luo, Sisi; Deng, Xianwen; Huang, Li; Huang, Jiaoling; Zeng, Tingting

2014-01-01

337

Antibody-dependent cellular cytotoxicity detects type- and strain-specific antigens among human immunodeficiency virus types 1 and 2 and simian immunodeficiency virus SIVmac isolates.  

PubMed Central

Human cell lines were infected with different strains of human immunodeficiency virus types 1 and 2 (HIV-1 and HIV-2) as well as with a simian immunodeficiency virus SIVmac isolate and used as targets in an antibody-dependent cellular cytotoxicity (ADCC) assay. Sera from HIV-1- or HIV-2-infected subjects provided the antibody, and lymphocytes from normal donors provided the effector cells. About 60% of HIV-1 antibody-positive sera mediated ADCC when tested against any given HIV-1 isolate-infected target cell (human T-cell lymphotropic virus type IIIB, B40, A2587), and about 75% of HIV-2 antibody-positive sera mediated ADCC when tested against target cells infected with HIV-2 isolates (lymphadenopathy-associated virus type 2 and SBL-6669) or simian immunodeficiency virus from macaques. Within each type, individual sera showed different reactivity patterns, and the probability that a serum was ADCC positive was higher when it was tested against several strains. When the ADCC reactivity of sera against different strains was compared, diversity as detected by ADCC appeared to be greater among HIV-1 strains than among HIV-2 strains. For HIV-1, 54 to 67% of the sera gave concordant ADCC reactions, whereas for HIV-2 and SIVmac, 91% of the sera gave concordant results. Almost no strain-specific differences were seen between SBL-6669 and lymphadenopathy-associated virus type 2. As we determined previously, HIV-1 and HIV-2 did not cross-react in ADCC. The results indicated that HIV-1 and HIV-2 antibody-positive sera mediate both strain- and type-specific ADCC. HIV-2 antibody-positive sera seem to mediate ADCC with broader reactivity and to a higher frequency compared with HIV-1 antibody-positive sera. PMID:2746734

Ljunggren, K; Biberfeld, G; Jondal, M; Fenyo, E M

1989-01-01

338

Isolation of viral haemorrhagic septicaemia virus from muskellunge, Esox masquinongy (Mitchill), in Lake St Clair, Michigan, USA reveals a new sublineage of the North American genotype  

Microsoft Academic Search

Viral haemorrhagic septicaemia virus (VHSV) was isolated from muskellunge, Esox masquinongy (Mitchill), caught from the NW portion of Lake St Clair, Michigan, USA in 2003. Affected fish exhibited congestion of internal organs; the inner wall of the swim bladder was thickened and con- tained numerous budding, fluid-filled vesicles. A virus was isolated using fish cell lines inoculated with a homogenate

E Elsayed; M Faisal; M Thomas; G Whelan; W Batts; J Winton

2006-01-01

339

Development of a surveillance scheme for equine influenza in the UK and characterisation of viruses isolated in Europe, Dubai and the USA from 2010-2012.  

PubMed

Equine influenza viruses are a major cause of respiratory disease in horses worldwide and undergo antigenic drift. Several outbreaks of equine influenza occurred worldwide during 2010-2012, including in vaccinated animals, highlighting the importance of surveillance and virus characterisation. Virus isolates were characterised from more than 20 outbreaks over a 3-year period, including strains from the UK, Dubai, Germany and the USA. The haemagglutinin-1 (HA1) sequence of all isolates was determined and compared with OIE-recommended vaccine strains. Viruses from Florida clades 1 and 2 showed continued divergence from each other compared with 2009 isolates. The antigenic inter-relationships among viruses were determined using a haemagglutination-inhibition (HI) assay with ferret antisera and visualised using antigenic cartography. All European isolates belonged to Florida clade 2, all those from the USA belonged to Florida clade 1. Two subpopulations of clade 2 viruses were isolated, with either substitution A144V or I179V. Isolates from Dubai, obtained from horses shipped from Uruguay, belonged to Florida clade 1 and were similar to viruses isolated in the USA the previous year. The neuraminidase (NA) sequence of representative strains from 2007 and 2009 to 2012 was also determined and compared with that of earlier isolates dating back to 1963. Multiple changes were observed at the amino acid level and clear distinctions could be made between viruses belonging to Florida clade 1 and clade 2. PMID:24480583

Woodward, Alana L; Rash, Adam S; Blinman, Donna; Bowman, Samantha; Chambers, Thomas M; Daly, Janet M; Damiani, Armando; Joseph, Sunitha; Lewis, Nicola; McCauley, John W; Medcalf, Liz; Mumford, Jenny; Newton, J Richard; Tiwari, Ashish; Bryant, Neil A; Elton, Debra M

2014-03-14

340

Comparative full length genome sequence analysis of usutu virus isolates from Africa  

PubMed Central

Background Usutu virus (USUV), a flavivirus belonging to the Japanese encephalitis serocomplex, was identified in South Africa in 1959 and reported for the first time in Europe in 2001. To date, full length genome sequences have been available only for the reference strain from South Africa and a single isolate from each of Austria, Hungary, and Italy. Methods We sequenced four USUV isolates from Senegal and the Central African Republic (CAR) between 1974 and 2007 and compared the sequence data to USUV strains from Austria, Hungary, Italy, and South Africa using a Bayesian Markov chain Monte Carlo method. We further clarified the taxonomic status of a USUV strain isolated in CAR in 1969 and proposed earlier as a subtype of USUV due to an asymetric serological cross-reactivity with USUV reference strain. Results A comparison of the four newly obtained USUV sequences with those from SouthAfrica_1959, Vienna_2001, Budapest_2005, and Italy_2009 revealed that they are all 96-99% and 99% similar at the nucleotide and amino acid levels, respectively. The phylogenetic relationships between these sequences indicated that a strain isolated in Senegal in 1993 is most closely related to the USUV strains detected in Europe. Analysis of a strain isolated from a human in CAR in 1981 (CAR_1981) revealed the presence of specific amino acid substitutions and a deletion in the 3? noncoding region. This is the first fully sequenced human USUV isolate. The putative USUV subtype, CAR_1969, was 81% and 94% identical at the nucleotide and amino acid levels, respectively, compared to the other USUV strains. Our phylogenetic analyses support the serological identification of CAR_1969 as a subtype of USUV. Conclusions In this study, we investigate the genetic diversity of USUV in Africa and the phylogenetic relationship of isolates from Africa and Europe for the first time. The results suggest a low genetic diversity within USUV, the existence of a distinct USUV subtype strain, and support the hypothesis that USUV was introduced to Europe from Africa. Further sequencing and analysis of USUV isolates from other African countries would contribute to a better understanding of its genetic diversity and geographic distribution. PMID:23816256

2013-01-01

341

Characterization of sour cherry isolates of plum pox virus from the Volga Basin in Russia reveals a new cherry strain of the virus.  

PubMed

Plum pox virus (PPV) is the causal agent of sharka, the most detrimental virus disease of stone fruit trees worldwide. PPV isolates have been assigned into seven distinct strains, of which PPV-C regroups the genetically distinct isolates detected in several European countries on cherry hosts. Here, three complete and several partial genomic sequences of PPV isolates from sour cherry trees in the Volga River basin of Russia have been determined. The comparison of complete genome sequences has shown that the nucleotide identity values with other PPV isolates reached only 77.5 to 83.5%. Phylogenetic analyses clearly assigned the RU-17sc, RU-18sc, and RU-30sc isolates from cherry to a distinct cluster, most closely related to PPV-C and, to a lesser extent, PPV-W. Based on their natural infection of sour cherry trees and genomic characterization, the PPV isolates reported here represent a new strain of PPV, for which the name PPV-CR (Cherry Russia) is proposed. The unique amino acids conserved among PPV-CR and PPV-C cherry-infecting isolates (75 in total) are mostly distributed within the central part of P1, NIa, and the N terminus of the coat protein (CP), making them potential candidates for genetic determinants of the ability to infect cherry species or of adaptation to these hosts. The variability observed within 14 PPV-CR isolates analyzed in this study (0 to 2.6% nucleotide divergence in partial CP sequences) and the identification of these isolates in different localities and cultivation conditions suggest the efficient establishment and competitiveness of the PPV-CR in the environment. A specific primer pair has been developed, allowing the specific reverse-transcription polymerase chain reaction detection of PPV-CR isolates. PMID:23581702

Glasa, Miroslav; Prikhodko, Yuri; Predaj?a, Lukáš; Nagyová, Alžbeta; Shneyder, Yuri; Zhivaeva, Tatiana; Subr, Zdeno; Cambra, Mariano; Candresse, Thierry

2013-09-01

342

The Isolation of Cricket Paralysis Virus from the Emperor Gum Moth, Antheraea eucalypti Scott, and its Infectivity towards a Range of Insect Species  

Microsoft Academic Search

Summary Three virus-like particles have been isolated from diseased larvae of Antheraea eucalypti. Serological tests established that one of them was indistinguishable from cricket paralysis virus (CrPV). CrPV isolated from crickets and from Antheraea were cross-infectious, and crickets could acquire lethal doses of the virus by feeding on infected Antheraea larvae. In addition to two species of Teleogryllus, three other

Carl Reinganum

1975-01-01

343

Complete Genome Sequence of a Novel Reassortant H3N6 Avian Influenza Virus Isolated from Domestic Green-Winged Teal  

PubMed Central

An avian influenza virus strain, A/domestic green-winged teal/Hunan/2036/2007(H3N6) (DGW-T2036), was isolated from healthy domestic green-winged teals (Anas crecca) in Hunan Province, South China. All eight gene segments of the isolate were sequenced. Genomic analysis demonstrated that this H3N6 virus is a novel reassortant avian influenza virus with a gene constellation originating from multiple ancestors. PMID:23682150

Xiong, Chaochao; Liu, Qian; Chen, Quanjiao; Yao, Yanfeng; Wang, Huadong

2013-01-01

344

Complete Genome Sequence of a Novel Reassortant H3N6 Avian Influenza Virus Isolated from Domestic Green-Winged Teal.  

PubMed

An avian influenza virus strain, A/domestic green-winged teal/Hunan/2036/2007(H3N6) (DGW-T2036), was isolated from healthy domestic green-winged teals (Anas crecca) in Hunan Province, South China. All eight gene segments of the isolate were sequenced. Genomic analysis demonstrated that this H3N6 virus is a novel reassortant avian influenza virus with a gene constellation originating from multiple ancestors. PMID:23682150

Xiong, Chaochao; Liu, Qian; Chen, Quanjiao; Yao, Yanfeng; Wang, Huadong; Chen, Jianjun

2013-01-01

345

Characterization of an influenza A virus isolated from pigs during an outbreak of respiratory disease in swine and people during a county fair in the United States  

Microsoft Academic Search

In August 2007, pigs and people became clinically affected by an influenza-like illness during attendance at an Ohio county fair. Influenza A virus was identified from pigs and people, and the virus isolates were characterized as swine H1N1 similar to swine viruses currently circulating in the U.S. pig population. The swine isolate, A\\/SW\\/OH\\/511445\\/2007 (OH07), was evaluated in an experimental challenge

Amy L. Vincent; Sabrina L. Swenson; Kelly M. Lager; Phillip C. Gauger; Christina Loiacono; Yan Zhang

2009-01-01

346

Characterization of a swine-like reassortant H1N2 influenza virus isolated from a wild duck in the United States  

Microsoft Academic Search

An H1N2 influenza virus (A\\/Duck\\/North Carolina\\/91347\\/01) (Dk\\/NC) was isolated from a wild duck in the United States in 2001. Genetic analyses showed that this duck virus has the same human\\/classical swine\\/avian reassortant genotype as the H1N2 viruses that have been isolated from pigs and turkeys in the US since 1999. Phylogenetic analyses of each gene segment further confirmed that the

Christopher W. Olsen; Alexander Karasin; Gene Erickson

2003-01-01

347

Amantadine Resistance among Porcine H1N1, H1N2, and H3N2 Influenza A Viruses Isolated in Germany between 1981 and 2001  

Microsoft Academic Search

This study was designed to gain insight into amantadine susceptibility of porcine influenza A viruses isolated in Germany between 1981 and 2001. The 12 studied H1N1, H1N2, and H3N2 porcine influenza virus strains were isolated in chicken eggs and passaged once in MDCK cells. Plaque reduction assays were applied to examine virus susceptibility to amantadine. Genotyping was used to confirm

Michaela Schmidtke; Roland Zell; Katja Bauer; Andi Krumbholz; Christina Schrader; Jochen Suess; Peter Wutzler

2006-01-01

348

Comparison of Newcastle disease viruses isolated from cormorants in Canada and the USA in 1975, 1990 and 1992.  

PubMed Central

Seventeen Newcastle disease virus (NDV) isolates obtained from cormorants, turkeys, a pelican, and a gull in Canada and the USA collected in 1975, 1990 and 1992 were analyzed for relatedness by monoclonal antibody profiling. In addition, nucleotide sequence analysis was performed in two areas of the fusion (F) gene for 5 of the isolates. No difference in the antigenicity of these 17 viruses, as determined by monoclonal antibody binding patterns, was seen. The amino acid sequences obtained via nucleotide sequencing at the cleavage site of the F protein showed that all the isolates tested had two pairs of basic amino acids immediately upstream of the cleavage site, and a phenylalanine residue at the N-terminus of the F1 protein, which is consistent with velogenic NDV. The deduced amino acid sequence obtained at the cleavage site of the F protein from 6 of the isolates was virtually identical regardless of the species, year of isolation, or location. However, the 1975 cormorant isolate showed marked differences from the 1990-1992 isolates in the nucleotide and deduced amino acid sequence of the F gene signal region. These data indicate that the 1990 and 1992 outbreaks were caused by the same epizootic virus and further suggest that the population of NDV in these wild birds may be very stable. The belief that the velogenic NDV circulating in cormorants in 1992 was transmitted into the free-ranging turkey flocks located near the cormorants in North Dakota is supported by the present study in which no distinction could be made between the viruses isolated from turkeys or wild birds. PMID:8825994

Heckert, R A; Collins, M S; Manvell, R J; Strong, I; Pearson, J E; Alexander, D J

1996-01-01

349

A unique segment of the hepatitis B virus group A genotype identified in isolates from South Africa  

Microsoft Academic Search

The preS2\\/S genes of hepatitis B virus isolated from 29 acutely or chronically infected individuals in the Gauteng province of South Africa were sequenced. Phylogenetic analysis of these sequences in com- parison with global isolates from the GenBank database showed that 24 sequences clustered with genotypic group A, three with genotypic group D and one each with genotypic groups B

Sheila M. Bowyer; Louise van Staden; Michael C. Kew; John G. M. Sim

350

Isolation and sequencing of the Epstein-Barr virus BNLF-1 gene (LMP1) from a Chinese nasopharyngeal carcinoma  

Microsoft Academic Search

The BamHI fragment containing the Epstein-Barr virus (EBV) LMP1 gene was cloned from a genomic library of the nude mouse-propagated Chinese naso- pharyngeal carcinoma CAO. The sequence of the LMP1 gene and its promoter and enhancer was determined. The nucleotide sequence of the CAO isolate differed from those of the B95-8 and Raji isolates in the promoter\\/enhancer region; the amino

Li-Fu Hu; Eugene R. Zabarovsky; Fu Chen; Shi-Long Cao; Ingemar Ernberg; George Klein; G. Winberg

1991-01-01

351

Comparative analysis of the genomes of two isolates of cowpea aphid-borne mosaic virus (CABMV) obtained from different hosts  

Microsoft Academic Search

The complete genomic sequences of two cowpea aphid-borne mosaic virus (CABMV) isolates from Brazil, MG-Avr from passion fruit (which also infects cowpea), and BR1 from peanut (which also infects cowpea, but not passion fruit), were determined. Their nucleotide sequences are 89% identical and display 85% identity to that of CABMV-Z. Both isolates have the typical potyvirus genome features. P3 and

Danielle R. Barros; Poliane Alfenas-Zerbini; José Evando A. Beserra; Tathiana F. S. Antunes; F. Murilo Zerbini

2011-01-01

352

Molecular characterization of highly pathogenic H5N1 avian influenza viruses isolated in Sweden in 2006  

PubMed Central

Background The analysis of the nonstructural (NS) gene of the highly pathogenic (HP) H5N1 avian influenza viruses (AIV) isolated in Sweden early 2006 indicated the co-circulation of two sub-lineages of these viruses at that time. In order to complete the information on their genetic features and relation to other HP H5N1 AIVs the seven additional genes of twelve Swedish isolates were amplified in full length, sequenced, and characterized. Results The presence of two sub-lineages of HP H5N1 AIVs in Sweden in 2006 was further confirmed by the phylogenetic analysis of approximately the 95% of the genome of twelve isolates that were selected on the base of differences in geographic location, timing and animal species of origin. Ten of the analyzed viruses belonged to sub-clade 2.2.2. and grouped together with German and Danish isolates, while two 2.2.1. sub-clade viruses formed a cluster with isolates of Egyptian, Italian, Slovenian, and Nigerian origin. The revealed amino acid differences between the two sub-groups of Swedish viruses affected the predicted antigenicity of the surface glycoproteins, haemagglutinin and neuraminidase, rather than the nucleoprotein, polymerase basic protein 2, and polymerase acidic protein, the main targets of the cellular immune responses. The distinctive characteristics between members of the two subgroups were identified and described. Conclusion The comprehensive genetic characterization of HP H5N1 AIVs isolated in Sweden during the spring of 2006 is reported. Our data support previous findings on the coincidental spread of multiple sub-lineage H5N1 HPAIVs via migrating aquatic birds to large distance from their origin. The detection of 2.2.1. sub-clade viruses in Sweden adds further data regarding their spread in the North of Europe in 2006. The close genetic relationship of Swedish isolates sub-clade 2.2.2. to the contemporary German and Danish isolates supports the proposition of the introduction and spread of a single variant of 2.2.2. sub-clade H5N1 avian influenza viruses in the Baltic region. The presented findings underline the importance of whole genome analysis. PMID:18837987

Kiss, István; Gyarmati, Péter; Zohari, Siamak; Ramsay, Karin Wilbe; Metreveli, Giorgi; Weiss, Elisabeth; Brytting, Maria; Stivers, Marielle; Lindström, Sofia; Lundkvist, Ake; Nemirov, Kirill; Thorén, Peter; Berg, Mikael; Czifra, György; Belák, Sándor

2008-01-01

353

Avian Influenza Virus Isolated in Wild Waterfowl in Argentina: Evidence of a potentially unique phylogenetic lineage in South America  

PubMed Central

Avian Influenza (AI) viruses have been sporadically isolated in South America. The most recent reports are from an outbreak in commercial poultry in Chile in 2002 and its putative ancestor from a wild bird in Bolivia in 2001. Extensive surveillance in wild birds was carried out in Argentina during 2006-2007. Using RRT-PCR, 12 AI positive detections were made from cloacal swabs. One of those positive samples yielded an AI virus isolated from a wild kelp gull (Larus dominicanus) captured in the South Atlantic coastline of Argentina. Further characterization by nucleotide sequencing reveals that it belongs to the H13N9 subtype. Phylogenetic analysis of the 8 viral genes suggests that the 6 internal genes are related to the isolates from Chile and Bolivia. The analysis also indicates that a cluster of phylogenetically related AI viruses from South America may have evolved independently, with minimal gene exchange, from influenza viruses in other latitudes. The data produced from our investigations are valuable contributions to the study of AI viruses in South America. PMID:18632129

Pereda, Ariel J.; Uhart, Marcela; Perez, Alberto A.; Zaccagnini, Maria E.; La Sala, Luciano; Decarre, Julieta; Goijman, Andrea; Solari, Laura; Suarez, Romina; Craig, Maria I.; Vagnozzi, Ariel; Rimondi, Agustina; Konig, Guido; Terrera, Maria V.; Kaloghlian, Analia; Song, Haichen; Sorrell, Erin M.; Perez, Daniel R.

2008-01-01

354

“Sakhalin” virus — a new arbovirus isolated from Ixodes (Ceratixodes) putus Pick.-Camb. 1878 collected on Tuleniy Island, Sea of Okhotsk  

Microsoft Academic Search

Summary Fifteen antigenically related virus strains isolated in 1969–1971 fromIxodes putus ticks at a seabird colony (Uria aalge) of the south-east coast of Sakhalin Island, Sea of Okhotsk, were studied. Sakhalin virus was shown to be a RNA-containing virus, sensitive to ether and sodium deoxycholate, pathogenic for infant mice after intracerebral inoculation. The virus did not agglutinate goose erythrocytes. No

D. K. Lvov; A. A. Timofeeva; V. L. Gromashevski; V. I. Chervonsky; A. I. Gromov; Yu. M. Tsyrkin; A. G. Pogrebenko; I. N. Kostyrko

1972-01-01

355

Mosquito surveillance for West Nile virus in Connecticut, 2000: isolation from Culex pipiens, Cx. restuans, Cx. salinarius, and Culiseta melanura.  

PubMed Central

Fourteen isolations of West Nile (WN) virus were obtained from four mosquito species (Culex pipiens [5], Cx. restuans [4], Cx. salinarius [2], and Culiseta melanura [3]) in statewide surveillance conducted from June through October 2000. Most isolates were obtained from mosquitoes collected in densely populated residential locales in Fairfield and New Haven counties, where the highest rates of dead crow sightings were reported and where WN virus was detected in 1999. Minimum field infection rates per 1,000 mosquitoes ranged from 0.5 to 1.8 (county based) and from 1.3 to 76.9 (site specific). Cx. restuans appears to be important in initiating WN virus transmission among birds in early summer; Cx. pipiens appears to play a greater role in amplifying virus later in the season. Cs. melanura could be important in the circulation of WN virus among birds in sylvan environments; Cx. salinarius is a suspected vector of WN virus to humans and horses. PMID:11585530

Andreadis, T. G.; Anderson, J. F.; Vossbrinck, C. R.

2001-01-01

356

Wild-type temperature-sensitive and -resistant visna viruses: isolation and biological comparison.  

PubMed

The plaques formed by wild-type (WT) populations of strain K485 visna (V-K485) virus, strain K796 visna (V-K796) virus, and of progressive pneumonia virus in sheep choroid plexus (SCP) cell cultures are heterogeneous in size. Plaque purification procedures showed that this heterogeneity was due to the presence of two biologically different viruses, large plaque-forming (Lpf) virus and small plaque-forming (Spf) virus. The V-K796 Lpf virus, the V-K796 Spf virus, and the V-K796 WT virus are antigenically similar to each other. Although the V-K796 Spf virus is less cytopathic than the V-K796 Lpf virus, both viruses have a 14- to 16-hour latent period in SCP cells and have similar rates of synthesis for the initial 24 to 30 hours after infection. The V-K796 Spf virus is considered a temperature-sensitive visna virus since, unlike the V-K796 Lpf virus, it does not produce plaques at 41 degrees C and it is more thermosensitive than the V-K796 Lpf variant. Population analyses of two strains of wild-type visna virus and one strain of progressive pneumonia virus demonstrated that these populations contain variants with the phenotypes of V-K796 Spf virus and V-K796 Lpf virus, that the Spf virus was not selected against in sheep, and that cultivation of the wild type visna viruses at 37 degrees C selected against the temperature-sensitive variants. The existence of the temperature-sensitive Spf viruses in these wild-type virus populations suggests that the Spf variants maintain the persistent visna virus infection in sheep and delay the development of the host's cell-mediated immune response to the viral proteins. PMID:6261720

Trowbridge, R S; Lehmann, J; Torchio, C; Brophy, P

1981-01-01

357

Isolation and analysis of a very virulent Marek's disease virus strain in China  

PubMed Central

Background A severe MD was broken out at a farm in Shandong, China, despite FC126 vaccination of the chickens at 1-day-old. The mortality of the flocks reached up to 38.3%. The infected chickens were found to have MD pathological changes, including enlargement of spleens, livers and kidneys, and tumors occured on organs later. Samples were collected from the chickens for diagnosis. Methods The collected samples were inoculated into primary duck embryo fibroblast (DEF) cells, and the MDV strain named SD2012-1 was isolated. In order to identify the isolate, amplification by PCR and sequencing of oncogenic Meq and vIL-8 gene were processed, the obtained sequences were compared with the sequences of reference strains, and SD2012-1 was used to challenge immunized SPF chickens. Results A very virulent MDV isolate strain, SD2012-1, was isolated from a chicken flock in Shandong Province, China, the isolate had the characteristics of very virulent MDV-1, nucleotide and deduced amino acid sequence comparisons of Meq and vIL-8 gene of SD2012-1 with those of reference strains showed SD2012-1 had high homology with MDV strains isolated from China, SD2012-1 could break through the protection provided by HVT vaccine and HVT?+?SB-1 vaccine immunization and caused the mortality of SPF chickens over 60%. The immune failure occured at the farm could be due to the improper selection of vaccines. SD2012-1 produced death later and the gross postmortem lesions of chickens died early and later were different. Conclusions MDV strain SD2012-1 isolated from Shandong Province, China was found to have the characteristics of very virulent MDV-1, which could break through the protection provided by HVT vaccine and HVT?+?SB-1 vaccine, the virus seemed to have a long latent period, and cause different gross postmortem lesions of chickens between chickens died early and later. A better immunization way should be chosen to prevent infection of this MDV strain in field. PMID:23687964

2013-01-01

358

Isolation and Characterization of the Herpes Simplex Virus 1 Terminase Complex  

PubMed Central

During herpes simplex virus 1 (HSV-1) infection, empty procapsids are assembled and subsequently filled with the viral genome by means of a protein complex called the terminase, which is comprised of the HSV-1 UL15, UL28, and UL33 proteins. Biochemical studies of the terminase proteins have been hampered by the inability to purify the intact terminase complex. In this study, terminase complexes were isolated by tandem-affinity purification (TAP) using recombinant viruses expressing either a full-length NTAP-UL28 fusion protein (vFH476) or a C-terminally truncated NTAP-UL28 fusion protein (vFH499). TAP of the UL28 protein from vFH476-infected cells, followed by silver staining, Western blotting, and mass spectrometry, identified the UL15, UL28, and UL33 subunits, while TAP of vFH499-infected cells confirmed previous findings that the C terminus of UL28 is required for UL28 interaction with UL33 and UL15. Analysis of the oligomeric state of the purified complexes by sucrose density gradient ultracentrifugation revealed that the three proteins formed a complex with a molecular mass that is consistent with the formation of a UL15-UL28-UL33 heterotrimer. In order to assess the importance of conserved regions of the UL15 and UL28 proteins, recombinant NTAP-UL28 viruses with mutations of the putative UL28 metal-binding domain or within the UL15 nuclease domain were generated. TAP of UL28 complexes from cells infected with each domain mutant demonstrated that the conserved cysteine residues of the putative UL28 metal-binding domain and conserved amino acids within the UL15 nuclease domain are required for the cleavage and packaging functions of the viral terminase, but not for terminase complex assembly. PMID:24155374

Heming, Jason D.; Huffman, Jamie B.; Jones, Lisa M.

2014-01-01

359

Broome virus, a new fusogenic Orthoreovirus species isolated from an Australian fruit bat  

SciTech Connect

This report describes the discovery and characterization of a new fusogenic orthoreovirus, Broome virus (BroV), isolated from a little red flying-fox (Pteropus scapulatus). The BroV genome consists of 10 dsRNA segments, each having a 3' terminal pentanucleotide sequence conserved amongst all members of the genus Orthoreovirus, and a unique 5' terminal pentanucleotide sequence. The smallest genome segment is bicistronic and encodes two small nonstructural proteins, one of which is a novel fusion associated small transmembrane (FAST) protein responsible for syncytium formation, but no cell attachment protein. The low amino acid sequence identity between BroV proteins and those of other orthoreoviruses (13-50%), combined with phylogenetic analyses of structural and nonstructural proteins provide evidence to support the classification of BroV in a new sixth species group within the genus Orthoreovirus.

Thalmann, Claudia M. [CSIRO Livestock Industries, Australian Animal Health Laboratory, Geelong, Victoria (Australia); Australian Biosecurity Cooperative Research Centre for Emerging Infectious Disease, Brisbane, Queensland (Australia); School of Chemistry and Molecular Biosciences, University of Queensland, St Lucia, Queensland (Australia); Cummins, David Michael [CSIRO Livestock Industries, Australian Animal Health Laboratory, Geelong, Victoria (Australia); Yu Meng [CSIRO Livestock Industries, Australian Animal Health Laboratory, Geelong, Victoria (Australia); Australian Biosecurity Cooperative Research Centre for Emerging Infectious Disease, Brisbane, Queensland (Australia); Lunt, Ross; Pritchard, Lindsay Ian; Hansson, Eric; Crameri, Sandra; Hyatt, Alex [CSIRO Livestock Industries, Australian Animal Health Laboratory, Geelong, Victoria (Australia); Wang Linfa, E-mail: Linfa.Wang@csiro.a [CSIRO Livestock Industries, Australian Animal Health Laboratory, Geelong, Victoria (Australia); Australian Biosecurity Cooperative Research Centre for Emerging Infectious Disease, Brisbane, Queensland (Australia)

2010-06-20

360

Isolation and Genetic Analysis of Bovine Viral Diarrhea Virus from Infected Cattle in Indiana  

PubMed Central

Species and biotype distribution was determined in 44 bovine viral diarrhea virus- (BVDV-) positive samples submitted to the Animal Disease Diagnostic Laboratory (ADDL) in Indiana during 2006–2008. BVDV RNA was detected in the 5?-untranslated region and Npro region using reverse transcriptase PCR followed by sequencing analysis of the PCR product. Additionally, cases were classified into one of six categories according to history and/or lesions: acute symptomatic, hemorrhagic, respiratory distress, reproductive, persistent infection (PI), and mucosal disease (MD). Of 44 BVDV-positive samples, 33 were noncytopathic (ncp), 10 were cytopathic (cp), and one presented both ncp and cp biotypes. Sequencing analysis demonstrated that all samples belonged to BVDV-1a, BVDV-1b, or BVDV-2. The most common isolate was ncp BVDV-1b, (44%) followed by ncp BVDV-2a (24%). Among the six categories, respiratory clinical signs were the most common (36%) followed by PI (25%) and MD (16%). PMID:21647344

Pogranichniy, Roman M.; Schnur, Megan E.; Raizman, Eran A.; Murphy, Duane A.; Negron, Maria; Thacker, H. Leon

2011-01-01

361

Niakha virus: A novel member of the family Rhabdoviridae isolated from phlebotomine sandflies in Senegal  

PubMed Central

Members of the family Rhabdoviridae have been assigned to eight genera but many remain unassigned. Rhabdoviruses have a remarkably diverse host range that includes terrestrial and marine animals, invertebrates and plants. Transmission of some rhabdoviruses often requires an arthropod vector, such as mosquitoes, midges, sandflies, ticks, aphids and leafhoppers, in which they replicate. Herein we characterize Niakha virus (NIAV), a previously uncharacterized rhabdovirus isolated from phebotomine sandflies in Senegal. Analysis of the 11,124 nt genome sequence indicates that it encodes the five common rhabdovirus proteins with alternative ORFs in the M, G and L genes. Phylogenetic analysis of the L protein indicate that NIAV’s closest relative is Oak Vale rhabdovirus, although in this analysis NIAV is still so phylogenetically distinct that it might be classified as distinct from the eight currently recognized Rhabdoviridae genera. This observation highlights the vast, and yet not fully recognized diversity, of this family. PMID:23773405

Vasilakis, Nikos; Widen, Steven; Mayer, Sandra V.; Seymour, Robert; Wood, Thomas G.; Popov, Vsevolov; Guzman, Hilda; da Rosa, Amelia P.A. Travassos; Ghedin, Elodie; Holmes, Edward C.; Walker, Peter J.; Tesh, Robert B.

2013-01-01

362

Isolation of Newcastle disease virus and Salmonella typhimurium from the brain of double-crested cormorants (Phalacrocorax auritus).  

PubMed

Avian paramyxovirus type 1 (Newcastle disease virus) and Salmonella typhimurium were isolated from the brain and lung tissues of double-crested cormorants (Phalacrocorax auritus) from Lac Canard, Alberta, Canada. More than 100 birds died during this outbreak in 1999. Affected birds presented signs of central nervous system disease characterized by unilateral wing and leg paralysis. Other geographic locations in the provinces of Alberta and Saskatchewan have reported cases of cormorants suffering from diseases with signs compatible with Newcastle disease. The virus isolated in the 1999 outbreak was characterized as mesogenic. These findings suggest that other pathogens, like S. typhimurium, may influence the clinical presentation of disease caused by mesogenic strains of Newcastle disease virus in cormorants. PMID:11332490

Clavijo, A; Robinson, Y; López, J

2001-01-01

363

Isolation of Erysipelothrix rhusiopathiae from a red-tailed hawk (Buteo jamaicensis) with a concurrent pox virus infection.  

PubMed

Erysipelothrix rhusiopathiae was isolated from the spleen, liver, lung, heart, kidney, and skin of a red-tailed hawk (Buteo jamaicensis) which had a concurrent avian pox virus infection. The hawk had been housed on a farm with domestic turkeys, providing a possible source of the E. rhusiopathiae. PMID:2824865

Pace, L W; Chengappa, M M; Greer, S; Alderson, C

1987-10-01

364

Pathogenesis of Primary Respiratory Disease Induced by Isolates from a New Genetic Cluster of Bovine Viral Diarrhea Virus Type I  

Microsoft Academic Search

The pathogenesis of infection induced by cytopathogenic isolates from the newly identified genetic cluster Id of bovine viral diarrhea virus (BVDV) type I was studied in two experimental infections of previously sero- negative, immunocompetent calves. Experiment 1 focused on the evaluation of clinical patterns, viremia, and serological responses. All infected calves in this experiment developed respiratory symptoms and seroconverted to

C. Baule; G. Kulcsar; K. Belak; M. Albert; C. Mittelholzer; T. Soos; L. Kucsera; S. Belak

2001-01-01

365

Regional Clustering of Shared Neutralization Determinants on Primary Isolates of Clade C Human Immunodeficiency Virus Type 1 from South Africa  

Microsoft Academic Search

Clade C is one of the most prevalent genetic subtypes of human immunodeficiency virus type 1 (HIV-1) in the world today and one of the least studied with respect to neutralizing antibodies. Most information on HIV-1 serology as it relates to neutralization is derived from clade B. Clade C primary isolates of HIV-1 from South Africa and Malawi were shown

Renata Bures; Lynn Morris; Carolyn Williamson; Gita Ramjee; Mark Deers; Susan A. Fiscus; Salim Abdool-Karim; David C. Montefiori

2002-01-01

366

Complete genome sequence of an Indian field isolate of classical Swine Fever virus belonging to subgenotype 1.1.  

PubMed

We report the complete genome sequence of an Indian field isolate of classical swine fever virus (CSFV) belonging to predominant subgenotype 1.1 prevalent in India. This report will help in understanding the molecular diversity of CSFV strains circulating worldwide and to select and develop a suitable vaccine candidate for classical swine fever (CSF) control in India. PMID:25278522

Kamboj, Aman; Patel, Chhabi L; Chaturvedi, V K; Saini, Mohini; Gupta, Praveen K

2014-01-01

367

Isolation and characterization of porcine epidemic diarrhea viruses associated with the 2013 disease outbreak among swine in the United States.  

PubMed

Porcine epidemic diarrhea virus (PEDV) was detected in May 2013 for the first time in U.S. swine and has since caused significant economic loss. Obtaining a U.S. PEDV isolate that can grow efficiently in cell culture is critical for investigating pathogenesis and developing diagnostic assays and for vaccine development. An additional objective was to determine which gene(s) of PEDV is most suitable for studying the genetic relatedness of the virus. Here we describe two PEDV isolates (ISU13-19338E and ISU13-22038) successfully obtained from the small intestines of piglets from sow farms in Indiana and Iowa, respectively. The two isolates have been serially propagated in cell culture for over 30 passages and were characterized for the first 10 passages. Virus production in cell culture was confirmed by PEDV-specific real-time reverse-transcription PCR (RT-PCR), immunofluorescence assays, and electron microscopy. The infectious titers of the viruses during the first 10 passages ranged from 6 × 10(2) to 2 × 10(5) 50% tissue culture infective doses (TCID50)/ml. In addition, the full-length genome sequences of six viruses (ISU13-19338E homogenate, P3, and P9; ISU13-22038 homogenate, P3, and P9) were determined. Genetically, the two PEDV isolates were relatively stable during the first 10 passages in cell culture. Sequences were also compared to those of 4 additional U.S. PEDV strains and 23 non-U.S. strains. All U.S. PEDV strains were genetically closely related to each other (?99.7% nucleotide identity) and were most genetically similar to Chinese strains reported in 2011 to 2012. Phylogenetic analyses using different genes of PEDV suggested that the full-length spike gene or the S1 portion is appropriate for sequencing to study the genetic relatedness of these viruses. PMID:24197882

Chen, Qi; Li, Ganwu; Stasko, Judith; Thomas, Joseph T; Stensland, Wendy R; Pillatzki, Angela E; Gauger, Phillip C; Schwartz, Kent J; Madson, Darin; Yoon, Kyoung-Jin; Stevenson, Gregory W; Burrough, Eric R; Harmon, Karen M; Main, Rodger G; Zhang, Jianqiang

2014-01-01

368

Measurement of the Infection and Dissemination of Bluetongue Virus in Culicoides Biting Midges Using a Semi-Quantitative RT-PCR Assay and Isolation of Infectious Virus  

PubMed Central

Background Culicoides biting midges (Diptera: Ceratopogonidae) are the biological vectors of globally significant arboviruses of livestock including bluetongue virus (BTV), African horse sickness virus (AHSV) and the recently emerging Schmallenberg virus (SBV). From 2006–2009 outbreaks of BTV in northern Europe inflicted major disruption and economic losses to farmers and several attempts were made to implicate Palaearctic Culicoides species as vectors. Results from these studies were difficult to interpret as they used semi-quantitative RT-PCR (sqPCR) assays as the major diagnostic tool, a technique that had not been validated for use in this role. In this study we validate the use of these assays by carrying out time-series detection of BTV RNA in two colony species of Culicoides and compare the results with the more traditional isolation of infectious BTV on cell culture. Methodology/Principal Findings A BTV serotype 1 strain mixed with horse blood was fed to several hundred individuals of Culicoides sonorensis (Wirth & Jones) and C. nubeculosus (Mg.) using a membrane-based assay and replete individuals were then incubated at 25°C. At daily intervals 25 Culicoides of each species were removed from incubation, homogenised and BTV quantified in each individual using sqPCR (Cq values) and virus isolation on a KC-C. sonorensis embryonic cell line, followed by antigen enzyme-linked immunosorbent assay (ELISA). In addition, comparisons were also drawn between the results obtained with whole C. sonorensis and with individually dissected individuals to determine the level of BTV dissemination. Conclusions/Significance Cq values generated from time-series infection experiments in both C. sonorensis and C. nubeculosus confirmed previous studies that relied upon the isolation and detection of infectious BTV. Implications on the testing of field-collected Culicoides as potential virus vectors by PCR assays and the use of such assays as front-line tools for use in diagnostic laboratories in this role are discussed. PMID:23940643

Veronesi, Eva; Antony, Frank; Gubbins, Simon; Golding, Nick; Blackwell, Alison; Mertens, Peter PC.; Brownlie, Joe; Darpel, Karin E.; Mellor, Philip S.; Carpenter, Simon

2013-01-01

369

Receptor Specificity and Transmission of H2N2 Subtype Viruses Isolated from the Pandemic of 1957  

PubMed Central

Influenza viruses of the H2N2 subtype have not circulated among humans in over 40 years. The occasional isolation of avian H2 strains from swine and avian species coupled with waning population immunity to H2 hemagglutinin (HA) warrants investigation of this subtype due to its pandemic potential. In this study we examined the transmissibility of representative human H2N2 viruses, A/Albany/6/58 (Alb/58) and A/El Salvador/2/57 (ElSalv/57), isolated during the 1957/58 pandemic, in the ferret model. The receptor binding properties of these H2N2 viruses was analyzed using dose-dependent direct glycan array-binding assays. Alb/58 virus, which contains the 226L/228S amino acid combination in the HA and displayed dual binding to both alpha 2,6 and alpha 2,3 glycan receptors, transmitted efficiently to naďve ferrets by respiratory droplets. Inefficient transmission was observed with ElSalv/57 virus, which contains the 226Q/228G amino acid combination and preferentially binds alpha 2,3 over alpha 2,6 glycan receptors. However, a unique transmission event with the ElSalv/57 virus occurred which produced a 226L/228G H2N2 natural variant virus that displayed an increase in binding specificity to alpha 2,6 glycan receptors and enhanced respiratory droplet transmissibility. Our studies provide a correlation between binding affinity to glycan receptors with terminal alpha 2,6-linked sialic acid and the efficiency of respiratory droplet transmission for pandemic H2N2 influenza viruses. PMID:20574518

Pappas, Claudia; Viswanathan, Karthik; Chandrasekaran, Aarthi; Raman, Rahul; Katz, Jacqueline M.; Sasisekharan, Ram; Tumpey, Terrence M.

2010-01-01

370

Transfer of the 3' non-translated region of grapevine chrome mosaic virus RNA-1 by recombination to tomato black ring virus RNA-2 in pseudorecombinant isolates.  

PubMed

In grapevine chrome mosaic and tomato black ring viruses (GCMV and TBRV), as in many other nepoviruses, the 3' non-translated regions (3'NTR) are identical between the two genomic RNAs. We have investigated the structure of the 3'NTR of two recombinant isolates which contain GCMV RNA-1 and TBRV RNA-2. In these isolates, the 3'NTR of RNA-1 was transferred to RNA-2, thus restoring the 3' identity. The transfer occurred within three passages, and probably contributes to the spread of randomly appearing mutations from one genomic RNA to the other. The site of recombination is near the 3' end of the open reading frame. PMID:7730815

Le Gall, O; Candresse, T; Dunez, J

1995-05-01

371

The complete nucleotide sequence of Alternanthera mosaic virus infecting Portulaca grandiflora represents a new strain distinct from phlox isolates.  

PubMed

A southeastern European isolate of Alternanthera mosaic virus (AltMV-MU) of the genus Potexvirus (family Flexiviridae) was purified from the ornamental plant Portulaca grandiflora. The complete nucleotide sequence (6606 nucleotides) of AltMV-MU genomic RNA was defined. The AltMV-MU genome is different from those of all isolates described earlier and is most closely related to genomes of partly sequenced portulaca isolates AltMV-Po (America) and AltMV-It (Italy). Phylogenetic analysis supports the view that AltMV-MU belongs to a new "portulaca" genotype distinguishable from the "phlox" genotype. PMID:21127957

Ivanov, Peter A; Mukhamedzhanova, Anna A; Smirnov, Alexander A; Rodionova, Nina P; Karpova, Olga V; Atabekov, Joseph G

2011-04-01

372

Performance of virus isolation and Directigen® Flu A to detect influenza A virus in experimental human infection  

Microsoft Academic Search

Background: few data exist to assess the sensitivity of different specimen types for viral detection during the course of influenza virus infection. Objectives: this study assessed the relationships between quantitative influenza A virus replication and antigen detectability by the enzyme immunosorbent assay (EIA) Directigen® Flu A in different type of samples during experimental human infection. Study design: fourteen volunteers were

Laurent Kaiser; Marcus S Briones; Frederick G Hayden

1999-01-01

373

Characterization and distribution of two new foamy viruses isolated from chimpanzees  

Microsoft Academic Search

Summary Pan 1 and Pan 2 viruses are persistent viruses recoverable from many organs, including brain, of most chimpanzees; both may be present in the same organ of a given animal. These viruses induce a vacuolated, foamy syncytia without inclusion bodies both in chimpanzee explant cultures and in primary human embryo kidney (HEK) cells. Both viruses are not inhibited by

John J. Hooks; Clarence J. Gibbs; E. C. Cutchins; N. G. Rogers; P. Lampert; D. C. Gajdusek

1972-01-01

374

Full-length sequence analysis of a distinct isolate of Bidens mottle virus infecting sunflower in Taiwan.  

PubMed

The full-length genome of a potyvirus, previously known as sunflower chlorotic spot virus isolate SF-1 (SCSV-SF-1) which causes novel symptoms on sunflowers (Helianthus annuus), was sequenced and analyzed. The genome of SCSV-SF-1 is 9,741 nucleotides long, encoding a polyprotein of 3,071 amino acids containing the consensus motifs of potyviruses. Sequence comparison revealed that the 3'-terminus of SCSV-SF-1 shared over 96% similarities with isolates of Bidens mottle virus (BiMoV). However, SCSV-SF-1 has a very narrow host range, excluding the diagnostic host species for BiMoV, Bidens pilosa and Zinnia elegans. Therefore, SCSV-SF-1 is a distinct isolate of BiMoV. This is the first report of the full-length nucleotide sequence of BiMoV infecting sunflower in Taiwan. PMID:19308314

Liao, J Y; Hu, Chung-Chi; Chen, C C; Chang, C H; Deng, T C

2009-01-01

375

Complete genome sequence analysis of Japanese encephalitis virus isolated from a horse in India.  

PubMed

The complete genome of the Japanese encephalitis virus (JEV) strain JEV/eq/India/H225/2009(H225), isolated from an infected horse in India, was sequenced and compared to previously published JEV genomes. H225 genome was 10,977-nucleotides long, comprising a single ORF of 10,299-nucleotides, a 5'-UTR of 95 nucleotides and a 3'-UTR of 582 nucleotides. The H225 genome showed high levels of sequence identity with 47 fully sequenced JEV genomes, ranging from 99.3 % to 75.5 % for nucleotides and 99.2 % to 91.5 % for amino acid sequences. Phylogenetic analysis of the full-length sequence indicated that the H225 strain belongs to genotype III and is closely related to the Indian JEV strain Vellore P20778. A comparison of amino acids associated with neurovirulence in the E proteins and non-structural proteins of known virulent and attenuated JEV strains suggested H225 to be a highly virulent strain. This is the first report of whole-genome sequencing of a genotype III JEV genome isolated from equines. PMID:23001697

Singha, Harisankar; Gulati, Baldev R; Kumar, Prabhat; Singh, Birendra K; Virmani, Nitin; Singh, Raj K

2013-01-01

376

Expression, purification and characterization of the Lily symptomless virus coat protein from Lanzhou Isolate  

PubMed Central

Background Lily symptomless virus (LSV) is widespread in many countries where lily are grown or planted, and causes severe economic losses in terms of quantity and quality of flower and bulb production. To study the structure-function relationship of coat protein (CP) of LSV, to investigate antigenic relationships between coat protein subunits or intact virons, and to prepare specific antibodies against LSV, substantial amounts of CP protein are needed. Results Thus, full-length cDNA of LSV coat protein was synthesized and amplified by RT-PCR from RNA isolated from LSV Lanzhou isolate. The extended 33.6 kDa CP was cloned and expressed prokaryoticly and then purified by Ni-ion affinity chromatography. Its identity and antigenicity of recombinant CP were identified on Western-blotting by using the prepared anti-LSV antibodies. Conclusions The results indicate that fusion CP maintains its native antigenicity and specificity, providing a good source of antigen in preparation of LSV related antibodies. Detailed structural analysis of a pure recombinant CP should allow a better understanding of its role in cell attachment and LSV tropism. This investigation to LSV should provide some specific antibodies and aid to development a detection system for LSV diagnostics and epidemiologic surveys. PMID:20144245

2010-01-01

377

Characterization of Synthetic Chikungunya Viruses Based on the Consensus Sequence of Recent E1-226V Isolates  

PubMed Central

Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that re-emerged in 2004 and has caused massive outbreaks in recent years. The lack of a licensed vaccine or treatment options emphasize the need to obtain more insight into the viral life cycle and CHIKV-host interactions. Infectious cDNA clones are important tools for such studies, and for mechanism of action studies on antiviral compounds. Existing CHIKV cDNA clones are based on a single genome from an individual clinical isolate, which is expected to have evolved specific characteristics in response to the host environment, and possibly also during subsequent cell culture passaging. To obtain a virus expected to have the general characteristics of the recent E1-226V CHIKV isolates, we have constructed a new CHIKV full-length cDNA clone, CHIKV LS3, based on the consensus sequence of their aligned genomes. Here we report the characterization of this synthetic virus and a green fluorescent protein-expressing variant (CHIKV LS3-GFP). Their characteristics were compared to those of natural strain ITA07-RA1, which was isolated during the 2007 outbreak in Italy. In cell culture the synthetic viruses displayed phenotypes comparable to the natural isolate, and in a mouse model they caused lethal infections that were indistinguishable from infections with a natural strain. Compared to ITA07-RA1 and clinical isolate NL10/152, the synthetic viruses displayed similar sensitivities to several antiviral compounds. 3-deaza-adenosine was identified as a new inhibitor of CHIKV replication. Cyclosporin A had no effect on CHIKV replication, suggesting that cyclophilins -opposite to what was found for other +RNA viruses- do not play an essential role in CHIKV replication. The characterization of the consensus sequence-based synthetic viruses and their comparison to natural isolates demonstrated that CHIKV LS3 and LS3-GFP are suitable and representative tools to study CHIKV-host interactions, screen for antiviral compounds and unravel their mode of action. PMID:23936484

Scholte, Florine E. M.; Tas, Ali; Martina, Byron E. E.; Cordioli, Paolo; Narayanan, Krishna; Makino, Shinji; Snijder, Eric J.; van Hemert, Martijn J.

2013-01-01

378

Characterization of synthetic Chikungunya viruses based on the consensus sequence of recent E1-226V isolates.  

PubMed

Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that re-emerged in 2004 and has caused massive outbreaks in recent years. The lack of a licensed vaccine or treatment options emphasize the need to obtain more insight into the viral life cycle and CHIKV-host interactions. Infectious cDNA clones are important tools for such studies, and for mechanism of action studies on antiviral compounds. Existing CHIKV cDNA clones are based on a single genome from an individual clinical isolate, which is expected to have evolved specific characteristics in response to the host environment, and possibly also during subsequent cell culture passaging. To obtain a virus expected to have the general characteristics of the recent E1-226V CHIKV isolates, we have constructed a new CHIKV full-length cDNA clone, CHIKV LS3, based on the consensus sequence of their aligned genomes. Here we report the characterization of this synthetic virus and a green fluorescent protein-expressing variant (CHIKV LS3-GFP). Their characteristics were compared to those of natural strain ITA07-RA1, which was isolated during the 2007 outbreak in Italy. In cell culture the synthetic viruses displayed phenotypes comparable to the natural isolate, and in a mouse model they caused lethal infections that were indistinguishable from infections with a natural strain. Compared to ITA07-RA1 and clinical isolate NL10/152, the synthetic viruses displayed similar sensitivities to several antiviral compounds. 3-deaza-adenosine was identified as a new inhibitor of CHIKV replication. Cyclosporin A had no effect on CHIKV replication, suggesting that cyclophilins -opposite to what was found for other +RNA viruses- do not play an essential role in CHIKV replication. The characterization of the consensus sequence-based synthetic viruses and their comparison to natural isolates demonstrated that CHIKV LS3 and LS3-GFP are suitable and representative tools to study CHIKV-host interactions, screen for antiviral compounds and unravel their mode of action. PMID:23936484

Scholte, Florine E M; Tas, Ali; Martina, Byron E E; Cordioli, Paolo; Narayanan, Krishna; Makino, Shinji; Snijder, Eric J; van Hemert, Martijn J

2013-01-01

379

Molecular and Biological Characterization of Lettuce mosaic virus (LMV) Isolates Reveals a Distinct and Widespread Type of Resistance-Breaking Isolate: LMV-Most.  

PubMed

ABSTRACT Lettuce mosaic virus (LMV) causes an economically important seedborne and aphid-transmitted disease of lettuce and ornamental crops worldwide. The genetic diversity among 73 LMV isolates was examined based on a 216-nucleotide sequence at the variable region encoding the NIb-coat protein junction. Three clusters of LMV isolates were distinguished: LMV-Yar, LMV-Greek, and LMV-RoW. In the latter cluster, two subgroups of isolates, LMV-Common and LMV-Most, accounted for a large proportion of the LMV isolates analyzed. These two subgroups included the seedborne isolates, consistent with this property contributing a selective advantage and resulting in widespread distribution. In addition to being seedborne, LMV-Most isolates overcome the two resistance genes commonly used in lettuce, mo1(1) and mo1(2), and thus represent a potential threat to lettuce cultivation. The complete sequence of an LMV-Most isolate (LMV-AF199) was determined, allowing a better definition of the genetic relationships among LMV-Most, LMV-Common, and an additional isolate of the LMV-RoW cluster. PMID:18943032

Krause-Sakate, Renate; Le Gall, Olivier; Fakhfakh, Hatem; Peypelut, Martine; Marrakchi, Mohammed; Varveri, Christina; Pavan, Marcelo A; Souche, Sylvie; Lot, Hervé; Zerbini, F Murilo; Candresse, Thierry

2002-05-01

380

Molecular characterization of the Great Lakes viral hemorrhagic septicemia virus (VHSV) isolate from USA  

PubMed Central

Background Viral hemorrhagic septicemia virus (VHSV) is a highly contagious viral disease of fresh and saltwater fish worldwide. VHSV caused several large scale fish kills in the Great Lakes area and has been found in 28 different host species. The emergence of VHS in the Great Lakes began with the isolation of VHSV from a diseased muskellunge (Esox masquinongy) caught from Lake St. Clair in 2003. VHSV is a member of the genus Novirhabdovirus, within the family Rhabdoviridae. It has a linear single-stranded, negative-sense RNA genome of approximately 11 kbp, with six genes. VHSV replicates in the cytoplasm and produces six monocistronic mRNAs. The gene order of VHSV is 3'-N-P-M-G-NV-L-5'. This study describes molecular characterization of the Great Lakes VHSV strain (MI03GL), and its phylogenetic relationships with selected European and North American isolates. Results The complete genomic sequences of VHSV-MI03GL strain was determined from cloned cDNA of six overlapping fragments, obtained by RT-PCR amplification of genomic RNA. The complete genome sequence of MI03GL comprises 11,184 nucleotides (GenBank GQ385941) with the gene order of 3'-N-P-M-G-NV-L-5'. These genes are separated by conserved gene junctions, with di-nucleotide gene spacers. The first 4 nucleotides at the termini of the VHSV genome are complementary and identical to other novirhadoviruses genomic termini. Sequence homology and phylogenetic analysis show that the Great Lakes virus is closely related to the Japanese strains JF00Ehi1 (96%) and KRRV9822 (95%). Among other novirhabdoviruses, VHSV shares highest sequence homology (62%) with snakehead rhabdovirus. Conclusion Phylogenetic tree obtained by comparing 48 glycoprotein gene sequences of different VHSV strains demonstrate that the Great Lakes VHSV is closely related to the North American and Japanese genotype IVa, but forms a distinct genotype IVb, which is clearly different from the three European genotypes. Molecular characterization of the Great Lakes isolate will be helpful in studying the pathogenesis of VHSV using a reverse genetics approach and developing efficient control strategies. PMID:19852863

Ammayappan, Arun; Vakharia, Vikram N

2009-01-01

381

Phylogenetic Analysis of H6 Influenza Viruses Isolated from Rosy-Billed Pochards (Netta peposaca) in Argentina Reveals the Presence of Different HA Gene Clusters ?  

PubMed Central

Until recently, influenza A viruses from wild waterfowl in South America were rarely isolated and/or characterized. To explore the ecology of influenza A viruses in this region, a long-term surveillance program was established in 2006 for resident and migratory water birds in Argentina. We report the characterization of 5 avian influenza viruses of the H6 hemagglutinin (HA) subtype isolated from rosy-billed pochards (Netta peposaca). Three of these viruses were paired to an N2 NA subtype, while the other two were of the N8 subtype. Genetic and phylogenetic analyses of the internal gene segments revealed a close relationship with influenza viruses from South America, forming a unique clade and supporting the notion of independent evolution from influenza A viruses in other latitudes. The presence of NS alleles A and B was also identified. The HA and NA genes formed unique clades separate from North American and Eurasian viruses, with the exception of the HA gene of one isolate, which was more closely related to the North American lineage, suggesting possible interactions between viruses of North American and South American lineages. Animal studies suggested that these Argentine H6 viruses could replicate and transmit inefficiently in chickens, indicating limited adaptation to poultry. Our results highlight the importance of continued influenza virus surveillance in wild birds of South America, especially considering the unique evolution of these viruses. PMID:21976652

Rimondi, Agustina; Xu, Kemin; Craig, Maria Isabel; Shao, Hongxia; Ferreyra, Hebe; Rago, Maria Virginia; Romano, Marcelo; Uhart, Marcela; Sutton, Troy; Ferrero, Andrea; Perez, Daniel R.; Pereda, Ariel

2011-01-01

382

Molecular analysis of new isolates of Tomato leaf curl Philippines virus and an associated betasatellite occurring in the Philippines.  

PubMed

Three new begomovirus isolates and one betasatellite were obtained from a tomato plant exhibiting leaf curl symptom in Laguna, the Philippines. Typical begomovirus DNA components representing the three isolates (PH01, PH02 and PH03) were cloned, and their full-length sequences were determined to be 2754 to 2746 nucleotides. The genome organizations of these isolates were similar to those of other Old World monopartite begomoviruses. The sequence data indicated that PH01 and PH02 were variants of strain B of the species Tomato leaf curl Philippines virus, while PH03 was a variant of strain A of the species Tomato leaf curl Philippines virus. These isolates were designated ToLCPV-B[PH:Lag1:06], ToLCPV-B[PH:Lag2:06], and ToLCPV-A[PH:Lag3:06], respectively. Phylogenetic analysis revealed that the present isolates form a separate monophyletic cluster with indigenous begomoviruses reported earlier in the Philippines. A betasatellite isolated from same sample belongs to the betasatellite species Tomato leaf curl Philippines betasatellite and designated Tomato leaf curl Philippines betasatellite-[Philippines:Laguna1:2006], ToLCPHB-[PH:Lag1:06]. When co-inoculated with this betasatellite, tomato leaf curl Philippines virus induced severe symptoms in N. benthamiana and Solanum lycopersicum plants. Using a PVX-mediated transient assay, we found that the C4 and C2 proteins of tomato leaf curl Philippines virus and the ?C1 protein of ToLCPHB-[PH:Lag1:06] function as a suppressor of RNA silencing. PMID:21053032

Sharma, Pradeep; Matsuda, N; Bajet, N B; Ikegami, M

2011-02-01

383

Primary human immunodeficiency virus type 2 (HIV-2) isolates, like HIV-1 isolates, frequently use CCR5 but show promiscuity in coreceptor usage.  

PubMed

Coreceptor usage of primary human immunodeficiency virus type 1 (HIV-1) isolates varies according to biological phenotype. The chemokine receptors CCR5 and CXCR4 are the major coreceptors that, together with CD4, govern HIV-1 entry into cells. Since CXCR4 usage determines the biological phenotype for HIV-1 isolates and is more frequent in patients with immunodeficiency, it may serve as a marker for viral virulence. This possibility prompted us to study coreceptor usage by HIV-2, known to be less pathogenic than HIV-1. We tested 11 primary HIV-2 isolates for coreceptor usage in human cell lines: U87 glioma cells, stably expressing CD4 and the chemokine receptor CCR1, CCR2b, CCR3, CCR5, or CXCR4, and GHOST(3) osteosarcoma cells, coexpressing CD4 and CCR5, CXCR4, or the orphan receptor Bonzo or BOB. The indicator cells were infected by cocultivation with virus-producing peripheral blood mononuclear cells and by cell-free virus. Our results show that 10 of 11 HIV-2 isolates were able to efficiently use CCR5. In contrast, only two isolates, both from patients with advanced disease, used CXCR4 efficiently. These two isolates also promptly induced syncytia in MT-2 cells, a pattern described for HIV-1 isolates that use CXCR4. Unlike HIV-1, many of the HIV-2 isolates were promiscuous in their coreceptor usage in that they were able to use, apart from CCR5, one or more of the CCR1, CCR2b, CCR3, and BOB coreceptors. Another difference between HIV-1 and HIV-2 was that the ability to replicate in MT-2 cells appeared to be a general property of HIV-2 isolates. Based on BOB mRNA expression in MT-2 cells and the ability of our panel of HIV-2 isolates to use BOB, we suggest that HIV-2 can use BOB when entering MT-2 cells. The results indicate no obvious link between viral virulence and the ability to use a multitude of coreceptors. PMID:9971817

Mörner, A; Björndal, A; Albert, J; Kewalramani, V N; Littman, D R; Inoue, R; Thorstensson, R; Fenyö, E M; Björling, E

1999-03-01

384

Ancient Ancestry of KFDV and AHFV Revealed by Complete Genome Analyses of Viruses Isolated from Ticks and Mammalian Hosts  

PubMed Central

Background Alkhurma hemorrhagic fever virus (AHFV) and Kyasanur forest disease virus (KFDV) cause significant human disease and mortality in Saudi Arabia and India, respectively. Despite their distinct geographic ranges, AHFV and KFDV share a remarkably high sequence identity. Given its emergence decades after KFDV, AHFV has since been considered a variant of KFDV and thought to have arisen from an introduction of KFDV to Saudi Arabia from India. To gain a better understanding of the evolutionary history of AHFV and KFDV, we analyzed the full length genomes of 16 AHFV and 3 KFDV isolates. Methodology/Principal Findings Viral genomes were sequenced and compared to two AHFV sequences available in GenBank. Sequence analyses revealed higher genetic diversity within AHFVs isolated from ticks than human AHFV isolates. A Bayesian coalescent phylogenetic analysis demonstrated an ancient divergence of AHFV and KFDV of approximately 700 years ago. Conclusions/Significance The high sequence diversity within tick populations and the presence of competent tick vectors in the surrounding regions, coupled with the recent identification of AHFV in Egypt, indicate possible viral range expansion or a larger geographic range than previously thought. The divergence of AHFV from KFDV nearly 700 years ago suggests other AHFV/KFDV-like viruses might exist in the regions between Saudi Arabia and India. Given the human morbidity and mortality associated with these viruses, these results emphasize the importance of more focused study of these significant public health threats. PMID:21991403

Dodd, Kimberly A.; Bird, Brian H.; Khristova, Marina L.; Albarino, Cesar G.; Carroll, Serena A.; Comer, James A.; Erickson, Bobbie R.; Rollin, Pierre E.; Nichol, Stuart T.

2011-01-01

385

Characterization of Apricot pseudo-chlorotic leaf spot virus, A Novel Trichovirus Isolated from Stone Fruit Trees.  

PubMed

ABSTRACT A trichovirus closely related to Apple chlorotic leaf spot virus (ACLSV) was detected in symptomatic apricot and Japanese plum from Italy. The Sus2 isolate of this agent cross-reacted with anti-ACLSV polyclonal reagents but was not detected by broad-specificity anti- ACLSV monoclonal antibodies. It had particles with typical trichovirus morphology but, contrary to ACLSV, was unable to infect Chenopodium quinoa and C. amaranticolor. The sequence of its genome (7,494 nucleotides [nt], missing only approximately 30 to 40 nt of the 5' terminal sequence) and the partial sequence of another isolate were determined. The new virus has a genomic organization similar to that of ACLSV, with three open reading frames coding for a replication-associated protein (RNA-dependent RNA polymerase), a movement protein, and a capsid protein, respectively. However, it had only approximately 65 to 67% nucleotide identity with sequenced isolates of ACLSV. The differences in serology, host range, genome sequence, and phylogenetic reconstructions for all viral proteins support the idea that this agent should be considered a new virus, for which the name Apricot pseudo-chlorotic leaf spot virus (APCLSV) is proposed. APCLSV shows substantial sequence variability and has been recovered from various Prunus sources coming from seven countries, an indication that it is likely to have a wide geographical distribution. PMID:18943045

Liberti, D; Marais, A; Svanella-Dumas, L; Dulucq, M J; Alioto, D; Ragozzino, A; Rodoni, B; Candresse, T

2005-04-01

386

Phylogenetic relationships of the G gene sequence of bovine ephemeral fever virus isolated in Japan, Taiwan and Australia.  

PubMed

The G gene encoding the neutralization antigen of bovine ephemeral fever virus (BEFV) was characterized in order to define the virus's molecular epidemiology in Japan and the genetic relationships among the Japanese, Taiwanese and Australian isolates. The nucleotide and amino acid sequences of the gene were highly conserved among the Japanese strains, regardless of the year of isolation, and were closely related to the Taiwanese strains. By phylogenetic analysis, the Japanese and Taiwanese strains were classified clearly into three chronological clusters: 1966, 1984-1989 and 1996-2004, indicating that the epidemics of bovine ephemeral fever may occur almost simultaneously in both countries by the same genotype. On the other hand, the Australian strains were distantly related to these East Asian strains and placed in the independent fourth cluster of the phylogenetic tree. It is suggested that three amino acid substitutions at residues 224, 271 and 499 in the neutralizing epitopes, of which two generate new glycosylation sequences, are responsible for antigenic variations of bovine ephemeral fever virus. The cross-neutralization test using the bovine ephemeral fever virus isolated in Japan demonstrated that the vaccine developed based on the oldest Japanese strain, YHL, appears to still be effective for controlling bovine ephemeral fever in Japan. PMID:19201554

Kato, Tomoko; Aizawa, Maki; Takayoshi, Katsunori; Kokuba, Tamotsu; Yanase, Tohru; Shirafuji, Hiroaki; Tsuda, Tomoyuki; Yamakawa, Makoto

2009-06-12

387

Characterization of Hepatitis C Virus Genotypes by Direct Sequencing of HCV 5?UTR Region of Isolates from Saudi Arabia  

PubMed Central

The current study was designed to determine the Hepatitis C Virus (HCV) genotypes in a representative sample of HCV chronically infected patients in Saudi Arabia. All HCV isolates were genotyped by sequencing of the 5?UTR region and newly identified HCV isolates were identified. Specific universal primers targeting 5?UTR region were used for both amplification and sequencing of all isolates that resulted in 244 bp fragment which represent about 80% of 5?UTR region. Most of HCV isolates in this study were genotype 4 (76.4%) where only few isolates were recognized as genotype 1 (19.6%). All results were compared to HCV reference sequences from LOS ALAMOS HCV database, considering only the complete full genomes for the main phylogenetic analysis. Sequences that showed maximum identity (98% –100%) were selected. Most isolates were identical with HCV genotype 4 references. Some isolates were similar to different subtypes of HCV genotypes 4, 1 and 6. Phylogenetic analysis showed resemblance of most isolates to similar ones from the Far East, North America and Egypt. Using sequence Weblogo, Alignment analysis of isolated HCV genotypes 4 and 1 showed 92% and 95.5% nucleotide conservation, respectively. There was no predominant nucleotide in the varied sites, in both genotypes. All isolated sequences were submitted to GenBank database. PMID:25099694

Shier, Medhat K.; El-Wetidy, Mohammad S.; Ali, Hebatallah H.; Al-Qattan, Mohammad M.

2014-01-01

388

Langerhans cell tropism of human immunodeficiency virus type 1 subtype A through F isolates derived from different transmission groups.  

PubMed Central

To test the hypothesis that some subtypes of human immunodeficiency virus type 1 (HIV-1), especially subtype E, are more likely to infect mature Langerhans cells (mLC), we titrated a panel of 26 primary HIV-1 isolates of subtypes A through F on peripheral blood mononuclear cells (PBMC) and mLC. The majority of HIV-1 isolates from heterosexually infected patients did not show a preferred tropism for mLC compared to homosexually transmitted HIV-1 isolates. Only 6 of 26 isolates, 2 from patients infected by homosexual contact and 4 from patients infected by heterosexual contact, showed a higher infectivity for mLC than for PBMC. Both syncytium-inducing and non-syncytium-inducing isolates were able to infect mLC which express mRNA for the chemokine receptors CCR3, CCR5, and CXCR4. PMID:9311896

Dittmar, M T; Simmons, G; Hibbitts, S; O'Hare, M; Louisirirotchanakul, S; Beddows, S; Weber, J; Clapham, P R; Weiss, R A

1997-01-01

389

Comparative analysis of the genomes of two isolates of cowpea aphid-borne mosaic virus (CABMV) obtained from different hosts.  

PubMed

The complete genomic sequences of two cowpea aphid-borne mosaic virus (CABMV) isolates from Brazil, MG-Avr from passion fruit (which also infects cowpea), and BR1 from peanut (which also infects cowpea, but not passion fruit), were determined. Their nucleotide sequences are 89% identical and display 85% identity to that of CABMV-Z. Both isolates have the typical potyvirus genome features. P3 and VPg are the most conserved proteins, with 99% amino acid sequence identity between the two isolates, and P1 is the most variable, with 50% identity. A significant variation exists at the 5'-end of the genome between the Brazilian isolates and CABMV-Z. However, this variation does not correlate with the biological properties of these three isolates. PMID:21409445

Barros, Danielle R; Alfenas-Zerbini, Poliane; Beserra, José Evando A; Antunes, Tathiana F S; Zerbini, F Murilo

2011-06-01

390

Effect of temperature on peroxidase activity, isozyme patterns and concentration of threadlike particles in tristeza—infected citrus plants  

Microsoft Academic Search

Peroxidase activity (PA) in various tristeza-infected citrus varieties was significantly higher than in healthy controls.\\u000a In leaves and bark of Egyptian sour lime and Palestine sweet lime, PA was correlated with symptom severity and content of\\u000a threadlike particles (TLP). In infected roots of Egyptian sour lime, there was an increase in PA but TLP content was minimal.\\u000a \\u000a Temperatures above 22°C

M. Bar-Joseph; G. Loebenstein

1973-01-01

391

Pandemic swine-origin H1N1 influenza A virus isolates show heterogeneous virulence in macaques.  

PubMed

The first influenza pandemic of the new millennium was caused by a newly emerged swine-origin influenza virus (SOIV) (H1N1). This new virus is characterized by a previously unknown constellation of gene segments derived from North American and Eurasian swine lineages and the absence of common markers predictive of human adaptation. Overall, human infections appeared to be mild, but an alarming number of young individuals presented with symptoms atypical for seasonal influenza. The new SOIV also showed a sustained human-to-human transmissibility and higher reproduction ratio than common seasonal viruses, altogether indicating a higher pathogenic potential for this newly emerged virus. To study the virulence of the SOIV, we used a recently established cynomolgus macaque model and compared parameters of clinical disease, virology, host responses, and pathology/histopathology with a current seasonal H1N1 virus. We here show that infection of macaques with two genetically similar but clinically distinct SOIV isolates from the early stage of the pandemic (A/Mexico/4108/2009 and A/Mexico/InDRE4487/2009) resulted in upper and lower respiratory tract infections and clinical disease ranging from mild to severe pneumonia that was clearly advanced over the mild infection caused by A/Kawasaki/UTK-4/2009, a current seasonal strain. Unexpectedly, we observed heterogeneity among the two SOIV isolates in virus replication, host transcriptional and cytokine responses, and disease progression, demonstrating a higher pathogenic potential for A/Mexico/InDRE4487/2009. Differences in virulence may explain more severe disease, as was seen with certain individuals infected with the emerged pandemic influenza virus. Thus, the nonhuman primate model closely mimics influenza in humans. PMID:21084481

Safronetz, David; Rockx, Barry; Feldmann, Friederike; Belisle, Sarah E; Palermo, Robert E; Brining, Douglas; Gardner, Don; Proll, Sean C; Marzi, Andrea; Tsuda, Yoshimi; Lacasse, Rachel A; Kercher, Lisa; York, Anthony; Korth, Marcus J; Long, Dan; Rosenke, Rebecca; Shupert, W Lesley; Aranda, Celia Alpuche; Mattoon, John S; Kobasa, Darwyn; Kobinger, Gary; Li, Yan; Taubenberger, Jeffery K; Richt, Jürgen A; Parnell, Michael; Ebihara, Hideki; Kawaoka, Yoshihiro; Katze, Michael G; Feldmann, Heinz

2011-02-01

392

Molecular Characterization and Variation of the Broad bean wilt virus 2 Isolates Based on Analyses of Complete Genome Sequences  

PubMed Central

The full-genome sequences of fourteen isolates of Broad bean wilt virus 2 (BBWV2), collected from broad bean, pea, spinach, bell pepper and paprika plants in Korea during the years 2006–2012, were determined and analyzed comparatively along with fifteen previously reported BBWV2 genome sequences. Sequence analyses showed that RNA-1 and RNA-2 sequences of BBWV2 Korean isolates consisted of 5950–5956 and 3568–3604 nucleotides, respectively. Full-length genome sequence-based phylogenetic analyses revealed that the BBWV2 Korean isolates could be divided into three major groups comprising GS-I (isolates BB2 and RP7) along with isolate IP, GS-II (isolates BB5, P2, P3 and RP3) along with isolate B935, and GS-III including 16 BBWV2 Korean isolates. Interestingly, GS-III appears to be newly emerged and predominant in Korea. Recombination analyses identified two recombination events in the analyzed BBWV2 population: one in the RNA-1 of isolate K and another one in the RNA-2 of isolate XJ14-3. However, no recombination events were detected in the other 21 Korean isolates. On the other hand, out of 29 BBWV2 isolates, 16 isolates were found to be reassortants, of which each RNA segment (i.e. RNA1 and RNA2) was originated from different parental isolates. Our findings suggested that reassortment rather than recombination is a major evolutionary force in the genetic diversification of BBWV population in Korea. PMID:25288968

Kwak, Hae-Ryun; Kim, Mi-Kyeong; Lee, Ye-Ji; Seo, Jang-Kyun; Kim, Jeong-Soo; Kim, Kook-Hyung; Cha, Byeongjin; Choi, Hong-Soo

2013-01-01

393

Isolation and Characterization of Highly Replicable Hepatitis C Virus Genotype 1a Strain HCV-RMT  

PubMed Central

Multiple genotype 1a clones have been reported, including the very first hepatitis C virus (HCV) clone called H77. The replication ability of some of these clones has been confirmed in vitro and in vivo, although this ability is somehow compromised. We now report a newly isolated genotype 1a clone, designated HCV-RMT, which has the ability to replicate efficiently in patients, chimeric mice with humanized liver, and cultured cells. An authentic subgenomic replicon cell line was established from the HCV-RMT sequence with spontaneous introduction of three adaptive mutations, which were later confirmed to be responsible for efficient replication in HuH-7 cells as both subgenomic replicon RNA and viral genome RNA. Following transfection, the HCV-RMT RNA genome with three adaptive mutations was maintained for more than 2 months in HuH-7 cells. One clone selected from the transfected cells had a high copy number, and its supernatant could infect naďve HuH-7 cells. Direct injection of wild-type HCV-RMT RNA into the liver of chimeric mice with humanized liver resulted in vigorous replication, similar to inoculation with the parental patient’s serum. A study of virus replication using HCV-RMT derivatives with various combinations of adaptive mutations revealed a clear inversely proportional relationship between in vitro and in vivo replication abilities. Thus, we suggest that HCV-RMT and its derivatives are important tools for HCV genotype 1a research and for determining the mechanism of HCV replication in vitro and in vivo. PMID:24358200

Arai, Masaaki; Tokunaga, Yuko; Takagi, Asako; Tobita, Yoshimi; Hirata, Yuichi; Ishida, Yuji; Tateno, Chise; Kohara, Michinori

2013-01-01

394

Geographically distant isolates of the crinivirus Cucurbit yellow stunting disorder virus show very low genetic diversity in the coat protein gene  

Microsoft Academic Search

The population structure and genetic variation of Cucurbit yellow stunting disorder virus (CYSDV) isolates were estimated by single-strand confor- mation polymorphism and nucleotide sequence analyses of the CYSDV coat protein gene. Analysis of 71 isolates collected from Spain, Jordan, Turkey, Lebanon, Saudi Arabia and North America showed that, from a genetic viewpoint, these isolates could be divided into two diverged

Luis Rubio; Yusuf Abou-Jawdah; Han-Xin Lin; Bryce W. Falk

2001-01-01

395

Complete genome sequence and pathogenesis of bovine viral diarrhea virus JL-1 isolate from cattle in China  

PubMed Central

Background Bovine viral diarrhea virus (BVDV) is a pathogen found worldwide in calves. It can cause significant economic losses in agriculture. Many BVDV strains have been isolated in China. However, the pathogenesis and complete gene characteristics of BVDV isolate have yet not been reported in China. Here, a BVDV isolate was isolated and its pathogenesis and complete genome were studied. Results A new isolate of bovine viral diarrhea virus, named JL-1, was isolated from the spleen of a sick cow with diarrhea using MDBK cell culture. The complete genome of JL-1 is 12,276 nucleotides and contains a 5?-UTR of 382 nucleotides, a 3?-UTR of 188 nucleotides, and a large ORF encoding a polyprotein consisting of 3,901 amino acids. Genomic comparison and phylogenetic analyses of complete genomic sequence clearly showed that JL-1 fell into the BVDV-1b subtype. The result of pathogenesis of JL-1 strain showed that all infected calves developed clinical signs of elevated rectal temperatures, decreased leucopenia, and viral discharge. Viral antigen was detected in infected animal tissues using immunohistochemistry. Animals in the mock were normal. These results demonstrated that BVDV JL-1 was a virulent strain. Conclusions This is the first study to report the pathogenesis and complete gene characterization of the BVDV strain in China. This report may set a good foundation for further study of BVDV in China. PMID:24708732

2014-01-01

396

Isolation of highly fusogenic variants of simian virus 5 from persistently infected cells that produce and respond to interferon.  

PubMed Central

A series of experiments were undertaken to examine how interferon and neutralizing antibodies influence the ability of simian virus 5 (SV5) (strain W3) to establish and maintain persistent infections in murine cells. In contrast to the rapid decline in SV5 protein synthesis observed in murine BALB/c fibroblasts (BF cells), which produce and respond to interferon, between 24 and 48 h postinfection there was no inhibition of virus protein synthesis in MSFI- cells, skin fibroblasts derived from alpha/beta-interferon receptor knockout BALB/c mice. Furthermore, the addition of anti-interferon antibodies to the culture medium of infected BF cells significantly reduced the observed decline in virus protein synthesis. Following infection of untreated BF cells, the majority replicated virus but survived the infection and eventually cleared the virus after 8 to 15 days. However, not all the cells were cured, and the cultures became persistently infected. Upon passage of persistently infected cultures, the virus fluxed between active and repressed states as a consequence of interferon production. This resulted in a balance being reached in which only 5 to 20% of the cells were infected at any one time. After 30 passages of the persistently infected cells, highly fusogenic virus variants arose (one of which was isolated and termed W3-f). W3-f remained as sensitive to interferon as the parental W3 isolate but, in the absence of interferon, spread much more rapidly than the parental W3 strain through BF cell monolayers. Sequence analysis revealed no deduced amino acid differences between the F proteins of W3 and W3-f. BF cell cultures persistently infected with W3-f were rapidly cleared of virus by the addition of virus-neutralizing antibodies to the culture medium. In contrast, neutralizing antibodies had little effect on the numbers of cells persistently infected with W3 over several passages. These results suggest that the ability of paramyxoviruses to cause cell-cell fusion may be selected for in vivo as a consequence of their adaptation to the interferon response rather than their need to escape from neutralizing antibodies. The significance of these observations with regard to persistent parainfluenza virus infections in vivo is further discussed. PMID:9371592

Young, D F; Didcock, L; Randall, R E

1997-01-01

397