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Sample records for tristeza virus isolate

  1. Molecular Characterization of Citrus tristeza virus Isolates from Panama

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Twelve isolates of Citrus tristeza virus (CTV) were collected from the main citrus growing regions in Panama and characterized at the molecular level. The CTV coat protein gene (CPG) was amplified by RT-PCR, and the amplified PCR products were cloned and sequenced. The sequences analyses showed the ...

  2. Molecular Characterization of Citrus Tristeza Virus Isolates from Mild Strain Cross Protection Experiments in Peru

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seven isolates of Citrus tristeza virus (CTV) collected from the Topara Nursery, Lima, Peru were established as in planta cultures in Madam Vinous sweet orange in the Exotic Citrus Pathogen Quarantine Greenhouse, Beltsville, MD. Two of the isolates were collected from non-cross protected plants; Pe...

  3. The prevalence of the citrus tristeza virus trifoliate resistant breaking genotype among Puerto Rican isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) isolates have been grouped previously into five genotypes: T3, T30, T36, VT, B165 based on symptoms, host range and genomic sequence data. A sixth genotype has recently been identified with the novel property of replicating in trifoliate orange trees, a non host for the o...

  4. Molecular characterization of Peruvian Citrus tristeza virus isolates based on 3’UTR sequences

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus in Peru was decimated by quick decline and stem pitting strains of Citrus tristeza virus (CTV). Commercial citrus production in Peru is being restored by use of CTV cross-protection. To characterize the predominant CTV strains involved, Peruvian CTV isolates from “protected” and “non-protecti...

  5. CPm gene diversity in field isolates of Citrus tristeza virus from Colombia.

    PubMed

    Oliveros-Garay, Oscar Arturo; Martinez-Salazar, Natalhie; Torres-Ruiz, Yanneth; Acosta, Orlando

    2009-01-01

    The nucleotide sequence diversity of the CPm gene from 28 field isolates of Citrus tristeza virus (CTV) was assessed by SSCP and sequence analyses. These isolates showed two major shared haplotypes, which differed in distribution: A1 was the major haplotype in 23 isolates from different geographic regions, whereas R1 was found in isolates from a discrete region. Phylogenetic reconstruction clustered A1 within an independent group, while R1 was grouped with mild isolates T30 from Florida and T385 from Spain. Some isolates contained several minor haplotypes, which were very similar to, and associated with, the major haplotype. PMID:19882104

  6. Population structure and genetic diversity within California Citrus tristeza virus (CTV) isolates.

    PubMed

    Kong, P; Rubio, L; Polek, M; Falk, B W

    2000-10-01

    The Closterovirus, Citrus tristeza virus (CTV) is an aphid-borne RNA virus that is the causal agent of important worldwide economic losses in citrus. Biological and molecular variation has been observed for many CTV isolates. In this work we detected and analyzed sequence variants (haplotypes) within individual CTV isolates. We studied the population structure of five California CTV isolates by single strand conformation polymorphism (SSCP) analysis of four CTV genomic regions. Also, we estimated the genetic diversity within and between isolates by analysis of haplotype nucleotide sequences. Most CTV isolates were composed of a population of genetically related variants (haplotypes), one being predominant. However in one case, we found a high nucleotide divergence between haplotypes of the same isolate. Comparison of these haplotypes with those from other isolates suggests that some CTV isolates could have arisen as result of a mixed infection of two divergent isolates. PMID:11129629

  7. Molecular analyses revealed genetic complexity in Citrus tristeza virus Dekopon isolate and its aphid-transmitted progeny

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An assessment was made of the disease potential of a Citrus tristeza virus (CTV) isolate designated Dekopon found in a hybrid mandarin variety topworked in a citrus planting in Fresno County, CA. After aphid transmissions (AT), parental and AT isolates were analyzed by SSCP, genotyping with multipl...

  8. Complete Nucleotide Sequence of a New Genotype of Citrus Tristeza Virus from an Isolate Having a Mixed Infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An isolate of Citrus tristeza virus (CTV) that causes severe stem pitting in grapefruits (# 3800) was used for sequencing. Analysis of the isolate revealed the presence of at least three different populations, one belonging to T30 genotype and the other two belonging to new genotypes, designated T2K...

  9. Assessment of the Citrus tristeza virus isolates detected in spring 2007 at the Lindcove Research and Extension Center, Exeter, California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus was detected in at least 50 trees at the 71 ha Lindcove Research and Extension Center (LREC) near Exeter, Calif. in spring 2007. The purpose of this research was to assess genetic diversity and aphid transmissibility of these isolates. Nine representative trees were sampled o...

  10. Molecular characterization of Cirus tristeza virus isolates associated with stem pitting CTV cross-protection in Peru

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During the 1970s and early 1980s, the Peruvian citrus industry was destroyed by severe Citrus tristeza virus (CTV) strains spread by the brown citrus aphid. The Topara Nursery, located 180 km south of Lima Peru, selected and identified CTV isolates that confer cross-protection against virulent stem...

  11. Citrus tristeza virus-host interactions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 s...

  12. Aphid Transmission Alters the Genomic and Defective RNA Populations of Citrus tristeza virus Isolates.

    PubMed

    Albiach-Martí, M R; Guerri, J; de Mendoza, A H; Laigret, F; Ballester-Olmos, J F; Moreno, P

    2000-02-01

    ABSTRACT A total of 14 Spanish isolates of Citrus tristeza virus (CTV) and 1 isolate from Japan were transmitted by Aphis gossypii, and the subisolates obtained were compared with the source isolates for symptom expression and double-stranded RNA (dsRNA) pattern. Of the 14 Spanish isolates, 9 showed altered dsRNA patterns after aphid transmission but only minor variations in the intensity of symptoms induced on Mexican lime. Northern blot hybridization with complementary DNA (cDNA) probes corresponding to both the 5' and the 3' termini of the CTV genomic RNA (gRNA) showed that the dsRNA bands that could be used to discriminate between the dsRNA pattern of the source and the aphid-transmitted isolates were the replicative forms of defective RNAs (D-RNAs). Conversely, the Japanese isolate and two subisolates obtained from it by aphid transmission had the same dsRNA pattern, but one of the subisolates induced milder symptoms in several hosts. Dot-blot hybridization with cDNA probes representing several regions of the gRNA showed that most of the aphid-transmitted isolates differed from the corresponding source isolate by their hybridization pattern. Our results indicate that aphid transmission often sorts the populations of gRNA variants and D-RNAs present in CTV isolates. PMID:18944601

  13. Molecular Marker Analysis of Citrus tristeza virus (CTV) isolates from the Dominican Republic

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Samples of citrus tissue infected with Citrus tristeza virus (CTV) were collected from Persian lime, mandarin, Washington navel, Valencia or grapefruit trees from various locations in the Dominican Republic. Desiccated tissue samples were re-hydrated and virions extracted by grinding samples in buff...

  14. Real-time RT-PCR Assay for Detection and Differentiation of Citrus Tristeza Virus Isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Multiplex one step real time RT-PCR assays using TaqMan probes were developed for detection and strain differentiation of Citrus tristeza virus (CTV). For broad spectrum CTV detection, a TaqMan primer and Cy5-labeled probe were designed using CP gene sequences. An internal control was developed us...

  15. Genetic diversity of Citrus tristeza virus isolates spreading in Central California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A rapid increase in the number of trees infected with Citrus tristeza virus (CTV) was observed in several locations in Tulare, County, CA during 2007. Since trees had been tested annually, these infections represent new infections. Leaf and bark tissue were sampled from infected trees and used for...

  16. Real-time RT-PCR assay for detection and differentiation of Citrus tristeza virus isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    For universal detection of Citrus tristeza virus (CTV) strains by real time RT-PCR, a protocol was developed based on a set of primers and a Cy5-labeled TaqMan probe. This test included primers and a TET-labeled TaqMan probe selected on the mitochondrial nad5 gene for the simultaneous detection of ...

  17. HISTOLOGY OF SWEET ORANGE STEM PITTING CAUSED BY AN AUSTRALIAN ISOLATE OF CITRUS TRISTEZA VIRUS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Some strains of the citrus tristeza virus (CTV) cause stem pitting in sweet orange (Citrus sinensis (L.) Osbeck). This abnormality causes tree decline and reduction in fruit size and yield of affected citrus trees. Stem-pitting symptoms can occur on trunks, on all sizes of limbs, and on the twigs ...

  18. Genetic Diversity of Citrus tristeza Virus Isolates Collected Recently in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Surveys conducted over the past several years show a dramatic increase in Citrus tristeza virus (CTV) incidence in several locations in Central California. Our objective was to assess genetic diversity of current CTV field populations and determine their phylogenetic relationships with representati...

  19. Characterization of Citrus Tristeza Virus Isolates by Single-strand Conformation Polymorphism Analysis of the Coat Protein Gene

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A method is needed to rapidly assess Citrus tristeza virus (CTV) strains and to identify mixed populations in tristeza-infected trees. Single-strand conformation polymorphism (SSCP) can detect point mutations in DNA fragments and determine the structure of viral populations. Previous reports utili...

  20. Nucleotide heterogeneity at the genomic 5’- and 3’-termini of California (CA) isolates of Citrus tristeza virus (CTV)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Nucleotide (nt) sequences in the genomic ends of sense (+)-RNA viruses serve essential biological functions and are important considerations in the construction of infectious clones. Two isolates of Citrus tristeza virus (CTV) from California (CA) having a T30- and a T36-genotype were inoculated in ...

  1. Complete 3' end genome analysis of the asymptomatic citrus tristeza virus isolate B192 and its eight single aphid transmitted subisolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The most important viral disease of citrus is caused by Citrus tristeza virus (CTV). CTV infection often exists in field isolates as a complex of multiple genotypes. Aphid transmission is important for CTV dispersal. The complete 3' terminal half sequences of the asymptomatic CTV isolate B192 and it...

  2. Characterization of a novel citrus tristeza virus genotype within three cross-protecting source GFMS12 sub-isolates in South Africa by means of Illumina sequencing.

    PubMed

    Zablocki, Olivier; Pietersen, Gerhard

    2014-08-01

    Tristeza disease (caused by citrus tristeza virus, CTV) is currently controlled in South Africa by means of cross-protection. In this study, we characterized the CTV populations of three grapefruit mild strain 12 (GFMS12) single-aphid-transmission-derived sub-isolates at the whole-genome level using Illumina sequencing technology. A novel South African isolate (CT-ZA3, of the T68 genotype) was shown to be the dominant genotype in all GFMS12 sub-isolates tested, along with reads unique to various other genotypes occurring as minor components. Uncertainty remains as to the significance of these minor components. PMID:24623089

  3. Molecular Analysis Among MCA13-reactive Isolates Reveals a Strategy for Rapid Assessment of Citrus tristeza Virus Severity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) genotypes vary in disease severity ranging from symptomless to virulent (stem pitting) in commercial citrus plantings. Because CTV is spread by propagation and by aphid vectors, rapid identification and virulence typing are critical for control and interdiction activitie...

  4. Development and application of a hexaplex reverse transcription polymerase chain reaction for screening global citrus tristeza virus isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The discovery of the diversity of Citrus tristeza virus (CTV) genotypes has complicated detection and diagnostic measures. To simplify the identification and differentiation of CTV genotypes, an efficient multiplex reverse transcription polymerase chain reaction (M-RT-PCR) technique for the screenin...

  5. Molecular analysis among MCA13-reactive isolates reveals a rapid strategy for assessment of Citrus tristeza virus severity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) usually occurs as a complex of strains that vary greatly in severity and aphid transmissibility. A rapid assay, therefore, is needed to distinguish potentially mild vs. severe strains of CTV for disease mitigation. An economical and practical strategy to screen for poten...

  6. Survey of citrus tristeza virus populations in Central California that react with MCA13 monoclonal antibody

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Citrus Pest Detection Program (CPDP) of the Central California Tristeza Eradication Agency monitors Citrus tristeza virus (CTV) in Central California. MCA13 is a severe strain discriminating monoclonal antibody used to screen for potentially virulent CTV isolates. MCA13-reactive CTV isolates are...

  7. Molecular diversity of Citrus tristeza virus in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is a serious citrus pathogen worldwide. Recent genetic studies have identified five standard CTV genotypic groups: T30, VT, T36, T3, and B165/T68. Field surveys performed in California in 2008-2010 identified primarily MCA13-negative CTV isolates with T30-like genotype. C...

  8. Deep sequencing and analysis of small RNAs in sweet orange grafted on sour orange infected with two citrus tristeza virus isolates prevalent in Sicily.

    PubMed

    Licciardello, Grazia; Scuderi, Giuseppe; Ferraro, Rosario; Giampetruzzi, Annalisa; Russo, Marcella; Lombardo, Alessandro; Raspagliesi, Domenico; Bar-Joseph, Moshe; Catara, Antonino

    2015-10-01

    Two representative isolates of a citrus tristeza virus population in Sicily, SG29 (aggressive) and Bau282 (mild), were sequenced via viral small RNAs (vsRNA) produced in budlings of sweet orange grafted on sour orange. Phylogenetic relationships with Mediterranean and exotic isolates revealed that SG29 clustered within the "VT-Asian" subtype, whereas Bau282 belonged to the cluster T30. The study confirms that molecular data need to be integrated with bio-indexing in order to obtain adequate information for risk assessment. PMID:26175068

  9. Citrus tristeza virus-aphid interactions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A review chapter on aphid transmission of Citrus tristeza virus is provided for a book on “Vector-Mediated Transmission of Plant Pathogens”. Earliest uses of citrus goes back over two millennia as items of trade, gifts and medicinal compounds. Citrus propagation during this period was by seed and si...

  10. Genetic Marker Analysis of a Global Collection of Isolates of Citrus tristeza virus: Characterization and Distribution of CTV Genotypes and Association with Symptoms.

    PubMed

    Hilf, Mark E; Mavrodieva, Vessela A; Garnsey, Stephen M

    2005-08-01

    ABSTRACT Genetic markers amplified from three noncontiguous regions by sequence specific primers designed from the partial or complete genome sequences of Citrus tristeza virus (CTV) isolates T3, T30, T36, and VT were used to assess genetic relatedness of 372 isolates in an international collection. Eighty-five isolates were judged similar to the T3 isolate, 81 to T30, 11 to T36, and 89 to VT. Fifty-one isolates were mixed infections by two or more identifiable viral genotypes, and 55 isolates could not be assigned unequivocally to a group defined by marker patterns. Maximum parsimony analysis of aligned marker sequences supported the grouping of isolates on the basis of marker patterns only. Specific disease symptoms induced in select citrus host plants were shared across molecular groups, although symptoms were least severe among isolates grouped by markers with the T30 isolate and were most severe among isolates grouped by markers with the T3 isolate. Isolates assigned the same genotype showed variable symptoms and symptom severity. A classification strategy for CTV isolates is proposed that combines genetic marker patterns and nucleotide sequence data. PMID:18944413

  11. Polymorphism of the 5' terminal region of Citrus tristeza virus (CTV) RNA: incidence of three sequence types in isolates of different origin and pathogenicity.

    PubMed

    Ayllón, M A; López, C; Navas-Castillo, J; Garnsey, S M; Guerri, J; Flores, R; Moreno, P

    2001-01-01

    Sequences of the 5' terminal region of the genomic RNA from eight isolates of Citrus tristeza virus (CTV) were previously classified into three types (I, II and III), with intragroup sequence identity higher than 88% and intergroup sequence identity as low as 44%. Sequencing of an additional 58 cDNA clones from 15 virus isolates showed that all sequences could be unequivocally assigned to one of the three types previously established. The relative frequency of each sequence type was assessed in 57 CTV isolates of different geographic origin and pathogenic characteristics by RT-PCR with sets of type-specific primers using CTV dsRNA as template. None of the isolates yielded amplification of the type I or II sequences alone, but in 19 of them type III sequences were the only amplification product detected. Within isolates containing more than one sequence type, eight had type II and III sequences, 11 had type I and III sequences, and 19 had sequences of the three types. Isolates containing only type III sequences caused only mild to moderate symptoms in Mexican lime, an indicator species for most CTV isolates, whereas isolates causing stem pitting in sweet orange an/or grapefruit, generally contained sequences type II. None of the sequence types could be traced to a precise geographic area, as all types were detected in isolates from at least nine of the 12 countries from which samples were taken. PMID:11266215

  12. Cross-protection by Citrus Tristeza Virus: prevention of superinfection occurs only when plants are protected by isolates of the same strain.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is the most important virus of citrus worldwide. CTV has a 19.3 kb RNA genome, is transmitted by aphids, and accumulates in trees predominantly in phloem-associated cells. Inoculation of trees with mild strains of CTV has been used to allow citrus production in areas wher...

  13. Evidence of Recombinant Citrus tristeza virus Isolate Occurring in Acid Lime cv. Pant Lemon Orchard in Uttarakhand Terai Region of Northern Himalaya in India.

    PubMed

    Singh, Jaywant Kumar; Tarafdar, Avijit; Sharma, Susheel Kumar; Biswas, Kajal Kumar

    2013-06-01

    The present study for the first time describes biological and molecular characterization of Citrus tristeza virus (CTV) occurring in the Terai area of Uttarakhand State in Northern Himalaya region of India. Direct antigen coated-ELISA and reverse transcriptase-polymerase chain reaction (RT-PCR) detected the CTV infection in Acid lime cv. Pant lemon (Citrus aurantifolia) orchards of Pantnagar with an estimated disease incidence of 16.6-20.5 %. To know the biological and genetic properties, an isolate, CTV Pant 4 was characterized. Isolate Pant 4 could be graft transmitted to Kinnow, Nagpur and Darjeeling mandarins, Mosambi sweet orange, Kagzi lime, Sweet lime, Sour orange but not to Rough lemon. The sequence analyses of the 5'ORF1a (3038 nucleotides) of LPro domain and 3'end (2058 nt) covering ORF7-ORF10 regions of the CTV genome revealed that Pant 4 was closely related to the previously reported Indian CTV isolate, Kpg3 from Northeastern Himalaya region with 97 and 98 % sequence identity, respectively. Whereas, it differed from the previously reported CTV isolate B165 from Southern India with 79 and 92 % identity, respectively for 5'ORF1a and 3' end regions. Recombination and SplitsTree decomposition analyses indicated that CTV isolate Pant 4 was a recombinant isolate originating from Kpg3 as a major and B165 as a minor donor. PMID:24426255

  14. Stem pitting Citrus tristeza virus predominantly transmitted by the brown citrus aphid from mixed infections containing non-stem pitting and stem pitting isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is a phloem-limited closterovirus that produces a variety of symptoms in various Citrus spp. One of these symptoms is stem pitting (SP). SP does not occur in all Citrus spp. but when it does it may cause low tree vigor, decline and an economically-significant reduction ...

  15. Rapid Assessment of the Citrus Tristeza Virus Isolates Detected in Spring 2007 at the Lindcove Research and Extension Center, Exeter, Calif.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) was detected in at least 50 trees at the 71 ha Lindcove Research and Extension Center (LREC) near Exeter, Calif. in spring 2007. In the previous 3 years, 3, 1, and 5 trees were infected. The purpose of this research was to assess the aphid transmissibility and molecular ...

  16. Deep sequencing of viral small-RNAs of citrus tristeza virus (CTV) reveals genomic differences between two Italian isolates of CTV

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A recent Citrus tristeza virus (CTV) epidemic of quick decline (QD) killed many sweet orange trees grafted on sour orange rootstock in Sicily but left some asymptomatic trees in the same field. Recent reports indicated cross-protection involves exclusion of a severe CTV strain by a mild strain of th...

  17. Novel mild strains of Citrus tristeza virus from California and Peru.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) has caused great economic losses to citrus worldwide. CTV isolates from California were identified which reacted to MCA13 but were mild in biological indexing tests. Molecular markers were developed to differentiate these isolates from established CTV genotypes and the is...

  18. Distribution, genetic diversity and recombination analysis of Citrus tristeza virus of India

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) isolates representing all the citrus growing geographical zones of India were analyzed for sequence of the 5'ORF1a fragments of the partial LProI domain and for the coat protein (CP) gene. The sequences were compared with previously reported Indian and CTV genotypes from...

  19. Genetic differentiation and biology of Citrus tristeza virus populations spreading in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) isolates were collected from more than 1500 trees in citrus groves in Tulare, Kern, Ventura, Riverside and San Diego Counties for laboratory tests to assess molecular and biological properties of CTV strains currently in California. Tests included serology with MCA13 mon...

  20. Dramatic Change in Citrus tristeza virus populations in the Dominican Republic

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is the most destructive viral pathogen of citrus and has been an important concern for the citrus industry in the Dominican Republic. Earlier studies documented widespread distribution of mild isolates of the T30 genotype, which caused no disease in the infected trees, an...

  1. Production of Polyclonal Antibodies to the Recombinant Coat Protein of Citrus tristeza virus and Their Effectiveness for Virus Detection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The p25 coat protein gene of three Citrus tristeza virus (CTV) isolates, two from Mexico and one from India, was amplified by RT-PCR and further cloned and expressed in Escherichia coli cells. The recombinant coat protein (rCP) of the three CTV isolates was injected into rabbits and goats for antibo...

  2. Transgenic Resistance to Citrus tristeza virus in Grapefruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Grapefruit (Citrus paradisi) transgenic plants transformed with a variety of constructs derived from the Citrus tristeza virus (CTV) genome were tested for their resistance to the virus. Most transgenic lines were susceptible (27 lines), a few were partially resistant (6 lines) and only one line, tr...

  3. Current status of Citrus tristeza virus in Central California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Lindcove Research and Extension Center (LREC), Exeter, CA has 51 ha of citrus and is the field site and screenhouses for the University of California Citrus Clonal Protection Program (CCPP). LREC maintains a zero tolerance of Citrus tristeza virus (CTV) infected trees to protect the CCPP and re...

  4. Thirty years of citrus tristeza virus observations in Peru

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Peruvian citrus industry was devastated by epidemics of Citrus tristeza virus (CTV) decline (CTV-D) on sour orange rootstock between 1950 and 1965 and CTV stem pitting (SP) between 1965 and 1985. CTV-SP debilitates citrus and fruit production regardless of rootstock. Control of CTV-SP by mild st...

  5. Emergence and Phylodynamics of Citrus tristeza virus in Sicily, Italy

    PubMed Central

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

    2013-01-01

    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

  6. Genetic differentiation and biology of Citrus tristeza virus populations spreading in eradicative and non-eradicative areas of California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previous studies showed Citrus tristeza virus (CTV) isolates collected from the 1970’s in California were closely related to the mild T30 isolate; only a few severe strains such as SY568 (Riverside) and Dekopon (Orange Cove) were found and subsequently eradicated. CTV is now spreading rapidly in so...

  7. Citrus tristeza virus: Evolution of Complex and Varied Genotypic Groups

    PubMed Central

    Harper, S. J.

    2013-01-01

    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

  8. Use of the Coat Protein (CP) and minor CP Intergene Sequence to Discriminate Severe Strains of Citrus tristeza virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A rapid assay is a needed to differentiate mild vs severe strains of Citrus tristeza virus (CTV). Multiple alignment performed on the coat protein (CP) and the minor coat protein (CPm) intergene sequences (~80-100 bp) from different CTV isolates revealed that severe strains generally associated wit...

  9. Citrus tristeza virus: making an ally from an enemy.

    PubMed

    Dawson, William O; Bar-Joseph, Moshe; Garnsey, Stephen M; Moreno, Pedro

    2015-01-01

    Virus diseases of perennial trees and vines have characteristics not amenable to study using small model annual plants. Unique disease symptoms such as graft incompatibilities and stem pitting cause considerable crop losses. Also, viruses in these long-living plants tend to accumulate complex populations of viruses and strains. Considerable progress has been made in understanding the biology and genetics of Citrus tristeza virus (CTV) and in developing it into a tool for crop protection and improvement. The diseases in tree and vine crops have commonalities for which CTV can be used to develop a baseline. The purpose of this review is to provide a necessary background of systems and reagents developed for CTV that can be used for continued progress in this area and to point out the value of the CTV-citrus system in answering important questions on plant-virus interactions and developing new methods for controlling plant diseases. PMID:25973695

  10. Two distinct evolutionary pathways for Citrus tristeza virus: recombination defines two gene modules and provides for increased genetic diversity in a narrow host range plant virus.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phylogenetic analysis of the full or partial genomic sequences of the Citrus tristeza virus (CTV) isolates T36, T68-1 and NS25 showed phylogenetic incongruities between sequences involved in viral RNA replication and those involved in movement and other viral functions. This incongruity was not fou...

  11. Evaluación de Anticuerpos Desarrollados Contra la Proteína Recombinante de la Cápside del Virus Tristeza de los Cítricos

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polyclonal antibodies specific for the recombinant coat protein (rCP) p25 gene of (Citrus tristeza virus = CTV), were developed for isolates MX08 and MX14 from México and B227 from India. The reactivity of rCP antibodies was evaluated using healthy and CTV infected tissue. The combination of rCP ant...

  12. A 5'-proximal region of the Citrus tristeza virus genome encoding two leader proteases is involved in virus superinfection exclusion.

    PubMed

    Atallah, Osama O; Kang, Sung-Hwan; El-Mohtar, Choaa A; Shilts, Turksen; Bergua, María; Folimonova, Svetlana Y

    2016-02-01

    Superinfection exclusion (SIE), a phenomenon in which a primary virus infection prevents a secondary infection with the same or closely related virus, has been observed with various viruses. Earlier we demonstrated that SIE by Citrus tristeza virus (CTV) requires viral p33 protein. In this work we show that p33 alone is not sufficient for virus exclusion. To define the additional viral components that are involved in this phenomenon, we engineered a hybrid virus in which a 5'-proximal region in the genome of the T36 isolate containing coding sequences for the two leader proteases L1 and L2 has been substituted with a corresponding region from the genome of a heterologous T68-1 isolate. Sequential inoculation of plants pre-infected with the CTV L1L2T68 hybrid with T36 CTV resulted in superinfection with the challenge virus, which indicated that the substitution of the L1-L2 coding region affected SIE ability of the virus. PMID:26748332

  13. Evolutionary analysis of genetic variation observed in citrus tristeza virus (CTV) after host passage.

    PubMed

    Sentandreu, V; Castro, J A; Ayllón, M A; Rubio, L; Guerri, J; González-Candelas, F; Moreno, P; Moya, A

    2006-05-01

    We have studied the genetic variability in two genes (p18 and p20) from two groups of Citrus tristeza virus (CTV) isolates. One group (isolates T385, T317, T318, and T305) was derived from a Spanish source by successive host passages while the other (isolates T388 and T390) was obtained after aphid transmission from a Japanese source. A total of 274 sequences were obtained for gene p18 and 451 for p20. In the corresponding phylogenetic trees, sequences derived from the severe isolates (T318, T305, and T388) clustered together and separately from those derived from mild or moderate isolates (T385, T317, and T390), regardless of their geographic origin. Hierarchical analyses of molecular variance showed that up to 53% of the total genetic variability in p18 and up to 87% of the variation in p20 could be explained by differences in the pathogenicity features of the isolates. Neutrality tests revealed that different selection forces had been acting between isolates and between genes, with purifying selection being suggested for p18 from isolates T385 and T390 and for p20 from isolates T385, T317, and T388, and balancing selection for p18 from isolates T318, T305, and T388 and for p20 from isolates T318 and T390. Furthermore, several models of codon selection were observed, with purifying selection being the most notable one, compatible with low effective population size of the virus populations resulting from transmission bottlenecks. We found no evidence of recombination playing a significant role during p18 and p20 evolution in these isolates. These results suggest that hosts can be an important evolutionary factor for CTV isolates. PMID:16329002

  14. Transcriptome analysis of sweet orange trees infected with ‘Candidatus Liberibacter asiaticus’ and two strains of citrus tristeza virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Huanglongbing (HLB) and tristeza, are diseases of citrus caused by a member of the a-proteobacteria, ‘Candidatus Liberibacter asiaticus’ (CaLas), and Citrus tristeza virus (CTV) respectively. HLB is a devastating disease, but CTV strains vary from very severe to very mild. Both CaLas and CTV are p...

  15. Past and future of a century old Citrus Tristeza Virus collection: A California citrus germplasm tale

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The California Citrus Clonal Protection Program (CCPP), Riverside, CA provides a mechanism for introduction and distribution of citrus germplasm from any citrus-growing area of the world to California for use in research, variety improvement, or by industry. Citrus tristeza virus (CTV) is a serious ...

  16. Lateral flow immunoassay for the rapid detection of citrus tristeza virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A lateral flow methodology was developed using gold nanoparticles for rapid detection of Citrus tristeza virus (CTV). The test strip was based on a sandwich immunoassay and could be accomplished within 10 minutes. A sample was considered negative for CTV when only the control line appeared; whereas,...

  17. Genetic diversity and evidence for recent modular recombination in Hawaiian Citrus tristeza virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Hawaiian Islands are home to a widespread and diverse population of Citrus tristeza virus (CTV), an economically important pathogen of citrus. In this study we quantified the genetic diversity of two CTV genes and determined the complete genomic sequence for two strains of Hawaiian CTV. The nucl...

  18. Etiology, background, worldwide situation and control of Citrus Tristeza virus and its vectors

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is readily graft-transmissible and, in nature, is spread by aphid vectors in a semi-persistent manner. CTV-decline has killed >85 million citrus trees grown on sour orange rootstock worldwide. Citrus in these areas must be grown on CTV-tolerant or resistant rootstocks. ...

  19. Citrus tristeza virus (CTV) Causing Proteomic and Enzymatic Changes in Sweet Orange Variety “Westin”

    PubMed Central

    Dória, Milena Santos; de Sousa, Aurizângela Oliveira; Barbosa, Cristiane de Jesus; Costa, Márcio Gilberto Cardoso; Gesteira, Abelmon da Silva; Souza, Regina Martins; Freitas, Ana Camila Oliveira; Pirovani, Carlos Priminho

    2015-01-01

    Citrus Tristeza disease, caused by CTV (Citrus tristeza virus), committs citrus plantations around the world and specifically attacks phloem tissues of the plant. The virus exists as a mixture of more or less severe variants, which may or may not cause symptoms of Tristeza. The objective of this study was to analyze the changes caused by CTV in the proteome of stems of sweet orange, as well as in the activity and gene expression of antioxidant enzymes. The CTV-infected sweet orange displayed mild symptoms, which were characterized by the presence of sparse stem pitting throughout their stems. The presence of virus was confirmed by RT-PCR. Proteomic analysis by 2DE-PAGE-MS / MS revealed the identity of 40 proteins differentially expressed between CTV- infected and -non-infected samples. Of these, 33 were up-regulated and 7 were down-regulated in CTV-infected samples. Among the proteins identified stands out a specific from the virus, the coat protein. Other proteins identified are involved with oxidative stress and for this their enzymatic activity was measured. The activity of superoxide dismutase (SOD) was higher in CTV-infected samples, as catalase (CAT) showed higher activity in uninfected samples. The activity of guaiacol peroxidase (GPX) did not vary significantly between samples. However, ascorbate peroxidase (APX) was more active in the infected samples. The relative expression of the genes encoding CAT, SOD, APX and GPX was analyzed by quantitative real time PCR (RT-qPCR). The CTV-infected samples showed greater accumulation of transcripts, except for the CAT gene. This gene showed higher expression in the uninfected samples. Taken together, it can be concluded that the CTV affects the protein profile and activity and gene expression of antioxidant enzymes in plants infected by this virus. PMID:26207751

  20. Citrus tristeza virus (CTV) Causing Proteomic and Enzymatic Changes in Sweet Orange Variety "Westin".

    PubMed

    Dória, Milena Santos; Sousa, Aurizângela Oliveira de; Barbosa, Cristiane de Jesus; Costa, Márcio Gilberto Cardoso; Gesteira, Abelmon da Silva; Souza, Regina Martins; Freitas, Ana Camila Oliveira; Pirovani, Carlos Priminho

    2015-01-01

    Citrus Tristeza disease, caused by CTV (Citrus tristeza virus), committs citrus plantations around the world and specifically attacks phloem tissues of the plant. The virus exists as a mixture of more or less severe variants, which may or may not cause symptoms of Tristeza. The objective of this study was to analyze the changes caused by CTV in the proteome of stems of sweet orange, as well as in the activity and gene expression of antioxidant enzymes. The CTV-infected sweet orange displayed mild symptoms, which were characterized by the presence of sparse stem pitting throughout their stems. The presence of virus was confirmed by RT-PCR. Proteomic analysis by 2DE-PAGE-MS / MS revealed the identity of 40 proteins differentially expressed between CTV- infected and -non-infected samples. Of these, 33 were up-regulated and 7 were down-regulated in CTV-infected samples. Among the proteins identified stands out a specific from the virus, the coat protein. Other proteins identified are involved with oxidative stress and for this their enzymatic activity was measured. The activity of superoxide dismutase (SOD) was higher in CTV-infected samples, as catalase (CAT) showed higher activity in uninfected samples. The activity of guaiacol peroxidase (GPX) did not vary significantly between samples. However, ascorbate peroxidase (APX) was more active in the infected samples. The relative expression of the genes encoding CAT, SOD, APX and GPX was analyzed by quantitative real time PCR (RT-qPCR). The CTV-infected samples showed greater accumulation of transcripts, except for the CAT gene. This gene showed higher expression in the uninfected samples. Taken together, it can be concluded that the CTV affects the protein profile and activity and gene expression of antioxidant enzymes in plants infected by this virus. PMID:26207751

  1. Differential stylet penetration behaviors of two Aphis gossypii biotypes in relation to host or vector infection with Citrus tristeza virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is one of the most important citrus disease agents worldwide. The impact of CTV on American agriculture has been significant, affecting 50 million trees with economic losses of several hundred million dollars. In California, this virus is predominantly transmitted by tw...

  2. Molecular diversity of Citrus tristeza virus (CTV) strains collected over the past 50 years and maintained in CTV collections in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tristeza, caused by Citrus tristeza virus (CTV), is a serious citrus disease worldwide. Because severe strains of CTV reduce fruit production and quality, CTV has been eliminated from citrus germplasm sources by a certification program. CTV is also a regulated pathogen in quarantine zones and infec...

  3. Exploring the limits of vector construction based on Citrus tristeza virus.

    PubMed

    El-Mohtar, Choaa; Dawson, William O

    2014-01-01

    We examined the limits of manipulation of the Citrus tristeza virus (CTV) genome for expressing foreign genes in plants. We previously created a vector with a foreign gene cassette inserted between the major and minor coat protein genes, which is position 6 from the 3' terminus. Yet, this virus has 10 3'-genes with several other potential locations for expression of foreign genes. Since genes positioned closer to the 3' terminus tend to be expressed in greater amounts, there were opportunities for producing greater amounts of foreign protein. We found that the virus tolerated insertions of an extra gene in most positions within the 3' region of the genome with substantially increased levels of gene product produced throughout citrus trees. CTV was amazingly tolerant to manipulation resulting in a suite of stable transient expression vectors, each with advantages for specific uses and sizes of foreign genes in citrus trees. PMID:24314658

  4. Investigation of seedling yellows cross protection by mild components of the Dekopon strain of Citrus tristeza virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Virulent strains of Citrus tristeza virus (CTV) can be controlled by pre-infection by mild strains of CTV which is called cross protection. However, the mode of action of cross protection is unknown and its durability unpredictable. RNA silencing is a regulatory mechanism to maintain genome integri...

  5. Studies of Seedling Yellows Amelioration of Citrus tristeza virus Strain Mixtures to Elucidate Mechanisms of Cross Protection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) cross-protection involves a mild strain of CTV preventing or interfering with infection or symptom expression by a severe strain. It is used to protect citrus when virulent stem pitting strains of CTV and efficient aphid vectors are endemic. However, the mode of action ...

  6. Fighting HLB with Citrus tristeza virus (CTV): Heterogeneity in the genome ends of CTV is an important consideration

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As California prepares for a potential showdown with Huanglongbing (HLB), contemporary disease control strategies that use low inputs, yet produce high value control, are needed to manage the disease. With biotechnology, Citrus tristeza virus (CTV) may be developed into a tool for protection or trea...

  7. Elevated Background in DAS-I ELISA for the Detection of Citrus Tristeza Virus in Mandarin Varieties

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Healthy tissue extracts from mandarin orange varieties occasionally resulted in elevated absorbance which can exceed twice that of healthy sweet orange tissue extracts in DAS-I-ELISA detection with an expressed coat protein polyclonal antiserum of Citrus tristeza virus (CTV). This problem occurs se...

  8. The conundrum of a unique protein encoded by citrus tristeza virus that is dispensable for infection of most hosts yet shows characteristics of a viral movement protein.

    PubMed

    Bak, Aurélie; Folimonova, Svetlana Y

    2015-11-01

    Citrus tristeza virus (CTV), one of the most economically important viruses, produces a unique protein, p33, which is encoded only in the genomes of isolates of CTV. Recently, we demonstrated that membrane association of the p33 protein confers virus ability to extend its host range. In this work we show that p33 shares characteristics of viral movement proteins. Upon expression in a host cell, the protein localizes to plasmodesmata and displays the ability to form extracellular tubules. Furthermore, p33 appears to traffic via the cellular secretory pathway and the actin network to plasmodesmata locations and is likely being recycled through the endocytic pathway. Finally, our study reveals that p33 colocalizes with a putative movement protein of CTV, the p6 protein. These results suggest a potential role of p33 as a noncanonical viral movement protein, which mediates virus translocation in the specific hosts. PMID:26210077

  9. Stem pitting and seedling yellows symptoms of Citrus tristeza virus infection may be determined by minor sequence variants.

    PubMed

    Cerni, Silvija; Ruscić, Jelena; Nolasco, Gustavo; Gatin, Zivko; Krajacić, Mladen; Skorić, Dijana

    2008-02-01

    The isolates of Citrus tristeza virus (CTV), the most destructive viral pathogen of citrus, display a high level of variability. As a result of genetic bottleneck induced by the bud-inoculation of CTV-infected material, inoculated seedlings of Citrus wilsonii Tanaka displayed different symptoms. All successfully grafted plants showed severe symptoms of stem pitting and seedling yellows, while plants in which inoculated buds died displayed mild symptoms. Since complex CTV population structure was detected in the parental host, the aim of this work was to investigate how it changed after the virus transmission, and to correlate it with observed symptoms. The coat protein gene sequence of the predominant genotype was identical in parental and grafted plants and clustered to the phylogenetic group 5 encompassing severe reference isolates. In seedlings displaying severe symptoms, the low-frequency variants clustering to other phylogenetic groups were detected, as well. Indicator plants were inoculated with buds taken from unsuccessfully grafted C. wilsonii seedlings. Surprisingly, they displayed no severe symptoms despite the presence of phylogenetic group 5 genomic variants. The results suggest that the appearance of severe symptoms in this case is probably induced by a complex CTV population structure found in seedlings displaying severe symptoms, and not directly by the predominant genomic variant. PMID:18074213

  10. Virus-viroid interactions: Citrus Tristeza Virus enhances the accumulation of Citrus Dwarfing Viroid in Mexican lime via virus-encoded silencing suppressors.

    PubMed

    Serra, Pedro; Bani Hashemian, Seyed M; Fagoaga, Carmen; Romero, Juan; Ruiz-Ruiz, Susana; Gorris, Maria T; Bertolini, Edson; Duran-Vila, Núria

    2014-01-01

    An assay to identify interactions between Citrus Dwarfing Viroid (CDVd) and Citrus Tristeza Virus (CTV) showed that viroid titer was enhanced by the coinfecting CTV in Mexican lime but not in etrog citron. Since CTV encodes three RNA silencing suppressors (RSSs), p23, p20 and p25, an assay using transgenic Mexican limes expressing each RSS revealed that p23 and, to a lesser extent, p25 recapitulated the effect observed with coinfections of CTV and CDVd. PMID:24227850

  11. Citrus Tristeza Virus: Survival at the Edge of the Movement Continuum▿

    PubMed Central

    Folimonova, Svetlana Y.; Folimonov, Alexey S.; Tatineni, Satyanarayana; Dawson, William O.

    2008-01-01

    Systemic invasion of plants by viruses is thought to involve two processes: cell-to-cell movement between adjacent cells and long-distance movement that allows the virus to rapidly move through sieve elements and unload at the growing parts of the plant. There is a continuum of proportions of these processes that determines the degrees of systemic infection of different plants by different viruses. We examined the systemic distribution of Citrus tristeza virus (CTV) in citrus species with a range of susceptibilities. By using a “pure” culture of CTV from a cDNA clone and green fluorescent protein-labeled virus we show that both cell-to-cell and long-distance movement are unusually limited, and the degree of limitation varies depending on the citrus host. In the more-susceptible hosts CTV infected only a small portion of phloem-associated cells, and moreover, the number of infection sites in less-susceptible citrus species was substantially decreased further, indicating that long-distance movement was reduced in those hosts. Analysis of infection foci in the two most differential citrus species, Citrus macrophylla and sour orange, revealed that in the more-susceptible host the infection foci were composed of a cluster of multiple cells, while in the less-susceptible host infection foci were usually single cells, suggesting that essentially no cell-to-cell movement occurred in the latter host. Thus, CTV in sour orange represents a pattern of systemic infection in which the virus appears to function with only the long-distance movement mechanism, yet is able to survive in nature. PMID:18434397

  12. Calculation of diagnostic parameters of advanced serological and molecular tissue-print methods for detection of Citrus tristeza virus. A model for other plant pathogens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is one of the most important virus diseases which affect citrus. Control of CTV in Spain and central California is achieved by planting virus-free citrus on CTV-tolerant or -resistant rootstocks. Quarantine and certification programs remain essential to avoid importation ...

  13. Detection of Citrus tristeza virus by using fluorescence resonance energy transfer-based biosensor.

    PubMed

    Shojaei, Taha Roodbar; Salleh, Mohamad Amran Mohd; Sijam, Kamaruzaman; Rahim, Raha Abdul; Mohsenifar, Afshin; Safarnejad, Reza; Tabatabaei, Meisam

    2016-12-01

    Due to the low titer or uneven distribution of Citrus tristeza virus (CTV) in field samples, detection of CTV by using conventional detection techniques may be difficult. Therefore, in the present work, the cadmium-telluride quantum dots (QDs) was conjugated with a specific antibody against coat protein (CP) of CTV, and the CP were immobilized on the surface of gold nanoparticles (AuNPs) to develop a specific and sensitive fluorescence resonance energy transfer (FRET)-based nanobiosensor for detecting CTV. The maximum FRET efficiency for the developed nano-biosensor was observed at 60% in AuNPs-CP/QDs-Ab ratio of 1:8.5. The designed system showed higher sensitivity and specificity over enzyme linked immunosorbent assay (ELISA) with a limit of detection of 0.13μgmL(-1) and 93% and 94% sensitivity and specificity, respectively. As designed sensor is rapid, sensitive, specific and efficient in detecting CTV, this could be envisioned for diagnostic applications, surveillance and plant certification program. PMID:27380305

  14. Rapid Differentiation and Identification of Potential Severe Strains of Citrus tristeza Virus by Real-Time Reverse Transcription Polymerase Chain Reaction Assays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A multiplex Taqman®-based real-time reverse transcription (RT) polymerase chain reaction (PCR) assay was developed to detect all strains of Citrus tristeza virus (CTV) and to identify potentially severe strains of the virus. A CTV TaqMan probe (CTV-CY5) based on the coat protein (CP) gene sequences...

  15. The Pathogenicity Determinant of Citrus Tristeza Virus Causing the Seedling Yellows Syndrome is Located at the 3’-Terminal Region of the Viral Genome

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) (genus Closterovirus, family Closteroviridae) causes some of the more important viral diseases of citrus worldwide. The ability to map disease-inducing determinants of CTV is needed to develop better diagnostic and disease control procedures. A distinctive phenotype of s...

  16. Heterologous Minor Coat Proteins of Citrus Tristeza Virus Strains Affect Encapsidation, but the Coexpression of HSP70h and p61 Restores Encapsidation to Wild-Type Levels

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The long flexuous bipolar virions of Citrus tristeza virus (CTV), a Closterovirus, are encapsidated with two capsid proteins at opposite ends: the minor coat protein (CPm) encapsidates the 5’ 630 nts of the genomic RNA and the major coat protein encapsidates the remainder of the genome. In this stud...

  17. Quantitative Detection of Citrus tristeza virus (CTV) in Citrus and Aphids by Real-time Reverse Transcription-PCR (TaqMan®)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Routine detection of Citrus tristeza virus (CTV) is by enzyme-linked immunosorbent (ELISA) and direct tissue blot immunoassays. Reverse transcription (RT) polymerase chain reaction (PCR) has also been developed for CTV detection which is more sensitive than serology. We developed a quantitative an...

  18. Genetic diversity of citrus tristeza virus from cross-protected and unprotected citrus trees after 20 years of natural challenge in Peru

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Quick decline, caused by Citrus tristeza virus (CTV), devastated Peruvian citrus on sour orange rootstock between 1950 and 1965. A second epidemic occurred between 1965 to 1985 due to spread of CTV strains causing severe stem pitting (SP) on branches and trunks, regardless of rootstock. SP weakens t...

  19. Comparison of gene expression changes in susceptible, tolerant, and resistant hosts in response to infection with citrus tristeza virus and huanglongbing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The pathogens Candidatus Liberibacter asiaticus (Las) and Citrus tristeza virus (CTV) are both phloem limited and have significant economic impact on citrus production wherever they are found. Studies of host resistance have indicated that Poncirus trifoliata has tolerance or resistance to both path...

  20. Profiling of the small RNA populations derived from sour orange seedlings cross-protected against seedling yellows strains of Citrus tristeza virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Control of Citrus tristeza virus (CTV) in central California changed in 2009 from removal of all CTV-infected trees to only those which react positive in tests with selective probes for potentially severe CTV strains. Therefore, new strategies for CTV control are needed. Greenhouse tests have show...

  1. Accumulation of a 5’ proximal subgenomic RNA of Citrus tristeza virus is correlated with encapsidation by the minor coat protein

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During replication, Citrus tristeza virus (CTV) produces large amounts of two unusual subgenomic (sg) RNAs that are positive-stranded and 5' -coterminal. Although these RNAs are produced in similar amounts and are similar in size, with LMT1 (~750 nt) only slightly larger than LMT2 (~650), we found ...

  2. Strains of Citrus tristeza virus do not exclude superinfection by other strains of the virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Superinfection exclusion or homologous interference, a phenomenon in which a primary viral infection prevents a secondary infection with the same or closely-related virus, has been observed commonly for viruses in various systems, including viruses of bacteria, plants, and animals. With plant viruse...

  3. Rapid differentiation and identification of potential severe strains of Citrus tristeza virus by real-time reverse transcription-polymerase chain reaction assays.

    PubMed

    Yokomi, R K; Saponari, M; Sieburth, P J

    2010-04-01

    A multiplex Taqman-based real-time reverse transcription (RT) polymerase chain reaction (PCR) assay was developed to identify potential severe strains of Citrus tristeza virus (CTV) and separate genotypes that react with the monoclonal antibody MCA13. Three strain-specific probes were developed using intergene sequences between the major and minor coat protein genes (CPi) in a multiplex reaction. Probe CPi-VT3 was designed for VT and T3 genotypes; probe CPi-T36 for T36 genotypes; and probe CPi-T36-NS to identify isolates in an outgroup clade of T36-like genotypes mild in California. Total nucleic acids extracted by chromatography on silica particles, sodium dodecyl sulfate-potassium acetate, and CTV virion immunocapture all yielded high quality templates for real-time PCR detection of CTV. These assays successfully differentiated CTV isolates from California, Florida, and a large panel of CTV isolates from an international collection maintained in Beltsville, MD. The utility of the assay was validated using field isolates collected in California and Florida. PMID:20205535

  4. Synthesis and Assessment of DNA/Silver Nanoclusters Probes for Optimal and Selective Detection of Tristeza Virus Mild Strains.

    PubMed

    Shokri, Ehsan; Hosseini, Morteza; Faridbod, Farnoush; Rahaie, Mahdi

    2016-09-01

    Citrus Tristeza virus (CTV) is one of the most destructive pathogens worldwide that exist as a mixture of malicious (Sever) and tolerable (Mild) strains. Mild strains of CTV can be used to immunize healthy plants from more Severe strains damage. Recently, innovative methods based on the fluorescent properties of DNA/silver nanoclusters have been developed for molecular detection purposes. In this study, a simple procedure was followed to create more active DNA/AgNCs probe for accurate and selective detection of Tristeza Mild-RNA. To this end, four distinct DNA emitter scaffolds (C12, Red, Green, Yellow) were tethered to the Mild capture sequence and investigated in various buffers in order to find highly emissive combinations. Then, to achieve specific and reliable results, several chemical additives, including organic solvents, PEG and organo-soluble salts were used to enhance control fluorescence signals and optimize the hybridization solution. The data showed that, under adjusted conditions, the target sensitivity is enhanced by a factor of five and the high discrimination between Mild and Severe RNAs were obtained. The emission ratio of the DNA/AgNCs was dropped in the presence of target RNAs and I0/I intensity linearly ranged from 1.5 × 10(-8) M to 1.8 × 10(-6) M with the detection limit of 4.3 × 10(-9) M. PMID:27349801

  5. Development and validation of a multiplex reverse transcription quantitative PCR (RT-qPCR) assay for the rapid detection of Citrus tristeza virus, Citrus psorosis virus, and Citrus leaf blotch virus.

    PubMed

    Osman, Fatima; Hodzic, Emir; Kwon, Sun-Jung; Wang, Jinbo; Vidalakis, Georgios

    2015-08-01

    A single real-time multiplex reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay for the simultaneous detection of Citrus tristeza virus (CTV), Citrus psorosis virus (CPsV), and Citrus leaf blotch virus (CLBV) was developed and validated using three different fluorescently labeled minor groove binding qPCR probes. To increase the detection reliability, coat protein (CP) genes from large number of different isolates of CTV, CPsV and CLBV were sequenced and a multiple sequence alignment was generated with corresponding CP sequences from the GenBank and a robust multiplex RT-qPCR assay was designed. The capacity of the multiplex RT-qPCR assay in detecting the viruses was compared to singleplex RT-qPCR designed specifically for each virus and was assessed using multiple virus isolates from diverse geographical regions and citrus species as well as graft-inoculated citrus plants infected with various combination of the three viruses. No significant difference in detection limits was found and specificity was not affected by the inclusion of the three assays in a multiplex RT-qPCR reaction. Comparison of the viral load for each virus using singleplex and multiplex RT-qPCR assays, revealed no significant differences between the two assays in virus detection. No significant difference in Cq values was detected when using one-step and two-step multiplex RT-qPCR detection formats. Optimizing the RNA extraction technique for citrus tissues and testing the quality of the extracted RNA using RT-qPCR targeting the cytochrome oxidase citrus gene as an RNA specific internal control proved to generate better diagnostic assays. Results showed that the developed multiplex RT-qPCR can streamline viruses testing of citrus nursery stock by replacing three separate singleplex assays, thus reducing time and labor while retaining the same sensitivity and specificity. The three targeted RNA viruses are regulated pathogens for California's mandatory "Section 3701

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

    PubMed Central

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

    2013-01-01

    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

  7. Accumulation of a 5' proximal subgenomic RNA of Citrus tristeza virus is correlated with encapsidation by the minor coat protein.

    PubMed

    Gowda, Siddarame; Tatineni, Satyanarayana; Folimonova, Svetlana Y; Hilf, Mark E; Dawson, William O

    2009-06-20

    During replication, Citrus tristeza virus (CTV) produces large amounts of two unusual subgenomic (sg) RNAs that are positive-stranded and 5' coterminal. Although these RNAs are produced in similar amounts and are similar in size, with LMT1 ( approximately 750 nt) only slightly larger than LMT2 ( approximately 650), we found that the similar sgRNAs are produced differently. We previously showed that the LMT1 RNA is produced by premature termination during genomic RNA synthesis. However, LMT2 production was found to correlate with virion assembly instead of RNA replication. The time course of accumulation of the LMT2 RNA occurred late, coinciding with virion accumulation. The long flexuous virions of CTV contain two coat proteins that encapsidate the virions in a polar manner. The major coat protein encapsidates approximately 97% of the virion, while the minor capsid protein encapsidates the remainder of the genome beginning in the 5' non-translated region with the transition zone at approximately 630 nucleotides from the 5' end. The section of the virion RNA that was encapsidated by CPm was identical in size to the LMT2 RNA, suggesting that the LMT2 RNA represented a portion of the viral RNA protected by CPm encapsidation. Mutations that abrogated encapsidation by CPm also abolished the accumulation of LMT2 RNA. Thus, these two unusual but similar RNAs are produced via different pathways, one from RNA replication and one processed by the virion assembly process. To our knowledge, this represents the first evidence of a viral RNA processed by the assembly mechanism. PMID:19446304

  8. Symptoms induced by transgenic expression of p23 from Citrus tristeza virus in phloem-associated cells of Mexican lime mimic virus infection without the aberrations accompanying constitutive expression.

    PubMed

    Soler, Nuria; Fagoaga, Carmen; López, Carmelo; Moreno, Pedro; Navarro, Luis; Flores, Ricardo; Peña, Leandro

    2015-05-01

    Citrus tristeza virus (CTV) is phloem restricted in natural citrus hosts. The 23-kDa protein (p23) encoded by the virus is an RNA silencing suppressor and a pathogenicity determinant. The expression of p23, or its N-terminal 157-amino-acid fragment comprising the zinc finger and flanking basic motifs, driven by the constitutive 35S promoter of cauliflower mosaic virus, induces CTV-like symptoms and other aberrations in transgenic citrus. To better define the role of p23 in CTV pathogenesis, we compared the phenotypes of Mexican lime transformed with p23-derived transgenes from the severe T36 and mild T317 CTV isolates under the control of the phloem-specific promoter from Commelina yellow mottle virus (CoYMV) or the 35S promoter. Expression of the constructs restricted to the phloem induced a phenotype resembling CTV-specific symptoms (vein clearing and necrosis, and stem pitting), but not the non-specific aberrations (such as mature leaf epinasty and yellow pinpoints, growth cessation and apical necrosis) observed when p23 was ectopically expressed. Furthermore, vein necrosis and stem pitting in Mexican lime appeared to be specifically associated with p23 from T36. Phloem-specific accumulation of the p23Δ158-209(T36) fragment was sufficient to induce the same anomalies, indicating that the region comprising the N-terminal 157 amino acids of p23 is responsible (at least in part) for the vein clearing, stem pitting and, possibly, vein corking in this host. PMID:25171669

  9. Volatile Organic Compound (VOC) profiling of Citrus tristeza virus (CTV) infection in sweet orange citrus varietals using thermal desorption gas chromatography time of flight mass spectrometry (TD-GC/TOF-MS)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is a plant pathogen which predominately infects economically important citrus crops such as sweet orange, clementine, lime and grapefruit varietals. Within the last 70 years, an estimated 100 million citrus trees on sour orange rootstock have been destroyed due to CTV inf...

  10. Booster immunization with a partially purified citrus tristeza virus (CTV) preparation after priming with recombinant CTV coat protein enhances the binding capacity of capture antibodies by ELISA.

    PubMed

    Bar-Joseph, M; Filatov, V; Gofman, R; Guang, Y; Hadjinicolis, A; Mawassi, M; Gootwine, E; Weisman, Y; Malkinson, M

    1997-08-01

    Groups of rabbits and young lambs were immunized subcutaneously and intramuscularly with a recombinant citrus tristeza virus (CTV) coat protein (rCTV-CP) antigen. Three weeks after primary immunization the animals were divided into two groups that were boosted either with rCTV-CP or with a partially purified preparation of CTV particles (ppCTV). Twelve and 15 days after the last injection, the animals were bled and the binding capacity of the antisera for CTV detection was examined for capture antibodies by the indirect ELISA. Considerably higher ELISA titers were obtained from animals that were boosted with ppCTV than with rCP. Boosting with partially purified native antigens after priming with recombinant antigens is expected to extend the applicability of the antisera for detecting other structural and non-structural viral antigens by trapping ELISA. PMID:9274814

  11. 2006 University Citrus Pest Management Guide: Tristeza

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) is a major cause of the decline and eventual death of trees on sour orange rootstocks. Initially, affected trees have small leaves and twig dieback. Diseased trees often produce a crop of very small fruit. Eventually, large limbs die back and the tree gradually declines...

  12. Sequence Analysis of a 282-Kilobase Region Surrounding the Citrus Tristeza Virus Resistance Gene (Ctv) Locus in Poncirus trifoliata L. Raf.1

    PubMed Central

    Yang, Zhong-Nan; Ye, Xin-Rong; Molina, Joe; Roose, Mikeal L.; Mirkov, T. Erik

    2003-01-01

    Citrus tristeza virus (CTV) is the major virus pathogen causing significant economic damage to citrus worldwide, and a single dominant gene, Ctv, provides broad spectrum resistance to CTV in Poncirus trifoliata L. Raf. Ctv was physically mapped to a 282-kb region using a P. trifoliata bacterial artificial chromosome library. This region was completely sequenced to about 8× coverage using a shotgun sequencing strategy and primer walking for gap closure. Sequence analysis predicts 22 putative genes, two mutator-like transposons and eight retrotransposons. This sequence analysis also revealed some interesting features of this region of the P. trifoliata genome: a disease resistance gene cluster with seven members and eight retrotransposons clustered in a 125-kb gene-poor region. Comparative sequence analysis suggests that six genes in the Ctv region have significant sequence similarity with their orthologs in bacterial artificial chromosome clones F7H2 and F21T11 from Arabidopsis chromosome I. However, the analysis of gene colinearity between P. trifoliata and Arabidopsis indicates that Arabidopsis genome sequence information may be of limited use for positional gene cloning in P. trifoliata and citrus. Analysis of candidate genes for Ctv is also discussed. PMID:12586873

  13. Sequence analysis of a 282-kilobase region surrounding the citrus Tristeza virus resistance gene (Ctv) locus in Poncirus trifoliata L. Raf.

    PubMed

    Yang, Zhong-Nan; Ye, Xin-Rong; Molina, Joe; Roose, Mikeal L; Mirkov, T Erik

    2003-02-01

    Citrus tristeza virus (CTV) is the major virus pathogen causing significant economic damage to citrus worldwide, and a single dominant gene, Ctv, provides broad spectrum resistance to CTV in Poncirus trifoliata L. Raf. Ctv was physically mapped to a 282-kb region using a P. trifoliata bacterial artificial chromosome library. This region was completely sequenced to about 8x coverage using a shotgun sequencing strategy and primer walking for gap closure. Sequence analysis predicts 22 putative genes, two mutator-like transposons and eight retrotransposons. This sequence analysis also revealed some interesting features of this region of the P. trifoliata genome: a disease resistance gene cluster with seven members and eight retrotransposons clustered in a 125-kb gene-poor region. Comparative sequence analysis suggests that six genes in the Ctv region have significant sequence similarity with their orthologs in bacterial artificial chromosome clones F7H2 and F21T11 from Arabidopsis chromosome I. However, the analysis of gene colinearity between P. trifoliata and Arabidopsis indicates that Arabidopsis genome sequence information may be of limited use for positional gene cloning in P. trifoliata and citrus. Analysis of candidate genes for Ctv is also discussed. PMID:12586873

  14. 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

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

    2014-04-20

    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

  15. Chlorella viruses isolated in China

    SciTech Connect

    Zhang, Y.; Burbank, D.E.; Van Etten, J.L. )

    1988-09-01

    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.

  16. Heterologous minor coat proteins of Citrus tristeza virus strains affect encapsidation, but the coexpression of HSP70h and p61 restores encapsidation to wild-type levels.

    PubMed

    Tatineni, Satyanarayana; Gowda, Siddarame; Dawson, William O

    2010-07-01

    The long flexuous bipolar virions of Citrus tristeza virus (CTV), a Closterovirus, are encapsidated with two capsid proteins at opposite ends: the minor coat protein (CPm) encapsidates the 5' 630 nts of the genomic RNA and the major coat protein encapsidates the remainder of the genome. In this study, we found encapsidation of CTV CPm in the absence of other assembly-related proteins is highly specific in contrast to most plant viruses that allow virion assembly by a range of heterologous coat proteins. Heterologous CPms with 95-96% amino acid identity from related strains in CTV-CPm, a replicon with CPm as the only assembly-related ORF, either failed to initiate encapsidation or reduced encapsidation substantially. Substitution of subsets of amino acids revealed that the amino acids that differ between positions 121 and 180 of the VT strain, and 61 and 120 of the T3 strain were involved in specific encapsidation. We further mapped the specific encapsidation to a single amino acid: mutation of methionine(165) to threonine (VT type) or serine(105) to proline (T3 type) in CTV-CPm failed to form nucleocapsids. However, the heterologous CPm in combination with both HSP70h and p61 proteins, but not HSP70h or p61 alone, encapsidated at wild-type levels, suggesting that specific encapsidation by CPm was mitigated by the combination of HSP70h and p61. Thus, in addition to the previously described functions of HSP70h and p61 of greatly enhanced virion formation and restriction of CPm encapsidation to the 5' 630 nts of the genomic RNA, these proteins facilitate encapsidation by heterologous CPms. PMID:20399478

  17. Virus diffusion in isolation rooms.

    PubMed

    Kao, P H; Yang, R J

    2006-03-01

    In hospitals, the ventilation of isolation rooms operating under closed-door conditions is vital if the spread of viruses and infection is to be contained. Engineering simulation, which employs computational fluid dynamics, provides a convenient means of investigating airflow behaviour in isolation rooms for various ventilation arrangements. A cough model was constructed to permit the numerical simulation of virus diffusion inside an isolation room for different ventilation system configurations. An analysis of the region of droplet fallout and the dilution time of virus diffusion of coughed gas in the isolation room was also performed for each ventilation arrangement. The numerical results presented in this paper indicate that the parallel-directional airflow pattern is the most effective means of controlling flows containing virus droplets. Additionally, staggering the positions of the supply vents at the door end of the room relative to the exhaust vents on the wall behind the bed head provides effective infection control and containment. These results suggest that this particular ventilation arrangement enhances the safety of staff when performing medical treatments within isolation rooms. PMID:16359753

  18. Characterization of Recently Introduced HLB and CTV Isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Severe stem-pitting isolates of Citrus tristeza virus (CTV) were found in Florida citrus 5 years ago, followed by the discovery 2 years later of the citrus greening disease (huanglongbing: HLB) caused by the bacterium Candidatus Liberibacter asiaticus. The new CTV isolates are members of the VT grou...

  19. Recombination defines two gene modules and provides for increased genetic diversity in a narrow host range plant virus.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Citrus tristeza virus (CTV) isolates T36, T68-1 and NS25 showed phylogenetic incongruities between sequences involved in viral RNA replication and those involved in movement and other viral functions. This incongruity was not found in the sequences of isolates T3, T30, T385, VT and T318A. Distance...

  20. Virus isolation and propagation in embryonating eggs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The embryonating egg is one of the most versatile, easy to work with, and widely used host systems for the isolation and propagation of avian viruses. The embryonating chicken egg (ECE) is the most commonly available system that is both specific pathogen free and supports the replication of viruses...

  1. Refinement of the Citrus tristeza virus resistance gene (Ctv) positional map in Poncirus trifoliata and generation of transgenic grapefruit (Citrus paradisi) plant lines with candidate resistance genes in this region.

    PubMed

    Rai, Mamta

    2006-06-01

    Citrus tristeza virus (CTV) is a major pathogen of Citrus. A single dominant gene Ctv present in the trifoliate relative of Citrus, Poncirus trifoliata confers broad spectrum resistance against CTV. Refinement of genetic maps has delimited this gene to a 121 kb region, comprising of ten candidate Ctv resistance genes. The ten candidate genes were individually cloned in Agrobacterium based binary vector and transformed into three CTV susceptible grapefruit varieties. Two of the candidate R-genes, R-2 and R-3 are exclusively expressed in transgenic plants and in Poncirus trifoliata, while five other genes are also expressed in non-transformed Citrus controls. Northern blotting with a CTV derived probe for assessment of infection in virus inoculated plants over a span of three growth periods, each comprising of six to eight weeks, indicates either an absence of initiation of infection or it's slow spread in R-2 plant lines or an initial appearance of infection and it's subsequent obliteration in some R-1 and R-4 plant lines. Limited genome walk up- and downstream form R-1 gene, based on it's 100% sequence identity between Poncirus and Citrus, indicates promoter identity of 92% between the two varieties. Further upstream and downstream sequencing indicates the presence of an O-methyl transferase and a Copia like gene respectively in Citrus instead of the amino acid transporter like gene upstream and a sugar transporter like gene downstream in Poncirus. The possibility of recombinations in the resistance locus of Citrus and the need for consistent monitoring for virus infection and gene expression in the transgenic Citrus trees is discussed. PMID:16830176

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

    PubMed Central

    Lee, Richard F.; Keremane, Manjunath L.

    2013-01-01

    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

  3. Citrus tristeza virus infection induces the accumulation of viral small RNAs (21-24-nt) mapping preferentially at the 3'-terminal region of the genomic RNA and affects the host small RNA profile.

    PubMed

    Ruiz-Ruiz, Susana; Navarro, Beatriz; Gisel, Andreas; Peña, Leandro; Navarro, Luis; Moreno, Pedro; Di Serio, Francesco; Flores, Ricardo

    2011-04-01

    To get an insight into the host RNA silencing defense induced by Citrus tristeza virus (CTV) and into the counter defensive reaction mediated by its three silencing suppressors (p25, p20 and p23), we have examined by deep sequencing (Solexa-Illumina) the small RNAs (sRNAs) in three virus-host combinations. Our data show that CTV sRNAs: (i) represent more than 50% of the total sRNAs in Mexican lime and sweet orange (where CTV reaches relatively high titers), but only 3.5% in sour orange (where the CTV titer is significantly lower), (ii) are predominantly of 21-22-nt, with a biased distribution of their 5' nucleotide and with those of (+) polarity accumulating in a moderate excess, and (iii) derive from essentially all the CTV genome (ca. 20 kb), as revealed by its complete reconstruction from viral sRNA contigs, but adopt an asymmetric distribution with a prominent hotspot covering approximately the 3'-terminal 2,500 nt. These results suggest that the citrus homologues of Dicer-like (DCL) 4 and 2 most likely mediate the genesis of the 21 and 22 nt CTV sRNAs, respectively, and show that both ribonucleases act not only on the genomic RNA but also on the 3' co-terminal subgenomic RNAs and, particularly, on their double-stranded forms. The plant sRNA profile, very similar and dominated by the 24-nt sRNAs in the three mock-inoculated controls, was minimally affected by CTV infection in sour orange, but exhibited a significant reduction of the 24-nt sRNAs in Mexican lime and sweet orange. We have also identified novel citrus miRNAs and determined how CTV influences their accumulation. PMID:21327514

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

    PubMed Central

    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

    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

  5. Sequence diversity of wheat mosaic virus isolates.

    PubMed

    Stewart, Lucy R

    2016-02-01

    Wheat mosaic virus (WMoV), transmitted by eriophyid wheat curl mites (Aceria tosichella) is the causal agent of High Plains disease in wheat and maize. WMoV and other members of the genus Emaravirus evaded thorough molecular characterization for many years due to the experimental challenges of mite transmission and manipulating multisegmented negative sense RNA genomes. Recently, the complete genome sequence of a Nebraska isolate of WMoV revealed eight segments, plus a variant sequence of the nucleocapsid protein-encoding segment. Here, near-complete and partial consensus sequences of five more WMoV isolates are reported and compared to the Nebraska isolate: an Ohio maize isolate (GG1), a Kansas barley isolate (KS7), and three Ohio wheat isolates (H1, K1, W1). Results show two distinct groups of WMoV isolates: Ohio wheat isolate RNA segments had 84% or lower nucleotide sequence identity to the NE isolate, whereas GG1 and KS7 had 98% or higher nucleotide sequence identity to the NE isolate. Knowledge of the sequence variability of WMoV isolates is a step toward understanding virus biology, and potentially explaining observed biological variation. PMID:26590326

  6. Isolation of ancestral sylvatic dengue virus type 1, Malaysia.

    PubMed

    Teoh, Boon-Teong; Sam, Sing-Sin; Abd-Jamil, Juraina; AbuBakar, Sazaly

    2010-11-01

    Ancestral sylvatic dengue virus type 1, which was isolated from a monkey in 1972, was isolated from a patient with dengue fever in Malaysia. The virus is neutralized by serum of patients with endemic DENV-1 infection. Rare isolation of this virus suggests a limited spillover infection from an otherwise restricted sylvatic cycle. PMID:21029545

  7. Molecular characterization of Korean rabies virus isolates

    PubMed Central

    Park, Young-Nam; Hong, Gyeong-Soo; Kang, Hee-Kyung; Oh, Yoon-I; Cho, Soo-Dong; Song, Jae-Young

    2011-01-01

    The nucleoprotein (N) and glycoprotein (G) of 11 Korean rabies virus (RABV) isolates collected from animals diagnosed with rabies between 2008 and 2009 were subjected to molecular and phylogenetic analyses. Six isolates originated from domestic animals (cattle and dogs) and five were obtained from wild free-ranging raccoon dogs. The similarities in the nucleotide sequences of the N gene among all Korean isolates ranged from 98.1 to 99.8%, while those of the G gene ranged from 97.9 to 99.3%. Based on the nucleotide analysis of the N and G genes, the Korean RABV isolates were confirmed as genotype I of Lyssavirus and classified into four distinct subgroups with high similarity. Phylogenetic analysis showed that the Korean isolates were most closely related to the non-Korean NeiMeng1025B and 857r strains, which were isolated from rabid raccoon dogs in Eastern China and Russia, respectively. These findings suggest that the Korean RABV isolates originated from a rabid raccoon dog in Northeastern Asia. Genetic analysis of the Korean RABV isolates revealed no substitutions at several antigenic sites, indicating that the isolates circulating in Korea may be pathogenic in several hosts. PMID:21368564

  8. Characterization of rabies virus isolates in Bolivia.

    PubMed

    Favi, Myriam; Nina, Aleida; Yung, Verónica; Fernández, Jorge

    2003-11-01

    In Latin America, rabies is still an important public health problem. Canine rabies, and wild animal rabies, especially transmitted by hematofagous and insectivorous bats, has become an emerging problem in the countries of this region. We received 363 samples with a laboratory-confirmed rabies diagnosis from Bolivia during l997-2001. From these, we could obtain 222 rabies virus isolates by intra-cerebral inoculation in mice. By antigenic characterization we could identify 147 isolates as variant 1, 2 isolates as variant 2, 3 isolates as variant 3, and 1 isolate as variant 5. Phylogenetic analysis of 84 isolates established that they segregated in 3 different branches, corresponding to 3 genetic variants, 78 isolates corresponding to antigenic variant 1 segregated in the same lineage as the antigenic variant 5, 2 isolates corresponding to antigenic variant 2 segregated in another lineage, and 3 isolates from antigenic variant 3 segregated in a different lineage.The genetic variant that mainly circulates in Bolivia is maintained in a cycle whose main reservoir are dogs, but it is not possible to discard the presence of other cycles, in which different species of bats or other wild mammals could be participating. PMID:14602205

  9. Analysis of Iranian Potato virus S isolates.

    PubMed

    Salari, Khadijeh; Massumi, Hossein; Heydarnejad, Jahangir; Hosseini Pour, Akbar; Varsani, Arvind

    2011-10-01

    Two hundred forty potato samples with one or more symptoms of leaf mosaic, distortion, mottling and yellowing were collected between 2005 and 2008 from seven Iranian provinces. Forty-four of these samples tested positive with double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA) using a Potato virus S (PVS) polyclonal antibody. Of these 12 isolates of PVS were selected based on the geographical location for biological and molecular characterization. The full coat protein (CP) and 11K genes from 12 PVS isolates were PCR amplified, cloned and sequenced. All 12 PVS isolates showed mosaic symptoms on Nicotiana debneyii and N. tabacum cv. Whiteburly and local lesion on Chenopodium amaranticolor, C. quinoa and C. album. The Iranian isolates share between 93 and 100% pairwise nucleotide identity with other PVS(O) isolates. Based on maximum likelihood phylogenetic analysis coupled with pairwise identity analysis, we propose 15 genotypes for the PVS(O) strain and 3 genotypes for the PVS(A) strain. PMID:21567245

  10. Genetic Transformation of Citrus Paradisi with Antisense and untranslatable RNA-dependent RNA Polymerase Genes of Citrus Tristeza Closterovirus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Expression of the RNA-dependent RNA polymerase (RdRp) of Citrus tristeza virus (CTV) was studied in vivo and in vitro using a polyclonal antiserum raised against the recombinant CTV-RdRp protein. Although 56 kDa CTV-RdRp is thought to be expressed by a +1 translational frameshift at the carboxyl te...

  11. Triticum mosaic virus isolates in the southern Great Plains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In 2006, a Wheat streak mosaic virus (WSMV)-resistant wheat variety RonL was found to have mosaic symptoms similar to WSMV. The virus inducing the symptoms was determined to be previously unknown and given the name Triticum mosaic virus (TriMV). Since, TriMV has been found in plant samples isolate...

  12. Variants of Triticum mosaic virus isolated from wheat in Colorado

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Triticum mosaic virus (TriMV) is a recently discovered virus infecting wheat. We compared the Colorado isolates C10-492 and C11-775 with the 06-123 isolate of TriMV from Kansas (TriMV-K). Comparisons were made using enzyme-linked immunosorbent assay (ELISA), infectivity assay, host range, dry weig...

  13. Status of tobacco viruses in Serbia and molecular characterization of tomato spotted wilt virus isolates.

    PubMed

    Stanković, I; Bulajić, A; Vučurović, A; Ristić, D; Milojević, K; Berenji, J; Krstić, B

    2011-01-01

    In a four-year survey to determine the presence and distribution of viruses in tobacco crops at 17 localities of the Vojvodina Province and Central Serbia, 380 samples were collected and analyzed by DAS-ELISA. Out of the seven viruses tested, tomato spotted wilt virus (TSWV), potato virus Y (PVY), tobacco mosaic virus (TMV), cucumber mosaic virus (CMV), and alfalfa mosaic virus (AMV) were detected in 37.9, 33.4, 28.7, 23.9, and 15.5% of the total tested samples, respectively. TSWV was the most frequently found virus at the localities of Central Serbia, while PVY and CMV were the most frequent viruses in the Vojvodina Province. Single infections were prevalent in years 2005-2007 and the most frequent were those of PVY. A triple combination of those viruses was most frequent mixed infection type in 2008. The presence of all five detected viruses was confirmed in selected ELISA-positive samples by RT-PCR and sequencing. The comparisons of obtained virus isolate sequences with those available in NCBI, confirmed the authenticity of serologically detected viruses. Phylogenetic analysis based on partial nucleocapsid gene sequences revealed a joint clustering of Serbian, Bulgarian and Montenegrin TSWV isolates into one geographic subpopulation, which was distinct from the other subpopulation of TSWV isolates from the rest of the European countries. The high incidence of viruses in Serbian tobacco crops highlights the importance of enhancing farmers knowledge towards better implementation of control strategies for preventing serious losses. PMID:22149499

  14. High sequence conservation among cucumber mosaic virus isolates from lily.

    PubMed

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

    2001-08-01

    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

  15. West Nile Virus Isolation in Human and Mosquitoes, Mexico

    PubMed Central

    Elizondo-Quiroga, Darwin; Davis, C. Todd; Fernandez-Salas, Ildefonso; Escobar-Lopez, Roman; Olmos, Dolores Velasco; Gastalum, Lourdes Cecilia Soto; Acosta, Magaly Aviles; Elizondo-Quiroga, Armando; Gonzalez-Rojas, Jose I.; Cordero, Juan F. Contreras; Guzman, Hilda; Travassos da Rosa, Amelia; Blitvich, Bradley J.; Barrett, Alan D.T.; Beaty, Barry J.

    2005-01-01

    West Nile virus has been isolated for the first time in Mexico, from a sick person and from mosquitoes (Culex quinquefasciatus). Partial sequencing and analysis of the 2 isolates indicate that they are genetically similar to other recent isolates from northern Mexico and the western United States. PMID:16229779

  16. Complete Genome Sequences of Five Zika Virus Isolates.

    PubMed

    Ladner, Jason T; Wiley, Michael R; Prieto, Karla; Yasuda, Chadwick Y; Nagle, Elyse; Kasper, Matthew R; Reyes, Daniel; Vasilakis, Nikolaos; Heang, Vireak; Weaver, Scott C; Haddow, Andrew; Tesh, Robert B; Sovann, Ly; Palacios, Gustavo

    2016-01-01

    Zika virus is an emerging human pathogen of great concern due to putative links to microcephaly and Guillain-Barre syndrome. Here, we report the complete genomes, including the 5' and 3' untranslated regions, of five Zika virus isolates, one from the Asian lineage and four from the African lineage. PMID:27174284

  17. Complete Genome Sequences of Five Zika Virus Isolates

    PubMed Central

    Ladner, Jason T.; Wiley, Michael R.; Prieto, Karla; Yasuda, Chadwick Y.; Nagle, Elyse; Kasper, Matthew R.; Reyes, Daniel; Vasilakis, Nikolaos; Heang, Vireak; Weaver, Scott C.; Haddow, Andrew; Tesh, Robert B.; Sovann, Ly

    2016-01-01

    Zika virus is an emerging human pathogen of great concern due to putative links to microcephaly and Guillain-Barre syndrome. Here, we report the complete genomes, including the 5′ and 3′ untranslated regions, of five Zika virus isolates, one from the Asian lineage and four from the African lineage. PMID:27174284

  18. Genome Sequences of Two Pseudorabies Virus Strains Isolated in Greece

    PubMed Central

    Papageorgiou, Konstantinos V.; Suárez, Nicolás M.; Wilkie, Gavin S.; Filioussis, George; Papaioannou, Nikolaos; Nauwynck, Hans J.; Kritas, Spyridon K.

    2016-01-01

    Pseudorabies virus (species Suid herpesvirus 1) belongs to the genus Varicellovirus, subfamily Alphaherpesvirinae, family Herpesviridae, and is the causative agent of an acute and frequently fatal disease that affects mainly pigs. Here, we report the genome sequences of two strains of this virus isolated in Greece in 2010. PMID:26798112

  19. Congenital rubella syndrome with positive serology and virus isolation.

    PubMed

    Ooi, H L; Cheong, S M; Yogeswery, S; Norizah, I; Zuridah, H; Kumarasamy, V; Chua, K B

    2006-06-01

    An effective live attenuated rubella vaccine was available since 1969 and congenital rubella syndrome can be prevented with appropriate vaccination. We report a baby with congenital rubella syndrome born in Klang valley to indicate that the Universal Rubella Vaccination Programme adopted by the Ministry of Health Malaysia since 2002 has yet to achieve its effect of eliminating transmission of rubella and preventing congenital rubella infection in the community. To our knowledge, the virus isolate represents the first successful isolation of rubella virus in this country and will serve as the reference strain for future comparison in molecular epidemiological tracking of rubella virus activity this country. PMID:16898324

  20. Detection and molecular characterization of Egyptian isolates of grapevine viruses.

    PubMed

    Fattouh, F; Ratti, C; El-Ahwany, A M D; Aleem, E Abdel; Babini, A R; Autonell, C Rubies

    2014-01-01

    Selected commercial and/or local vineyards and nurseries in three different governorates of Egypt (Alexandria, El-Beheira and El-Menofia) were surveyed for symptoms indicative of infection by grapevine viruses. Leaf samples from red-fruited and white-fruited Vitis vinefera were tested for grapevine leafroll associated viruses (GLRaV-1, GLRaV-2, and GLRaV-3), grapevine viruses A and B (GVA, GVB), grapevine rupestris stem pitting virus (GRSPaV), grapevine fanleaf virus (GFLV), and grapevine fleck virus (GFKV) from early April to late October 2010. Incidence of these viruses was assessed by RT-PCR in 60 different samples. Selected amplicons were sequenced. While GVA was the most wide spread (30%), GLRaV-1, GVB, GFLV, and GFKV were not detected during the survey. However, GVA, GLRaV-2, GLRaV-3, and GRSPaV were detected in the form of single infection or in mixed infections of 2 to 4 viruses. Phylogenetic analysis was performed on all Egyptian isolates of GLRaV-2 (4), GLRaV-3 (7), GVA (3), and GRSPaV (6). GRSPaV was detected for the first time in Egypt. Phylogenetic analysis provided insights into the evolutionary relationship between the reported Egyptian isolates and other previously reported isolates. PMID:24957718

  1. Updating strategies for isolating and discovering giant viruses.

    PubMed

    Khalil, Jacques Yaacoub Bou; Andreani, Julien; La Scola, Bernard

    2016-06-01

    Almost fifteen years ago, the discovery of Acanthamoeba polyphaga mimivirus, the first giant virus, changed how we define a virus. It was discovered incidentally in a process of isolating Legionella sp. from environmental samples in the context of pneumonia epidemics using a co-culture system with Acanthamoeba. Since then, much effort and improvement has been put into the original technique. In addition to the known families of Mimiviridae and Marseilleviridae, four new proposed families of giant viruses have been isolated: Pandoravirus, Pithovirus, Faustovirus and Mollivirus. Major improvements were based on enrichment systems, targeted use of antibiotics and high-throughput methods. The most recent development, using flow cytometry for isolation and presumptive identification systems, opens a path to large environmental surveys that may discover new giant virus families in new protozoa supports used for culture support. PMID:27039269

  2. Isolation of Genotype V St. Louis Encephalitis Virus in Florida

    PubMed Central

    Ambrose, Jason H.; White, Gregory S.; Unnasch, Thomas R.; Stark, Lillian M.

    2009-01-01

    We isolated and characterized St. Louis encephalitis virus (SLEV) from cloacal swabs of naturally exposed adult sentinel chickens in 2006. Phylogenetic analysis of SLEV strains isolated in Florida indicated that Brazilian SLEV circulated in 1972 and 2006; lineages were VA and VB. PMID:19331744

  3. West Nile Virus Isolation from Equines in Argentina, 2006

    PubMed Central

    Barrandeguy, María; Fabbri, Cintia; Garcia, Jorge B.; Vissani, Aldana; Trono, Karina; Gutierrez, Gerónimo; Pigretti, Santiago; Menchaca, Hernán; Garrido, Nelson; Taylor, Nora; Fernandez, Fernando; Levis, Silvana; Enría, Delia

    2006-01-01

    West Nile virus (WNV) was isolated from the brains of 3 horses that died from encephalitis in February 2006. The horses were from different farms in central Argentina and had not traveled outside the country. This is the first isolation of WNV in South America. PMID:17176571

  4. Variability in alternanthera mosaic virus isolates from different hosts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have determined the complete genome sequences of Alternanthera mosaic virus phlox isolate PA (AltMV-PA) and four infectious clone variants derived from AltMV-SP, as well as partial sequences of other isolates from various types of phlox, and from portulaca, nandina, and cineraria. Phylogenetic co...

  5. Isolation of influenza viruses in MDCK 33016PF cells and clearance of contaminating respiratory viruses.

    PubMed

    Roth, Bernhard; Mohr, Hannah; Enders, Martin; Garten, Wolfgang; Gregersen, Jens-Peter

    2012-01-11

    This paper summarizes results obtained by multiplex PCR screening of human clinical samples for respiratory viruses and corresponding data obtained after passaging of virus-positive samples in MDCK 33016PF cells. Using the ResPlexII v2.0 (Qiagen) multiplex PCR, 393 positive results were obtained in 468 clinical samples collected during an influenza season in Germany. The overall distribution of positive results was influenza A 42.0%, influenza B 38.7%, adenovirus 1.5%, bocavirus 0.5%, coronavirus 3.3%, enterovirus 5.6%, metapneumovirus 1.0%, parainfluenza virus 0.8%, rhinovirus 4.1%, and respiratory syncytial virus (RSV) 2.5%. Double infections of influenza virus together with another virus were found for adenovirus B and E, bocavirus, coronavirus, enterovirus and for rhinovirus. These other viruses were rapidly lost upon passages in MDCK 33016PF cells and under conditions as applied to influenza virus passaging. Clinical samples, in which no influenza virus but other viruses were found, were also subject to passages in MDCK 33016PF cells. Using lower inoculum dilutions than those normally applied for preparations containing influenza virus (total dilution of the original sample of ∼10(4)), the positive results for the different viruses turned negative already after 2 or 3 passages in MDCK 33016PF cells. These results demonstrate that, under practical conditions as applied to grow influenza viruses, contaminating viruses can be effectively removed by passages in MDCK cells. In combination with their superior isolation efficiency, MDCK cells appear highly suitable to be used as an alternative to embryonated eggs to isolate and propagate influenza vaccine candidate viruses. PMID:22119922

  6. Triticum Mosaic Virus: A New Virus Isolated From Wheat in Kansas

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In 2006 a mechanically-transmissible and previously uncharacterized virus was isolated in Kansas from wheat with mosaic symptoms. The physio-chemical properties of the virus were examined by purification on cesium chloride density gradients, electron microscopy, sodium dodecyl sulfate polyacrylalmid...

  7. Isolation of Naturally Occurring Viruses of the Murine Leukemia Virus Group in Tissue Culture

    PubMed Central

    Hartley, Janet W.; Rowe, Wallace P.; Capps, Worth I.; Huebner, Robert J.

    1969-01-01

    A tissue culture cell system for isolation and identification of members of the murine leukemia virus group (the complement fixation for murine leukemia test) was modified to permit the isolation of naturally occurring virus from leukemic and normal mice. The important factors for increasing the sensitivity of the test were the use of National Institutes of Health (NIH) strain Webster Swiss embryo cell cultures and the selection of rat-immune sera having complement-fixing antibodies to tissue culture antigens of both the Gross and FMR subgroups. In all, 163 strains of mouse leukemia virus, from 11 inbred mouse strains, have been isolated. Representative virus isolates were shown to possess the properties of the murine leukemia virus group; i.e., they were chloroform-sensitive, noncytopathic agents which replicated in mouse embryo tissue culture and produced group-reactive, complement-fixing antigen and budding C-type particles visible by electron microscopy. These viruses could serve as helpers in the rescue of Moloney sarcoma virus genome from non-producer hamster sarcoma cells, yielding pseudotypes. All of the 19 field isolates tested were neutralized by Gross passage A antiserum but not by potent antisera to the Moloney, Rauscher, and Friend strains. Virus was recovered regularly from embryos and from the plasma and spleen of adult mice of high leukemic strains. In low leukemic mouse strains, different patterns of virus detection were observed. In C3H/He mice, virus was occasionally present in embryos and was found in 40% of adult spleens. BALB/c mice were virus-negative as fetuses or weanlings, but spleens of more than half of the mice over 6 months of age yielded virus. NIH mice have never yielded virus. In reciprocal matings between AKR and BALB/c mice, virus recovery from embryos was maternally determined. The development of tissue culture isolation procedures made possible for the first time the application of classical infectious disease methods to the

  8. A Circo-Like Virus Isolated from Penaeus monodon Shrimps.

    PubMed

    Pham, Hanh T; Yu, Qian; Boisvert, Maude; Van, Hanh T; Bergoin, Max; Tijssen, Peter

    2014-01-01

    A virus with a circular Rep-encoding single-stranded DNA (ssDNA) (CRESS-DNA) genome (PmCV-1) was isolated from Penaeus monodon shrimps in Vietnam. The gene structure of the 1,777-nucleotide (nt) genome was similar to that of circoviruses and cycloviruses, but the nucleic acid and protein sequence identities to these viruses were very low. PMID:24435870

  9. Genetic typing of classical swine fever virus isolates from China.

    PubMed

    Sun, S-Q; Yin, S-H; Guo, H-C; Jin, Y; Shang, Y-J; Liu, X-T

    2013-08-01

    The E2 genes of 73 classical swine fever virus (CSFV) originated from CSF suspected cases in different regions of China were genetically characterized and compared with reference CSF viruses. All Chinese viruses that characterized were segregated into two major groups and subdivided into four subgroups. Most of isolates (61.6%) belonged to group 2 and were further divided into three subgroups: subgroup 2.1, 2.2 and 2.3. Subgroup 2.1 was the largest subgroup which contained 46.6% of isolates, while subgroup 2.3 was the smallest subgroup which contained only one isolate (1.4%). The remaining 38.4% of isolates were classified into subgroup 1.1 within group 1. However, no group 3 and subgroups 1.2 and 1.3 viruses were found in this study. This study has provided epidemiological information useful for assessing the virus origin and establishing a national prevention and control strategy against the disease. PMID:22672483

  10. Isolation of the envelope of vesicular stomatitis virus.

    PubMed Central

    Taube, S E; Rothfield, L I

    1978-01-01

    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

  11. Full Genomic Characterization of a Saffold Virus Isolated in Peru

    PubMed Central

    Leguia, Mariana; Loyola, Steev; Rios, Jane; Juarez, Diana; Guevara, Carolina; Silva, Maria; Prieto, Karla; Wiley, Michael; Kasper, Matthew R.; Palacios, Gustavo; Bausch, Daniel G.

    2015-01-01

    While studying respiratory infections of unknown etiology we detected Saffold virus in an oropharyngeal swab collected from a two-year-old female suffering from diarrhea and respiratory illness. The full viral genome recovered by deep sequencing showed 98% identity to a previously described Saffold strain isolated in Japan. Phylogenetic analysis confirmed the Peruvian Saffold strain belongs to genotype 3 and is most closely related to strains that have circulated in Asia. This is the first documented case report of Saffold virus in Peru and the only complete genomic characterization of a Saffold-3 isolate from the Americas. PMID:26610576

  12. Molecular and phenotypic characterization of infectious bursal disease virus isolates.

    PubMed

    Dormitorio, T V; Giambrone, J J; Guo, K; Jackwood, D J

    2007-06-01

    Two infectious bursal disease viruses (IBDVs 1174 and V1) were isolated from IBDV-vaccinated broiler flocks in California and Georgia. These flocks had a history of subclinical immunosuppression. These isolates are commonly used in IBDV progeny challenge studies at Auburn, AL, as well as vaccine manufacturer's vaccine efficacy studies, because they come from populated poultry-producing states, and are requested by poultry veterinarians from those states. Nested polymerase chain reaction (PCR) generated viral genome products for sequencing. A 491-bp segment from the VP2 gene, covering the hypervariable region, from each isolate was analyzed and compared with previously sequenced isolates. Sequence analysis showed that they were more closely related to the Delaware (Del) E antigenic variant than they are to the Animal Health Plant Inspection Service (APHIS) standard, both at the nucleotide level (96%, 97%) and at the amino acid level (94%, 97%). Both isolates had the glutamine to lysine shift in amino acid 249 which has been reported to be critical in binding the virus neutralizing Mab B69. Phenotypic studies showed that both isolates produced rapid atrophy of the bursae and weight loss, without the edematous bursal phase, in 2-wk-old commercial broilers having antibody against IBDV. A progeny challenge study showed both isolates produced more atrophy of the bursae (less percentage of protection) than the Del E isolate. Molecular and phenotypic data of these important IBDV isolates help in the improved detection and control of this continually changing and important viral pathogen of chickens. PMID:17626491

  13. Aphid Transmission of the Ontario Isolate of Plum Pox Virus.

    PubMed

    Lowery, D Thomas; Vickers, Patricia M; Bittner, Lori A; Stobbs, Lorne W; Foottit, Robert G

    2015-10-01

    Utilization of timed virus acquisition access probes in studies of plum pox virus (PPV) transmission by aphids demonstrated that endemic species transmitted the virus readily from plum, Prunus domestica (L.) Batsch; peach, P. persica (L.); or dwarf flowering almond, P. glandulosa Thunberg., to peach seedlings. The green peach aphid, Myzus persicae (Sulzer), was shown to be the most efficient vector. Acquisition of virus by green peach aphids from infected peach leaves resulted in 18-28% infected peach seedlings, while aphids previously fed on infected leaves of plum transferred virus to 36% of peach seedlings. Although the spirea aphid, Aphis spiraecola (Patch), was a less efficient vector than M. persicae it is perhaps more important for the spread of PPV due to its greater abundance and occurrence earlier in the season when peach trees are thought to be more susceptible to infection. Virus transmission rates varied depending on the virus source and healthy test plant species. In contrast to many previous studies, aphid inoculation of the experimental host Nicotiana benthamiana Domin occurred at a low rate, never exceeding 4%. Acquisition of PPV by M. persicae from infected peach fruit was greatly reduced compared with acquisition from leaves. The results of this research indicate that the Ontario isolate of PPV-D is readily transmissible by aphids to peach and natural spread of the virus needs to be considered in future management or eradication programs. PMID:26453705

  14. Characterization and phylogenic analysis of Mexican Newcastle disease virus isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Newcastle disease virus (NDV) was isolated in Mexico for the first time in 1946 and the last report of a field outbreak caused by a highly virulent strain dates from year 2000, when 13.6 million birds were slaughtered and 93 farms quarantined. Mean Death Time test resulted in velogenic classificati...

  15. Concurrent Isolation of Chikungunya Virus and Dengue Virus from a Patient with Coinfection Resulting from a Trip to Singapore ▿

    PubMed Central

    Chang, Shu-Fen; Su, Chien-Ling; Shu, Pei-Yun; Yang, Cheng-Fen; Liao, Tsai-Ling; Cheng, Chia-Hsin; Hu, Huai-Chin; Huang, Jyh-Hsiung

    2010-01-01

    We report two cases of imported infection in patients who had returned to Taiwan from Singapore: one was coinfected with chikungunya virus and dengue virus type 2, and the other was infected with the same dengue virus. Both viruses were successfully isolated from the coinfected case by using antibody neutralization and a plaque purification technique. PMID:20881182

  16. Isolation and characterization of an H9N2 influenza virus isolated in Argentina

    PubMed Central

    Xu, Kemin; Ferreri, Lucas; Rimondi, Agustina; Olivera, Valeria; Romano, Marcelo; Ferreyra, Hebe; Rago, Virgina; Uhart, Marcela; Chen, Hongjun; Sutton, Troy; Pereda, Ariel; Perez, Daniel R.

    2016-01-01

    As part of our ongoing efforts on animal influenza surveillance in Argentina, an H9N2 virus was isolated from a wild aquatic bird (Netta peposaca), A/rosy-billed pochard/Argentina/CIP051-559/2007 (H9N2) – herein referred to as 559/H9N2. Due to the important role that H9N2 viruses play in the ecology of influenza in nature, the 559/H9N2 isolate was characterized molecularly and biologically. Phylogenetic analysis of the HA gene revealed that the 559/H9N2 virus maintained an independent evolutionary pathway and shared a sister-group relationship with North American viruses, suggesting a common ancestor. The rest of the genome segments clustered with viruses from South America. Experimental inoculation of the 559/H9N2 in chickens and quail revealed efficient replication and transmission only in quail. Our results add to the notion of the unique evolutionary trend of avian influenza viruses in South America. Our study increases our understanding of H9N2 viruses in nature and emphasizes the importance of expanding animal influenza surveillance efforts to better define the ecology of influenza viruses at a global scale. PMID:22709552

  17. Molecular characterization of rabies virus isolates from Trinidad.

    PubMed

    Wright, Arlette; Rampersad, Joanne; Ryan, Joseph; Ammons, David

    2002-06-20

    Bovine rabies continues to be a serious problem facing the cattle industry in South and Central America. Although Trinidad played an important role in originally demonstrating the link between bats and bovine rabies, relatively little is known about rabies in Trinidad, an island 7miles off the coast of Venezuela. In order to obtain a more complete understanding of bovine rabies in the region, we report herein on a study undertaken in Trinidad to characterize isolates of rabies virus obtained from infected cattle. A portion of the nucleotide sequence of the nucleoprotein gene from six rabies virus isolates collected from bovine rabies from the years 1997, 1998 and 2000 was determined and compared both to themselves and the nucleotide sequence of other South American isolates. Results indicate that there are at least two independently evolving variants of rabies virus in Trinidad. The nucleotide sequence of either variant failed to match completely the sequence of South American isolates. However, the lack of South American isolates from coastal regions facing Trinidad leaves undetermined the question of South American influence on rabies in Trinidad. The results of this study helps complete the picture of bovine rabies in the South American region and provide basic information required locally for the creation of an effective rabies control and eradication strategy. PMID:12034537

  18. An evolutionary insight into Newcastle disease viruses isolated in Antarctica.

    PubMed

    Soñora, Martin; Moreno, Pilar; Echeverría, Natalia; Fischer, Sabrina; Comas, Victoria; Fajardo, Alvaro; Cristina, Juan

    2015-08-01

    The disease caused by Newcastle disease virus (NDV) is a severe threat to the poultry industry worldwide. Recently, NDV has been isolated in the Antarctic region. Detailed studies on the mode of evolution of NDV strains isolated worldwide are relevant for our understanding of the evolutionary history of NDV. For this reason, we have performed Bayesian coalescent analysis of NDV strains isolated in Antarctica to study evolutionary rates, population dynamics, and patterns of evolution. Analysis of F protein cleavage-site sequences of NDV isolates from Antarctica suggested that these strains are lentogenic. Strains isolated in Antarctica and genotype I reference strain Ulster/67 diverged from ancestors that existed around 1958. The time of the most recent common ancestor (MRCA) was established to be around 1883 for all class II viruses. A mean rate of evolution of 1.78 × 10(-3) substitutions per site per year (s/s/y) was obtained for the F gene sequences of NDV strains examined in this study. A Bayesian skyline plot indicated a decline in NDV population size in the last 25 years. The results are discussed in terms of the possible role of Antarctica in emerging or re-emerging viruses and the evolution of NDV populations worldwide. PMID:26014920

  19. Isolation and Characterization of an Equine Foamy Virus

    PubMed Central

    Tobaly-Tapiero, Joelle; Bittoun, Patricia; Neves, Manuel; Guillemin, Marie-Claude; Lecellier, Charles-Henri; Puvion-Dutilleul, Francine; Gicquel, Bernard; Zientara, Stephan; Giron, Marie-Louise; de Thé, Hugues; Saïb, Ali

    2000-01-01

    Foamy viruses (FVs) are complex retroviruses which have been isolated from different animal species including nonhuman primates, cattle, and cats. Here, we report the isolation and characterization of a new FV isolated from blood samples of horses. Similar to other FVs, the equine foamy virus (EFV) exhibits a highly characteristic ultrastructure and induces syncytium formation and subsequent cell lysis on a large number of cell lines. Molecular cloning of EFV reveals that the general organization is that of other known FVs, whereas sequence similarity with its bovine FV counterpart is only 40%. Interestingly, EFV buds exclusively from the plasma membrane and not from the endoplasmic reticulum (ER), as previously shown for other FVs. The absence of the ER retrieval dilysine motif in EFV Env is likely responsible for this unexpected sorting pathway. PMID:10756018

  20. Mayaro virus fever in French Guiana: isolation, identification, and seroprevalence.

    PubMed

    Talarmin, A; Chandler, L J; Kazanji, M; de Thoisy, B; Debon, P; Lelarge, J; Labeau, B; Bourreau, E; Vié, J C; Shope, R E; Sarthou, J L

    1998-09-01

    This paper reports the first isolation of Mayaro (MAY) virus from a patient infected in French Guiana. The identification was initially performed using immunofluorescent antibody testing with specific mouse antibody, and confirmed by plaque-reduction neutralization testing and reverse transcription-polymerase chain reaction. To determine if MAY virus infection is widespread in French Guiana, a serosurvey was performed to determine the prevalence of antibody to this virus in various ethnic groups and areas of French Guiana. Human sera (n = 1,962) were screened using the hemagglutination inhibition (HI) test. To determine whether MAY virus circulates in the rain forest, a serosurvey in monkey populations was performed. Monkey sera (n = 150) were also screened for antibody to MAY virus using HI testing. Of the human sera tested, 6.3% were positive for anti-MAY virus antibodies. Significant differences in MAY virus seroprevalence between different age groups were observed. Seroprevalence rates increased with age, with a large increase in people 10-19 years of age in comparison with those less than 10 years of age. After adjustment for age, significant differences were also found between places of residence. The prevalence of anti-MAY virus antibody was higher in people living in contact with the forest, especially in the Haut Oyapock area (odds ratio [OR] = 97.7, 95% confidence interval [CI] = 48.2-197.9) and along the Maroni River (OR = 39.7, 95% CI = 20.6-76.6). The ethnic differences observed in this study were probably due to differences in residence. Among monkeys, higher seroprevalence rates were found in Alouatta seniculus (66.0%) than in Saguinus midas (18.2%). Among Alouatta, the seroprevalence increased significantly with weight (and therefore with age). This study indicates that MAY virus is present in French Guiana, and human infections occur in areas where people live near the tropical rain forest. PMID:9749643

  1. A novel totivirus-like virus isolated from bat guano.

    PubMed

    Yang, Xinglou; Zhang, Yunzhi; Ge, Xingyi; Yuan, Junfa; Shi, Zhengli

    2012-06-01

    Previous metagenomic analysis indicated that numerous insect viruses exist in bat guano. In this study, we isolated a novel double-stranded RNA virus, a tentative member of the family Totiviridae, designated Tianjin totivirus (ToV-TJ), from bat feces. The virus is an icosahedral particle with a diameter of 40-43 nm, and it causes cytopathic effect in Sf9, Hz, and C6/36 cell lines. Full-length genomic sequence analysis showed that ToV-TJ shares high similarity with the totivirus OMRV-AK4, which was recently isolated from mosquitoes in Japan. The full-length genome of the ToV-TJ was 7611 bp and contained two predicted non-overlapping open reading frames (ORFs): ORF1, encoding the capsid protein (CP), and ORF2, encoding an RNA-dependent RNA polymerase. Bioassay of ToV-TJ by feeding on the larvae of Spodoptera exigua and Helicoverpa armigera (Hubner) suggests that this virus is not infectious for these two larvae in vivo. Sequences similar to that of ToV-TJ have been detected in bat feces sampled in Yunnan and Hainan Provinces, suggesting that this virus is widely distributed. PMID:22421961

  2. Recent isolations of Lassa virus from Nigerian rodents

    PubMed Central

    Wulff, Herta; Fabiyi, A.; Monath, T. P.

    1975-01-01

    Rodents were trapped in the Benue-Plateau and North-Eastern States of Nigeria where Lassa fever had been reported in previous years. Eight Lassa virus strains were isolated from tissues and blood of rodents identified in the field as being of 3 different species: Mastomys natalensis, Rattus rattus, and Mus minutoides. All the infected rodents were collected in village habitats. These isolations indicate the presence of Lassa virus in wild rodents in Nigeria during periods when no human infections were evident. Prior studies in Sierra Leone have indicated that a single rodent species, M. natalensis, may be the important reservoir host of Lassa virus. Since the present study indicates that other rodent species may be involved as well, the ecology of Lassa virus may be more complicated than was heretofore supposed. In view of the importance of determining the geographic and species range of rodent hosts of Lassa virus, and because of the problems inherent in rodent identification under austere field conditions, it is urgent that further studies be conducted in the same areas of Nigeria to confirm these findings. PMID:1085216

  3. Amplification of 'variola virus-specific' sequences in German cowpox virus isolates.

    PubMed

    Meyer, H; Neubauer, H; Pfeffer, M

    2002-02-01

    In 1995 a polymerase chain reaction (PCR) protocol describing the specific detection of variola virus, the causative agent of smallpox, was published by Knight and others. Virulent variola major strains could be differentiated from less virulent variola minor strains because of the distinct amplicon sizes. Here, we applied this PCR protocol to DNA from various orthopoxvirus isolates. There was no amplification with the orthopoxvirus species vaccinia, monkeypox, mousepox, or camelpox viruses. However, amplification was observed in six out of 15 cowpox virus strains investigated. The size of the amplicons corresponded exactly with the size described for variola minor strains and the nucleotide sequence identity accounted for 97%. Findings are discussed with respect to the evolution of orthopoxvirus species assuming that variola virus most probably stems from a rodent-transmitted cowpox virus-like progenitor. PMID:11911586

  4. Isolation and molecular characterization of Newcastle disease viruses from raptors.

    PubMed

    Jindal, Naresh; Chander, Yogesh; Primus, Alexander; Redig, Patrick T; Goyal, Sagar M

    2010-12-01

    The present study was undertaken to detect and characterize Newcastle disease virus (NDV) in raptors. Cloacal and oropharyngeal swab samples were collected from 60 casualty raptors during January to March 2009 in Minnesota. Inoculation of all these samples (n=120) in 9-day-old embryonated hens' eggs resulted in isolation of haemagglutinating viruses in three samples from two bald eagles and one great horned owl. These three haemagglutinating viruses were confirmed as NDV by reverse transcription-polymerase chain reaction (RT-PCR) using fusion gene-specific primers, and were negative for avian influenza virus by RT-PCR. Further characterization revealed that all three possessed (112)GKQGRL(117) at the fusion gene cleavage site, indicating that they were lentogenic strains. Phylogenetic analysis revealed that all three isolates clustered with published class II genotype II NDVs. The nucleotide sequence homology of the three NDV isolates among themselves was 98.4 to 99.6% and the sequence homology with lentogenic strains from wild birds used for comparison varied between 94.5 and 100%. Detection of NDV strains from raptors merits further epidemiological studies to determine the prevalence of different NDV strains in raptors and their impact in relation to transmission to domestic poultry. PMID:21154052

  5. Isolation, pathogenicity, and H120 protection efficacy of infectious bronchitis viruses isolated in Taiwan.

    PubMed

    Wang, C H; Hsieh, M C; Chang, P C

    1996-01-01

    Seven isolates of infectious bronchitis (IB) virus (IBV) were isolated from two breeder farms and five broiler farms in Taiwan in 1992. The cardinal signs of disease in breeders were egg production drops and watery albumen, and those in broilers were respiratory distress and renal urate deposition or death. All diseased chickens had been vaccinated with IB vaccines (mostly H120). The viruses were isolated and identified by chicken embryo inoculation and electron microscopy. The genomes of the isolates were extracted and amplified by polymerase chain reaction; the restriction fragment length polymorphism analysis suggested that the genotypes of the present IBV isolates were different from the eight reference strains. One-day-old specific-pathogen-free chicks were inoculated with the field isolates in order to test the virulence of those isolates. Respiratory distress and depression commenced at 24 hours after inoculation. Two of the seven isolates were found to be highly virulent, causing 50% or more mortality in inoculated chicks. Vaccine protection tests showed that H120 could protect chickens against challenges with four of six field isolates. PMID:8883793

  6. Suppression of influenza virus infection by the orf virus isolated in Taiwan

    PubMed Central

    LIN, Fong-Yuan; TSENG, Yeu-Yang; CHAN, Kun-Wei; KUO, Shu-Ting; YANG, Cheng-Hsiung; WANG, Chi-Young; TAKASU, Masaki; HSU, Wei-Li; WONG, Min-Liang

    2015-01-01

    Orf virus (ORFV), a member of parapoxvirus, is an enveloped virus with genome of double-stranded DNA. ORFV causes contagious pustular dermatitis or contagious ecthyma in sheep and goats worldwide. In general, detection of viral DNA and observing ORFV virion in tissues of afflicted animals are two methods commonly used for diagnosis of orf infection; however, isolation of the ORFV in cell culture using virus-containing tissue as inoculum is known to be difficult. In this work, the ORFV (Hoping strain) isolated in central Taiwan was successfully grown in cell culture. We further examined the biochemical characteristic of our isolate, including viral genotyping, viral mRNA and protein expression. By electron microscopy, one unique form of viral particle from ORFV infected cellular lysate was demonstrated in the negative-stained field. Moreover, immunomodulating and anti-influenza virus properties of this ORFV were investigated. ORFV stimulated human monocytes (THP-1) secreting proinflammatory cytokines IL-8 and TNF-α. And, pre-treatment of ORFV-infected cell medium prevents A549 cells from subsequent type A influenza virus (IAV) infection. Similarly, mice infected with ORFV via both intramuscular and subcutaneous routes at two days prior to IAV infection significantly decreased the replication of IAV. In summary, the results of a current study indicated our Hoping strain harbors the immune modulator property; with such a bio-adjuvanticity, we further proved that pre-exposure of ORFV protects animals from subsequent IAV infection. PMID:25855509

  7. Comparison of Akabane virus isolated from sentinel cattle in Japan.

    PubMed

    Ogawa, Yohsuke; Fukutomi, Toyoko; Sugiura, Keita; Sugiura, Katsuaki; Kato, Kentaro; Tohya, Yukinobu; Akashi, Hiroomi

    2007-09-20

    Adult cows, ewes, and goats infected with Akabane virus (AKAV) of the genus Orthobunyavirus of the family Bunyaviridae do not present any clinical signs; however, in utero infections may result in abortion, premature birth, stillbirth, and congenital deformities such as arthrogryposis-hydranencephaly syndrome in cattle, sheep, and goats. In contrast, the Iriki strain, a variant of AKAV isolated from a calf with nervous signs and encephalitis, causes encephalitis in experimentally inoculated calves. Two AKAV field isolates, named Okayama2001 and Okayama2004, were isolated from blood specimens of sentinel calves and characterized by cross-neutralization testing, genetic analyses of the S and M RNA segments, and experimental intraperitoneal infection in mice. Although a genetic relationship was established between Okayama2001 and the Iriki strain, their antigenic characteristics differ. Okayama2001 was avirulent in mice, as was the OBE-1 strain, which was isolated from an aborted bovine fetus. In contrast, Okayama2004 was antigenically and genetically related to the OBE-1 strain, but was virulent in mice, similar to the Iriki strain. These results indicate that the isolates mutated antigenically or pathogenically and suggest that AKAV mutates frequently in the field. Although attenuated and inactivated vaccines have been developed for disease prevention, an outbreak may occur due to variant viruses arising from mutation. PMID:17467929

  8. Isolation and phylogenetic analysis of caprine Orf virus in Malaysia.

    PubMed

    Abdullah, Ashwaq Ahmed; Ismail, Muhammad Farid Bin; Balakrishnan, Krishnan Nair; Bala, Jamilu Abubakar; Hani, Homayoun; Abba, Yusuf; Awang Isa, Mohd Kamaruddin; Abdullah, Faez Firdaus Jesse; Arshad, Siti Suri; Nazariah, Zeenatul Allaudin; Abdullah, Rasedee; Mustapha, Noordin Mohamed; Mohd-Lila, Mohd-Azmi

    2015-12-01

    Orf virus is a DNA virus that causes contiguous ecthyma in goat and sheep. Infection of animals with this virus cause high mortality in young animals resulting in huge economic losses. In this study, we investigated an outbreak of Orf in a goat farm in Malaysia. Samples were collected from infected animals and viral isolation was done using both LT and MDCK cell lines. Molecular detection was done by conventional PCR for specific primers; B2L and F1L genes and phylogenetic analysis was done on the sequence data obtained. Cytopathic effects (CPE) were observed in both cell lines after 3 days of inoculation and were 50 % by the sixth day. PCR showed positive bands for both B2L and F1L genes and phylogenetic analysis showed that the Malaysian strain had close homology to the Chinese and Indian Orf virus isolates. This study gives more insight into the existing Orf viral strains in Malaysia and their relationship with other strains globally. PMID:26645035

  9. Molecular characterisation of virulence graded field isolates of myxoma virus

    PubMed Central

    2010-01-01

    Background Myxoma virus (MV) has been endemic in Europe since shortly after its deliberate release in France in 1952. While the emergence of more resistant hosts and more transmissible and attenuated virus is well documented, there have been relatively few studies focused on the sequence changes incurred by the virus as it has adapted to its new host. In order to identify regions of variability within the MV genome to be used for phylogenetic studies and to try to investigate causes of MV strain attenuation we have molecularly characterised nine strains of MV isolated in Spain between the years 1992 and 1995 from wide ranging geographic locations and which had been previously graded for virulence by experimental infection of rabbits. Results The findings reported here show the analysis of 16 genomic regions accounting for approximately 10% of the viral genomes. Of the 20 genes analysed 5 (M034L, M069L, M071L, M130R and M135R) were identical in all strains and 1 (M122R) contained only a single point mutation in an individual strain. Four genes (M002L/R, M009L, M036L and M017L) showed insertions or deletions that led to disruption of the ORFs. Conclusions The findings presented here provide valuable tools for strain differentiation and phylogenetic studies of MV isolates and some clues as to the reasons for virus attenuation in the field. PMID:20187925

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

    PubMed Central

    Martins, Lívia C.; Diniz Júnior, José Antonio P.; de Almeida Medeiros, Daniele Barbosa; Cardoso, Jedson F.; da Silva, Daisy E. A.; de Oliveira, Layanna F.; de Vasconcelos, Janaina M.; Vianez Júnior, João Lídio da S. G.; Vasconcelos, Pedro F. C.

    2014-01-01

    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

  11. The E116 isolate of Dutch pea early-browning virus is a recombinant virus.

    PubMed

    Swanson, M M; MacFarlane, S A

    1999-03-01

    The complete nucleotide sequence of RNA2 of the E116 isolate of Dutch pea early-browning virus (PEBV-D) was obtained from overlapping cDNA clones. The RNA was found to encode three open reading frames corresponding to, in 5' to 3' order, the coat protein, the 2b nematode transmission protein and the C-terminal part of the cysteine-rich 1b protein derived from RNA1. The 3' non-coding region of PEBV-D RNA2 was also shown to be derived from RNA1. This is the first demonstration that recombination of PEBV occurs in nature. Comparison of the amino acid sequences of the PEBV-D RNA2 proteins with those of British PEBV and several isolates of tobacco rattle virus reveals complex patterns of mixing of the genomes of these two viruses. PMID:10225277

  12. Comparative study of different isolates of murine sarcoma virus.

    PubMed Central

    Donoghue, D J; Sharp, P A; Weinberg, R A

    1979-01-01

    The RNA genomes of a variety of murine sarcoma viruses (MSV) were compared by heteroduplex analysis. These viruses included the Moloney-derived isolates 124-MSV, m1-MSV, m3-MSV, HT1-MSV, and NP-MSV and also two independent isolates, Gazdar MSV and 1712-MSV. All of these viral genomes exhibited the acquired cellular sequences previously identified in 3124-MSV and thought to be responsible for transformation and sarcomagenesis. The location of the acquired cellular sequences within the envelope gene was variable in different MSV isolates, suggesting that the cellular sequences can be expressed in different positions relative to murine leukemia virus-derived information present in MSV. Deletions in the gag coding region of the different MSVs were consistent with their known gag-related gene products. Based on several features of the hetero-duplex analysis and the known genealogical relationships of the different MSVs, various possible mechanisms for the formation of MSV are considered. Images PMID:229256

  13. Antigenic and genetic characterization of rabies virus isolates from Uruguay.

    PubMed

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

    2013-05-01

    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

  14. Isolation and genetic characterization of a tembusu virus strain isolated from mosquitoes in Shandong, China.

    PubMed

    Tang, Y; Diao, Y; Chen, H; Ou, Q; Liu, X; Gao, X; Yu, C; Wang, L

    2015-04-01

    Tembusu virus (TMUV) is a flavivirus, presumed to be a mosquito-borne flavivirus of the Ntaya virus subgroup. To date, however, there have been no reports indicating that mosquitoes are involved in the spread of TMUV. In this study, we report the first isolation of TMUV from Culex mosquitoes. We describe the isolation and characterization of a field strain of TMUV from mosquitoes collected in Shandong Province, China. The virus isolate, named TMUV-SDMS, grows well in mosquito cell line C6/36, in Vero and duck embryo fibroblast (DEF) cell lines, and causes significant cytopathic effects in these cell cultures. The TMUV-SDMS genome is a single-stranded RNA, 10 989 nt in length, consisting of a single open reading frame encoding a polyprotein of 3410 amino acids, with 5' and 3' untranslated regions of 142 and 617 nt, respectively. Phylogenetic analysis of the E and NS5 genes revealed that the TMUV-SDMS is closely related to the TMUV YY5 and BYD strains which cause severe egg-drop in ducks. The 3'NTR of TMUV-SDMS contains two pairs of tandem repeat CS and one non-duplicate CS, which have sequence similarities to the same repeats in the YY5 and BYD strains. Our findings indicate that mosquitoes carrying the TMUV may play an important role in the spread of this virus and in disease outbreak. PMID:23711093

  15. Cedar Virus: A Novel Henipavirus Isolated from Australian Bats

    PubMed Central

    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

    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-β response than HeV. PMID:22879820

  16. Genomic characterization of pseudorabies virus strains isolated in Italy.

    PubMed

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

    2014-08-01

    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

  17. Phylogenetic and serological analysis of turnip ringspot virus and radish mosaic virus isolates.

    PubMed

    Koloniuk, Igor; Petrzik, Karel

    2012-03-01

    Turnip ringspot virus (TuRSV) has been proposed to be a member of a new species in the genus Comovirus. Its remarkable host-range similarity to radish mosaic virus (RaMV) may have led to its misrecognition in the past. Findings from both sequence analysis and serological tests support the assignment of TuRSV to a new comovirus species. In addition, phylogenetic analysis suggests that the two genome segments of some TuRSV isolates have a heterogeneous origin. PMID:22160585

  18. Evaluation of rabies biologics against Irkut virus isolated in China.

    PubMed

    Liu, Ye; Chen, Qi; Zhang, Fei; Zhang, Shoufeng; Li, Nan; Lian, Hai; Wang, Ying; Zhang, Jinxia; Hu, Rongliang

    2013-11-01

    An Irkut virus (IRKV) was recently isolated from a bat in China. The protective ability of rabies biologics available in the Chinese market and experimental biologics against the rabies virus (RABV) and IRKV were assessed in a hamster model via preexposure prophylaxis (PrEP) and postexposure prophylaxis (PEP) experiments. The results demonstrated that a single dose of rabies vaccine did not induce adequate protection against IRKV infection. However, routine PrEP with three doses of vaccine induced complete protection against IRKV infection. Higher doses of RABV immunoglobulins and alpha interferon were required during PEP to protect hamsters against IRKV versus RABV infection. Experimental recombinant vaccines containing IRKV glycoproteins induced more-reliable protection against IRKV than against RABV infection. Those findings may be explained by limited cross-neutralization of these viruses (confirmed via in vitro tests) in conjunction with antigenic distances between RABV and IRKV. These results indicate that the development and evaluation of new biologics for PrEP and PEP are required to ensure sufficient protection against IRKV infection in China and other territories where this virus is present. PMID:23946522

  19. Complete Genome Sequence of Tomato Mosaic Virus Isolated from Jasmine in the United States.

    PubMed

    Fillmer, Kornelia; Adkins, Scott; Pongam, Patchara; D'Elia, Tom

    2015-01-01

    Tomato mosaic virus was reported from jasmine in Florida. We present the first complete genome sequence of a tomato mosaic virus isolate from this woody perennial plant in the United States. PMID:26159525

  20. First Complete Genome Sequence of a Watermelon Mosaic Virus Isolated from Watermelon in the United States

    PubMed Central

    Rajbanshi, Naveen

    2016-01-01

    Watermelon mosaic virus was first reported in 1965 from the Rio Grande Valley, TX. We report here the first complete genome sequence of a watermelon mosaic virus isolate from watermelon collected from the Rio Grande Valley of Texas. PMID:27103724

  1. First Complete Genome Sequence of a Watermelon Mosaic Virus Isolated from Watermelon in the United States.

    PubMed

    Rajbanshi, Naveen; Ali, Akhtar

    2016-01-01

    Watermelon mosaic virus was first reported in 1965 from the Rio Grande Valley, TX. We report here the first complete genome sequence of a watermelon mosaic virus isolate from watermelon collected from the Rio Grande Valley of Texas. PMID:27103724

  2. Isolation, Genetic Characterization, and Seroprevalence of Adana Virus, a Novel Phlebovirus Belonging to the Salehabad Virus Complex, in Turkey

    PubMed Central

    Alkan, Cigdem; Alwassouf, Sulaf; Piorkowski, Géraldine; Bichaud, Laurence; Tezcan, Seda; Dincer, Ender; Ergunay, Koray; Ozbel, Yusuf; Alten, Bulent; de Lamballerie, Xavier

    2015-01-01

    ABSTRACT A new phlebovirus, Adana virus, was isolated from a pool of Phlebotomus spp. (Diptera; Psychodidae) in the province of Adana, in the Mediterranean region of Turkey. Genetic analysis based on complete coding of genomic sequences indicated that Adana virus belongs to the Salehabad virus species of the genus Phlebovirus in the family Bunyaviridae. Adana virus is the third virus of the Salehabad virus species for which the complete sequence has been determined. To understand the epidemiology of Adana virus, a seroprevalence study using microneutralization assay was performed to detect the presence of specific antibodies in human and domestic animal sera collected in Adana as well as Mersin province, located 147 km west of Adana. The results demonstrate that the virus is present in both provinces. High seroprevalence rates in goats, sheep, and dogs support intensive exposure to Adana virus in the region, which has not been previously reported for any virus included in the Salehabad serocomplex; however, low seroprevalence rates in humans suggest that Adana virus is not likely to constitute an important public health problem in exposed human populations, but this deserves further studies. IMPORTANCE Until recently, in the genus Phlebovirus, the Salehabad virus species consisted of two viruses: Salehabad virus, isolated from sand flies in Iran, and Arbia virus, isolated from sand flies in Italy. Here we present the isolation and complete genome characterization of the Adana virus, which we propose to be included in the Salehabad virus species. To our knowledge, this is the first report of the isolation and complete genome characterization, from sand flies in Turkey, of a Salehabad virus-related phlebovirus with supporting seropositivity in the Mediterranean, Aegean, and Central Anatolia regions, where phleboviruses have been circulating and causing outbreaks. Salehabad species viruses have generally been considered to be a group of viruses with little medical or

  3. Complete Genome Sequences of Two Hydrangea Ringspot Virus Isolates from Japan

    PubMed Central

    Yusa, Akira; Iwabuchi, Nozomu; Koinuma, Hiroaki; Keima, Takuya; Neriya, Yutaro; Hashimoto, Masayoshi; Maejima, Kensaku; Yamaji, Yasuyuki

    2016-01-01

    Hydrangea ringspot virus (HdRSV) is a plant RNA virus, naturally infecting Hydrangea macrophylla. Here, we report the first genomic sequences of two HdRSV isolates from hydrangea plants in Japan. The overall nucleotide sequences of these Japanese isolates were 96.0 to 96.3% identical to those of known European isolates. PMID:27034476

  4. Application of the "best fit" pathotyping assay for evaluation of Russian isolates of Marek's disease virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to investigate the validity of the "best fit" pathotyping assay for testing of field isolates of Marek's disease (MD) virus. Twenty serotype 1 MD viruses were isolated from breeder and commercial flocks in 8 regions of the Russian Federation. These isolates were pat...

  5. Characterization of a Siberian Virus Isolated from a Patient with Progressive Chronic Tick-Borne Encephalitis

    PubMed Central

    Gritsun, T. S.; Frolova, T. V.; Zhankov, A. I.; Armesto, M.; Turner, S. L.; Frolova, M. P.; Pogodina, V. V.; Lashkevich, V. A.; Gould, E. A.

    2003-01-01

    A strain of Tick-borne encephalitis virus designated Zausaev (Za) was isolated in Siberia from a patient who died of a progressive (2-year) form of tick-borne encephalitis 10 years after being bitten by a tick. The complete genomic sequence of this virus was determined, and an attempt was made to correlate the sequence with the biological characteristics of the virus. Phylogenetic analysis demonstrated that this virus belongs to the Siberian subtype of Tick-borne encephalitis virus. Comparison of Za virus with two related viruses, a Far Eastern isolate, Sofjin, and a Siberian isolate, Vasilchenko, revealed differences among the three viruses in pathogenicity for Syrian hamsters, cytopathogenicity for PS cells, plaque morphology, and the electrophoretic profiles of virus-specific nonstructural proteins. Comparative amino acid alignments revealed 10 individual amino acid substitutions in the Za virus polyprotein sequence that were different from those of other tick-borne flaviviruses. Notably, the dimeric form of the Za virus NS1 protein migrated in polyacrylamide gels as a heterogeneous group of molecules with a significantly higher electrophoretic mobility than those of the Sofjin and Vasilchenko viruses. Two amino acid substitutions, T277→V and E279→G, within the NS1 dimerization domain are probably responsible for the altered oligomerization of Za virus NS1. These studies suggest that the patient from whom Za virus was isolated died due to increased pathogenicity of the latent virus following spontaneous mutagenesis. PMID:12477807

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

    PubMed Central

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

    2013-01-01

    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

  7. Virus fitness differences observed between two naturally occurring isolates of Ebola virus Makona variant using a reverse genetics approach.

    PubMed

    Albariño, César G; Guerrero, Lisa Wiggleton; Chakrabarti, Ayan K; Kainulainen, Markus H; Whitmer, Shannon L M; Welch, Stephen R; Nichol, Stuart T

    2016-09-01

    During the large outbreak of Ebola virus disease that occurred in Western Africa from late 2013 to early 2016, several hundred Ebola virus (EBOV) genomes have been sequenced and the virus genetic drift analyzed. In a previous report, we described an efficient reverse genetics system designed to generate recombinant EBOV based on a Makona variant isolate obtained in 2014. Using this system, we characterized the replication and fitness of 2 isolates of the Makona variant. These virus isolates are nearly identical at the genetic level, but have single amino acid differences in the VP30 and L proteins. The potential effects of these differences were tested using minigenomes and recombinant viruses. The results obtained with this approach are consistent with the role of VP30 and L as components of the EBOV RNA replication machinery. Moreover, the 2 isolates exhibited clear fitness differences in competitive growth assays. PMID:27366976

  8. First isolation of West Nile virus from a dromedary camel

    PubMed Central

    Joseph, Sunitha; Wernery, Ulrich; Teng, Jade LL; Wernery, Renate; Huang, Yi; Patteril, Nissy AG; Chan, Kwok-Hung; Elizabeth, Shyna K; Fan, Rachel YY; Lau, Susanna KP; Kinne, Jörg; Woo, Patrick CY

    2016-01-01

    Although antibodies against West Nile virus (WNV) have been detected in the sera of dromedaries in the Middle East, North Africa and Spain, no WNV has been isolated or amplified from dromedary or Bactrian camels. In this study, WNV was isolated from Vero cells inoculated with both nasal swab and pooled trachea/lung samples from a dromedary calf in Dubai. Complete-genome sequencing and phylogenetic analysis using the near-whole-genome polyprotein revealed that the virus belonged to lineage 1a. There was no clustering of the present WNV with other WNVs isolated in other parts of the Middle East. Within lineage 1a, the dromedary WNV occupied a unique position, although it was most closely related to other WNVs of cluster 2. Comparative analysis revealed that the putative E protein encoded by the genome possessed the original WNV E protein glycosylation motif NYS at E154–156, which contained the N-linked glycosylation site at N-154 associated with increased WNV pathogenicity and neuroinvasiveness. In the putative NS1 protein, the A70S substitution observed in other cluster 2 WNVs and P250, which has been implicated in neuroinvasiveness, were present. In addition, the foo motif in the putative NS2A protein, which has been implicated in neuroinvasiveness, was detected. Notably, the amino-acid residues at 14 positions in the present dromedary WNV genome differed from those in most of the closely related WNV strains in cluster 2 of lineage 1a, with the majority of these differences observed in the putative E and NS5 proteins. The present study is the first to demonstrate the isolation of WNV from dromedaries. This finding expands the possible reservoirs of WNV and sources of WNV infection. PMID:27273223

  9. First isolation of West Nile virus from a dromedary camel.

    PubMed

    Joseph, Sunitha; Wernery, Ulrich; Teng, Jade Ll; Wernery, Renate; Huang, Yi; Patteril, Nissy Ag; Chan, Kwok-Hung; Elizabeth, Shyna K; Fan, Rachel Yy; Lau, Susanna Kp; Kinne, Jörg; Woo, Patrick Cy

    2016-01-01

    Although antibodies against West Nile virus (WNV) have been detected in the sera of dromedaries in the Middle East, North Africa and Spain, no WNV has been isolated or amplified from dromedary or Bactrian camels. In this study, WNV was isolated from Vero cells inoculated with both nasal swab and pooled trachea/lung samples from a dromedary calf in Dubai. Complete-genome sequencing and phylogenetic analysis using the near-whole-genome polyprotein revealed that the virus belonged to lineage 1a. There was no clustering of the present WNV with other WNVs isolated in other parts of the Middle East. Within lineage 1a, the dromedary WNV occupied a unique position, although it was most closely related to other WNVs of cluster 2. Comparative analysis revealed that the putative E protein encoded by the genome possessed the original WNV E protein glycosylation motif NYS at E154-156, which contained the N-linked glycosylation site at N-154 associated with increased WNV pathogenicity and neuroinvasiveness. In the putative NS1 protein, the A70S substitution observed in other cluster 2 WNVs and P250, which has been implicated in neuroinvasiveness, were present. In addition, the foo motif in the putative NS2A protein, which has been implicated in neuroinvasiveness, was detected. Notably, the amino-acid residues at 14 positions in the present dromedary WNV genome differed from those in most of the closely related WNV strains in cluster 2 of lineage 1a, with the majority of these differences observed in the putative E and NS5 proteins. The present study is the first to demonstrate the isolation of WNV from dromedaries. This finding expands the possible reservoirs of WNV and sources of WNV infection. PMID:27273223

  10. Isolation of Saint Louis Encephalitis Virus from a Horse with Neurological Disease in Brazil

    PubMed Central

    Rosa, Roberta; Costa, Erica Azevedo; Marques, Rafael Elias; Oliveira, Taismara Simas; Furtini, Ronaldo; Bomfim, Maria Rosa Quaresma; Teixeira, Mauro Martins; Paixão, Tatiane Alves; Santos, Renato Lima

    2013-01-01

    St. Louis encephalitis virus (SLEV) is a causative agent of encephalitis in humans in the Western hemisphere. SLEV is a positive-sense RNA virus that belongs to the Flavivirus genus, which includes West Nile encephalitis virus, Japanese encephalitis virus, Dengue virus and other medically important viruses. Recently, we isolated a SLEV strain from the brain of a horse with neurological signs in the countryside of Minas Gerais, Brazil. The SLEV isolation was confirmed by reverse-transcription RT-PCR and sequencing of the E protein gene. Virus identity was also confirmed by indirect immunofluorescence using commercial antibodies against SLEV. To characterize this newly isolated strain in vivo, serial passages in newborn mice were performed and led to hemorrhagic manifestations associated with recruitment of inflammatory cells into the central nervous system of newborns. In summary this is the first isolation of SLEV from a horse with neurological signs in Brazil. PMID:24278489

  11. Isolation and phylogenetic characterization of Canine distemper virus from India.

    PubMed

    Swati; Deka, Dipak; Uppal, Sanjeev Kumar; Verma, Ramneek

    2015-09-01

    Canine distemper (CD), caused by canine distemper virus (CDV) is a highly contagious disease that infects a variety of carnivores. Sequence analysis of CDVs from different geographical areas has shown a lot of variation in the genome of the virus especially in haemagglutinin gene which might be one of the causes of vaccine failure. In this study, we isolated the virus (place: Ludhiana, Punjab; year: 2014) and further cloned, sequenced and analyzed partial haemagglutinin (H) gene and full length genes for fusion protein (F), phosphoprotein (P) and matrix protein (M) from an Indian wild-type CDV. Higher sequence homology was observed with the strains from Switzerland, Hungary, Germany; and lower with the vaccine strains like Ondersteport, CDV3, Convac for all the genes. The multiple sequence alignment showed more variation in partial H (45 nucleotide and 5 amino acid substitutions) and complete F (79 nucleotide and 30 amino acid substitutions) than in complete P (44 nucleotide and 22 amino acid substitutions) and complete M (22 nucleotide and 4 amino acid substitutions) gene/protein. Predicted potential N-linked glycosylation sites in H, F, M and P proteins were similar to the previously known wild-type CDVs but different from the vaccine strains. The Indian CDV formed a distinct clade in the phylogenetic tree clearly separated from the previously known wild-type and vaccine strains. PMID:26396979

  12. Characterization of reticuloendotheliosis virus isolates obtained from chickens, turkeys and prairie chickens in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Twelve reticuloendotheliosis virus (REV) isolates obtained from chickens, turkeys and prairie chickens in the United States were characterized using ploymerase chain reaction (PCR) and indirect immunofluoresence (IFA) assays. This study included five REV isolates from Prairie chickens in Texas, two ...

  13. West Nile virus in Tunisia, 2014: First isolation from mosquitoes.

    PubMed

    Wasfi, F; Dachraoui, K; Cherni, S; Bosworth, A; Barhoumi, W; Dowall, S; Chelbi, I; Derbali, M; Zoghlami, Z; Beier, J C; Zhioua, E

    2016-07-01

    Several outbreaks of human West Nile virus (WNV) infections were reported in Tunisia during the last two decades. Serological studies on humans as well as on equine showed intensive circulation of WNV in Tunisia. However, no virus screening of mosquitoes for WNV has been performed in Tunisia. In the present study, we collected mosquito samples from Central Tunisia to be examined for the presence of flaviviruses. A total of 102 Culex pipiens mosquitoes were collected in September 2014 from Central Tunisia. Mosquitoes were pooled according to the collection site, date and sex with a maximum of 5 specimens per pool and tested for the presence of flaviviruses by conventional reverse transcription heminested PCR and by a specific West Nile virus real time reverse transcription PCR. Of a total of 21 pools tested, 7 were positive for WNV and no other flavivirus could be evidenced in mosquito pools. In addition, WNV was isolated on Vero cells. Phylogenetic analysis showed that recent Tunisian WNV strains belong to lineage 1 WNV and are closely related to the Tunisian strain 1997 (PAH 001). This is the first detection and isolation of WNV from mosquitoes in Tunisia. Some areas of Tunisia are at high risk for human WNV infections. WNV is likely to cause future sporadic and foreseeable outbreaks. Therefore, it is of major epidemiological importance to set up an entomological surveillance as an early alert system. Timely detection of WNV should prompt vector control to prevent future outbreaks. In addition, education of people to protect themselves from mosquito bites is of major epidemiological importance as preventive measure against WNV infection. PMID:27038557

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

    USGS Publications Warehouse

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

    2007-01-01

    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.

  15. Phylogenetic analysis of Bunyamwera and Ngari viruses (family Bunyaviridae, genus Orthobunyavirus) isolated in Kenya.

    PubMed

    Odhiambo, C; Venter, M; Lwande, O; Swanepoel, R; Sang, R

    2016-01-01

    Orthobunyaviruses, tri-segmented, negative-sense RNA viruses, have long been associated with mild to severe human disease in Africa, but not haemorrhagic fever. However, during a Rift Valley fever outbreak in East Africa in 1997-1998, Ngari virus was isolated from two patients and antibody detected in several others with haemorrhagic fever. The isolates were used to identify Ngari virus as a natural Orthobunyavirus reassortant. Despite their potential to reassort and cause severe human disease, characterization of orthobunyaviruses is hampered by paucity of genetic sequences. Our objective was to obtain complete gene sequences of two Bunyamwera virus and three Ngari virus isolates from recent surveys in Kenya and to determine their phylogenetic positioning within the Bunyamwera serogroup. Newly sequenced Kenyan Bunyamwera virus isolates clustered closest to a Bunyamwera virus isolate from the same locality and a Central African Republic isolate indicating that similar strains may be circulating regionally. Recent Kenyan Ngari isolates were closest to the Ngari isolates associated with the 1997-1998 haemorrhagic fever outbreak. We observed a temporal/geographical relationship among Ngari isolates in all three gene segments suggesting a geographical/temporal association with genetic diversity. These sequences in addition to earlier sequences can be used for future analyses of this neglected but potentially deadly group of viruses. PMID:26118981

  16. The equine arteritis virus isolate from the 2010 Argentinian outbreak.

    PubMed

    Metz, G E; Serena, M S; Panei, C J; Nosetto, E O; Echeverria, M G

    2014-12-01

    A semen sample from a stallion infected during the 2010 equine arteritis virus (EAV) outbreak was received for viral isolation prior to castration of the animal. The virus was identified using a polyclonal antibody immunofluorescence test. Reverse-transcription polymerase chain reaction (RT-PCR) was used to amplify a region of the GP5 gene with primers GL105F and GL673R. The PCR products were purified and sequences of both strands were determined in a MegaBACE™1000 with inner primers CR2 and EAV32. A phylogenetic dataset was built with the previously reported sequences of five strains isolated in Argentina, together with a group of selected sequences obtained from GenBank. The unrooted neighbour-joining tree was constructed using molecular evolutionary genetic analysis (MEGA) and bootstrap analyses were conducted using 1,000 replicate datasets. Evolutionary distances were computed using the maximum composite likelihood method. A NetNGlyc server analysis at the Technical University of Denmark (www.cbs.dtu.dk/services/NetNGlyc/) was used to predict N-glycosylation in GP5 sequences. The phylogenetic analysis revealed that the new strain GLD-LP-ARG), together with other strains previously isolated, belongs to the European group EU-1 but in a different branch. The new strain shows 99% nucleotide identity with strain Al1and 98.1% with the Belgian strain 08P178. Persistently infected stallions and their cryopreserved semen constitute a reservoir of EAV, which ensures its persistence in the horse population around the world. These findings reinforce the importance of careful monitoring of persistently infected stallions, as well as semen straws, by RT-PCR or test mating, in accordance with national regulations. PMID:25812217

  17. Isolation of sochi virus from a fatal case of hantavirus disease with fulminant clinical course.

    PubMed

    Dzagurova, Tamara K; Witkowski, Peter T; Tkachenko, Evgeniy A; Klempa, Boris; Morozov, Vyacheslav G; Auste, Brita; Zavora, Dmitriy L; Iunicheva, Iulia V; Mutnih, Elena S; Kruger, Detlev H

    2012-01-01

    Sochi virus, a novel genetic variant of Dobrava-Belgrade virus, was isolated in cell culture from a fulminant lethal case of hantavirus disease presenting with shock and combined kidney and lung failure. Sochi virus is transmitted to humans from host reservoir Apodemus ponticus and must be considered a life-threatening emerging agent. PMID:22042875

  18. Complete Genome Sequence of a Street Rabies Virus Isolated from a Dog in Nigeria

    PubMed Central

    Zhou, Ming; Zhou, Zutao; Kia, Grace S. N.; Gnanadurai, Clement W.; Leyson, Christina M.; Umoh, Jarlath U.; Kwaga, Jacob P.; Kazeem, Haruna M.

    2013-01-01

    A canine rabies virus (RABV) was isolated from a trade dog in Nigeria. Its entire genome was sequenced and found to be closely related to canine RABVs circulating in Africa. Sequence comparison indicates that the virus is closely related to the Africa 2 RABV lineage. The virus is now termed DRV-NG11. PMID:23469344

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

    PubMed Central

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

    2014-01-01

    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

  20. Genetic relationships between southern African SAT-2 isolates of foot-and-mouth-disease virus.

    PubMed Central

    Vosloo, W.; Knowles, N. J.; Thomson, G. R.

    1992-01-01

    Sequencing of part of the 1D gene of foot-and-mouth disease virus was used to determine the relationships between SAT-2 viruses isolated from outbreaks which occurred in cattle in Zimbabwe and Namibia and in impala in South Africa between 1979 and 1989. The results demonstrated that the outbreaks in different countries were unrelated. Surprisingly close relationships were shown between all SAT-2 viruses isolated from cattle in Zimbabwe since 1983 but the two major epizootics which occurred in 1989 were caused by viruses which were clearly different. Conversely, two apparently unrelated outbreaks in impala in South Africa were caused by viruses which could not be distinguished. PMID:1334842

  1. Acquistion and transmissibility of United States Soybean dwarf virus isolates by the soybean aphid Aphis glycines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Soybean dwarf virus (SbDV) exists as several distinct strains based on symptomatology, vectors, and host range. Original Japanese isolates of SbDV were specifically transmitted by Aulacorthum solani. More recently Japanese isolates, other exotic isolates, and endemic U.S. isolates have been shown ...

  2. Isolation of caprine arthritis encephalitis virus from goats in Mexico.

    PubMed Central

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

    1999-01-01

    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

  3. Complete Genome Sequence of Papaya Ringspot Virus Isolated from Genetically Modified Papaya in Hainan Island, China.

    PubMed

    Zhao, Guangyuan; Yan, Pu; Shen, Wentao; Tuo, Decai; Li, Xiaoying; Zhou, Peng

    2015-01-01

    The complete genome sequence (10,326 nucleotides) of a papaya ringspot virus isolate infecting genetically modified papaya in Hainan Island of China was determined through reverse transcription (RT)-PCR. The virus shares 92% nucleotide sequence identity with the isolate that is unable to infect PRSV-resistant transgenic papaya. PMID:26358610

  4. Genetic characterization of epizootic hemorrhagic disease virus strains isolated from cattle in Israel

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Epizootic hemorrhagic disease virus (EHDV), an Orbivirus not previously reported in Israel, was isolated from Israeli cattle during a “bluetongue like” disease outbreak in 2006. To ascertain the origin of this new virus, three isolates from the outbreak were fully sequenced and compared with availab...

  5. Phylogenetic analysis of Newcastle disease virus isolates occurring in India during 1989-2013.

    PubMed

    Desingu, P A; Singh, S D; Dhama, K; Karthik, K; Vinodh Kumar, O R; Malik, Y S

    2016-06-01

    The study details characterization of Newcastle disease virus (NDV) isolates recovered from commercial poultry flocks (chicken) and wild birds (crane) of India during the time period from 1989 to 2013. Phylogenetic analysis revealed that most of the NDV isolates belongs to class II, genotype XIIIa and a chicken isolate (108/BAREILLY/AD-IVRI/91) was of genotype VI, where it showed diversity of 3 % from the other viruses belonging to same genotype. Another chicken isolate (75/RAMPUR/AD-IVRI/89) grouped in genotype III and showed 4 % diversity with viruses of genotype III. The crane origin NDV identified as of genotype II corresponding to the vaccine virus. This appears to be the first report about existence of genotype XIIIa and its ancestral viruses are circulating in India for the last two decades in different species of birds. Furthermore, genetically distinct viruses belonging to genotypes II, III and VI are also circulating in India. PMID:27366774

  6. First Complete Genome Sequence of a Chikungunya Virus Strain Isolated from a Patient Diagnosed with Dengue Virus Infection in Malaysia

    PubMed Central

    Gan, Han Ming; Rohani, Ahmad

    2016-01-01

    Here, we report the complete genome sequence of a chikungunya virus coinfection strain isolated from a dengue virus serotype 2-infected patient in Malaysia. This coinfection strain was determined to be of the Asian genotype and contains a novel insertion in the nsP3 gene. PMID:27563048

  7. First Complete Genome Sequence of a Chikungunya Virus Strain Isolated from a Patient Diagnosed with Dengue Virus Infection in Malaysia.

    PubMed

    Ooi, Man Kwan; Gan, Han Ming; Rohani, Ahmad; Syed Hassan, Sharifah

    2016-01-01

    Here, we report the complete genome sequence of a chikungunya virus coinfection strain isolated from a dengue virus serotype 2-infected patient in Malaysia. This coinfection strain was determined to be of the Asian genotype and contains a novel insertion in the nsP3 gene. PMID:27563048

  8. Atypical myxomatosis--virus isolation, experimental infection of rabbits and restriction endonuclease analysis of the isolate.

    PubMed

    Psikal, I; Smíd, B; Rodák, L; Valícek, L; Bendová, J

    2003-08-01

    Atypical form of myxomatosis, which caused non-lethal and clinically mild disease in domestic rabbits 1 month after immunization with a commercially available vaccine MXT, is described. The isolated myxoma virus designated as Litovel 2 (Li-2) did not induce systemic disease following subcutaneous and intradermal applications in susceptible experimental rabbits but led to the immune response demonstrated by ELISA. No severe disease was induced in those Li-2 inoculated rabbits by challenge with the virulent strains Lausanne (Lu) or Sanar (SA), while the control animals showed nodular form of myxomatosis with lethal course of the illness. Restriction fragment length polymorphism (RFLP) of genomic DNA with KpnI and BamHI endonucleases was used for genetic characterization of the Li-2 isolate, the vaccine strain MXT and both virulent strains Lu and SA, respectively. In general, RFLP analysis has shown to be informative for inferring genetic relatedness between myxoma viruses. Based on restriction endonuclease DNA fragment size distribution, it was evident that the pathogenic strain SA is genetically related to the reference strain Lu and the isolate Li-2 is more related, but not identical, to the vaccination strain MXT. PMID:14628995

  9. Severe acute measles pneumonitis: virus isolation in bronchoalveolar lavage fluid.

    PubMed

    Suter, Cosima; Buergi, Urs; Eigenmann, Katja; Franzen, Daniel

    2015-01-01

    In the past few years, several endemic outbreaks of measles have been recognised, not only in children but also in adults, with severe and, occasionally, even fatal complications, possibly due to delayed diagnosis of the disease in adult medicine and decreasing vaccination rates. Furthermore, the treatment consists of supportive measures only. We present a case of severe measles pneumonitis in a 42-year-old man, a travel returnee, proven by direct virus isolation with PCR from bronchoalveolar lavage fluid. CT findings and pulmonary function testing revealed features of obstructive bronchiolitis; the patient was successfully treated with corticosteroids. He fully recovered, and lung function measurement returned to normal values. We conclude that acute measles can present as obstructive bronchiolitis and may be successfully treated with corticosteroids. PMID:26508116

  10. Detection, isolation, and persistence of viruses within bivalve mollusks

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Norovirus (NV), hepatitis A virus (HAV), and other virus transmission by molluscan shellfish is a significant issue. Research at the ARS-Dover DE laboratory has led to the development of improved methods for detecting these viruses. To identify pathogenic viruses within mollusks, a rapid highly-se...

  11. Genetic characterization of rabies viruses isolated from frugivorous bat (Artibeus spp.) in Brazil.

    PubMed

    Shoji, Youko; Kobayashi, Yuki; Sato, Go; Itou, Takuya; Miura, Yasuo; Mikami, Takeshi; Cunha, Elenice M S; Samara, Samir I; Carvalho, Adlorata A B; Nocitti, Darci P; Ito, Fumio H; Kurane, Ichiro; Sakai, Takeo

    2004-10-01

    In Latin America, rabies cases related to frugivorous bats have been reported since 1930's. Recently, two viruses isolated from Artibeus lituratus were proved to be vampire bat variants by monoclonal antibodies panels [2], but their genetic information is not well known. In this report, four rabies viruses were isolated from frugivorous bats (Artibeus spp.) in Brazil and their nucleoprotein gene sequences were determined. These isolates were found to be genotype 1 of lyssavirus and showed the maximum nucleotide sequence homology of 97.6-99.4% with vampire bat-related viruses in Brazil [6]. These results indicate that the Brazilian frugivorous bat rabies viruses in this study are closely related to vampire bat-related viruses that play a main role in rabies virus transmission to livestock in Brazil. PMID:15528863

  12. Isolation of new Brazilian giant viruses from environmental samples using a panel of protozoa

    PubMed Central

    Dornas, Fábio P.; Khalil, Jacques Y. B.; Pagnier, Isabelle; Raoult, Didier; Abrahão, Jônatas; La Scola, Bernard

    2015-01-01

    The Megavirales are a newly described order capable of infecting different types of eukaryotic hosts. For the most part, the natural host is unknown. Several methods have been used to detect these viruses, with large discrepancies between molecular methods and co-cultures. To isolate giant viruses, we propose the use of different species of amoeba as a cellular support. The aim of this work was to isolate new Brazilian giant viruses by comparing the protozoa Acanthamoeba castellanii, A. polyphaga, A. griffini, and Vermamoeba vermiformis (VV) as a platform for cellular isolation using environmental samples. One hundred samples were collected from 3 different areas in September 2014 in the Pampulha lagoon of Belo Horizonte city, Minas Gerais, Brazil. PCR was used to identify the isolated viruses, along with hemacolor staining, labelling fluorescence and electron microscopy. A total of 69 viruses were isolated. The highest ratio of isolation was found in A. polyphaga (46.38%) and the lowest in VV (0%). Mimiviruses were the most frequently isolated. One Marseillevirus and one Pandoravirus were also isolated. With Brazilian environmental samples, we demonstrated the high rate of lineage A mimiviruses. This work demonstrates how these viruses survive and circulate in nature as well the differences between protozoa as a platform for cellular isolation. PMID:26500630

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

    SciTech Connect

    Frenkel, N.; Schirmer, E.C.; Wyatt, L.S.; Katsafanas, G.; Roffman, E.; Danovich, R.M. ); June, C.H. )

    1990-01-01

    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.

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

    SciTech Connect

    Foley, Brian T; Korber, Bette T

    2008-01-01

    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.

  15. Phylogenetic analysis of Newcastle disease viruses isolated from commercial poultry in Mozambique (2011-2016).

    PubMed

    Mapaco, Lourenço P; Monjane, Iolanda V A; Nhamusso, Antonieta E; Viljoen, Gerrit J; Dundon, William G; Achá, Sara J

    2016-10-01

    The complete sequence of the fusion (F) protein gene from 11 Newcastle disease viruses (NDVs) isolated from commercial poultry in Mozambique between 2011 and 2016 has been generated. The F gene cleavage site motif for all 11 isolates was (112)RRRKRF(117) indicating that the viruses are virulent. A phylogenetic analysis using the full F gene sequence revealed that the viruses clustered within genotype VIIh and showed a higher similarity to NDVs from South Africa, China and Southeast Asia than to viruses previously described in Mozambique in 1994, 1995 and 2005. The identification of these new NDVs has important implications for Newcastle disease management and control in Mozambique. PMID:27277578

  16. Avian influenza virus isolation, propagation and titration in embryonated chicken eggs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Avian influenza (AI) virus is usually isolated, propagated, and titrated in embryonated chickens eggs (ECE). Most any sample type can be accommodated for culture with appropriate processing. Isolation may also be accomplished in cell culture particularly if mammalian lineage isolates are suspected, ...

  17. PATHOGENESIS OF CHICKEN-PASSAGED NEWCASTLE DISEASE VIRUSES ISOLATED FROM CHICKENS, WILD, AND EXOTIC BIRDS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The pathogenesis of six Newcastle disease virus (NDV) isolates recovered from chickens and wild (anhinga) and exotic (yellow nape parrot, pheasant, and dove isolate) birds was examined after four passages of the isolates in domestic chickens. Groups of four-week-old specific-pathogen-free White Legh...

  18. Recombinant virus assay: a rapid, phenotypic assay for assessment of drug susceptibility of human immunodeficiency virus type 1 isolates.

    PubMed Central

    Kellam, P; Larder, B A

    1994-01-01

    Antiviral drug susceptibility assays for clinical human immunodeficiency virus type 1 (HIV-1) isolates are required to monitor the development of drug resistance during clinical trials and antiretroviral drug therapy. First-generation phenotypic assays possess a number of drawbacks, not least the selection of unrepresentative virus populations during cocultivation. Here we describe a rapid phenotypic assay for the assessment of the susceptibility of clinical isolates to reverse transcriptase (RT) inhibitors. This procedure, called the recombinant virus assay, allows the generation of viable virus by homologous recombination of a PCR-derived pool of RT coding sequences into an RT-deleted, noninfectious proviral clone, pHIV delta BstEII. A nested PCR procedure has been optimized to allow the amplification of an RT pool from both uncultured and cocultured infected patient peripheral blood lymphocyte (PBL) DNA for subsequent use in the creation of recombinant viruses. Analysis of two patients during the course of zidovudine therapy showed that this approach produced viruses which accurately exhibited the same genotype and phenotype as that of the original infected PBL DNA. The recombinant virus assay can be performed in approximately 3 weeks without the use of donor PBLs and therefore represents a rapid, nonselective procedure for the assay of clinical isolates. Images PMID:8141575

  19. Complete Genome Sequence of Goose Tembusu Virus, Isolated from Jiangnan White Geese in Jiangsu, China

    PubMed Central

    Han, Kaikai; Huang, Xinmei; Zhao, Dongmin; Liu, Yuzhuo; Zhou, Xiaobo; You, Yuan; Xie, Xingxing

    2013-01-01

    Avian tembusu virus (TMUV), which was first identified in eastern China, is an emerging virus causing serious economic losses in the Chinese poultry industry. Here, we report the complete genome sequence of goose tembusu virus strain JS804, isolated from Jiangnan white geese with severe neurological signs. The genome of JS804 is 10,990 nucleotides (nt) in length and contains a single open reading frame encoding a putative polyprotein of 3,425 amino acids. Research of the whole sequence of tembusu virus will help us to understand further the molecular and evolutionary characteristics and pathogenesis of this virus. PMID:23516233

  20. Isolation of Tacaribe Virus, a Caribbean Arenavirus, from Host-Seeking Amblyomma americanum Ticks in Florida

    PubMed Central

    Sayler, Katherine A.; Barbet, Anthony F.; Chamberlain, Casey; Clapp, William L.; Alleman, Rick; Loeb, Julia C.; Lednicky, John A.

    2014-01-01

    Arenaviridae are a family of single stranded RNA viruses of mammals and boid snakes. Twenty-nine distinct mammalian arenaviruses have been identified, many of which cause severe hemorrhagic disease in humans, particularly in parts of sub-Saharan Africa, and in Central and South America. Humans typically become infected with an arenavirus through contact with excreta from infected rodents. Tacaribe virus (TCRV) is an arenavirus that was first isolated from bats and mosquitoes during a rabies surveillance survey conducted in Trinidad from 1956 to 1958. Tacaribe virus is unusual because it has never been associated with a rodent host and since that one time isolation, the virus has not been isolated from any vertebrate or invertebrate hosts. We report the re-isolation of the virus from a pool of 100 host-seeking Amblyomma americanum (lone star ticks) collected in a Florida state park in 2012. TCRV was isolated in two cell lines and its complete genome was sequenced. The tick-derived isolate is nearly identical to the only remaining isolate from Trinidad (TRVL-11573), with 99.6% nucleotide identity across the genome. A quantitative RT-PCR assay was developed to test for viral RNA in host-seeking ticks collected from 3 Florida state parks. Virus RNA was detected in 56/500 (11.2%) of surveyed ticks. As this virus was isolated from ticks that parasitize humans, the ability of the tick to transmit the virus to people should be evaluated. Furthermore, reservoir hosts for the virus need to be identified in order to develop risk assessment models of human infection. PMID:25536075

  1. VIRULENCE OF HETEROGENEOUS-ORIGIN NEWCASTLE DISEASE VIRUS ISOLATES BEFORE AND AFTER SEQUENTIAL PASSAGES IN DOMESTIC CHICKENS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Four serial passages of six Newcastle disease virus (NDV) isolates were performed in 2-week-old White Leghorns. The viruses were recovered from chickens (Ckn-Live Bird Market and Ckn-Australia isolates), exotic (Yellow Nape [YN] Parrot, Pheasant, and Dove isolates) and wild birds (Anhinga isolate). ...

  2. Detection and isolation of Bluetongue virus from commercial vaccine batches.

    PubMed

    Bumbarov, Velizar; Golender, Natalia; Erster, Oran; Khinich, Yevgeny

    2016-06-14

    In this report we describe the detection and identification of Bluetongue virus (BTV) contaminations in commercial vaccines. BTV RNA was detected in vaccine batches of Lumpy skin disease (LSD) and Sheep pox (SP) using quantitative PCR (qPCR) for VP1 and NS3 genes. Both batches were positive for VP1 and NS3 in qPCR. The LSD vaccine-derived sample was positive for VP1 and VP2 in conventional PCR. The SP vaccine-derived sample was examined by amplification of VP1, VP4, VP6, VP7, NS2 and NS3 gene segments in conventional PCR. The SP vaccine-derived sample was further propagated in embryonated chicken eggs (ECE) and Vero cells. Preliminary sequence analysis showed that the LSD vaccine-derived sequence was 98-99% similar to BTV9. Analysis of the six genomic segments from the SP vaccine-derived isolate showed the highest similarity to BTV26 (66.3-97.8%). These findings are particularly important due to the effect of BTV on cattle and sheep, for which the vaccines are intended. They also demonstrate the necessity of rigorous vaccine inspection and strict vaccine production control. PMID:27171751

  3. Dengue Virus Envelope Dimer Epitope Monoclonal Antibodies Isolated from Dengue Patients Are Protective against Zika Virus

    PubMed Central

    Swanstrom, J. A.; Plante, J. A.; Plante, K. S.; Young, E. F.; McGowan, E.; Gallichotte, E. N.; Widman, D. G.; Heise, M. T.; de Silva, A. M.

    2016-01-01

    ABSTRACT Zika virus (ZIKV) is a mosquito-borne flavivirus responsible for thousands of cases of severe fetal malformations and neurological disease since its introduction to Brazil in 2013. Antibodies to flaviviruses can be protective, resulting in lifelong immunity to reinfection by homologous virus. However, cross-reactive antibodies can complicate flavivirus diagnostics and promote more severe disease, as noted after serial dengue virus (DENV) infections. The endemic circulation of DENV in South America and elsewhere raises concerns that preexisting flavivirus immunity may modulate ZIKV disease and transmission potential. Here, we report on the ability of human monoclonal antibodies and immune sera derived from dengue patients to neutralize contemporary epidemic ZIKV strains. We demonstrate that a class of human monoclonal antibodies isolated from DENV patients neutralizes ZIKV in cell culture and is protective in a lethal murine model. We also tested a large panel of convalescent-phase immune sera from humans exposed to primary and repeat DENV infection. Although ZIKV is most closely related to DENV compared to other human-pathogenic flaviviruses, most DENV immune sera (73%) failed to neutralize ZIKV, while others had low (50% effective concentration [EC50], <1:100 serum dilution; 18%) or moderate to high (EC50, >1:100 serum dilution; 9%) levels of cross-neutralizing antibodies. Our results establish that ZIKV and DENV share epitopes that are targeted by neutralizing, protective human antibodies. The availability of potently neutralizing human monoclonal antibodies provides an immunotherapeutic approach to control life-threatening ZIKV infection and also points to the possibility of repurposing DENV vaccines to induce cross-protective immunity to ZIKV. PMID:27435464

  4. Neurological lesions in chickens experimentally infected with virulent Newcastle disease virus isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Neuropil reaction was evaluated in chickens inoculated with four different Newcastle disease virus (NDV) isolates, including Texas GB, Turkey North Dakota, Nevada Cormorant (velogenic neurotropic) and Anhinga (mesogenic). Tissues for this study included archived formalin-fixed, paraffin embedded br...

  5. Use of muscovy duck embryo fibroblasts for the isolation of viruses from wild birds

    USGS Publications Warehouse

    Docherty, D.E.; Slota, Paul G.

    1988-01-01

    Techniques are described for the preparation, cryopreservation, and inoculation of Muscovy duck embryo cell cultures. The procedure yields a susceptible reproducible cell culture system for the isolation and cultivation of viruses from wild birds.

  6. Complete Genome Sequence of a Dengue Virus Serotype 4 Strain Isolated in Roraima, Brazil

    PubMed Central

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

    2012-01-01

    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

  7. Complete Genome Sequence of Reticuloendotheliosis Virus Strain MD-2, Isolated from a Contaminated Turkey Herpesvirus Vaccine

    PubMed Central

    Li, Junping; Yang, Chenghuai; Li, Qihong; Li, Huijiao; Xia, Yecai; Liu, Dan

    2013-01-01

    Here, we present the complete genomic sequence of a reticuloendotheliosis virus (REV) isolated from a contaminated turkey herpesvirus (HVT) vaccine. This report will be helpful for epidemiological studies on REV infection in avian flocks. PMID:24092783

  8. Complete Genome Sequence of Viral Hemorrhagic Septicemia Virus Isolated from an Olive Flounder in South Korea

    PubMed Central

    Kim, Jong-Oh; Kim, Wi-Sik; Nishizawa, Toyohiko

    2013-01-01

    Viral hemorrhagic septicemia virus (VHSV) is a seriously problematic pathogen in olive flounder (Paralichthys olivaceus) aquaculture farms in South Korea. Here, we report the complete genome sequence of VHSV which was isolated from spleen and kidney tissues of dead fish at an aquaculture farm in 2005. This genome sequence will be useful for virus diagnostics and in comparative analyses with other virus genotypes. PMID:24009117

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

    PubMed

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

    2009-01-01

    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

  10. West Nile virus isolated from a Virginia opossum (Didelphis virginiana) in northwestern Missouri, USA, 2012.

    PubMed

    Bosco-Lauth, Angela; Harmon, Jessica R; Lash, R Ryan; Weiss, Sonja; Langevin, Stanley; Savage, Harry M; Godsey, Marvin S; Burkhalter, Kristen; Root, J Jeffrey; Gidlewski, Thomas; Nicholson, William L; Brault, Aaron C; Komar, Nicholas

    2014-10-01

    We describe the isolation of West Nile virus (WNV; Flaviviridae, Flavivirus) from blood of a Virginia opossum (Didelphis virginiana) collected in northwestern Missouri, USA in August 2012. Sequencing determined that the virus was related to lineage 1a WNV02 strains. We discuss the role of wildlife in WNV disease epidemiology. PMID:25098303

  11. West Nile Virus Isolated from a Virginia Opossum (Didelphis virginiana) in Northwestern Missouri, USA, 2012

    PubMed Central

    Bosco-Lauth, Angela; Harmon, Jessica R.; Lash, R. Ryan; Weiss, Sonja; Langevin, Stanley; Savage, Harry M.; Godsey, Marvin S.; Burkhalter, Kristen; Root, J. Jeffrey; Gidlewski, Thomas; Nicholson, William L.; Brault, Aaron C.; Komar, Nicholas

    2016-01-01

    We describe the isolation of West Nile virus (WNV; Flaviviridae, Flavivirus) from blood of a Virginia opossum (Didelphis virginiana) collected in northwestern Missouri, USA in August 2012. Sequencing determined that the virus was related to lineage 1a WNV02 strains. We discuss the role of wildlife in WNV disease epidemiology. PMID:25098303

  12. Full-Genome Sequence of a Novel Varicella-Zoster Virus Clade Isolated in Mexico

    PubMed Central

    Rodríguez-Castillo, Araceli; Ortiz-Alcántara, Joanna María; Gonzalez-Durán, Elizabeth; Segura-Candelas, José Miguel; Pérez-Agüeros, Sandra Ivette; Escobar-Escamilla, Noé; Méndez-Tenorio, Alfonso; Diaz-Quiñonez, José Alberto

    2015-01-01

    Varicella-zoster virus (VZV) is a member of the Herpesviridae family, which causes varicella (chicken pox) and herpes zoster (shingles) in humans. Here, we report the complete genome sequence of varicella-zoster virus, isolated from a vesicular fluid sample, revealing the circulation of VZV clade VIII in Mexico. PMID:26159533

  13. Full-Genome Sequence of a Novel Varicella-Zoster Virus Clade Isolated in Mexico.

    PubMed

    Garcés-Ayala, Fabiola; Rodríguez-Castillo, Araceli; Ortiz-Alcántara, Joanna María; Gonzalez-Durán, Elizabeth; Segura-Candelas, José Miguel; Pérez-Agüeros, Sandra Ivette; Escobar-Escamilla, Noé; Méndez-Tenorio, Alfonso; Diaz-Quiñonez, José Alberto; Ramirez-González, José Ernesto

    2015-01-01

    Varicella-zoster virus (VZV) is a member of the Herpesviridae family, which causes varicella (chicken pox) and herpes zoster (shingles) in humans. Here, we report the complete genome sequence of varicella-zoster virus, isolated from a vesicular fluid sample, revealing the circulation of VZV clade VIII in Mexico. PMID:26159533

  14. Complete genome sequence of Tomato mosaic virus isolated from jasmine in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tomato mosaic virus (ToMV) was first identified in jasmine in the U.S. in Florida in 1999. This report provides the first full genome sequence of a ToMV isolate from jasmine. The full genome sequence of this virus will enable research scientists to develop additional specific diagnostic tests for ...

  15. Complete genome sequence of a Tomato mottle mosaic virus isolate from the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tomato mottle mosaic virus (ToMMV) was first identified in the U.S. in tomatoes in Florida in 2010. This report provides the first full genome sequence of a U.S. ToMMV isolate from 2010. The full genome sequence of this emerging virus will enable research scientists to develop additional specific ...

  16. Characterization of the Taura syndrome virus isolate originating from the 2004 Texas Epizootic in cultured shrimp

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Taura syndrome virus (TSV) is a major viral pathogen of penaeid shrimp worldwide. A comprehensive investigation of the Texas isolate of TSV that caused epizootics in shrimp farms in Texas in 2004 (Us04Pv1) revealed that the virus was highly virulent in laboratory bioassays causing severe symptom dev...

  17. Squash vein yellowing virus, a novel ipomovirus, isolated from squash and watermelon in Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A novel whitefly-transmitted member of the family Potyviridae was isolated from a squash plant (Cucurbita pepo) with vein yellowing symptoms in Florida. The virus, for which the name Squash vein yellowing virus (SqVYV) is proposed, has flexuous rod-shaped particles of ~840 nm in length. SqVYV was ...

  18. Characterization of low pathogenicity avian influenza viruses isolated from wild birds in Mongolia 2005 through 2007

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During 2005, 2006 and 2007 2,139 specimens representing 4,077 individual birds of 45 species were tested for avian influenza virus (AIV) as part of a wild bird AIV monitoring program conducted in Mongolia. Samples collected in 2005 were tested by virus isolation directly, samples from 2006 and 2007...

  19. West Nile virus isolated from Virginia opossum (Didelphis virginiana) in Northwest Missouri 2012

    DOE PAGESBeta

    Bosco-Lauth, Angela; Harmon, Jessica; Lash, R. Ryan; Weiss, Sonja; Langevin, Stanley; Savage, Harry; Marvin S. Godsey, Jr.; Burkhalter, Kristen; Root, J. Jeffrey; Gidlewski, Thomas; et al

    2014-12-01

    We describe the isolation of West Nile virus (WNV; Flaviviridae, flavivirus) from blood of a Virginia opossum (Didelphis virginiana) collected in northwestern Missouri, USA in August 2012. Furthermore, sequencing determined that the virus was related to lineage 1a WNV02 strains. We discuss the role of wildlife in WNV disease epidemiology.

  20. Complete Coding Sequences of Six Toscana Virus Strains Isolated from Human Patients in France

    PubMed Central

    Leparc-Goffart, Isabelle; Piorkowski, Geraldine; Coutard, Bruno; Papageorgiou, Nicolas; De Lamballerie, Xavier

    2016-01-01

    Toscana virus (TOSV) is an arthropod-borne phlebovirus belonging to the Sandfly fever Naples virus species (genus Phlebovirus, family Bunyaviridae). Here, we report the complete coding sequences of six TOSV strains isolated from human patients having acquired the infection in southeastern France during a 12-year period. PMID:27231377

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An outbreak of virulent Newcastle Disease Virus (NDV) in wild double-breasted cormorants (Phalacrocorax auritus) occurred in North America in the summer of 2008. All ten viruses isolated from cormorants were positively identified by the USDA validated real-time reverse transcriptase polymerase chai...

  2. Studies on antigenic and genomic properties of Brazilian rabies virus isolates.

    PubMed

    Schaefer, R; Batista, H B R; Franco, A C; Rijsewijk, F A M; Roehe, P M

    2005-05-20

    Despite the recognized stability of rabies virus, differences among isolates from different species have been found. This work was carried out with the aim to identify antigenic and genomic differences in Brazilian rabies virus isolates and to verify whether such alterations would bear any relationship with the different hosts for the virus in nature. For that, 79 Brazilian rabies viruses isolated from different host species and from distinct regions within Brazil were submitted to antigenic characterization with a panel of 11 monoclonal antibodies (Mabs) directed to lyssavirus antigens and to genomic analyses by the reverse transcriptase-polymerase chain reaction (RT-PCR) amplification of the N gene followed by restriction endonuclease analysis (REA). In addition, the nucleotide sequences of part of the N gene (225 bp) of seven isolates, taken as representative of the majority of the viruses under study, were determined. The analyses with the Mabs and RT-PCR/REA allowed the identification of two major groups of variants, the first formed by most isolates of cattle and bats and the second formed by viruses of dog origin. Partial sequencing of the N gene confirmed the similarity among isolates from cattle origin and those of vampire bats. However, viruses from non-haematophagous bats exhibited consistent differences from those of vampire bat isolates. Such findings suggest that the variants have evolved fairly stable modifications, which are not altered after passage in a dead-end host of a distinct species. No association could be established between antigenic or genomic alterations and geographic distribution of the isolates, which suggests that evolution of the virus has been directed to adaptation to the host species. PMID:15863275

  3. Complete Genome Sequences of Two Waterfowl-Origin Tembusu Virus Strains Isolated in Shandong Province, China

    PubMed Central

    Chen, Hao; Liu, Xin; Tang, Yi; Zhang, Ying; Ti, Jinfeng; Gao, Xuhui

    2013-01-01

    Here, we report the complete genome sequences of two tembusu virus strains, ZC-1 and LQ-1, isolated from ducks and geese, respectively, in 2012. Phylogenetic analysis showed that the nucleotide and amino acid sequences of the two strains are closely related to those of the TMUV isolates around Shandong province. The full-length genome sequences of two waterfowl-origin TMUVs provided herein will help to understand the molecular epidemiology of tembusu virus in China, which deserves further investigation. PMID:24356821

  4. Comparison of biological characteristics of H9N2 avian influenza viruses isolated from different hosts.

    PubMed

    Zhu, Yinbiao; Yang, Yang; Liu, Wei; Liu, Xin; Yang, Da; Sun, Zhihao; Ju, Yong; Chen, Sujuan; Peng, Daxin; Liu, Xiufan

    2015-04-01

    The pathogenicity and transmissibility of H9N2 influenza viruses has been widely investigated; however, few studies comparing the biological characteristics of H9N2 viruses isolated from different hosts have been performed. In this study, eight H9N2 viruses, isolated from chickens (Ck/F98, Ck/AH and Ck/TX), pigeons (Pg/XZ), quail/(Ql/A39), ducks (Dk/Y33) and swine (Sw/YZ and Sw/TZ) were selected, and their biological characteristics were determined. The results showed that all H9N2 viruses maintained a preference for both the avian- and human-type receptors, except for Sw/TZ, which had exclusive preference for the human-type receptor. The viruses replicated well in DF-1 and MDCK cells, whereas only three isolates, Ck/F98, Ck/TX and Sw/TZ, could replicate in A549 cells and also replicated in mouse lungs, resulting in body weight loss in mice. All H9N2 viruses were nonpathogenic to chickens and were detected in the trachea and lung tissues. The viruses were shed primarily by the oropharynx and were transmitted efficiently to naïve contact chickens. Our findings suggest that all H9N2 viruses from different hosts exhibit efficient replication and contact-transmission among chickens, and chickens serve as a good reservoir for the persistence and interspecies transmission of H9N2 influenza viruses. PMID:25616845

  5. Detection and Isolation of H5N1 Influenza virus from Large Volumes of Natural Water

    PubMed Central

    Khalenkov, Alexey; Laver, W. Graeme; Webster, Robert G.

    2009-01-01

    Various species of aquatic or wetlands birds can be the natural reservoir of avian influenza A viruses of all hemagglutinin (HA) subtypes. Shedding of the virus into water leads to transmission between waterfowl and is a major threat for epidemics in poultry and pandemics in humans. Concentrations of the influenza virus in natural water reservoirs are often too low to be detected by most methods. The procedure was designed to detect low concentrations of the influenza virus in large volumes of water without the need for costly installations and reagents. The virus was adsorbed onto formalin-fixed erythrocytes and subsequently isolated in chicken embryos. Sensitivity of the method was determined using a reverse-genetic H5N1 virus. A concentration as low as 0.03 of the 50% egg infection dose per milliliter (EID50/ml) of the initial volume of water was effectively detected. The probability of detection was ∼13%, which is comparable to that of detecting the influenza virus M-gene by PCR amplification. The method can be used by field workers, ecologists, ornithologists, and researchers who need a simple method to isolate H5N1 influenza virus from natural reservoirs. The detection and isolation of virus in embryonated chicken eggs may help epidemiologic, genetic, and vaccine studies. PMID:18325605

  6. Isolation of Madre de Dios Virus (Orthobunyavirus; Bunyaviridae), an Oropouche Virus Species Reassortant, from a Monkey in Venezuela.

    PubMed

    Navarro, Juan-Carlos; Giambalvo, Dileyvic; Hernandez, Rosa; Auguste, Albert J; Tesh, Robert B; Weaver, Scott C; Montañez, Humberto; Liria, Jonathan; Lima, Anderson; Travassos da Rosa, Jorge Fernando Soares; da Silva, Sandro P; Vasconcelos, Janaina M; Oliveira, Rodrigo; Vianez, João L S G; Nunes, Marcio R T

    2016-08-01

    Oropouche virus (OROV), genus Orthobunyavirus, family Bunyaviridae, is an important cause of human illness in tropical South America. Herein, we report the isolation, complete genome sequence, genetic characterization, and phylogenetic analysis of an OROV species reassortant, Madre de Dios virus (MDDV), obtained from a sick monkey (Cebus olivaceus Schomburgk) collected in a forest near Atapirire, a small rural village located in Anzoategui State, Venezuela. MDDV is one of a growing number of naturally occurring OROV species reassortants isolated in South America and was known previously only from southern Peru. PMID:27215299

  7. Genetic characterization and evolutionary analysis of Newcastle disease virus isolated from domestic duck in South Korea.

    PubMed

    Gaikwad, Satish; Kim, Ji-Ye; Lee, Hyun-Jeong; Jung, Suk Chan; Choi, Kang-Seuk

    2016-03-15

    Domestic ducks are considered a potential reservoir of Newcastle disease virus. In the study, a Newcastle disease virus (NDV) isolated from a domestic duck during surveillance in South Korea was characterized. The complete genome of the NDV isolate was sequenced, and the phylogenetic relationship to reference strains was studied. Phylogenetic analysis revealed that the strain clustered in genotype I of Class II ND viruses, has highly phylogenetic similarity to NDV strains isolated from waterfowl in China, but was distant from the viruses isolated in chickens and vaccine strains used in South Korea. Pathogenicity experiment in chickens revealed it to be a lentogenic virus. The deduced amino acid sequence of the cleavage site of the fusion (F) protein confirmed that the isolate contained the avirulent motif (112)GKQGRL(117) at the cleavage site and caused no apparent disease in chickens and ducks. With phylogeographic analysis based on fusion gene, we estimate the origin of an ancestral virus of the isolate and its sister strain located in China around 1998. It highlights the need of continuous surveillance to enhance current understanding of the molecular epidemiology and evolution of the pathogenic strains. PMID:26721461

  8. Characterization of a new Marburg virus isolated from a 1987 fatal case in Kenya.

    PubMed

    Johnson, E D; Johnson, B K; Silverstein, D; Tukei, P; Geisbert, T W; Sanchez, A N; Jahrling, P B

    1996-01-01

    In 1987, an isolated case of fatal Marburg disease was recognized during routine clinical haemorrhagic fever virus surveillance conducted in Kenya. This report describes the isolation and partial characterization of the new Marburg virus (strain Ravn) isolated from this case. The Ravn isolate was indistinguishable from reference Marburg virus strains by cross-neutralization testing. Virus particles and aggregates of Marburg nucleocapsid matrix in Ravn-infected vero cells, were visualized by immunoelectron microscopic techniques, and also in tissues obtained from the patient and from inoculated monkeys. The cell culture isolate produced a haemorrhagic disease typical of Marburg virus infection when inoculated into rhesus monkeys. Disease was characterized by the sudden appearance of fever and anorexia within 4 to 7 days, and death by day 11. Comparison of nucleotide sequences for portions of the glycoprotein genes of Marburg-Ravn were compared with Marburg reference strains Musoki (MUS) and Popp (POP). Nucleotide identity in this alignment between RAV and MUS is 72.3%, RAV and POP is 71%, and MUS and POP is 91.7%. Amino acid identity between RAV and MUS is 72%, RAV and POP is 67%, and MUS and POP is 93%. These data suggest that Ravn is another subtype of Marburg virus, analogous to the emerging picture of a spectrum of Ebola geographic isolates and subtypes. PMID:8800792

  9. Viruses isolated from Aedeomyia squamipennis mosquitoes collected in Panama, Ecuador, and Argentina: establishment of the Gamboa serogroup.

    PubMed

    Calisher, C H; Lazuick, J S; Justines, G; Francy, D B; Monath, T P; Gutierrez, E; Sabattini, M S; Bowen, G S; Jakob, W L

    1981-01-01

    Twenty-four virus strains were isolated from Aedeomyia squamipennis mosquitoes collected in Ecuador. One additional strain each was isolated from this species from Panama and ARgentina. All 26 isolates were shown to be related serologically to prototype Gamboa virus, originally isolated from Ad. squamipennis mosquitoes collected in Panama. Antigenic comparisons of eight strains, including prototype Gamboa virus, indicated the existence of four distinct viruses. Neutralization tests with sera from a variety of mammalian and avian species from Argentina provided further evidence that Gamboa serogroup viruses are transmitted between Ad. squamipennis and birds. PMID:6111232

  10. Isolation and genetic characterization of swinepox virus from pigs in India.

    PubMed

    Riyesh, Thachamvally; Barua, Sanjay; Kumar, Naveen; Jindal, Naresh; Bera, Bidhan Chandra; Narang, Gulshan; Mahajan, Nand Kishore; Arora, Devan; Anand, Taruna; Vaid, Rajesh Kumar; Yadav, Mansi; Chandel, Surender Singh; Malik, Praveen; Tripathi, Bhupendra Nath; Singh, Raj Kumar

    2016-06-01

    Swinepox virus (SWPV), a member of the genus Suipoxvirus causes generalized pock-like lesions on the body of domestic and wild pigs. Although outbreak has been reported in India since 1987, virus isolation and genetic characterization remained elusive. In September 2013, an outbreak of acute skin infection occurred in piglets in a commercial piggery unit at Rohtak district in Haryana, India. The presence of SWPV in scab samples collected from piglets succumbed to infection was confirmed by virus isolation, PCR amplification of SWPV-specific gene segments and nucleotide sequencing. Phylogenetic analysis of host-range genes of the SWPV revealed that the Indian isolate is genetically closely related to reference isolate SWPV/pig/U.S.A/1999/Nebraska. To the best of our knowledge this is the first report on isolation and genetic characterization of SWPV from pigs in India. PMID:27260812

  11. Isolation of cis-acting vaccinia virus DNA fragments promoting the expression of herpes simplex virus thymidine kinase by recombinant viruses.

    PubMed Central

    Vassef, A; Mars, M; Dru, A; Plucienniczak, A; Streeck, R E; Beaud, G

    1985-01-01

    Recombinant TK- vaccinia viruses containing the pBR322 sequence inserted in either orientation within the coding sequence of the viral thymidine kinase gene were constructed. They were characterized by genomic analysis, hybridization studies, reversion to wild-type virus by in vivo recombination, and rescue from their genomes of plasmids which contained all or parts of the pBR322 sequence. TK- cells were infected with one of these recombinant viruses and then transfected with pools of chimeric plasmids composed of a cloned herpes simplex virus thymidine kinase gene which contained upstream inserts of different vaccinia DNA fragments prepared by restriction or sonication. Recombination between homologous pBR322 sequences within infected cells generated selectable recombinant viruses in which expression of the herpes simplex virus thymidine kinase gene was promoted by the upstream vaccinia insert. These viruses were characterized by genomic analysis, hybridization, and in vivo or in vitro phosphorylation of (5-[125I]deoxycytidine as a specific assay for the expressed herpes simplex virus thymidine kinase. Vaccinia DNA inserts were isolated conveniently for transfer to bacteria by rescuing appropriate plasmids from the genome of recombinant viruses. The sequence of 100 nucleotides adjacent to the upstream region of the herpes simplex virus gene was determined in nine different inserts measuring 0.17 to 1.07 kilobase pairs. Images PMID:2989553

  12. Genetic and antigenic relatedness of H3 subtype influenza A viruses isolated from avian and mammalian species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Turkeys are susceptible to some swine influenza viruses based on natural and experimental transmissions of H1 and H3 subtype viruses from swine to turkeys. In 2004, we isolated triple reassortant H3N2 viruses from turkey breeder hens in Ohio and Illinois. These H3N2 viruses are currently the domin...

  13. Genetic and antigenic characterization of bovine viral diarrhea viruses isolated from cattle in Hokkaido, Japan

    PubMed Central

    ABE, Yuri; TAMURA, Tomokazu; TORII, Shiho; WAKAMORI, Shiho; NAGAI, Makoto; MITSUHASHI, Kazuya; MINE, Junki; FUJIMOTO, Yuri; NAGASHIMA, Naofumi; YOSHINO, Fumi; SUGITA, Yukihiko; NOMURA, Takushi; OKAMATSU, Masatoshi; KIDA, Hiroshi; SAKODA, Yoshihiro

    2015-01-01

    In our previous study, we genetically analyzed bovine viral diarrhea viruses (BVDVs) isolated from 2000 to 2006 in Japan and reported that subgenotype 1b viruses were predominant. In the present study, 766 BVDVs isolated from 2006 to 2014 in Hokkaido, Japan, were genetically analyzed to understand recent epidemics. Phylogenetic analysis based on nucleotide sequences of the 5′-untranslated region of viral genome revealed that 766 isolates were classified as genotype 1 (BVDV-1; 544 isolates) and genotype 2 (BVDV-2; 222). BVDV-1 isolates were further divided into BVDV-1a (93), 1b (371) and 1c (80) subgenotypes, and all BVDV-2 isolates were grouped into BVDV-2a subgenotype (222). Further comparative analysis was performed with BVDV-1a, 1b and 2a viruses isolated from 2001 to 2014. Phylogenetic analysis based on nucleotide sequences of the viral glycoprotein E2 gene, a major target of neutralizing antibodies, revealed that BVDV-1a, 1b and 2a isolates were further classified into several clusters. Cross-neutralization tests showed that BVDV-1b isolates were antigenically different from BVDV-1a isolates, and almost BVDV-1a, 1b and 2a isolates were antigenically similar among each subgenotype and each E2 cluster. Taken together, BVDV-1b viruses are still predominant, and BVDV-2a viruses have increased recently in Hokkaido, Japan. Field isolates of BVDV-1a, 1b and 2a show genetic diversity on the E2 gene with antigenic conservation among each subgenotype during the last 14 years. PMID:26400674

  14. Genetic and antigenic characterization of bovine viral diarrhea viruses isolated from cattle in Hokkaido, Japan.

    PubMed

    Abe, Yuri; Tamura, Tomokazu; Torii, Shiho; Wakamori, Shiho; Nagai, Makoto; Mitsuhashi, Kazuya; Mine, Junki; Fujimoto, Yuri; Nagashima, Naofumi; Yoshino, Fumi; Sugita, Yukihiko; Nomura, Takushi; Okamatsu, Masatoshi; Kida, Hiroshi; Sakoda, Yoshihiro

    2016-01-01

    In our previous study, we genetically analyzed bovine viral diarrhea viruses (BVDVs) isolated from 2000 to 2006 in Japan and reported that subgenotype 1b viruses were predominant. In the present study, 766 BVDVs isolated from 2006 to 2014 in Hokkaido, Japan, were genetically analyzed to understand recent epidemics. Phylogenetic analysis based on nucleotide sequences of the 5'-untranslated region of viral genome revealed that 766 isolates were classified as genotype 1 (BVDV-1; 544 isolates) and genotype 2 (BVDV-2; 222). BVDV-1 isolates were further divided into BVDV-1a (93), 1b (371) and 1c (80) subgenotypes, and all BVDV-2 isolates were grouped into BVDV-2a subgenotype (222). Further comparative analysis was performed with BVDV-1a, 1b and 2a viruses isolated from 2001 to 2014. Phylogenetic analysis based on nucleotide sequences of the viral glycoprotein E2 gene, a major target of neutralizing antibodies, revealed that BVDV-1a, 1b and 2a isolates were further classified into several clusters. Cross-neutralization tests showed that BVDV-1b isolates were antigenically different from BVDV-1a isolates, and almost BVDV-1a, 1b and 2a isolates were antigenically similar among each subgenotype and each E2 cluster. Taken together, BVDV-1b viruses are still predominant, and BVDV-2a viruses have increased recently in Hokkaido, Japan. Field isolates of BVDV-1a, 1b and 2a show genetic diversity on the E2 gene with antigenic conservation among each subgenotype during the last 14 years. PMID:26400674

  15. Characterization of A/eq-1 virus isolated during the equine influenza epidemic in India.

    PubMed

    Singh, G

    1994-02-01

    The equine influenza virus, Ludhiana/5/87, isolated from the clinical material during the epidemic of equine influenza in India in 1987 was inhibited in haemagglutination-inhibition test by the antiserum against the prototype A/eq/Prague/1/56 (H7N7) virus and by post-epidemic horse sera. In haemagglutinin and neuraminidase analysis, the A/eq/Ludhiana/5/87 isolate appeared similar to the prototype A/eq/Prague/1/56 virus and was characterized as the H7N7 subtype. PMID:8067310

  16. Genomic and phylogenetic characterization of Merino Walk virus, a novel arenavirus isolated in South Africa

    PubMed Central

    Palacios, Gustavo; Savji, Nazir; Hui, Jeffrey; Travassos da Rosa, Amelia; Popov, Vsevolod; Briese, Thomas; Tesh, Robert; Lipkin, W. Ian

    2010-01-01

    Merino Walk virus (MWV), a proposed novel tentative species of the family Arenaviridae, was isolated from a rodent, Myotomys unisulcatus, collected at Merino Walk, Eastern Cape, South Africa, in 1985. Full-length genomic sequence confirmed MWV as an arenavirus related distantly to Mobala, Mopeia and Ippy viruses, all members of the Old World arenavirus complex. We propose MWV as a tentative novel species in the Lassa–lymphocytic choriomeningitis virus complex, based on its isolation from a novel rodent species and its genetic and serological characteristics. PMID:20071489

  17. Characterisation of an isolate of Narcissus degeneration virus from Chinese narcissus (Narcissus tazetta var. chinensis).

    PubMed

    Chen, J; Shi, Y-H; Adams, M J; Zheng, H-Y; Qin, B-X; Chen, J-P

    2007-02-01

    A potyvirus from Chinese narcissus was transmitted mechanically to three species of Narcissus and to Lycoris radiata but not to 22 other test species. In western blot, the coat protein reacted strongly with Narcissus degeneration virus (UK isolate) antiserum. Antiserum raised to the Chinese virus did not react with eighteen other potyviruses. The complete nucleotide sequence (9816 nt) had the typical genome organisation for a member of the genus Potyvirus. Sequence comparisons and phylogenetic analysis showed that the Chinese virus was different from all previously sequenced potyviruses but distantly related to onion yellow dwarf and shallot yellow stripe viruses. PMID:16932980

  18. The complete sequence of the genomic RNA of an isolate of Lily virus X (genus Potexvirus).

    PubMed

    Chen, J; Shi, Y-H; Adams, M J; Chen, J-P

    2005-04-01

    The complete sequence of the genomic RNA of an isolate of Lily virus X (LVX) has been determined for the first time. The isolate from the Netherlands was 5823 nucleotide (nt) long excluding the 3'-poly(A) tail, making it the shortest reported potexvirus sequence. The 5'-non-coding region begins with GGAAAA like that of Scallion virus X (ScaVX) and some isolates of Cymbidium mosaic virus (CymMV), whereas those of other sequenced potexviruses probably all begin with GAAAA. The genome organisation was similar to that of other members of the genus except that a TGBp3-like region lacked a normal AUG start codon. A phylogenetic analysis based on the entire coding sequence showed that LVX was most closely related to Strawberry mild yellow edge virus and belonged in a subgroup of the genus that also contains CymMV, Narcissus mosaic virus, ScaVX, Pepino mosaic virus, Potato aucuba mosaic virus and White clover mosaic virus. PMID:15578239

  19. Archaeal Viruses of the Sulfolobales: Isolation, Infection, and CRISPR Spacer Acquisition.

    PubMed

    Erdmann, Susanne; Garrett, Roger A

    2015-01-01

    Infection of archaea with phylogenetically diverse single viruses, performed in different laboratories, has failed to activate spacer acquisition into host CRISPR loci. The first successful uptake of archaeal de novo spacers was observed on infection of Sulfolobus solfataricus P2 with an environmental virus mixture isolated from Yellowstone National Park (Erdmann and Garrett, Mol Microbiol 85:1044-1056, 2012). Experimental studies of isolated genetic elements from this mixture revealed that SMV1 (S ulfolobus Monocauda Virus 1), a tailed spindle-shaped virus, can induce spacer acquisition in CRISPR loci of Sulfolobus species from a second coinfecting conjugative plasmid or virus (Erdmann and Garrett, Mol Microbiol 85:1044-1056, 2012; Erdmann et al. Mol Microbiol 91:900-917, 2014). Here we describe, firstly, the isolation of archaeal virus mixtures from terrestrial hot springs and the techniques used both to infect laboratory strains with these virus mixtures and to obtain purified virus particles. Secondly, we present the experimental conditions required for activating SMV1-induced spacer acquisition in two different Sulfolobus species. PMID:25981476

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

    PubMed

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

    2014-10-01

    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

  1. Phylogenetic and antigenic characterization of newly isolated porcine epidemic diarrhea viruses in Japan.

    PubMed

    Islam, Md Taimur; Kubota, Tomoe; Ujike, Makoto; Yahara, Yoshiriro; Taguchi, Fumihiro

    2016-08-15

    To evaluate the mechanism by which a large outbreak of porcine epidemic diarrhea (PED) occurred in Japan, where the majority of sows are vaccinated, we isolated two new strains of PED virus (PEDV) from the intestines of piglets and found that they showed greater similarity to US isolates (group II PEDV) than to the Japanese vaccine strain (group I PEDV). We compared the antigenicity of the vaccine type strain and newly isolated strains by means of a neutralization test using sera from a number of pigs from various farms; the results revealed that they are antigenically similar. This is the first report of the similarity of group I and II viruses using sera from individual pigs vaccinated with group I virus. These data suggest that the large outbreak of PED in Japan cannot be attributed to inefficient vaccination but may be due to the extremely high virulence of the newly appearing viruses. PMID:27292080

  2. Nhumirim virus, a novel flavivirus isolated from mosquitoes from the Pantanal, Brazil

    PubMed Central

    Solberg, Owen; Couto-Lima, Dinair; Kenney, Joan; Serra-Freire, Nicolau; Brault, Aaron; Nogueira, Rita; Langevin, Stanley; Komar, Nicholas

    2016-01-01

    We describe the isolation of a novel flavivirus, isolated from a pool of mosquitoes identified as Culex (Culex) chidesteri collected in 2010 in the Pantanal region of west-central Brazil. The virus is herein designated Nhumirim virus (NHUV) after the name of the ranch from which the mosquito pool was collected. Flavivirus RNA was detected by real-time RT-PCR of homogenized mosquitoes and from the corresponding C6/36 culture supernatant. Based on full-genome sequencing, the virus isolate was genetically distinct from but most closely related to Barkedji virus (BJV), a newly described flavivirus from Senegal. Phylogenetic analysis demonstrated that NHUV grouped with mosquito-borne flaviviruses forming a clade with BJV. This clade may be genetically intermediate between the Culex-borne flaviviruses amplified by birds and the insect-only flaviviruses. PMID:25252815

  3. Genome Sequences of Nine Vesicular Stomatitis Virus Isolates from South America

    PubMed Central

    King, David J.; Howson, Emma L. A.; Madi, Mikidache; Pauszek, Steven J.; Rodriguez, Luis L.; Knowles, Nick J.; Mioulet, Valérie; King, Donald P.

    2016-01-01

    We report nine full-genome sequences of vesicular stomatitis virus obtained by Illumina next-generation sequencing of RNA, isolated from either cattle epithelial suspensions or cell culture supernatants. Seven of these viral genomes belonged to the New Jersey serotype/species (clade III), while two isolates belonged to the Indiana serotype/species. PMID:27081129

  4. Complete Genome Sequence of Alternanthera mosaic virus, Isolated from Achyranthes bidentata in Asia

    PubMed Central

    Iwabuchi, Nozomu; Yoshida, Tetsuya; Yusa, Akira; Nishida, Shuko; Tanno, Kazuyuki; Keima, Takuya; Nijo, Takamichi; Yamaji, Yasuyuki

    2016-01-01

    Alternanthera mosaic virus (AltMV) infecting Achyranthes bidentata was first detected in Asia, and the complete genome sequence (6,604 nucleotides) was determined. Sequence identity analysis and phylogenetic analysis confirmed that this isolate is the most phylogenetically distant AltMV isolate worldwide. PMID:26988034

  5. Genetic Detection and Isolation of Crimean-Congo hemorrhagic fever virus, Kosovo, Yugoslavia

    PubMed Central

    Boźović, Bojana; Pavlidou, Vassiliki; Papadimitriou, Evangelia; Pelemis, Mijomir; Antoniadis, Aantonis

    2002-01-01

    Crimean-Congo hemorrhagic fever virus (C-CHFV) strains were isolated from a fatal case and the attending physician in Kosovo, Yugoslavia. Early, rapid diagnosis of the disease was achieved by reverse transcription-polymerase chain reaction. The physician was successfully treated with oral ribavirin. These cases yielded the first genetically studied C-CHFV human isolates in the Balkans. PMID:12141973

  6. Biological and molecular characterization of a reticuloendotheliosis virus isolated from turkeys with lymphomas

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two reticuloendotheliosis virus (REV) isolates termed AF-140-11 and AF-140-12 were obtained from turkeys with increased mortality, disseminated lymphoblastoid neoplasia, and decreased egg production. The REV isolates were propagated and titrated in chicken-embryo fibroblasts (CEFs) obtained from a s...

  7. Phylogenetic Characterization of Genotype 4 Hepatitis C Virus Isolates from Argentina

    PubMed Central

    Alfonso, Victoria; Flichman, Diego; Sookoian, Silvia; Mbayed, Viviana Andrea; Campos, Rodolfo Héctor

    2001-01-01

    Among 114 patients infected with hepatitis C virus, three genotype 4 isolates, unusual in Argentina, were detected by phylogenetic analysis over different genomic regions. The patients were not related. One sample was associated with Egyptian sequences, and the others were associated with a Zairean isolate, a fact which reinforces the idea that they are from independent sources. PMID:11326031

  8. Complete nucleotide sequence of a maize chlorotic mottle virus isolate from Nebraska

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The complete genome of a maize chlorotic mottle virus isolate from Nebraska (MCMV-NE) was cloned and sequenced. The MCMV-NE genome consists of 4,436 nucleotides and shares 99.5% nucleotide sequence identity with an MCMV isolate from Kansas (MCMV-KS). Of 22 polymorphic sites, most resulted from t...

  9. Genome Sequences of Nine Vesicular Stomatitis Virus Isolates from South America.

    PubMed

    Fowler, Veronica L; King, David J; Howson, Emma L A; Madi, Mikidache; Pauszek, Steven J; Rodriguez, Luis L; Knowles, Nick J; Mioulet, Valérie; King, Donald P

    2016-01-01

    We report nine full-genome sequences of vesicular stomatitis virus obtained by Illumina next-generation sequencing of RNA, isolated from either cattle epithelial suspensions or cell culture supernatants. Seven of these viral genomes belonged to the New Jersey serotype/species (clade III), while two isolates belonged to the Indiana serotype/species. PMID:27081129

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The complete genome sequence of an isolate of Grapevine virus E (GVE) collected from a red-berried wine grape cultivar (Cabernet Sauvignon) in Washington State was determined. The 7,568 nt long genome of GVE is similar in size and sequence identity with a GVE isolate from a wine grape cv. Shiraz fro...

  11. Population Structure of Blueberry Mosaic Associated Virus: Evidence of Genetic Exchange in Geographically Distinct Isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The population structure of blueberry mosaic associated virus (BlMaV), a putative member of the family Ophioviridae, was examined using 59 isolates collected from North America and Slovenia. The studied isolates displayed low genetic diversity in the movement and nucleoprotein regions and low ratios...

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

    USGS Publications Warehouse

    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

    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.

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

    PubMed Central

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

    2016-01-01

    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

  14. Massilia Virus, A Novel Phlebovirus (Bunyaviridae) Isolated from Sandflies in the Mediterranean

    PubMed Central

    Moureau, Grégory; Temmam, Sarah; Izri, Arezki; Marty, Pierre; Parola, Philippe; da Rosa, Amelia Travassos; Tesh, Robert B.; de Lamballerie, Xavier

    2009-01-01

    Abstract A new virus was isolated from three independent pools of Phlebotomus perniciosus sandflies (Diptera; Psychodidae) trapped in two regions of southeastern France, located 90 miles apart. Microscopic, antigenic and genetic analyses indicate that this novel virus belongs to the genus Phlebovirus in the family Bunyaviridae. The new virus is designated Massilia virus since the first isolate was obtained from sandflies collected in the suburban area of Marseille. The complete genome sequence was determined and used to compare the genetic and phylogenetic relationships of Massilia virus with other phleboviruses. Genetic and antigenic properties were employed to address whether or not Massilia virus should be considered a new species within the genus, or a member of a previously recognized species. Cerebrospinal fluid specimens, collected from local patients with central nervous system infections during the previous four-year period were tested for the presence of Massilia virus RNA, but gave negative results. In conclusion, Massilia virus is proposed as a member of the Sand-fly fever Naples virus complex; its public health importance has yet to be determined. PMID:19055373

  15. How Does Sampling Methodology Influence Molecular Detection and Isolation Success in Influenza A Virus Field Studies?

    PubMed

    Latorre-Margalef, Neus; Avril, Alexis; Tolf, Conny; Olsen, Björn; Waldenström, Jonas

    2016-02-01

    Wild waterfowl are important reservoir hosts for influenza A virus (IAV) and a potential source of spillover infections in other hosts, including poultry and swine. The emergence of highly pathogenic avian influenza (HPAI) viruses, such as H5N1 and H5N8, and subsequent spread along migratory flyways prompted the initiation of several programs in Europe, North America, and Africa to monitor circulation of HPAI and low-pathogenicity precursor viruses (low-pathogenicity avian influenza [LPAI] viruses). Given the costs of maintaining such programs, it is essential to establish best practice for field methodologies to provide robust data for epidemiological interpretation. Here, we use long-term surveillance data from a single site to evaluate the influence of a number of parameters on virus detection and isolation of LPAI viruses. A total of 26,586 samples (oropharyngeal, fecal, and cloacal) collected from wild mallards were screened by real-time PCR, and positive samples were subjected to isolation in embryonated chicken eggs. The LPAI virus detection rate was influenced by the sample type: cloacal/fecal samples showed a consistently higher detection rate and lower cycle threshold (Ct) value than oropharyngeal samples. Molecular detection was more sensitive than isolation, and virus isolation success was proportional to the number of RNA copies in the sample. Interestingly, for a given Ct value, the isolation success was lower in samples from adult birds than in those from juveniles. Comparing the results of specific real-time reverse transcriptase (RRT)-PCRs and of isolation, it was clear that coinfections were common in the investigated birds. The effects of sample type and detection methods warrant some caution in interpretation of the surveillance data. PMID:26655759

  16. How Does Sampling Methodology Influence Molecular Detection and Isolation Success in Influenza A Virus Field Studies?

    PubMed Central

    Avril, Alexis; Tolf, Conny; Olsen, Björn

    2015-01-01

    Wild waterfowl are important reservoir hosts for influenza A virus (IAV) and a potential source of spillover infections in other hosts, including poultry and swine. The emergence of highly pathogenic avian influenza (HPAI) viruses, such as H5N1 and H5N8, and subsequent spread along migratory flyways prompted the initiation of several programs in Europe, North America, and Africa to monitor circulation of HPAI and low-pathogenicity precursor viruses (low-pathogenicity avian influenza [LPAI] viruses). Given the costs of maintaining such programs, it is essential to establish best practice for field methodologies to provide robust data for epidemiological interpretation. Here, we use long-term surveillance data from a single site to evaluate the influence of a number of parameters on virus detection and isolation of LPAI viruses. A total of 26,586 samples (oropharyngeal, fecal, and cloacal) collected from wild mallards were screened by real-time PCR, and positive samples were subjected to isolation in embryonated chicken eggs. The LPAI virus detection rate was influenced by the sample type: cloacal/fecal samples showed a consistently higher detection rate and lower cycle threshold (Ct) value than oropharyngeal samples. Molecular detection was more sensitive than isolation, and virus isolation success was proportional to the number of RNA copies in the sample. Interestingly, for a given Ct value, the isolation success was lower in samples from adult birds than in those from juveniles. Comparing the results of specific real-time reverse transcriptase (RRT)-PCRs and of isolation, it was clear that coinfections were common in the investigated birds. The effects of sample type and detection methods warrant some caution in interpretation of the surveillance data. PMID:26655759

  17. Neutralizing monoclonal antibodies recognize antigenic variants among isolates of infectious hematopoietic necrosis virus

    USGS Publications Warehouse

    Winton, J.R.; Arakawa, C.N.; Lannan, C.N.; Fryer, J.L.

    1988-01-01

    eutralizing monoclonal antibodies were developed against strains of infectious hematopoietic necrosis virus (IHNV) from steelhead trout Salmo gairdneri in the Deschutes River of Oregon, chinook salmon Oncorhynchus tshawytscha in the Sacramento River of California, and rainbow trout Salmo gairdneri reared in the Hagerman Valley of Idaho, USA. These antibodies were tested for neutralization of 12 IHNV isolates obtained from salmonids in Japan, Alaska, Washington, Oregon, California, and Idaho. The antibodies recognized antigenic variants among the isolates and could be used to separate the viruses into 4 groups. The members of each group tended to be related by geographic area rather than by source host species, virulence, or date of isolation.

  18. Analysis of infectious laryngotracheitis virus isolates from Ontario and New Brunswick by the polymerase chain reaction.

    PubMed Central

    Alexander, H S; Key, D W; Nagy, E

    1998-01-01

    The polymerase chain reaction (PCR) was used to amplify DNA of infectious laryngotracheitis virus (ILTV) isolates obtained from field specimens. The examined 47 samples included 37 isolates representing 35 cases of infectious laryngotracheitis from Ontario and 10 isolates originating from 10 field cases in New Brunswick. The viruses were grown in either embryonated chicken eggs or cell culture, the DNA extracted and amplified using primers designed from the sequence information of a 1.1 kb BamHI fragment of the Ontario 1598 ILTV strain. Thirty-four of the Ontario isolates and all of the New Brunswick isolates were amplified successfully. This suggests that the selected primers would be useful for the majority of the isolates encountered in outbreaks of ILTV. Images Figure 1. Figure 2. PMID:9442943

  19. Rapid isolation of herpes simplex virus by using mink lung and rhabdomyosarcoma cell cultures.

    PubMed Central

    Johnston, S L; Wellens, K; Siegel, C S

    1990-01-01

    Highly sensitive and rapid results can be obtained by isolating herpes simplex virus from clinical specimens in simple cell culture with rhabdomyosarcoma (RD) cells. In this study, 3,186 clinical specimens were inoculated into locally produced, equivalent-age RD and mink lung (ML) cells. Of 727 positive isolates, all (100%) were isolated from RD cells and only 691 (95%) were isolated from ML cells. Furthermore, 162 of the positive isolates (22%) were isolated in RD cells earlier than in ML cells. RD cells are continuous and can be cultivated in house without decreasing sensitivity as the passage number increases. They produce a highly distinguishable cytopathic effect in response to herpes simplex virus and maintain intense confirmatory staining patterns. PMID:2177754

  20. Characterization of West Nile viruses isolated form captive American flamingoes (Phoenicopterus ruber) in Medellin, Colombia.

    USGS Publications Warehouse

    Osorio, Jorge E.; Ciuoderis, Karl A.; Lopera, Juan G.; Piedrahita, Leidy D.; Murphy, Darby; LeVasseur, James; Carrillo, Lina; Ocampo, Martha C.; Hofmeister, Erik

    2012-01-01

    Serum samples from a total of 71 healthy captive birds belonging to 18 species were collected in July of 2008 in Medellin (Colombia) and tested for flaviviruses. Eighteen of 29 samples from American Flamingoes (Phoenicopterus ruber) were positive for West Nile virus (WNV) by reverse transcription-polymerase chain reaction. Selected positive samples were serially passaged and WNV was confirmed by immunofluorescence. Two isolates (524/08, 9835/08) were characterized in vitro and in vivo. Sequence analysis revealed WNV with 16 nucleotide substitutions resulting in six amino acid changes when compared with the NY99 strain. Colombian (COL) viruses were more closely related to Louisiana isolates from 2001. When compared with attenuated strains isolated from Texas, COL isolates differed in their plaque size and temperature sensitivity phenotype. The COL viruses were pathogenic in embryonated chicken eggs and Balb/c mice.

  1. Characterization of West Nile Viruses Isolated from Captive American Flamingoes (Phoenicopterus ruber) in Medellin, Colombia

    PubMed Central

    Osorio, Jorge E.; Ciuoderis, Karl A.; Lopera, Juan G.; Piedrahita, Leidy D.; Murphy, Darby; LeVasseur, James; Carrillo, Lina; Ocampo, Martha C.; Hofmeister, Erik

    2012-01-01

    Serum samples from a total of 71 healthy captive birds belonging to 18 species were collected in July of 2008 in Medellin (Colombia) and tested for flaviviruses. Eighteen of 29 samples from American Flamingoes (Phoenicopterus ruber) were positive for West Nile virus (WNV) by reverse transcription-polymerase chain reaction. Selected positive samples were serially passaged and WNV was confirmed by immunofluorescence. Two isolates (524/08, 9835/08) were characterized in vitro and in vivo. Sequence analysis revealed WNV with 16 nucleotide substitutions resulting in six amino acid changes when compared with the NY99 strain. Colombian (COL) viruses were more closely related to Louisiana isolates from 2001. When compared with attenuated strains isolated from Texas, COL isolates differed in their plaque size and temperature sensitivity phenotype. The COL viruses were pathogenic in embryonated chicken eggs and Balb/c mice. PMID:22802436

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

    PubMed Central

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

    2012-01-01

    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

  3. Herpes simplex virus latency in isolated human neurons.

    PubMed Central

    Wigdahl, B; Smith, C A; Traglia, H M; Rapp, F

    1984-01-01

    Herpes simplex virus is most probably maintained in the ganglion neurons of the peripheral nervous system of humans in a latent form that can reactivate to produce recurrent disease. As an approximation of this cell-virus interaction, we have constructed a herpes simplex virus latency in vitro model system using human fetus sensory neurons as the host cell. Human fetus neurons were characterized as neuronal in origin by the detection of the neuropeptide substance P and the neuron-specific plasma membrane A2B5 antigen. Virus latency was established by blocking complete expression of the virus genome by treatment of infected human neurons with a combination of human leukocyte interferon and (E)-5-(2-bromovinyl)-2'-deoxyuridine for 7 days. After removal of inhibitors, virus latency was maintained for at least 9 days. This in vitro model will provide a system to analyze, in a primary human neuron, the state of the herpes simplex virus genome during establishment and maintenance of experimental latency. Images PMID:6091142

  4. Complete Genome Sequences of Three Clinical Isolates of Dengue Virus Serotype 1 from South Korean Travelers

    PubMed Central

    Jung, Eunhye; Jeong, Young Eui; Balasuriya, Udeni B. R.

    2015-01-01

    In this study, we report the complete genome sequences of three clinical isolates of dengue virus serotype 1 isolated from South Korean travelers returning from different countries in Southeast Asia. The nucleotide sequence identities ranged from 91.5 to 92.2%, while the amino acid sequence identities ranged from 97.5 to 97.9% among the three clinical isolates. PMID:26607895

  5. Genetic characterization of dengue virus type 1 isolated in Brunei in 2005-2006.

    PubMed

    Osman, Osmali; Fong, Mun Yik; Sekaran, Shamala Devi

    2009-03-01

    The full-length genomes of two DENV-1 viruses isolated during the 2005-2006 dengue incidents in Brunei were sequenced. Twenty five primer sets were designed to amplify contiguous overlapping fragments of approximately 500-600 base pairs spanning the entire sequence of the genome. The amplified PCR products were sent to a commercial laboratory for sequencing and the nucleotides and the deduced amino acids were determined. Sequence analysis of the envelope gene at the nucleotide and amino acid levels between the two isolates showed 92 and 96 % identity, respectively. Comparison of the envelope gene sequences with 68 other DENV-1 viruses of known genotypes placed the two isolates into two different genotypic groups. Isolate DS06/210505 belongs to genotype V together with some of the recent isolates from India (2003) and older isolates from Singapore (1990) and Burma (1976), while isolate DS212/110306 was clustered in genotype IV with the prototype Nauru strain (1974) and with some of the recent isolates from Indonesia (2004) and the Philippines (2002, 2001). In the full-length genome analysis at the nucleotide level, isolate DS06/210505 showed 94 % identity to the French Guyana strain (1989) in genotype V while isolate DS212/110306 had 96 % identity to the Nauru Island strain (1974) in genotype IV. This work constitutes the first complete genetic characterization of not only Brunei DENV-1 virus isolates, but also the first strain from Borneo Island. This study was the first to report the isolation of dengue virus in the country. PMID:19218214

  6. Amantadine resistance among highly pathogenic avian influenza viruses (H5N1) isolated from India.

    PubMed

    Jacob, Aron; Sood, Richa; Chanu, Kh Victoria; Bhatia, Sandeep; Khandia, Rekha; Pateriya, A K; Nagarajan, S; Dimri, U; Kulkarni, D D

    2016-02-01

    Emergence of antiviral resistance among H5N1 avian influenza viruses is the major challenge in the control of pandemic influenza. Matrix 2 (M2) inhibitors (amantadine and rimantadine) and neuraminidase inhibitors (oseltamivir and zanamivir) are the two classes of antiviral agents that are specifically active against influenza viruses and are used for both treatment and prophylaxis of influenza infections. Amantadine targets the M2 ion channel of influenza A virus and interrupts virus life cycle through blockade of hydrogen ion influx. This prevents uncoating of the virus in infected host cells which impedes the release of ribonucleoprotein required for transcription and replication of virion in the nucleus. The present study was carried out to review the status of amantadine resistance in H5N1 viruses isolated from India and to study their replicative capability. Results of the study revealed resistance to amantadine in antiviral assay among four H5N1 viruses out of which two viruses had Serine 31 Asparagine (AGT-AAT i.e., S31N) mutation and two had Valine 27 Alanine (GTT-GCT i.e., V27A) mutation. The four resistant viruses not only exhibited significant difference in effective concentration 50% (EC50) values of amantadine hydrochloride from that of susceptible viruses (P < 0.0001) but also showed significant difference between two different types (S31N and V27A) of mutant viruses (P < 0.05). Resistance to amantadine could also be demonstrated in a simple HA test after replication of the viruses in MDCK cells in presence of amantadine. The study identifies the correlation between in vitro antiviral assay and presence of established molecular markers of resistance, the retention of replicative capacity in the presence of amantadine hydrochloride by the resistant viruses and the emergence of resistant mutations against amantadine among avian influenza viruses (H5N1) without selective drug pressure. PMID:26639679

  7. A genetically novel, narrow-host-range isolate of cucumber mosaic virus (CMV) from rosemary.

    PubMed

    Tepfer, Mark; Girardot, Gregory; Fénéant, Lucie; Ben Tamarzizt, Hana; Verdin, Eric; Moury, Benoît; Jacquemond, Mireille

    2016-07-01

    An isolate of cucumber mosaic virus (CMV), designated CMV-Rom, was isolated from rosemary (Rosmarinus officinalis) plants in several locations near Avignon, France. Laboratory studies showed that, unlike typical CMV isolates, CMV-Rom has a particularly narrow host range. It could be transmitted by aphids Aphis gossypii and Myzus persicae, but with low efficacy compared to a typical CMV isolate. Phylogenetic analysis of the nucleotide sequences of the CMV-Rom genomic RNAs shows that this isolate does not belong to any of the previously described CMV subgroups, IA, IB, II or III. PMID:27138549

  8. Molecular detection and analysis of Sheeppox and Orf viruses isolated from sheep from Qalubia, Egypt.

    PubMed

    Selim, Abdelfattah; Elhaig, Mahmoud; Höche, Jennifer; Gaede, Wolfgang

    2016-01-01

    In this study an outbreak with Sheeppox virus (SPPV) and Orf virus (ORFV) in one sheep herd in the Qalubia province, Egypt, was investigated. Both, SPPV and ORFV caused clinically manifest infections among sheep. The affected sheep showed skin lesions around the mouth or all over the body. Therefore, reliable diagnosis should confirm the aetiology of the infection and then reduce spread of the diseases in the affected areas. Clinical samples were investigated by virus isolation, PCR and real-time PCR assays. Furthermore, PCR-products of SPPV and ORFV isolates were sequenced and alignment to reference isolates was performed for phylogenetic analyses. The laboratory diagnosis showed that real-time PCR assay was more accurate and sensitive than conventional PCR and virus isolation. In phylogenetic analysis of the A29L gene genetic differences between SPPV field strains were not observed and the strains showed 100% homology with two SPPV isolates from Kazakhstan and one isolate from Turkey. The ORFV field strains are in the P55 gene genetically distinct from another and from other published isolates from Egypt 2006 and 2009. PMID:27529993

  9. A molecular epidemiological study targeting the glycoprotein gene of rabies virus isolates from China.

    PubMed

    Meng, Sheng-Li; Yan, Jia-Xin; Xu, Ge-Lin; Nadin-Davis, Susan A; Ming, Ping-Gang; Liu, Sheng-Ya; Wu, Jie; Ming, He-Tian; Zhu, Feng-Cai; Zhou, Dun-Jin; Xiao, Qi-You; Dong, Guan-Mu; Yang, Xiao-Ming

    2007-03-01

    A group of 31 rabies viruses (RABVs), recovered primarily from dogs, one deer and one human case, were collected from various areas in China between 1989 and 2006. Complete G gene sequences determined for these isolates indicated identities of nucleotide and amino acid sequences of >or=87% and 93.8%, respectively. Phylogenetic analysis of these and some additional Chinese isolates clearly supported the placement of all Chinese viruses in Lyssavirus genotype 1 and divided all Chinese isolates between four distinct groups (I-IV). Several variants identified within the most commonly encountered group I were distributed according to their geographical origins. A comparison of representative Chinese viruses with other isolates retrieved world-wide indicated a close evolutionary relationship between China group I and II viruses and those of Indonesia while China group III viruses formed an outlying branch to variants from Malaysia and Thailand. China group IV viruses were closely related to several vaccine strains. The predicted glycoprotein sequences of these RABVs variants are presented and discussed with respect to the utility of the anti-rabies biologicals currently employed in China. PMID:17129631

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

    PubMed Central

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

    1993-01-01

    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

  11. [Complete genomic analysis of a novel infectious bronchitis virus isolate].

    PubMed

    Hu, Bei-Xia; Yang, Shao-Hua; Zhang, Xiu-Mei; Zhang, Wei; Cao, San-Jie; Xu, Chuan-Tian; Huang, Qing-Hua; Zhang, Lin; Huang, Yan-Yan; Wen, Xin-Tian

    2014-07-01

    The genome of CK/CH/SD09/005, an isolate of infectious bronchitis virus (IBV), was characterized to enable the further understanding of the epidemiology and evolution of IBV in China. Twenty-five pairs of primers were designed to amplify the full-length genome of CK/CH/SD09/005. The nucleotide sequence of CK/CH/SD09/005 was compared with reference IBV strains retrieved from GenBank. The phylogenic relationship between CK/CH/SD09/005 and the reference strains was analyzed based on S1 gene sequences. The complete genome of CK/CH/SD09/005 consisted of 27691 nucleotides (nt), excluding the 5' cap and 3' poly A tail. The whole-genome of CK/CH/SD09/005 shared 97 - 99% nucleotide sequence homology with the GX-NN09032 strain, which was the only complete genome that was closely related to CK/CH/SD09/005. When compared with all reference strains except GX-NN09032, CK/CH/SD09/005 showed the highest similarity to ck/CH/LDL/091022 and SDIB821/2012 (QX-like) in the replicase gene (Gene 1) and 3'UTR, with a sequence identity rate of 97% and 98%, respectively. However, CK/CH/SD09/005 exhibited lower levels of similarity with ck/CH/LDL/091022 and SDIB821/2012 in S-3a-3b-3c/ E-M-5a-5b-N with a sequence identity of 72% - 90%. CK/CH/SD09/005 showed the highest level of nucleotide identity with Korean strain 1011, and Chinese strains CK/CH/LXJ/02I, DK/CH/HN/ZZ2004 and YX10, in ORF 3c/E (97%), 5a (96%), 5b (99%) and N (96%), respectively. ORFs 3a, 3b and M of CK/CH/SD09/005 exhibited no more than 90% homology with the reference strains, excluding GX-NN09032. The phylogenic analysis based on the S1 gene revealed that CK/CH/SD09/005 and 39 published strains were classified into seven clades (genotypes). CK/CH/SD09/005 was distributed in clade IV with several isolates collected between 2007 and 2012. CK/CH/SD09/005 showed 66% - 69% and 72% - 81% nucleotide identities with the IBV strains of other six clades in the S1 and S2 subunits, respectively. More over, multiple substitutions were

  12. Isolation of novel triple‐reassortant swine H3N2 influenza viruses possessing the hemagglutinin and neuraminidase genes of a seasonal influenza virus in Vietnam in 2010

    PubMed Central

    Ngo, Long Thanh; Hiromoto, Yasuaki; Pham, Vu Phong; Le, Ha Thi Hong; Nguyen, Ha Truc; Le, Vu Tri; Takemae, Nobuhiro; Saito, Takehiko

    2011-01-01

    Please cite this paper as: Ngo et al. (2012) Isolation of novel triple‐reassortant swine H3N2 influenza viruses possessing the hemagglutinin and neuraminidase genes of a seasonal influenza virus in Vietnam in 2010. Influenza and Other Respiratory Viruses 6(1), 6–10. Surveillance of swine influenza viruses (SIVs) in 31 pig farms in northern and southern parts of Vietnam was conducted. Six H3N2 influenza A viruses were isolated from a pig farm in southern Vietnam. They were novel genetic reassortants between a triple–reassortant SIV and a human seasonal H3N2 virus. Their hemagglutinin and neuraminidase genes were derived from a human virus circulating around 2004–2006 and the remaining genes from a triple‐reassortant SIV that originated in North America. This is the first report describing the isolation of a novel triple‐reassortant SIV in Vietnam. PMID:21668659

  13. Characterization of influenza A (H7N9) viruses isolated from human cases imported into Taiwan.

    PubMed

    Yang, Ji-Rong; Kuo, Chuan-Yi; Huang, Hsiang-Yi; Wu, Fu-Ting; Huang, Yi-Lung; Cheng, Chieh-Yu; Su, Yu-Ting; Wu, Ho-Sheng; Liu, Ming-Tsan

    2015-01-01

    A novel avian influenza A (H7N9) virus causes severe human infections and was first identified in March 2013 in China. The H7N9 virus has exhibited two epidemiological peaks of infection, occurring in week 15 of 2013 and week 5 of 2014. Taiwan, which is geographically adjacent to China, faces a large risk of being affected by this virus. Through extensive surveillance, launched in April 2013, four laboratory-confirmed H7N9 cases imported from China have been identified in Taiwan. The H7N9 virus isolated from imported case 1 in May 2013 (during the first wave) was found to be closest genetically to a virus from wild birds and differed from the prototype virus, A/Anhui/1/2013, in the MP gene. The other three imported cases were detected in December 2013 and April 2014 (during the second wave). The viruses isolated from cases 2 and 4 were similar in the compositions of their 6 internal genes and distinct from A/Anhui/1/2013 in the PB2 and MP genes, whereas the virus isolated from case 3 exhibited a novel reassortment that has not been identified previously and was different from A/Anhui/1/2013 in the PB2, PA and MP genes. The four imported H7N9 viruses share similar antigenicity with A/Anhui/1/2013, and their HA and NA genes grouped together in their respective phylogenies. In contrast with the HA and NA genes, which exhibited a smaller degree of diversity, the internal genes were heterogeneous and provided potential distinctions between transmission sources in terms of both geography and hosts. It is important to strengthen surveillance of influenza and to share viral genetic data in real-time for reducing the threat of rapid and continuing evolution of H7N9 viruses. PMID:25748033

  14. Antiviral susceptibility of influenza viruses isolated from patients pre- and post-administration of favipiravir.

    PubMed

    Takashita, Emi; Ejima, Miho; Ogawa, Rie; Fujisaki, Seiichiro; Neumann, Gabriele; Furuta, Yousuke; Kawaoka, Yoshihiro; Tashiro, Masato; Odagiri, Takato

    2016-08-01

    Favipiravir, a viral RNA-dependent RNA polymerase inhibitor, has recently been approved in Japan for influenza pandemic preparedness. Here, we conducted a cell-based screening system to evaluate the susceptibility of influenza viruses to favipiravir. In this assay, the antiviral activity of favipiravir is determined by inhibition of virus-induced cytopathic effect, which can be measured by using a colorimetric cell proliferation assay. To demonstrate the robustness of the assay, we compared the favipiravir susceptibilities of neuraminidase (NA) inhibitor-resistant influenza A(H1N1)pdm09, A(H3N2), A(H7N9) and B viruses and their sensitive counterparts. No significant differences in the favipiravir susceptibilities were found between NA inhibitor-resistant and sensitive viruses. We, then, examined the antiviral susceptibility of 57 pairs of influenza viruses isolated from patients pre- and post-administration of favipiravir in phase 3 clinical trials. We found that there were no viruses with statistically significant reduced susceptibility to favipiravir or NA inhibitors, although two of 20 paired A(H1N1)pdm09, one of 17 paired A(H3N2) and one of 20 paired B viruses possessed amino acid substitutions in the RNA-dependent RNA polymerase subunits, PB1, PB2 and PA, after favipiravir administration. This is the first report on the antiviral susceptibility of influenza viruses isolated from patients after favipiravir treatment. PMID:27321665

  15. Isolation and Metagenomic Identification of Avian Leukosis Virus Associated with Mortality in Broiler Chicken

    PubMed Central

    2016-01-01

    Avian leukosis virus (ALV) belongs to the family Retroviridae and causes considerable economic losses to the poultry industry. Following an outbreak associated with high mortality in a broiler flock in northern part of Malaysia, kidney tissues from affected chickens were submitted for virus isolation and identification in chicken embryonated egg and MDCK cells. Evidence of virus growth was indicated by haemorrhage and embryo mortality in egg culture. While viral growth in cell culture was evidenced by the development of cytopathic effects. The isolated virus was purified by sucrose gradient and identified using negative staining transmission electron microscopy. Further confirmation was achieved through next-generation sequencing and nucleotide sequence homology search. Analysis of the viral sequences using the NCBI BLAST tool revealed 99-100% sequence homology with exogenous ALV viral envelope protein. Phylogenetic analysis based on partial envelope sequences showed the Malaysian isolate clustered with Taiwanese and Japanese ALV strains, which were closer to ALV subgroup J, ALV subgroup E, and recombinant A/E isolates. Based on these findings, ALV was concluded to be associated with the present outbreak. It was recommended that further studies should be conducted on the molecular epidemiology and pathogenicity of the identified virus isolate. PMID:27597865

  16. Genetic and phylogenetic analysis of glycoprotein of rabies virus isolated from several species in Brazil.

    PubMed

    Sato, Go; Itou, Takuya; Shoji, Youko; Miura, Yasuo; Mikami, Takeshi; Ito, Mikako; Kurane, Ichiro; Samara, Samir I; Carvalho, Adolorata A B; Nociti, Darci P; Ito, Fumio H; Sakai, Takeo

    2004-07-01

    Genetic and phylogenetic analyses of the region containing the glycoprotein (G) gene, which is related to pathogenicity and antigenicity, and the G-L intergenic region were carried out in 14 Brazilian rabies virus isolates. The isolates were classified as dog-related rabies virus (DRRV) or vampire bat-related rabies virus (VRRV), by nucleoprotein (N) analysis. The nucleotide and amino acid (AA) homologies of the area containing the G protein gene and G-L intergenic region were generally lower than those of the ectodomain. In both regions, nucleotide and deduced AA homologies were lower among VRRVs than among DRRVs. There were AA differences between DRRV and VRRV at 3 antigenic sites and epitopes (IIa, WB+ and III), suggesting that DRRV and VRRV can be distinguished by differences of antigenicity. In a comparison of phylogenetic trees between the ectodomain and the area containing the G protein gene and G-L intergenic region, the branching patterns of the chiropteran and carnivoran rabies virus groups differed, whereas there were clear similarities in patterns within the DRRV and VRRV groups. Additionally, the VRRV isolates were more closely related to chiropteran strains isolated from Latin America than to Brazilian DRRV. These results indicate that Brazilian rabies virus isolates can be classified as DRRV or VRRV by analysis of the G gene and the G-L intergenic region, as well as by N gene analysis. PMID:15297743

  17. Porcine reproductive and respiratory syndrome virus: genetic diversity of recent British isolates.

    PubMed

    Frossard, Jean-Pierre; Hughes, Gareth J; Westcott, David G; Naidu, Brindha; Williamson, Susanna; Woodger, Nicholas G A; Steinbach, Falko; Drew, Trevor W

    2013-03-23

    Porcine reproductive and respiratory syndrome (PRRS) continues to be a significant problem for European pig producers, contributing to porcine respiratory disease complex, neonatal piglet mortality, infertility and occasional abortion storms. PRRS virus (PRRSV), a member of the arterivirus family with two defined major genotypes, has been shown to be quite genetically diverse. In the present study, genetic analysis of multiple gene regions of over 100 viruses isolated in Britain between 2003 and 2007 revealed that the diversity of British strains is now far greater than during the early 1990s. All isolates belong to genotype 1 (European). While some recent isolates are still very similar to early isolates, a wide range of more diverse viruses is now also circulating. Interestingly, some isolates were found to be very similar to a modified-live vaccine strain, and it is suggested that use of the vaccine has affected the evolution pattern of PRRS virus strains in Britain. Evidence of deletions in one viral gene, ORF3, and of genome recombination was also seen. A molecular clock model using the ORF7 sequences estimates the rate of substitution as 3.8 × 10(-3) per site per year, thereby dating the most recent common ancestor of all British viruses to 1991, coincident with the first outbreak of disease. Our findings therefore have implications for both the diagnostic and prophylactic methods currently being used, which are discussed. PMID:23218831

  18. Isolation and Metagenomic Identification of Avian Leukosis Virus Associated with Mortality in Broiler Chicken.

    PubMed

    Bande, Faruku; Arshad, Siti Suri; Omar, Abdul Rahman

    2016-01-01

    Avian leukosis virus (ALV) belongs to the family Retroviridae and causes considerable economic losses to the poultry industry. Following an outbreak associated with high mortality in a broiler flock in northern part of Malaysia, kidney tissues from affected chickens were submitted for virus isolation and identification in chicken embryonated egg and MDCK cells. Evidence of virus growth was indicated by haemorrhage and embryo mortality in egg culture. While viral growth in cell culture was evidenced by the development of cytopathic effects. The isolated virus was purified by sucrose gradient and identified using negative staining transmission electron microscopy. Further confirmation was achieved through next-generation sequencing and nucleotide sequence homology search. Analysis of the viral sequences using the NCBI BLAST tool revealed 99-100% sequence homology with exogenous ALV viral envelope protein. Phylogenetic analysis based on partial envelope sequences showed the Malaysian isolate clustered with Taiwanese and Japanese ALV strains, which were closer to ALV subgroup J, ALV subgroup E, and recombinant A/E isolates. Based on these findings, ALV was concluded to be associated with the present outbreak. It was recommended that further studies should be conducted on the molecular epidemiology and pathogenicity of the identified virus isolate. PMID:27597865

  19. Genetic diversity of Japanese encephalitis virus isolates obtained from the Indonesian archipelago between 1974 and 1987.

    PubMed

    Schuh, Amy J; Guzman, Hilda; Tesh, Robert B; Barrett, Alan D T

    2013-07-01

    Five genotypes (GI-V) of Japanese encephalitis virus (JEV) have been identified, all of which have distinct geographical distributions and epidemiologies. It is thought that JEV originated in the Indonesia-Malaysia region from an ancestral virus. From that ancestral virus GV diverged, followed by GIV, GIII, GII, and GI. Genotype IV appears to be confined to the Indonesia-Malaysia region, as GIV has been isolated in Indonesia from mosquitoes only, while GV has been isolated on three occasions only from a human in Malaysia and mosquitoes in China and South Korea. In contrast, GI-III viruses have been isolated throughout Asia and Australasia from a variety of hosts. Prior to this study only 13 JEV isolates collected from the Indonesian archipelago had been studied genetically. Therefore the sequences of the envelope (E) gene of 24 additional Indonesian JEV isolates, collected throughout the archipelago between 1974 and 1987, were determined and a series of molecular adaptation analyses were performed. Phylogenetic analysis indicated that over a 14-year time span three genotypes of JEV circulated throughout Indonesia, and a statistically significant association between the year of virus collection and genotype was revealed: isolates collected between 1974 and 1980 belonged to GII, isolates collected between 1980 and 1981 belonged to GIV, and isolates collected in 1987 belonged to GIII. Interestingly, three of the GII Indonesian isolates grouped with an isolate that was collected during the JE outbreak that occurred in Australia in 1995, two of the GIII Indonesian isolates were closely related to a Japanese isolate collected 40 years previously, and two Javanese GIV isolates possessed six amino acid substitutions within the E protein when compared to a previously sequenced GIV isolate collected in Flores. Several amino acids within the E protein of the Indonesian isolates were found to be under directional evolution and/or co-evolution. Conceivably, the tropical climate

  20. Genetic Diversity of Japanese Encephalitis Virus Isolates Obtained from the Indonesian Archipelago Between 1974 and 1987

    PubMed Central

    Schuh, Amy J.; Guzman, Hilda; Tesh, Robert B.

    2013-01-01

    Abstract Five genotypes (GI–V) of Japanese encephalitis virus (JEV) have been identified, all of which have distinct geographical distributions and epidemiologies. It is thought that JEV originated in the Indonesia-Malaysia region from an ancestral virus. From that ancestral virus GV diverged, followed by GIV, GIII, GII, and GI. Genotype IV appears to be confined to the Indonesia-Malaysia region, as GIV has been isolated in Indonesia from mosquitoes only, while GV has been isolated on three occasions only from a human in Malaysia and mosquitoes in China and South Korea. In contrast, GI–III viruses have been isolated throughout Asia and Australasia from a variety of hosts. Prior to this study only 13 JEV isolates collected from the Indonesian archipelago had been studied genetically. Therefore the sequences of the envelope (E) gene of 24 additional Indonesian JEV isolates, collected throughout the archipelago between 1974 and 1987, were determined and a series of molecular adaptation analyses were performed. Phylogenetic analysis indicated that over a 14-year time span three genotypes of JEV circulated throughout Indonesia, and a statistically significant association between the year of virus collection and genotype was revealed: isolates collected between 1974 and 1980 belonged to GII, isolates collected between 1980 and 1981 belonged to GIV, and isolates collected in 1987 belonged to GIII. Interestingly, three of the GII Indonesian isolates grouped with an isolate that was collected during the JE outbreak that occurred in Australia in 1995, two of the GIII Indonesian isolates were closely related to a Japanese isolate collected 40 years previously, and two Javanese GIV isolates possessed six amino acid substitutions within the E protein when compared to a previously sequenced GIV isolate collected in Flores. Several amino acids within the E protein of the Indonesian isolates were found to be under directional evolution and/or co-evolution. Conceivably, the

  1. 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

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

    1994-01-01

    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

  2. Isolation and partial characterization of a novel virus from different carp species suffering gill necrosis - ultrastructure and morphogenesis.

    PubMed

    Granzow, H; Fichtner, D; Schütze, H; Lenk, M; Dresenkamp, B; Nieper, H; Mettenleiter, T C

    2014-06-01

    Two isolates of a novel enveloped RNA virus were obtained from carp and koi carp with gill necrosis. Both isolates behaved identically and could be propagated in different cyprinid cell lines forming large syncytia. The virus was sensitive to lipid solvents and neither exhibited haemadsorption/haemagglutination nor reverse transcriptase activity. Mature virus particles displayed a spherical shape with diameter of 100-350 nm after negative staining and 100-300 nm in ultrathin sections, covered by short projections of 8-10 nm in length. Maturation of virus progeny was shown to occur by budding and envelopment of the filamentous helical nucleocapsids at the cell surface. A detailed comparison of ultrastructure and morphogenesis of the novel virus isolates with selected arena-, ortho- and paramyxoviruses as possible candidates for evaluation of taxonomic classification yielded no consistency in all phenotypic features. Thus, on the basis of ultrastructure the novel virus isolates could not be assigned unequivocally to any established virus family. PMID:23865968

  3. Variability of geographically distinct isolates of maize rayado fino virus in Latin America.

    PubMed

    Hammond, R W; Kogel, R; Ramirez, P

    1997-12-01

    We have examined the molecular epidemiology of the leafhopper-borne maize rayado fino virus (MRFV) in Latin America. The coat protein gene and 3' non-translated region of 14 isolates of MRFV collected from Latin America and the United States were sequenced and phylogenetic relationships examined. The nucleotide sequence revealed remarkable conservation, with a sequence similarity of 88-99%. Phylogenetic analysis of sequence data obtained from a 633 bp fragment showed that MRFV has diverged into three main clusters, i.e. the geographically distinct northern and southern isolates and the Colombian isolates. Significant differences between the isolates collected from Colombia, previously named maize rayado colombiana virus, based upon differences in symptomatology and serological relationships to MRFV, and the other MRFV isolates, provides additional evidence supporting its designation as a unique strain of MRFV. PMID:9400964

  4. Complete genome sequence of two genotype III Japanese encephalitis virus isolates from West Bengal, India

    PubMed Central

    Taraphdar, Debjani; Chatterjee, Shyamalendu

    2015-01-01

    Background & objectives: Japanese encephalitis (JE), caused by a mosquito-borne virus JE virus (JEV), is a serious health problem in West Bengal, India. In this study, we report the complete genome sequence of two JEV isolates from West Bengal. The amino acid and nucleotide sequence homology was compared with other Indian strains. Methods: Two JEV isolates (IND-WB-JE1 and IND-WB-JE2) obtained in 2008 and 2010, respectively, from two districts of the State of West Bengal, respectively were analyzed for genetic variations by sequencing the 10934 bp whole genome of the virus. Of these two districts, one was covered under JE vaccination programme in 2007. Results: Phylogenetic analysis showed that both the isolates belonged to the genotype III. A total of 16 mutations were identified in the two isolates studied with respect to Vellore P20778 strain. One unique mutation A3215S was only found in IND-WB-JE2 isolate, but not in the isolate IND-WB-JE1. These two isolates showed maximum homology with P20778 strain of India. Interpretation & conclusions: This study reports on complete gene based phylogenetic analysis of JEV isolates from the State of West Bengal. It was evident from the results that JEV was still under circulation in both vaccine covered and not covered districts of West Bengal. PMID:26261169

  5. Genome Sequence of Foot-and-Mouth Disease Virus Serotype O Isolated from Morocco in 2015

    PubMed Central

    Wadsworth, J.; Gray, A.; Abouchoaib, N.; King, D. P.; Knowles, N. J.

    2016-01-01

    The genome of a virus isolated from an outbreak of foot-and-mouth disease (FMD) in Morocco in 2015 is described here. This virus is classified as lineage Ind-2001d within serotype O, topotype ME-SA (Middle East-South Asia). This lineage is endemic on the Indian subcontinent but has caused outbreaks in the Middle East and North Africa since 2013. PMID:27103736

  6. Genome Sequence of Foot-and-Mouth Disease Virus Serotype O Isolated from Morocco in 2015.

    PubMed

    Bachanek-Bankowska, K; Wadsworth, J; Gray, A; Abouchoaib, N; King, D P; Knowles, N J

    2016-01-01

    The genome of a virus isolated from an outbreak of foot-and-mouth disease (FMD) in Morocco in 2015 is described here. This virus is classified as lineage Ind-2001d within serotype O, topotype ME-SA (Middle East-South Asia). This lineage is endemic on the Indian subcontinent but has caused outbreaks in the Middle East and North Africa since 2013. PMID:27103736

  7. Isolation and characterization of a Korean porcine epidemic diarrhea virus strain KNU-141112.

    PubMed

    Lee, Sunhee; Kim, Youngnam; Lee, Changhee

    2015-10-01

    Severe outbreaks of porcine epidemic diarrhea virus (PEDV) have re-emerged in Korea and rapidly swept across the country, causing tremendous economic losses to producers and customers. Despite the availability of PEDV vaccines in the domestic market, the disease continues to plague the Korean pork industry, raising issues regarding their protective efficacy and new vaccine development. Therefore, PEDV isolation in cell culture is urgently needed to develop efficacious vaccines and diagnostic assays and to conduct further studies on the virus biology. In the present study, one Korean PEDV strain, KOR/KNU-141112/2014, was successfully isolated and serially propagated in Vero cells for over 30 passages. The in vitro and in vivo characteristics of the Korean PEDV isolate were investigated. Virus production in cell culture was confirmed by cytopathology, immunofluorescence, and real-time RT-PCR. The infectious virus titers of the viruses during the first 30 passages ranged from 10(5.1) to 10(8.2) TCID50 per ml. The inactivated KNU-141112 virus was found to mediate potent neutralizing antibody responses in immunized guinea pigs. Animal studies showed that KNU-141112 virus causes severe diarrhea and vomiting, fecal shedding, and acute atrophic enteritis, indicating that strain KNU-141112 is highly enteropathogenic in the natural host. In addition, the entire genomes or complete S genes of KNU-141112 viruses at selected cell culture passages were sequenced to assess the genetic stability and relatedness. Our genomic analyses indicated that the Korean isolate KNU-141112 is genetically stable during the first 30 passages in cell culture and is grouped within subgroup G2b together with the recent re-emergent Korean strains. PMID:26196056

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

    PubMed

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

    2014-12-01

    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

  9. Phylogenetic analysis of dengue virus types 1 and 3 isolated in Jakarta, Indonesia in 1988.

    PubMed

    Sjatha, Fithriyah; Takizawa, Yamato; Yamanaka, Atsushi; Konishi, Eiji

    2012-12-01

    Dengue viruses are mosquito-borne viruses that cause dengue fever and dengue hemorrhagic fever, both of which are globally important diseases. These viruses have evolved in a transmission cycle between human hosts and mosquito vectors in various tropical and subtropical environments. We previously isolated three strains of dengue type 1 virus (DENV1) and 14 strains of dengue type 3 virus (DENV3) during an outbreak of dengue fever and dengue hemorrhagic fever in Jakarta, Indonesia in 1988. Here, we compared the nucleotide sequences of the entire envelope protein-coding region among these strains. The isolates were 97.6-100% identical for DENV1 and 98.8-100% identical for DENV3. All DENV1 isolates were included in two different clades of genotype IV and all DENV3 isolates were included in a single clade of genotype I. For DENV1, three Yap Island strains isolated in 2004 were the only strains closely related to the present isolates; the recently circulated Indonesian strains were in different clades. Molecular clock analyses estimated that ancestors of the genotype IV strains of DENV1 have been indigenous in Indonesia since 1948. We predict that they diverged frequently around 1967 and that their offspring distributed to Southeast Asia, the Western Pacific, and Africa. For DENV3, the clade containing all the present isolates also contained strains isolated from other Indonesian regions and other countries including Malaysia, Singapore, China, and East Timor from 1985-2010. Molecular clock analyses estimated that the common ancestor of the genotype I strains of DENV3 emerged in Indonesia around 1967 and diverged frequently until 1980, and that their offspring distributed mainly in Southeast Asia. The first dengue outbreak in 1968 and subsequent outbreaks in Indonesia might have influenced the divergence and distribution of the DENV1 genotype IV strains and the DENV3 genotype I strains in many countries. PMID:22959957

  10. Antigenic and genetic analysis of a recently isolated H1N1 swine influenza virus.

    PubMed

    Olsen, C W; McGregor, M W; Cooley, A J; Schantz, B; Hotze, B; Hinshaw, V S

    1993-10-01

    Hemagglutinins (HA) of H1N1 swine influenza viruses isolated in the United States have remained antigenically and genetically conserved for many years. In contrast to such conservation, the HA of A/Swine/Nebraska/1/92 (Sw/Neb) could readily be distinguished from those of contemporary porcine viruses. Twenty-eight amino acid mutations differentiated the HA of Sw/Neb and A/Swine/Indiana/1726/88, the most recent H1N1 swine influenza virus for which HA sequence data were available. Among these differences were mutations at potential asparagine-linked glycosylation sites and charge changes at many residues. The Sw/Neb virus also could be differentiated from other swine influenza viruses in hemagglutination-inhibition assays with monoclonal antibodies to recent H1 swine HA. Nonetheless, overall sequence analysis of the HA and the nucleoprotein genes of Sw/Neb indicated that this virus was more closely related genetically to classic H1N1 swine influenza viruses than to H1N1 avian or human viruses. Infection of swine with Sw/Neb under experimental conditions induced clinical signs and lesions typical of swine influenza. However, affected swine in the field had high, persistent fevers, but relatively mild signs of respiratory tract disease. This study indicated that an antigenically and genetically novel variant of swine influenza virus was detected in the United States. PMID:8250388

  11. Opium poppy mosaic virus, a new umbravirus isolated from Papaver somniferum in New Zealand.

    PubMed

    Tang, Joe; Lebas, Bénédicte; Liefting, Lia; Veerakone, Stella; Wei, Ting; Ward, Lisa

    2016-01-01

    A novel virus, tentatively named "opium poppy mosaic virus" (OPMV), was isolated from Papaver somniferum (opium poppy) with leaf mosaic and mottling symptoms in Auckland, New Zealand, in 2006. The virus was mechanically transmitted to herbaceous plants of several species, in which it induced local and/or systemic symptoms. No virus particles were observed by electron microscopy in the diseased P. somniferum or any of the symptomatic herbaceous plants. The complete genomic sequence of 4230 nucleotides contains four open reading frames (ORF) and is most closely related (59.3 %) to tobacco bushy top virus, a member of the genus Umbravirus. These data suggest that OPMV is a new umbravirus. PMID:26514844

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

    PubMed Central

    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.; Abrahão, Jônatas S.

    2014-01-01

    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

  13. Isolation and Complete Genome Sequencing of Bluetongue Virus Serotype 12 from India.

    PubMed

    Rao, P P; Reddy, Y V; Hegde, N R

    2015-10-01

    Bluetongue virus (BTV) causes disease mainly in sheep, but can be transmitted via other domestic and wild ruminants, resulting in pecuniary burden and trade restrictions. Segmented genome with the possibility of reassortment, existence of 26 serotypes, geographical restriction in the distribution of many of the serotypes, use of live attenuated vaccines and the lack of complete sequences of viruses isolated from several parts of the globe have complicated our understanding of the origin, movement and distribution of BTV. Recent efforts in genome sequencing of several strains have helped in better comprehending BTV epidemiology. In an effort to contribute to the genetic epidemiology of BTV in India, we report the isolation and complete genome sequencing of a BTV serotype 12 virus (designated NMO1). This is the first BTV-12 isolated from India and the second BTV-12 to be sequenced worldwide. The analysis of sequences of this virus suggests that NMO1 derived its segments from viruses belonging to western topotype viruses, as well as those from South-East Asia and India. The results have implications for understanding the origin, emergence/re-emergence and movement of BTV as well as for the development of vaccines and diagnostics based on robust epidemiological data. PMID:24330644

  14. Resistance breaking tomato spotted wilt virus isolates on resistant pepper varieties in Italy.

    PubMed

    Crescenzi, A; Viggiano, A; Fanigliulo, A

    2013-01-01

    In spring 2012, resistance breaking (RB) isolates of tomato spotted wilt virus (TSWV) that overcome the resistance conferred by the Tsw gene in different pepper hybrids have been recovered in different locations in southern Italy (Campania and Apulia regions) in protected cultivation, about one month after transplant. The percentage of symptomatic plants was 5-10% and, only in particular cases of advanced stage of cultivation, it reached 30-50% at the end of cycle. All TSWV isolates induced similar systemic symptoms in all resistant infected pepper hybrids: yellowing or browning of apical leaves, which later become necrotic, long necrotic streakson stems, extending to the terminal shoots, complete necrosis of younger fruits and large necrotic streaks and spots on fruits formed after infection. On ripe fruits, yellow spots with concentric rings or necrotic streaks could be observed. Leaf extracts of these samples were tested in ELISA for the detection of TSWV, Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV), Tomato mosaic virus (ToMV), Impatiens necrotic spot virus (INSV), Potato virus Y (PVY), Alfalfa mosaic virus (AMV), Pepper mild mottle virus (PMMoV) and Pepper Mottle Virus (PepMoV). Only TSWV was detected in all the field samples tested. The correspondent virus isolates were inoculated mechanically and by Frankliniella occidentalis on to a set of different pepper and tomato hybrids, as well as on some herbaceous test plants, in order to investigate for their ability to overcome the resistance genes Tsw and Sw5, respectively. Tomato hybrids carrying the Sw5 gene were uninfected by all RB isolates, whereas all resistant pepper hybrids became systemically infected. RB isolates did not differ noticeably in transmission efficiency when they were tested with the thrips F. occidentalis. Obtained results demonstrate that evolved strains of TSWV have emerged, that they are able to overcome the Tsw resistance gene in pepper plants experimentally inoculated both

  15. Thermostability of subpopulations of H2N3 influenza virus isolates from mallard ducks.

    PubMed

    Negovetich, Nicholas J; Webster, Robert G

    2010-09-01

    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 degrees C and 55 degrees C, and infectivity titers were calculated at different time points. Based on the rate of infectivity inactivation at 37 degrees 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 degrees 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

  16. Sequence analysis and genetic diversity of five new Indian isolates of cucumber mosaic virus.

    PubMed

    Kumar, S; Gautam, K K; Raj, S K

    2015-12-01

    Cucumber mosaic virus (CMV) is an important virus since it causes severe losses to many economically important crops worldwide. Five new isolates of CMV were isolated from naturally infected Hippeastrum hybridum, Dahlia pinnata, Hemerocallis fulva, Acorus calamus and Typhonium trilobatum plants, all exhibiting severe leaf mosaic symptoms. For molecular identification and sequence analyses, the complete coat protein (CP) gene of these isolates was amplified by RT-PCR. The resulting amplicons were cloned and sequenced and isolates were designated as HH (KP698590), DP (JF682239), HF (KP698589), AC (KP698588) and TT (JX570732). For study of genetic diversity among these isolates, the sequence data were analysed by BLASTn, multiple alignment and generating phylogenetic trees along with the respective sequences of other CMV isolates available in GenBank Database were done. The isolates under study showed 82-99% sequence diversity among them at nucleotide and amino acid levels; however they showed close relationships with CMV isolates of subgroup IB. In alignment analysis of amino acid sequences of HH and AC isolates, we have found fifteen and twelve unique substitutions, compared to HF, DP and TT isolates, suggesting the cause of high genetic diversity. PMID:26666188

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

    SciTech Connect

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

    1981-04-01

    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.

  18. Isolation and Characterization of Influenza C Viruses in the Philippines and Japan

    PubMed Central

    Odagiri, Takashi; Matsuzaki, Yoko; Okamoto, Michiko; Suzuki, Akira; Saito, Mariko; Tamaki, Raita; Lupisan, Socorro P.; Sombrero, Lydia T.; Hongo, Seiji

    2014-01-01

    From November 2009 to December 2013 in the Philippines, 15 influenza C viruses were isolated, using MDCK cells, from specimens obtained from children with severe pneumonia and influenza-like illness (ILI). This is the first report of influenza C virus isolation in the Philippines. In addition, from January 2008 to December 2013, 7 influenza C viruses were isolated from specimens that were obtained from children with acute respiratory illness (ARI) in Sendai city, Japan. Antigenic analysis with monoclonal antibodies to the hemagglutinin-esterase (HE) glycoprotein showed that 19 strains (12 from the Philippines and 7 from Japan) were similar to the influenza C virus reference strain C/Sao Paulo/378/82 (SP82). Phylogenetic analysis of the HE gene showed that the strains from the Philippines and Japan formed distinct clusters within an SP82-related lineage. The clusters that included the Philippine and Japanese strains were shown to have diverged from a common ancestor around 1993. In addition, phylogenetic analysis of the internal genes showed that all strains isolated in the Philippines and Japan had emerged through reassortment events. The composition of the internal genes of the Philippine strains was different from that of the Japanese strains, although all strains were classified into an SP82-related lineage by HE gene sequence analysis. These observations suggest that the influenza C viruses analyzed here had emerged through different reassortment events; however, the time and place at which the reassortment events occurred were not determined. PMID:25552361

  19. [Phenotyte of epidemic influenza B virus strains isolated in different years].

    PubMed

    Larionova, N V; Kiseleva, I V; Isakova, I N; Litvinova, O M; Rudenko, L G

    2006-01-01

    The reproducing ability at elevated temperatures (non-ts phenotype) was examined for 38 influenza B virus strains isolated in different years in different countries. Out of the 7 strains isolated in 1940 to 1973, only one showed temperature-sensitivity of reproduction (a ts phenotype). In 1984 to 1988, the proportion of temperature-sensitive strains increased up to 55% (6 of 11). Since the late 1990s, the majority (90%) of the study influenza B viruses demonstrated a pronounced ts phenotype. Influenza B virus strains were also examined for their resistance to serum inhibitors. Prior to the divergence of influenza B viruses into two lines: B/Jamagata and B/Victoria, the epidemic viruses exhibited a high resistance to nonspecific inhibitors of normal equine serum. This property was also preserved in all study B/Victoria strains; however, 83% of the B/Jamagata viruses were inhibitor-sensitive. The present study has demonstrated the heterogenicity of epidemic influenza B viruses in temperature- and inhibitor-sensitivity. PMID:17087064

  20. Characterisation of the welsh onion isolate of Shallot yellow stripe virus from China.

    PubMed

    Chen, J; Wei, C-B; Zheng, H-Y; Shi, Y-H; Adams, M J; Lin, L; Zhang, Q-Y; Wang, S-J; Chen, J-P

    2005-10-01

    The host range and nucleotide sequence of shallot yellow stripe virus (SYSV) from welsh onion in Shandong province, China is described. Of the plants tested, only shallot and welsh onion became infected but most shallot plants were symptomless. The complete sequence of one isolate (10429 nt) and the 3'-terminal 3540 nts of a second isolate were determined. They had c. 90% nt identity to one another and to published (partial) sequences of SYSV. SYSV was most closely related to onion yellow dwarf virus (OYDV) and resembled it in having a much larger P3 protein than other species in the genus. PMID:15968472

  1. Molecular Characteristics of H6N6 Influenza Virus Isolated from Pigeons in Guangxi, Southern China

    PubMed Central

    Li, Meng; Xie, Zhiqin; Luo, Sisi; Xie, Liji; Huang, Li; Deng, Xianwen; Huang, Jiaoling; Zhang, Yanfang; Zeng, Tingting; Wang, Sheng

    2015-01-01

    Here, we report the complete genome sequence of an H6N6 avian influenza virus (AIV) isolated from a pigeon in Guangxi, southern China, in 2014. The eight RNA segment genes shared a high nucleotide identity (97 to 99%) with H6N6 subtypes of AIV isolated from ducks in the regions around Guangxi Province. The finding of this study will help us understand the ecology and molecular characteristics of H6 avian influenza virus in wild birds in southern China. PMID:26634763

  2. Genome organization and host range of a Brazilian isolate of johnsongrass mosaic virus.

    PubMed

    Camelo-García, Viviana Marcela; da Silva Andrade, Sónia Cristina; Geering, Andrew D W; Kitajima, Elliot Watanabe; Rezende, Jorge A M

    2016-05-01

    This work reports the complete genome sequence, production of a polyclonal antiserum, and host range of a Brazilian strain of johnsongrass mosaic virus (JGMV) found infecting Panicum maximum in the state of São Paulo, Brazil. The complete genome sequence of this potyvirus, comprising 9874 nucleotides, showed 82 % amino acid sequence identity in the polyprotein to that of an isolate of JGMV from Australia. The experimental host range of this virus included mainly fodder species. Cultivated species such as rice, oats, sugarcane, rye, corn and wheat were not infected, suggesting that current isolates of this potyvirus do not represent a threat to these crops in Brazil. PMID:26869279

  3. Complete Genome Sequence of Zika Virus Isolated in Mexico, 2016

    PubMed Central

    Peña-Alonso, Rocío; Mendieta-Condado, Edgar; Garcés-Ayala, Fabiola; González-Durán, Elizabeth; Escobar-Escamilla, Noé; Vázquez-Pichardo, Mauricio; Torres-Rodríguez, María de la Luz; Núñez-León, Alma; Torres-Longoria, Belem; López-Martínez, Irma; Ruíz-Matus, Cuitláhuac; Kuri-Morales, Pablo

    2016-01-01

    Zika virus belongs to the genus Flavivirus, and its spread remains an international public health emergency. In this report, we describe the obtainment and molecular characterization of a complete viral genome through the direct metagenomic analysis from saliva from an autochthonous transmission case in Mexico. PMID:27491989

  4. Diversity of Papaya ringspot virus isolates in Puerto Rico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Papaya ringspot virus (PRSV) devastates papaya production worldwide. In Puerto Rico, papaya fields can be completely infected with PRSV within a year of planting. Information about the diversity of the Puerto Rican PRSV population is relevant in order to establish a control strategy in the island. T...

  5. Complete Genome Sequence of Zika Virus Isolated in Mexico, 2016.

    PubMed

    Díaz-Quiñonez, José Alberto; Peña-Alonso, Rocío; Mendieta-Condado, Edgar; Garcés-Ayala, Fabiola; González-Durán, Elizabeth; Escobar-Escamilla, Noé; Vázquez-Pichardo, Mauricio; Torres-Rodríguez, María de la Luz; Núñez-León, Alma; Torres-Longoria, Belem; López-Martínez, Irma; Ruíz-Matus, Cuitláhuac; Kuri-Morales, Pablo; Ramírez-González, José Ernesto

    2016-01-01

    Zika virus belongs to the genus Flavivirus, and its spread remains an international public health emergency. In this report, we describe the obtainment and molecular characterization of a complete viral genome through the direct metagenomic analysis from saliva from an autochthonous transmission case in Mexico. PMID:27491989

  6. Pathogenesis and Transmission of Feral Swine Pseudorabies Virus Isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction. Aujesky’s Disease or pseudorabies, is one of the oldest recognized swine diseases. It is caused by pseudorabies virus (PRV), an alpha-herpesvirus that can induce respiratory disease, reproductive failure, and affect the central nervous system. PRV vaccines, in conjunction with serologi...

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

    USGS Publications Warehouse

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

    2005-01-01

    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.

  8. Genomic Characterizations of a Newcastle Disease Virus Isolated from Ducks in Live Bird Markets in China

    PubMed Central

    Zhang, Yi; Zheng, Dongxia; Zhao, Yunling; Castellan, David; Liu, Hualei; Wang, Zhiliang

    2016-01-01

    One class I Newcastle disease virus (NDV), designated as duck/Guangxi/1261/2015 (GX1261), was isolated from asymptomatic ducks in live bird markets (LBM) from southern China during the national active surveillance for NDVs in 2015. The complete genome length of GX1261 isolate was 15,198 nucleotides with the gene order of 3’-NP-P-M-F-HN-L-5’. The motif at the cleavage site of F protein was 112ERQER/L117, which was typical of low virulence NDV. Several mutations were identified in the functional domains of F and HN proteins, including fusion peptide, heptad repeat region, transmembrane domains and neutralizing epitopes. Phylogenetic analysis based on the complete F gene revealed that the isolate was clustered into sub-genotype 1c in class I, and showed a high level of similarity with the strains isolated from waterfowl in the United States of America. This is the first report of this kind of virus in the mainland of China. These results demonstrated that GX1261-like viruses might exist in asymptomatic waterfowl, and remain undetected or unidentified. Thus, more investigation needs to be done in order to identify the source of the virus. This study revealed the genetic and phylogenetic characteristics of GX1261 isolate and could help us to better understand the epidemiological context of class I NDV in China. PMID:27391305

  9. Genomic Characterizations of a Newcastle Disease Virus Isolated from Ducks in Live Bird Markets in China.

    PubMed

    Wang, Jingjing; Lv, Yan; Zhang, Yi; Zheng, Dongxia; Zhao, Yunling; Castellan, David; Liu, Hualei; Wang, Zhiliang

    2016-01-01

    One class I Newcastle disease virus (NDV), designated as duck/Guangxi/1261/2015 (GX1261), was isolated from asymptomatic ducks in live bird markets (LBM) from southern China during the national active surveillance for NDVs in 2015. The complete genome length of GX1261 isolate was 15,198 nucleotides with the gene order of 3'-NP-P-M-F-HN-L-5'. The motif at the cleavage site of F protein was 112ERQER/L117, which was typical of low virulence NDV. Several mutations were identified in the functional domains of F and HN proteins, including fusion peptide, heptad repeat region, transmembrane domains and neutralizing epitopes. Phylogenetic analysis based on the complete F gene revealed that the isolate was clustered into sub-genotype 1c in class I, and showed a high level of similarity with the strains isolated from waterfowl in the United States of America. This is the first report of this kind of virus in the mainland of China. These results demonstrated that GX1261-like viruses might exist in asymptomatic waterfowl, and remain undetected or unidentified. Thus, more investigation needs to be done in order to identify the source of the virus. This study revealed the genetic and phylogenetic characteristics of GX1261 isolate and could help us to better understand the epidemiological context of class I NDV in China. PMID:27391305

  10. Phenotypic and genotypic characterization of clinical isolates of herpes simplex virus resistant to aciclovir.

    PubMed

    Harris, Wendy; Collins, Peter; Fenton, Rob J; Snowden, Wendy; Sowa, Mike; Darby, Graham

    2003-06-01

    A panel of 10 clinical isolates of herpes simplex virus (HSV) deficient in the expression of thymidine kinase (TK) and phenotypically resistant to aciclovir was characterized. Sequence analysis revealed a variety of mutations in TK (nucleotide substitutions, insertions and deletions), most of which resulted in truncated TK polypeptides. In line with previous reports, the most common mutation was a single G insertion in the 'G-string' motif. One HSV-1 isolate and two HSV-2 isolates appeared to encode full-length polypeptides and, in each case, an amino acid substitution likely to be responsible for the phenotype was identified. Pathogenicity was determined using a zosteriform model of HSV infection in BALB/c mice. The majority of isolates appeared to show impaired growth at the inoculation site compared with wild-type virus. They also showed poor replication in the peripheral nervous system and little evidence of zosteriform spread. One exception was isolate 4, which had a double G insertion in the G-string but, nevertheless, exhibited zosteriform spread. These studies confirmed that TK-deficient viruses display a range of neurovirulence with respect to latency and zosteriform spread. These results are discussed in the light of previous experience with TK-deficient viruses. PMID:12771406

  11. Antigenic characterisation of virus isolates from vaccinated dogs dying of rabies.

    PubMed

    Aghomo, H O; Rupprecht, C E

    1990-11-01

    Four rabies virus isolates from dogs that succumbed to rabies infection in Nigeria within one year of anti-rabies vaccination were characterised by monoclonal antibodies (MAbs). The samples were screened for rabies and rabies-related viral antigens by the indirect fluorescent antibody test, performed with MAb 502-2, which recognises the nucleocapsid (NC) protein of all known Lyssaviruses and with MAb 422-5 which identifies African rabies-related viruses. All four canine virus isolates displayed positive fluorescence with MAb 502-2 and were negative with MAb 422-5. In the anti-NC MAb characterisation with a panel of 34 additional MAbs, all isolates displayed positive staining with 32 of the MAbs, were negative with MAb 102-27 and all displayed poor immunofluorescence with MAb 377-7. On the basis of reactivity with a panel of 40 anti-glycoprotein (G) MAbs the isolates were separated into four distinct viral subtypes. None of these canine isolates was identified as the common attenuated Flury LEP rabies strain used for domestic animal vaccination and none resembled other previously characterised rabies viruses from Nigeria. PMID:2288013

  12. Receptor specificity of subtype H1 influenza A viruses isolated from swine and humans in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The evolution of receptor specificity of classical swine influenza viruses leading to the 2009 H1N1 pandemic virus was analyzed in glycan microarrays. Classical influenza viruses from the alpha, beta, and gamma antigenic clusters isolated between 1945 and 2009 revealed a binding profile very simila...

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

    PubMed

    Farzadfar, Shirin; Pourrahim, Reza

    2013-10-01

    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

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

    PubMed Central

    2012-01-01

    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

  15. Comparison of the nucleotide sequences of wheat dwarf virus (WDV) isolates from Hungary and Ukraine.

    PubMed

    Tóbiás, Istvan; Shevchenko, Oleksiy; Kiss, Balázs; Bysov, Andriy; Snihur, Halina; Polischuk, Valery; Salánki, Katalin; Palkovics, László

    2011-01-01

    Wheat dwarf virus (WDV) is the most ubiquitous virus in cereals causing huge losses in both Hungary and Ukraine. The presence of barley-and wheat-adapted strains has been confirmed, suggesting that the barley strain is restricted to barley, while the wheat strain is present in both wheat and barley plants. Five WDV isolates from wheat plants sampled in Hungary and Ukraine were sequenced and compared with known WDV isolates from GenBank. Four WDV isolates belonged to the wheat strain. Our results indicate that WDV-Odessa is an isolate of special interest since it has originated from wheat, but belongs to the barley-adapted strain, providing novel data on WDV biology and raising issues of pathogen epidemiology. PMID:21905629

  16. Biological and molecular characterization of a highly divergent johnsongrass mosaic virus isolate from Pennisetum purpureum.

    PubMed

    Silva, Karina N; Melo, Fernando L; Orílio, Anelise F; Nagata, Tatsuya; Silva, Marilia S; Fernandes, Celso D; Fragoso, Rodrigo R; Dessaune, Suelen N; Resende, Renato O

    2016-07-01

    The complete genome sequence (9,865 nucleotides) of a highly divergent johnsongrass mosaic virus isolate (JGMV-CNPGL) was determined using Illumina sequencing. This isolate infected 10 genotypes of gramineous plants including maize. A comparative analysis of the complete genome showed 80 % nucleotide (nt) sequence identity (86 % amino acid (aa) sequence identity) to a johnsongrass mosaic virus isolate from Australia. The coat protein (CP) identity values, however, were lower than those for the whole genome (78 % and 80 % for nt and aa, respectively) and were close to the species demarcation values (77 % nt and 80 % aa). Unexpectedly, the amino-terminal portion of CP of JGMV-CNPGL showed only 38 % sequence identity to other JGMV isolates. The biological implications of this sequence divergence remain to be elucidated. PMID:27101070

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

    NASA Astrophysics Data System (ADS)

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

    1981-06-01

    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).

  18. FBJ osteosarcoma virus in tissue culture. III. Isolation and characterization of non-virus-producing FBJ-transformed cells.

    PubMed Central

    Levy, J A; Kazan, P L; Reilly, C A; Finkel, M P

    1978-01-01

    Hamster and rat cell lines have been established that have been transformed by FBJ murine sarcoma virus (FBJ-MuSV) but that do not produce virus. The hamster cell line originated from an osteosarcoma that appeared in a hamster inoculated at birth with an extract of a CFNo1 mouse FBJ-osteosarcoma. The rat cell line was obtained by transferring the FBJ-MuSV genome to normal rat kidney cells in the absence of the FBJ type C virus (FBJ-MuLV), which, usually in high concentration, accompanies the FBJ-MuSV. Both transformed hamster and rat cell lines contain the FBJ-MuSV genome, which can be rescued by ecotropic and xenotropic murine type C viruses. This rescued genome produces characteristic FBJ-MuSV foci in tissue culture and, in appropriate animal hosts, induces osteosarcomas typical of those induced by FBJ-MuSV. FBJ-MuSV was isolated originally from a parosteal osteosarcoma that occurred naturally in a mouse. Since there was no previous history of passage of the agent through any other animal species, these non-virus-producing hamster and rat cells transformed by FBJ-MuSV should be very helpful in molecular studies examining the origin of spontaneous sarcoma genomes in mice. PMID:206718

  19. Complete Genome Sequence of Genotype VI Newcastle Disease Viruses Isolated from Pigeons in Pakistan

    PubMed Central

    Wajid, Abdul; Rehmani, Shafqat Fatima; Sharma, Poonam; Goraichuk, Iryna V.; Dimitrov, Kiril M.

    2016-01-01

    Two complete genome sequences of Newcastle disease virus (NDV) are described here. Virulent isolates pigeon/Pakistan/Lahore/21A/2015 and pigeon/Pakistan/Lahore/25A/2015 were obtained from racing pigeons sampled in the Pakistani province of Punjab during 2015. Phylogenetic analysis of the fusion protein genes and complete genomes classified the isolates as members of NDV class II, genotype VI. PMID:27540069

  20. [The usefulness of the CLA-1 cell subline for the isolation of the dengue virus].

    PubMed

    Morier, L; Castillo, A; Rodríguez, R; Guadalupe Guzmán, M

    1995-01-01

    The live CLA-1 obtained by cloning from the cell live AP-61 (Aedes pseudocutellaris) was used to intend the isolation of dengue virus in 10 serum samples patients in acute phase and clinical diagnosis of dengue. Samples were parallelly inoculated in C6/36 and AP-61 as control systems. Of the 10 samples, 6 were positive in the 3 systems evidencing sensitivity and usefulness for the isolation of the cell subline CLA-1. PMID:9813481

  1. Complete Genome Sequence of Genotype VI Newcastle Disease Viruses Isolated from Pigeons in Pakistan.

    PubMed

    Wajid, Abdul; Rehmani, Shafqat Fatima; Sharma, Poonam; Goraichuk, Iryna V; Dimitrov, Kiril M; Afonso, Claudio L

    2016-01-01

    Two complete genome sequences of Newcastle disease virus (NDV) are described here. Virulent isolates pigeon/Pakistan/Lahore/21A/2015 and pigeon/Pakistan/Lahore/25A/2015 were obtained from racing pigeons sampled in the Pakistani province of Punjab during 2015. Phylogenetic analysis of the fusion protein genes and complete genomes classified the isolates as members of NDV class II, genotype VI. PMID:27540069

  2. Genomic characterization of influenza A (H7N9) viruses isolated in Shenzhen, Southern China, during the second epidemic wave.

    PubMed

    Fang, Shisong; Wang, Xin; Dong, Fangyuan; Jin, Tao; Liu, Guang; Lu, Xing; Peng, Bo; Wu, Weihua; Liu, Hui; Kong, Dongfeng; Tang, Xiujuan; Qin, Yanmin; Mei, Shujiang; Xie, Xu; He, Jianfan; Ma, Hanwu; Zhang, Renli; Cheng, Jinquan

    2016-08-01

    There were three epidemic waves of human infection with avian influenza A (H7N9) virus in 2013-2014. While many analyses of the genomic origin, evolution, and molecular characteristics of the influenza A (H7N9) virus have been performed using sequences from the first epidemic wave, genomic characterization of the virus from the second epidemic wave has been comparatively less reported. In this study, an in-depth analysis was performed with respect to the genomic characteristics of 11 H7N9 virus strains isolated from confirmed cases and four H7N9 virus strains isolated from environmental samples in Shenzhen during the second epidemic wave. Phylogenetic analysis demonstrated that six internal segments of the influenza A (H7N9) virus isolated from confirmed cases and environmental samples in Shenzhen were clustered into two different clades and that the origin of the influenza A (H7N9) virus isolated from confirmed cases in Shenzhen was different from that of viruses isolated during the first wave. In addition, H9N2 viruses, which were prevalent in southern China, played an important role in the reassortment of the influenza A (H7N9) virus isolated in Shenzhen. HA-R47K and -T122A, PB2-V139I, PB1-I397M, and NS1-T216P were the signature amino acids of the influenza A (H7N9) virus isolated from confirmed cases in Shenzhen. We found that the HA, NA, M, and PA genes of the A(H7N9) viruses underwent positive selection in the human population. Therefore, enhanced surveillance should be carried out to determine the origin and mode of transmission of the novel influenza A (H7N9) virus and to facilitate the formulation of effective policies for prevention and containment of a human infection epidemics. PMID:27169600

  3. Isolation and Genetic Characterization of Mangshi Virus: A Newly Discovered Seadornavirus of the Reoviridae Family Found in Yunnan Province, China

    PubMed Central

    Wang, Jinglin; Li, Huachun; He, Yuwen; Zhou, Yang; Meng, Jingxing; Zhu, Wuyang; Chen, Hongyu; Liao, Defang; Man, Yunping

    2015-01-01

    Background Seadornavirus is a genus of viruses in the family Reoviridae, which consists of Banna virus, Kadipiro virus, and Liao ning virus. Banna virus is considered a potential pathogen for zoonotic diseases. Here, we describe a newly discovered Seadornavirus isolated from mosquitos (Culex tritaeniorhynchus) in Yunnan Province, China, which is related to Banna virus, and referred to as Mangshi virus. Methods and Results The Mangshi virus was isolated by cell culture in Aedes albopictus C6/36 cells, in which it replicated and caused cytopathic effects, but not in mammalian BHK-21 or Vero cells. Polyacrylamide gel analysis revealed a genome consisting of 12 segments of double-stranded RNA, with a “6–4–2” pattern in which the migrating bands were different from those of the Banna virus. Complete genome sequencing was performed by full-length amplification of cDNAs. Sequence analysis showed that seven highly conserved nucleotides and three highly conserved nucleotides were present at the ends of the 5′- and 3′-UTRs in each of 12 genome segments. The amino acid identities of Mangshi virus shared with Balaton virus varied from 27.3% (VP11) to 72.3% (VP1) with Banna virus varying from 18.0% (VP11) to 63.9% (VP1). Phylogenetic analysis based on amino acid sequences demonstrated that Mangshi virus is a member of the genus Seadornavirus and is most closely related to, but distinct from, Balaton virus and Banna virus in the genus Seadornavirus of the family Reoviridae. Conclusion Mangshi virus isolated from mosquitoes (C. tritaeniorhynchus) was identified as a newly discovered virus in the genus Seadornavirus and is phylogenetically close to Banna virus, suggesting that there is genetic diversity of seadornaviruses in tropical and subtropical areas of Southeast Asia. PMID:26630378

  4. Molecular typing and phylogenetic analysis of classical swine fever virus isolates from Kerala, India.

    PubMed

    Bhaskar, Nimisha; Ravishankar, Chintu; Rajasekhar, R; Sumod, K; Sumithra, T G; John, Koshy; Mini, M; Ravindran, Reghu; Shaji, Shiju; Aishwarya, J

    2015-12-01

    Classical swine fever (CSF) is an economically important disease of pigs caused by CSF virus (CSFV) belonging to the genus Pestivirus within the family Flaviviridae. The disease is endemic in many countries including India. A comprehensive study was carried out to assess the type of CSFV circulating in the South Indian state of Kerala. During the period 2013-2014, clinical samples were collected from 19 suspected CSF outbreaks of domestic pigs in different districts of Kerala. The samples were tested using nested reverse transcription PCR (RT-PCR) targeting the E2 gene and RT-PCR for 5'UTR of the virus. Partial 5' UTR and E2 gene regions of six CSFV isolates were sequenced. Phylogenetic analysis revealed that all the CSFV isolates belonged to subgroup 2.2. The isolates showed close resemblance to the other CSFV isolates circulating in India. It was also observed that the CSFV viruses from Kannur district were distinct from those circulating in the other districts as evidenced by their divergence from other Kerala isolates in the phylogenetic tree. Close relationship was seen to the CSFV isolates from South East Asian countries. PMID:26645036

  5. Specific Insect-Virus Interactions Are Responsible for Variation in Competency of Different Thrips tabaci Isolines to Transmit Different Tomato Spotted Wilt Virus Isolates

    PubMed Central

    Jacobson, Alana L.; Kennedy, George G.

    2013-01-01

    Local adaptation between sympatric host and parasite populations driven by vector genetics appears to be a factor that influences dynamics of disease epidemics and evolution of insect-vectored viruses. Although T. tabaci is the primary vector of Tomato spotted wilt virus (TSWV) in some areas of the world, it is not an important vector of this economically important plant virus in many areas where it occurs. Previous studies suggest that genetic variation of thrips populations, virus isolates, or both are important factors underlying the localized importance of this species as a vector of TSWV. This study was undertaken to quantify variation in transmissibility of TSWV isolates by T. tabaci, in the ability of T. tabaci to transmit isolates of TSWV, and to examine the possibility that genetic interactions and local adaptation contribute to the localized nature of this species as a vector of TSWV. Isofemale lines of Thrips tabaci from multiple locations were tested for their ability to transmit multiple TSWV isolates collected at the same and different locations as the thrips. Results revealed that the probability of an isofemale line transmitting TSWV varied among virus isolates, and the probability of an isolate being transmitted varied among isofemale lines. These results indicate that the interaction of T. tabaci and TSWV isolate genetic determinants underlie successful transmission of TSWV by T. tabaci. Further analysis revealed sympatric vector-virus pairing resulted in higher transmission than allopatric pairing, which suggests that local adaptation is occurring between T. tabaci and TSWV isolates. PMID:23358707

  6. Ilheus Virus Isolation in the Pantanal, West-Central Brazil

    PubMed Central

    Pauvolid-Corrêa, Alex; Kenney, Joan L.; Couto-Lima, Dinair; Campos, Zilca M. S.; Nogueira, Rita M. R.; Brault, Aaron C.; Komar, Nicholas

    2013-01-01

    The wetlands of the Brazilian Pantanal host large concentrations of diverse wildlife species and hematophagous arthropods, conditions that favor the circulation of zoonotic arboviruses. A recent study from the Nhecolândia sub-region of Pantanal reported serological evidence of various flaviviruses, including West Nile virus and Ilheus virus (ILHV). According to the age of seropositive horses, at least three flaviviruses, including ILHV, circulated in the Brazilian Pantanal between 2005 and 2009. To extend this study, we collected 3,234 adult mosquitoes of 16 species during 2009 and 2010 in the same sub-region. Mosquito pool homogenates were assayed for infectious virus on C6/36 and Vero cell monolayers and also tested for flaviviral RNA by a group-specific real-time RT-PCR. One pool containing 50 non-engorged female specimens of Aedes scapularis tested positive for ILHV by culture and for ILHV RNA by real-time RT-PCR, indicating a minimum infection rate of 2.5 per 1000. Full-length genomic sequence exhibited 95% identity to the only full genome sequence available for ILHV. The present data confirm the circulation of ILHV in the Brazilian Pantanal. PMID:23875051

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

    PubMed Central

    2010-01-01

    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

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

    PubMed

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

    2010-01-01

    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

  9. Complete genome sequence of nine isolates of canna yellow streak virus reveals its relationship to the sugarcane mosaic virus (SCMV) subgroup of potyviruses.

    PubMed

    Chauhan, Ravendra P; Rajakaruna, Punsasi; Verchot, Jeanmarie

    2015-03-01

    Complete genome sequences were obtained from nine isolates of canna yellow streak virus (CaYSV). CaYSV belongs to the sugarcane mosaic virus (SCMV) subgroup of potyviruses with johnsongrass mosaic virus (JGMV) as its closest relative. Multiple sequence alignments showed a pattern of amino acid substitutions in the CP sequences, which enabled us to relate these isolates to South East Asian or European isolates. Biological characterization of CaYSV identified Nicotiana benthamiana, Chenopodium quinoa and Phaseolus vulgaris as experimental hosts. Given the popularity and global trade of cannas, a clear picture of the genetic diversity of CaYSV is critical to disease management. PMID:25567205

  10. Viruses of symbiotic Chlorella-like algae isolated from Paramecium bursaria and Hydra viridis

    PubMed Central

    Van Etten, James L.; Meints, Russel H.; Kuczmarski, Daniel; Burbank, Dwight E.; Lee, Kit

    1982-01-01

    We previously reported that isolation of symbiotic Chlorella-like algae from the Florida strain of Hydra viridis induced replication of a virus (designated HVCV-1) in the algae. We now report that isolation of symbiotic Chlorella-like algae from four other sources of green hydra and one source of the protozoan Paramecium bursaria also induced virus synthesis. Algae from one of these hydra contained a virus identical to HVCV-1 (based on its rate of sedimentation, buoyant density, reaction to HVCV-1 antiserum, and DNA restriction fragments) whereas algae from the other three hydra contained another similar, but distinct, virus (designated HVCV-2). The virus from the paramecium algae (designated PBCV-1) was distinct from both HVCV-1 and HVCV-2. The symbiotic algae in the hydra could also be distinguished ultrastructurally. Chloroplasts of both algae that produced HVCV-1 lacked a pyrenoid whereas chloroplasts of the other three symbiotic algae contained pyrenoids. Since all symbiotic eukaryotic algae we have examined have had virus, a potential viral role in symbiosis is suggested. Images PMID:16593198

  11. Syrian hamsters (Mesocricetus auratus) oronasally inoculated with a Nipah virus isolate from Bangladesh or Malaysia develop similar respiratory tract lesions.

    PubMed

    Baseler, L; de Wit, E; Scott, D P; Munster, V J; Feldmann, H

    2015-01-01

    Nipah virus is a paramyxovirus in the genus Henipavirus, which has caused outbreaks in humans in Malaysia, India, Singapore, and Bangladesh. Whereas the human cases in Malaysia were characterized mainly by neurological symptoms and a case fatality rate of ∼40%, cases in Bangladesh also exhibited respiratory disease and had a case fatality rate of ∼70%. Here, we compared the histopathologic changes in the respiratory tract of Syrian hamsters, a well-established small animal disease model for Nipah virus, inoculated oronasally with Nipah virus isolates from human cases in Malaysia and Bangladesh. The Nipah virus isolate from Bangladesh caused slightly more severe rhinitis and bronchointerstitial pneumonia 2 days after inoculation in Syrian hamsters. By day 4, differences in lesion severity could no longer be detected. Immunohistochemistry demonstrated Nipah virus antigen in the nasal cavity and pulmonary lesions; the amount of Nipah virus antigen present correlated with lesion severity. Immunohistochemistry indicated that both Nipah virus isolates exhibited endotheliotropism in small- and medium-caliber arteries and arterioles, but not in veins, in the lung. This correlated with the location of ephrin B2, the main receptor for Nipah virus, in the vasculature. In conclusion, Nipah virus isolates from outbreaks in Malaysia and Bangladesh caused a similar type and severity of respiratory tract lesions in Syrian hamsters, suggesting that the differences in human disease reported in the outbreaks in Malaysia and Bangladesh are unlikely to have been caused by intrinsic differences in these 2 virus isolates. PMID:25352203

  12. Genetic characterization of border disease virus (BDV) isolates from small ruminants in Italy.

    PubMed

    Giammarioli, Monica; Rossi, Elisabetta; Casciari, Cristina; Bazzucchi, Moira; Torresi, Claudia; Claudia, Torresi; De Mia, Gian Mario

    2015-04-01

    Border disease virus (BDV) belongs to the Pestivirus genus in the family Flaviviridae. Genetic analyses of pestiviruses that have been isolated from sheep and goat have led to the proposal that BDV isolates can be phylogenetically segregated into at least seven clusters, subtypes BDV-1 to BDV-7. In order to investigate the genetic heterogeneity of small ruminant pestivirus isolates in Italy, a selection of 5'-UTR sequences from isolates that were collected from clinical specimens between 2002 and 2014 was analysed. Phylogenetic reconstructions indicated that the BDV-positive samples clustered within the BDV-1, BDV-3, BDV-5, and BDV-7 groups. These results suggested high genetic diversity within the Italian BDV field isolates. The phylogenetic analysis indicated the first evidence of BDV-1 and BDV-5 circulation in Italy. The marked diversity of the pestivirus isolates might reflect the sheep trade with foreign countries. PMID:25636968

  13. Molecular characterization of banana bunchy top virus isolate from Sri Lanka and its genetic relationship with other isolates.

    PubMed

    Wickramaarachchi, W A R T; Shankarappa, K S; Rangaswamy, K T; Maruthi, M N; Rajapakse, R G A S; Ghosh, Saptarshi

    2016-06-01

    Bunchy top disease of banana caused by Banana bunchy top virus (BBTV, genus Babuvirus family Nanoviridae) is one of the most important constraints in production of banana in the different parts of the world. Six genomic DNA components of BBTV isolate from Kandy, Sri Lanka (BBTV-K) were amplified by polymerase chain reaction (PCR) with specific primers using total DNA extracted from banana tissues showing typical symptoms of bunchy top disease. The amplicons were of expected size of 1.0-1.1 kb, which were cloned and sequenced. Analysis of sequence data revealed the presence of six DNA components; DNA-R, DNA-U3, DNA-S, DNA-N, DNA-M and DNA-C for Sri Lanka isolate. Comparisons of sequence data of DNA components followed by the phylogenetic analysis, grouped Sri Lanka-(Kandy) isolate in the Pacific Indian Oceans (PIO) group. Sri Lanka-(Kandy) isolate of BBTV is classified a new member of PIO group based on analysis of six components of the virus. PMID:27366766

  14. H7 avian influenza virus vaccines protect chickens against challenge with antigenically diverse isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Vaccination has been a critical tool in the control of some avian influenza viruses (AIV) and has been used routinely in Pakistan to help control sporadic outbreaks of highly pathogenic (HP) H7 AIV since 1995. During that time, several AIV isolates were utilized as inactivated vaccines with varying...

  15. Complete Genome Sequence of Human Respiratory Syncytial Virus Isolated in Mexico

    PubMed Central

    Muñoz-Medina, J. E.; Monroy-Muñoz, I. E.; Santos Coy-Arechavaleta, A.; Meza-Chávez, A.; Ángeles-Martínez, J.; Anguiano-Hernández, Y. M.; Santacruz-Tinoco, C. E.; González-Ibarra, J.; Martínez-Miguel, B.; Alvarado-Yaah, J. E.; Palomec-Nava, I. D.; Ortiz-Alcántara, J. M.; Garcés-Ayala, F.; Díaz-Quiñonez, J. A.

    2016-01-01

    Human respiratory syncytial virus (HRSV) is a member of the Paramyxoviridae family, which causes lower respiratory tract infections in neonates and children younger than 5 years. Here, we report the complete genome sequence of HRSV, isolated from a nasopharyngeal swab of a pregnant woman with cardiac complications. PMID:26769933

  16. Complete Genome Sequences of Two Japanese Eel Endothelial Cell-Infecting Virus Strains Isolated in Japan.

    PubMed

    Naoi, Yuki; Okazaki, Sachiko; Katayama, Yukie; Omatsu, Tsutomu; Ono, Shin-Ichi; Mizutani, Tetsuya

    2015-01-01

    Japanese eel endothelial cell-infecting virus (JEECV) causes viral endothelial cell necrosis of eel (VECNE), resulting in severe economic losses in eel aquaculture in Japan. Here, we report the complete genome sequences of two new JEECV strains isolated from farmed Japanese eels. PMID:26564031

  17. Hemagglutinin Receptor Binding of a Human Isolate of Influenza A(H10N8) Virus

    PubMed Central

    Mansour, Mena; Wohlbold, Teddy J.; Ermler, Megan E.; Hirsh, Ariana; Runstadler, Jonathan A.; Fernandez-Sesma, Ana

    2015-01-01

    Three cases of influenza A(H10N8) virus infection in humans have been reported; 2 of these infected persons died. Characterization of the receptor binding pattern of H10 hemagglutinin from avian and human isolates showed that both interact weakly with human-like receptors and maintain strong affinity for avian-like receptors. PMID:26079843

  18. First complete genome sequence of an emerging cucumber green mottle mosaic virus isolate in North America

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The complete genome sequence (6,423 nt) of an emerging Cucumber green mottle mosaic virus (CGMMV) isolate on cucumber in North America was determined through deep sequencing of sRNA and rapid amplification of cDNA ends. It shares 99% nucleotide sequence identity to the Asian genotype, but only 90% t...

  19. Biological and molecular characterization of a US isolate of Hosta virus X

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hosta virus X (HVX) is rapidly becoming a serious pathogen of commercially important hosta plants worldwide. We report here a biological and molecular characterization of a US isolate of HVX, HVX-37. HVX-37 infectivity was tested in 21 hosta cultivars over three growth seasons, and three types of re...

  20. Complete Genome Sequence of Duck Tembusu Virus Isolated from Pekin Ducks in Shanghai, China

    PubMed Central

    Cheng, Yuqiang; Zhang, Chuanpeng; Wang, Hengan; Yan, Yaxian; Ding, Chan

    2015-01-01

    We report here the complete genomic sequence of the duck Tembusu virus (DTMUV) SH001 strain, isolated from Pekin ducks in Shanghai, China, in 2013. The genome of SH001 is 10,990 nucleotides (nt) in length and contains a single open reading frame encoding a putative polyprotein of 3,425 amino acids. PMID:25908131

  1. Complete genome sequence of an Argentinean isolate of Solenopsis invicta virus 3

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genome of an Argentinean isolate of Solenopsis invicta virus 3 (SINV-3ArgSF) obtained from the Santa Fe region of Argentina was sequenced in entirety. Assembly of 9 overlapping fragments yielded a consensus genome sequence 10,386 nucleotides long, excluding the poly(A) tail present on the 3' en...

  2. Molecular analysis of complete genomic sequences of four isolates of Gooseberry vein banding associated virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Presence of Gooseberry vein banding associated virus (GVBaV), a badnavirus in the family Caulimoviridae, is strongly correlated with gooseberry vein banding disease in Ribes spp. In this study, full-length genomic sequences of four GVBaV isolates from different hosts and geographic regions were det...

  3. Complete Genome Sequence of a South Korean Isolate of Habenaria mosaic virus.

    PubMed

    Igori, Davaajargal; Lim, Seungmo; Zhao, Fumei; Baek, Dasom; Moon, Jae Sun

    2016-01-01

    Habenaria mosaic virus (HaMV), a member of the genus Potyvirus in the family Potyviridae, was first discovered from Habenaria radiata in Japan. The complete genomic sequence of a South Korean isolate (PA1) of HaMV infecting Plantago asiatica L. was determined with high-throughput RNA sequencing. PMID:27609926

  4. Complete Genome Sequence of a Variant Pseudorabies Virus Strain Isolated in Central China

    PubMed Central

    Xiang, Shuangshuang; Zhou, Zhi; Hu, Xule; Li, Yingying; Zhang, Chaolin; Wang, Juan; Li, Xiangdong

    2016-01-01

    Pseudorabies virus (PRV) variants have been prevalent in China since 2011 and have caused huge economic losses to the Chinese pig industry. Here, we report the genome sequence of a PRV variant HN1201 that was isolated from diseased animals in central China in 2011. PMID:26988055

  5. Molecular Diversity of Grapevine leafroll-associated virus-2 Isolates in Pacific Northwest Vineyards

    Technology Transfer Automated Retrieval System (TEKTRAN)

    One hundred and five isolates of Grapevine leafroll-associated virus-2 (GLRaV-2) collected from vineyard blocks located in different geographical regions of Washington and Oregon states were characterized based on coat protein (CP) and heat-shock protein 70 homologue (HSP-70h) gene sequences. The re...

  6. Hemagglutinin Receptor Binding of a Human Isolate of Influenza A(H10N8) Virus.

    PubMed

    Ramos, Irene; Mansour, Mena; Wohlbold, Teddy J; Ermler, Megan E; Hirsh, Ariana; Runstadler, Jonathan A; Fernandez-Sesma, Ana; Krammer, Florian

    2015-07-01

    Three cases of influenza A(H10N8) virus infection in humans have been reported; 2 of these infected persons died. Characterization of the receptor binding pattern of H10 hemagglutinin from avian and human isolates showed that both interact weakly with human-like receptors and maintain strong affinity for avian-like receptors. PMID:26079843

  7. Avian influenza virus with Hemagglutinin-Neuraminidase combination H8N8, isolated in Russia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study reports 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....

  8. Replication of parainfluenza (Sendai) virus in isolated rat pulmonary type II alveolar epithelial cells.

    PubMed Central

    Castleman, W. L.; Northrop, P. J.; McAllister, P. K.

    1989-01-01

    The major objectives of this study were to determine whether alveolar type II epithelial cells isolated from rat lung and maintained in tissue culture would support productive replication of parainfluenza type 1 (Sendai) virus and to determine whether isolated type II cells from neonatal (5-day-old) rats that are more susceptible to viral-induced alveolar dysplasia supported viral replication to a greater extent than those from weanling (25-day-old) rats. Isolated and cultured type II cells from neonatal and weanling rats that were inoculated with Sendai virus supported productive replication as indicated by ultrastructural identification of budding virions and viral nucleocapsids in type II cells and by demonstration of rising titers of infectious virus from inoculated type II cell cultures. Alveolar macrophages from neonatal and weanling rats also supported viral replication, although infectious viral titers in macrophage cultures were lower than those from type II cell cultures. Only minor differences were detected between viral titers from neonatal and weanling type II epithelial cell cultures. Higher densities of viral nucleocapsids were observed in neonatal type II cells than in those from weanling rats. The results indicate that isolated type II alveolar epithelial cells support productive replication of parainfluenza virus and that type II cells are probably more efficient in supporting productive viral replication than are alveolar macrophages. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 PMID:2541612

  9. Neurological lesions in chickens experimentally infected with virulent Newcastle disease virus isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Distribution, character, and severity of lesions were evaluated in tissues from the central nervous system of chickens inoculated with 10 different Newcastle disease virus (NDV) isolates: CA 1083, Korea 97-147, Australia (all velogenic viscerotropic); Texas GB and Turkey North Dakota (both velogenic...

  10. SIMILARITIES IN SEED AND APHID TRANSMISSION OF SOYBEAN MOSAIC VIRUS ISOLATES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The strain specificity of transmission of Soybean mosaic virus (SMV) through seed and SMV-induced seed-coat mottling were investigated in field experiments. Six soybean plant introductions (PIs) were inoculated with eight SMV isolates. Transmission of SMV through seed ranged from 0% to 42.6% in see...

  11. First Complete Coding Sequence of a Spanish Isolate of Swine Vesicular Disease Virus

    PubMed Central

    Vázquez-Calvo, Ángela; Saiz, Juan-Carlos; Martín-Acebes, Miguel A.

    2016-01-01

    Swine vesicular disease virus (SVDV) is a porcine pathogen and a member of the Enterovirus genus within the Picornaviridae family. The SVDV genome is composed of a single-stranded RNA molecule of positive polarity. Here, we report the first complete sequence of the coding region of a Spanish SVDV isolate (SPA/1/'93). PMID:26941157

  12. Complete Genome Sequence of a Newcastle Disease Virus Isolated from Wild Peacock (Pavo cristatus) in India

    PubMed Central

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

    2014-01-01

    We report here the complete genome sequence of a Newcastle disease virus (NDV) isolated from a wild peacock. Phylogenetic analysis showed that it belongs to genotype II, class II of NDV strains. This study helps to understand the ecology of NDV strains circulating in a wild avian host of this geographical region during the outbreak of 2012 in northwest India. PMID:24903868

  13. Sequence and virulence comparison of four North American isolates of porcine reproductive and respiratory syndrome virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Considerable genetic, antigenic and virulence differences exist among porcine reproductive and respiratory syndrome virus (PRRSV) isolates and depending on strain, dose and immune status, some farms may be subclinically infected with PRRSV while others experience severe reproductive and/or respirato...

  14. Genome sequences of nine Vesicular Stomatitis Virus isolates from South America

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report nine full-genome sequences of vesicular stomatitis virus obtrained by Illumina next-generation sequencing of RNA, isolated from either cattle epithelial suspensions or cell culture supernatants. Seven of these viral genomes belonged to the New Jersey serotype/species, clade III, while two...

  15. Genetic diversity of Sugarcane bacilliform virus isolates infecting Saccharum spp. in India.

    PubMed

    Karuppaiah, R; Viswanathan, R; Kumar, V Ganesh

    2013-06-01

    Sugarcane bacilliform virus (SCBV), which causes leaf freckle in sugarcane, is a member of the genus Badnavirus. Studies were conducted to characterize SCBV in Saccharum officinarum germplasm and cultivated varieties in India by sequencing the complete genomes of five isolates. Genome lengths ranged from 7,553 to 7,884 nucleotides. Duplications in ORF3 and insertions in the RNase H-domain in some of the isolates were found to contribute to the large size of their genomes. The Indian SCBV isolates share identities of 69-85 % for the complete genomic sequence, indicating wide genetic diversity among them, and share 70-82 % identity with Sugarcane bacilliform Ireng Maleng virus (SCBIMV) and Sugarcane bacilliform Morocco virus (SCBMV), as well as 43-46 % identity with Banana streak virus (BSV) and BSV-related SCBV species from Guadeloupe, indicating that the Indian SCBV isolates are distinct from SCBV isolates reported to date. Irrespective of the region compared, SCBV isolates from India, Australia, and Morocco clustered together. BSV and BSV-related SCBV isolates from Guadeloupe formed another cluster. A phylogenetic analysis based on the partial RT/RNase H-sequence separated SCBV and BSV-related SCBV sequences into 11 SCBV groups viz. SCBV-A to -K. Among the 11 groups, the SCBV sequences separated under H, I, J, and K are newly identified in this study, representing three new species and are tentatively named as SCBBBV, SCBBOV, and SCBBRV. Thus, the PASC and phylogenetic analyses evidenced that the symptoms associated with badnaviruses in sugarcane in India are caused by at least three new species, SCBBBV, SCBBOV, and SCBBRV, besides SCBIMV and SCBMV represented by SCBV-BT and SCBV-Iscam, respectively. PMID:23430710

  16. Receptor specificity in human, avian, and equine H2 and H3 influenza virus isolates.

    PubMed

    Connor, R J; Kawaoka, Y; Webster, R G; Paulson, J C

    1994-11-15

    The receptor specificity of 56 H2 and H3 influenza virus isolates from various animal species has been determined to test the relevance of receptor specificity to the ecology of influenza virus. The results show that the receptor specificity of both H2 and H3 isolates evaluated for sialic acid linkage specificity and inhibition of hemagglutination by horse serum correlates with the species of origin, as postulated earlier for H3 strains based on a limited survey of five human, three avian, and one equine strain. Elucidation of the amino acid sequence of several human H2 receptor variants and analysis of known sequences of H2 and H3 isolates revealed that receptor specificity varies in association with an amino acid change at residues 228 in addition to the change at residue 226 previously documented to affect receptor specificity of H3 but not H1 isolates. Residues 226 and 228 are leucine and serine in human isolates, which preferentially bind sialic acid alpha 2,6-galactose beta 1,4-N-acetyl glucosamine (SA alpha 2,6Gal), and glutamine and glycine in avian and equine isolates, which exhibit specificity for sialic acid alpha-2,3-galactose beta-1,3-N-acetyl galactosamine (SA alpha 2,3Gal). The results demonstrate that the correlation of receptor specificity and species of origin is maintained across both H2 and H3 influenza virus serotypes and provide compelling evidence that influenza virus hosts exert selective pressure to maintain the receptor specificity characteristics of strains isolated from that species. PMID:7975212

  17. Rapid in vitro assessment of the virulence of Newcastle disease virus isolates using the ligase chain reaction.

    PubMed

    Collins, M S; Govey, S J; Alexander, D J

    2003-09-01

    The ligase chain reaction was used to assess the virulence of isolates of Newcastle disease virus. In the main study, 18/18 virulent isolates whose nucleotide sequences that code for the cleavage site and fusor peptide regions were known, successfully ligated oligonucleotides in a primer mix for virulent viruses termed VPM. Five of these isolates yielded a more intense ligated product with a second primer mix for virulent viruses called VPM1. No ligation was evident with eight avirulent isolates in tests with VPM or VPM1, however, each of these viruses did yield a strong ligated product with the primer mix for avirulent viruses (AVPM) as did one virulent isolate considered to be a mixture. Two virulent Australian isolates, 1238/1998 and 1248/1998, showed low but seemingly specific ligation with AVPM. In a blind study, 8/9 virulent isolates whose sequences were unknown ligated primers in VPM. Three avirulent and one virulent isolate, the latter again probably a mixture, ligated primers in AVPM. Ligation of oligonucleotides in VPM and AVPM was detectable in mixtures where virulent and avirulent isolates represented 0.1% and 0.01% by volume respectively of the viral population. The results indicate that LCR offers a potential in vitro alternative to current in vivo tests for virulence determination of Newcastle disease virus isolates. PMID:14505095

  18. A noda-like virus isolated from the sweetpotato pest spodoptera eridania (Cramer) (Lep.; noctuidae)

    PubMed

    Zeddam; Rodriguez; Ravallec; Lagnaoui

    1999-11-01

    A small isometric virus has been isolated from larvae of the sweetpotato pest Spodoptera eridania (Cramer) collected near Pariacoto, Ancash province, Peru. It is designated the Pariacoto virus (PaV). In addition to its high pathogenicity on its natural host Spodoptera eridania, PaV was found to replicate in Spodoptera ochrea (Hampson) larvae but not in Spodoptera frugiperda (Smith) larvae. The size of the viral particle was estimated to be about 30 nm in diameter. Polyacrylamide gel electrophoresis showed a protein of approximately 40.5 kDa. After agarose gel electrophoresis, the viral genome appeared to be bipartite RNA. Gel immunodiffusion tests showed no serological relationship between PaV and Nodamura virus, the type species for insect nodaviruses. Electron microscopy confirmed that viral replication occurs in the cytoplasm. These properties are similar to those of other members of family Nodaviridae, to which the virus is currently assigned. Copyright 1999 Academic Press. PMID:10534414

  19. Wolbachia Blocks Currently Circulating Zika Virus Isolates in Brazilian Aedes aegypti Mosquitoes.

    PubMed

    Dutra, Heverton Leandro Carneiro; Rocha, Marcele Neves; Dias, Fernando Braga Stehling; Mansur, Simone Brutman; Caragata, Eric Pearce; Moreira, Luciano Andrade

    2016-06-01

    The recent association of Zika virus with cases of microcephaly has sparked a global health crisis and highlighted the need for mechanisms to combat the Zika vector, Aedes aegypti mosquitoes. Wolbachia pipientis, a bacterial endosymbiont of insect, has recently garnered attention as a mechanism for arbovirus control. Here we report that Aedes aegypti harboring Wolbachia are highly resistant to infection with two currently circulating Zika virus isolates from the recent Brazilian epidemic. Wolbachia-harboring mosquitoes displayed lower viral prevalence and intensity and decreased disseminated infection and, critically, did not carry infectious virus in the saliva, suggesting that viral transmission was blocked. Our data indicate that the use of Wolbachia-harboring mosquitoes could represent an effective mechanism to reduce Zika virus transmission and should be included as part of Zika control strategies. PMID:27156023

  20. Novel forms of woodchuck hepatitis virus DNA isolated from chronically infected woodchuck liver nuclei.

    PubMed Central

    Rogler, C E; Summers, J

    1982-01-01

    We cloned several unique forms of woodchuck hepatitis virus, a DNA virus closely related to hepatitis B virus, from a chronically infected woodchuck liver. Each of the three clones contained more than two genome equivalents of viral sequences with extensive rearrangements and no detectable cellular sequences. From the frequency by which they were isolated from a library of recombinant clones, we estimate that they are present in approximately one copy per cell. Of a total of 11 sites at which rearrangements were mapped in the clones, 10 occurred between segments of opposite polarity, and 1 occurred between segments of the same polarity. The possible significance of these findings to the persistence of virus production in infected cells is discussed. Images PMID:6294334

  1. Comparison of the pathogenicity of Nipah virus isolates from Bangladesh and Malaysia in the Syrian hamster.

    PubMed

    DeBuysscher, Blair L; de Wit, Emmie; Munster, Vincent J; Scott, Dana; Feldmann, Heinz; Prescott, Joseph

    2013-01-01

    Nipah virus is a zoonotic pathogen that causes severe disease in humans. The mechanisms of pathogenesis are not well described. The first Nipah virus outbreak occurred in Malaysia, where human disease had a strong neurological component. Subsequent outbreaks have occurred in Bangladesh and India and transmission and disease processes in these outbreaks appear to be different from those of the Malaysian outbreak. Until this point, virtually all Nipah virus studies in vitro and in vivo, including vaccine and pathogenesis studies, have utilized a virus isolate from the original Malaysian outbreak (NiV-M). To investigate potential differences between NiV-M and a Nipah virus isolate from Bangladesh (NiV-B), we compared NiV-M and NiV-B infection in vitro and in vivo. In hamster kidney cells, NiV-M-infection resulted in extensive syncytia formation and cytopathic effects, whereas NiV-B-infection resulted in little to no morphological changes. In vivo, NiV-M-infected Syrian hamsters had accelerated virus replication, pathology and death when compared to NiV-B-infected animals. NiV-M infection also resulted in the activation of host immune response genes at an earlier time point. Pathogenicity was not only a result of direct effects of virus replication, but likely also had an immunopathogenic component. The differences observed between NiV-M and NiV-B pathogeneis in hamsters may relate to differences observed in human cases. Characterization of the hamster model for NiV-B infection allows for further research of the strain of Nipah virus responsible for the more recent outbreaks in humans. This model can be used to study NiV-B pathogenesis, transmission, and countermeasures that could be used to control outbreaks. PMID:23342177

  2. Isolation and Characterization of a Single-Stranded DNA Virus Infecting Chaetoceros lorenzianus Grunow▿

    PubMed Central

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

    2011-01-01

    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

  3. Comparison of the Pathogenicity of Nipah Virus Isolates from Bangladesh and Malaysia in the Syrian Hamster

    PubMed Central

    DeBuysscher, Blair L.; de Wit, Emmie; Munster, Vincent J.; Scott, Dana; Feldmann, Heinz; Prescott, Joseph

    2013-01-01

    Nipah virus is a zoonotic pathogen that causes severe disease in humans. The mechanisms of pathogenesis are not well described. The first Nipah virus outbreak occurred in Malaysia, where human disease had a strong neurological component. Subsequent outbreaks have occurred in Bangladesh and India and transmission and disease processes in these outbreaks appear to be different from those of the Malaysian outbreak. Until this point, virtually all Nipah virus studies in vitro and in vivo, including vaccine and pathogenesis studies, have utilized a virus isolate from the original Malaysian outbreak (NiV-M). To investigate potential differences between NiV-M and a Nipah virus isolate from Bangladesh (NiV-B), we compared NiV-M and NiV-B infection in vitro and in vivo. In hamster kidney cells, NiV-M-infection resulted in extensive syncytia formation and cytopathic effects, whereas NiV-B-infection resulted in little to no morphological changes. In vivo, NiV-M-infected Syrian hamsters had accelerated virus replication, pathology and death when compared to NiV-B-infected animals. NiV-M infection also resulted in the activation of host immune response genes at an earlier time point. Pathogenicity was not only a result of direct effects of virus replication, but likely also had an immunopathogenic component. The differences observed between NiV-M and NiV-B pathogeneis in hamsters may relate to differences observed in human cases. Characterization of the hamster model for NiV-B infection allows for further research of the strain of Nipah virus responsible for the more recent outbreaks in humans. This model can be used to study NiV-B pathogenesis, transmission, and countermeasures that could be used to control outbreaks. PMID:23342177

  4. Population structure of blueberry mosaic associated virus: Evidence of reassortment in geographically distinct isolates.

    PubMed

    Thekke-Veetil, Thanuja; Polashock, James J; Marn, Mojca V; Plesko, Irena M; Schilder, Annemiek C; Keller, Karen E; Martin, Robert R; Tzanetakis, Ioannis E

    2015-04-01

    The population structure of blueberry mosaic associated virus (BlMaV), a putative member of the family Ophioviridae, was examined using 61 isolates collected from North America and Slovenia. The studied isolates displayed low diversity in the movement and nucleocapsid proteins and low ratios of non-synonymous to synonymous nucleotide substitutions, indicative of strong purifying selection. Phylogenetic analyses revealed grouping primarily based on geography with some isolates deviating from this rule. Phylogenetic incongruence in the two regions, coupled with detection of reassortment events, indicated the possible role of genetic exchange in the evolution of BlMaV. PMID:25733053

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

    PubMed Central

    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

    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

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

    PubMed

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

    2014-12-01

    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

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

    PubMed

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

    2002-01-01

    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

  8. Genotypic and pathotypic characterization of Newcastle disease virus isolated from racing pigeons in China.

    PubMed

    Liu, Mengda; Qu, Yajin; Wang, Fangkun; Liu, Sidang; Sun, Honglei

    2015-07-01

    A Newcastle disease virus (NDV) isolated from an outbreak in racing pigeons in China was characterized in this study. Complete gene of the NDV isolate was sequenced and phylogenetic analysis. Pathogenicity experiment was carried out in pigeons, chickens, and ducks. Phylogenetic analysis revealed that the strain clustered with the Class II viruses, has highly phylogenetically similar to NDV strains isolated from pigeons in China, but was distant from the viruses prevalence in chickens and vaccine strains used in China. The deduced amino acid sequence of the cleavage site of the fusion (F) protein confirmed that the isolate contained the virulent motif (112)RRQKRF(117) at the cleavage site, but it caused no appearance disease in chickens and ducks. However, the isolate had virulence in pigeons, resulting in severe nervous signs and highly mortality. Pigeons were considered as a potential source of NDV infection and disease for commercial poultry flocks. Therefore, new vaccines to prevent the NDV infection in the pigeon flocks should be developed as soon as possible, and strict biosecurity measures should be taken to reduce the risk of pigeon Newcastle disease outbreaks. PMID:25877412

  9. Isolation, molecular and phylogenetic analysis of encephalomyocarditis virus strain GS01 in China.

    PubMed

    Feng, Ruofei; Zhang, Haixia; Wei, Jia; Li, Xiangrong; Xie, Jingying; Li, Mingsheng; Qiao, Zilin; Feng, Yuping; Ma, Zhongren

    2015-03-01

    Encephalomyocarditis virus (EMCV) is a small non-enveloped, single-stranded RNA virus. It can infect many host species and cause acute myocarditis and respiratory failure in piglets, reproductive failure in pregnant sows. Diseases caused by EMCV currently affect the swine industry worldwide. In this study, an EMCV strain was isolated from an aborted fetus in western China. It was identified by reverse-transcription polymerase chain reaction (RT-PCR) and genome sequencing. The subsequent results showed that the virus could produce a specific cytopathic effect on BHK-21 cells and could cause severe clinical symptoms and pathological changes in mice. Complete genome sequencing and multiple sequence alignment indicated that the GS01 strain was 79.9-99.9% identical with other isolates worldwide. Phylogenetic analysis showed that EMCV isolates fell into five clusters: lineage 1, 2, 3, 4, and 5 based on the nucleotide sequences of the entire ORF and VP3/VP1 junction, as well as 3D gene. GS01 isolate was grouped into lineage 1. The results of this study confirmed that an EMCV strain GS01 isolated from an aborted pig fetus in western China was fatal to mice and provided new epidemiologic data on EMCV in China. PMID:25497352

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

    PubMed

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

    2014-06-01

    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

  11. Complete nucleotide sequence of a Spanish isolate of alfalfa mosaic virus: evidence for additional genetic variability.

    PubMed

    Parrella, Giuseppe; Acanfora, Nadia; Orílio, Anelise F; Navas-Castillo, Jesús

    2011-06-01

    Alfalfa mosaic virus (AMV) is a plant virus that is distributed worldwide and can induce necrosis and/or yellow mosaic on a large variety of plant species, including commercially important crops. It is the only virus of the genus Alfamovirus in the family Bromoviridae. AMV isolates can be clustered into two genetic groups that correlate with their geographic origin. Here, we report for the first time the complete nucleotide sequence of a Spanish isolate of AMV found infecting Cape honeysuckle (Tecoma capensis) and named Tec-1. The tripartite genome of Tec-1 is composed of 3643 nucleotides (nt) for RNA1, 2594 nt for RNA2 and 2037 nt for RNA3. Comparative sequence analysis of the coat protein gene revealed that the isolate Tec-1 is distantly related to subgroup I of AMV and more closely related to subgroup II, although forming a distinct phylogenetic clade. Therefore, we propose to split subgroup II of AMV into two subgroups, namely IIA, comprising isolates previously included in subgroup II, and IIB, including the novel Spanish isolate Tec-1. PMID:21327783

  12. Phylogenetic analysis of Chinese sheeppox and goatpox virus isolates

    PubMed Central

    2012-01-01

    Background Sheeppox virus (SPPV) and goatpox virus (GTPV), members of the Capripoxvirus genus of the Poxviridae family are causative agents of sheep pox and goat pox respectively, which are important contagious diseases and endemic in central and northern Africa, the Middle and Far East, and the Indian sub-continent. Both sheep pox and goat pox can cause wool and hide damage, and reduce the production of mutton and milk, which may result in significant economic losses and threaten the stockbreeding. In this study, three SPPVs and two GTPVs were collected from China in 2009 and 2011. We described the sequence features and phylogenetic analysis of the P32 gene, GPCR gene and RPO30 gene of the SPPVs and GTPVs to reveal their genetic relatedness. Results Sequence and phylogenetic analysis showed that there was a close relationship among SPPV/GanS/2/2011/China, SPPV/GanS/1/2011/China and SPPV/NingX/2009/China. They were clustered on the same SPPV clade. GTPV/HuB/2009/China and GS-V1 belonged to the GTPV lineage. GS-V1 was closely related to other GTPV vaccine strains. GTPV/HuB/2009/China and GS-V1 were clustered with GTPVs from China and some southern Asian countries. Conclusion This study may expand the datum for spread trend research of Chinese SPPVs and GTPVs, meanwhile provide theoretical references to improve the preventive and control strategy. PMID:22264255

  13. [Characteristics of virus double-stranded RNA, isolated from microscopic fungi parasitizing on sugar beet].

    PubMed

    Mel'nychuk, M D; Spyrydonov, V H; Oleksiienko, I P

    2005-01-01

    We have carried out comparative studies of double-stranded RNA (dsRNA) of viral nature isolated from sugar beet leaves and from mycelium of microscopic fungi using different methods such as PAAG electrophoresis and by polymerase chain reaction (PCR). It was shown that the fragments of dsRNA from sugar beet leaves and from mycelium microscopic fungi had the identical electrophoretic pattern and the same size (1.8 and 2.0 kbp). Using PCR technique it was shown, that isolated dsRNA have a common template for amplification. Electron microscopy of PCR-positive mycelium allows us to detect the virus particles of the spherical form with diameter 30-40 nm. The obtained data confirm our previous suppositions, concerning the belonging of isolated dsRNAs (size 1.8 and 2.0 kbp) to new mycovirus targeted a microscopic fungus, instead of beet cryptic viruses. PMID:16250236

  14. Isolation of novel variants of infectious bursal disease virus from different outbreaks in Northeast India.

    PubMed

    Morla, Sudhir; Deka, Pankaj; Kumar, Sachin

    2016-04-01

    Infectious bursal disease virus (IBDV) is a highly infectious disease of young chicken that predominantly affects the immune system. In the present study, we are reporting first comprehensive study of IBDV outbreaks from the Northeastern part of India. Northeast India shares a porous border with four different countries; and as a rule any outbreak in the neighboring countries substantially affects the poultry population in the adjoining states. Nucleotide sequence analysis of the VP2 gene of the IBDV isolates from the Northeastern part of India suggested the extreme virulent nature of the virus. The virulent marker amino acids (A222, I242, Q253, I256 and S299) in the hypervariable region of the Northeastern isolates were found identical with the reported very virulent strains of IBDV. A unique insertion of I/L294V was recorded in all the isolates of the Northeastern India. The study will be useful in understanding the circulating pathotypes of IBDV in India. PMID:26854869

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

    PubMed

    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

    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

  16. Molecular characterization of Dasheen mosaic virus isolates infecting edible aroids in India.

    PubMed

    Babu, B; Hegde, V

    2014-01-01

    Dasheen mosaic virus (DsMV) infecting three major edible aroids namely Amorphophallus paeoniifolius, Colocasia esculenta, and Xanthosoma sagittifolium cultivated in India was characterized. Infected plants showing typical DsMV symptoms were subjected to reverse transcription-polymerase chain reaction, and an amplification of a 963 bp fragment which encoded the coat protein (CP) gene was obtained. BLAST analysis of the cloned DNA amplicon revealed the identity of the virus to be that of DsMV. Sequence identity matrix of the nucleotide sequences among the three isolates showed that the DsMV isolate infecting A. paeoniifolius and C. esculenta shared an identity as high as 93%, while the DsMV isolate from X. sagittifolium shared an identity of only 73% and 76% with the DsMV isolates from A. paeoniifolius and C. esculenta, respectively. Comparative analysis of the coat protein of the three DsMV isolates showed the presence of DVG motif (A. paeoniifolius and C. esculenta) and DTG motif in X. sagittifolium and several varying potential threonine and asparagine rich N-glycosylation motifs. Single amino acid substitution of the several conserved motifs occurs in all the three DsMV isolates. This is the first characterization of DsMV isolates infecting A. paeoniifolius, C. esculenta, and X. sagittifolium plants in India. PMID:24717027

  17. Phylogenetic analysis of bluetongue virus serotype 4 field isolates from Argentina.

    PubMed

    Legisa, D; Gonzalez, F; De Stefano, G; Pereda, A; Dus Santos, M J

    2013-03-01

    Bluetongue is an insect-transmitted viral disease of ruminant species, which represents a major barrier to the international trade of animals and their products. Bluetongue virus (BTV) has a genome composed of ten linear segments of dsRNA, which code for at least ten different viral proteins. In South America, serological evidence for the presence of BTV has been found in Peru, Argentina, Brazil, Ecuador and Chile. Brazil and Argentina are the only South American countries where BTV has been isolated. In Brazil, only one BTV isolate, serotype 12, has been reported, whereas in Argentina five BTV serotype 4 isolates have been obtained from cattle without clinical signs. Three of these five isolates were isolated during 1999-2001, whereas two of them were obtained as part of the present work. This study describes sequence comparisons and phylogenetic analyses of segment (Seg)-2, Seg-3, Seg-6, Seg-7 and Seg-10 of the first Argentinian field isolates of BTV. The analysis of Seg-2 and Seg-6 resulted in a single cluster of Argentinian sequences into the serotype 4 clade. In addition, the Argentinian sequences grouped within the nucleotype A clade, along with reference strains. The analysis of Seg-3, Seg-7 and Seg-10 showed that the Argentinian isolates grouped into the western topotype, indicating that the circulating virus had an African/European origin. Phylogenetic analysis revealed that the Argentinian sequences present a South American genetic identity, suggesting an independent lineage evolution. PMID:23152367

  18. Biochemical and antigenic properties of the first isolates of infectious hematopoietic necrosis virus from salmonid fish in Europe

    USGS Publications Warehouse

    Arkush, K.D.; Bovo, G.; deKinkelin, P.; Winton, J.R.; Wingfield, W.H.; Hedrick, R.P.

    1989-01-01

    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 SDS-PAGE. An analysis of the antigenic relatedness of the European isolates to representative strains from North America showed that they were clearly different from viruses obtained from salmonids in California. The RB/B5 MAb, which was developed against virus isolated from adult steelhead (anadromous rainbow trout) reared in central Oregon, neutralized all isolates examined. The 193–110/B4 MAb, developed against IHNV isolated from infected yearling rainbow trout in southern Idaho, neutralized all isolates tested except those from California. The SRCV/A4 MAb, developed against Sacramento River chinook virus (SRCV) isolated from adult spring chinook salmon O. tshawytscha in central California, was the least reactive, and strong neutralization was observed only with the SRCV strain of IHNV from California. However, partial reactivity of the virus isolates from France with the SRCV/A4 MAb distinguished them from the virus recovered from salmonids in Italy.

  19. Clustering of classical swine fever virus isolates by codon pair bias

    PubMed Central

    2011-01-01

    Background The genetic code consists of non-random usage of synonymous codons for the same amino acids, termed codon bias or codon usage. Codon juxtaposition is also non-random, referred to as codon context bias or codon pair bias. The codon and codon pair bias vary among different organisms, as well as with viruses. Reasons for these differences are not completely understood. For classical swine fever virus (CSFV), it was suggested that the synonymous codon usage does not significantly influence virulence, but the relationship between variations in codon pair usage and CSFV virulence is unknown. Virulence can be related to the fitness of a virus: Differences in codon pair usage influence genome translation efficiency, which may in turn relate to the fitness of a virus. Accordingly, the potential of the codon pair bias for clustering CSFV isolates into classes of different virulence was investigated. Results The complete genomic sequences encoding the viral polyprotein of 52 different CSFV isolates were analyzed. This included 49 sequences from the GenBank database (NCBI) and three newly sequenced genomes. The codon usage did not differ among isolates of different virulence or genotype. In contrast, a clustering of isolates based on their codon pair bias was observed, clearly discriminating highly virulent isolates and vaccine strains on one side from moderately virulent strains on the other side. However, phylogenetic trees based on the codon pair bias and on the primary nucleotide sequence resulted in a very similar genotype distribution. Conclusion Clustering of CSFV genomes based on their codon pair bias correlate with the genotype rather than with the virulence of the isolates. PMID:22126254

  20. Isolation and identification of a bovine viral diarrhea virus from sika deer in china

    PubMed Central

    2011-01-01

    Background Bovine viral diarrhea virus (BVDV) infections continue to cause significantly losses in the deer population. Better isolation and identification of BVDV from sika deer may contribute significantly to the development of prophylactic therapeutic, and diagnostic reagents as well as help in prevention and control of BVDV. However, isolation and identification of BVDV from sika deer is seldom reported in literature. In this study, we collected some samples according to clinical sign of BVDV to isolation and identification of BVDV from sika deer. Results we isolated a suspected BVDV strain from livers of an aborted fetus from sika deer in Changchun (China) using MDBK cell lines, named as CCSYD strain, and identified it by cytopathic effect (CPE), indirect immunoperoxidase test (IPX) and electron microscopy(EM). The results indicated that this virus was BVDV by a series of identification. The structural proteins E0 gene was cloned and sequenced. The obtained E0 gene sequence has been submitted to GenBank with the accession number: FJ555203. Alignment with other 9 strains of BVDV, 7 strains of classical swine fever virus (CSFV) and 3 strains of border disease virus(BDV) in the world, showed that the homology were 98.6%-84.8%, 76.0%-74.7%, 76.6%-77.0% for nucleotide sequence, respectively. The phylogenetic analysis indicated that new isolation and identification CCSYD strain belonged to BVDV1b. Conclusion To the best of our knowledge, this is the first report that BVDV was isolated and identified in sika deer. This current research contributes development new BVDV vaccine to prevent and control of BVD in sika deer. PMID:21352530

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

    PubMed

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

    2012-10-01

    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

  2. Molecular Characterization of Geographically Different Banana bunchy top virus Isolates in India.

    PubMed

    Selvarajan, R; Mary Sheeba, M; Balasubramanian, V; Rajmohan, R; Dhevi, N Lakshmi; Sasireka, T

    2010-10-01

    Banana bunchy top disease (BBTD) caused by Banana bunchy top virus (BBTV) is one of the most devastating diseases of banana and poses a serious threat for cultivars like Hill Banana (Syn: Virupakshi) and Grand Naine in India. In this study, we have cloned and sequenced the complete genome comprised of six DNA components of BBTV infecting Hill Banana grown in lower Pulney hills, Tamil Nadu State, India. The complete genome sequence of this hill banana isolate showed high degree of similarity with the corresponding sequences of BBTV isolates originating from Lucknow, Uttar Pradesh State, India, and from Fiji, Egypt, Pakistan, and Australia. In addition, sixteen coat protein (CP) and thirteen replicase genes (Rep) sequences of BBTV isolates collected from different banana growing states of India were cloned and sequenced. The replicase sequences of 13 isolates showed high degree of similarity with that of South Pacific group of BBTV isolates. However, the CP gene of BBTV isolates from Shervroy and Kodaikanal hills of Tamil Nadu showed higher amino acid sequence variability compared to other isolates. Another hill banana isolate from Meghalaya state had 23 nucleotide substitutions in the CP gene but the amino acid sequence was conserved. This is the first report of the characterization of a complete genome of BBTV occurring in the high altitudes of India. Our study revealed that the Indian BBTV isolates with distinct geographical origins belongs to the South Pacific group, except Shervroy and Kodaikanal hill isolates which neither belong to the South Pacific nor the Asian group. PMID:23637489

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

    PubMed Central

    2014-01-01

    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

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

    PubMed Central

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

    2011-01-01

    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

  5. Phylogenetic and Pathogenic Analyses of Avian Influenza A H5N1 Viruses Isolated from Poultry in Vietnam

    PubMed Central

    Li, Yanbing; Jiang, Yongping; Liu, Liling; Chen, Hualan

    2012-01-01

    Despite great efforts to control the infection of poultry with H5N1 viruses, these pathogens continue to evolve and spread in nature, threatening public health. Elucidating the characteristics of H5N1 avian influenza virus will benefit disease control and pandemic preparation. Here, we sequenced the genomes of 15 H5N1 avian influenza viruses isolated in Vietnam in 2006 and 2007 and performed phylogenetic analyses to compare these sequences with those of other viruses available in the public databases. Molecular characterization of the H5N1 viruses revealed that seven genetically distinct clades of H5N1 viruses have appeared in Vietnam. Clade 2.3.4 viruses existed in Vietnam as early as 2005. Fifteen viruses isolated during 2006 and 2007 belonged to clade 1 and clade 2.3.4, and were divided into five genotypes. Reassortants between the clade 1 and clade 2.3.4 viruses were detected in both North and South Vietnam. We also assessed the replication and pathogenicity of these viruses in mice and found that these isolates replicated efficiently and exhibited distinct virulence in mice. Our results provide important information regarding the diversity of H5N1 viruses in nature. PMID:23226433

  6. Virus isolation from saliva and salivary glands of cattle naturally infected with paralytic rabies.

    PubMed

    Delpietro, H A; Larghi, O P; Russo, R G

    2001-02-16

    The infectivity of saliva, salivary and mammary glands, muscle, lung, kidney and liver of 87 cattle infected with paralytic rabies (positive viral isolation from brains) was studied. Fifty percent dilutions of saliva and tissue samples were inoculated intracerebrally into 10- to 15-day-old mice. Viral isolation in mice was confirmed by direct rabies fluorescent-antibody test and the antigenic variant of the isolates characterized by monoclonal antibodies. Rabies virus was isolated from 4.6% of salivary glands and from 1.6% of saliva samples. The rest of the peripheral tissues were negative. Cerebral and peripheral isolates belonged to vampire-bat antigenic variants. These results indicate that cattle infected by vampire bats may be a source of infection for man. The infection risk would depend on the type of contact between rabid cattle and man. PMID:11182465

  7. Identification of a bovine viral diarrhea virus 2 isolated from cattle in China.

    PubMed

    Zhu, L Q; Ren, M; Lin, Y Q; Ding, X Y; Zhang, G P; Zhao, X; Zhu, G Q

    2009-01-01

    The identification and characterization of bovine viral diarrhea virus 2 (BVDV-2) strain SD-06 isolated from cattle in China is reported. We performed sequence analysis of 5'-untranslated region (5'-UTR) and E2 sequences and the identity at the nucleotide and amino acid level indicated that the isolate was closely related to BVDV-2. The BVDV-2 strain New York'93 showed the highest sequence homology with the isolate SD-06. The phylogenetic analysis revealed that the isolate SD-06 belonged to BVDV-2a subtype. Furthermore, immunofluorescence assay with the monoclonal antibody specific for BVDV-2 glycoprotein E2 confirmed this identification. Thus, the strain SD-06 was the first isolate of BVDV-2 identified in China. PMID:19537915

  8. A new subgenotype 2.1d isolates of classical swine fever virus in China, 2014.

    PubMed

    Zhang, Hongliang; Leng, Chaoliang; Feng, Liping; Zhai, Hongyue; Chen, Jiazeng; Liu, Chunxiao; Bai, Yun; Ye, Chao; Peng, Jinmei; An, Tongqing; Kan, Yunchao; Cai, Xuehui; Tian, Zhijun; Tong, Guangzhi

    2015-08-01

    The lapinized attenuated vaccine against classical swine fever (CSF) has been used in China for over half a century and has generally prevented large-scale outbreaks in recent years. However, since late 2014, a large number of new cases of CSF were detected in many immunized pig farms in China. Several of these CSV viruses were isolated and characterized. Phylogenetic and genomic sequence analyses indicate that these new isolates, as well as some reference isolates, form a new subgenotype named 2.1d, and share several consistent molecular characteristics. Since these new isolates emerged in disparate geographic regions within 5 months, this suggests that these isolates may be widespread. Given that current vaccines do not appear to provide effective protection against this new subgenotype, further investigation of these strains is urgently needed. PMID:26031602

  9. [The Isolation and Identification of Infectious Bronchitis Virus PTFY Strain in Muscovy Ducks].

    PubMed

    Wu, Xiaoping; Pan, Shulei; Zhou, Wuduo; Wu, Yijiang; Huang, Yifan; Wu, Baocheng

    2016-03-01

    In July 2009, some farms of breeding Muscovy ducks on the peak of egg laying suffered the decrease of hatching rate and the quality of the eggs showing low mortality and no evident respiratory symptoms. The swelling and congestive ovary was visible after autopsy. This study was brought out for the diagnosis of these cases. The virus was isolated and identified by the methods of virus culture in chicken embryo, physical and chemical properties test, hemagglutinin test, NDV (Newcastle diseases Virus) interference test, electron microscope observation, pathogenicity test and the gene sequence analysis. The results indicated the virus showed the characters of inducing dwarf embryo after inocubation, the sensibility to lipid solvent and the hemagglutination capacity after pancreatic enzyme treatment, the typical morphology of coronavirus, the interference to NDV replication and the homology among 84.7% - 99% of the particial N gene sequences to the reference IBV (Avian infectious bronchitis virus) strains. The strain was identified as IBV isolate and this study confirmed the pathogenicity of IBV to Muscovy ducks. PMID:27396165

  10. Molecular analysis of the complete genomic sequences of four isolates of Gooseberry vein banding associated virus.

    PubMed

    Xu, Donglin; Mock, Ray; Kinard, Gary; Li, Ruhui

    2011-08-01

    The presence of Gooseberry vein banding associated virus (GVBaV), a badnavirus in the family Caulimoviridae, is strongly correlated with gooseberry vein banding disease in Ribes spp. In this study, full-length genomic sequences of four GVBaV isolates from different hosts and geographic regions were determined to be 7649-7663 nucleotides. These isolates share identities of 96.4-97.3% for the complete genomic sequence, indicating low genetic diversity among them. The GVBaV genome contains three open reading frames (ORFs) on the plus strand that potentially encode proteins of 26, 16, and 216 kDa. The size and organization of GVBaV ORFs 1-3 are similar to those of most other badnaviruses. The putative amino acid sequence of GVBaV ORF 3 contained motifs that are conserved among badnavirus proteins including aspartic protease, reverse transcriptase, and ribonuclease H. The highly conserved putative plant tRNA(met)-binding site is also present in the 935-bp intergenic region of GVBaV. The identities of the genomic sequences of GVBaV and other badnaviruses range from 49.1% (Sugarcane bacilliform Mor virus) to 51.7% (Pelargonium vein banding virus, PVBV). Phylogenetic analysis using the amino acid sequence of the ORF 3 putative protein shows that GVBaV groups most closely to Dioscorea bacilliform virus, PVBV, and Taro bacilliform virus. These results confirm that GVBaV is a pararetrovirus of the genus Badnavirus in the family Caulimoviridae. PMID:21533750

  11. Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys

    PubMed Central

    2010-01-01

    Background The highly pathogenic H5N1 is a major avian pathogen that crosses species barriers and seriously affects humans as well as some mammals. It mutates in an intensified manner and is considered a potential candidate for the possible next pandemic with all the catastrophic consequences. Methods Nasal swabs were collected from donkeys suffered from respiratory distress. The virus was isolated from the pooled nasal swabs in specific pathogen free embryonated chicken eggs (SPF-ECE). Reverse transcriptase polymerase chain reaction (RT-PCR) and sequencing of both haemagglutingin and neuraminidase were performed. H5 seroconversion was screened using haemagglutination inhibition (HI) assay on 105 donkey serum samples. Results We demonstrated that H5N1 jumped from poultry to another mammalian host; donkeys. Phylogenetic analysis showed that the virus clustered within the lineage of H5N1 from Egypt, closely related to 2009 isolates. It harboured few genetic changes compared to the closely related viruses from avian and humans. The neuraminidase lacks oseltamivir resistant mutations. Interestingly, HI screening for antibodies to H5 haemagglutinins in donkeys revealed high exposure rate. Conclusions These findings extend the host range of the H5N1 influenza virus, possess implications for influenza virus epidemiology and highlight the need for the systematic surveillance of H5N1 in animals in the vicinity of backyard poultry units especially in endemic areas. PMID:20398268

  12. Isolation of avian pneumovirus from mallard ducks that is genetically similar to viruses isolated from neighboring commercial turkeys.

    PubMed

    Shin, Hyun-Jin; Nagaraja, Kakambi V; McComb, Brian; Halvorson, David A; Jirjis, Faris F; Shaw, Daniel P; Seal, Bruce S; Njenga, M Kariuki

    2002-02-26

    Our earlier studies demonstrating avian pneumovirus (APV) RNA in wild geese, sparrows, swallows, starlings and mallard ducks suggested that wild birds might be involved in the circulation of APV in the United States. To determine whether turkey virus can be transmitted to the free flying birds, we placed APV-negative mallard ducks next to a turkey farm experiencing a severe APV outbreak and in an area with a large population of waterfowls. The sentinel ducks did not develop clinical APV disease but infectious APV (APV/MN-12) was recovered from choanal swabs after 2 weeks, and anti-APV antibodies detected after 4 weeks. Four APV isolates recovered from the neighboring turkeys that were experiencing an APV outbreak at the same time shared 95-99% nucleotide identity and 97-99% predicted amino acid identity with the duck isolate. In addition experimental infection of turkey poults with APV/MN-12 resulted in detection of viral RNA in nasal turbinates and APV-specific IgG in serum. These results indicate that the APV isolates from turkeys and ducks shared a common source, and the viruses from different avian species can cross-infect. PMID:11864753

  13. Isolation and full-genome sequence of two reticuloendotheliosis virus strains from mixed infections with Marek's disease virus in China.

    PubMed

    Bao, Ke-yan; Zhang, Yan-ping; Zheng, Hui-wen; Lv, Hong-chao; Gao, Yu-long; Wang, Jing-fei; Gao, Hong-lei; Qi, Xiao-le; Cui, Hong-yu; Wang, Yong-qiang; Ren, Xian-gang; Wang, Xiao-mei; Liu, Chang-jun

    2015-06-01

    Reticuloendotheliosis virus (REV), classified as a gammaretrovirus, has a variety of hosts, including chickens, ducks, geese, turkeys, and wild birds. REV causes a series of pathological syndromes, especially the immunosuppression of the host, which may lead to an increased susceptibility to other pathogens, thus greatly damaging the poultry industry. Mixed infections of REV and Marek's disease virus (MDV) have been reported in many countries, including China. Previous reports revealed that MDV vaccines were not efficacious, and even less-virulent MDV strains would cause some losses due to mixed infections with REV. Additionally, contaminants in the MDV vaccine might be the main source of REV. In this study, two clinical samples were collected from two flocks of chickens that were diagnosed with MDV. Subsequently, two REV isolates were obtained from the clinical samples. The isolates, named CY1111 and SY1209, were further confirmed through an indirect immunofluorescence assay and electron microscopy. Complete genome sequences of the two REV strains were determined to test the relationship between them and other REV strains. Phylogenetic trees showed that the two REV strains were closely related to most REV strains that were isolated from a variety of hosts. Therefore, REVs might spread freely among these hosts under natural conditions. Additionally, most REV strains in China were in the same clade. The present work offers some information regarding REV in China. PMID:25850423

  14. Characterization of a novel H3N2 influenza virus isolated from domestic ducks in China.

    PubMed

    Li, Chong; Yu, Meng; Liu, Litao; Sun, Honglei

    2016-08-01

    Cases of human infection with a novel H7N9 avian influenza virus (AIV) were first reported in March 2013, which caused 115 deaths within a single year. Beyond that, other subtypes of H7 AIV were isolated from poultry in eastern China during the same period, including H7N7 and H7N2 AIV. In the present study, a subtype H3N2 AIV was isolated from ducks from Anhui Province, China. Sequence and phylogenetic analyses revealed that seven gene segments of this virus showed the highest sequence homology with that of the H7 subtype influenza virus, which is presumed to be the reassortants of the H3 and H7 subtypes AIV. The present study also reconfirmed that the reassortment between the H7 subtype and waterfowl-originating AIVs universally occurred in waterfowl. Animal inoculation tests showed that the virus has low pathogenicity in chickens; however, it could be replicated in the lungs of mice. The emergence of this H3N2 isolate emphasizes the importance of enhancing the surveillance of waterfowl-originating AIVs, the identification of novel reassortant strains, and characterization of their biological properties. PMID:27000112

  15. Newcastle Disease Viruses Causing Recent Outbreaks Worldwide Show Unexpectedly High Genetic Similarity to Historical Virulent Isolates from the 1940s.

    PubMed

    Dimitrov, Kiril M; Lee, Dong-Hun; Williams-Coplin, Dawn; Olivier, Timothy L; Miller, Patti J; Afonso, Claudio L

    2016-05-01

    Virulent strains of Newcastle disease virus (NDV) cause Newcastle disease (ND), a devastating disease of poultry and wild birds. Phylogenetic analyses clearly distinguish historical isolates (obtained prior to 1960) from currently circulating viruses of class II genotypes V, VI, VII, and XII through XVIII. Here, partial and complete genomic sequences of recent virulent isolates of genotypes II and IX from China, Egypt, and India were found to be nearly identical to those of historical viruses isolated in the 1940s. Phylogenetic analysis, nucleotide distances, and rates of change demonstrate that these recent isolates have not evolved significantly from the most closely related ancestors from the 1940s. The low rates of change for these virulent viruses (7.05 × 10(-5) and 2.05 × 10(-5) per year, respectively) and the minimal genetic distances existing between these and historical viruses (0.3 to 1.2%) of the same genotypes indicate an unnatural origin. As with any other RNA virus, Newcastle disease virus is expected to evolve naturally; thus, these findings suggest that some recent field isolates should be excluded from evolutionary studies. Furthermore, phylogenetic analyses show that these recent virulent isolates are more closely related to virulent strains isolated during the 1940s, which have been and continue to be used in laboratory and experimental challenge studies. Since the preservation of viable viruses in the environment for over 6 decades is highly unlikely, it is possible that the source of some of the recent virulent viruses isolated from poultry and wild birds might be laboratory viruses. PMID:26888902

  16. Development, Characterization and Application of Monoclonal Antibodies against Brazilian Dengue Virus Isolates

    PubMed Central

    Bordignon, Juliano; Duarte dos Santos, Claudia Nunes

    2014-01-01

    Dengue is the most prevalent human arboviral disease. The morbidity related to dengue infection supports the need for an early, quick and effective diagnostic test. Brazil is a hotspot for dengue, but no serological diagnostic test has been produced using Brazilian dengue virus isolates. This study aims to improve the development of immunodiagnostic methods for dengue virus (DENV) detection through the production and characterization of 22 monoclonal antibodies (mAbs) against Brazilian isolates of DENV-1, -2 and -3. The mAbs include IgG2bκ, IgG2aκ and IgG1κ isotypes, and most were raised against the envelope or the pre-membrane proteins of DENV. When the antibodies were tested against the four DENV serotypes, different reactivity patterns were identified: group-specific, subcomplex specific (DENV-1, -3 and -4 and DENV-2 and -3) and dengue serotype-specific (DENV-2 or -3). Additionally, some mAbs cross-reacted with yellow fever virus (YFV), West Nile virus (WNV) and Saint Louis encephalitis virus (SLEV). None of the mAbs recognized the alphavirus Venezuelan equine encephalitis virus (VEEV). Furthermore, mAbs D3 424/8G, D1 606/A12/B9 and D1 695/12C/2H were used to develop a capture enzyme-linked immunosorbent assay (ELISA) for anti-dengue IgM detection in sera from patients with acute dengue. To our knowledge, these are the first monoclonal antibodies raised against Brazilian DENV isolates, and they may be of special interest in the development of diagnostic assays, as well as for basic research. PMID:25412181

  17. Development, characterization and application of monoclonal antibodies against Brazilian Dengue virus isolates.

    PubMed

    Zanluca, Camila; Mazzarotto, Giovanny Augusto Camacho Antevere; Bordignon, Juliano; Duarte Dos Santos, Claudia Nunes

    2014-01-01

    Dengue is the most prevalent human arboviral disease. The morbidity related to dengue infection supports the need for an early, quick and effective diagnostic test. Brazil is a hotspot for dengue, but no serological diagnostic test has been produced using Brazilian dengue virus isolates. This study aims to improve the development of immunodiagnostic methods for dengue virus (DENV) detection through the production and characterization of 22 monoclonal antibodies (mAbs) against Brazilian isolates of DENV-1, -2 and -3. The mAbs include IgG2bκ, IgG2aκ and IgG1κ isotypes, and most were raised against the envelope or the pre-membrane proteins of DENV. When the antibodies were tested against the four DENV serotypes, different reactivity patterns were identified: group-specific, subcomplex specific (DENV-1, -3 and -4 and DENV-2 and -3) and dengue serotype-specific (DENV-2 or -3). Additionally, some mAbs cross-reacted with yellow fever virus (YFV), West Nile virus (WNV) and Saint Louis encephalitis virus (SLEV). None of the mAbs recognized the alphavirus Venezuelan equine encephalitis virus (VEEV). Furthermore, mAbs D3 424/8G, D1 606/A12/B9 and D1 695/12C/2H were used to develop a capture enzyme-linked immunosorbent assay (ELISA) for anti-dengue IgM detection in sera from patients with acute dengue. To our knowledge, these are the first monoclonal antibodies raised against Brazilian DENV isolates, and they may be of special interest in the development of diagnostic assays, as well as for basic research. PMID:25412181

  18. Genetic diversity of the coat protein of olive latent virus 1 isolates.

    PubMed

    Varanda, C M R; Nolasco, G; Clara, M I; Félix, M R

    2014-06-01

    The CP gene variability among 21 olive latent virus 1 (OLV-1) isolates obtained from different hosts and locations and at different times was assessed. Amplicons obtained by RT-PCR were cloned, and at least 10 sequences from each isolate were analyzed and compared. OLV-1 sequences available in GenBank were included. The encoded CPs consisted of 270 amino acids, except those of isolates G1S and C7 (269 aa) and G6 (271 aa). Comparison of CP genomic sequences of the isolates under study showed very low values of nucleotide diversity, 0.02, and maximum nucleotide distances between (0.087) or within isolates (0.001). Although very few nucleotide sequence differences were observed among the isolates, olive isolates exhibited lower diversity (0.012). In addition, at position 158 (157 in C7 and G1S and 159 in G6) of the deduced aa sequences, an alanine residue was found to be conserved among the olive isolates. In citrus and tulip isolates, a threonine residue was present at position 158, whereas a valine was present at this same position in tomato isolates. Phylogenetic analysis indicated that OLV-1 isolates clustered in five groups according to original host. However, G6, originally recovered from olive but repeatedly inoculated and maintained in N. benthamiana plants for 8 years in our laboratory, was separated from other isolates. This may be attributable to adaptation to the experimental host over time. There was no correlation of phylogenetic grouping of isolates based on geographical location or year of collection. Strong negative selection may have contributed to the low diversity among the OLV-1 CP isolates. PMID:24352437

  19. Prevalence and genetic diversity of fig mosaic virus isolates infecting fig tree in Iran.

    PubMed

    Danesh-Amuz, S; Rakhshandehroo, F; Rezaee, S

    2014-01-01

    Commercial and outdoor fig orchards in four Iranian provinces were surveyed for the incidence of fig mosaic virus (FMV), fig leaf mottle associated virus 2 (FLMaV-2) and fig mild mottle associated virus (FMMaV) from March 2011 to October 2012. A total of 350 asymptomatic and symptomatic fig samples were collected and tested by dot-immunobinding assay (DIBA) for the fig mosaic disease (FMD) using a polyclonal antiserum. According to DIBA results, FMD was present in 73% of the collected symptomatic samples from all visited regions. Samples with positive reactions in DIBA were then analyzed by RT-PCR using with specific primers. PCR results showed that about 14.8% of the FMD-positive samples from three inspected provinces are infected with at least one virus. FMV was the most widely spread virus (14%) followed by FLMaV-2 (1.5%), whereas FMMaV was not found. Phylogenetic analysis of the glycoprotein nucleotide and amino acid sequences of known FMV isolates showed two independent groups with high bootstrap values, with all Iranian isolates distinctly clustered in group I, subgroup IA beside those reported in Turkey. Nucleotide diversity was high within but low between different selected geographic regions and except for Europe, nucleotide distance within geographic regions was low. Statistical analyses indicated a correlation between the genetic structure of the FMV isolates and the geographical origin of isolation. Our analyses suggested that the FMV population is in a state of increase following a bottleneck or founder event in Iran. PMID:25283859

  20. Sequence comparisons of the variable VP2 region of eight infectious bursal disease virus isolates.

    PubMed

    Dormitorio, T V; Giambrone, J J; Duck, L W

    1997-01-01

    The VP2 gene is part of the genomic segment A of infectious bursal disease virus (IBDV). It has been identified as the major host-protective antigen of IBDV and is known to contain conformationally dependent protective epitopes. A 643-base pair segment covering the hypervariable region of this gene from three recent serologic variant IBDV isolates from the southeastern United States, two variants from the Delmarva Peninsula, and three serologic standard viruses were amplified and sequenced using the reverse transcription polymerase chain reaction and cycle sequencing techniques. This was done to determine the molecular similarity among isolates that differ antigenically and pathologically. Sequence analysis suggested that the Arkansas (Ark) and Mississippi (Miss) isolates evolved closely and separately from the Delmarva variants (GLS and DELE), in contrast to the other southeastern variant Georgia (Ga), which is more closely related (98.32%) to Delaware E (DELE). All variants, except for Miss, underwent a shift in amino acid number 222 from proline to threonine. The sequence of Univax BD virus, a commercially available intermediate vaccine, was markedly different, evolving from a separate lineage than the others. Restriction enzyme sites could differentiate most isolates. Except for Miss, variants do not have EcoRII site at the larger hydrophilic domain. All variants lost their HaeIII, StuI, and StyI cutting sites with a change in base number 856. The TaqI site is in DELE, whereas the SpeI site is absent in the standard vaccine viruses. The SWASASGS heptapeptide is conserved in all virulent viruses, including APHIS, but not in the attenuated (Univax BD and Bursa Vac 3) and published (D78 and PBG98) vaccines. PMID:9087318

  1. Co-culture: A quick approach for isolation of street rabies virus in murine neuroblastoma cells

    PubMed Central

    Sasikalaveni, A.; Tirumurugaan, K. G.; Manoharan, S.; Raj, G. Dhinakar; Kumanan, K.

    2015-01-01

    Background: Laboratory detection of rabies in most cases is based on detection of the antigen by fluorescent antibody test, however, in weak positive cases confirmative laboratory diagnosis depends on widely accepted mouse inoculation test. Cell lines like neuroblastoma have been used to isolate the virus with greater success not only to target for diagnosis, but also for molecular studies that determine the epidemiology of the circulating street rabies strains and in studies that look at the efficiency of the developed monoclonal antibodies to neutralize the different rabies strains. Due to the recent issues in obtaining ethical permission for mouse experimentation, and also the passages required in the cell lines to isolate the virus, we report herewith a co-culture protocol using the murine neuroblastoma (MNA) cells, which enable quicker isolation of street rabies virus with minimum passages. Objective: This study is not to have an alternative diagnostic assay, but an approach to produce sufficient amount of rabies virus in minimum passages by a co-culture approach in MNA cells. Materials and Methods: The MNA cells are co-cultured by topping the normal cells with infected cells every 48 h and the infectivity was followed up by performing direct fluorescent-antibody test. Results: The co-culture approach results in 100% infectivity and hence the use of live mouse for experimentation could be avoided. Conclusion: Co-culture method provides an alternative for the situations with limited sample volume and for the quicker isolation of virus which warrants the wild type strains without much modification. PMID:27047148

  2. Isolation of vesicular stomatitis virus defective interfering genomes with different amounts of 5'-terminal complementarity.

    PubMed Central

    Kolakofsky, D

    1982-01-01

    I isolated at least 30 different vesicular stomatitis virus defective interfering (DI) genomes, distinguished by chain length, by five independent undiluted passages of a repeatedly cloned virus plaque. Labeling of the 3' hydroxyl ends of these DI genomes and RNase digestion studies demonstrated that the ends of these DI genomes were terminally complementary to different extents (approximately 46 to 200 nucleotides). Mapping studies showed that the complementary ends of all of the DI genomes were derived from the 5' ends of the nondefective minus-strand genome. Regardless of the extent of terminal complementarity, all of the DI genomes synthesized the same 46-nucleotide minus-strand leader RNA. Images PMID:6281468

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

    PubMed Central

    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

    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

  4. Isolation and genetic characterization of avian influenza viruses and a Newcastle disease virus from wild birds in Barbados: 2003-2004.

    PubMed

    Douglas, Kirk O; Lavoie, Marc C; Kim, L Mia; Afonso, Claudio L; Suarez, David L

    2007-09-01

    Zoonotic transmission of an H5N1 avian influenza A virus to humans in 2003-present has generated increased public health and scientific interest in the prevalence and variability of influenza A viruses in wild birds and their potential threat to human health. Migratory waterfowl and shorebirds are regarded as the primordial reservoir of all influenza A viral subtypes and have been repeatedly implicated in avian influenza outbreaks in domestic poultry and swine. All of the 16 hemagglutinin and nine neuraminidase influenza subtypes have been isolated from wild birds, but waterfowl of the order Anseriformes are the most commonly infected. Using 9-to-11-day-old embryonating chicken egg culture, virus isolation attempts were conducted on 168 cloacal swabs from various resident, imported, and migratory bird species in Barbados during the months of July to October of 2003 and 2004. Hemagglutination assay and reverse transcription-polymerase chain reaction were used to screen all allantoic fluids for the presence of hemagglutinating agents and influenza A virus. Hemagglutination positive-influenza negative samples were also tested for Newcastle disease virus (NDV), which is also found in waterfowl. Two influenza A viruses and one NDV were isolated from Anseriformes (40/168), with isolation rates of 5.0% (2/40) and 2.5% (1/40), respectively, for influenza A and NDV. Sequence analysis of the influenza A virus isolates showed them to be H4N3 viruses that clustered with other North American avian influenza viruses. This is the first report of the presence of influenza A virus and NDV in wild birds in the English-speaking Caribbean. PMID:17992942

  5. Evolutionary and phenotypic analysis of live virus isolates suggests arthropod origin of a pathogenic RNA virus family.

    PubMed

    Marklewitz, Marco; Zirkel, Florian; Kurth, Andreas; Drosten, Christian; Junglen, Sandra

    2015-06-16

    The evolutionary origins of arboviruses are unknown because their typical dual host tropism is paraphyletic within viral families. Here we studied one of the most diversified and medically relevant RNA virus families, the Bunyaviridae, in which four of five established genera are transmitted by arthropods. We define two cardinally novel bunyavirus groups based on live isolation of 26 viral strains from mosquitoes (Jonchet virus [JONV], eight strains; Ferak virus [FERV], 18 strains). Both viruses were incapable of replicating at vertebrate-typical temperatures but replicated efficiently in insect cells. Replication involved formation of virion-sense RNA (vRNA) and mRNA, including cap-snatching activity. SDS/PAGE, mass spectrometry, and Edman degradation identified translation products corresponding to virion-associated RNA-dependent RNA polymerase protein (RdRp), glycoprotein precursor protein, glycoproteins Gn and Gc, as well as putative nonstructural proteins NSs and NSm. Distinct virion morphologies suggested ancient evolutionary divergence, with bunyavirus-typical morphology for FERV (spheres of 60-120 nm) as opposed to an unusual bimorphology for JONV (tubular virions of 60 × 600 nm and spheres of 80 nm). Both viruses were genetically equidistant from all other bunyaviruses, showing <15% amino acid identity in the RdRp palm domain. Both had different and unique conserved genome termini, as in separate bunyavirus genera. JONV and FERV define two novel sister taxons to the superclade of orthobunyaviruses, tospoviruses, and hantaviruses. Phylogenetic ancestral state reconstruction with probabilistic hypothesis testing suggested ancestral associations with arthropods at deep nodes throughout the bunyavirus tree. Our findings suggest an arthropod origin of bunyaviruses. PMID:26038576

  6. Evolutionary and phenotypic analysis of live virus isolates suggests arthropod origin of a pathogenic RNA virus family

    PubMed Central

    Marklewitz, Marco; Zirkel, Florian; Kurth, Andreas; Drosten, Christian; Junglen, Sandra

    2015-01-01

    The evolutionary origins of arboviruses are unknown because their typical dual host tropism is paraphyletic within viral families. Here we studied one of the most diversified and medically relevant RNA virus families, the Bunyaviridae, in which four of five established genera are transmitted by arthropods. We define two cardinally novel bunyavirus groups based on live isolation of 26 viral strains from mosquitoes (Jonchet virus [JONV], eight strains; Ferak virus [FERV], 18 strains). Both viruses were incapable of replicating at vertebrate-typical temperatures but replicated efficiently in insect cells. Replication involved formation of virion-sense RNA (vRNA) and mRNA, including cap-snatching activity. SDS/PAGE, mass spectrometry, and Edman degradation identified translation products corresponding to virion-associated RNA-dependent RNA polymerase protein (RdRp), glycoprotein precursor protein, glycoproteins Gn and Gc, as well as putative nonstructural proteins NSs and NSm. Distinct virion morphologies suggested ancient evolutionary divergence, with bunyavirus-typical morphology for FERV (spheres of 60–120 nm) as opposed to an unusual bimorphology for JONV (tubular virions of 60 × 600 nm and spheres of 80 nm). Both viruses were genetically equidistant from all other bunyaviruses, showing <15% amino acid identity in the RdRp palm domain. Both had different and unique conserved genome termini, as in separate bunyavirus genera. JONV and FERV define two novel sister taxons to the superclade of orthobunyaviruses, tospoviruses, and hantaviruses. Phylogenetic ancestral state reconstruction with probabilistic hypothesis testing suggested ancestral associations with arthropods at deep nodes throughout the bunyavirus tree. Our findings suggest an arthropod origin of bunyaviruses. PMID:26038576

  7. Zinc Salts Inactivate Clinical Isolates of Herpes Simplex Virus In Vitro

    PubMed Central

    Arens, Max; Travis, Sharon

    2000-01-01

    Using a standard plaque assay and clinical isolates of herpes simplex virus (HSV), we have tested the ability of zinc salts to inactivate HSV. Virus was treated by incubation at 37°C with zinc salts in morpholinepropanesulfonic acid-buffered culture medium and was then diluted and plated onto CV-1 cells for detection and quantitation of remaining infectious virus. Of 10 randomly chosen clinical isolates (five HSV type 1 [HSV-1] isolates and five HSV-2 isolates), seven were inactivated >98% by treatment in vitro with 50 mM zinc gluconate for 2 h and nine were inactivated >97% by treatment with zinc lactate. The effect was concentration dependent. With an HSV-1 isolate, 50 mM zinc gluconate or zinc lactate caused 100% inactivation, 15 mM caused 98 to 99% inactivation, and 5 mM caused 63 to 86% inactivation. With an HSV-2 isolate, 50 and 15 mM zinc gluconate caused 30% inactivation and 5 and 1 mM caused less than 9% inactivation, whereas 50 and 15 mM zinc lactate caused greater than 92% inactivation and 5 and 1 mM caused 37 and 26% inactivation, respectively. The ability of the zinc salts to inactivate HSV was not related to pH in the pH range of 6.1 to 7.6 since inactivation by zinc gluconate or zinc lactate in that pH range was 99.7 to 100% with a 2-h treatment with 50 mM zinc salt. Short (5-min) treatments of selected isolates with zinc gluconate, zinc lactate, zinc acetate, or zinc sulfate yielded inactivation rates of 0 to 55%. PMID:10790094

  8. Phylogenetic analysis of Dengue virus 1 isolated from South Minas Gerais, Brazil

    PubMed Central

    Drumond, Betania Paiva; da Silva Fagundes, Luiz Gustavo; Rocha, Raissa Prado; Fumagalli, Marcilio Jorge; Araki, Carlos Shigueru; Colombo, Tatiana Elisa; Nogueira, Mauricio Lacerda; Castilho, Thiago Elias; da Silveira, Nelson José Freitas; Malaquias, Luiz Cosme Cotta; Coelho, Luiz Felipe Leomil

    2016-01-01

    Dengue is a major worldwide public health problem, especially in the tropical and subtropical regions of the world. Primary infection with a single Dengue virus serotype causes a mild, self-limiting febrile illness called dengue fever. However, a subset of patients who experience secondary infection with a different serotype can progress to a more severe form of the disease, called dengue hemorrhagic fever. The four Dengue virus serotypes (1–4) are antigenically and genetically distinct and each serotype is composed of multiple genotypes. In this study we isolated one Dengue virus 1 serotype, named BR/Alfenas/2012, from a patient with dengue hemorrhagic fever in Alfenas, South Minas Gerais, Brazil and molecular identification was performed based on the analysis of NS5 gene. Swiss mice were infected with this isolate to verify its potential to induce histopathological alterations characteristic of dengue. Liver histopathological analysis of infected animals showed the presence of inflammatory infiltrates, hepatic steatosis, as well as edema, hemorrhage and necrosis focal points. Phylogenetic and evolutionary analyses based on the envelope gene provided evidence that the isolate BR/Alfenas/2012 belongs to genotype V, lineage I and it is probably derived from isolates of Rio de Janeiro, Brazil. The isolate BR/Alfenas/2012 showed two unique amino acids substitutions (SER222THRE and PHE306SER) when compared to other Brazilian isolates from the same genotype/lineage. Molecular models were generated for the envelope protein indicating that the amino acid alteration PHE 306 SER could contribute to a different folding in this region located within the domain III. Further genetic and animal model studies using BR/Alfenas/2012 and other isolates belonging to the same lineage/genotype could help determine the relation of these genetic alterations and dengue hemorrhagic fever in a susceptible population. PMID:26887252

  9. Phylogenetic analysis of Dengue virus 1 isolated from South Minas Gerais, Brazil.

    PubMed

    Drumond, Betania Paiva; Fagundes, Luiz Gustavo da Silva; Rocha, Raissa Prado; Fumagalli, Marcilio Jorge; Araki, Carlos Shigueru; Colombo, Tatiana Elisa; Nogueira, Mauricio Lacerda; Castilho, Thiago Elias; da Silveira, Nelson José Freitas; Malaquias, Luiz Cosme Cotta; Coelho, Luiz Felipe Leomil

    2016-01-01

    Dengue is a major worldwide public health problem, especially in the tropical and subtropical regions of the world. Primary infection with a single Dengue virus serotype causes a mild, self-limiting febrile illness called dengue fever. However, a subset of patients who experience secondary infection with a different serotype can progress to a more severe form of the disease, called dengue hemorrhagic fever. The four Dengue virus serotypes (1-4) are antigenically and genetically distinct and each serotype is composed of multiple genotypes. In this study we isolated one Dengue virus 1 serotype, named BR/Alfenas/2012, from a patient with dengue hemorrhagic fever in Alfenas, South Minas Gerais, Brazil and molecular identification was performed based on the analysis of NS5 gene. Swiss mice were infected with this isolate to verify its potential to induce histopathological alterations characteristic of dengue. Liver histopathological analysis of infected animals showed the presence of inflammatory infiltrates, hepatic steatosis, as well as edema, hemorrhage and necrosis focal points. Phylogenetic and evolutionary analyses based on the envelope gene provided evidence that the isolate BR/Alfenas/2012 belongs to genotype V, lineage I and it is probably derived from isolates of Rio de Janeiro, Brazil. The isolate BR/Alfenas/2012 showed two unique amino acids substitutions (SER222THRE and PHE306SER) when compared to other Brazilian isolates from the same genotype/lineage. Molecular models were generated for the envelope protein indicating that the amino acid alteration PHE 306 SER could contribute to a different folding in this region located within the domain III. Further genetic and animal model studies using BR/Alfenas/2012 and other isolates belonging to the same lineage/genotype could help determine the relation of these genetic alterations and dengue hemorrhagic fever in a susceptible population. PMID:26887252

  10. CHARACTERIZATION OF VARIOUS ISOLATES OF A NATURALLY OCCURRING RECOMBINANT AVIAN LEUKOSIS VIRUS USING BIOLOGICAL ASSAYS AND POLYMERASE CHAIN REACTION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recently, we have isolated a naturally occurring recombinant avian leukosis virus (ALV) containing the envelope of ALV-B and LTR of ALV-J from commercial layer flocks affected with myeloid leukosis. Seven new isolates of the recombinant ALV, isolated from the same flock, were characterized using bio...

  11. Genotype diversity of H9N2 viruses isolated from wild birds and chickens in Hunan Province, China.

    PubMed

    Wang, Ba; Liu, Zhihua; Chen, Quanjiao; Gao, Zhimin; Fang, Fang; Chang, Haiyan; Chen, Jianjun; Xu, Bing; Chen, Ze

    2014-01-01

    Three H9N2 avian influenza viruses were isolated from the Dongting Lake wetland, among which one was from fresh egret feces, the other two were from chicken cloacal swabs in poultry markets. Phylogenetic analyses suggested that eight genes of the egret-derived H9N2 virus might come from Korean-like or American-like lineages. The two poultry-derived H9N2 viruses were reassortants between the CK/BJ/94-like and G1-like viruses. Except the PB1 genes (90.6%), the nucleotide sequence of other internal genes of the two viruses exhibited high homology (>95%). In addition, they also exhibited high homology (96-98.3%) with some genes of the H7N9 virus that caused an epidemic in China in 2013. Nucleotide sequence of the poultry-derived and egret-derived H9N2 viruses shared low homology. Infection studies showed that the egret-derived H9N2 virus was non-pathogenic to both mice and chickens, and the virus was unable to infect chickens even through 8 passages continuously in the lung. On the other hand, the chickens infected by poultry-derived viruses showed obvious clinical symptoms and even died; the infected mice showed no noticeable clinical symptoms and weight loss, but viruses could be detected in their lungs. In conclusion, for the egret-derived H9N2 virus, it would take a long adaptation process to achieve cross-species transmission in poultry and mammals. H9N2 viruses isolated at different times from the same host species in the same geographical region presented different evolutionary status, and virus isolated from different hosts in the same geographical region exhibited genetic diversity. Therefore, it is important to continue the H9N2 virus surveillance for understanding their evolutionary trends so as to provide guidance for disease control and prevention. PMID:24979703

  12. Virion envelope glycoproteins as epidermiological markers of Venezuelan encephalitis virus isolates.

    PubMed Central

    Wiebe, M E; Scherer, W F

    1980-01-01

    Virion polypeptide compositions of 26 isolates of Venezuelan encephalitis virus were analyzed by a reproducible and comparative technique of discontinuous sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis. Although the molecular weight of the core polypeptide for each isolate was 36,000, numbers and molecular weights of envelope glycoproteins were heterogeneous. Isolates associated with human, but not equine, disease usually had two glycoproteins of 50,000 to 51,000 and 51,000 to 55,000 molecular weight, whereas isolates associated with both human and equine disease usually had an additional, third polypeptide band of either 45,000 to 46,000 or 56,000 to 58,000 molecular weight. The former isolates were in hemagglutination inhibition subtypes I-D, I-E, III, or IV, and the latter were in subtypes I-A, I-B, I-C, or II. Thus virion envelope glycoproteins should be useful markers of Venezuelan encephalitis virus isolates in epidemiological investigations. PMID:7372798

  13. Genetic characteristics of mumps viruses isolated in Korea from 2007 to 2012.

    PubMed

    Kim, Seung Tae; Kim, You-Jin; Yang, Jeong-Sun; Nam, Jeong-Gu; Kim, Kisoon; Kim, Sung Soon; Kang, Hae Ji

    2016-09-01

    Mumps is a vaccine-preventable viral disease. Despite vaccine coverage of >95%, the incidence of mumps has increased in Korea since 2007. This study aimed to genetically characterize mumps virus (MuV) strains that circulated in Korea between 2007 and 2012 to determine the factors underlying mumps outbreaks. MuV was isolated from 175 clinical specimens between 2007 and 2012 in Korea. Upon analysis of the SH gene in Korean mumps virus isolates, three different genotypes were identified: I, H, and F. The MuV genotypes I and H co-circulated in Korea, and eight isolates of Korean genotype F were found within the same time period in 2008. An analysis of HN amino-acid sequence data showed that Korean isolates had no changes in their glycosylation sites. At putative neutralizing epitope sites, the Jeryl-Lynn strain showed 4-5 different amino acid sequences from those observed in Korean isolates. Korean isolates of genotypes I and H shared distinctive point mutations on putative neutralizing epitope positions in each genotype. This report describes the genetic characteristics of MuV strains circulating in Korea and provides information on endemic mumps infections. This information may be important to help prevent mumps and control outbreaks of mumps in Korea. J. Med. Virol. 88:1479-1486, 2016. © 2016 Wiley Periodicals, Inc. PMID:26950767

  14. Characterization of a Borna disease virus field isolate which shows efficient viral propagation and transmissibility.

    PubMed

    Watanabe, Yohei; Ibrahim, Madiha S; Hagiwara, Katsuro; Okamoto, Minoru; Kamitani, Wataru; Yanai, Hideyuki; Ohtaki, Naohiro; Hayashi, Yohei; Taniyama, Hiroyuki; Ikuta, Kazuyoshi; Tomonaga, Keizo

    2007-04-01

    To investigate the biological characteristics of field isolates of Borna disease virus (BDV), as well as to understand BDV infections outside endemic countries, we isolated the virus from brain samples of a heifer with Borna disease in Japan. We demonstrate that the brain lysate contained replication products of BDV and induced viral propagation in rat glioma cells, suggesting that a replication-competent BDV existed in the bovine brain. This field strain of BDV, named Bo/04w, showed efficient viral release and transmissibility and also displayed a distinct pattern of expression of viral phosphoprotein (P) during infection, as compared with laboratory-adapted BDV strains. Interestingly, we found the level of P to be significantly low in cells infected with Bo/04w, and the transcription of this isolate to be more efficient than that of laboratory strain of BDV. These results indicated that the field isolate may regulate the expression of P at an optimal level in infected cells. We also confirmed that Bo/04w maintains biological significance in neonatal gerbil brain. Sequencing revealed that despite the biological differences, the field isolate is closely related genetically to the laboratory strains of BDV. We discuss here the sequence similarities between BDV isolates from endemic and nonendemic countries. PMID:17306587

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

    PubMed Central

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

    2012-01-01

    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

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

    PubMed Central

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

    1978-01-01

    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

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

    SciTech Connect

    Lamb, Kristen; Lokesh, G.L.; Sherman, Michael; Watowich, Stanley

    2010-10-25

    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.

  18. Respiratory infectivity of a recently isolated Egyptian strain of Rift Valley fever virus.

    PubMed Central

    Brown, J L; Dominik, J W; Morrissey, R L

    1981-01-01

    The respiratory infectivity of a strain of Rift Valley fever virus isolated in Egypt (strain ZH-501) was compared with that of one isolate from Uganda (Entebbe strain) and two isolates from South Africa (strains SA-51 and SA-75). Studies were performed with ICR mice which were infected by exposure to infectious aerosols composed of particles with a mass median diameter of 0.96 micrometer. The respiratory median lethal doses for ZH-501, Entebbe, SA-51, and SA-75 were 2.2, 1.9, 2.6, and 1.9 log10 plaque-forming units, respectively. Although these values are statistically different, the biological implications of such differences seem unimportant. In an additional study of pathogenesis, a single group of mice was infected with 3.1 log10 plaque-forming units of ZH-501, and tissues were assayed sequentially through 96 h postinfection. Between 6 and 30 h, demonstration of an increasing virus concentration only in the lungs indicated that initial replication occurred there; however, determination of histopathological changes did not reveal evidence of pneumonia. Virus was isolated from the liver by 48 h, and the ultimate outcome of infection was a fulminating and fatal hepatic necrosis. PMID:7287187

  19. FMD virus isolates: the candidate strains for polyvalent vaccine development in Ethiopia.

    PubMed

    Ayelet, G; Soressa, M; Sisay, T; Belay, A; Gelaye, E; Jembere, S; Skjerve, E; Asmare, K

    2013-06-01

    The study was conducted on foot-and-mouth disease (FMD) viruses with the aim of selecting appropriate vaccinal strain to control of FMD in Ethiopia. The study was conducted in two-dimensional virus neutralization assay to determine the antigenic relationship 'r' value between the candidate vaccine strains and field isolates. A total of 21 serotype O, 7 serotype A, and 8 serotype SAT 2 FMD viruses, which were isolated from cattle and swine. A couple of isolates from each serotype were identified as vaccine candidates in the trial (O-ETH/38/2005, O-ETH/58/2008, A-ETH/7/2008, A-ETH/6/2000, SAT2-ETH/76/2009 and SAT2-ETH/64/2009). The finding revealed all the vaccine candidate depicted high antigenic similarity, above the mean "r" value, to their own serotypes in the studied serotype population except for one serotype A field isolate, A-ETH/13/1981, with "r" value=0.14 and 0.25) which is significantly lower than the minimum requirement. In general, the result indicated that these candidate vaccinal strains can be used for polyvalent vaccine production in the country. PMID:23416124

  20. Phylogenetic analysis of Indian rabies virus isolates targeting the complete glycoprotein gene.

    PubMed

    Cherian, Susan; Singh, Rajendra; Singh, K P; Manjunatha Reddy, G B; Anjaneya; Ravi Kumar, G V P P S; Sumithra, T G; Singh, R P

    2015-12-01

    Rabies a fatal viral zoonosis is endemic in India. There is no report on phylogenetic study of Indian rabies virus isolates based on the complete G gene. In the present study, a total of 25 rabies positive brain samples collected during 2001-2014 from North India (UP, MP, Delhi, Rajasthan), South India (Kerala and Karnataka) and Gujarat states belonging to six different host species were subjected to G gene amplification by RT-PCR as three overlapping fragments of 881 bp, 991 bp and 618 bp. Phylogenetic analysis revealed that all Indian rabies virus isolates are genetically closely related with Arctic-like 1a lineage viruses. However, two distinct clusters were identified namely, India South and India North. All the Indian rabies isolates had 95.5-100% homology related to geography, but not to host species. Deduced amino acids on comparison revealed two amino acid changes, aa 356 in ECTO; N→K and aa 458; M→I, which were found to distinguish between the India South and India North isolates. PMID:26427850

  1. Genome sequence analysis of dengue virus 1 isolated in Key West, Florida.

    PubMed

    Shin, Dongyoung; Richards, Stephanie L; Alto, Barry W; Bettinardi, David J; Smartt, Chelsea T

    2013-01-01

    Dengue virus (DENV) is transmitted to humans through the bite of mosquitoes. In November 2010, a dengue outbreak was reported in Monroe County in southern Florida (FL), including greater than 20 confirmed human cases. The virus collected from the human cases was verified as DENV serotype 1 (DENV-1) and one isolate was provided for sequence analysis. RNA was extracted from the DENV-1 isolate and was used in reverse transcription polymerase chain reaction (RT-PCR) to amplify PCR fragments to sequence. Nucleic acid primers were designed to generate overlapping PCR fragments that covered the entire genome. The DENV-1 isolate found in Key West (KW), FL was sequenced for whole genome characterization. Sequence assembly, Genbank searches, and recombination analyses were performed to verify the identity of the genome sequences and to determine percent similarity to known DENV-1 sequences. We show that the KW DENV-1 strain is 99% identical to Nicaraguan and Mexican DENV-1 strains. Phylogenetic and recombination analyses suggest that the DENV-1 isolated in KW originated from Nicaragua (NI) and the KW strain may circulate in KW. Also, recombination analysis results detected recombination events in the KW strain compared to DENV-1 strains from Puerto Rico. We evaluate the relative growth of KW strain of DENV-1 compared to other dengue viruses to determine whether the underlying genetics of the strain is associated with a replicative advantage, an important consideration since local transmission of DENV may result because domestic tourism can spread DENVs. PMID:24098658

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

    USGS Publications Warehouse

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

    2009-01-01

    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.

  3. High-Throughput Isolation of Giant Viruses in Liquid Medium Using Automated Flow Cytometry and Fluorescence Staining

    PubMed Central

    Khalil, Jacques Y. B.; Robert, Stephane; Reteno, Dorine G.; Andreani, Julien; Raoult, Didier; La Scola, Bernard

    2016-01-01

    The isolation of giant viruses using amoeba co-culture is tedious and fastidious. Recently, the procedure was successfully associated with a method that detects amoebal lysis on agar plates. However, the procedure remains time-consuming and is limited to protozoa growing on agar. We present here advances for the isolation of giant viruses. A high-throughput automated method based on flow cytometry and fluorescent staining was used to detect the presence of giant viruses in liquid medium. Development was carried out with the Acanthamoeba polyphaga strain widely used in past and current co-culture experiments. The proof of concept was validated with virus suspensions: artificially contaminated samples but also environmental samples from which viruses were previously isolated. After validating the technique, and fortuitously isolating a new Mimivirus, we automated the technique on 96-well plates and tested it on clinical and environmental samples using other protozoa. This allowed us to detect more than 10 strains of previously known species of giant viruses and seven new strains of a new virus lineage. This automated high-throughput method demonstrated significant time saving, and higher sensitivity than older techniques. It thus creates the means to isolate giant viruses at high speed. PMID:26858703

  4. High-Throughput Isolation of Giant Viruses in Liquid Medium Using Automated Flow Cytometry and Fluorescence Staining.

    PubMed

    Khalil, Jacques Y B; Robert, Stephane; Reteno, Dorine G; Andreani, Julien; Raoult, Didier; La Scola, Bernard

    2016-01-01

    The isolation of giant viruses using amoeba co-culture is tedious and fastidious. Recently, the procedure was successfully associated with a method that detects amoebal lysis on agar plates. However, the procedure remains time-consuming and is limited to protozoa growing on agar. We present here advances for the isolation of giant viruses. A high-throughput automated method based on flow cytometry and fluorescent staining was used to detect the presence of giant viruses in liquid medium. Development was carried out with the Acanthamoeba polyphaga strain widely used in past and current co-culture experiments. The proof of concept was validated with virus suspensions: artificially contaminated samples but also environmental samples from which viruses were previously isolated. After validating the technique, and fortuitously isolating a new Mimivirus, we automated the technique on 96-well plates and tested it on clinical and environmental samples using other protozoa. This allowed us to detect more than 10 strains of previously known species of giant viruses and seven new strains of a new virus lineage. This automated high-throughput method demonstrated significant time saving, and higher sensitivity than older techniques. It thus creates the means to isolate giant viruses at high speed. PMID:26858703

  5. Isolation and characterisation of equine influenza viruses (H3N8) from Europe and North America from 2008 to 2009.

    PubMed

    Bryant, Neil A; Rash, Adam S; Woodward, Alana L; Medcalf, Elizabeth; Helwegen, Maud; Wohlfender, Franziska; Cruz, Fatima; Herrmann, Claudia; Borchers, Kerstin; Tiwari, Ashish; Chambers, Thomas M; Newton, J Richard; Mumford, Jennifer A; Elton, Debra M

    2011-01-10

    Like other influenza A viruses, equine influenza virus undergoes antigenic drift. It is therefore essential that surveillance is carried out to ensure that recommended strains for inclusion in vaccines are kept up to date. Here we report antigenic and genetic characterisation carried out on equine influenza virus strains isolated in North America and Europe over a 2-year period from 2008 to 2009. Nasopharyngeal swabs were taken from equines showing acute clinical signs and submitted to diagnostic laboratories for testing and virus isolation in eggs. The sequence of the HA1 portion of the viral haemagglutinin was determined for each strain. Where possible, sequence was determined directly from swab material as well as from virus isolated in eggs. In Europe, 20 viruses were isolated from 15 sporadic outbreaks and 5 viruses were isolated from North America. All of the European and North American viruses were characterised as members of the Florida sublineage, with similarity to A/eq/Lincolnshire/1/07 (clade 1) or A/eq/Richmond/1/07 (clade 2). Antigenic characterisation by haemagglutination inhibition assay indicated that the two clades could be readily distinguished and there were also at least seven amino acid differences between them. The selection of vaccine strains for 2010 by the expert surveillance panel have taken these differences into account and it is now recommended that representatives of both Florida clade 1 and clade 2 are included in vaccines. PMID:20580170

  6. Isolation and identification of highly pathogenic avian influenza virus subtype H5N1 in peafowl (Pavo cristatus).

    PubMed

    Ismail, Mahmoud Moussa; Khan, Owais Ahmed; Cattoli, Giovanni; Lu, Huaguang

    2010-03-01

    An outbreak of highly pathogenic avian influenza (HPAI) virus subtype H5N1 was first diagnosed in a "backyard" flock of peafowl (Pavo cristatus) raised on palace premises in the Kingdom of Saudi Arabia in December 3, 2007. The flock consisted of 40 peafowl, and their ages ranged from 3 to 5 years old. Affected birds suffered from depression, anorexia, and white diarrhea. Four dead birds were submitted for HPAI diagnosis at the Central Veterinary Diagnostic Laboratory in Riyadh. Brain and liver tissues and tracheal and cloacal swabs were taken from the dead birds and processed for a real-time reverse transcriptase (RT)-PCR test and virus isolation in specific-pathogen-free embryonating chicken eggs. The H5N1 subtype of avian influenza virus was isolated from the four dead birds and identified by a real-time RT-PCR before and after egg inoculation. The virus isolates were characterized as HPAI H5N1 virus by sequencing analysis. Phylogenetic comparisons revealed that the H5N1 viruses isolated from peafowl belong to the genetic clade 2.2 according to the World Health Organization nomenclature. The peafowl H5N1 virus falls into 2.2.2 sublineage II and clusters with the H5N1 viruses isolated from poultry in Saudi Arabia in 2007-08. PMID:20521659

  7. Molecular characterization of a Class I Newcastle disease virus strain isolated from a pigeon in China.

    PubMed

    Ren, Shanhui; Xie, Xiumei; Wang, Yanping; Tong, Lina; Gao, Xiaolong; Jia, Yanqing; Wang, Haixin; Fan, Mengfei; Zhang, Shuxia; Xiao, Sa; Wang, Xinglong; Yang, Zengqi

    2016-08-01

    Constant monitoring is performed to elucidate the role of natural hosts in the ecology of Newcastle disease virus (NDV). In this study, an NDV strain isolated from an asymptomatic pigeon was sequenced and analysed. Results showed that the full-length genomes of this isolate were 15,198 nucleotides with the gene order of 3'-NP-P-M-F-HN-L-5'. This NDV isolate was lentogenic, with an intracerebral pathogenicity index of 0.00 and a mean time of death more than 148 h. The isolate possessed a motif of -(112)E-R-Q-E-R-L(117)- at the F protein cleavage site. In addition, 7 and 13 amino acid substitutions were identified in the functional domains of fusion protein (F) and haemagglutinin-neuraminidase protein (HN) proteins, respectively. Analysis of the amino acids of neutralizing epitopes of F and HN proteins showed 3 and 10 amino acid substitutions, respectively, in the isolate. Phylogenetic analysis classified the isolate into genotype Ib in Class I. This isolate shared high homologies with the NDV strains isolated from wild birds and waterfowl in southern and eastern parts of China from 2005 to 2013. To our knowledge, this study is the first to report a NDV strain isolated from pigeon that belongs to genotype Ib in Class I, rather than to the traditional genotype VI or other sub-genotypes in Class II. This study provides information to elucidate the distribution and evolution of Class I viruses for further NDV prevention. PMID:26950543

  8. Isolation of a GB virus-related genome from a chimpanzee.

    PubMed

    Birkenmeyer, L G; Desai, S M; Muerhoff, A S; Leary, T P; Simons, J N; Montes, C C; Mushahwar, I K

    1998-09-01

    Recently, two new flaviviruses, GB virus A (GBV-A) and GB virus B (GBV-B), were identified in the plasma of a tamarin infected with the hepatitis GB agent. A third virus, GB virus C (GBV-C), was subsequently identified in humans. In the current study, representational difference analysis (RDA) was used to search for a new virus in the serum of a chimpanzee that developed acute resolving hepatitis following inoculation with a pool of chimpanzee plasma. The plasma pool originated from serial passages of a human sample containing virus-like particles. Numerous cDNA clones were obtained that exhibited 62-80% identity with GBV-C. With the exception of the extreme 5' and 3' ends, the complete viral genome was sequenced, revealing a single large open reading frame encoding a 2833 amino acid polyprotein that contains two envelope proteins, two proteases, a helicase, and an RNA-dependent RNA polymerase. Phylogenetic analysis of the new virus indicates that it is closely related to GBV-C, yet still sufficiently divergent as to be placed in a separate group, tentatively labeled GB virus Ctroglodytes (GBV-Ctro). Numerous human samples were screened by reverse transcriptase-polymerase chain reaction (RT-PCR), but GBV-Ctro sequence was not detected. However, a second chimpanzee inoculated with the same plasma pool was shown to develop a GBV-Ctro infection. Although isolated from an Old World primate with hepatitis, the primary host of GBV-Ctro and any association with disease remains to be determined. PMID:9700632

  9. Complete genome sequences of two biologically distinct isolates of Asparagus virus 1.

    PubMed

    Blockus, S; Lesker, T; Maiss, E

    2015-02-01

    The complete genome sequences of two asparagus virus 1 (AV-1) isolates differing in their ability to cause systemic infection in Nicotiana benthamiana were determined. Their genomes had 9,741 nucleotides excluding the 3'-terminal poly(A) tail, encoded a polyprotein of 3,112 amino acids, and shared 99.6 % nucleotide sequence identity. They differed at 37 nucleotide and 15 amino acid sequence positions (99.5 % identity) scattered over the polyprotein. The closest relatives of AV-1 in amino acid sequence identity were plum pox virus (54 %) and turnip mosaic virus (53 %), corroborating the classification of AV-1 as a member of a distinct species in the genus Potyvirus. PMID:25216774

  10. Cymbidium chlorotic mosaic virus, a new sobemovirus isolated from a spring orchid (Cymbidium goeringii) in Japan.

    PubMed

    Kondo, Hideki; Takemoto, Shogo; Maruyama, Kazuyuki; Chiba, Sotaro; Andika, Ida Bagus; Suzuki, Nobuhiro

    2015-08-01

    Cymbidium chlorotic mosaic virus (CyCMV), isolated from a spring orchid (Cymbidium goeringii), was characterized molecularly. CyCMV isometric virions comprise a single, positive-strand RNA genome of 4,083 nucleotides and 30-kDa coat protein. The virus genome contains five overlapping open reading frames with a genomic organization similar to that of sobemoviruses. BLAST searches and phylogenetic analysis revealed that CyCMV is most closely related to papaya lethal yellowing virus, a proposed dicot-infecting sobemovirus (58.8 % nucleotide sequence identity), but has a relatively distant relationship to monocot-infecting sobemoviruses, with only modest sequence identities. This suggests that CyCMV is a new monocot-infecting member of the floating genus Sobemovirus. PMID:26025156

  11. Isolation and Physiological Characterization of a Novel Algicidal Virus Infecting the Marine Diatom Skeletonema costatum

    PubMed Central

    Kim, JinJoo; Kim, Chang-Hoon; Youn, Seok-Hyun; Choi, Tae-Jin

    2015-01-01

    Diatoms are a major component of the biological community, serving as the principal primary producers in the food web and sustaining oxygen levels in aquatic environments. Among marine planktonic diatoms, the cosmopolitan Skeletonema costatum is one of the most abundant and widespread species in the world’s oceans. Here, we report the basic characteristics of a new diatom-infecting S. costatum virus (ScosV) isolated from Jaran Bay, Korea, in June 2008. ScosV is a polyhedral virus (45–50 nm in diameter) that propagates in the cytoplasm of host cells and causes lysis of S. costatum cultures. The infectivity of ScosV was determined to be strain- rather than species-specific, similar to other algal viruses. The burst size and latent period were roughly estimated at 90–250 infectious units/cell and <48 h, respectively. PMID:26060438

  12. Isolation and Physiological Characterization of a Novel Algicidal Virus Infecting the Marine Diatom Skeletonema costatum.

    PubMed

    Kim, JinJoo; Kim, Chang-Hoon; Youn, Seok-Hyun; Choi, Tae-Jin

    2015-06-01

    Diatoms are a major component of the biological community, serving as the principal primary producers in the food web and sustaining oxygen levels in aquatic environments. Among marine planktonic diatoms, the cosmopolitan Skeletonema costatum is one of the most abundant and widespread species in the world's oceans. Here, we report the basic characteristics of a new diatom-infecting S. costatum virus (ScosV) isolated from Jaran Bay, Korea, in June 2008. ScosV is a polyhedral virus (45-50 nm in diameter) that propagates in the cytoplasm of host cells and causes lysis of S. costatum cultures. The infectivity of ScosV was determined to be strain- rather than species-specific, similar to other algal viruses. The burst size and latent period were roughly estimated at 90-250 infectious units/cell and <48 h, respectively. PMID:26060438

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

    SciTech Connect

    Muggeridge, Martin I. . E-mail: mmugge@lsuhsc.edu; Grantham, Michael L.; Johnson, F. Brent

    2004-10-25

    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.

  14. Genetic variability of blueberry scorch virus isolates from highbush blueberry in New York State.

    PubMed

    Kalinowska, Elżbieta; Marsella-Herrick, Patricia; Fuchs, Marc

    2015-06-01

    The genetic variability of blueberry scorch virus (BlScV) isolates from New York was determined within a portion of the RNA-dependent RNA polymerase gene and the triple gene block and coat protein (CP) genes. Phylogenetic analysis of 19 New York isolates and other isolates for which sequence information is available in GenBank revealed two distinct clades, regardless of the coding region analyzed, and limited variability within (0.029 ± 0.007) and between (0.183 ± 0.032) phylogroups. Recombination events were identified in the CP gene of three New York isolates, and codons of the five BlScV genes characterized were found to be under neutral or negative selective pressure. PMID:25809019

  15. Synthesis of herpes simplex virus, vaccinia virus, and adenovirus DNA in isolated HeLa cell nuclei. I. Effect of viral-specific antisera and phosphonoacetic acid.

    PubMed Central

    Bolden, A; Aucker, J; Weissbach, A

    1975-01-01

    Purified nuclei, isolated from appropriately infected HeLa cells, are shown to synthesize large amounts of either herpes simplex virus (HSV) or vaccinia virus DNA in vitro. The rate of synthesis of DNA by nuclei from infected cells is up to 30 times higher than the synthesis of host DNA in vitro by nuclei isolated from uninfected HeLa cells. Thus HSV nuclei obtained from HSV-infected cells make DNA in vitro at a rate comparable to that seen in the intact, infected cell. Molecular hybridization studies showed that 80% of the DNA sequences synthesized in vitro by nuclei from herpesvirus-infected cells are herpesvirus specific. Vaccinia virus nuclei from vaccinia virus-infected cells, also produce comparable percentages of vaccinia virus-specific DNA sequences. Adenovirus nuclei from adenovirus 2-infected HeLa cells, which also synthesize viral DNA in vitro, have been included in this study. Synthesis of DNA by HSV or vaccinia virus nuclei is markedly inhibited by the corresponding viral-specific antisera. These antisera inhibit in a similar fashion the purified herpesvirus-induced or vaccinia virus-induced DNA polymerase isolated from infected cells. Phosphonoacetic acid, reported to be a specific inhibitor of herpesvirus formation and the herpesvirus-induced DNA polymerase, is equally effective as an inhibitor of HSV DNA synthesis in isolated nuclei in vitro. However, we also find phosphonoacetic acid to be an effective inhibitor of vaccinia virus nuclear DNA synthesis and the purified vaccinia virus-induced DNA polymerase. In addition, this compound shows significant inhibition of DNA synthesis in isolated nuclei obtained from adenovirus-infected or uninfected cells and is a potent inhibitor of HeLa cell DNA polymerase alpha. PMID:172658

  16. Genetic characterization and geographic distribution of rabies virus isolates in Brazil: identification of two reservoirs, dogs and vampire bats.

    PubMed

    Ito, M; Arai, Y T; Itou, T; Sakai, T; Ito, F H; Takasaki, T; Kurane, I

    2001-06-01

    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 homology of the 203 nucleotides was at least 83.7% among isolates. The main reservoirs were estimated based on the homology of nucleotide sequences. Brazilian rabies virus isolates were clustered into two reservoir groups: dogs and vampire bats. All the dog-related rabies virus isolates showed nucleotide homology greater than 99.0%. Vampire bat-related rabies virus isolates showed nucleotide homology greater than 96.6% and could be further divided into subgroups corresponding to areas where viruses were isolated. These data suggest that circulating rabies variants belong to at least two different genotype clusters in Brazil and that these two clusters are maintained independently among vampire bats and dogs. PMID:11384221

  17. Phylogenetic and Pathotypical Analysis of Two Virulent Newcastle Disease Viruses Isolated from Domestic Ducks in China

    PubMed Central

    Zhang, Shouping; Wang, Xiaoting; Zhao, Changguang; Liu, Dehua; Hu, Yanxin; Zhao, Jixun; Zhang, Guozhong

    2011-01-01

    Two velogenic Newcastle disease viruses (NDV) obtained from outbreaks in domestic ducks in China were characterized in this study. Phylogenetic analysis revealed that both strains clustered with the class II viruses, with one phylogenetically close to the genotype VII NDVs and the other closer to genotype IX. The deduced amino acid sequence of the cleavage site of the fusion (F) protein confirmed that both isolates contained the virulent motif 112RRQK/RRF117 at the cleavage site. The two NDVs had severe pathogenicity in fully susceptible chickens, resulting in 100% mortality. One of the isolates also demonstrated some pathogenicity in domestic ducks. The present study suggests that more than one genotype of NDV circulates in domestic ducks in China and viral transmission may occur among chickens and domestic ducks. PMID:21949828

  18. Isolation of Bluetongue Virus 24 from India - An Exotic Serotype to Australasia.

    PubMed

    Krishnajyothi, Y; Maan, S; Kandimalla, K; Maan, N S; Tutika, R B; Reddy, Y V; Kumar, A; Mrunalini, N; Reddy, G H; Putty, K; Ahmed, S M; Reddy, Y N; Hemadri, D; Singh, K P; Mertens, P P C; Hegde, N R; Rao, P P

    2016-08-01

    Bluetongue (BT) is a viral disease of ruminants and is caused by different serotypes of bluetongue virus (BTV), which is transmitted by several species of Culicoides midges. The disease is endemic in tropical areas, and incursions have been observed in some of the temperate areas. Twenty-seven recognized serotypes of BTV have been reported so far. Some serotype viruses have been shown to circulate in certain geographical areas. BTV-24 has been reported from Africa, the Mediterranean and the Americas, whereas it is exotic to Australasia. Here, we report isolation of BTV-24 from India and show that it has high sequence homology in genome segment 2 with other Western isolates of BTV-24. Entry of this serotype into Australasian region is a cause of concern. PMID:27241307

  19. Genomic, pathogenic, and antigenic comparisons of Getah virus strains isolated in 1978 and 2014 in Japan.

    PubMed

    Nemoto, Manabu; Bannai, Hiroshi; Tsujimura, Koji; Yamanaka, Takashi; Kondo, Takashi

    2016-06-01

    A Getah virus strain isolated during an outbreak in racehorses in Japan in 2014 (14-I-605) was compared with the vaccine strain isolated in 1978 (MI-110). A comparison of the genome sequences of these strains revealed seven amino acid substitutions in non-structural protein 3, and one or two substitutions in each of other non-structural proteins. In contrast, the structural proteins were highly conserved (99.8-99.9 % amino acid sequence identity). Horse antisera raised against the MI-110 strain showed similar virus-neutralization titers against both MI-110 and 14-I-605 strains (512 and 256, respectively). Therefore, antigenic mutation was probably not a direct cause of the outbreak that occurred in 2014. PMID:27016932

  20. Genome Sequences of Three Apple chlorotic leaf spot virus Isolates from Hawthorns in China.

    PubMed

    Guo, Wei; Zheng, Wenyan; Wang, Mei; Li, Xiaohong; Ma, Yue; Dai, Hongyan

    2016-01-01

    The genome sequences of Apple chlorotic leaf spot virus (ACLSV) isolates from three accessions of hawthorns (Crataegus pinnatifida) grown at Shenyang Agricultural University were determined using Illumina RNA-seq. To confirm the assembly data from the de novo sequencing, two ACLSV genomic sequences (SY01 and SY02) were sequenced using the Sanger method. The SY01 and SY02 sequences obtained with the Sanger method showed 99.5% and 99.7% nucleotide identity with the transcriptome data, respectively. The genome sequences of the hawthorn isolates SY01, SY02 and SY03 (GenBank accession nos. KM207212, KU870524 and KU870525, respectively) consisted of 7,543, 7,561 and 7,545 nucleotides, respectively, excluding poly-adenylated tails. Sequence analysis revealed that these hawthorn isolates shared an overall nucleotide identity of 82.8-92.1% and showed the highest identity of 90.3% for isolate YH (GenBank accession no. KC935955) from pear and the lowest identity of 67.7% for isolate TaTao5 (GenBank accession no. EU223295) from peach. Hawthorn isolate sequences were similar to those of 'B6 type' ACLSV. The relationship between ACLSV isolates largely depends upon the host species. This represents the first comparative study of the genome sequences of ACLSV isolates from hawthorns. PMID:27519059

  1. Genome Sequences of Three Apple chlorotic leaf spot virus Isolates from Hawthorns in China

    PubMed Central

    Zheng, Wenyan; Wang, Mei; Li, Xiaohong; Ma, Yue

    2016-01-01

    The genome sequences of Apple chlorotic leaf spot virus (ACLSV) isolates from three accessions of hawthorns (Crataegus pinnatifida) grown at Shenyang Agricultural University were determined using Illumina RNA-seq. To confirm the assembly data from the de novo sequencing, two ACLSV genomic sequences (SY01 and SY02) were sequenced using the Sanger method. The SY01 and SY02 sequences obtained with the Sanger method showed 99.5% and 99.7% nucleotide identity with the transcriptome data, respectively. The genome sequences of the hawthorn isolates SY01, SY02 and SY03 (GenBank accession nos. KM207212, KU870524 and KU870525, respectively) consisted of 7,543, 7,561 and 7,545 nucleotides, respectively, excluding poly-adenylated tails. Sequence analysis revealed that these hawthorn isolates shared an overall nucleotide identity of 82.8–92.1% and showed the highest identity of 90.3% for isolate YH (GenBank accession no. KC935955) from pear and the lowest identity of 67.7% for isolate TaTao5 (GenBank accession no. EU223295) from peach. Hawthorn isolate sequences were similar to those of ‘B6 type’ ACLSV. The relationship between ACLSV isolates largely depends upon the host species. This represents the first comparative study of the genome sequences of ACLSV isolates from hawthorns. PMID:27519059

  2. Antigenic and genetic diversity of highly pathogenic avian influenza A (H5N1) viruses isolated in Egypt.

    PubMed

    Balish, Amanda L; Davis, C Todd; Saad, Magdi D; El-Sayed, Nasr; Esmat, Hala; Tjaden, Jeffrey A; Earhart, Kenneth C; Ahmed, Lu'ay E; Abd El-Halem, Mohamed; Ali, Abdel Hakem M; Nassif, Samir A; El-Ebiary, Elham A; Taha, M; Aly, Mona M; Arafa, Abdelstattar; O'Neill, Eduardo; Xiyan, Xu; Cox, Nancy J; Donis, Ruben O; Klimov, Alexander I

    2010-03-01

    Highly pathogenic avian influenza A virus (H5N1) has diverged antigenically and genetically since its initial detection in Asia in 1997. Viruses belonging to clade 2.2 in particular have been reported in numerous countries with the majority occurring in Egypt. Previous reports identified antigenic similarities between viruses belonging to clade 2.2. However, poultry and human viruses isolated in northern Egypt during 2007 and 2008 were found to be antigenically distinct from other clade 2.2 viruses from this country. Genetic analysis of the hemagglutinin revealed a high degree of nucleotide and amino acid divergence. The antigenic changes in Egyptian viruses isolated during 2007-08 necessitated that two of these strains be considered as potential H5N1 pre-pandemic vaccine candidates. PMID:20521654

  3. Isolation and molecular characterization of Fikirini rhabdovirus, a novel virus from a Kenyan bat.

    PubMed

    Kading, Rebekah C; Gilbert, Amy T; Mossel, Eric C; Crabtree, Mary B; Kuzmin, Ivan V; Niezgoda, Michael; Agwanda, Bernard; Markotter, Wanda; Weil, M Ryan; Montgomery, Joel M; Rupprecht, Charles E; Miller, Barry R

    2013-11-01

    Zoonotic and vector-borne pathogens have comprised a significant component of emerging human infections in recent decades, and bats are increasingly recognized as reservoirs for many of these disease agents. To identify novel pathogens associated with bats, we screened tissues of bats collected in Kenya. Virus isolates were identified by next generation sequencing of viral nucleic acid preparations from the infected cell culture supernatant and characterized. Here we report the identification of Fikirini rhabdovirus, a novel rhabdovirus isolated from a bat, Hipposideros vittatus, captured along the Kenyan coast. PMID:23939976

  4. Comparative Sequence Analyses of La Crosse Virus Strain Isolated from Patient with Fatal Encephalitis, Tennessee, USA

    PubMed Central

    Fryxell, Rebecca Trout; Freyman, Kimberly; Ulloa, Armando; Velez, Jason O.; Paulsen, Dave; Lanciotti, Robert S.; Moncayo, Abelardo

    2015-01-01

    We characterized a La Crosse virus (LACV) isolate from the brain of a child who died of encephalitis-associated complications in eastern Tennessee, USA, during summer 2012. We compared the isolate with LACV sequences from mosquitoes collected near the child’s home just after his postmortem diagnosis. In addition, we conducted phylogenetic analyses of these and other sequences derived from LACV strains representing varied temporal, geographic, and ecologic origins. Consistent with historical findings, results of these analyses indicate that a limited range of LACV lineage I genotypes is associated with severe clinical outcomes. PMID:25898269

  5. Characterization of a Sorghum mosaic virus (SrMV) isolate in China.

    PubMed

    Zhang, Yu Liang; Pennerman, Kayla K; Wang, Hongxing; Yin, Guohua

    2016-03-01

    Sorghum mosaic virus (SrMV), a causal agent of the destructive sugarcane mosaic disease, has a global presence. An isolate of SrMV infecting a commercially-grown sugarcane plant was recovered from the Hainan province of China. The virions were visualized by an electron microscope, and the coat proteins (CPs) were sequenced by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and tandem mass spectrometry. Discrepancies between the CP predicted and actual amino acid sequences were noted, which confounded the phylogenetic assignment of the isolate. The apparent variations may have physiological effects on the pathogenicity and virulence of SrMV. PMID:26981005

  6. 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

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

    2014-01-01

    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

  7. Isolation and Phylogenetic Analysis of Mucambo Virus (Venezuelan Equine Encephalitis Complex Subtype IIIA) in Trinidad

    PubMed Central

    Auguste, Albert J.; Volk, Sara M.; Arrigo, Nicole C.; Martinez, Raymond; Ramkissoon, Vernie; Adams, A. Paige; Thompson, Nadin N.; Adesiyun, Abiodun A.; Chadee, Dave D.; Foster, Jerome E.; Travassos Da Rosa, Amelia P.A.; Tesh, Robert B.; Weaver, Scott C.; Carrington, Christine V. F.

    2009-01-01

    In the 1950s and 1960s, alphaviruses in the Venezuelan equine encephalitis (VEE) antigenic complex were the most frequently isolated arboviruses in Trinidad. Since then, there has been very little research performed with these viruses. Herein, we report on the isolation, sequencing, and phylogenetic analyses of Mucambo virus (MUCV; VEE complex subtype IIIA), including 6 recently isolated from Culex (Melanoconion) portesi mosquitoes and 11 previously isolated in Trinidad and Brazil. Results show that nucleotide and amino acid identities across the complete structural polyprotein for the MUCV isolates were 96.6 – 100% and 98.7 – 100%, respectively, and the phylogenetic tree inferred for MUCV was highly geographically- and temporally- structured. Bayesian analyses suggest the sampled MUCV lineages have a recent common ancestry of approximately 198 years (with a 95% highest posterior density (HPD) interval of 63 – 448 years) prior to 2007, and an overall rate of evolution of 1.28 × 10−4 substitutions/site/yr. PMID:19631956

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

    PubMed

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

    2014-06-01

    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. PMID:25289003

  9. Nucleotide sequence of both genomic RNAs of a North American tobacco rattle virus isolate.

    PubMed

    Sudarshana, M R; Berger, P H

    1998-01-01

    The complete sequence of a North American tobacco rattle virus (TRV) isolate, 'Oregon yellow' (ORY), was determined from cDNA and RT-PCR clones derived from the two genomic RNAs of this isolate. The RNA-1 is 6790 bases and RNA-2 is 3261 bases. The sequence of TRV-ORY RNA-1 was similar to RNA-1 to TRV isolate SYM, and differs in 48 nucleotides. TRV-ORY RNA-1 was one base shorter than--SYM, and had 47 base substitutions resulting in 12 amino acid substitutions of which 4 were conservative. The RNA-2 of TRV-ORY was distinct from RNA-2 of other characterized TRV isolates and contained three open reading frames (ORFs) that could potentially code for proteins of MW 22.4 kDa, 37.6 kDa and 17.9 kDa. Based on the homology of the predicted amino acid sequence with those of other tobraviruses. ORF1 of RNA-2 encodes the coat protein (CP). The protein sequence of ORF2 had regions of limited similarity with those of ORF2 of two other TRV isolates and pea early browning tobravirus. The ORF3 was unique to TRV-ORY. Phylogenetic analysis of tobravirus CPs indicated that TRV-ORY was most closely related to pepper ringspot tobravirus and TRV-TCM. The relationship of tobravirus CPs to other rod-shaped tubular plant viruses is also discussed. PMID:9739332

  10. Screening, isolation and optimization of anti–white spot syndrome virus drug derived from terrestrial plants

    PubMed Central

    Ghosh, Upasana; Chakraborty, Somnath; Balasubramanian, Thangavel; Das, Punyabrata

    2014-01-01

    Objective To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various terrestrial plants and to evaluate the efficacy of the same in host–pathogen interaction model. Methods Thirty 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. The best anti–WSSV plant isolate, TP22C was isolated and further analyzed. 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 Seven plant isolates exhibited significant survivability in host. The drug TP22C thus formulated showed 86% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of TP22C required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 750 mg/kg body weight/day survived at the rate of 86%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection. Conclusions The drug TP22C derived from Momordica charantia is a potent anti-white spot syndrome virus drug. PMID:25183066

  11. The distribution of African swine fever virus isolated from Ornithodoros moubata in Zambia.

    PubMed Central

    Wilkinson, P. J.; Pegram, R. G.; Perry, B. D.; Lemche, J.; Schels, H. F.

    1988-01-01

    African swine fever (ASF) has been reported in the Eastern Province of Zambia since 1912 and is now considered to be enzootic there. A survey of the distribution of ASF virus in Zambia was carried out by virus isolation from Ornithodoros moubata ticks collected from animal burrows in National Parks and Game Management Areas in northern, eastern, central and southern Zambia. ASF virus was isolated from ticks in all areas examined. The prevalence of infection in O. moubata was between 0.4% in South Luangwa National Park and 5.1% in Livingstone Game Park and mean infectious virus titres ranged from 10(3.4) HAD50/tick in Kakumbe Game Management Area to 10(5.9) HAD50/tick in Chunga and Nalusanga Game Management Areas. The prevalence of infection in adult ticks was between 4.7% and 5.3% in all areas examined except Sumbu National Park and Livingstone Game Park, where the prevalence was 15.1% and 13.2% respectively in adult ticks. The ratio of infected females to males for all the infected adult ticks in all areas of Zambia was 3.2:1. PMID:3215286

  12. Isolation and pathogenic analysis of virulent Marek's disease virus field strain in China.

    PubMed

    Cui, Ning; Su, Shuai; Sun, Peng; Zhang, Yankun; Han, Ni; Cui, Zhizhong

    2016-07-01

    Marek's disease (MD) has become increasingly common in China, resulting in considerable economic loss. The etiological agent is unclear. In this study, we isolated a field MD virus (MDV) strain, designated SX1301, from CVI988/Rispens-vaccinated chickens with tumors. Co-infection of avian leukosis virus, reticuloendotheliosis virus, and chicken infectious anemia virus was excluded by polymerase chain reaction, enzyme-linked immunosorbant assay, DNA blotting hybridization, and indirect immunofluorescence assay. As with most strains isolated in China, SX1301 had the same amino acid mutation of meq protein at positions 77(E), 80(Y), and 115(A) Animal experimental results showed development of lethal MD in 57% and MD tumor in 23% of the specific pathogen-free chickens inoculated with SX1301, with tumors mainly distributed in spleen, liver, and kidney. CVI988/Rispens protected 83% of chickens upon challenge with SX1301, with a mortality rate and tumor incidence of 10% and 7%, respectively. These results implicated SX1301 as a virulent MDV strain, with commercial MDV vaccine CVI988/Rispens unable to confer adequate protection against SX1301. There have been no reports of very virulent (vv) plus MDV in China, but frequently occurring virulent MDV may account for the repeated outbreaks of MD. Vaccines with greater efficacy are needed to protect against MDV. PMID:26976907

  13. Isolation and characterization of tick-borne encephalitis virus from Ixodes persulcatus in Mongolia in 2012.

    PubMed

    Muto, Memi; Bazartseren, Boldbaatar; Tsevel, Bazartseren; Dashzevge, Erdenechimeg; Yoshii, Kentaro; Kariwa, Hiroaki

    2015-07-01

    Tick-borne encephalitis virus (TBEV) is a zoonotic virus belonging to the genus Flavivirus, in the family Flaviviridae. The virus, which is endemic in Europe and northern parts of Asia, causes severe encephalitis. Tick-borne encephalitis (TBE) has been reported in Mongolia since the 1980s, but details about the biological characteristics of the endemic virus are lacking. In this study, 680 ticks (Ixodes persulcatus) were collected in Selenge aimag, northern Mongolia, in 2012. Nine Mongolian TBEV strains were isolated from tick homogenates. A sequence analysis of the envelope protein gene revealed that all isolates belonged to the Siberian subtype of TBEV. Two strains showed similar growth properties in cultured cells, but their virulence in mice differed. Whole genome sequencing revealed only thirteen amino acid differences between these Mongolian TBEV strains. Our results suggest that these naturally occurring amino acid mutations affected the pathogenicity of Mongolian TBEV. Our results may be an important platform for monitoring TBEV to evaluate the epidemiological risk in TBE endemic areas of Mongolia. PMID:26025267

  14. Isolation of Infectious Bursal Disease Virus Using Indigenous Chicken Embryos in Kenya

    PubMed Central

    Mutinda, W. U.; Njagi, L. W.; Nyaga, P. N.; Bebora, L. C.; Mbuthia, P. G.; Kemboi, D.; Githinji, J. W. K.; Muriuki, A.

    2015-01-01

    Infectious bursal disease virus (IBDV) isolates were recovered from outbreaks to initiate activities towards developing a local vaccine strain. Use of indigenous chicken embryos was exploited to determine their potential, promote utilization of local resources for research, and enhance household economic activities. Bursa of Fabricius (BFs) samples from outbreaks shown to be IBDV positive was homogenized and inoculated in 4-week-old specific pathogen-free (SPF) IBDV seronegative white leghorn chicks. The harvested virus was inoculated into 11-day-old indigenous chicken embryos that were IBDV seronegative and passaged serially three times after which they were inoculated into 4-week-old indigenous chicks to test for presence and virulence of propagated virus. Out of 153 BFs collected from outbreaks, 43.8% (67/153) were positive for IBDV antigen and 65.7% (44/67) caused disease in SPF chicks. The embryo mean mortalities were 88% on primary inoculation, 94% in 1st passage, 91% in 2nd passage, and 67% in 3rd passage. After the third passage in embryos all the 44 isolates were virulent in 4-week-old indigenous chicks. The results show that indigenous chicken embryos support growth of IBDV and can be used to propagate the virus as an alternative viral propagating tool for respective vaccine preparation. PMID:27347520

  15. Genetic Diversity of Pneumocystis carinii Isolated from Human Immunodeficiency Virus-Positive Patients in Turin, Italy

    PubMed Central

    Volpe, Gisella; Sbaiz, Luca; Avanzini, Claudio; Caramello, Pietro; Savoia, Dianella

    2001-01-01

    By DNA sequence analysis we identified two new strain types and five novel sporadic variations among 25 isolates of Pneumocystis carinii f. sp. hominis obtained from 19 human immunodeficiency virus-positive patients. Of these, 13 were infected with a single strain and 6 were coinfected. Fifteen different combination types were identified among the 18 strains for which complete molecular typing was accomplished. PMID:11474032

  16. Genetic Structure and Molecular Variability of Cucumber mosaic virus Isolates in the United States

    PubMed Central

    Nouri, Shahideh; Arevalo, Rafael; Falk, Bryce W.; Groves, Russell L.

    2014-01-01

    Cucumber mosaic virus (CMV) has a worldwide distribution and the widest host range of any known plant virus. From 2000 to 2012, epidemics of CMV severely affected the production of snap bean (Phaseulos vulgaris L.) in the Midwest and Northeastern United States. Virus diversity leading to emergence of new strains is often considered a significant factor in virus epidemics. In addition to epidemics, new disease phenotypes arising from genetic exchanges or mutation can compromise effectiveness of plant disease management strategies. Here, we captured a snapshot of genetic variation of 32 CMV isolates collected from different regions of the U.S including new field as well as historic isolates. Nucleotide diversity (π) was low for U.S. CMV isolates. Sequence and phylogenetic analyses revealed that CMV subgroup I is predominant in the US and further showed that the CMV population is a mixture of subgroups IA and IB. Furthermore, phylogenetic analysis suggests likely reassortment between subgroups IA and IB within five CMV isolates. Based on phylogenetic and computational analysis, recombination between subgroups I and II as well as IA and IB in RNA 3 was detected. This is the first report of recombination between CMV subgroups I and II. Neutrality tests illustrated that negative selection was the major force operating upon the CMV genome, although some positively selected sites were detected for all encoded proteins. Together, these data suggest that different regions of the CMV genome are under different evolutionary constraints. These results also delineate composition of the CMV population in the US, and further suggest that recombination and reassortment among strain subgroups does occur but at a low frequency, and point towards CMV genomic regions that differ in types of selection pressure. PMID:24801880

  17. Influenza A(H5N8) virus isolation in Russia, 2014.

    PubMed

    Marchenko, Vasiliy Y; Susloparov, Ivan M; Kolosova, Nataliya P; Goncharova, Nataliya I; Shipovalov, Andrey V; Durymanov, Alexander G; Ilyicheva, Tatyana N; Budatsirenova, Lubov V; Ivanova, Valentina K; Ignatyev, Georgy A; Ershova, Svetlana N; Tulyahova, Valeriya S; Mikheev, Valeriy N; Ryzhikov, Alexander B

    2015-11-01

    In this study, we report the isolation of influenza A(H5N8) virus from a Eurasian wigeon (Anas penelope) in Sakha Republic of the Russian Far East. The strain A/wigeon/Sakha/1/2014 (H5N8) has been shown to be pathogenic for mammals. It is similar to the strains that caused outbreaks in wild birds and poultry in Southeast Asia and Europe in 2014. PMID:26306756

  18. Citrus Viruses in Guatemala: Application of Laboratory-Based Assays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In preparation for a citrus certification in Guatemala, there was an urgent need to determine which graft transmissible citrus pathogens were present. Because of the lack of biological indicator plants, Citrus tristeza virus (CTV) and Xylella fastidiosa, causal agent for citrus variegated chlorosis...

  19. Molecular characterization of rabies virus isolates from Mexico: implications for transmission dynamics and human risk.

    PubMed

    De Mattos, C C; De Mattos, C A; Loza-Rubio, E; Aguilar-Setién, A; Orciari, L A; Smith, J S

    1999-10-01

    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 outbreaks and to increase the understanding of rabies epidemiology in the Western Hemisphere. Antigenic and genetic diversity was found in all samples from dogs and dog-related cases, suggesting a long-term endemic situation with multiple, independent cycles of virus transmission. Two isolates from bobcats were antigenically and genetically homologous to the rabies variant circulating in the Arizona gray fox population, indicating a wider distribution of this variant than previously reported. Rabies isolates from skunks were unrelated to any variant analyzed in this study and represent a previously unrecognized cycle of rabies transmission in skunks in Baja California Sur. Two antigenic and genetic variants co-circulating in southern and eastern Mexico were found in viruses obtained from cases epidemiologically related to vampire bats. These results serve as a baseline for the better understanding of the molecular epidemiology of rabies in Mexico. PMID:10548293

  20. First isolation and genotyping of viruses from recent outbreaks of viral haemorrhagic septicaemia (VHS) in Slovenia.

    PubMed

    Toplak, Ivan; Hostnik, Peter; Rihtaric, Danijela; Olesen, Niels Jørgen; Skall, Helle Frank; Jencic, Vlasta

    2010-10-26

    In November and December 2007, the virus causing viral haemorrhagic septicaemia (VHS) was detected in rainbow trout Oncorhynchus mykiss from 2 fish farms in Slovenia. During 2008 and 2009 the infection spread only among rainbow trout farms and 4 new outbreaks were confirmed. High mortality and clinical signs of VHS were observed among the diseased fish. VHSV was confirmed by virus isolation, immunoperoxidase test, reverse transcriptase polymerase chain reaction (RT-PCR) and phylogenetic analysis. Based on 1 complete (1524 nucleotides [nt]) and 9 partial (600 nt) glycoprotein gene nucleotide sequences, 9 VHSV isolates from the 6 VHS outbreaks were genetically closely related (99 to 100% identity), and were classified into the Subgroup I-a of Genotype I, most closely related to the German isolates Dstg21-07, Dstg36-06, and Dstg54-1-07 (99 to 100% identity). Phylogenetic analysis and epidemiological investigations confirmed that the VHS virus had been (re)introduced with imported live fish, and that subsequent outbreaks were linked to the initial infection. Our study shows that direct nucleotide sequencing of RT-PCR products, amplified from the tissue of VHSV-infected fish, represents a reliable tool for fast routine genotyping in diagnostic laboratories. This is the first report of a natural epidemic associated with VHSV infection in Slovenia since the eradication of the disease in 1977. PMID:21166311

  1. Nasal Wipes for Influenza A Virus Detection and Isolation from Swine.

    PubMed

    Nolting, Jacqueline M; Szablewski, Christine M; Edwards, Jody L; Nelson, Sarah W; Bowman, Andrew S

    2015-01-01

    Surveillance for influenza A viruses in swine is critical to human and animal health because influenza A virus rapidly evolves in swine populations and new strains are continually emerging. Swine are able to be infected by diverse lineages of influenza A virus making them important hosts for the emergence and maintenance of novel influenza A virus strains. Sampling pigs in diverse settings such as commercial swine farms, agricultural fairs, and live animal markets is important to provide a comprehensive view of currently circulating IAV strains. The current gold-standard ante-mortem sampling technique (i.e. collection of nasal swabs) is labor intensive because it requires physical restraint of the pigs. Nasal wipes involve rubbing a piece of fabric across the snout of the pig with minimal to no restraint of the animal. The nasal wipe procedure is simple to perform and does not require personnel with professional veterinary or animal handling training. While slightly less sensitive than nasal swabs, virus detection and isolation rates are adequate to make nasal wipes a viable alternative for sampling individual pigs when low stress sampling methods are required. The proceeding protocol outlines the steps needed to collect a viable nasal wipe from an individual pig. PMID:26709840

  2. A cytoplasmic polyhedrosis virus isolated from the pine processionary caterpillar, Thaumetopoea pityocampa.

    PubMed

    Ince, Ikbal Agah; Demir, Ismail; Demirbag, Zihni; Nalcacioglu, Remziye

    2007-04-01

    A cytoplasmic polyhedrosis virus (CPV) was isolated from the larvae of Thaumetopoea pityocampa and shown to cause an infection of midgut cells. This viral infection revealed several important diagnostic symptoms, including discoloration of the posterior midgut, reduced feeding, and extended development time of the larvae. The virus infection is lethal to Thaumetopoea pityocampa, and with the increasing doses kills the larvae within 4-5 days post infection. Electron microscopy studies showed typical cytoplasmic polyhedral inclusion bodies that are icosahedral, and ranged from 2.4 to 5.3 microm in diameter. Electrophoretic analysis of the RNA genome showed that the virus has a genome composed of 10 equimolar RNA segments with the sizes of 3,907, 3,716, 3,628, 3,249, 2,726, 1,914, 1,815, 1,256, 1,058, and 899 bp, respectively. Based on morphology and nucleic acid analysis, this virus was named Thaumetopoea pityocampa cytoplasmic polyhedrosis virus (TpCPV), and belongs to the genus Cypovirus, family Reoviridae. PMID:18051275

  3. Variable Genome Sequences of the Murine Pneumotropic Virus (Polyomaviridae) Regulatory Region Isolated from an Infected Mouse Tissue Viral Suspension

    PubMed Central

    Libbey, Jane E.

    2016-01-01

    The murine pneumotropic virus genome, isolated from an infected murine tissue homogenate, was sequenced to completion. The lungs, liver, spleen, and kidneys were the source of the tissue homogenate in order to mirror the heterogeneity of the virus population in vivo. The regulatory region sequence was found to be highly variable. PMID:27231357

  4. Characaterization of H5N1 highly pathogenic avian influenza viruses isolated from poultry in Pakistan 2006-2008

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Nine avian influenza viruses (AIV), H5N1 subtype, were isolated from dead poultry in the Karachi region of Pakistan from 2006-2008. The intravenous pathogenicity indices and HA protein cleavage sites of all nine viruses were consistent with highly pathogenic AIV. Based on phylogenetic analysis of ...

  5. Phylogenetic and biological characterization of virulent Newcastle disease viruses isolated in wild birds during 2002-2007

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As part of a West Nile virus surveillance program in the Houston Metropolitan Area and in Rhode Island, extracts from brain from 5608 dead birds representing 21 avian orders, were cultured in Vero cells. Sixteen Newcastle disease virus isolates were recovered from birds of the order Columbiformes. ...

  6. Variable Genome Sequences of the Murine Pneumotropic Virus (Polyomaviridae) Regulatory Region Isolated from an Infected Mouse Tissue Viral Suspension.

    PubMed

    Libbey, Jane E; Fujinami, Robert S

    2016-01-01

    The murine pneumotropic virus genome, isolated from an infected murine tissue homogenate, was sequenced to completion. The lungs, liver, spleen, and kidneys were the source of the tissue homogenate in order to mirror the heterogeneity of the virus population in vivo The regulatory region sequence was found to be highly variable. PMID:27231357

  7. Isolation of exotic Newcastle disease virus (ENDV) from field collected flies and experimental ENDV infections of three arthropod species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    During the 2002 Exotic Newcastle Disease (END) outbreak in California arthropods were collected from two quarantined backyard poultry premises after removal of END virus infected birds. The END virus (ENDV) isolated from field collected pools of three fly species was found to have >98% homology by ...

  8. Evaluation and optimization of avian embryos and cell culture methods for efficient isolation and propagation of avian influenza viruses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Surveillance of wild bird populations for avian influenza viruses (AIV) contributes to our understanding of AIV evolution and ecology. Both real-time reverse transcriptase polymerase chain reaction (RRT-PCR) and virus isolation in embryonating chicken eggs (ECE) are standard methods for detecting A...

  9. Newcastle disease viruses causing recent outbreaks worldwide show unexpectedly high genetic similarity with historical virulent isolates from the 1940s

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Virulent strains of Newcastle disease virus (NDV) cause Newcastle disease (ND), a devastating disease of poultry and wild birds. Phylogenetic analyses clearly distinguish historical isolates (obtained prior to 1960) from currently circulating viruses of class II genotypes V, VI, VII, and XII throug...

  10. West Nile virus isolated from Virginia opossum (Didelphis virginiana) in Northwest Missouri 2012

    SciTech Connect

    Bosco-Lauth, Angela; Harmon, Jessica; Lash, R. Ryan; Weiss, Sonja; Langevin, Stanley; Savage, Harry; Marvin S. Godsey, Jr.; Burkhalter, Kristen; Root, J. Jeffrey; Gidlewski, Thomas; Nicholson, William; Brault, Aaron C.; Komar, Nicholas

    2014-12-01

    We describe the isolation of West Nile virus (WNV; Flaviviridae, flavivirus) from blood of a Virginia opossum (Didelphis virginiana) collected in northwestern Missouri, USA in August 2012. Furthermore, sequencing determined that the virus was related to lineage 1a WNV02 strains. We discuss the role of wildlife in WNV disease epidemiology.

  11. Changuinola Virus Serogroup, New Genomes within the Genus Orbivirus (Family Reoviridae) Isolated in the Brazilian Amazon Region

    PubMed Central

    Dilcher, Meik; Weidmann, Manfred; Carvalho, Valéria L.; Casseb, Alexandre R.; Silva, Eliana V. P.; Nunes, Keley N. B.; Chiang, Jannifer O.; Martins, Lívia C.; Vasconcelos, Pedro F. C.

    2013-01-01

    We report here the first complete genome sequence of a Changuinola virus (CGLV) serotype Irituia virus (BE AN 28873) isolated from a wild rodent (Oryzomys goeldi) in the municipality of Ipixuna, State of Pará, northern Brazil. All genome segments showed similarity with those belonging to members of the genus Orbivirus, family Reoviridae. PMID:24285662

  12. Molecular characterization of Korean Pepper mottle virus isolates and its relationship to symptom variations.

    PubMed

    Kim, Yu-Jeong; Jonson, Miranda Gilda; Choi, Hong Soo; Ko, Sug-Ju; Kim, Kook-Hyung

    2009-09-01

    The symptom variations among Korean Pepper mottle virus (PepMoV) isolates infecting pepper, tomato and potato were described and the cause of variations in relation to molecular variability were investigated. In addition, the entire genome of the 13 PepMoV isolates, collected from five provinces (Kyonggi, Chungnam, Gyeongnam, Jeonbuk and Jeonnam) in Korea, were determined and compared including the previously reported Korean-Vb isolate and 2 other PepMoV isolates isolated from America (CA and FL). Our results showed that the nucleotide sequence of all Korean isolates tested were nearly identical (98-99%) and only 94% similar to American isolates. In general, the complete nucleotide sequences and deduced polyprotein sequences indicated low genetic variation among isolates showing 0.1-3% nucleotide changes per site. However, based on ratio between nucleotide diversity values in nonsynonymous and synonymous position (dN/dS ratio) surprisingly, P1 and 6K2 genes showed relatively high nucleotide substitution ratio (0.8 and 1.0 nucleotide, respectively). When the 6K2 amino acid were aligned, there were 15 amino acid substitutions found in PepMoV-infected potato and only 1 amino acid change from two isolates of PepMoV-infected bell pepper. Interestingly, three isolates including isolate numbers 731, 205135 and 205136 that possessed different aa changes at 6K2 region also showed distinct symptom differentiation in indicator hosts and cosegregated in the phylogenetic analysis. These results further proved previous studies that P1 and 6K2 genes with other proteins might have some involvement on host specificity and pathogenicity. PMID:19374928

  13. Phylogenetic analysis and pathogenicity of H3 subtype avian influenza viruses isolated from live poultry markets in China

    PubMed Central

    Cui, Hongrui; Shi, Ying; Ruan, Tao; Li, Xuesong; Teng, Qiaoyang; Chen, Hongjun; Yang, Jianmei; Liu, Qinfang; Li, Zejun

    2016-01-01

    H3 subtype influenza A virus is one of the main subtypes that threats both public and animal health. However, the evolution and pathogenicity of H3 avian influenza virus (AIV) circulating in domestic birds in China remain largely unclear. In this study, seven H3 AIVs (four H3N2 and three H3N8) were isolated from poultry in live poultry market (LPM) in China. Phylogenetic analyses of full genomes showed that all viruses were clustered into Eurasian lineage, except N8 genes of two H3N8 isolates fell into North American lineage. Intriguingly, the N8 gene of one H3N8 and PB2, PB1, NP and NS of two H3N2 isolates have close relationship with those of the highly pathogenic H5N8 viruses circulating in Korea and United States, suggesting that the H3-like AIV may contribute internal genes to the highly pathogenic H5N8 viruses. Phylogenetic tree of HA gene and antigenic cross-reactivity results indicated that two antigenically different H3 viruses are circulating in LPM in China. Most of the H3 viruses replicated in mice lung and nasal turbinate without prior adaptation, and the representative H3 viruses infected chickens without causing clinical signs. The reassortment of H3 subtype influenza viruses warrants continuous surveillance in LPM in China. PMID:27270298

  14. Phylogenetic analysis and pathogenicity of H3 subtype avian influenza viruses isolated from live poultry markets in China.

    PubMed

    Cui, Hongrui; Shi, Ying; Ruan, Tao; Li, Xuesong; Teng, Qiaoyang; Chen, Hongjun; Yang, Jianmei; Liu, Qinfang; Li, Zejun

    2016-01-01

    H3 subtype influenza A virus is one of the main subtypes that threats both public and animal health. However, the evolution and pathogenicity of H3 avian influenza virus (AIV) circulating in domestic birds in China remain largely unclear. In this study, seven H3 AIVs (four H3N2 and three H3N8) were isolated from poultry in live poultry market (LPM) in China. Phylogenetic analyses of full genomes showed that all viruses were clustered into Eurasian lineage, except N8 genes of two H3N8 isolates fell into North American lineage. Intriguingly, the N8 gene of one H3N8 and PB2, PB1, NP and NS of two H3N2 isolates have close relationship with those of the highly pathogenic H5N8 viruses circulating in Korea and United States, suggesting that the H3-like AIV may contribute internal genes to the highly pathogenic H5N8 viruses. Phylogenetic tree of HA gene and antigenic cross-reactivity results indicated that two antigenically different H3 viruses are circulating in LPM in China. Most of the H3 viruses replicated in mice lung and nasal turbinate without prior adaptation, and the representative H3 viruses infected chickens without causing clinical signs. The reassortment of H3 subtype influenza viruses warrants continuous surveillance in LPM in China. PMID:27270298

  15. A highly divergent Encephalomyocarditis virus isolated from nonhuman primates in Singapore

    PubMed Central

    2013-01-01

    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

  16. Genomic Characterization of a Novel Virus of the Family Tymoviridae Isolated from Mosquitoes

    PubMed Central

    Fu, Shihong; Wang, David; Rayner, Simon; Tang, Qing; Liang, Guodong

    2012-01-01

    Background The family Tymoviridae comprises three plant virus genera, including Tymovirus, Marafivirus, and Maculavirus, which are found in most parts of the world and cause severe agricultural losses. We describe a putatively novel member of the family Tymoviridae, which is isolated from mosquitoes (Culex spp.), referred to as CuTLV. Methods and Results The CuTLV was isolated by cell culture, which replicates and causes cytopathic effects in Aedes albopictus C6/36 cells, but not in mammalian BHK-21 or Vero cells. The complete 6471 nucleotide sequence of CuTLV was determined. The genome of CuTLV is predicted to contain three open reading frames (ORFs). The largest ORF1 is 5307 nucleotides (nt) in length and encodes a putative polypeptide of 1769 amino acids (aa), which contains the conserved motifs for the methyltransferase (MTR), Tymovirus endopeptidase (PRO), helicase (HEL), and RNA-dependent RNA polymerase (RdRp) of the replication-associated proteins (RPs) of positive-stranded RNA viruses. In contrast, the ORF1 sequence does not contain the so-called “tymobox” or “marafibox”, the conserved subgenomic RNA promoter present in all tymoviruses or marafiviruses, respectively. ORF2 and ORF3 putatively encode a 248-aa coat protein (CP) and a proline-rich 149-aa polypeptide. The whole genomic nucleotide identity of CuTLV with other members of family Tymoviridae ranged from 46.2% (ChiYMV) to 52.4% (GFkV). Phylogenetic analysis based on the putative RP and CP genes of CuTLV demonstrated that the virus is most closely related to viruses in the genus Maculavirus. Conclusions The CuTLV is a novel virus related to members of the family Tymoviridae, with molecular characters that are distinct from those of tymoviruses, marafiviruses, and other maculaviruses or macula-like viruses. This is the first report of the isolation of a Tymoviridae-like virus from mosquitoes. Further investigations are required to clarify the origin, replication strategy, and the public health

  17. [Isolation, identification and full-length genome sequence analysis of encephalomyocarditis virus from local aardvarks].

    PubMed

    Chang, Hong-Tao; Liu, Hui-Min; He, Xiu-Yuan; Zhao, Jun; Chen, Lu; Wang, Xin-Wei; Yang, Xia; Yao, Hui-Xia; Wang, Chuan-Qing

    2014-07-01

    Encephalomyocarditis virus (EMCV) is a natural epidemic zoonotic pathogen. However, no reports have been published regarding the isolation, identification and full-length genome of EMCV from a local aardvark population. In present study, an EMCV isolate HNXX13 was isolated from aardvarks named Huainan-pig in Henan Province. The systematic identification, full-length genome sequencing and molecular characteristic analysis of the isolate HNXX13 were conducted. The result showed that the isolate was spherical with a diameter of 24-30 nm, neither heat- nor acid-resistant, sensitive to trypsin, insensitive to chloroform, not protected by bivalent cationic, and the specific fluorescence was observed in the cytoplasm of BHK-21 cells infected with the isolate by using indirect fluorescence assay. The full-length genome of EMCV HNXX13 generated a 7 725bp sequence (GenBank: F771002), with 81.0%-99.9% nucleotide identity to reference strains from different animals, and 99.5% with a Chinese reference strain isolated earlier from a commercial pig herd. The phylogenetic tree based on the full-length genome and ORF sequences identified that all EMCV strains were divided into three groups G1, G2 and G3, and strain HNXX13 belonging to the G1 group with other Chinese reference strains. The result also identified that this EMCV infection could cause severe clinical signs in a local aardvark population, and enriches the molecular epidemiological data of EMCV in China. Regional differences exist in EMCV genome and transmission is limited within a certain area. However, the cross-infection and transmission of EMCV between aardvark and mice appears most likely. Mutations have occurred in some amino acids of EMCV strain HNXX13 during the transmission in local aardvark herd and these mutations might make the virus easier to infect the aardvark. PMID:25272589

  18. Genetic diversity of subgenotype 2.1 isolates of classical swine fever virus.

    PubMed

    Gong, Wenjie; Wu, Jianmin; Lu, Zongji; Zhang, Li; Qin, Shaomin; Chen, Fenglian; Peng, Zhicheng; Wang, Qin; Ma, Ling; Bai, Anbin; Guo, Huancheng; Shi, Jishu; Tu, Changchun

    2016-07-01

    As the causative agent of classical swine fever, the economically devastating swine disease worldwide, classical swine fever virus (CSFV) is currently classified into the 11 subgenotypes, of which subgenotype 2.1 is distributed worldwide and showing more genetic diversity than other subgenotypes. Prior to this report, subgenotype 2.1 was divided into three sub-subgenotypes (2.1a-2.1c). To further analyze the genetic diversity of CSFV isolates in China, 39 CSFV isolates collected between 2004 and 2012 in two Chinese provinces Guangxi and Guangdong were sequenced and subjected to phylogenetic analysis together with reference sequences retrieved from GenBank. Phylogenetic analyses based on the 190-nt and/or 1119-nt full length E2 gene fragments showed that current CSFV subgenotype 2.1 virus isolates in the world could be divided into 10 sub-subgenotypes (2.1a-2.1j) and the 39 isolates collected in this study were grouped into 7 of them (2.1a-2.1c and 2.1g-2.1j). Among the 10 sub-subgenotypes, 2.1d-2.1j were newly identified. Sub-subgenotype 2.1d isolates were circulated only in India, however the rest 9 sub-subgenotypes were from China with some of them closely related to isolates from European and neighboring Asian countries. According to the temporal and spatial distribution of CSFV subgenotype 2.1 isolates, the newly classified 10 sub-subgenotypes were further categorized into three groups: dominant sub-subgenotype, minor sub-subgenotype and silent sub-subgenotype, and each sub-subgenotype can be found only in certain geographical areas. Taken together, this study reveals the complex genetic diversity of CSFV subgenotype 2.1 and improves our understanding about the epidemiological trends of CSFV subgenotype 2.1 in the world, particularly in China. PMID:27085291

  19. Complete nucleotide sequence of Alfalfa mosaic virus isolated from alfalfa (Medicago sativa L.) in Argentina.

    PubMed

    Trucco, Verónica; de Breuil, Soledad; Bejerman, Nicolás; Lenardon, Sergio; Giolitti, Fabián

    2014-06-01

    The complete nucleotide sequence of an Alfalfa mosaic virus (AMV) isolate infecting alfalfa (Medicago sativa L.) in Argentina, AMV-Arg, was determined. The virus genome has the typical organization described for AMV, and comprises 3,643, 2,593, and 2,038 nucleotides for RNA1, 2 and 3, respectively. The whole genome sequence and each encoding region were compared with those of other four isolates that have been completely sequenced from China, Italy, Spain and USA. The nucleotide identity percentages ranged from 95.9 to 99.1 % for the three RNAs and from 93.7 to 99 % for the protein 1 (P1), protein 2 (P2), movement protein and coat protein (CP) encoding regions, whereas the amino acid identity percentages of these proteins ranged from 93.4 to 99.5 %, the lowest value corresponding to P2. CP sequences of AMV-Arg were compared with those of other 25 available isolates, and the phylogenetic analysis based on the CP gene was carried out. The highest percentage of nucleotide sequence identity of the CP gene was 98.3 % with a Chinese isolate and 98.6 % at the amino acid level with four isolates, two from Italy, one from Brazil and the remaining one from China. The phylogenetic analysis showed that AMV-Arg is closely related to subgroup I of AMV isolates. To our knowledge, this is the first report of a complete nucleotide sequence of AMV from South America and the first worldwide report of complete nucleotide sequence of AMV isolated from alfalfa as natural host. PMID:24510307

  20. Lassa virus isolates from Mali and the Ivory Coast represent an emerging fifth lineage

    PubMed Central

    Manning, John T.; Forrester, Naomi; Paessler, Slobodan

    2015-01-01

    Previous imported cases of Lassa fever (LF) into the United Kingdom from the Ivory Coast and Mali, as well as the detection of Lassa virus (LASV) among the Mastomys natalensis population within Mali has led to the suggestion that the endemic area for LF is expanding. Initial phylogenetic analyses arrange isolates from Mali and the Ivory Coast separately from the classical lineage IV isolates taken from Sierra Leone, Guinea, and Liberia. The availability of full genome sequences continues to increase, allowing for a more complete phylogenetic comparison of the isolates from Mali and the Ivory Coast to the other existing isolates. In this study, we utilized a Bayesian approach to infer the demographic histories of each LASV isolate for which the full sequence was available. Our results indicate that the isolates from Mali and the Ivory Coast group separately from the isolates of lineage IV, comprising a distinct fifth lineage. The split between lineages IV and V is estimated to have occurred around 200–300 years ago, which coincides with the colonial period of West Africa. PMID:26483768

  1. Complete genome sequence of an avian leukosis virus isolate associated with hemangioma and myeloid leukosis in egg-type and meat-type chickens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new virus isolate was separated from a commercial egg-type flock of chickens in China and was determined as subgroup J avian leukosis virus (ALV-J). ALV-J is known to cause myeloid leukosis. But this new isolate of viruses causes both hemangioma and myeloid leukosis in chickens. Hemangioma is an a...

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

    PubMed Central

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

    1981-01-01

    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

  3. Strain variation, based on the hemagglutinin gene, in Norwegian ISA virus isolates collected from 1987 to 2001: indications of recombination.

    PubMed

    Devold, M; Falk, K; Dale, B; Krossøy, B; Biering, E; Aspehaug, V; Nilsen, F; Nylund, A

    2001-11-01

    Infectious salmon anemia (ISA) is caused by a virus that probably belongs to the Orthomyxoviridae and was first recorded in Norway in 1984. The disease has since spread along the Norwegian coast and has later been found in Canada, Scotland, the Faroe Islands, Chile, and the USA. This study presents sequence variation of the hemagglutinin gene from 37 ISA virus isolates, viz. one isolate from Scotland, one from Canada and 35 from Norway. The hemagglutinin gene contains a highly polymorphic region (HPR), which together with the rest of the gene sequence provides a good tool for studies of epizootics. The gene shows temporal and geographical sequence variation, where certain areas are dominated by distinct groups of isolates. Evidence of transmission of ISA virus isolates within and between regions is given. It is suggested that the hemagglutinin gene from different isolates may recombine. Possible recombination sites are found within the HPR and in the 5'-end flanking region close to the HPR. PMID:11775793

  4. Genetic and biological characterization of Muko virus, a new distinct member of the species Great Island virus (genus Orbivirus, family Reoviridae), isolated from ixodid ticks in Japan.

    PubMed

    Ejiri, Hiroko; Lim, Chang-Kweng; Isawa, Haruhiko; Kuwata, Ryusei; Kobayashi, Daisuke; Yamaguchi, Yukie; Takayama-Ito, Mutsuyo; Kinoshita, Hitomi; Kakiuchi, Satsuki; Horiya, Madoka; Kotaki, Akira; Takasaki, Tomohiko; Maeda, Ken; Hayashi, Toshihiko; Sasaki, Toshinori; Kobayashi, Mutsuo; Saijo, Masayuki; Sawabe, Kyoko

    2015-12-01

    Among the tick-borne orbiviruses (genus Orbivirus, family Reoviridae), 36 serotypes are currently classified within a single virus species, Great Island virus. In this study, we report the first characterization of a tick-borne orbivirus isolated from the tick Ixodes turdus in Japan, which we identified as a new member of the species Great Island virus. The virus isolate, designated Muko virus (MUV), replicated and induced cytopathic effects in BHK-21, Vero E6, and CCL-141 cells and caused high mortality in suckling mice after intracerebral inoculation. Full genome sequence analysis showed that MUV shared the greatest phylogenetic similarity with Tribeč virus in terms of the amino acid sequences of all viral proteins except for outer capsid protein 1 (OC1; VP4 of MUV). Analysis of genome segment 9 in MUV detected an uninterrupted open reading frame that overlaps with VP6 (Hel), which putatively encodes a molecular and functional equivalent of NS4 from Great Island virus. Our study provides new insights into the geographic distribution, genetic diversity, and evolutionary history of the members of the species Great Island virus. PMID:26350980

  5. Complete coding sequence and molecular epidemiological analysis of Sindbis virus isolates from mosquitoes and humans, Finland.

    PubMed

    Sane, Jussi; Kurkela, Satu; Putkuri, Niina; Huhtamo, Eili; Vaheri, Antti; Vapalahti, Olli

    2012-09-01

    Sindbis virus (SINV) is an arthropod-borne alphavirus, which causes rash-arthritis, particularly in Finland. SINV is transmitted by mosquitoes in Finland but thus far no virus has been isolated from mosquitoes. In this study, we report the isolation of the first SINV strain from mosquitoes in Finland and its full-length protein-coding sequence. We furthermore describe the full-length coding sequence of six SINV strains previously isolated from humans in Finland and from a mosquito in Russia. The strain isolated from mosquitoes (Ilomantsi-2005M) was very closely related to all the other Northern European SINV strains. We found 9 aa positions, of which five in the nsP3 protein C terminus, to be distinctive signatures for the Northern European strains that may be associated with vector or host species adaptation. Phylogenetic analyses further indicate that SINV has a local circulation in endemic regions in Northern Europe and no novel strains are frequently being introduced. PMID:22647374

  6. Phylogenetic analysis of some Newcastle disease virus isolates from the Sudan

    PubMed Central

    Elmardi, N.A.; Bakheit, M.A.; Khalafalla, A.I.

    2016-01-01

    A reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify 1412 bp of the fusion protein gene (F gene) of four Newcastle disease virus (NDV) isolates; two velogenic (TY-1/90 and DIK-90) and two lentogenic isolates (Dongla 88/1 and GD.S.1). Following sequencing, nucleotide sequences were annotated and 894 bp were compared phylogenetically with those from strains previously reported in the Sudan and the virus strains published on the GenBank. It could be demonstrated that TY-1/90 and DIK-90 strains belong to the genotype VI of NDV and are in close genetic relationship to sub- genotype VIb. TY-1/90 and DIK-90 strains were observed to be genetically unrelated to the earlier Sudanese isolates of 1970/80s and the late of 2000s suggesting a different origin. The close genetic relationship to the European and African pigeon paramyxovirus type 1 (PPMV-1) suggests a common ancestor. Dongola, GD.S.1 strains were classified into genotype II that comprises non-pathogenic lentogenic NDV strains. The present genetic classification of NDV isolates of the Sudan provides valuable information on genotypes of NDV. Further molecular epidemiological investigations of the recent outbreaks of Newcastle disease in the Sudan are needed in order to improve the efficiency of control strategies and vaccine development. PMID:27419101

  7. Genetic and antigenic characterization of bovine viral diarrhoea virus type 2 isolated from cattle in India.

    PubMed

    Behera, Sthita Pragnya; Mishra, Niranjan; Vilcek, Stefan; Rajukumar, Katherukamem; Nema, Ram Kumar; Prakash, Anil; Kalaiyarasu, S; Dubey, Shiv Chandra

    2011-03-01

    Previous studies have shown that bovine viral diarrhoea virus type 1 (BVDV-1) subtype b is predominantly circulating in Indian cattle. During testing for exotic pestiviruses between 2007 and 2010, BVDV-2 was identified by real time RT-PCR in two of 1446 cattle blood samples originating from thirteen states of India. The genetic analysis of the isolated virus in 5' UTR, N(pro), entire structural genes (C, E(rns), E1 and E2), nonstructural genes NS2-3 besides 3' UTR demonstrated that the nucleotide and amino acid sequences showed highest similarity with BVDV-2. The entire 5' and 3' UTR consisted of 387 and 204 nucleotides, respectively, and an eight nucleotide repeat motif was found twice within the variable part of 3' UTR that may be considered as a characteristic of BVDV-2. The phylogenetic analysis revealed that the cattle isolate and earlier reported goat BVDV-2 isolate fall into separate clades within BVDV-2a subtype. Antigenic typing with monoclonal antibodies verified the cattle isolate also as BVDV-2. In addition, cross-neutralization tests using antisera raised against Indian BVDV strains circulating in ruminants (cattle, sheep, goat and yak) displayed significant antigenic differences only between BVDV-1 and BVDV-2 strains. This is the first identification of BVDV-2 in Indian cattle that may have important implications for immunization strategies and molecular epidemiology of BVD. PMID:21112633

  8. Isolation and molecular and phylogenetic analyses of encephalomyocarditis virus from wild boar in central China.

    PubMed

    Liu, Huimin; He, Xiuyuan; Song, Xiaofeng; Xu, Liang; Zhang, Yun; Zhou, Guoli; Zhu, Wenjiao; Chang, Chen; Yin, Zhian; Shi, Yuhang; Wang, Chuanqing; Chang, Hongtao

    2016-06-01

    Encephalomyocarditis virus (EMCV) can infect many host species and cause acute myocarditis and respiratory failure in piglets, reproductive failure in pregnant sows. In this study, an EMCV strain, designated JZ1202, was isolated from semi-captive wild boars that presented with acute myocarditis and sudden death in central China. It was identified by hemagglutination inhibition (HI) assay, reverse transcription polymerase chain reaction (RT-PCR) and genome sequencing. The subsequent results showed that the virus could produce a specific cytopathic effect on BHK cells and could cause clinical symptoms and pathological changes in mice. Complete genome sequencing and multiple sequence alignment indicated that JZ1202 strain was 81.3%-99.9% identical with other isolates worldwide. Phylogenetic analysis of the whole genome, ORF, VP3/VP1 and 3D genes using neighbor-joining method revealed that JZ1202 isolate was grouped into lineage 1. The results of this study confirmed that an EMCV strain JZ 1202 isolated from wild boar in central China was fatal to mice and provided new epidemiologic data on EMCV in China. PMID:26917364

  9. Porcine reproductive and respiratory syndrome virus field isolates differ in in vitro interferon phenotypes.

    PubMed

    Lee, Sang-Myeong; Schommer, Susan K; Kleiboeker, Steven B

    2004-12-01

    Type I interferons (IFN-alpha and -beta) play an important role in the innate host defense against viral infection by inducing antiviral responses. In addition to direct antiviral activities, type I IFN serves as an important link between the innate and adaptive immune response through multiple mechanisms. Therefore, the outcome of a viral infection can be affected by IFN induction and the IFN sensitivity of a virus. North American porcine reproductive and respiratory syndrome virus (PRRSV) field isolates were studied with regard to IFN-alpha sensitivity and induction in order to understand the role of type I IFN in PRRSV pathogenesis. PRRSV isolates were differentially sensitive to porcine recombinant IFN-alpha (rIFN-alpha) and varied in their ability to induce IFN-alpha in porcine alveolar macrophages (PAM) cultures as measured by a porcine IFN-alpha specific ELISA on cell culture supernatants. Fifty-two plaques were purified from three PRRSV isolates (numbers 3, 7, and 12) and tested for IFN sensitivity and IFN induction. Plaque-derived populations were composed of heterogeneous populations in terms of IFN-inducing capacity and sensitivity to rIFN-alpha. When macrophages infected with isolates 3, 7, or 12 were treated with polycytidylic acid (polyI:C), IFN-alpha production was enhanced. Cells infected with isolate 3 and treated with polyI:C showed the most consistent and strongest enhancement of IFN-alpha production. It was demonstrated that the relatively low concentrations of IFN-alpha produced by isolate 3 contributed to the enhanced IFN-alpha synthesis in response to polyI:C. Isolates 7 and 12 significantly suppressed the enhanced IFN-alpha production by isolate 3 in polyI:C treated cells. To determine if suppression was at the level of IFN-alpha transcription, quantitative RT-PCR was performed for IFN-alpha mRNA and compared to GAPDH and cyclophilin mRNA quantification. However, the relative number of IFN-alpha transcript copies did not correlate with IFN

  10. Anuran susceptibilities to ranaviruses: role of species identity, exposure route, and a novel virus isolate.

    PubMed

    Hoverman, Jason T; Gray, Matthew J; Miller, Debra L

    2010-03-01

    Ranaviruses are responsible for widespread amphibian die-offs, particularly with larval anurans. To understand the factors that may be contributing to the emergence of ranaviruses, we conducted 3 experiments that exposed 3 species of larval anurans to either endemic frog virus 3 (FV3) or an FV3-like isolate from a ranaculture facility. Our goals were to (1) determine the susceptibility of each species to each virus, (2) determine whether direct ingestion of virions or exposure to virions in a water bath were similarly lethal routes of transmission, and (3) quantify the effects of exposure duration on disease outcomes. We conducted our research in a controlled aquatic laboratory using a factorial combination of virus isolates, transmission routes, and exposure durations. While ranaviruses can affect many species, we found that larval anurans differ greatly in susceptibility to ranaviruses. Average mortality rates of Cope's gray tree frogs (66%) and pickerel frogs (68%) were similar but 3-fold higher than for eastern narrow-mouthed toads. Direct ingestion of the viruses increased mean infection and mortality rates by 30% and caused death about 2 times faster compared to water bath exposure. However, exposure duration did not impact mean infection or mortality rates. We also found that the ranaculture isolate increased mortality by > 34% compared to FV3. Our results suggest that ranaviruses can rapidly infect and cause disease in multiple amphibian species. Given the risk associated with introducing novel ranaviruses from ranaculture facilities, we recommend that all nations adopt the protocol set forth by the World Organization for Animal Health for testing and certifying that amphibians that are commercially shipped are negative for ranavirus infection. PMID:20402227

  11. Genetic diversity of swine influenza viruses isolated from pigs during 2000 to 2005 in Thailand

    PubMed Central

    Takemae, Nobuhiro; Parchariyanon, Sujira; Damrongwatanapokin, Sudarat; Uchida, Yuko; Ruttanapumma, Ruttapong; Watanabe, Chiaki; Yamaguchi, Shigeo; Saito, Takehiko

    2008-01-01

    Background  Recent studies have revealed the existence of genetic diversity in swine influenza viruses (SIVs) in the world. In Thailand, there has been a little information on the molecular characteristics of the SIVs since the first isolation of viruses of H1N1 and H3N2 subtypes in the late 1970s. Our previous study demonstrated that Thai H1N1 SIVs possessed the classical swine H1 and avian‐like swine N1 genes (Takemae et al., Proceedings of the Options for the Control of Influenza VI.2007;350–353). Objectives  In the present study, we genetically characterized 12 SIVs including those of H1N1, H1N2 and H3N2 subtypes isolated between 2000 and 2005. Methods  We determined the entire nucleotide sequences of the eight gene segments of those isolates. Results  Phylogenetic analysis revealed the existence of nine distinct genotypes amongst the Thai SIVs. These genotypes arose from multiple introductions of classical swine, avian‐like swine and human viruses. The existence of two distinct sublineages within classical swine H1 and NS, avian‐like swine PA and M and human H3 and N2 genes of the Thai SIVs suggested that introduction of viruses of classical swine, avian‐like swine and human origins occurred twice respectively into the Thai pig population. The predominance of avian‐like swine genes amongst the Thai SIVs was evident. In particular, three polymerase (PB1, PB2 and PA) and matrix genes of avian‐like swine origin were retained in all the Thai SIVs examined. Conclusions  These observations may suggest that genes of avian‐like swine lineages have some advantages to be maintained in pigs as seen in the SIVs established through multiple introductions in other regions. PMID:19453423

  12. Isolation of a novel serotype strain of infectious bronchitis virus ZZ2004 from ducks in China.

    PubMed

    Yao, Sixin; Ou, Changbo; Liu, Xingyou; Wang, Xianwen; Yao, Zonghui; Liu, Jinjing

    2016-10-01

    In chickens, the infectious bronchitis virus (IBV) often causes respiratory distress, a decrease in egg production, poor egg quality, and occasional nephritis. However, ZZ2004, a Chinese isolate of IBV, was obtained from ducks with clinical growth suppression and mild respiratory symptoms that had been reared with chickens in the central region of China. Virus isolation, virus neutralization testing, and RT-PCR were employed to identify the causative pathogen, while sequence alignment was used to analyze gene variations of the S1 subunit and M genes. The results showed that the ducks were infected with IBV due to the emergence of a dwarfing phenotype and the death of embryos between 48 and 144 h post-inoculation. RT-PCR also confirmed the presence of the expected fragment sizes of the S1 subunit and M genes by RT-PCR. Meanwhile, the results of the virus neutralization test indicated that the strains of JX/99/01, GD, SAIBK, LDT3 showed cross-reactivity with the ZZ2004 isolate, and hardly any cross-neutralization of IBV ZZ2004 was observed with the strains of M41, H120, Gray, Holte, or Aust-T. Phylogenetic analysis suggested that there were large differences between ZZ2004 and other IBV reference strains on the S1 subunit. Meanwhile, homologies in the nucleotide and amino acid sequences of the M gene of IBV ZZ2004 were 86.9-92.0 % and 91.1-93.9 %, respectively, compared with 35 other IBV reference strains derived from different regions. This result revealed that there were conspicuous variations among the selected strains. Furthermore, the results showed that the prevalent strains of IBV in ducks had no antigen homology with the vaccine strains widely used in China except the LDT3-strain, making it urgent to explore and develop new IBV vaccines. PMID:27164844

  13. Antigenic analysis of SAT 2 serotype foot-and-mouth disease virus isolates from Zimbabwe using monoclonal antibodies.

    PubMed Central

    Davidson, F. L.; Crowther, J. R.; Nqindi, J.; Knowles, N. J.; Thevasagayam, S. J.; Van Vuuren, C. J.

    1995-01-01

    This paper compares strains of foot-and-mouth disease (FMD) serotype SAT (South African Territories) 2 viruses isolated from Zimbabwe and other African countries using monoclonal antibodies (MAb). A sandwich-ELISA was used to examine the relative binding of anti-SAT 2 MAb to the various viruses. The MAb-binding profiles of viruses isolated from field samples were compared using hierarchical cluster analysis. Viruses were obtained from game animals, mainly African buffalo (Syncerus caffer) which is the natural host and reservoir for SAT serotypes in Africa, and from cattle showing clinical signs of FMD, as well as from animals suspected of carrying the virus subclinically. Some isolates have been adapted for use as vaccine strains. The results showed that most of the Zimbabwe isolates collected between 1989 and 1992 were an antigenically closely-related group. Although differences were observed between Zimbabwe isolates collected between 1989 and 1992 and those collected in 1987, there was no correlation with the different MAb binding patterns within the 1987 group and the epidemiological information received from the field. Similar profiles were observed for many SAT 2 viruses, including viruses isolated over a 50-year period and from geographically distant areas. This indicates an inherent stability in antigenic profiles of SAT 2 viruses. The MAb panel was capable of assessing antigenic variation, since very different profiles were obtained for some isolates. The work also allowed comparison and characterization of anti-type SAT 2 MAb from different laboratories. The findings are discussed with reference to selection of vaccine strains. PMID:7543860

  14. Rift Valley fever virus (family Bunyaviridae, genus Phlebovirus). Isolations from Diptera collected during an inter-epizootic period in Kenya.

    PubMed Central

    Linthicum, K. J.; Davies, F. G.; Kairo, A.; Bailey, C. L.

    1985-01-01

    A total of 134 876 Diptera collected in Kenya during a 3-year period were tested in 3383 pools for Rift Valley fever (RVF) virus. Nineteen pools of unengorged mosquitoes were found positive for RVF. All isolations were made from specimens collected at or near the naturally or artificially flooded grassland depressions that serve as the developmental sites for the immature stages of many mosquito species. The isolation of virus from adult male and female A. lineatopennis which had been reared from field-collected larvae and pupae suggests that transovarial transmission of the virus occurs in this species. PMID:2862206

  15. Isolation and complete genome sequencing of Mimivirus bombay, a Giant Virus in sewage of Mumbai, India.

    PubMed

    Chatterjee, Anirvan; Ali, Farhan; Bange, Disha; Kondabagil, Kiran

    2016-09-01

    We report the isolation and complete genome sequencing of a new Mimiviridae family member, infecting Acanthamoeba castellanii, from sewage in Mumbai, India. The isolated virus has a particle size of about 435 nm and a 1,182,200-bp genome. A phylogeny based on the DNA polymerase sequence placed the isolate as a new member of the Mimiviridae family lineage A and was named as Mimivirus bombay. Extensive presence of Mimiviridae family members in different environmental niches, with remarkably similar genome size and genetic makeup, point towards an evolutionary advantage that needs to be further investigated. The complete genome sequence of Mimivirus bombay was deposited at GenBank/EMBL/DDBJ under the accession number KU761889. PMID:27330993

  16. Complete nucleotide sequence of a new isolate of passion fruit woodiness virus from Western Australia.

    PubMed

    Fukumoto, Tomohiro; Nakamura, Masayuki; Wylie, Stephen J; Chiaki, Yuya; Iwai, Hisashi

    2013-08-01

    We determined the complete genome sequence of the passion fruit woodiness virus Gld-1 isolate (PWV-Gld-1) from Australia and compared it with that of PWV-MU-2, another Australian isolate of PWV. The genomes shared high sequence identity in both the complete nucleotide sequence and the ORF amino acid sequence. All of the cleavage sites of each protein were identical to those of MU-2, and the sequence identity for the individual proteins ranged from 97.2 % to 100.0 %. However, the 5' untranslated region (5'UTR) of the Gld-1 isolate shared only 46.8 % sequence identity with that of PWV-MU-2 and was 177 nucleotides shorter. Re-sequencing of the 5'UTR of MU-2 revealed that the 5' end of the original sequence includes an artifact generated by deep sequencing. PMID:23508550

  17. Isolation and identification of bat viruses closely related to human, porcine and mink orthoreoviruses.

    PubMed

    Yang, Xing-Lou; Tan, Bing; Wang, Bo; Li, Wen; Wang, Ning; Luo, Chu-Ming; Wang, Mei-Niang; Zhang, Wei; Li, Bei; Peng, Cheng; Ge, Xing-Yi; Zhang, Li-Biao; Shi, Zheng-Li

    2015-12-01

    Bats have been identified as natural reservoirs of many viruses, including reoviruses. Recent studies have demonstrated the interspecies transmission of bat reoviruses to humans. In this study, we report the isolation and molecular characterization of six strains of mammalian orthoreovirus (MRV) from Hipposideros and Myotis spp. These isolates were grouped into MRV serotype 1, 2 or 3 based on the sequences of the S1 gene, which encodes the outer coat protein s1. Importantly, we found that three of six bat MRV strains shared high similarity with MRVs isolated from diseased minks, piglets or humans based on the S1 segment, suggesting that interspecies transmission has occurred between bats and humans or animals. Phylogenetic analyses based on the 10 segments showed that the genomic segments of these bat MRVs had different evolution lineages, suggesting that these bat MRVs may have arisen through reassortment of MRVs of different origins. PMID:26475793

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

    PubMed

    Chacón, Jorge Luis; Ferreira, Antonio J Piantino

    2009-11-12

    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

  19. Isolation and Characterization of Mayaro Virus from a Human in Acre, Brazil

    PubMed Central

    Terzian, Ana Carolina B.; Auguste, Albert J.; Vedovello, Danila; Ferreira, Marcelo U.; da Silva-Nunes, Mônica; Sperança, Márcia A.; Suzuki, Rodrigo B.; Juncansen, Camila; Araújo, João P.; Weaver, Scott C.; Nogueira, Maurício L.

    2015-01-01

    Mayaro virus (MAYV) is widely distributed throughout South America and is the etiologic agent of Mayaro fever, an acute febrile illness often presenting with arthralgic manifestations. The true incidence of MAYV infection is likely grossly underestimated because the symptomatic presentation is very similar to that of dengue fever and other acute febrile tropical diseases. We report the complete genome sequence of a MAYV isolate detected from an Acrelândia patient presenting with fever, chills, and sweating, but with no arthralgia. Results show that this isolate belongs to genotype D and is closely related to Bolivian strains. Our results suggest that the Acre/Mayaro strain is closely related to the progenitor of these Bolivian strains that were isolated between 2002 and 2006. PMID:25510721

  20. Molecular characterisation of the full-length genome of olive latent virus 1 isolated from tomato.

    PubMed

    Hasiów-Jaroszewska, Beata; Borodynko, Natasza; Pospieszny, Henryk

    2011-05-01

    Olive latent virus 1 (OLV-1) is a species of the Necrovirus genus. So far, it has been reported to infect olive, citrus tree and tulip. Here, we determined and analysed the complete genomic sequence of an isolate designated as CM1, which was collected from tomato plant in the Wielkopolska region of Poland and represents the prevalent isolate of OLV-1. The CM1 genome consists of monopartite single-stranded positive-sense RNA genome sized 3,699 nt with five open reading frames (ORFs) and small inter-cistronic regions. ORF1 encodes a polypeptide with a molecular weight of 23 kDa and the read-through (RT) of its amber stop codon results in ORF1 RT that encodes the virus RNA-dependent RNA polymerase. ORF2 and ORF3 encode two peptides, with 8 kDa and 6 kDa, respectively, which appear to be involved in cell-to-cell movement. ORF4 is located in the 3' terminal and encodes a protein with 30 kDa identified as the viral coat protein (CP). The differences in CP region of four OLV-1 isolates whose sequences have been deposited in GenBank were observed. Nucleotide sequence identities of the CP of tomato CM1 isolate with those of olive, citrus and tulip isolates were 91.8%, 89.5% and 92.5%, respectively. In contrast to other OLV-1 isolates, CM1 induced necrotic spots on tomato plants and elicited necrotic local lesions on Nicotiana benthamiana, followed by systemic infection. This is the third complete genomic sequence of OLV-1 reported and the first one from tomato. PMID:21181333