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We studied the genetic variability of three genomic regions (p23, p25 and p27 genes) from 11 field Citrus tristezavirusisolates from the two main citrus growing areas of Argentina, a country where the most efficient vector of the virus, Toxoptera citricida, is present for decades. The pathogenicity of the isolates was determinated by biological indexing, single-strand conformation polymorphism analysis showed that most isolates contained high intra-isolate variability. Divergent sequence variants were detected in some isolates, suggesting re-infections of the field plants. Phylogenetic analysis of the predominant sequence variants of each isolate revealed similar grouping of isolates for genes p25 and p27. The analysis of p23 showed two groups contained the severe isolates. Our results showed a high intra-isolate sequence variability suggesting that re-infections could contribute to the observed variability and that the host can play an important role in the selection of the sequence variants present in these isolates. PMID:17999168
Iglesias, Néstor G; Gago-Zachert, Selma P; Robledo, Germán; Costa, Norma; Plata, María Inés; Vera, Osmar; Grau, Oscar; Semorile, Liliana C
Technical Abstract: Stem pitting isolates of Citrus tristezavirus (CTV) are not thought to be widely distributed in Florida, but are of concern and a test was needed for rapid discrimination of stem pitting from other severe CTV isolates in field trees. Four published techniques were evaluated f...
The nucleotide sequence diversity of the CPm gene from 28 field isolates of Citrus tristezavirus (CTV) was assessed by SSCP and sequence analyses. These isolates showed two major shared haplotypes, which differed in distribution:\\u000a A1 was the major haplotype in 23 isolates from different geographic regions, whereas R1 was found in isolates from a discrete\\u000a region. Phylogenetic reconstruction clustered
Oscar Arturo Oliveros-Garay; Natalhie Martinez-Salazar; Yanneth Torres-Ruiz; Orlando Acosta
Genetic markers amplified by sequence specific primers designed from the partial or complete genome sequences of Citrus tristezavirus (CTV) isolates T3, T30, T36 and VT were used to assess the genetic relatedness of isolates in an international collection of 372 exotic isolates of CTV by comparing...
An assessment was made of the disease potential of a Citrus tristezavirus (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...
Aphis gossypii is the principal vector of Citrus tristezavirus (CTV) in California. Based on host range, a population of A. gosspyii exists as the melon aphid (MA) or the cotton aphid (CA) biotype. Previous reports on CTV transmission used the MA reared on cucurbits. The objective of this stud...
Citrus tristezavirus (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...
Citrus tristezavirus (CTV) is a phloem-limited virus whose natural host range is restricted to citrus and related species. Although the virus has killed millions of trees, almost destroying whole industries, and continually limits production in many citrus growing areas, most isolates are mild or symptomless in most of their host range. There is little understanding of how the virus causes severe disease in some citrus and none in others. Movement and distribution of CTV differs considerably from that of well-studied viruses of herbaceous plants where movement occurs largely through adjacent cells. In contrast, CTV systemically infects plants mainly by long-distance movement with only limited cell-to-cell movement. The virus is transported through sieve elements and occasionally enters an adjacent companion or phloem parenchyma cell where virus replication occurs. In some plants this is followed by cell-to-cell movement into only a small cluster of adjacent cells, while in others there is no cell-to-cell movement. Different proportions of cells adjacent to sieve elements become infected in different plant species. This appears to be related to how well viral gene products interact with specific hosts. CTV has three genes (p33, p18, and p13) that are not necessary for infection of most of its hosts, but are needed in different combinations for infection of certain citrus species. These genes apparently were acquired by the virus to extend its host range. Some specific viral gene products have been implicated in symptom induction. Remarkably, the deletion of these genes from the virus genome can induce large increases in stem pitting (SP) symptoms. The p23 gene, which is a suppressor of RNA silencing and a regulator of viral RNA synthesis, has been shown to be the cause of seedling yellows (SY) symptoms in sour orange. Most isolates of CTV in nature are populations of different strains of CTV. The next frontier of CTV biology is the understanding how the virus variants in those mixtures interact with each other and cause diseases.
Dawson, W. O.; Garnsey, S. M.; Tatineni, S.; Folimonova, S. Y.; Harper, S. J.; Gowda, S.
The sequence of the entire genome of citrus tristezavirus (CTV), Florida isolate T36, was completed. The 19,296-nt CTV genome encodes 12 open reading frames (ORFs) potentially coding for at least 17 protein products. The 5?-proximal ORF 1a starts at nucleotide 108 and encodes a large polyprotein with calculated MW of 349 kDa containing domains characteristic of (from 5? to
A. V. Karasev; V. P. Boyko; S. Gowda; O. V. Nikolaeva; M. E. Hilf; E. V. Koonin; C. L. Niblett; K. Cline; D. J. Gumpf; R. F. Lee; S. M. Garnsey; D. J. Lewandowski; W. O. Dawson
Citrus tristezavirus (CTV), a phloem-limited closterovirus, contains a single-stranded, 20- kb RNA. Numerous isolates of the virus exist which produce a variety of symptoms in various citrus hosts. One of these symptoms is stem pitting (SP). SP does not occur in all citrus hosts but may cause lo...
Citrus tristezavirus (CTV) exists in field isolates as a complex of virusisolates. This complex may contain both mild and severe CTV. Using single and multiple aphid transmissions, subisolates of the various field isolates were separated. Some CTV isolates that tested negative with the monoclon...
Tristeza is an important citrus disease affecting the viability and productivity of citrus worldwide. The causal agent, Citrus tristezavirus (CTV), usually occurs as a mixture of genotypes in nature, with one of the genotypes often dominating the population. CTV has a monopartite, positive-sense RNA genome of approximately 19.3 kb and exhibits over 30% diversity in the 5' half and less than 10% in the 3' half among different genotypes. A Florida CTV isolate, FS627, was selected for this study. Isolate FS627 was analyzed by reverse-transcription polymerase chain reaction (RT-PCR) using primers to three regions: 788-bp region in the 5' (697 to 1,484 nucleotides), open reading frame (ORF)1a, 696 or 718 bp from the overlapping region of the RdRp (ORF1b) and p33 (ORF2) gene, and a 672-bp major coat protein gene (ORF7) in the 3' end of the CTV genome. The presence of T36, T30, and VT genotypes in isolate FS627 was confirmed utilizing the genotype specific overlapping region of RdRp primer pairs for RT-PCR amplification followed by cloning and sequence analysis. Analysis of single-strand conformational polymorphisms and sequences of RT-PCR-amplified products of the above regions were used to determine the presence of genotypes in both the parent and aphid-transmitted (AT) subisolates. Although the parent isolate had T36 as the major genotype, T30 was the major genotype in most of the AT subisolates. Some intermediate genotypes were detected that differed from the parental or AT subisolates. These intermediate genotypes were considered to be recombinants of the T30 and T36 genotypes and also were observed in the second level of AT subisolates generated from the of first-level AT subisolates of CTV-FS627. This work provides advance information on the population dynamics in CTV mixtures and the generation of virus recombinants after aphid transmission. PMID:19821734
Citrus tristezavirus (CTV) causes severe losses in grapefruit production in South Africa and requires mild-strain cross-protection to maintain production. Unfortunately, cross-protection breakdown of the pre-immunizing CTV grapefruit mild source GFMS12 is prevalent in grapefruit in South Africa. The CTV genotype composition of the GFMS12 population inoculated onto different hosts was determined by sequencing part of ORF1a and the p23 gene of multiple clones from each plant. Analysis of the GFMS12 population in Mexican lime and Marsh and Star Ruby grapefruit varieties revealed that at least four genotypes occur in the GFMS12 population and that genotype compositions differed amongst the populations in different host plants. Single-aphid-transmitted sub-isolates derived from the GFMS12 mother population on Mexican lime appeared to contain three populations of a mixture of VT-like and recombinant B165/VT-like genotypes; a mixture of recombinant RB/VT- and B165/VT-like genotypes; and a single recombinant B165/VT-like genotype. This study underlines the importance of determining the genotype composition of a potential CTV pre-immunizing source on a range of inoculated host species before utilization. PMID:22932923
Scott, Katherine Anne; Hlela, Quinsile; Zablocki, Olivier; Read, David; van Vuuren, Stephanus; Pietersen, Gerhard
Citrus tristezavirus (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...
The Nartia strain of citrus tristezavirus (CTV) has been widely used during last fifty years for cross protection against stem pitting CTV in South African grapefruit. We have previously shown the presence of several genotypes of CTV in this isolate by use of a heteroduplex mobility assay of a 400 ...
Citrus tristezavirus(CTV) is a major viral pathogen of citrus, with a natural and experimental host range confined to members of the family Rutaceae, including the economically important genus Citrus. Experimental and agricultural paths of distribution of CTV are transmission by aphids and graft p...
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 tristezavirus (CTV) infected trees to protect the CCPP and re...
Citrus tristezavirus (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.
Davino, Salvatore; Willemsen, Anouk; Panno, Stefano; Davino, Mario; Catara, Antonino; Elena, Santiago F.; Rubio, Luis
Citrus tristezavirus (CTV) causes two citrus diseases that have caused devastating losses in global citrus production. The first disease is quick decline of trees propagated on the sour orange rootstock. The second disease is stem pitting, which severely affects a number of economically important citrus varieties regardless of the rootstock used and results in reduced tree growth and vigor as well as in reduced fruit size and quality. Both diseases continue to invade new areas. While quick decline could be effectively managed by the use of resistant and/or tolerant rootstocks, the only means to protect commercial citrus against endemic stem pitting isolates of CTV has been cross-protection with mild isolates of the virus. In some citrus areas cross-protection has been successful and allowed production of certain citrus cultivars despite the presence of severe stem pitting isolates in those regions. However, many other attempts to find isolates that would provide sustained protection against aggressive isolates of the virus had failed. In general, there has been no understanding why some mild isolates were effective and others failed to protect. We have been working on the mechanism of cross-protection by CTV. Recent considerable progress has significantly advanced our understanding of how cross-protection may work in the citrus/CTV pathosystem. As we demonstrated, only isolates that belong to the same strain of the virus cross protect against each other, while isolates from different strains do not. We believe that the results of our research could now make finding protecting isolates relatively straightforward. This review discusses some of the history of CTV cross-protection along with the recent findings and our "recipe" for selection of protecting isolates. PMID:23577008
The incidence of Citrus tristezavirus (CTV) was found to increase significantly in southern Florida within two years after the establishment of its most efficient vector, Toxoptera citricida (Kirkaldy). The increased incidence of both mild and severe strains was documented, with the incidence of se...
TECHNICAL ABSTRACT: Aphis gossypii is the primary vector of Citrus tristezavirus (CTV) in California and exists as two distinct biotypes: the melon aphid; and the cotton aphid. These biotypes are morphologically indistinguishable but have distinct host ranges. Both aphids have a wide host range...
A localized genetic linkage map was developed of the region surrounding the citrus tristezavirus (CTV) resistance gene (designated Ctv) from Poncirus trifoliate L., a sexually compatible Citrus relative. Bulked segregant analysis (BSA) was used to identify potential resistance-associated RAPD fragment markers in four intergeneric backcross families that were segregating for CTV resistance. Eight RAPD fragments were found that were
F. G. Gmitter Jr; S. Y. Xiao; S. Huang; X. L. Hu; S. M. Garnsey; Z. Deng
Citrus tristezavirus (CTV) is the most economically important viral disease of citrus worldwide. Cultivars with improved CTV tolerance or resistance are needed to manage CTV-induced diseases. The citrus relatives Poncirus trifoliata (L.) Raf., Swinglea glutinosa (Blanco) Merr., and Severinia buxifolia (Poir) Ten. are potential sources of CTV resistance, but their resistance mechanisms are poorly characterized. As a first step
Maria R. Albiach-Marti; Jude W. Grosser; Siddarame Gowda; Munir Mawassi; Tatineni Satyanarayana; Stephen M. Garnsey; William O. Dawson
The coat protein gene of citrus tristezavirus (CTV) has been introduced into Mexican lime (Citrus aurantifolia Swing.) plants by using an improved Agrobacterium-mediated genetic transformation system. Internodal stem segments from greenhouse-grown seedlings were co-cultivated with\\u000a A. tumefaciens strain EHA 105 carrying the binary plasmid pBI 121\\/CTV-CP in a medium rich in auxins that provided the explant cells with the
A. Domínguez; J. Guerri; M. Cambra; L. Navarro; P. Moreno; L. Peńa
Citrus tristezavirus (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...
The large RNA genome of Citrus tristezavirus (CTV; ca. 20 kb) contains 12 open reading frames, with the 3'-terminal one corresponding to a protein of 209 amino acids (p23) that is expressed from an abundant subgenomic RNA. p23, an RNA-binding protein with a putative zinc-finger domain and some basic motifs, is unique to CTV because no homologs have been found in other closteroviruses, including the type species of the genus Beet yellows virus (despite both viruses having many homologous genes). Consequently, p23 might have evolved for the specific interaction of CTV with its citrus hosts. From a functional perspective p23 has been involved in many roles: (i) regulation of the asymmetrical accumulation of CTV RNA strands, (ii) induction of the seedling yellows syndrome in sour orange and grapefruit, (iii) intracellular suppression of RNA silencing, (iv) elicitation of CTV-like symptoms when expressed ectopically as a transgene in several Citrus spp., and (v) enhancement of systemic infection (and virus accumulation) in sour orange and CTV release from the phloem in p23-expressing transgenic sweet and sour orange. Moreover, transformation of Mexican lime with intron-hairpin constructs designed for the co-inactivation of p23 and the two other CTV silencing suppressors results in complete resistance against the homologous virus. From a cellular point of view, recent data indicate that p23 accumulates preferentially in the nucleolus, being the first closterovirus protein with such a subcellular localization, as well as in plasmodesmata. These major accumulation sites most likely determine some of the functional roles of p23. PMID:23653624
The large RNA genome of Citrus tristezavirus (CTV; ca. 20 kb) contains 12 open reading frames, with the 3?-terminal one corresponding to a protein of 209 amino acids (p23) that is expressed from an abundant subgenomic RNA. p23, an RNA-binding protein with a putative zinc-finger domain and some basic motifs, is unique to CTV because no homologs have been found in other closteroviruses, including the type species of the genus Beet yellows virus (despite both viruses having many homologous genes). Consequently, p23 might have evolved for the specific interaction of CTV with its citrus hosts. From a functional perspective p23 has been involved in many roles: (i) regulation of the asymmetrical accumulation of CTV RNA strands, (ii) induction of the seedling yellows syndrome in sour orange and grapefruit, (iii) intracellular suppression of RNA silencing, (iv) elicitation of CTV-like symptoms when expressed ectopically as a transgene in several Citrus spp., and (v) enhancement of systemic infection (and virus accumulation) in sour orange and CTV release from the phloem in p23-expressing transgenic sweet and sour orange. Moreover, transformation of Mexican lime with intron-hairpin constructs designed for the co-inactivation of p23 and the two other CTV silencing suppressors results in complete resistance against the homologous virus. From a cellular point of view, recent data indicate that p23 accumulates preferentially in the nucleolus, being the first closterovirus protein with such a subcellular localization, as well as in plasmodesmata. These major accumulation sites most likely determine some of the functional roles of p23.
Citrus tristezavirus (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 ...
Stem pitting is a common virus-induced disease of perennial woody plants induced by a range of different viruses. The phenotype results from sporadic areas of the stem in which normal xylem and phloem development is prevented during growth of stems. These alterations interfere with carbohydrate transport, resulting in reduced plant growth and yield. Citrus tristezavirus (CTV), a phloem-limited closterovirus, induces economically important stem-pitting diseases of citrus. CTV has three nonconserved genes (p33, p18, and p13) that are not related to genes of other viruses and that are not required for systemic infection of some species of citrus, which allowed us to examine the effect of deletions of these genes on symptom phenotypes. In the most susceptible experimental host, Citrus macrophylla, the full-length virus causes only very mild stem-pitting symptoms. Surprisingly, we found that certain deletion combinations (p33 and p18 and/or p13) induced greatly increased stem-pitting symptoms, while other combinations (p13 or p13 plus p18) resulted in reduced stem pitting. These results suggest that the stem-pitting phenotype, which is one of more economically important disease phenotypes, can result not from a specific sequence or protein but from a balance between the expression of different viral genes. Unexpectedly, using green fluorescent protein-tagged full-length virus and deletion mutants (CTV9?p33 and CTV9?p33?p18?p13), the virus was found at pitted areas in abnormal locations outside the normal ring of phloem. Thus, increased stem pitting was associated not only with a prevention of xylem production but also with a proliferation of cells that supported viral replication, suggesting that at random areas of stems the virus can elicit changes in cellular differentiation and development. PMID:22593155
Enzyme-linked immunosorbent assay (ELISA) proved to be a sensitive detector for citrus tristezavirus (CTV) in orange fruits\\u000a (Citrus sinensis (L.) Osbeck). Samples of five fruits were taken from 350-kg packing house containers and tested by ELISA to predict the infection\\u000a rate of CTV in two infected orange groves. The predicted infection rates, 1% and 11%, were in reasonable agreement
Transgenic plants of grapefruit (Citrus paradisi Macf.) cvs. ‘Duncan’, ‘Flame’, ‘Marsh’, and ‘Ruby Red’ were obtained using Agrobacterium tumefaciens-mediated transformation of seedling epicotyl tissue. Two citrus tristezavirus (CTV)-derived candidate resistance genes:\\u000a ‘392’ (3? region of the p23 ORF plus 3? untranslated region—UTR) and ‘p23 hairpin’ (sense-p23 ORF plus UTR plus antisense-p23\\u000a ORF) were introduced into grapefruit using Agrobacterium strains
G. Ananthakrishnan; V. Orbovi?; G. Pasquali; M. ?alovi?; J. W. Grosser
The long flexuous bipolar virions of Citrus tristezavirus (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...
A multiplex Taqman-based real-time reverse transcription (RT) polymerase chain reaction (PCR) assay was developed to identify potential severe strains of Citrus tristezavirus (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
A quantitative and multiplex real-time RT-PCR assay was developed to detect Citrus tristezavirus (CTV) along with plant mRNA, which serves as an internal control to ascertain RNA extraction quality. The real-time technique was validated against 39 CTV strains from around the world as well as with the aphid vector, Aphis gossypii, given a 48 h acquisition access period on a CTV source plant. The assay was effective for quantitation of the viral template in infected plants and in single aphids. CTV detection was compared from different plant tissues and for different RNA isolation methods from aphids. Less than 1 fg was consistently detected when RNA transcripts were diluted in extracts from healthy plants while RNA copies carried by single aphids were estimated to be between 12,000 and 13,000,000. The assay was more sensitive and less time consuming than ELISA or traditional RT-PCR. The real-time RT-PCR assay developed is a valuable new tool for detection and titer quantitation of CTV. PMID:17888522
Saponari, Maria; Manjunath, Keremane; Yokomi, Raymond K
Citrus tristezavirus (CTV) encodes a singular protein (p23, 209 amino acids) with multiple functions, including RNA silencing suppression (RSS). Confocal laser-scanning microscopy of green fluorescent protein (GFP)-p23 agroexpressed in Nicotiana benthamiana revealed its accumulation in the nucleolus, Cajal bodies, and plasmodesmata. To dissect the nucleolar localization signal (NoLS) typically associated with basic motifs, seven truncated and 10 point-mutated versions of p23 were assayed. Deletion mutants showed that regions 50 to 86 and 100 to 157 (excluding fragment 106 to 114), both with basic motifs and the first with a zinc-finger, contain the (bipartite) NoLS. Alanine substitutions delimited this signal to three cysteines of the Zn-finger and some basic amino acids. RSS activity of p23 in N. benthamiana was abolished by essentially all mutants, indicating that it involves most p23 regions. The necrotic-inducing ability of p23 when launched in N. benthamiana from Potato virus X was only retained by deletion mutant 158-209 and one substitution mutant, showing that the Zn-finger and flanking basic motifs form part of the pathogenic determinant. Ectopic expression of p23 and some deletion mutants in transgenic Mexican lime demarcated a similar determinant, suggesting that p23 affects related pathways in citrus and N. benthamiana. Both RSS activity and pathogenicity of p23 appear related to its nucleolar localization. PMID:23387469
Citrus variegated chlorosis (CVC), causal agent Xylella fastidiosa (Xf), huanglongbing (HLB) causal agent Liberobacter spp, and exotic stem pitting isolates of citrus tristezavirus (CTV) are not present in the continental U.S. CVC is transmitted by sharpshooter leafhoppers (Homoptera: Cicadellidae...
Citrus tristezavirus (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\\u000a of ten candidate Ctv resistance genes. The ten candidate genes were individually cloned in Agrobacterium based binary
In an attempt to utilize post-transcriptional gene silencing (PTGS) as a means to impart resistance against Citrus tristezavirus (CTV) into citrus plants, the p23 + 3?UTR sequence (p23U) of the VT strain of CTV was engineered to fold into a double-stranded\\u000a (ds) RNA structure. The resulting construct (p23UI) was introduced into Nicotiana benthamiana and Alemow (Citrus macrophylla) plants by Agrobacterium-mediated transformation.
Freshwater blue-green algae of the genera Lyngbya, Plectonema, and Phormidium are susceptible to a virus recently isolated from a waste-stabilization pond. Electron micrographs of a partially purified preparation show that the viral particle has an icosahedral structure about 66 mmu in diameter.
Grapefruit (Citrus paradisi Macf. cv Duncan) plants were transformed with several sequences from citrus tristeza closterovirus (CTV) that varied in terms of position in the CTV genome and virus strain origin in an attempt to obtain resistant plants. The sequences included the capsid protein gene from three different strains, a nontranslatable version of the capsid protein gene, the replicase (RdRp),
V. J. Febres; C. L. Niblett; R. F. Lee; G. A. Moore
Isolates of rinderpest virus (RPV) recovered from outbreaks of the disease in Kenya and Southern Sudan between 1986 and 1993 were compared to each other and to earlier isolates from East and West Africa. The recent isolates were mildly pathogenic for susceptible cattle and thus resembled other mild strains of RPV recovered from cattle and wildlife in East Africa more
Summary Cross hemagglutination inhibition (HI) and neuraminidase inhibition (NI) tests demonstrated strong antigenic relationship between A\\/Swine\\/Wisconsin\\/1\\/73 (SW\\/73) and A\\/Swine\\/Shope\\/15\\/31 (SW\\/31) influenza viruses. An eightyone fold purification of virus was achieved by adsorption and elution followed by differential ultracentrifugation and sedimentation through linear sucrose gradient. Radio-immunoassay using purified125I labeled viral antigens revealed antigenic variation between the two virusisolates. Neuraminidase of
Immunohistochemistry and virusisolation were performed on 1,057 birds. Immunohistochemistry, virusisolation, or both found 325 birds to be West Nile virus positive. Of these, 271 were positive by both methods. These results indicate that virusisolation and immunohistochemistry are approximately equal in their ability to detect West Nile virus.
Ellis, Angela E.; Mead, Daniel G.; Allison, Andrew B.; Gibbs, Samantha E. J.; Gottdenker, Nicole L.; Stallknecht, David E.; Howerth, Elizabeth W.
Expression of the RNA-dependent RNA polymerase (RdRp) of Citrus tristezavirus (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...
Between October 1993 and March 1994, outbreaks of pox-like exanthemas were observed in several camel raising farms in Dubai. Scabs from twenty camels with either local or generalized lesions were examined, seven of them had previously been vaccinated with a modified live camelpox virus vaccine. Inspection of scabs by electron microscopy confirmed an infection with orthopox viruses (OPV) in 10 animals and with parapox virus in one camel. Investigation of the scabs by polymerase chain reaction and dot blot assay revealed the presence of OPV in 15 or 13 samples, respectively. OPV could be isolated in cell culture in 14 cases. Restriction enzyme profiles characterized all isolates as camelpox virus. Their DNA patterns were virtually identical displaying only slight variations in the terminal fragments. In contrast, the vaccine strain showed a distinct restriction enzyme profile, indicating that it was not involved in the infections. PMID:8861650
Usutu virus (USUV) is a mosquito-borne flavivirus that emerged 2001 in Austria and caused deaths in wild birds. In Germany, 70,378 female mosquitoes were captured in 2009 and 2010 and assayed for USUV. Virus was isolated in cell culture from one pool of Culex pipiens pipiens mosquitoes trapped exclusively in August 2010 in Weinheim, Germany. Subsequent phylogenetic analysis demonstrated a close relationship between the isolated USUV strain from Germany and a USUV strain from Austria, which was detected in a dead blackbird in 2004.
Jost, Hanna; Bialonski, Alexandra; Maus, Deborah; Sambri, Vittorio; Eiden, Martin; Groschup, Martin H.; Gunther, Stephan; Becker, Norbert; Schmidt-Chanasit, Jonas
Usutu virus (USUV) is a mosquito-borne flavivirus that emerged 2001 in Austria and caused deaths in wild birds. In Germany, 70,378 female mosquitoes were captured in 2009 and 2010 and assayed for USUV. Virus was isolated in cell culture from one pool of Culex pipiens pipiens mosquitoes trapped exclusively in August 2010 in Weinheim, Germany. Subsequent phylogenetic analysis demonstrated a close relationship between the isolated USUV strain from Germany and a USUV strain from Austria, which was detected in a dead blackbird in 2004. PMID:21896821
Jöst, Hanna; Bialonski, Alexandra; Maus, Deborah; Sambri, Vittorio; Eiden, Martin; Groschup, Martin H; Günther, Stephan; Becker, Norbert; Schmidt-Chanasit, Jonas
A virus, which is morphologically identical and antigenically related to two previously known isolates of infectious bursal disease (IBD) virus, was isolated from pooled faeces of 6?week?old turkeys with diarrhoea. It is concluded that this virus, designated TY89, is an isolate of IBD virus. The isolation of TY89 was heavily dependent upon the use of electron microscopy and the immunofluorescence
Nucleotide sequence analysis was performed on 14 dengue virusisolates (13 dengue-2 viruses and 1 dengue-3 virus) recovered from febrile soldiers in Somalia in 1993. The dengue-2 viruses were most closely related to dengue-2 virus recovered in Somalia in 1984. However, differences in nucleotide sequence (0.35% to 1.35%) were evident among the 1993 isolates. These differences were closely associated with
Niranjan Kanesa-thasan; Gwong-Jen J. Chang; Bonnie L. Smoak; Alan Magill; M. Jeanne Burrous; Charles H. Hoke
African swine fever (ASF) is widespread in Africa but is rarely introduced to other continents. In June 2007, ASF was confirmed in the Caucasus region of Georgia, and it has since spread to neighboring countries. DNA fragments amplified from the genome of the isolates from domestic pigs in Georgia in 2007 were sequenced and compared with other ASF virus (ASFV) isolates to establish the genotype of the virus. Sequences were obtained from 4 genome regions, including part of the gene B646L that encodes the p72 capsid protein, the complete E183L and CP204L genes, which encode the p54 and p30 proteins and the variable region of the B602L gene. Analysis of these sequences indicated that the Georgia 2007 isolate is closely related to isolates belonging to genotype II, which is circulating in Mozambique, Madagascar, and Zambia. One possibility for the spread of disease to Georgia is that pigs were fed ASFV-contaminated pork brought in on ships and, subsequently, the disease was disseminated throughout the region.
Aquatic birds are the natural hosts for influenza virus. It is established that avian influenza viruses provide the gene pool for the generation of new strains of human influenza virus, which can cause pandemic infections. The recent outbreak of an avian influenza virus (H5N1) in Hong Kong not only produced high mortality in chickens, but also resulted in six human fatalities. This outbreak indicates that avian influenza virus can be pathogenic for humans. We surveyed local waterfowl habitats by taking water and fecal samples for virusisolation and identification. We isolated avian influenza viruses from ponds and small lakes in Bartlesville, Lawton, Stillwater, and Tulsa. The density of birds in these sites is small. However, our virusisolation rate is comparable to that found in higher density habitats. The risk of human infection remains to be determined. We encourage primary care physicians to submit samples for virus surveillance. PMID:10616258
Three strains of nephropathia epidemica (NE) virus were isolated from lung tissues of bank voles (Clethrionomys glareolus) and a grey-sided vole (C. rufocanus) trapped in Västerbotten county, Sweden. Two of these isolates were serially passaged in seronegative laboratory-bred bank voles. Experimentally infected animals developed a subclinical infection characterized by virus persistence, particularly in lung tissue. Attempts to infect other species of colonized rodents with NE virus and to isolate NE virus from acute phase patient blood were unsuccessful. The serial propagation of NE virus in colonized bank voles provides opportunities to study experimental infection in its reservoir rodent host. PMID:6208601
Yanagihara, R; Svedmyr, A; Amyx, H L; Lee, P; Goldgaber, D; Gajdusek, D C; Gibbs, C J; Nyström, K
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...
Outbreaks of an epidermic disease suggesting parapox virus infections have been observed in all major herds of sheep and goats from different geographical areas of Brazil. Clinical samples (dried scabs) were collected and orf virus was isolated and characterized by electron microscopy in previous work. In order to characterize these viruses at the molecular level, a modified methodology for genomic
C Mazur; I. I Ferreira; F. B Rangel Filho; R Galler
Five cases of orf virus infection in Korean black goats were diagnosed in our laboratory between 2010 and 2011. One orf virus (ORF/2011) was isolated from an ovine testis cell line (OA3.Ts) for use as a vaccine candidate. Sequences of the major envelope protein and orf virus interferon resistance genes were determined and compared with published reference sequences. Phylogenetic analyses revealed that orf viruses from Korean black goats were most closely related to an isolate (ORF/09/Korea) from dairy goats in Korea. This result indicates that the orf viruses might have been introduced from dairy goats into the Korean black goat population.
Oem, Jae-Ku; Chung, Joon-Yee; Kim, Yong-Joo; Lee, Kyoung-Ki; Kim, Seong-Hee; Jung, Byeong-Yeal
We have completely sequenced the genomes of two Nipah virus (NiV) isolates, one from the throat secretion and the other from the cerebrospinal fluid (CSF) of the sole surviving encephalitic patient with positive CSF virusisolation in Malaysia. The two genomes have 18246 nucleotides each and differ by only 4 nucleotides. The NiV genome is 12 nucleotides longer than the
Y. P. Chan; K. B. Chua; C. L. Koh; M. E. Lim; S. K. Lam
Tobacco streak virus (TSV) is an ilarvirus with a worldwide distribution. This virus infects many plants and causes significant yield losses. In this study, 300 samples of lettuce were collected from lettuce fields in Tehran Province. Infected plants show symptoms such as: mosaic, vein clearing, vein necrosis, yellowing and leaf distortion. DAS-ELISA (Double Antibody Sandwich-ELISA) was used with a polyclonal antiserum against TSV. Five isolates (T1, T2, T3, T4 and T5), which are collected, respectively from Mohammad Abad (Karaj), Malek Abad (Karaj), Hashtgerd (Karaj), Tarand Balla (Varamin) and Deh mah sin (Pishva) were inoculated on 29 species of Cucurbitaceae, Amaranthaceae, Solanacea, Compositae, Leguminosae and Chenopodiacea. Chenopodium quinoa 6 days after inoculation showed necrotic local lesions. Gomphrena globosa 10 days after inoculation developed chlorotic local lesions. Systemic symptoms were produced in Datura stramonium. Phaseolus vulgaris cv. Red Kidney 5 days after inoculation developed necrotic local lesions. Nicotiana tabacum 7 days after inoculation showed necrotic and chlorotic local lesions. Nicotiana clevelandii 15 days after inoculation developed leaf distortion and vein necrosis. Lactuca sativa 10-15 days after inoculation developed leaf istortion and mosaic. Reverse Transcription Polymerase Chain Reaction (RT-PCR) was performed using one primer pairs designed by DSMZ. An approximately 710 bp fragment was amplified with a specific primer. PMID:19634475
Summary. ?Isolates of infectious bursal disease virus (IBDV) were obtained from domestic poultry in New Zealand in 1997 and 1998. An\\u000a in-vivo pathogenicity study carried out in specific pathogen free (SPF) chickens demonstrated the low virulence of one of\\u000a the virusisolates. The nucleotide sequences of the hypervariable region of the VP2 gene of two isolates were determined and\\u000a compared with
Y. F. Chai; N. H. Christensen; C. R. Wilks; J. Meers
Banana bunchy top virus (BBTV) DNA component 1 from isolates from 10 different countries was cloned and sequenced and the sequences were aligned and com- pared. This analysis indicated two groups: the South Pacific group (isolates from Australia, Burundi, Egypt, Fiji, India, Tonga and Western Samoa) and the Asian group (isolates from the Philippines, Taiwan and Vietnam). The mean sequence
The comparative characteristics of biological, physico-chemical and antigenic properties of bean yellow mosaic virusisolated from soybeans are shown. The obtained results indicate the existence of individual properties of the isolate which help to distinguish it from the typical strains and isolates studied previously in Ukraine by other authors. PMID:23866589
Outbreaks of an epidermic disease suggesting parapox virus infections have been observed in all major herds of sheep and goats from different geographical areas of Brazil. Clinical samples (dried scabs) were collected and orf virus was isolated and characterized by electron microscopy in previous work. In order to characterize these viruses at the molecular level, a modified methodology for genomic DNA extraction directly from scabs was used and such DNA was used to derive the restriction enzyme digestion patterns for clinical samples from three distinct geographic origins. Pulsed field gel electrophoresis was used to separate restriction enzyme DNA fragments and heterogeneity among isolates from different geographic areas could be observed on stained gels. The HindIII-G DNA fragment from orf-A virus genome was cloned and hybridized to DNA of other orf virusisolates. Further heterogeneity was confirmed by these hybridizations. PMID:10781724
Mazur, C; Ferreira, I I; Rangel Filho, F B; Galler, R
In the course of studies on the ecological structure of acute febrile diseases in the season of activity of blood-sucking arthropods strains of a virus antigenically related to Sikhote-Alyń virus were isolated from the blood of a patient and from Ixodid ticks. This paper presents the results of the study on the causative agent and the clinical picture of the disease caused by this virus. The virus was found to be a new one for science; its appurtenance to the family Picornaviridae, genus Cardiovirus, the antigenic group of encephalomyocarditis has been determined. The virus has been designated "Syr-Darya Valley fever virus" by the area of its isolation. PMID:6097042
L'vov, D K; Karimov, S K; Kiriushchenko, T V; Chun-Siun, F; Skvortsova, T M
Momordica charantia L. plants systemically infected with Cucumber mosaic virus (CMV) were found in Oita Prefecture. The virusisolated from the host plant was characterized by biological, serological,\\u000a and molecular biological methods. The purified virus was used to mechanically inoculate the host and produced green mottle,\\u000a green mosaic, and\\/or chlorotic spots in the noninoculated upper leaves of the host. The
Record is made of the initial isolation and first 16 passages of the avian reticuloendotheliosis virus (strain T). The original infective liver tissue was taken from a moribund adult turkey having gross lesions similar to those of lymphoid leuosis. During...
A virusisolated in cell culture from the spleen of a cat with feline infectious peritonitis was identified by physicochemical, morphological and antigenic criteria as a coronavirus. The feline infectious peritonitis virus was compared in vitro with canine coronavirus, a reported enteric pathogen of dogs. The feline isolate was characterized, by chloroform sensitivity and resistance to 5-iododeoxyuridine, respectively, as containing essential lipid and an RNA genome. Other traits of the isolate included resistance to acidic conditions, heat lability, and resistance to trypsin. Electron microscopy showed viral particles with a structure consistent with that of the prototype of the coronavirus group, infectious bronchitis virus. Indirect immunofluorescence with canine coronavirus monospecific antiserum showed the viral isolate to be antigenically related to canine coronavirus. Specific-pathogen-free cats inoculated by various routes with cell-culture-propagated virus had both clinical symptoms and lesions consistent with feline infectious peritonitis. PMID:7467085
Evermann, J F; Baumgartener, L; Ott, R L; Davis, E V; McKeirnan, A J
A swine influenza H1N1 virus was isolated from a pig during a severe outbreak of respiratory disease in Korea. All genes of the H1N1 isolate, including hemagglutinin (HA), neuraminidase (NA), matrix (M), nucleoprotein (NP), non-structural (NS), PA, PB1 and PB2, were of swine origin. Also, all these genes showed a close phylogenic relationship with those of H1N1 viruses previously isolated
D. S. Song; C. S. Lee; K. Jung; B. K. Kang; J. S. Oh; Y. D. Yoon; J. H. Lee; B. K. Park
Background & objectives: Chittoor virus (CHITV) belongs to genus Orthobunyavirus, family Bunyaviridae. It has been isolated from various species of mosquitoes and pig from different parts of India. Five isolates of CHITV were characterized at the molecular level and compared with other Batai viruses (BATV) to find out any kind of reassortment in their genome. Methods: Complete nucelocapsid (S), glycoprotein (M) and partial RNA polymerase (L) segments of CHITV were amplified and sequenced. These sequences were compared with those of Batai viruses, isolated from different geographical locations in Asia, Africa and Europe. Results: Phylogenetic analysis revealed CHITV as a variant of BATV. High level of conservation was seen among the CHITV isolates studied. The CHITV sequences showed clustering in one lineage with the sequences from Japan and Malaysia, however, BATV sequences from Europe and Africa formed a separate phylogenetic lineage. Interpretation & conclusions: The study indicates the presence of a single genotype of CHITV circulating in India, despite the involvement of different hosts in the natural cycle by this virus. Analysis of the sequences of the S, M and L segments of genome indicated that the virus has not undergone any reassortment. This virus has not caused any epidemic involving humans, however, replication of the virus in different mosquito and vertebrate hosts species suggests that it is a cause of concern.
We describe herein a field case of border disease (BD) in twin lambs. Both lambs were unthrifty, stunted, and one exhibited nervous signs characteristic of BD, with tremors of the head, neck, hind legs, and pelvis. Hairiness of the coat and excessive pigmentation, often seen in lambs with BD, were not observed. A noncytopathic virus, which showed cross-reactivity with bovine viral diarrhea (BVD) virus antiserum and BVD virus monoclonal antibodies, was isolated repeatedly from leukocytes from one lamb and from tissues of the other. Although the source of the virus is unknown, our results suggest that the dam of the affected twins had been infected during pregnancy. We used the BD virusisolated to inoculate pregnant ewes and experimentally reproduce the disease in a newborn lamb. Our findings indicate that leukocytes, rather than serum, should be utilized for BD virusisolation. Further, it is recommended that BD virus, rather than BVD virus, be used in serum neutralization tests when screening sheep for antibody titers.
Lees, V. Wayne; Loewen, Ken G.; Deregt, Dirk; Knudsen, Robin
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.
Two rod-shaped viruses of similar length occurring in hops were isolated from a mixed infection. One (hop mosaic virus) infected Nicotiana clevelandii, produced mosaic symptoms in mosaic-sensitive hop cultivars, and did not infect Chenopodium murale. The other (hop latent virus) infected C. murale but not N. clevelandii and did not produce mosaic symptoms in hops. Neither N. clevelandii nor C. murale developed distinct symptoms after infection with the respective virus. Both viruses produced a systemic chlorotic flecking in Cluster hop seedling clones. PMID:986870
A multiplex polymerase chain reaction (mPCR) assay was developed to detect six RNA and one DNA citrus virus: Citrus leaf rugose virus (CLRV), Citrus psorosis virus (CPsV), Citrus tatter leaf virus (CTLV), Citrus tristezavirus (CTV), Citrus variegation virus (CVV), Citrus yellow mosaic virus (CYMV), and Indian citrus ringspot virus (ICRSV) from citrus plants. These seven viruses are classified in
Summary. A virusisolated from sorghum in Nigeria has been partially characterized. It was tested by enzyme-linked immunosorbent assay (ELISA) using antisera to Maize dwarf mosaic virus, Johnsongrass mosaic virus (JGMV), Sugarcane mosaic virus strain-MDB, Sorghum mosaic virus, and Zea mosaic virus. A partial host range, symptom phenotypes for selected sorghum lines, and the mass of the coat protein (CP)
D. L. Seifers; S. Haber; W. Ens; Y.-M. She; K. G. Standing; R. Salomon
Summary Transformed cells have been isolated from the bone marrow of reticuloendotheliosis virus (REV)-infected, moribund chicks. These cells have been maintained in vitro through more than 30 serial passages. The cells induce solid tumors when inoculated into the wing web of day-old chickens, and cell-free filtrates induce reticuloendotheliosis. Virus particles recovered after cocultivation of bone marrow cells and chicken embryo
Ray B. Franklin; Reynaldo L. Maldonado; Henry R. Bose
Multiple isolates of H5N1 were isolated from northern Vietnam in December of 2005. All the viruses characterized were clade 2 viruses, but phylogenetically they formed two separate sub-lineages. The isolation of clade 2 viruses was unexpected, since previous isolations from both northern and south...
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
An isolate of wisteria vein mosaic virus (WVMV) obtained fromWisteria sinensis in Prague resembled in its properties WVMV isolates described in Italy and Holland.Nicotiana megalosiphon is reported as a new host of WVMV. Other known host plants showed reactions similar to those described formerly. The incubation\\u000a period extended in some hosts up to two or four weeks. Pea plants showed
Genetic typing of 29 Bovine Viral Diarrhea Virus (BVDV) isolates from Argentina was carried out by sequencing 245 nucleotides of the RT-PCR products of the 5?-UTR region. Sequence analysis shows that these Argentinean BVDV include types 1 and 2. The majority (26\\/29) of the isolates are type 1, which comprises subtypes 1a and 1b, together with an additional subgroup within
Summary. ?Two new classical swine fever virus (CSFV) isolates obtained from naturally infected swine were found to exhibit a cytopathogenic\\u000a (cp) phenotype. According to their reactivity with monoclonal antibodies (mabs) the isolates cpBW1 and cpMVP1 were classified\\u000a as antigenic types “Lothringen’92” and “Flandern’90”, respectively. In Northern blot analyses and PCR assays CSFV RNA of subgenomic\\u000a length was detected in infected cells
IN February 1976, during an epidemic of influenza among military recruits at Fort Dix, New Jersey, five virusisolates were obtained which contained haemagglutinin and neuraminidase antigens indistinguishable from those of isolates of swine influenza virus1. Subsequent evidence of elevated serum antibody titre against `swine' influenza virus in hundreds of individuals at Fort Dix2 prompted the US government to launch
Wild aquatic birds are the primary reservoir of influenza A viruses, but little is known about the viruses' gene pool in wild birds. Therefore, we investigated the ecology and emergence of influenza viruses by conducting phylogenetic analysis of 70 matrix (M) genes of influenza virusesisolated from shorebirds and gulls in the Delaware Bay region and from ducks in Alberta,
Linda Widjaja; Scott L. Krauss; Richard J. Webby; Tao Xie; Robert G. Webster
An influenza A virus of the H7 N1 subtype was isolated from young ostriches which died after developing a syndrome characterized by a green discolouration of the urine, weakness and signs of respiratory distress. Mortality varied, depending on the age of the ostriches, the presence of other infectious agents and the amount of stress to which they were exposed. Using
D. M. Allwright; W. P. Burger; Adelaide Geyer; A. W. Terblanche
In India, rabies is enzootic and is a serious public health and economic problem. India has a large population of stray dogs which, together with a lack of effective control strategies, might have led to the persistence of rabies virus (RV) in the canine population. Our objective was to study the molecular epidemi- ology of RV isolates in India based
T. Nagarajan; B. Mohanasubramanian; E. V. Seshagiri; S. B. Nagendrakumar; M. R. Saseendranath; M. L. Satyanarayana; D. Thiagarajan; P. N. Rangarajan; V. A. Srinivasan
To compare the sensitivity and specificity of RT-PCR with that of virusisolation in the detection of human rhinoviruses, we tested nasopharyngeal aspirates from 200 patients on the 1st and 7th days after the onset of the common cold. An assay utilizing a short amplicon in the conserved 5* noncoding region was found highly sensitive. Of 192 positive samples altogether,
TIMO HYYPIA; TUOMO PUHAKKA; OLLI RUUSKANEN; MIKA MAKELA; ANITA AROLA; PERTTI ARSTILA
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...
Sequence analysis of part of the fusion protein gene from recent isolates of rinderpest virus revealed that distinct lineages of the virus exist which reflect the geographical location of their isolation in Africa and Asia. Current strains circulating in Kenya and Sudan were most similar, both in terms of nucleotide sequence and pathogenic nature, to virusesisolated in Egypt and
R. W. Chamberlain; H. M. Wamwayi; E. Hockley; M. S. Shaila; L. Goatley; N. J. Knowles; T. Barrett
When isolating dengue virus (DEN) from mosquitoes collected in endemic areas, pools may contain both anti-dengue antibodies from freshly engorged females and virus from DEN infected females. To determine if these antibodies may interfere with virusisolation, we simulated the isolation procedure using Aedes aegypti (L.) that we infected with the 16,681 strain of dengue type 2 virus by intrathoracic inoculation. At 7 d postinfection, we allowed females to engorge on immunized or normal mouse blood. Virus in a mixture of anti-dengue-2 antibodies and dengue-2 virus became inactive after incubation at 37 degrees C for 1 h, but remained infective without incubation. Therefore, at ambient conditions antibodies would not interfere with virusisolation from field-collected Ae. aegypti from endemic areas. In addition, DEN antibodies enhanced virus replication when inoculated into Ae. aegypti, but not C6/36 cells. The mechanism for this in vitro antibody enhancement of infection remains unclear. PMID:10916310
Porcine epidemic diarrhea virus (PEDV) LJB/03 strain was isolated from the feces of piglets suspected to be suffering from a severe diarrhea in Heilongjiang Province, and was identified by immunofluorescence test, immunelectronmicroscopy, RT-PCR and indirect ELISA assay. Characteristics of the virus culture and the methods of improvement of virus titer were explored. The results showed that the virus had the typical appearance of the coronavirus. Analysis of the nucleotide sequences of RT-PCR products revealed 98% homology with the reference strains. Indirect immunofluorescence assay showed a significant presence of green fluorescence, and an average P/N ratio of 7.6 by indirect ELISA assay. Taken together, these tests showed positive isolation of PEDV. Using the virus plaque purification cloning methods established in the test, the purified PEDV large plaque and small plaque were obtained, and the large plaque and small plaque titers were measured with significant difference. These results provide potential for the application of PEDV on the basis of the biological features of isolatedvirus. PMID:21344754
A virusisolated from dead Chaerephon plicata bats collected near Kampot, Cambodia, was identified as a member of the family Bunyaviridae by electron microscopy. The only bunyavirus previously isolated from Chaerephon species bats in South-East Asia is Kaeng Khoi (KK) virus (genus Orthobunyavirus), detected in Thailand over 30 years earlier and implicated as a public health problem. Using RT-PCR, nucleotide sequences from the M RNA segment of several virusisolates from the Cambodian C. plicata bats were found to be almost identical and to differ from those of the prototype KK virus by only 2.6-3.2 %, despite the temporal and geographic separation of the viruses. These results identify the Cambodian bat viruses as KK virus, extend the known virus geographic range and document the first KK virusisolation in 30 years. These genetic data, together with earlier serologic data, show that KK viruses represent a distinct group within the genus Orthobunyavirus. PMID:13679602
Osborne, J C; Rupprecht, C E; Olson, J G; Ksiazek, T G; Rollin, P E; Niezgoda, M; Goldsmith, C S; An, U S; Nichol, S T
Summary Outbreaks of swine influenza were first observed in Japan in 1978. A number of influenza viruses were isolated from diseased swine. Almost all virusesisolated were swine influenza virus (Hsw1N1) but two virusesisolated from the nasal swabs of swine showing clinical signs of influenza in the Kanagawa prefecture were characterized antigenically as Hsw1N2. Analysis of swine sera showed
T. Sugimura; H. Yonemochi; T. Ogawa; Y. Tanaka; T. Kumagai
H3N3 and H1N1 influenza A viruses were isolated from Canadian pigs in 2001 and 2002. These viruses are phylogenetically related to waterfowl viruses and antigenically distinct from reference swine influenza viruses. The isolation of these viruses reemphasizes the potential for interspecies transmission of influenza viruses from waterfowl to pigs in North America.
Karasin, Alexander I.; West, Keith; Carman, Suzanne; Olsen, Christopher W.
Bluetongue (BT) is an economically important endemic disease of livestock in tropics and subtropics. In addition, its recent spread to temperate regions like North America and Northern Europe is of serious concern. Rapid serotyping and characterization of BT virus (BTV) is an essential step in the identification of origin of the virus and for controlling the disease. Serotyping of BTV is typically performed by serum neutralization, and of late by nucleotide sequencing. This report describes the near complete genome sequencing and typing of two isolates of BTV using Illumina next generation sequencing platform. Two of the BTV RNAs were multiplexed with ten other unknown samples. Viral RNA was isolated and fragmented, reverse transcribed, the cDNA ends were repaired and ligated with a multiplex oligo. The genome library was amplified using primers complementary to the ligated oligo and subjected to single and paired end sequencing. The raw reads were assembled using a de novo method and reference-based assembly was performed based on the contig data. Near complete sequences of all segments of BTV were obtained with more than 20× coverage, and single read sequencing method was sufficient to identify the genotype and serotype of the virus. The two viruses used in this study were typed as BTV-1 and BTV-9E. PMID:23831448
Banana bunchy top virus (BBTV) DNA component 1 from isolates from 10 different countries was cloned and sequenced and the sequences were aligned and compared. This analysis indicated two groups: the South Pacific group (isolates from Australia, Burundi, Egypt, Fiji, India, Tonga and Western Samoa) and the Asian group (isolates from the Philippines, Taiwan and Vietnam). The mean sequence difference within each group was 1.9 to 3.0% and between isolates from the two groups was approximately 10%, but some parts of the sequences differed more than others. However, the protein encoded by the major open reading frame, which is probably a replicase, differed by approximately 5%. The region from the beginning of the stem-loop sequence to the potential TATA box was identical in all isolates except for a two nucleotide change in the Western Samoan isolate and a single change in that of the NSW isolate. These results, together with other evidence, suggest that BBTV has spread to bananas after the initial movement of bananas from the Asian Pacific regions to Africa and the Americas. PMID:7996145
Summary A virus with rhabdovirus morphology which proved to be antigenically distinct from rabies virus and vesicular stomatitis virus was isolated from a dolphin that had beached on the Dutch coast. Neutralizing antibodies to this virus were found in several European marine mammal species.
A. D. M. E. Osterhaus; H. W. J. Broeders; J. S. Teppema; T. Kuiken; J. A. House; H. W. Vos; I. K. G. Visser
The genus Henipavirus in the family Paramyxoviridae contains two viruses, Hendra virus (HeV) and Nipah virus (NiV) for which pteropid bats act as the main natural reservoir. Each virus also causes serious and commonly lethal infection of people as well as various species of domestic animals, however little is known about the associated mechanisms of pathogenesis. Here, we report the isolation and characterization of a new paramyxovirus from pteropid bats, Cedar virus (CedPV), which shares significant features with the known henipaviruses. The genome size (18,162 nt) and organization of CedPV is very similar to that of HeV and NiV; its nucleocapsid protein displays antigenic cross-reactivity with henipaviruses; and it uses the same receptor molecule (ephrin- B2) for entry during infection. Preliminary challenge studies with CedPV in ferrets and guinea pigs, both susceptible to infection and disease with known henipaviruses, confirmed virus replication and production of neutralizing antibodies although clinical disease was not observed. In this context, it is interesting to note that the major genetic difference between CedPV and HeV or NiV lies within the coding strategy of the P gene, which is known to play an important role in evading the host innate immune system. Unlike HeV, NiV, and almost all known paramyxoviruses, the CedPV P gene lacks both RNA editing and also the coding capacity for the highly conserved V protein. Preliminary study indicated that CedPV infection of human cells induces a more robust IFN-? response than HeV.
Treatment of a cell line derived from the Asian feral mouse Mus caroli with 5-bromodeoxyuridine induces an infectious, xenotropic type C virus. This virus shares strongly cross-reactive reverse transcriptase (RNA-dependent DNA polymerase) and p30 antigens and cross-interferes with type C virusesisolated from a woolly monkey (SSAV) and gibbon apes (GALV). By similar criteria, the caroli virus is much less
Michael M. Lieber; Charles J. Sherr; George J. Todaro; Raoul E. Benveniste; Robert Callahan; Hayden G. Coon
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
Eight Newcastle disease virusisolates from Pakistan were sequenced and characterized. A PCR matrix gene assay, designed to detect all avian paramyxovirus 1, did not detect four of the isolates. A new matrix gene test that detected all isolates was developed. Phylogenetic analysis and pathotyping confirmed that virulent viruses of different genotypes are circulating in Pakistan. PMID:20237105
Khan, Taseer Ahmed; Rue, Cary A; Rehmani, Shafqat Fatima; Ahmed, Ayaz; Wasilenko, Jamie L; Miller, Patti J; Afonso, Claudio L
In 2010, H14 influenza A viruses were recovered from clinically normal sea ducks in the United States. These are the first H14 isolates recovered in the Western Hemisphere and represent the only documented H14 influenza A virusesisolated since the original isolates were recovered from near the Caspian Sea during 1982. PMID:22307173
Nolting, Jacqueline; Fries, Anthony C; Slemons, Richard D; Courtney, Chad; Hines, Nichole; Pedersen, Janice
We report here the complete genome sequence of the porcine epidemic diarrhea virus strain CH/ZMDZY/11 isolated from central China. Our data, together with sequence data of porcine epidemic diarrhea virus (PEDV) isolates from other parts in China, will help to understand better the epidemiology and genetic diversity of PEDV field isolates in China.
We report here the complete genome sequence of the porcine epidemic diarrhea virus strain CH/ZMDZY/11 isolated from central China. Our data, together with sequence data of porcine epidemic diarrhea virus (PEDV) isolates from other parts in China, will help to understand better the epidemiology and genetic diversity of PEDV field isolates in China. PMID:23469356
During 1990, Dengue-2 (DEN-2) virus was isolated for the first time from mosquitoes (Aedes furcifer, six isolates; Ae. taylori, six isolates; Ae. luteocephalus, seven isolates) collected during an epidemic in which DEN-2 virus also was isolated from humans. Numerous isolations have been made previously from mosquitoes in the absence of human infection. In Senegal, DEN-2 virus appears to be maintained in an enzootic cycle and, therefore, plays an expanding role in human disease and increases the need for effective surveillance in mosquito populations. PMID:7932611
Traore-Lamizana, M; Zeller, H; Monlun, E; Mondo, M; Hervy, J P; Adam, F; Digoutte, J P
Summary One strain of Bhanja virus was isolated fromDermacentor marginatus male ticks collected from sheep in Slovak Karst, eastern Czechoslovakia (48°31'N, 20°28'E). In addition, three strains of Brezová virus (subtype of Tribec orbivirus,Reoviridae) were isolated fromIxodes ricinus ticks. The result represents the northernmost isolation of Bhanja virus and, moreover, its first recovery fromD. marginatus in Europe. Cross-neutralization plaque reduction test
Z. Hubfilek; T. Mittermayer; J. Halouzka; V. ?erný
In 2006, a swine influenza virus (SIV) isolate was isolated from 30 nasal swabs samples collected from pigs with clinical syndromes of swine influenza in a pig farm of Liaoning Province. The virusisolate was studied and identified by the growth in 9-11 days old chicken embryo, hemagglutination (HA) assay, hemagglutination inhibition (HI) assay, reverse transcription-polymerase chain reaction assays (RT-PCR) for its genetic subtype, whole gene sequence analysis and animal trial for its virulence. The virusisolate demonstrated the hemagglutination activity. Result of HI test against H1 subtype of SIV was positive, however, the results were negative when the HI studies were conducted using SIV H3 subtype virus and Newcastle Disease Virus (NDV). Eight gene segments of the virusisolate were amplified by RT-PCR. Phylogenetic analysis of the gene sequence of the virusisolate by using DNAstar software program revealed that the isolate have the H1 HA gene, by comparing to the sequences of H1-H16 in the GenBank. Furthermore, sequencing results also demonstrated that the virusisolate's NA gene belongs to N1 subtype. Therefore, the subtype of the SIV isolate is H1N1. The results of sequence analysis indicated that the genome of the SIV-H1N1 LN strain includes 8 fragments, among which only M protein gene is not swine originated. All other 7 fragments have close relationship with the domestic standard swine H1N1 strains. Results suggested that the SIV isolate LN strain might be created by a possible triple reassortants among the classic swine influenza virus, human influenza-like virus, and avian influenza-like virus. Piglets were inoculated with the SIV LN strain virus preparations and the virus caused the typical clinical symptoms of swine influenza in the inoculated piglets. This study, the isolation, identification and genetic analysis of the SIV LN strain provided useful information and basic data for the further investigation of epidemic principles and patterns of swine influenza virus in China. PMID:21043141
Since it was first described in Australia in 1994, Hendra virus (HeV) has caused two outbreaks of fatal disease in horses and humans, and an isolated fatal horse case. Our preliminary studies revealed a high prevalence of neutralizing antibodies to HeV in bats of the genus Pteropus, but it was unclear whether this was due to infection with HeV or
K. Halpin; P. L. Young; H. E. Field; J. S. Mackenzie
SUMMARY A small iridescent virus (type 29) has been isolated from the meal worm Tenebrio molitor. The virus is distinct from a number of previous isolates of small iridescent viruses (types 2, 6, 21, 22, 23 and 28) judged by polyacrylamide gel electro- phoresis of its structural polypeptides. Immunodiffusion and immunoprecipitation tests showed that iridescent virus type 29 is related
D. C. Kelly; M. D. Ayres; THELMA LESCOTT; J. S. Robertson; G. M. Happ
Nucleotide sequencing and phylogenetic analysis of Brazilian bovine viral diarrhea virus (BVDV) field isolates identified four viruses belonging to the genotype 2. Comparison of 5? UTR sequences from these isolates to those of North American BVDV type 2 revealed genomic variations that correlated with the geographic origins of the isolates. Two of the Brazilian type 2 viruses were isolated from
E. F Flores; L. H. G. V Gil; S. A Botton; R Weiblen; J. F Ridpath; L. C Kreutz; C Pilati; D Driemeyer; V Moojen; A. C Wendelstein
Between 1981 and 1983 10 isolations of infectious bronchitis (IB) virus were made from broiler chickens with respiratory infection and five from breeder chickens showing aberrant egg production. One isolate was serologically similar to an IB virusisolated in England in 1976; the remainder were related to three of the four serotypes of IB virus first isolated in The Netherlands
Infectious bronchitis (IB) is one of the important viral diseases of chickens, and in spite of regular vaccination, IB is a continuous problem in Canadian poultry operations. In an earlier study using sentinel chickens we determined the incidence of infectious bronchitis virus (IBV) in Ontario commercial layer flocks. The objective of this study was to determine the pathogenicity of 5 nonvaccine-related IBV isolates recovered from the sentinel birds. The clinical signs, gross, and histological lesions in specific pathogen-free chickens indicated that all 5 isolates caused mild lesions in the respiratory tract. An important finding of this study was the significantly lower average daily weight gain among virus-inoculated groups of chickens during the acute phase of infection. Based on sequences of part of the S1 gene IBV-ON2, IBV-ON3, and IBV-ON5 formed a cluster and they were closely related to strain CU-82792. IBV-ON4 had 98.7% identity with the strain PA/1220/9, a nephropathogenic variant.
Grgiae, Helena; Hunter, D. Bruce; Hunton, Peter; Nagy, Eva
SUMMARY Virusesisolated from the blood of two Korean haemorrhagic fever patients were propagated in cell culture and compared to prototype Hantaan virus which was isolated from Apodemus mice. The antigenic properties of the human isolates were found to be closely related to Hantaan virus by plaque reduction neutralization, haemagglutination inhibition and fluorescent antibody staining with both polyclonal and monoclonal
CONNIE S. SCHMALJOHN; JIRO ARIKAWA; SHERMAN E. HASTY; LYNN RASMUSSEN; HO WANG LEE; PYUNG WOO LEE; JOEL M. DALRYMPLE
Of the Orthomyxoviridae family of viruses, only influenza A viruses are thought to exist as multiple subtypes and has non-human maintenance hosts. In April 2011, nasal swabs were collected for virusisolation from pigs exhibiting influenza-like illness. Subsequent electron microscopic, biochemical, and genetic studies identified an orthomyxovirus with seven RNA segments exhibiting approximately 50% overall amino acid identity to human influenza C virus. Based on its genetic organizational similarities to influenza C viruses this virus has been provisionally designated C/Oklahoma/1334/2011 (C/OK). Phylogenetic analysis of the predicted viral proteins found that the divergence between C/OK and human influenza C viruses was similar to that observed between influenza A and B viruses. No cross reactivity was observed between C/OK and human influenza C viruses using hemagglutination inhibition (HI) assays. Additionally, screening of pig and human serum samples found that 9.5% and 1.3%, respectively, of individuals had measurable HI antibody titers to C/OK virus. C/OK virus was able to infect both ferrets and pigs and transmit to naive animals by direct contact. Cell culture studies showed that C/OK virus displayed a broader cellular tropism than a human influenza C virus. The observed difference in cellular tropism was further supported by structural analysis showing that hemagglutinin esterase (HE) proteins between two viruses have conserved enzymatic but divergent receptor-binding sites. These results suggest that C/OK virus represents a new subtype of influenza C viruses that currently circulates in pigs that has not been recognized previously. The presence of multiple subtypes of co-circulating influenza C viruses raises the possibility of reassortment and antigenic shift as mechanisms of influenza C virus evolution.
Hause, Ben M.; Ducatez, Mariette; Collin, Emily A.; Ran, Zhiguang; Liu, Runxia; Sheng, Zizhang; Armien, Anibal; Kaplan, Bryan; Chakravarty, Suvobrata; Hoppe, Adam D.; Webby, Richard J.; Simonson, Randy R.; Li, Feng
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
Zhou, Ming; Zhou, Zutao; Kia, Grace S N; Gnanadurai, Clement W; Leyson, Christina M; Umoh, Jarlath U; Kwaga, Jacob P; Kazeem, Haruna M; Fu, Zhen F
The 3 noncoding region (3 NCR) of flaviviruses contains secondary and tertiary structures essential for virus replication. Previous studies of yellow fever virus (YFV) and dengue virus have found that modifications to the 3 NCR are sometimes associated with attenuation in vertebrate and\\/or mosquito hosts. The 3 NCRs of 117 isolates of South American YFV have been examined, and major
Juliet E. Bryant; Pedro F. C. Vasconcelos; Rene C. A. Rijnbrand; J. P. Mutebi; Stephen Higgs; Alan D. T. Barrett
We analyzed the phylogenetic tree of densoviruses isolated from indigenous mosquitoes and mosquito cell lines. Our findings suggest two distinct clades of densovirus. The viruses in the first clade were isolated from an indigenous mosquito which had the Aedes aegypti densovirus (AaeDNV) as a representative virus. The other clade of viruses was isolated from mosquito indigenous cell line which had the Aedes albopictus densovirus (AalDNV) as the representative virus. The origin of the two clades of DNVs is unclear but the phylogenetic trees were significantly different from each other. The two major densoviruses, AaeDNV and AalDNV, that infect mosquitoes that are known to carry viruses responsible for dengue hemorrhagic fever and yellow fever. Understanding the evolution of these two clades of densoviruses is important for studying the distribution of these viruses in mosquito cell lines and the information gained may be applied to understanding other viruses in various mosquito cell lines. PMID:24050091
A noncytopathic type 1a bovine viral diarrhea virus (BVDV) was isolated from a free- ranging yearling female mule deer (Odocoileus hemionus) from northwestern Wyoming (USA). The mule deer was emaciated, weak, and salivating, and Arcanobacterium pyogenes was cultured from lung abscesses. Bovine viral diarrhea virus was isolated from lung, however, BVDV antigen was not detected by immunohistochemistry. The BVDV genotype
Hana Van Campen; Julia Ridpath; Elizabeth Williams; Jacqueline Cavender; Joan Edwards; Scott Smith; Hall Sawyer
The isolation of West Nile virus from the brain tissue of three children who died of encephalitis in Mysore and Kolar districts of Karnataka State, India, is reported. This is the first such report from India. The significance of these isolations with reference to the role of West Nile virus in encephalitis in children in southern India is discussed.
George, Samuel; Gourie-Devi, M.; Rao, J. A.; Prasad, S. R.; Pavri, K. M.
Technical Abstract A new virus named Nootka lupine vein-clearing virus (NLVCV) was isolated from native lupine plants confined to a relatively small area in the Talkeetna Mountains of south-central Alaska. Spherical particles about 30 nm in diameter were isolated from these leaves. Virions contained...
Commercial poultry farms in 14 regions of Russian Federation were monitored for avian leukosis virus (ALV) infection using virusisolation tests and serology. Results indicated the presence of two subgroups of ALV in farms located in 11 of 14 regions. Analysis of the genomes of 12 field isolates of...
Bovine viral diarrhea viruses (BVDV) were isolated from free-ranging bighorn sheep afflicted with pneumonia in late fall and winter of 2002. One of three bighorn ewes found dead was examined for gross lesions and samples were submitted for histopathology, aerobic bacterial culture, FA and virusisol...
Classical swine fever (CSF), a highly contagious viral disease of pigs, is endemic in India. As there is no information concerning the accurate genetic typing of classical swine fever virus (CSFV) isolates in India, 16 CSF virusesisolated during 2005–2007 from domestic pigs in different districts of Assam were typed in 5? UTR (150 nucleotides). To confirm the genetic typing
To determine the lineage of West Nile virus that caused outbreaks in Italy in 2008 and 2009, several West Nile virus strains were isolated from human specimens and sequenced. On the basis of phylogenetic analyses, the strains isolated constitute a distinct group within the western Mediterranean cluster.
Rossini, Giada; Carletti, Fabrizio; Bordi, Licia; Cavrini, Francesca; Gaibani, Paolo; Landini, Maria P.; Pierro, Anna; Capobianchi, Maria R.; Di Caro, Antonino
Summary An isolate of banana streak virus (BSV) that does not also occur as an integrant in the Musa balbisiana genome was sought in order to investigate the biological role of BSV in the evolution of either the Musa genome or of the virus itself. We isolated BSV virions from a Musa acuminata siamea accession from Vietnam and sequenced the entire
F. Lheureux; N. Laboureau; E. Muller; B. E. L. Lockhart; M.-L. Iskra-Caruana
Thirteen orf virusisolates obtained during the time period between 1995 and 2004 from crusted scab lesions of nine sheep and four goats from different geographical areas of Greece and Italy with suspected contagious ecthyma infection were analyzed. DNA of all isolates was successfully amplified by PCR with the primers 045F-045R and identified them as parapox virus. Partial DNA sequence
Christine Kottaridi; Kyriaki Nomikou; Liana Teodori; Giovanni Savini; Rossella Lelli; Panayotis Markoulatos; Olga Mangana
Strawberry latent ringspot virus (SLRV) was isolated from flowering cherry (Prunus serrulata sensu lato) trees in close proximity to each other in Auckland, New Zealand. The virus was not isolated from any of 390 flowering cherry\\u000a trees tested from four other regions in the North Island. The virus was identified by host range, particle morphology, RNA\\u000a and protein content and
1. A pneumotropic virus which forms elementary bodies has been isolated from apparently normal albino Swiss mice. 2. The antigenic relationship of this virus to those of meningopneumonitis, lymphogranuloma venereum, hamster pneumonia (7), and human pneumonitis (8) was established either by cross-immunity or complement fixation or both. 3. In spite of a relationship to other viruses, the virus could be differentiated from all the others studied by certain of its properties. PMID:19871384
Aleutian mink disease virus (AMDV) is a parvovirus that causes an immune complex mediated disease in minks. To understand the genetic characterization of AMDV in China, the genomic sequences of three isolates, ADV-LN1, ADV-LN2, and ADV-LN3, from different farms in the Northern China were analyzed. The results showed that the lengths of genomic sequences of three isolates were 4,543, 4,566, and 4,566 bp, respectively. They shared only 95.5-96.3 % nucleotide identity with each other. The nucleotide and amino acid homology of genome sequence between the Chinese isolates and European or American strains (ADV-G, ADV-Utah1, and ADV-SL3) were 92.4-95.0 % and 92.1-93.8 %, respectively. The amino acid substitutions randomly distributed in the genome, especially NS gene. ADV-LN1 strain had a 9-amino-acid deletion at amino acid positions 70 and 72-79 in the VP1 gene, comparing with ADV-G strain; ADV-LN2 and ADV-LN3 strains had 1-amino-acid deletion at amino acid positions 70 in the VP1. Some potential glycosylation site mutations in VP and NS genes were also observed. Phylogenetic analysis results showed that the three strains belonged to two different branches based on the complete coding sequence of VP2 gene. However, they all were in the same group together with the strains from United States based on the NS1 sequence. It indicated that Chinese AMDV isolates had genetic diversity. The origin of the ancestors of the Chinese AMDV strains might be associated with the American strains. PMID:22415541
Summary A study has been made of the use of chicken tracheal organ cultures for the isolation and assay of avian infectious bronchitis (AIB) virus from both naturally and experimentally infected chickens. Six strains of AIB virus were investigated, 3 of which had been isolated from natural outbreaks of disease. Two of the virusisolations from the outbreaks of AIB
A reverse transcription-PCR (RT-PCR) assay was developed and compared with enzyme immunoassay (EIA) and virusisolation for detecting LaCrosse virus (LAC) in mosquito pools. All three techniques were able to detect a single LAC-infected mosquito in a pool of 99 negative mosquitoes. Virusisolation was the most sensitive of the three techniques; it was possible to isolatevirus immediately following intrathoracic inoculation of mosquitoes. RT-PCR was second in sensitivity; LAC RNA was detected 1 day postinfection. EIA detected LAC antigen 2 days postinfection. Additionally, RT-PCR and EIA were able to detect LAC RNA and protein, respectively, from mosquito samples which were subjected to seven freeze-thaw cycles, and RT-PCR was able to detect LAC RNA from mosquito samples which remained at room temperature for up to 7 days. PMID:7814527
Wasieloski, L P; Rayms-Keller, A; Curtis, L A; Blair, C D; Beaty, B J
The complete 4463 nucleotide sequence for the medium segment viral RNA of Cache Valley virus has been cloned and sequenced in four isolates; in addition, the G1 glycoprotein extracellular coding domains are completed for nine additional isolates, including two subtypes, Ft. Sherman (86MSP18) and Tlacotalpan (61D240) viruses. The 13 represent isolations spanning over 45 years and a large geographic area,
Summary. 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
Summary Four isolates of infectious bursal disease virus (IBDV), isolated from chicken, duck, goose and sparrow in Jiangsu province\\u000a of China in 2002, were compared. The viruses were stable to the treatments of 60 °C for 1 h, pH 2.0 and lipid solvents. Their\\u000a antigenic relatedness values (R) were from 0.76 to 0.78. Chickens infected with the chicken isolate showed severe
Y. S. Wang; Z. C. Wang; Y. D. Tang; Z. L. Shi; K. W. He; Y. Li; J. B. Hou; H. C. Yao; H. J. Fan; C. P. Lu
During routine arbovirus surveillance from 2000 to 2004 in New York state (NYS), 14,788 mosquito pools making up 36 species and nine genera were inoculated onto Vero cell cultures to test for abroad spectrum of viruses. Forty-six percent of virusesisolated in cell culture from species, excluding Culex pipiens L. and Culex restuans Theobald, were identified as Bunyamwera serogroup viruses. Here, we report the distribution and level of Bunyamwera activity in NYS detected during this period. We developed specific primers for Cache Valley virus (family Bunyaviridae, genus Orthobunyavirus, CVV) and Potosi virus (family Bunyaviridae, genus Orthobunyavirus, POTV), to facilitate rapid molecular identification of these viruses. Viral RNA was detected in 12 mosquito species by reverse transcription-polymerase chain reaction, with the majority isolated from Aedes trivittatus (Coquillet). We report the first POTV isolation in NYS and describe the development of specific primers to identify both POTV and CVV. PMID:17017240
Ngo, Kiet A; Maffei, Joseph G; Dupuis, Alan P; Kauffman, Elizabeth B; Backenson, P Bryon; Kramer, Laura D
The genus Henipavirus in the family Paramyxoviridae contains two viruses, Hendra virus (HeV) and Nipah virus (NiV) for which pteropid bats act as the main natural reservoir. Each virus also causes serious and commonly lethal infection of people as well as various species of domestic animals, however little is known about the associated mechanisms of pathogenesis. Here, we report the
Glenn A. Marsh; Carol de Jong; Jennifer A. Barr; Mary Tachedjian; Craig Smith; Deborah Middleton; Meng Yu; Shawn Todd; Adam J. Foord; Volker Haring; Jean Payne; Rachel Robinson; Ivano Broz; Gary Crameri; Hume E. Field; Lin-Fa Wang
Since October 2001, the Adolfo Lutz Institute has been receiving vesicular fluids and scab specimens of patients from Paraíba Valley region in the Săo Paulo and Minas Gerais States and from Săo Patricio Valley, in the Goiás State. Epidemiological data suggested that the outbreaks were caused by Cowpox virus or Vaccinia virus. Most of the patients are dairy milkers that had vesiculo-pustular lesions on the hands, arms, forearms, and some of them, on the face. Virus particles with orthopoxvirus morphology were detected by direct electron microscopy (DEM) in samples of 49 (66.21%) patients of a total of 74 analyzed. Viruses were isolated in Vero cell culture and on chorioallantoic membrane (CAM) of embryonated chicken eggs. Among 21 samples submitted to PCR using primers for hemagglutinin (HA) gene, 19 were positive. Restriction digestion with TaqI resulted in four characteristic Vaccinia virus fragments. HA nucleotide sequences showed 99.9% similarity with Cantagalo virus, described as a strain of Vaccinia virus. The only difference observed was the substitution of one nucleotide in the position 616 leading to change in one amino acid of the protein in the position 206. The phylogenetic analysis showed that the isolates clustered together with Cantagalo virus, other Vaccinia strains and Rabbitpox virus. PMID:15654477
Nagasse-Sugahara, Teresa Keico; Kisielius, Jonas José; Ueda-Ito, Marli; Curti, Suely Pires; Figueiredo, Cristina Adelaide; Cruz, Aurea Silveira; Silva, Maysa Madalena J; Ramos, Carmen Helena; Silva, Maria Claudia C; Sakurai, Tiyo; Salles-Gomes, Luis Floręncio
An avian influenza virus (AIV), A/turkey/Israel/09 subtype H6N1, was isolated from turkey poults exhibiting typical pathology associated with AIV infection. The virus was characterized by RT-PCR using AIV subtype-specific primers and by the haemagglutination inhibition test using AIV subtype-specific antisera. The virus has an intravenous pathogenicity index of 0 and possessed a nucleotide sequence at the cleavage site of the hemagglutinin gene, PQIETR*GLF, associated with avian influenza viruses of low pathogenicity. Unlike the two previous H6N2 isolates originating from domestic ducks and mallard, the A/turkey/Israel/09 (H6N1) was isolated from turkeys. The gene sequences of the A/turkey/Israel/09 (H6N1) virus show divergence from the former Israeli H6 isolates. PMID:23074655
As part of a comprehensive study on the ecology of arthropod-borne viruses in the Amazon Basin region of Peru, we assayed 539,694 mosquitoes captured in Loreto Department, Peru, for arboviruses. Mosquitoes were captured either by dry ice-baited miniature light traps or with aspirators while mosquitoes were landing on human collectors, identified to species, and later tested on Vero cells for virus. In total, 164 virusisolations were made and included members of the Alphavirus (eastern equine encephalomyelitis, Trocara, Una, Venezuelan equine encephalomyelitis, and western equine encephalomyelitis viruses), Flavivirus (Ilheus and St. Louis encephalitis), and Orthobunyavirus (Caraparu, Itaqui, Mirim, Murutucu, and Wyeomyia viruses) genera. In addition, several viruses distinct from the above-mentioned genera were identified to the serogroup level. Eastern equine encephalomyelitis virus was associated primarily with Culex pedroi Sirivanakarn & Belkin, whereas Venezuelan equine encephalomyelitis virus was associated primarily with Culex gnomatos Sallum, Huchings & Ferreira. Most isolations of Ilheus virus were made from Psorophora ferox (Von Humboldt). Although species of the Culex subgenus Melanoconion accounted for only 45% of the mosquitoes collected, 85% of the virusisolations were made from this subgenus. Knowledge of the viruses that are being transmitted in the Amazon Basin region of Peru will enable the development of more effective diagnostic assays, more efficient and rapid diagnoses of clinical illnesses caused by these pathogens, risk analysis for military/civilian operations, and development of potential disease control measures. PMID:16366001
Turell, M J; O'Guinn, M L; Jones, J W; Sardelis, M R; Dohm, D J; Watts, D M; Fernandez, R; Travassos da Rosa, A; Guzman, H; Tesh, R; Rossi, C A; Ludwig, V; Mangiafico, J A; Kondig, J; Wasieloski, L P; Pecor, J; Zyzak, M; Schoeler, G; Mores, C N; Calampa, C; Lee, J S; Klein, T A
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 virusesisolated 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, virusisolated 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 virusesisolated from CNS in both macaques compared to SIVsmE543-3. Moreover, virusisolated from the brain in H631, had statistically significant loss of PNGS compared to virusisolated from CSF and plasma of the same animal. It is possible that the brain isolate may have adapted to decrease the number of PNGS given that humoral immune selection pressure is less likely to be encountered in the brain. These viruses provide a relevant model to study the adaptations required for SIV to induce encephalitis.
Foley, Brian T [Los Alamos National Laboratory; Korber, Bette T [Los Alamos National Laboratory
During the early spring of 1979 turkeys on at least twelve sites in England became infected with influenza A viruses. On five of these sites no virus was isolated but birds were shown to have antibodies to Havl (four sites) and Hav2 antigenic subtypes of influenza A viruses. The eight virusesisolated were typed:A\\/turkey\\/England\\/192–328\\/79 (Havl Nav2\\/3), A\\/turkey\\/England\\/192–329\\/79 (Hav1 N2), A\\/turkey\\/England\\/199\\/79
A feline panleukopenia virus (FPV) mutant, monkey/BJ-22/2008/CHN, was isolated from intestinal contents of a diarrheic monkey in Beijing, China. The virus was identified by morphology and physicochemical characteristics, and specific fragments were obtained by PCR using consensus primers of parvovirus and specific primers of FPV. Sequence of the full-length VP2 gene of the isolated FPV was determined and analyzed by comparison with reference FPV and canine parvovirus (CPV) isolates, showing high homology with FPV (98.75%) and CPV (98.15%). Phylogenetic analysis indicated that the isolated FPV formed a monophyletic branch in FPV cluster which differed from the other 11 FPV isolates from China and other countries. The isolatedvirus caused typical clinical symptoms of FPV in cats. This is the first report on isolation of FPV from a monkey. PMID:20044220
SUMMARY Twelve influenza A viruses, antigenically related to the Ho, HI and Hswl subtypes, were isolated from cloacal samples of feral ducks in Canada. Antigenic comparisons showed that these viruses were most closely related to the recent HswINI isolates from man and pigs, whereas in vivo pathogenicity tests revealed differences between the HswINl viruses from the ducks and those from
Avian influenza A viruses are the ancestral origin of all human influenza viruses. The outbreak of highly pathogenic (HP) avian H5N1 in Hong Kong in 1997 highlighted the potential of these viruses to infect and cause severe disease in humans. Since 1999, HP H5N1 viruses were isolated several times from domestic poultry in Asia. In 2001, a HP H5N1 virus, A/Duck/Anyang/AVL-1/2001 (Dk/Anyang), was isolated from imported frozen duck meat in Korea. Because of this novel source of HP H5N1 virusisolation, concerns were raised about the potential for human exposure and infection; we therefore compared the Dk/Anyang virus with HP H5N1 virusesisolated from humans in 1997 in terms of antigenicity and pathogenicity for mammals. At high doses, Dk/Anyang virus caused up to 50% mortality in BALB/c mice, was isolated from the brains and lymphoid organs of mice, and caused lymphopenia. Overall Dk/Anyang virus was substantially less pathogenic for mice than the H5N1 virusisolated from a fatal human case in 1997. Likewise, Dk/Anyang virus was apathogenic for ferrets. Dk/Anyang virus was antigenically distinguishable by hemagglutination-inhibition (HI) assay from human H5N1 virusesisolated in 1997 and avian H5N1 virusesisolated in 2001 in Hong Kong. Nevertheless, prior infection with Dk/Anyang virus protected mice from death after secondary infection with HP human H5N1 viruses. These results indicate that compared with HP human H5N1 viruses, Dk/Anyang virus is substantially less pathogenic for mammalian species. Nevertheless, the novel source of isolation of this avian H5N1 virus must be considered when evaluating the potential risk to public health. PMID:12601764
During the last bovine vaccinia (BV) outbreaks, several Vaccinia virus (VACV) strains were isolated and characterised, revealing significant polymorphisms between strains, even within conserved\\u000a genes. Although the epidemiology of VACV has been studied in BV outbreaks, there is little data about the circulation of the\\u000a Brazilian VACV isolates. This study describes the genetic and biological characterisation of two VACV isolates,
Rafael K. Campos; Mário C. S. Brum; Carlos E. W. Nogueira; Betânia P. Drumond; Pedro A. Alves; Larissa Siqueira-Lima; Felipe L. Assis; Giliane S. Trindade; Cláudio A. Bonjardim; Paulo C. Ferreira; Rudi Weiblen; Eduardo F. Flores; Erna G. Kroon; Jônatas S. Abrahăo
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
Thirty-three stool specimens from 29 patients were examined for Coxsackie virus by the inoculation of suckling mice. Such a virus, designated “California I,” was obtained from two stool specimens collected on successive days from a patient with so-called nonparalytic poliomyelitis. Neutralizing antibodies for the California I strain of Coxsackie virus could not be demonstrated in serum obtained from the patient early in the illness, but were present in convalescent serum. Serum from the patient's daughter, who previously had had a similar illness, neutralized the strain of virusisolated from the father. In pathologic examination of the skeletal muscles of mice infected with the California I virus, lesions typical of those produced by Coxsackie virus, group A, were noted. California I strain of the virus was not neutralized by immune serum prepared from several other strains of Coxsackie virus.
Boak, Ruth A.; Klaumann, Benjamin F.; Ward, C. Bradley
This report describes the genetic and antigenic variability of bovine viral diarrhea virus strains isolated in Belgium. Part of the 5? untranslated region and the 5? end of the gp53 (E2) coding sequence were amplified by PCR and sequenced. Phylogenetic analysis showed that most field isolates segregated into genotypes Ib or II. Only one out of 28 field isolates belonged
B. Couvreur; C. Letellier; A. Collard; P. Quenon; P. Dehan; C. Hamers; P.-P. Pastoret; P. Kerkhofs
Genetic characterization of a human cerebrospinal fluid West Nile virusisolate from Beaumont, Texas, revealed several nucleotide changes and amino acid substitutions that differentiated it from all other North American strains isolated to date, suggesting that isolates from the Texas Gulf Coast may form a unique genetic group among North American strains. PMID:15528747
Granwehr, Bruno P; Li, Li; Davis, Charles T; Beasley, David W C; Barrett, Alan D T
SUMMARY Infective clones of the Nigerian isolate of cassava latent virus (CLV) have been obtained. The apparent molecular weight of the capsid protein of this isolate is slightly higher than that produced in plants infected with cloned DNAs of the Kenyan isolate of CLV. Pseudorecombinant experiments using heterologous combinations of cloned DNAs have confirmed that the physical properties of the
The viral determinants that underlie human immunodeficiency virus type 1 (HIV-1) neurotropism are unknown, due in part to limited studies on virusesisolated from brain. Previous studies suggest that brain- derived viruses are macrophage tropic (M-tropic) and principally use CCR5 for virus entry. To better understand HIV-1 neurotropism, we isolated primary viruses from autopsy brain, cerebral spinal fluid, blood, spleen,
PAUL R. GORRY; GREG BRISTOL; JEROME A. ZACK; KIMBERLY RITOLA; RONALD SWANSTROM; CHRIS J. BIRCH; JEANNE E. BELL; NORBERT BANNERT; KEITH CRAWFORD; HUI WANG; DOMINIQUE SCHOLS; ERIK DE CLERCQ; KEVIN KUNSTMAN; STEVEN M. WOLINSKY; DANA GABUZDA
In late 1998, Nipah virus emerged in peninsular Malaysia and caused fatal disease in domestic pigs and humans and substantial economic loss to the local pig industry. Surveillance of wildlife species during the outbreak showed neutralizing antibodies to Nipah virus mainly in Island flying-foxes (Pteropus hypomelanus) and Malayan flying-foxes (Pteropus vampyrus) but no virus reactive with anti-Nipah virus antibodies was
Kaw Bing Chua; Chong Lek Koh; Poh Sim Hooi; Kong Fatt Wee; Jenn Hui Khong; Beng Hooi Chua; Yee Peng Chan; Mou Eng Lim; Sai Kit Lam
During October-November 1990, 31,497 mosquitoes consisting of 25 different species were collected in Barkedji, Ferlo area (Senegal), and tested for virus infection. Viruse were isolated from 55 of 407 pools. Eighteen pools were found positive for both Bagaza virus (BGA) and West Nile virus (WN). One alphavirus (Babanki [BBK] and 72 flaviviruses (19 BGA, 53 WN) were isolated from Culex poicilipes Theobald (29 WN, 8 BGA), C. neavei Theobald (3 WN, 1 BGA), Mimomyia hispida Theobald (8 WN, 6 BGA, and 1 BBK), M. lacustris Edwards (4 WN, 1 BGA), M. splendens Theobald (6 WN, 2 BGA), Mimomyia. spp. (2 WN), and Aedeomyia africana Neveu-Lemaire (1 WN). These were the first isolations of arboviruses from A. africana and Mimomyia species. C. poicilipes and possibly Mimomyia spp. may be involved in an avian-mosquito cycle of West Nile virus transmission in Senegal. PMID:7815413
Traore-Lamizana, M; Zeller, H G; Mondo, M; Hervy, J P; Adam, F; Digoutte, J P
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
Brian Wigdahl; Carol A. Smith; Helen M. Traglia; Fred Rapp
Virusesisolated from the blood of two Korean haemorrhagic fever patients were propagated in cell culture and compared to prototype Hantaan virus which was isolated from Apodemus mice. The antigenic properties of the human isolates were found to be closely related to Hantaan virus by plaque reduction neutralization, haemagglutination inhibition and fluorescent antibody staining with both polyclonal and monoclonal antibodies. The medium genome segment of each human isolate was sequenced and compared to that of Hantaan virus. Nucleotides comprising the Hantaan virus G1 and G2 envelope protein-coding regions differed from those of the other viruses by only 5.4% and 5.7%. The human isolates differed from one another by 1.6%. The nucleotide differences resulted in predicted amino acid variations of 1.3% to 2.3% among the three viruses, with the majority occurring as conservative substitutions in G1. PMID:2900289
Schmaljohn, C S; Arikawa, J; Hasty, S E; Rasmussen, L; Lee, H W; Lee, P W; Dalrymple, J M
We have previously described the development of AIDS in a chimpanzee (C499) infected with human immunodeficiency virus type 1 (HIV-1) and the subsequent pathogenic HIV-1 infection in another chimpanzee (C455) transfused with blood from C499 (F. J. Novembre et al., J. Virol. 71:4086–4091, 1997). In the present study, two virusisolates were derived from these animals: HIV-1JC from peripheral blood mononuclear cells (PBMC) of C499, and HIV-1NC from plasma of C455. These virusisolates were used to generate two infectious molecular clones, termed HIV-1JC16 and HIV-1NC7 (JC16 and NC7, respectively). Comparative analyses of the sequences of the two clones showed that they were highly interrelated but distinct. Based on heteroduplex mobility assays, JC16 and NC7 appear to represent dominant viruses in the uncloned stock population. Compared with amino acid sequences of the parental viruses HIV-1SF2, HIV-1LAV-1b, and HIV-1NDK, JC16 and NC7 showed a number of differences, including insertions, deletions, and point mutations spread throughout the genome. However, insertion/deletion footprints in several genes of both JC16 and NC7 suggested that recombination between SF2 and LAV-1b could have occurred, possibly contributing to the generation of a pathogenic virus. Comparative in vitro analyses of the molecular clones and the uncloned stocks of HIV-1JC and HIV-1NC revealed that these viruses had strikingly similar replicative abilities in mitogen-stimulated PBMC and in macrophages. Compared to the SF2 and LAV-1b isolates of HIV-1, HIV-1JC and HIV-1NC isolates were more similar to LAV-1b with respect to the ability to replicate in mitogen-stimulated PBMC and macrophages. These viruses should prove to be useful in mapping determinants of pathogenesis.
We have previously described the development of AIDS in a chimpanzee (C499) infected with human immunodeficiency virus type 1 (HIV-1) and the subsequent pathogenic HIV-1 infection in another chimpanzee (C455) transfused with blood from C499 (F. J. Novembre et al., J. Virol. 71:4086-4091, 1997). In the present study, two virusisolates were derived from these animals: HIV-1JC from peripheral blood
Following the reemergence of Rift Valley fever (RVF) virus in southeastern Mauritania in 1998, an entomological survey was undertaken in the boundary area in Senegal to assess the extent of the virus circulation. During this study, RVF virus (36 strains) was isolated for the first time from Culex poicilipes in nature. The possible role of Cx. poicilipes as an RVF vector is discussed regarding its biology and ecology. PMID:11304058
Diallo, M; Lochouarn, L; Ba, K; Sall, A A; Mondo, M; Girault, L; Mathiot, C
From biopsies taken from the vaginal tract of dairy cattle a virus was isolated in embryonated eggs. This virus was cytopathogenic to chick kidney and bovine embryo cell cultures with the formation of plaques on the former. Antisera for Infectious Pustular Vulvovaginitis, Enteric Cytopathogenic Bovine Orphan, Chick Embryo Lethal Orphan, Newcastle disease, Infectious Bronchitis, and Laryngotracheitis failed to neutralize the virus. ImagesFigure 1.Figure 2.Figure 3.Figure 4.
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
T. S. Gritsun; T. V. Frolova; A. I. Zhankov; M. Armesto; S. L. Turner; M. P. Frolova; V. V. Pogodina; V. A. Lashkevich; E. A. Gould
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.
The complete sequence of RNA 3 of the Blencowe, British (B) isolate of tomato aspermy virus (TAV) is presented. The RNA 3 of TAV-B is dicistronic, encoding the putative movement protein 3a and the capsid protein.
David O'Reilly; Chris J. R. Thomas; Robert H. A. Coutts
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.
Souza, Victor C.; Silva, George A. V.; Maito, Rodrigo M.; Granja, Fabiana; Siqueira, Thalita; Acosta, Pablo O. A.
Dengue is the most important arboviral disease worldwide. We report the complete genome sequence of a dengue virus serotype 4, genotype II strain isolated in 2010 from a patient with classical dengue fever in Boa Vista, Roraima, Brazil. PMID:22247521
Naveca, Felipe G; Souza, Victor C; Silva, George A V; Maito, Rodrigo M; Granja, Fabiana; Siqueira, Thalita; Acosta, Pablo O A
Summary 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.
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
Four H7N3 avian influenza viruses (AIVs) were isolated from domestic ducks in live-poultry markets in Zhejiang Province, Eastern China, in 2011. All viruses were characterized by whole-genome sequencing with subsequent phylogenetic analysis and genetic comparison. Phylogenetic analysis of all eight viral genes showed that the viruses clustered in the Eurasian lineage of influenza viruses. The hemagglutinin cleavage site of all viruses indicated that the four strains were low-pathogenic avian influenza viruses. PMID:22752840
A circular, viroid-like satellite RNA (sat RNA) was detected in lucerne transient streak virus (LTSV), from lucerne in South Australia. It was larger than the previously reported sat RNA of LTSV, being similar in size and sequence homology to the 388 nucleotide sat RNA previously shown to be encapsidated by subterranean clover mottle virus (SCMoV) isolated from subterranean clover in Western Australia. This indicates that under field conditions, very similar sat RNAs can be associated with two distantly related sobemoviruses, LTSV and SCMoV. The natural hosts of these viruses are lucerne and subterranean clover, respectively. PMID:1697331
Dall, D J; Graddon, D J; Randles, J W; Francki, R I
In studies designed to investigate the role of snakes in the ecology of Japanese encephalitis (JE) virus in Korea, 747 snakes representing 4 different species were collected during 1966 and 1967. Two strains of JE virus were isolated from the snakes, E. r...
sequence data on H9 avian influenza virus (AIV) from wild birds are currently limited, we set out to determine the sequence of the hemagglutinin (HA) gene of H9 viruses circulating in North American waterfowl and shorebirds. In this study, we examined the HA gene from H9 AIV isolated from mallards (Anas platyrhynchos) sampled during 1998 and 1999 in Minnesota and
Dengue virus was detected for the first time in Chile, in an outbreak of dengue fever on Easter Island. The virus was isolated in tissue culture and characterized by reverse transcription-polymerase chain reaction as being dengue type 1. PMID:14718094
Wild birds are considered to be the natural reservoirs for avian influenza A viruses (AIV). During active influenza surveillance in Poyang Lake of southeast China, we isolated and characterized 11 H9N2 viruses from two species of wild ducks. Phylogenetic analysis showed that the 11 isolates were almost identical with 99.3-100% nucleotide homology in their entire genome, and they all closely related in whole eight genes (95.6-99.4% homology) to human H9N2 isolates (HK/33982/2009) and clustered in the same sublineage. The isolates belonged to triple reassortant H9N2 genotype viruses containing Ck/Bei-like NA genes, Y439-like PA genes and six other G1-like genes. We also found that the subtype of virus replicated efficiently in the lungs and tracheas of BALB/c mice and caused mortality in 20-40% of infected groups after 3-6 days, which indicates that the subtype of virus is capable of establishing lethal mammalian infections. However, whether or not the virus has features transmittable from wild ducks to humans is not known. This study showed H9N2 subtype avian influenza virus for the first time in wild birds, and suggests that wild birds may carry the virus along migratory routes, highlighting the need for continued surveillance of wild birds. PMID:23830774
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
Jean-Louis Zeddam; Juana Luna Rodriguez; Marc Ravallec; Aziz Lagnaoui
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 ...
Summary Previous serological studies strongly suggested Akabane virus to be the etiologic agent of epizootic abortion and congenital arthrogryposis-hydranencephaly in cattle, and this view was further corroborated in this study by the isolation of the virus from an aborted fetus in an epizootic of the disease and from a fetus extracted from a cow which was suggested by serologic tests
H. Kurogi; Y. Inaba; E. Takahashi; K. Sato; T. Omori; Y. Miura; Y. Goto; Y. Fujiwara; Y. Hatano; K. Kodama; S. Fukuyama; N. Sasaki; M. Matumoto
Biological and biochemical properties of four nuclear polyhedrosis virusisolates from beet armyworm, Spodoptera exigua, were investigated. The isolates originated from the United States (SeNPV?US), Thailand (SeNPV?TH) and from two locations in Spain (SeNPV?SP1 and SeNPV?SP2). Restriction endonuclease analysis of the viral genomes revealed limited restriction fragment length polymorphism and indicated that these viruses contained distinct, but closely related, genotypes
Dengue virus infection has been recognized as an important public health problem in the Dominican Republic in the last decade. Complete genomic sequences of three strains of dengue type 2 (DEN-2) virus, DR23\\/01 and DR31\\/01 isolated from dengue fever (DF) patients, and DR59\\/01 isolated from a dengue hemorrhagic fever (DHF) patient, all with primary infection, in the Dominican Republic in
A feline panleukopenia virus (FPV) mutant, monkey\\/BJ-22\\/2008\\/CHN, was isolated from intestinal contents of a diarrheic monkey in Beijing, China. The virus was identified by morphology and physicochemical characteristics, and specific fragments were obtained by PCR using consensus primers of parvovirus and specific primers of FPV. Sequence of the full-length VP2 gene of the isolated FPV was determined and analyzed by
Songtao Yang; Shujun Wang; Hao Feng; Lin Zeng; Zhiping Xia; Renzhou Zhang; Xiaohuan Zou; Chengyu Wang; Quan Liu; Xianzhu Xia
A rabies virus (RABV) was isolated from a dog in Anhui Province, China, in 2008. The virus was designated DRV-AH08. Its entire genome was sequenced and found to be closely related to RABV recently isolated in China and other Asian countries (homology of 87 to 98%) but distantly related to RABV in the "cosmopolitan" group (homology of 84 to 85%) in the clade I of RABV. PMID:22966185
A rabies virus (RABV) was isolated from a dog in Anhui Province, China, in 2008. The virus was designated DRV-AH08. Its entire genome was sequenced and found to be closely related to RABV recently isolated in China and other Asian countries (homology of 87 to 98%) but distantly related to RABV in the “cosmopolitan” group (homology of 84 to 85%) in the clade I of RABV.
Isolates of Cucumber green mottle mosaic virus (CGMMV) obtained from three greenhouses of Ukraine were identified. The rabbit antisera have been obtained for these isolates. These antisera can be used for diagnostics of CGMMV in plant material by indirect ELISA. An analysis of cDNA sequences of capsid protein of these viruses confirmed that they might be grouped into the novel strain of CGMMV. PMID:16493891
Rudnieva, T O; Budzanivs'ka, I H; Ryzhkova, A Ie; Shevchenko, T P; Dem'ianenko, F P; Polishchuk, V P
Dasheen mosaic virus (DsMV) is an important constraint to production of cocoyam (Xanthosoma spp.) in Nicaragua. Reverse transcription polymerase chain reaction was used to amplify the coat protein (CP) region from\\u000a ten Nicaraguan DsMV isolates. These isolates showed high nucleotide identity to DsMV isolates from the USA, eastern Asia and\\u000a Australasia. All Nicaraguan isolates except one shared a tandem repeat
Guillermo Reyes; Jon N. E. Ramsell; Marie Nyman; Anders Kvarnheden
An isolate of Beet black scorch virus (BBSV) was obtained from Iranian sugar beet roots. Its genome organization closely resembles that of the previously described\\u000a Chinese and North American isolates, but the nucleotide sequences of the three isolates differ considerably. Most of the nucleotide\\u000a exchanges, however, are silent, and the Iranian and the Chinese isolates were serologically indistinguishable. Beets infected
In this study, we isolated and characterized an insect nidovirus from the mosquito Culex tritaeniorhynchus Giles (Diptera: Culicidae) in Vietnam, as an additional member of the new family Mesoniviridae in the order Nidovirales. The virus, designated "Dak Nong virus (DKNV)," shared many characteristics with Cavally virus and Nam Dinh virus, which have also been discovered recently in mosquitoes, and these viruses should be considered members of a single virus species, Alphamesonivirus 1. DKNV grew in cultured mosquito cells but could not replicate in the cultured vertebrate cells tested. N-terminal sequencing of the DKNV structural proteins revealed two posttranslational cleavage sites in the spike glycoprotein precursor. DKNV is assumed to be a new member of the species Alphamesonivirus 1, and the current study provides further understanding of viruses belonging to the new family Mesoniviridae. PMID:23728735
Thirteen oncogenic and necrotizing animal viruses were assayed in LIFE Sciences, Inc. (LSI)-specific pathogen-free Japanese quail and LSI-specific pathogen-free chicken embryo cell cultures. Nine viruses produced similar titers in the quail and chicken cell systems, whereas four viruses showed significantly higher titers in chickens. Young Japanese quail and chickens were inoculated with five selected avain viruses and maintained in stainless-steel isolators. Comparable responses were noted in quail and chickens injected with Newcastle disease virus and avain leukosis virus, but quail were significantly more resistant than chickens to fowl pox virus, laryngotracheitis virus, and Marek's disease herpesvirus. Although no overt symptoms of disease were observed in Japanese quail inoculated with most avain viruses, neutralizing antibody or virus was detected, indicating presence of an inapparent infection. In one experiment, neutralizing antibody was detected in a comparable number of quail and chickens after inoculation with avian leukosis virus. Avian leukosis virus viremia was observed at 12 and 70 days postinoculation, with the COFAL (complement fixation for avian leukosis) titers similar for quail and chickens. Most quail infected with Marek's disease herpesvirus produced neutralizing antibody within 70 days but showed no classical symptoms of Marek's disease even when held for 5 months. In contrast, all chickens inoculated with Marek's disease herpesvirus died within 20 days. The utility of quail embryo cell cultured in the preparation of vaccines and biological reagents is discussed.
Embryo lethality patterns induced by an avian influenza virusisolate (A/Tk/Ws/68/H5N9) suggested that it contained more than one genetic form. Two different virus populations were recovered from the isolate by plaque isolation and limit-dilution cloning and were characterized with respect to their biological and molecular properties. They were very closely related but exhibited strikingly different mean death times (MDT) in 10-day-old chick embryos. One was rapidly embryo lethal (REL), while the other was slowly embryo lethal (SEL). The REL isolate demonstrated a small but measurable mortality rate in 4-week-old chicks, as did the parental isolate. The SEL isolate, however, was nonlethal to 4-week-old chicks. The embryo MDT induced by the parental isolate revealed a biphasic death pattern reflecting expression of both REL and SEL populations. Mixing experiments, using different amounts of the two cloned populations, demonstrated that expression of their unique phenotypic property (either REL or SEL) was competitive. The number of early or late embryo deaths was directly related to the input levels of each respective virus. The only molecular difference thus far detected between the two populations is in the nonstructural (NS) gene, with the REL clone possessing a faster migrating electrophoretic form of that RNA than the SEL clone. Both forms of the NS gene were present in the original parental isolate. This study thus demonstrates the competitive coexistence of two closely related virus populations within a single natural isolate. PMID:2619663
The rarely identified influenza A viruses of the H15 hemagglutinin subtype have been isolated exclusively in Australia. Here we report the isolation of an H15N4 influenza A virus (A/teal/Chany/7119/2008) in Western Siberia, Russia. Phylogenetic analysis demonstrated that the internal genes of the A/teal/Chany/7119/2008 strain belong to the Eurasian clade and that the H15 and N4 genes were introduced into the gene pool of circulating endemic avian influenza viruses through reassortment events.
Sivay, Mariya V.; Baranovich, Tatiana; Marchenko, Vasiliy Y.; Sharshov, Kirill A.; Govorkova, Elena A.; Shestopalov, Aleksander M.
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.
Palacios, Gustavo; Savji, Nazir; Hui, Jeffrey; Travassos da Rosa, Amelia; Popov, Vsevolod; Briese, Thomas; Tesh, Robert; Lipkin, W. Ian
An outbreak of encephalitis due to West Nile (WN) virus occurred in New York City and the surrounding areas during 1999. Mosquitoes were collected as part of a comprehensive surveillance program implemented to monitor the outbreak. More than 32,000 mosquitoes representing 24 species were tested, and 15 WN virusisolates were obtained. Molecular techniques were used to identify the species represented in the WN virus-positive mosquito pools. Most isolates were from pools containing Culex pipiens mosquitoes, but several pools contained two or more Culex species.
Nasci, R. S.; White, D. J.; Stirling, H.; Oliver, J. A.; Daniels, T. J.; Falco, R. C.; Campbell, S.; Crans, W. J.; Savage, H. M.; Lanciotti, R. S.; Moore, C. G.; Godsey, M. S.; Gottfried, K. L.; Mitchell, C. J.
The rarely identified influenza A viruses of the H15 hemagglutinin subtype have been isolated exclusively in Australia. Here we report the isolation of an H15N4 influenza A virus (A/teal/Chany/7119/2008) in Western Siberia, Russia. Phylogenetic analysis demonstrated that the internal genes of the A/teal/Chany/7119/2008 strain belong to the Eurasian clade and that the H15 and N4 genes were introduced into the gene pool of circulating endemic avian influenza viruses through reassortment events. PMID:23283950
Sivay, Mariya V; Baranovich, Tatiana; Marchenko, Vasiliy Y; Sharshov, Kirill A; Govorkova, Elena A; Shestopalov, Aleksander M; Webby, Richard J
In 2009, 2589 mosquitoes were collected in northwest Italy and screened for orthobunyavirus RNA by RT-PCR. One pool of Anopheles maculipennis complex mosquitoes was found to be positive and a virus was isolated from that pool. The isolate was identified as Batai virus (BATV) by sequencing. Previously, BATV was detected in Italy, but limited data and no prior isolates existed. Full-length sequences of the S, M and L segments were determined for the newly isolated Italian strain. For comparison, partial sequences were also determined for the BATV strain Calovo (former Czechoslovakia, 1960). Phylogenetic analyses revealed clustering of the newly derived Italian BATV along with a recent isolate from Germany and the historic strain Calovo. To the best of our knowledge, this represents the first isolation of BATV from Italy, which confirms a broader geographical distribution of BATV in Europe than was previously verified by isolation. PMID:23515020
Huhtamo, Eili; Lambert, Amy J; Costantino, Stefano; Servino, Luca; Krizmancic, Letizia; Boldorini, Renzo; Allegrini, Sara; Grasso, Ivan; Korhonen, Essi M; Vapalahti, Olli; Lanciotti, Robert S; Ravanini, Paolo
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
Two new rhabdoviruses, designated Carajas and Maraba, are described. Both were isolated from phlebotomine sand flies (Lutzomyia spp.) collected in the Amazon basin of Brazil. One recovery of Carajas virus was made from male sand flies. By complement-fixation and neutralization tests both agents were shown to be members of the vesicular stomatitis virus (VSV) serogroup (genus Vesiculovirus). The pathogenicity of the two viruses in mice and Vero cells is similar to that of VSV-Indiana and VSV-New Jersey. Both Carajas and Maraba viruses replicated in Lutzomyia longipalpis following intrathoracic inoculation, and both agents were transovarially transmitted in this sand fly species. PMID:6091472
Travassos da Rosa, A P; Tesh, R B; Travassos da Rosa, J F; Herve, J P; Main, A J
Background Three influenza pandemics outbroke in the last century accompanied the viral antigen shift and drift, resulting in the change of antigenic property and the low cross protective ability of the existed antibody to the newly emerged pandemic virus, and eventually the death of millions of people. The antigenic characterizations of the virusesisolated in central China in 2004 and 2006–2007 were investigated in the present study. Results Hemagglutinin inhibition assay and neutralization assay displayed differential antigenic characteristics of the virusesisolated in central China in two periods (2004 and 2006–2007). HA genes of the viruses mainly located in two branches in phylogeny analysis. 53 mutations of the deduced amino acids of the HA genes were divided into 4 patterns. Mutations in pattern 2 and 3 showed the main difference between virusesisolated in 2004 and 2006–2007. Meanwhile, most amino acids in pattern 2 and 3 located in the globular head of the HA protein, and some of the mutations evenly distributed at the epitope sites. Conclusions The study demonstrated that a major antigenic drift had occurred in the virusesisolated in central China. And monitoring the antigenic property should be the priority in preventing the potential pandemic of H5N1 avian influenza virus.
Peste des petits ruminants virus is a member of Morbillivirus Paramyxoviridae. The complete genome of a Peste des petits ruminants virus (PPRV) isolate, China/Tib/07 was sequenced and molecular characteristics was analyzed. The internal sequences of the virus genome were amplified by RT-PCR with primers designed according to the published data in GenBank, while the sequences of the 3' and 5' ends of the genome were determined by RACE. Amplification products were directly sequenced,assembled and analyzed with DNAStar4.0. Results showed that China/Tib/07 genome consisted of 15 948 nucleotides in length, encoding six structural proteins and two non-structural proteins just like other known PPRV genomes. Phylogenetically, the virus genome shared homology of 91.6%-98.1% with Southwest Asian isolates among PPRV strains and the highest homology of 64.3% with rinderpest virus among morbillivirus members. PMID:20836387
During 2004-2006 swine influenza virus surveillance, two strains of H3N8 influenza viruses were isolated from pigs in central China. Sequence and phylogenetic analyses of eight gene segments revealed that the two swine isolates were of equine origin and most closely related to European equine H3N8 influenza viruses from the early 1990s. Comparison of hemagglutinin (HA) amino acid sequences showed several important substitutions. One substitution caused the loss of a potential glycosylation site, and two substitutions, located at the cleavage site and adjacent to the receptor-binding pocket, respectively, had been reported previously in canine H3 HAs. This expansion of host range of equine H3N8 influenza viruses with mutations in the HA protein might raise the possibility of transmission of these viruses to humans. PMID:19396578
Thirteen orf virusisolates obtained during the time period between 1995 and 2004 from crusted scab lesions of nine sheep and four goats from different geographical areas of Greece and Italy with suspected contagious ecthyma infection were analyzed. DNA of all isolates was successfully amplified by PCR with the primers 045F-045R and identified them as parapox virus. Partial DNA sequence of orf virus interferon resistant (VIR) gene, phylogenetic analysis of the available isolates and amino acid comparison of the interferon resistance protein encoded by this genomic region was carried out. According to the results of the present report a precise characterisation of the genomic region studied might provide evidence for the genetic variation and movement of the circulating orf virus strains. PMID:16757136
Jamestown Canyon (JC) virus of the California (CAL) serogroup has been isolated in 12 American states and 6 Canadian provinces. A study was undertaken to produce monoclonal antibodies (MAbs) to JC virus and to use these MAbs to assay for possible heterogeneity among naturally occurring JC topotypes in Canada. MAbs were produced to the prototype strain of JC virus using BALB/c mice. Twenty-seven secreting MAbs were obtained and three of these MAbs were propagated and studied. All three MAbs, M1 (IgG1), M2 (IgG2b), and M3 (IgG2a), were reactive by immunofluorescent antibody assay against JC-infected vero cells and by ELISA against JC antigen. MAb M2 reacted with all members of the Melao complex, MAb M1 reacted only with Keystone virus, while MAb M3 exhibited no reactivity with other CAL serogroup viruses. Only MAb M3 possessed neutralization and hemagglutination inhibition activities against JC virus. The MAbs were also tested by ELISA and for neutralizing activity against 13 JC topotypes isolated in 5 provinces from Newfoundland to Saskatchewan. ELISA confirmed closer identity of the Canadian topotypes to JC as opposed to the closely related South River virus. The MAbs verified all Canadian topotypes to be JC virus but revealed different patterns of reactivity between these topotypes and prototype JC virus. PMID:1418321
The sooty mangabey-derived simian immunodeficiency virus (SIV) strain E660 (SIVsmE660) is a genetically heterogeneous, pathogenic isolate that is commonly used as a vaccine challenge strain in the nonhuman primate (NHP) model of human immunodeficiency virus type 1 (HIV-1) infection. Though it is often employed to assess antibody-based vaccine strategies, its sensitivity to antibody-mediated neutralization has not been well characterized. Here, we utilize single-genome sequencing and infectivity assays to analyze the neutralization sensitivity of the uncloned SIVsmE660 isolate, individual viruses comprising the isolate, and transmitted/founder (T/F) viruses arising from low-dose mucosal inoculation of macaques with the isolate. We found that the SIVsmE660 isolate overall was highly sensitive to neutralization by SIV-infected macaque plasma samples (50% inhibitory concentration [IC50] < 10?5) and monoclonal antibodies targeting V3 (IC50 < 0.01 ?g/ml), CD4-induced (IC50 < 0.1 ?g/ml), CD4 binding site (IC50 ? 1 ?g/ml), and V4 (IC50, ?5 ?g/ml) epitopes. In comparison, SIVmac251 and SIVmac239 were highly resistant to neutralization by these same antibodies. Differences in neutralization sensitivity between SIVsmE660 and SIVmac251/239 were not dependent on the cell type in which virus was produced or tested. These findings indicate that in comparison to SIVmac251/239 and primary HIV-1 viruses, SIVsmE660 generally exhibits substantially less masking of antigenically conserved Env epitopes. Interestingly, we identified a minor population of viruses (?10%) in both the SIVsmE660 isolate and T/F viruses arising from it that were substantially more resistant (>1,000-fold) to antibody neutralization and another fraction (?20%) that was intermediate in neutralization resistance. These findings may explain the variable natural history and variable protection afforded by heterologous Env-based vaccines in rhesus macaques challenged by high-dose versus low-dose SIVsmE660 inoculation regimens.
Lopker, Michael; Easlick, Juliet; Sterrett, Sarah; Decker, Julie M.; Barbian, Hannah; Learn, Gerald; Keele, Brandon F.; Robinson, James E.; Li, Hui; Hahn, Beatrice H.; Shaw, George M.
The isolation of infectious salmon anaemia virus (ISAV) from asymptomatic wild fish species including wild salmon, sea trout and eel established that wild fish can be a reservoir of ISAV for farmed Atlantic salmon. This report characterizes the biological properties of ISAV isolated from a disease outbreak in farmed Coho salmon in Chile and compares it with ISAV isolated from farmed Atlantic salmon in Canada and Europe. The virus that was isolated from Coho salmon tissues was initially detected with ISAV-specific RT-PCR (reverse transcription-polymerase chain reaction). The ability of the virus to grow in cell culture was poor, as cytopathology was not always conspicuous and isolation required passage in the presence of trypsin. Virus replication in cell culture was detected by RT-PCR and IFAT (indirect fluorescent antibody test), and the virus morphology was confirmed by positive staining electron microscopy. Further analysis of the Chilean virus revealed similarities to Canadian ISAV isolates in their ability to grow in the CHSE-214 cell line and in viral protein profile. Sequence analysis of genome segment 2, which encodes the viral RNA polymerase PB1, and segment 8, which encodes the nonstructural proteins NS1 and NS2, showed the Chilean virus to be very similar to Canadian strains of ISAV. This high sequence similarity of ISAV strains of geographically distinct origins illustrates the highly conserved nature of ISAV proteins PB1, NS1 and NS2 of ISAV. It is noteworthy that ISAV was associated with disease outbreaks in farmed Coho salmon in Chile without corresponding clinical disease in farmed Atlantic salmon. This outbreak, which produced high mortality in Coho salmon due to ISAV, is unique and may represent the introduction of the virus to a native wild fish population or a new strain of ISAV. PMID:11411649
Kibenge, F S; Gárate, O N; Johnson, G; Arriagada, R; Kibenge, M J; Wadowska, D
On April 21, 2008, four whooper swans were found dead at Lake Towada, Akita prefecture, Japan. Highly pathogenic avian influenza virus of the H5N1 subtype was isolated from specimens of the affected birds. The hemagglutinin (HA) gene of the isolate belongs to clade 2.3.2 in the HA phylogenetic tree.
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...
Swine influenza virusisolates originating from outbreaks in Sweden from 1983, 2002 and 2009 were subjected to nucleotide sequencing and phylogenetic analysis. The aim of the studies was to obtain an overview on their potential relatedness as well as to provide data for broader scale studies on swine influenza epidemiology. Nonetheless, analyzing archive isolates is justified by the efforts directed
István Kiss; Ádám Bálint; Giorgi Metreveli; Eva Emmoth; Frederik Widén; Sándor Belák; Per Wallgren
Isolates of Banana bunchy top virus (BBTV), which causes bunchy top disease in bananas, were collected in field surveys on seven islands in Okinawa Prefecture, Japan. From 44 banana samples, one isolate from each island was selected, and the DNA-1 and DNA-3 components were sequenced. Analysis of the major common region of DNA-1 showed that BBTV in Okinawa belongs to
WUH4 is a highly pathogenic North American porcine reproductive and respiratory syndrome virus (PRRSV). Unlike previous PRRSV isolates, which were mainly recovered from sera or tissues, WUH4 was isolated from a piglet stool sample. Here we announce its complete genome sequence.
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.
Porcine epidemic diarrhea virus (PEDV) has plagued the domestic swine industry in Korea causing significant economic impacts on pig production nationwide. In the present study, we determined the complete nucleotide sequences of the spike (S) glycoprotein genes of seven Korean PEDV isolates. The entire S genes of all isolates were found to be nine nucleotides longer in length than other
Dong-Kyu Lee; Choi-Kyu Park; Seong-Hee Kim; Changhee Lee
The aim of the study was to select, from a panel of candidate European human immunodeficiency virus type 1 (HIV-1) clade B primary virusisolates, one isolate based on replication properties in chim- panzee peripheral blood mononuclear cells (PBMC). Secondly, to evaluate the in vivo kinetics of primary infection of the selected isolate at two different doses in two mature,
W. M. J. M. Bogers; W. H. Koornstra; R. H. Dubbes; Haaft ten P. J. F; B. E. Verstrepen; S. S. Jhagjhoorsingh; A. G. M. Haaksma; H. Niphuis; J. D. Laman; S. Norley; H. Schuitemaker; J. Goudsmit; G. Hunsmann; J. L. Heeney; H. Wigzell
The isolation of a number of strains of infectious bursal disease (IBD) virus from fowl, turkeys and ducks is described. These isolates could be grouped into two serotypes using the neutralisation test. It is proposed that the cell culture adapted vaccine strain from fowl should be the prototype virus for serotype 1 and that the TY89 isolate from a turkey
J. B. McFerran; M. S. McNulty; E. R. McKillop; T. J. Connor; R. M. McCracken; D. S. Collins; G. M. Allan
Newcastle disease (ND) is one of the most important diseases of poultry, negatively affecting poultry production worldwide. The disease is caused by Newcastle disease virus (NDV) or avian paramyxovirus type 1 (APMV-1), a negative-sense single-stranded RNA virus of the genus Avulavirus, family Paramyxoviridae. Although all NDV isolates characterized to date belong to a single serotype of APMV-1, significant genetic diversity has been described between different NDV isolates. Here we present the complete genome sequence and the clinicopathological characterization of a virulent Newcastle disease virusisolate (NDV-Peru/08) obtained from poultry during an outbreak of ND in Peru in 2008. Phylogenetic reconstruction and analysis of the evolutionary distances between NDV-Peru/08 and other isolates representing established NDV genotypes revealed the existence of large genomic and amino differences that clearly distinguish this isolate from viruses of typical NDV genotypes. Although NDV-Peru/08 is a genetically distinct virus, pathogenesis studies conducted with chickens revealed that NDV-Peru/08 infection results in clinical signs characteristic of velogenic viscerotropic NDV strains. Additionally, vaccination studies have shown that an inactivated NDV-LaSota/46 vaccine conferred full protection from NDV-Peru/08-induced clinical disease and mortality. This represents the first complete characterization of a virulent NDV isolate from South America.
Diel, Diego G.; Susta, Leonardo; Cardenas Garcia, Stivalis; Killian, Mary L.; Brown, Corrie C.; Afonso, Claudio L.
Dengue-1 viruses responsible for the dengue fever outbreak in Easter Island in 2002 were isolated from acute-phase sera of dengue fever patients. In order to analyze the complete genome sequence, we designed primers to amplify contiguous segments across the entire sequence of the viral genome. RT-PCR products obtained were cloned, and complete nucleotide and deduced amino acid sequences were determined. This report constitutes the first complete genetic characterization of a DENV-1 isolate from Chile. Phylogenetic analysis shows that an Easter Island isolate is most closely related to Pacific DENV-1 genotype IV viruses. PMID:18815724
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. Virusisolation 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.
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.
Osorio, Jorge E.; Ciuoderis, Karl A.; Lopera, Juan G.; Piedrahita, Leidy D.; Murphy, Darby; LeVasseur, James; Carrillo, Lina; Ocampo, Martha C.; Hofmeister, Erik
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.
Osorio, Jorge E.; Ciuoderis, Karl A.; Lopera, Juan G.; Piedrahita, Leidy D.; Murphy, Darby; LeVasseur, James; Carrillo, Lina; Ocampo, Martha C.; Hofmeister, Erik
Seven of nine colostrum deprived calves, free from bovine viral diarrhoea virus (BVDV), were vaccinated with a commercially available vaccine containing two inactivated strains of BVDV, an inactivated strain of bovine herpesvirus-1 and modified-live strains of bovine respiratory syncytial virus and para-influenza-3 virus. The two other calves were kept as controls. The virus neutralising (VN) antibodies induced by vaccination were
C. Hamers; E. Di valentin; C. Lecomte; M. Lambot; E. Joris; B. Genicot; P. Pastoret
We tested chemokine receptor subset usage by diverse, well-characterized primary virusesisolated from peripheral blood by monitoring viral replication with CCR1, CCR2b, CCR3, CCR5, and CXCR4 U87MG.CD4 transformed cell lines and STRL33/BONZO/TYMSTR and GPR15/BOB HOS.CD4 transformed cell lines. Primary viruses were isolated from 79 men with confirmed human immunodeficiency virus type 1 (HIV-1) infection from the Chicago component of the Multicenter AIDS Cohort Study at interval time points. Thirty-five additional well-characterized primary viruses representing HIV-1 group M subtypes A, B, C, D, and E and group O and three primary simian immunodeficiency virus (SIV) isolates were also used for these studies. The restricted use of the CCR5 chemokine receptor for viral entry was associated with infection by a virus having a non-syncytium-inducing phenotype and correlated with a reduced rate of disease progression and a prolonged disease-free interval. Conversely, broadening chemokine receptor usage from CCR5 to both CCR5 and CXCR4 was associated with infection by a virus having a syncytium-inducing phenotype and correlated with a faster rate of CD4 T-cell decline and progression of disease. We also observed a greater tendency for infection with a virus having a syncytium-inducing phenotype in men heterozygous for the defective CCR5 Delta32 allele (25%) than in those men homozygous for the wild-type CCR5 allele (6%) (P = 0.03). The propensity for infection with a virus having a syncytium-inducing phenotype provides a partial explanation for the rapid disease progression among some men heterozygous for the defective CCR5 Delta32 allele. Furthermore, we did not identify any primary viruses that used CCR3 as an entry cofactor, despite this CC chemokine receptor being expressed on the cell surface at a level commensurate with or higher than that observed for primary peripheral blood mononuclear cells. Whereas isolates of primary viruses of SIV also used STRL33/BONZO/TYMSTR and GPR15/BOB, no primary isolates of HIV-1 used these particular chemokine receptor-like orphan molecules as entry cofactors, suggesting a limited contribution of these other chemokine receptors to viral evolution. Thus, despite the number of chemokine receptors implicated in viral entry, CCR5 and CXCR4 are likely to be the physiologically relevant chemokine receptors used as entry cofactors in vivo by diverse strains of primary virusesisolated from blood. PMID:9765480
Zhang, L; He, T; Huang, Y; Chen, Z; Guo, Y; Wu, S; Kunstman, K J; Brown, R C; Phair, J P; Neumann, A U; Ho, D D; Wolinsky, S M
Two swine influenza (SI) H1N1 virus was isolated from a pig during a severe outbreak of respiratory disease in south China.\\u000a The two H1N1 influenza viruses were classical SI virus. A\\/swine\\/Guangdong\\/L6\\/09 is classical SI virus of recent years, which\\u000a is of the main SI virus in China. Howere, A\\/swine\\/Guangdong\\/L3\\/09 was closet to A\\/swine\\/Iowa\\/1931, which was the first isolated\\u000a SI virus
In 1997, 18 human infections with H5N1 influenza type A were identified in Hong Kong and six of the patients died. There were concomitant outbreaks of H5N1 infections in poultry. The gene segments of the human H5N1 viruses were derived from avian influenza A viruses and not from circulating human influenza A viruses. In 1999 two cases of human infections caused by avian H9N2 virus were also identified in Hong Kong. These events established that avian influenza viruses can infect humans without passage through an intermediate host and without acquiring gene segments from human influenza viruses. The likely origin of the H5N1 viruses has been deduced from molecular analysis of these and other virusesisolated from the region. The gene sequences of the H5N1 viruses were analysed in order to identify the molecular basis for the ability of these avian viruses to infect humans. PMID:11015744
SUMMARY The neuraminidases and haemagglutinins of 20 isolates of swine influenza virus collected between I93o and I967 were compared in neuraminidase-inhibition and haemagglutination-inhibition serological tests. All the isolates were serologically related to a greater or lesser extent in tests with either antigen. The antigenicity of the two antigens varied independently in different isolates. There was a slight, but statistically significant,
Helicoverpa armigera Nuclear Polyhedrosis Virus (HearNPV) was isolated from the larva, obtained from four different geographic locations in India.\\u000a All the HearNPV isolates appeared clear, irregular, six sided in outline with rounded edges; phase bright under phase contrast\\u000a microscope; but varied in size, bioefficacy and restriction profiles. Anand isolate had largest mean diameter, highest frequency\\u000a of large polyhedral inclusion bodies
Charmi S. PatelJanardan; Janardan Jitendra Jani; Vipulkumar B. Parekh; Vijay B. Darji; Piyush R. Vaishnav
Hantaan (HTN) virus, the etiologic agent of clinically severe hemorrhagic fever with renal syndrome (HFRS), was first isolated in 1976 from lung tissue of a striped-field mouse (Apodemus agrarius) captured in Songnae-ri, Gyeonggi Province, Republic of Korea. Found primarily in mountainous areas, the Korean field mouse (A. peninsulae) is the second-most dominant field rodent species found throughout Korea. A new hantavirus, designated Soochong (SOO), was isolated in Vero E6 cells from four A. peninsulae captured in August 1997 at Mt. Gyebang in Hongcheon-gun, Mt. Gachil, Inje-gun, Gangwon Province, and in September 1998 at Mt. Deogyu, Muju-gun, Jeollabuk Province. The entire S, M, and L genomic segments of SOO virus, amplified by RT-PCR from lung tissues of seropositive A. peninsulae and from virus-infected Vero E6 cells, diverged from HTN virus (strain 76-118) by 15.6%, 22.8%, and 21.7% at the nucleotide level and 3.5%, 9.5%, and 4.6% at the amino acid level, respectively. Phylogenetic analyses of the nucleotide and deduced amino acid sequences, using the maximum parsimony and neighbor-joining methods, indicated that SOO virus was distinct from A. agrarius-borne HTN virus. SOO virus shared a common ancestry with Amur virus from Far East Russia, as well as with H5 and B78 hantaviruses, previously isolated from HFRS patients in China. Cross-focus-reduction neutralizating antibody tests showed that SOO virus, which is the first hantavirus isolated in cell culture from A. peninsulae, could be classified as a new hantavirus serotype. PMID:16372283
Baek, Luck Ju; Kariwa, Hiroaki; Lokugamage, Kumari; Yoshimatsu, Kumiko; Arikawa, Jiro; Takashima, Ikuo; Kang, Ju-Il; Moon, Sung Sil; Chung, Su Yong; Kim, Eun Ju; Kang, Hae Ji; Song, Ki-Joon; Klein, Terry A; Yanagihara, Richard; Song, Jin-Won
In 2003, H5N1 avian influenza virus infections were diagnosed in two Hong Kong residents who had visited the Fujian province in mainland China, affording us the opportunity to characterize one of the viral isolates, A/Hong Kong/213/03 (HK213; H5N1). In contrast to H5N1 virusesisolated from humans during the 1997 outbreak in Hong Kong, HK213 retained several features of aquatic bird viruses, including the lack of a deletion in the neuraminidase stalk and the absence of additional oligosaccharide chains at the globular head of the hemagglutinin molecule. It demonstrated weak pathogenicity in mice and ferrets but caused lethal infection in chickens. The original isolate failed to produce disease in ducks but became more pathogenic after five passages. Taken together, these findings portray the HK213 isolate as an aquatic avian influenza A virus without the molecular changes associated with the replication of H5N1 avian viruses in land-based poultry such as chickens. This case challenges the view that adaptation to land-based poultry is a prerequisite for the replication of aquatic avian influenza A viruses in humans. PMID:16014953
Orf virus is a parapoxvirus that causes recurring contagious ecthyma or orf disease in goat, sheep and other wild and domestic ruminants. Infected animals show signs of pustular lesions on the mouth and muzzle and develop scabs over the lesions. Although the infection is usually cleared within 1–2 months, delayed growth and associated secondary infections could still impact the herds. Orf virus can also infect humans, causing lesions similar to the animals in pathological histology. Prior infection of orf virus apparently offers little protective immunity against future infections. Several gene products of orf virus have been identified as responsible for immunomodulatory functions. In our recent study of orf virusisolates from an area along the Minjiang River in northern Fujian Province, we found a high heterogeneity among isolates from 10 farms within a 120-kilometer distance. Only two isolates from locations within 1 km to each other had same viral genes. There is no correlation between the geographical distance between the corresponding collection sites and the phylogenetic distance in ORFV011 or ORV059 genes for any two isolates. This finding suggests that there are diverse populations of orf virus present in the environment. This may in part contribute to the phenomenon of recurring outbreaks and heighten the need for better surveillance.
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
Dasheen mosaic virus (DsMV) is an important constraint to production of cocoyam (Xanthosoma spp.) in Nicaragua. Reverse transcription polymerase chain reaction was used to amplify the coat protein (CP) region from ten Nicaraguan DsMV isolates. These isolates showed high nucleotide identity to DsMV isolates from the USA, eastern Asia and Australasia. All Nicaraguan isolates except one shared a tandem repeat in the N-terminus of the CP. Phylogenetic analyses showed that the Nicaraguan isolates formed two distinct subgroups correlated with geographic origin. This can be explained by different origins of the cocoyam genotypes grown in these regions. PMID:19034605
Reyes, Guillermo; Ramsell, Jon N E; Nyman, Marie; Kvarnheden, Anders
A comparative study of the hemagglutinin (HA) receptor binding site (RBS) of a number of H13 influenza virusesisolated from Laridae family of birds (gulls) and other influenza viruses obtained from the Anatidae family (ducks) was conducted. The affinity of all viruses to alpha N-acetylneuraminic acid (Neu5Ac alpha), 3'sialyllactose (3'SL), and sialylglycopolymers bearing 3'-sialyl(N-acetyllactosamine) (3'SLN-PAA), [Neu5Ac alpha(2-3)Gal beta(1-4)][-Fuc alpha(1-3)]GlcNAc beta (SLe(x)-PAA), and [Neu5Ac alpha(2-3)Gal beta(1-3)][-Fuc alpha(1-4)]GlcNAc beta (SLe(a)-PAA), was determined. The last three polymer glycoconjugates were synthesized for determining the contribution of carbohydrate chains after the galactose link to the binding with the receptor. The difference in affinity between 3'SL and Neu5Ac alpha in all studied H13 viruses is small, which indicates a less significant role of the galactose moiety in the binding to the receptor. The results of virus binding with polymer sialylglycoconjugates indicates that the method of linking, the third monosaccharide moiety, and the presence of an extra fucose substitute in this moiety may influence the binding considerably. For virusesisolated from ducks, the suitable polymer is SLe(a)-PAA (i.e., a 1-3 linkage between galactose and glucosamine is optimal). This finding is in accord with the data that H13 virusesisolated from the gulls differ based on their ability to interact with polymer sialylglycoconjugates. The affinity to all three polymers is uniform, and the presence of GlcNAc-linked fucose does not prevent the binding. A comparative analysis of six sequenced HA H13 viruses and other subtype viruses showed presence of substantial differences in the composition of amino acids of this region in H13 viruses. PMID:14575135
Yamnikova, S S; Gambaryan, A S; Tuzikov, A B; Bovin, N V; Matrosovich, M N; Fedyakina, I T; Grinev, A A; Blinov, V M; Lvov, D K; Suarez, D L; Swayne, D E
Viruses of the family Bunyaviridae (the bunyaviruses) possess three distinct linear, single-stranded, negative sense or ambisense RNA segments (large, medium, and small). Dual infections of arthropod and perhaps vertebrate and plant hosts provide substantial opportunity for segment reassortment and an increasingly recognized number of the nearly 300 viruses in this family have been shown to be reassortants. Reassortment of RNA segments (genetic shift) complements genetic drift (accumulation of point mutations) as a powerful mechanism underlying bunyavirus evolution. Here we consider the possibility, if not likelihood, that most if not all bunyaviruses currently recognized may represent reassortants, some of which may be reassortants of existing viruses, and some of which may be reassortants of extinct viruses. If this hypothesis is correct, then the roots of the family and genus trees of bunyaviruses as currently described (or ignored) are incomplete or incorrect. PMID:24074583
Briese, Thomas; Calisher, Charles H; Higgs, Stephen
Monoclonal antibodies (mAbs) against a classical swine fever virus (CSFV; subgenogroup 1:1) isolate from Assam, India were produced and characterized. Four fusions of myeloma cells (SP2/0Ag) were made with spleenocytes of 8-10 weeks old BALB/C mice immunized with the viral antigen. Several hybridoma clones secreting antibodies to the virus were obtained after four fusions, but five hybridoma clones secreting antibody specific to the virus could be stabilized. All the mAbs belong to the IgG2a isotype. Except one, none of the four mAbs showed cross reaction with bovine viral diarrhoea virus and border disease virus (BDV). One mAb showed cross reaction with BDV. All the four mAbs specific to CSFV showed reactivity with the parental virus in immunoperoxidase test (IPT) and with a single protein band (molecular weight 55 kD approximately) of the virus in western blotting. In neutralization peroxidase linked assay (NPLA) all the mAbs reacted with 13 CSFV local isolates as well as with the cell culture adapted lapinized vaccine virus strain belonging to the subgenogroup 1:1. This is the first report on production and characterization of mAbs against CSFV in India. PMID:22822524
Three identical strains of an arbovirus were isolated from 475 Ornithodoros papillipes ticks collected in June, 1972, in burrows of the great gerbil (Rhombomys opimus Licht., 1882) in the environs of Beshkent, Karshinsk steppe, Uzbek S.S.R. The isolate was found to range among flaviviruses. Complement-fixation, agar diffusion precipitation and neutralization tests is tissue culture and mice indicated a one-way antigenic relationship between the isolate and West Nile virus. However, the pattern of differences between them made it possible to consider the isolated agent as a new virus, "Karshi" virus. The results of electron microscopic studies of this virus are presented. PMID:130853
Lvov, D K; Neronov, V M; Gromashevsky, V L; Skvortsova, T M; Berezina, L K; Sidorova, G A; Zhmaeva, Z M; Gofman, Y A; Klimenko, S M; Fomina, K B
Our aim was to establish the phylogenetic and genetic relationships among avian influenza viruses (AIV) recently isolated from poultry in Israel. During this study we analyzed complete nucleotide sequences of two envelope (hemagglutinin and neuraminidase) and six internal genes (polymerase B1, polymerase B2, polymerase A, nucleoprotein, nonstructural, and matrix) of 29 selected H9N2 and six internal genes of five H5N1 virusesisolated in Israel during 2000-2006. Comparative genetic and phylogenetic analyses of these sequences revealed that the local H5N1 viruses are closely related to H5N1 virusesisolated in European, Asian, and Middle Eastern countries in 2005-2006. The H9N2 Israeli isolates, together with virusesisolated in Jordan and Saudi Arabia formed a single group. Our data support the claim that during recent years a new endemic focus of H9N2 has been formed in the Middle East. The introduction of H5N1 and co-circulation of these two subtypes of AIV in this region may augment the risk of potentially pandemic strains emergence. PMID:18712589
Summary. Sequence was determined for the coat protein (CP) gene and 3? non-translated region (3?NTR) of two vanilla mosaic virus (VanMV)\\u000a isolates from Vanilla tahitensis, respectively from the Cook Islands (VanMV-CI) and French Polynesia (VanMV-FP). Both viruses displayed distinctive features\\u000a in the N-terminal region of their CPs; for VanMV-CI, a 16-amino-acid deletion including the aphid transmission-related DAG\\u000a motif, and for VanMV-FP,
K. Farreyrol; M. N. Pearson; M. Grisoni; D. Cohen; D. Beck
An H5N1 influenza A virus was isolated from duck meat processed for human consumption, imported to Japan from Shandong Province, China in 2003. This virus was antigenically different from other H5 viruses, including the Hong Kong H5N1 virusesisolated from humans in 1997 and 2003. Sequence analysis revealed that six genes (PB1, PA, HA, NA, M, and NS) of this virus showed >97% nucleotide identity with their counterparts from recent H5N1 viruses, but that the remaining two genes (PB2 and NP) were derived from other unknown viruses. This duck meat isolate was highly pathogenic to chickens upon intravenous or intranasal inoculation, replicated well in the lungs of mice and spread to the brain, but was not as pathogenic in mice as H5N1 human isolates (with a dose lethal to 50% of mice (MLD50)=5x10(6) 50% egg infectious doses [EID50]). However, virusesisolated from the brain of mice previously infected with the virus were substantially more pathogenic (MLD50=approximately 10(2) EID50) and possessed some amino acid substitutions relative to the original virus. These results show that poultry products contaminated with influenza viruses of high pathogenic potential to mammals are a threat to public health even in countries where the virus is not enzootic and represent a possible source of influenza outbreaks in poultry. PMID:15964604
Pigs are proposed to be "mixing vessel" hosts that can produce genetically novel reassortant viruses with pandemic potential. The appearance of any novel influenza viruses among pigs should pose concerns for human health. Here, we report the complete genome sequence of a novel H4N1 influenza virus [A/Swine/HuBei/06/2009(H4N1)] isolated from a pig in Central China in 2009. The genomic sequence analysis indicates that this virus is a wholly avian-original influenza virus. Each gene may come from different avian influenza viruses outside mainland China, suggesting the role of migratory birds in the dispersal of influenza virus. PMID:23166273
Viruses have been hypothesized to control blooms of Aureococcus anophagefferens gen. et sp. nov. (Chrysophyceae), a marine phytoplankton that since 1985 has caused devastating summer blooms called "brown tide." By means of ultrafiltration methods, viruses specific to this alga were isolated from both the Great South Bay and Peconic Bay systems of Long Island, New York, during the summer bloom period of 1992. Cell lysis of healthy algal cultures was demonstrated, as well as continuing reinfection with serial transfers of cultures. Electron microscope surveys yielded images of phage-like virus particles with tails that could attach to A. anophagefferens cells within minutes of exposure. The isolation and cultivation of this virus highlights the need for further study of viral infection of eukaryotic algae and the potential for a better understanding of algal bloom control by viral infection. PMID:17730401
Partial genome segments of a begomovirus were previously amplified from Wissadula amplissima exhibiting yellow-mosaic and leaf-curl symptoms in the parish of St. Thomas, Jamaica and this isolate assigned to a tentative begomovirus species, Wissadula golden mosaic St. Thomas virus. To clone the complete genome of this isolate of Wissadula golden mosaic St. Thomas virus, abutting primers were designed to PCR amplify its full-length DNA-A and DNA-B components. Sequence analysis of the complete begomovirus genome obtained, confirmed that it belongs to a distinct begomovirus species and this isolate was named Wissadula golden mosaic St. Thomas virus-[Jamaica:Albion:2005] (WGMSTV-[JM:Alb:05]). The genome of WGMSTV-[JM:Alb:05] is organized similar to that of other bipartite Western Hemisphere begomoviruses. Phylogenetic analyses placed the genome components of WGMSTV-[JM:Alb:05] in the Abutilon mosaic virus clade and showed that the DNA-A component is most closely related to four begomovirus species from Cuba, Tobacco leaf curl Cuba virus, Tobacco leaf rugose virus, Tobacco mottle leaf curl virus, and Tomato yellow distortion leaf virus. The putative Rep-binding-site motif in the common region of WGMSTV-[JM:Alb:05] was observed to be identical to that of Chino del tomate virus-Tomato [Mexico:Sinaloa:1983], Sida yellow mosaic Yucatan virus-[Mexico:Yucatan:2005], and Tomato leaf curl Sinaloa virus-[Nicaragua:Santa Lucia], suggesting that WGMSTV-[JM:Alb:05] is capable of forming viable pseudo-recombinants with these begomoviruses, but not with other members of the Abutilon mosaic virus clade. Biolistic inoculation of test plant species with partial dimers of the WGMSTV-[JM:Alb:05] DNA-A and DNA-B components showed that the virus was infectious to Nicotiana benthamiana and W. amplissima and the cultivated species Phaseolus vulgaris (kidney bean) and Lycopersicon esculentum (tomato). Infected W. amplissima plants developed symptoms similar to symptoms observed under field conditions, confirming that this virus is a causal agent of Wissadula yellow mosaic disease in W. amplissima. PMID:19768650
Collins, A M; Mujaddad-ur-Rehman, Malik; Brown, J K; Reddy, C; Wang, A; Fondong, V; Roye, M E
Genetic analysis was performed on 13 hepatitis D virus (HDV) isolates from Ethiopia, Somalia, Jordan, Kuwait, Bulgaria, Moldavia, and Sweden. The complete nucleotide sequence and genomic organization are described for the first time for two African HDV isolates. Phylogenetic analysis showed all the African isolates to be intrarelated and to form a novel group within HDV genotype I; the suggested designation for this group is IC. The genetic distance to previously described type I isolates was about 0.15. The HDV genotype I isolates (total of 22 examined) phylogenetically formed three clusters, each of them corresponding to certain geographic regions; the "western" group consisted of six HDV isolates from western Europe and the United States plus one from Kuwait; the "eastern" group consisted of two isolates from Moldavia and one each from Bulgaria, Nauru, mainland China, and Taiwan; and the "African-Middle East" group consisted of six HDV isolates from Ethiopia and one from Somalia, Jordan, and Lebanon.
We characterized low pathogenic avian influenza (LPAI) H5N2 and H9N2 virusesisolated in South Korea from 2008 to 2009. Genetic analysis of the H5N2 virusesisolated from wild birds and domestic ducks demonstrated that they were related to the recently isolated southern Chinese LPAI H5 viruses and various influenza viruses circulating in Eurasia. Three H9N2 viruses obtained at live bird markets and duck farms were reassortant viruses generated from the H5N2 viruses of domestic ducks and the H9N2 virus endemic in Korean chickens. The H5N2 viruses did not replicate well in experimentally infected chickens and mice, but novel H9N2 viruses, without pre-adaptation, were recovered at high titres in chickens. Our results show that reassortment between H5N2 and H9N2 viruses must have occurred in domestic ducks and may have contributed to the diversity expansion of the gene pool, which has potential to alter the pathogenicity and host range of the influenza virus. PMID:20392898
Kim, Hye-Ryoung; Park, Choi-Ku; Oem, Jae-Ku; Bae, You-Chan; Choi, Jun-Gu; Lee, O-Soo; Lee, Youn-Jeong
We describe here serological and structural properties of a virus, Mason-Pfizer Monkey Virus (M-PMV), isolated from a simian mammary tumor. This virus is morphologically similar to the known oncogenic RNA viruses (oncornaviruses). It has a 60-70S RNA, and its replication is inhibited by actinomycin. Antisera prepared against the virusisolated by density-gradient centrifugation identify at least two viral structural antigens. Immunodiffusion studies show that this virus is serologically unrelated to three types of simian foamy viruses, visna virus, and the known oncornaviruses. Immunofluorescence reveals that the structural proteins of the virus are synthesized cytoplasmically. Although M-PMV productively infects human cells in vitro, serological analysis does not show the presence of M-PMV antigens in human neoplasia. Images
Nowinski, Robert C.; Edynak, Eugene; Sarkar, Nurul H.
The 2004 outbreaks of highly pathogenic avian influenza H5N1 disease in China led to a great poultry loss and society attention. A survey of avian influenza viruses was conducted on tree sparrows (Passer montanus) collected in China in 2004. Four viruses were isolated from free-living tree sparrows. The results of the whole-genome analysis indicated that an H5N1 virus with a new genotype is circulating among tree sparrows. The hemagglutinin and neuraminidase genes of the new genotype were derived from Gs/Gd/96-like viruses and the nuclear protein gene descended from the 2001 genotype A H5N1 viruses, while the other inner genes originated from an unknown influenza virus. In experimental infection, all four viruses were highly pathogenic to chickens but not pathogenic to ducks or mice. The four tree sparrow viruses were different from the 2003 tree sparrow strain (genotype Z) in Hong Kong. The results suggested that H5N1 viruses might be distributed widely in tree sparrows. PMID:16306617
Kou, Z; Lei, F M; Yu, J; Fan, Z J; Yin, Z H; Jia, C X; Xiong, K J; Sun, Y H; Zhang, X W; Wu, X M; Gao, X B; Li, T X
Studies indicate that West African and Congo basin isolates of monkeypox virus (MPXV) are genetically distinct. Here, we show Congo basin MPXV-ZAI-V79 is more virulent for cynomolgus monkeys as compared to presumed West African MPXV-COP-58. This finding may explain the lack of case-fatalities in the U.S. 2003 monkeypox outbreak, which was caused by a West African virus. Virulence differences between
Nanhai Chen; Guiyun Li; M. Kathryn Liszewski; John P. Atkinson; Peter B. Jahrling; Zehua Feng; Jill Schriewer; Charles Buck; Chunlin Wang; Elliot J. Lefkowitz; Joseph J. Esposito; Tiara Harms; Inger K. Damon; Rachel L. Roper; Chris Upton; R. Mark L. Buller
The genome of a Spanish isolate of Parietaria mottle virus (PMoV) obtained from tomato (strain PMoV-T) was completely sequenced. Protein motifs conserved for RNA viruses were identified:\\u000a the p1 protein contained a metyltransferase domain in its N-terminal half and a triphosphatase\\/helicase domain in its C-terminal\\u000a half, the p2 protein contained a RNA polymerase domain; the 3a protein contained a RNA-binding
Luis Galipienso; Luis Rubio; Carmelo López; Salvador Soler; José Aramburu
Abstract. La Crosse virus (LACV) is found,primarily in the Midwestern,and,Appalachian regions of the United States where,it is a leading cause of mosquito-borne,encephalitis in children. To determine,whether,the distribution of this virus extends further east into New England, we analyzed a bunyavirus that was isolated from a pool of eastern tree-hole mosquitoes, Ochlerotatus triseriatus ( Aedes triseriatus), collected from Fairfield, Connecticut (CT)
Summary. 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\\u000a isolate) antiserum. Antiserum raised to the Chinese virus did not react with eighteen other potyviruses. The complete nucleotide\\u000a sequence (9816?nt) had the typical
J. Chen; Y.-H. Shi; M. J. Adams; H.-Y. Zheng; B.-X. Qin; J.-P. Chen
The characterization of an orf virus (OV) isolated from skin lesions of a goat kid with severe, persistent, proliferative dermatitis, and designated orf virus-San Angelo 2000 (OV-SA00) strain, is described. The identity of OV-SA00 was confirmed by a combination of methods, including electron microscopy, amplification of specific fragments of viral DNA by polymerase chain reaction, restriction enzyme analysis of viral
J. Guo; Z. Zhang; J. F. Edwards; R. W. Ermel; C. Taylor; A. de la Concha-Bermejillo
Eleven infectious bursal disease virus (IBDV) strains isolated recently from China were compared with the early classical virulent strain CJ801, the chicken embryo fibroblast-adapted (CEF) variant strain GZ902, and the attenuated vaccine strains BJ836, BK912, and LM to discern the evolutionary characteristics of IBDV in China at both antigenic and genetic levels. Virus neutralization (VN) assay showed that all ten
It has been estimated that, to date, about 48% of all HIV-infected people in the world carry HIV-1 subtype C virus. Therefore, it is of great importance to gain better knowledge about the genetic and biological characteristics of this virus subtype. In the present study, the biological properties of HIV-1 isolates obtained from nine Ethiopian patients with AIDS were studied. DNA sequencing of the V3 loop of gp120 classified the isolates as subtype C. In primary isolation cultures, virus infection was accompanied by syncytium formation and cell lysis. Interestingly, when examining the growth in primary monocyte-macrophage cultures, initial low-level virus replication was followed by a nonproductive state, from which virus could be rescued by cocultivation with Jurkat(tat) cells. Furthermore, none of the isolates replicated in T cell lines (CEM, MT-2, HuT-78, and H9) or in the promonocytic cell line U937 clone 2. All isolates could use CCR5 as coreceptor, whereas no isolates could use CCR2b, CCR3, CCR5, CXCR4, Bonzo/STRL33, or BOB/GPR15. The genotype of the V3 region correlated with the MT-2 negative/non-syncytium-inducing (NSI) phenotype. Comparative studies revealed that the scarcity of CXCR4 usage as well as other phenotypic characteristics of subtype C isolates distinguish this subtype. On the basis of these data, we suggest that in addition, factors other than viral phenotype may govern the pathogenic potential of subtype C isolates. PMID:10331443
Björndal, A; Sönnerborg, A; Tscherning, C; Albert, J; Fenyö, E M
Following howling monkey (Alouatta caraya) deaths and yellow fever (YF) antigen detection by immunohistochemistry in the liver sample of a dead monkey in April and May 2001 in the municipalities of Garruchos and Santo Antônio das Missőes, Rio Grande do Sul State, Brazil, epidemiological field investigations were initiated. Two strains of YF virus were isolated in suckling mice from 23 Haemagogus (Conopostegus) leucocelaenus Dyar & Shannon mosquitoes collected from the study sites. The YF virus was isolated from this species in the 1930s in Brazil and in the 1940s in Colombia. No human cases were reported during the current epizootic outbreak. The YF virusisolation and the absence of Hg. (Haemagogus) janthinomys Dyar from the area suggest that Hg. leucocelaenus may be a secondary YF vector and play an important role in the epidemiology of this disease in the Southern Cone. PMID:12892055
Vasconcelos, Pedro F; Sperb, Alethéa F; Monteiro, Hamilton A; Torres, Maria A; Sousa, Maria R; Vasconcelos, Helena B; Mardini, Lúcia B; Rodrigues, Sueli G
A total of 1,397 rodents of 7 different species were collected in the major vegetative zones of Senegal. Organ pools from rodents were inoculated into suckling-mice. Thirty-five viral strains, representing 5 viruses, were isolated. The 5 viral types recovered in the present survey comprise: Bandia (23 strains), Saboya (7), Salanga (3), Gabek Forest (1) and a new poxvirus (AnD 42332) for which the name of Fadiga is proposed. This virus was isolated from a Mastomys sp collected in eastern Senegal. Antigenic relations were established by complement fixation test with Salanga virus; its distinctness was determined by neutralisation test. Our data are discussed with compiled informations on the current status of rodent viral isolations in West and Central Africa. PMID:3094972
A virus, 78-238, isolated from the cerebrospinal fluid of a dog with neurological dysfunction, was characterized as a paramyxovirus. This conclusion was supported by viral cytopathic effects and morphological appearance of virions and nucleocapsids in infected cells. Nucleocapsids were found in the cytoplasm of all infected cells and in the nuclei of 0.001% of these cells. Growth curves revealed that a high percentage (?76%) of infectious progeny virus was cell released. Persistent infection of Vero cells with 78-238 showed a consistently high percentage of fluorescence-positive cells and a low proportion of hemadsorption-positive cells. Serological studies indicate that the virus was closely related to Simian virus 5 and reference canine parainfluenza virus. Images
Baumgartner, Wolfgang K.; Metzler, Alfred E.; Krakowka, Steven; Koestner, Adalbert
South Korean isolates of oseltamivir-resistant influenza viruses from 2005-2010 were investigated with a total 491 influenza viruses identified from 1702 specimens. Neuraminidase genes from 342 influenza viruses (71 A/H1N1, 74 pandemic A/H1N1 2009, 117 A/H3N2, and 80 B) were analyzed by RT-PCR with molecular markers for oseltamivir resistance. The H274Y mutation in the NA protein was identified in 100 % (n=40) of A/H1N1 viruses circulating in 2008-2009. Influenza A/H1N1 viruses harboring the H274Y substitution exhibited, on average, a 626-fold reduction in oseltamivir susceptibility and clustered with the A/Norway/1736/2007 strain. Close and timely monitoring for resistance to clinically available influenza antivirals should be consistently performed. PMID:23690054
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.
Leanyer virus (LEAV), currently classified as a member of the genus Orthobunyavirus, in the family Bunyaviridae, was originally isolated from a pool of Anopheles meraukensis mosquitoes, collected at Leanyer, Northern Territory, Australia in 1974. When it failed to react in serological tests with antisera from other known viruses, full-length genomic sequencing was pursued to determine the relationship of LEAV to other orthobunyavirus species. Genetic and serological characterization confirmed its antigenic distance from other orthobunyaviruses, including to its closest genetic neighbours, the Simbu group viruses, suggesting that it may represent a new antigenic complex.
Savji, Nazir; Travassos da Rosa, Amelia; Hutchison, Stephen; Celone, Christopher; Hui, Jeffrey; Briese, Thomas; Calisher, Charles H.; Lipkin, W. Ian
A novel human B-lymphotropic virus (HBLV) was isolated from the peripheral blood leukocytes of six individuals: two HTLV-III seropositive patients from the United States (one with AIDS-related lymphoma and one with dermatopathic lymphadenopathy), three HTLV-III seronegative patients from the United States (one with angioimmunoblastic lymphadenopathy, one with cutaneous T-cell lymphoma, and one with immunoblastic lymphoma), and one HTLV-III seronegative patient with acute lymphocytic leukemia from Jamaica. All six isolates were closely related by antigenic analysis, and sera from all six virus-positive patients reacted immunologically with each virusisolate. In contrast, only four sera from 220 randomly selected healthy donors and none from 12 AIDS patients without associated lymphoma were seropositive. The virus selectively infected freshly isolated human B cells and converted them into large, refractile mono- or binucleated cells with nuclear and cytoplasmic inclusion bodies. HBLV is morphologically similar to viruses of the herpesvirus family but is readily distinguishable from the known human and nonhuman primate herpesviruses by host range, in vitro biological effects, and antigenic features. PMID:2876520
Salahuddin, S Z; Ablashi, D V; Markham, P D; Josephs, S F; Sturzenegger, S; Kaplan, M; Halligan, G; Biberfeld, P; Wong-Staal, F; Kramarsky, B
Characterization of field isolates of viruses associated with pox-like outbreaks involving both cows (cattle) and buffaloes\\u000a was carried out. PCR and electron microcopy of representative virusisolates from these animals, initially identified them\\u000a as orthopoxviruses (OPXVs). Sequence and phylogenetic analyses of A-type inclusion and haemagglutinin (HA) genes of these\\u000a isolates revealed a closer relationship with other OPXVs. Sequencing of the
S. Yadav; M. Hosamani; V. Balamurugan; V. Bhanuprakash; Raj Kumar Singh
The results of the isolation and identification of the causative agent of a haemorrhagic fever outbreak in the Stavropol Territory are presented. The virusisolated from blood of haemorrhagic fever patients by virological methods was identified in serological and molecular tests as Crimean haemorrhagic fever virus. This epidemiological analysis testify to increased activity of the natural focus of Crimean-Congo haemorrhagic fever in this area due to a number of natural and other factors leading to intensification of its epidemic realization. PMID:11881502
Onishchenko, G G; Markov, V I; Merkulov, V A; Vasil'ev, N T; Berezhno?, A M; Androshchuk, I A; Maksimov, V A
A street rabies virus (RV) isolate, GXHXN, was obtained from brain tissue of rabid cattle in the Guangxi Zhuang Autonomous Region of China in 2009. GXHXN is the first isolate from cattle in China with its entire genome sequenced and is closely related to BJ2011E from horse in Beijing, WH11 from donkey in the Hubei Province, and isolates from dogs in the Guangxi and Fujian Provinces, with homologies of 97.6% to 99.6%. It is more distantly related to isolates from domestic cat, pig, Chinese ferret badger, and vaccine strains, with homologies of 83.1% to 88.0%.
Aptamers, functional nucleic acids, capable of binding a variety of molecular targets with high affinity and specificity, have emerged as promising therapeutic agents. In this study, the cell surface-systematic evolution of ligands by exponential enrichment (Cell-SELEX) strategy was used to generate DNA aptamers which targeted to the intact rabies virus-infected live cells. Through 35 iterative rounds of selection, five high-affinity single-stranded DNA (ssDNA) aptamers were generated by cell-SELEX. Virus titer assay and real-time quantitative reverse transcription PCR (qRT-PCR) assay revealed that all five aptamers could inhibit replication of rabies virus (RABV) in cultured baby hamster kidney (BHK)-21 cells; and T14 and F34 aptamers were most effective. The qRT-PCR also showed a dose-dependent inhibitory effect in BHK-21 cells. Collectively, these data show the feasibility of generating functionally effective aptamers against rabies virus-infected cells by the Cell-SELEX iterative procedure. These aptamers may prove clinically useful as therapeutic molecules with specific antiviral potential against RABV infections. PMID:22771543
Beak and feather disease virus (BFDV), the causative agent of psittacine beak and feather disease (PBFD) infects psittaciformes worldwide. We provide an annotated sequence record of three full-length unique genomes of BFDV isolates from budgerigars (Melopsittacus undulatus) from a breeding farm in South Africa. The isolates share >99% nucleotide sequence identity with each other and approximately 96% nucleotide sequence identity to two recent isolates (Melopsittacus undulatus) from Thailand but only between 91.6 and 86.6% identity with all other full-length BFDV sequences. Maximum-likelihood analysis and recombination analysis suggest that the South African budgerigar BFDV isolates are unique to budgerigars, are non-recombinant in origin, and represent a new genotype of BFDV. PMID:20127375
Varsani, Arvind; de Villiers, Gillian K; Regnard, Guy L; Bragg, Robert R; Kondiah, Kulsum; Hitzeroth, Inga I; Rybicki, Edward P
African swine fever is widespread in Africa but has occasionally been introduced into other continents. In June 2007, African swine fever was isolated in the Caucasus Region of the Republic of Georgia and subsequently in neighboring countries (Armenia, Azerbaijan, and 9 states of the Russian Federation). Previous data for sequencing of 3 genes indicated that the Georgia 2007/1 isolate is closely related to isolates of genotype II, which has been identified in Mozambique, Madagascar, and Zambia. We report the complete genomic coding sequence of the Georgia 2007/1 isolate and comparison with other isolates. A genome sequence of 189,344 bp encoding 166 open reading frames (ORFs) was obtained. Phylogeny based on concatenated sequences of 125 conserved ORFs showed that this isolate clustered most closely with the Mkuzi 1979 isolate. Some ORFs clustered differently, suggesting that recombination may have occurred. Results provide a baseline for monitoring genomic changes in this virus. PMID:21470447
Chapman, David A G; Darby, Alistair C; Da Silva, Melissa; Upton, Chris; Radford, Alan D; Dixon, Linda K
Restriction Fragment Length Polymorphism (RFLP) technique was employed to investigate the relationship of Pakistani isolates of banana bunchy top virus (BBTV) with Asia and South Pacific strains. The PCR amplified product of BBTV DNA component I was digested with nine different restriction endonucleases (Bam HI, EcoRI, EcoRV, HaeIII, HincII, HindIII, PstI, RsaI and SmaI) and analyzed by agarose gel electrophoresis.
TAHIRA YASMIN; S. M. SAQLAIN NAQVI; HUSSAIN SHAH; SAIF KHALID
Highly pathogenic avian influenza (HPAI) viruses of the H5N1 subtype have caused several rounds of outbreaks in Thailand. In this study, we used 3 HPAI virusesisolated in Thailand in January 2004 from chicken, quail, and duck for genetic and pathogenetic studies. Sequence analysis of the entire genomes of these isolates revealed that they were genetically similar to each other.
We analyzed 50 rabies virus samples isolated in Brazil from 12 dogs, 11 cats, 5 vampire bats, 15 cattle, 2 horses, 1 pig, 1 sheep, and 3 humans to investigate the molecular epidemiology of rabies viruses. We sequenced 203 nucleotides on the nucleoprotein gene by direct sequencing of the PCR-amplified products. All the isolates belonged to the genotype 1 and
Mikako Ito; Yohko T. Arai; Takuya Itou; Takeo Sakai; Fumio H. Ito; Tomohiko Takasaki; Ichiro Kurane
This paper describes the isolation of porcine epidemic diarrhea (PED) virus in Vero and porcine cell cultures, and the influence of age on disease in experimental infection. PED virus was isolated from the small intestine of piglets inoculated with PED samples and cultured in Vero, porcine bladder and kidney cells propagated in collagen-coated tissue culture plates in maintenance medium (MM)
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
We tested the amantadine-resistance among avian influenza A (H5N1) virusesisolated from chicken in Hebei Province of Northern China from 2001 to 2005, and investigated the amantadine use in this area. Plague reduction assay in MDCK cells showed that 83.3% isolates (5/6) were amantadine-resistant strains. The M2 sequence analysis revealed that four of five resistant isolates contained the point mutations (Ser to Asn) at position 31 that could confer resistance to amantadine. These results indicated that the incidence of amantadine-resistant virusesisolated in Northern China was particularly high. In the investigation of amantadine use, we found that amantadine was used extensively in poultry farms in this area, which maybe was one of reasons of the high amantadine-resistance incidence. PMID:17897729
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.
Complete nucleotide sequences of the DNA-A-like molecules of three East African cassava mosaic virus (EACMV) isolates from Kenya (-K, 2801 nt) and Malawi (-MH and -MK, both 2804 nt) were determined. These sequences were compared with that published for a Tanzanian isolate (-T, 2801 nt) and the partial sequence of a third Malawian isolate. Intergenic region sequences of all isolates,
Xueping Zhou; David J. Robinson; Bryan D. Harrison
On December 7, 2010, H5N1 highly pathogenic avian influenza virus was isolated from a healthy mallard captured at the Mankyung River in South Korea. Phylogenetic analysis showed that this virus was classified into clade 2.3.2 and closely related to H5N1 virusesisolated from wild birds in Mongolia, Russia and China in 2009 and 2010. PMID:21466927
Kim, Hye-Ryoung; Kim, Bang-Sil; Bae, You-Chan; Moon, Oun-Kyoung; Oem, Jae-Ku; Kang, Hyun-Mi; Choi, Jun-Gu; Lee, O-Soo; Lee, Youn-Jeong
Classical swine fever virus (CSFV) causes a highly contagious disease that leads to significant economic losses in the pig\\u000a industry worldwide. However, there is a paucity of knowledge on the accurate genotyping of CSFV isolates in south China. This\\u000a study genotyped the E2 gene of 14 CSFV strains isolated during 2008–2010 from domestic pigs in different districts of south\\u000a China.
Recently, three distinct genotypes of clinical herpes simplex virus type 1 (HSV-1) isolates were identified based on DNA sequence information and phylogenetic analysis of clinical isolates and laboratory strains. We utilized single-nucleotide polymorphism within the genes coding for glycoproteins G and I for rapid genotype classification by PCR and restriction enzyme cleavage. The method is suitable for high-scale genotyping of
Peter Norberg; Tomas Bergstrom; Jan-Ĺke Liljeqvist
Background Bovine viral diarrhea virus (BVDV) infections continue to cause significantly losses in the deer population. Better isolation\\u000a and identification of BVDV from sika deer may contribute significantly to the development of prophylactic therapeutic, and\\u000a diagnostic reagents as well as help in prevention and control of BVDV. However, isolation and identification of BVDV from\\u000a sika deer is seldom reported in literature.
Yugang Gao; Shijie Wang; Rui Du; Quankai Wang; Changjiang Sun; Nan Wang; Pengju Zhang; Lianxue Zhang
Summary. ?Six recently isolated field strains of infectious bursal disease virus (IBDV) were compared to vaccine strains at the antigenic\\u000a and genetic level to ascertain the level of heterogeneity among Australian IBDV strains. Five strains, 01\\/94, 02\\/95, 03\\/95,\\u000a 04\\/95 and 08\\/95, isolated at four locations in the state of Victoria, were antigenic variants. They failed to react with monoclonal\\u000a antibodies directed
Summary. ?Seven neutralizing monoclonal antibodies (Mabs) to infectious bursal disease virus (IBDV) were used in an antigen-capture\\u000a ELISA for the antigenic characterization of 58 IBDV isolates obtained in France since 1989. Fifty-six isolates exhibited an\\u000a antigenic profile which was different from reference strain Faragher 52\\/70, and similar to French very virulent IBDV strain\\u000a 89?163 (no binding of two Mabs). Two strains
N. Eterradossi; G. Rivallan; D. Toquin; M. Guittet
A widespread porcine epidemic diarrhea virus (PEDV) occurred in southern China during 2010 to 2012. A virulent field PEDV strain, GD-B, was isolated from a sucking piglet suffering from severe diarrhea in Guangdong, China. We sequenced and analyzed the complete genome of strain GD-B, which will promote a better understanding of the molecular epidemiology and genetic diversity of PEDV field isolates in southern China. PMID:22879620
Summary Three virus strains isolated fromIxodes putus ticks were shown to be related to, though not identical, with Uukuniemi virus by complement-fixation tests. No antigenic relations were detected in the cross-neutralization test performed with the virusisolated and Uukuniemi virus. Two virus strains were isolated in 1969 on Tyuleniy island, Zaliv Terpeniya (Patience Bay) of the Sea of Okhotsk (Sakhalin
D. K. Lvov; A. A. TIMOPI-IEEVA; V. L. GROMASttEVSKI; G. V. GOSTINSIICIIIKOVA; O. V. Veselovskaya; V. I. Chervonski; K. B. Fomina; A. I. Gromov; A. G. Pogrebenko; V. Yu. Zhezmer
A purified antigen, HABA protein, has been derived from influenza virus concentrates by extraction with denaturing solvents. The protein lacks hemagglutinating activity but binds completely strain-specific, hemagglutination-inhibiting antibodies and induces neutralizing antibodies in experimental animals. Physicochemical characterization of HABA protein identifies it as a single homogeneous glycoprotein with a molecular weight of 78,000. On dissociation with guanidine or sodium dodecyl sulfate, in the presence of reducing agents, only one size of polypeptide with a molecular weight of the order of 40,000 is characteristic of the preparations. The data indicate that HABA protein is a dimer of HA1 polypeptide of the influenza virus hemagglutinin substructure, and that only trace amounts of other polypeptides are present.
Ten 3-month-old Tibetan mastiffs became ill 2 days after they were bought from a Tibetan mastiff exhibition, and 4 of them died 2 weeks later. A canine influenza virus (ZJ0110) was isolated from the lung of a deceased Tibetan mastiff and was characterized in detail. Sequence analysis indicated that the 8 genes of the canine isolate were most similar to those of avian-origin canine influenza viruses (H3N2) isolated in South Korea in 2007, with which they shared >98% sequence identity. ZJ0110 could experimentally infect 6-month-old beagles by intranasal inoculation and by airborne transmission, causing severe respiratory syndrome. Moreover, ZJ0110 could replicate in the upper respiratory tracts of mice and guinea pigs, and the virus titer was comparable to that in the upper respiratory tracts of dogs. Although the virus was genetically of avian origin, ZJ0110 could not experimentally infect chicken or ducks by intranasal inoculation. These results suggest that dogs might be an intermediary host in which avian influenza viruses adapt to replicate in mammals. PMID:23107656
Sequential isolates from eight cynomolgus monkeys experimentally infected with simian immunodeficiency virus (SIVsm, of sooty mangabey origin) were tested for coreceptor use in the human osteosarcoma indicator cell line, GHOST(3), expressing CD4 and one or another of the chemokine receptors CCR3, CCR5, CXCR4, BOB, or the orphan receptor Bonzo. The indicator cell line carries the human immunodeficiency virus type 2 long terminal repeat-driven green fluorescence protein gene that becomes activated upon infection with HIV or SIV and fluorescence can be quantitated by flow cytometric analysis. The methodological details are described in the accompanying paper (Vödrös et al., 2001, Virology 290, in press). All SIVsm inoculum viruses and reisolates used CCR5 with a high level of efficiency. CCR5 use was stable over time. BOB and Bonzo use was less efficient than CCR5 use and, in particular, late isolates obtained at the time of immunodeficiency varied greatly in their coreceptor use and often could not establish a productive infection in BOB- or Bonzo-expressing cells. Unexpectedly, early reisolates obtained 12 days postinfection could infect the entire GHOST(3) panel including the parental cells. In one case this was due to use of CXCR4, either transfected or endogenously expressed on the GHOST(3) cells. Our results demonstrate the complex coreceptor use of SIVsm isolates. Moreover, they focus attention on the initial phase of virus replication when the availability of target cells may govern the replication pattern of the virus. PMID:11878872
Vödrös, D; Thorstensson, R; Biberfeld, G; Schols, D; De Clercq, E; Fenyö, E M
Equine-virulent, epidemic\\/epizootic strains of Venezuelan equine encephalitis (VEE) virus (VEEV) arise via mutation of progenitor enzootic strains that replicate poorly in equines. Sequencing studies have implicated positively charged amino acids on the surface of the E2 envelope glycoprotein in the acquisition of equine virulence and viremia potential, suggesting that changes in binding to cell surface glycosaminoglycans (GAGs) may mediate VEE
Eryu Wang; Aaron C. Brault; Ann M. Powers; Wenli Kang; Scott C. Weaver
Summary. LMV-Common and LMV-Most are two seed-borne types of Lettuce mosaic virus (LMV), genus Potyvirus. LMV-Most, but not LMV-Common, overcomes the resistance afforded to lettuce by two recessive genes, mo1 1 and mo1 2. An RT-PCR-based assay thought to be specific for LMV-Most also amplified LMV-Tn2, previously typified as LMV-Common. The sequence of selected regions along the genome indicated that
R. Krause-Sakate; H. Fakhfakh; M. Peypelut; M. A. Pavan; F. M. Zerbini; M. Marrakchi; T. Candresse; O. Le Gall
Summary Three viruses were isolated from ticks parasitizing nesting seabirds on Cape Sizun, Brittany, France, during the spring of 1979: Soldado like virus (Bunyaviridae, Nairovirus genus) fromOrnithodoros (Alectorobius) maritimus (Argasidae), Zaliv Terpeniya (ZT) virus (Bunyaviridae, Uukuvirus genus) fromIxodes (Ceratixodes) uriae (Ixodidae) parasitizing chicks of the kittiwake(Rissa tridactyla), and a virus of the Sakhalin group fromI. (C.) uriae. Certain virological and
C. CtIASTEL; J. Y. Monnat; G. Le Lay; C. Guiguen; M. C. Quillien; J. C. Beaucournu
Since the 1997 H5N1 influenza virus outbreak in humans and poultry in Hong Kong, the emergence of closely related viruses in poultry has raised concerns that additional zoonotic transmissions of influenza viruses from poultry to humans may occur. In May 2001, an avian H5N1 influenza A virus was isolated from duck meat that had been imported to South Korea from
Terrence M. Tumpey; David L. Suarez; Laura E. L. Perkins; Dennis A. Senne; Jae-gil Lee; Youn-Jeong Lee; In-Pil Mo; Haan-Woo Sung; David E. Swayne
A noncytopathic type 1a bovine viral diarrhea virus (BVDV) was isolated from a free-ranging yearling female mule deer (Odocoileus hemionus) from northwestern Wyoming (USA). The mule deer was emaciated, weak, and salivating, and Arcanobacterium pyogenes was cultured from lung abscesses. Bovine viral diarrhea virus was isolated from lung, however, BVDV antigen was not detected by immunohistochemistry. The BVDV genotype was determined by reverse transcriptase polymerase chain reaction and the RNA sequences from the 5'UTR and E2 genes compared with sequences of a type 1a BVDV isolated from cattle from the same area as the deer. The sequences from the deer BVDV were distinct from those of the bovine type 1a BVDV, but similar to other bovine type 1a BVDVs. Seventy-four (60%) of 124 sera collected from mule deer in this area had serum neutralizing antibody titers to type 1a BVDV of > or = 1:32. The high prevalence of seropositive mule deer and isolation of BVDV suggests that this virus circulates in the mule deer population. The isolate described in this report is the second reported BVDV isolate from free-ranging deer in North America and the first from a mule deer. PMID:11310881
Van Campen, H; Ridpath, J; Williams, E; Cavender, J; Edwards, J; Smith, S; Sawyer, H
A cross-sectional study was conducted from November 2011 to April 2012 in Chifra district of Afar and in Jigjiga Zone of Somali Regional States of Ethiopia with the aims of assessing the epidemiology of camelpox and isolate and molecularly characterize the virus. The study included a questionnaire, active disease search and virusisolation and sequencing. A total of 24 (4.50%) and 12 (3.0%) camels in Afar and Jigjiga respectively were found clinically sick of camelpox during the study period. The questionnaire survey indicated that camelpox is the most common disease in the areas in which 125 (96%) of the respondents reported the frequent occurrence of camelpox in their herds especially during rainy season. The PCR result revealed 12 out of 17 tested samples were positive, of which seven of them collected from Jigjiga zone showed the characteristic PCR positive bands of 881 bp size fragments while five of the Afar samples gave two faint bands. Ethiopian isolates, specially isolated from Somali have very high identity with comparable sequences of CMLV M-96 from Kazakhstan and CMLV CMS from Iran. Out of the total of 780 bp analogous sequences, Ethiopian isolates differ only in two positions, while CMLV-Teheran differed at four nucleotide positions. The successfull isolation and molecular characterization of camelpox virus in Ethiopia, which could help for early diagnosis and control of the disease in the country. PMID:23578726
Protocols have been established for the preparation of large amounts of pure measles virus intracellular nucleocapsids. As a result, it has been possible to routinely achieve nucleocapsid RNA yields of approximately 200 micrograms (from approximately 5 X 10(8) infected cells). Electrophoretic analysis of this RNA under denaturing conditions revealed a single species whose mass was estimated at approximately 4.8 X 10(6) daltons. Electron microscopic assessment of nucleocapsid RNA contour lengths corroborated the electrophoretic size determination. Total nucleocapsid RNA was shown to contain both negative- and positive-stranded species distributed in a ratio of 2 to 3 genome polarity molecules for each antigenome RNA. Hybridization studies established that all of the virus-specified polyadenylated RNAs were encoded by the negative-stranded nucleocapsid RNA and, therefore, that this nucleocapsid RNA was the measles genome. Examination of the measles virus-specified, polyadenylated transcription products by HCHO-agarose gel electrophoresis revealed at least nine distinct RNA species (rather than the six predicted measles mRNAs). The significance of these observations is discussed. Images
We report the pathotyping of six Australian isolates of Marek's disease virus-1 (MDV1) isolated between 1992 and 2004 and association of virulence with meq gene polymorphism. Unvaccinated and herpesvirus of turkeys (HVT)-vaccinated specific pathogen free chickens were challenged at day 5 with 500 plaque forming units of Marek's disease virus. The isolates induced gross Marek's disease lesions in 53 to 94% of unvaccinated chickens, and HVT induced a protective index ranging from 38 to 100% by 56 days post challenge. This experiment provides evidence that current Australian isolates of MDV1 vary significantly in pathogenicity. However, there was no clear evidence that the most virulent recent isolates were more pathogenic than isolates from the 1980s or that any of the isolates belong to the highest pathotype category of very virulent plus. Evidence is presented that virulence can be predicted by measurements taken as early as 13 days post challenge. The meq gene sequences of five of the isolates used in the experiment were determined. When compared with the very virulent US isolate Md5, there was a 177 base-pair insertion and distinct point mutations in each of the five isolates. There were no individual mutations in the meq sequences that correlated with levels of virulence. However, amino acid alignment of the five Australian and 14 international isolates revealed that the number of repeat sequences of four prolines (PPPP repeats) in the meq gene (overall range 2 to 8) was strongly associated with virulence across all isolates, with the most pathogenic isolates having the fewest number of repeats. The results suggest that the presence of the 177 base-pair insertion alone is not an indicator of attenuation. Rather, the number of PPPP repeats, independent of the presence of the insertion, is a better indicator of pathogenicity. PMID:22515535
Renz, Katrin G; Cooke, Julie; Clarke, Nadeene; Cheetham, Brian F; Hussain, Zahid; Fakhrul Islam, A F M; Tannock, Gregory A; Walkden-Brown, Stephen W
Puumala (PUU) virus causes a form of hemorrhagic fever with renal syndrome (HFRS), called nephropathia epidemica (NE), in Europe. HFRS is characterized by an increased capillary permeability, which we hypothesize is caused by hyperactivation of the host immune system, especially cellular immune responses. To identify cytotoxic T lymphocytes (CTLs) specific for the PUU virus from NE patients, we have made
Masanori Terajima; Heather Lin Van Epps; Dexin Li; Anita M. Leporati; Sarah E. Juhlin; Jukka Mustonen; Antti Vaheri; Francis A. Ennis
West Nile virus (WNV) was isolated from a patient who developed encephalitis while undergoing treatment with CHOP (cyclophosphamide, hydroxydoxorubicin, vincristine [Oncovin], predisone) and rituximab for a non-Hodgkin B-cell lymphoma. Both standard reverse transcription–polymerase chain reaction (RT-PCR) and Taqman RT-PCR established the diagnosis of WNV infection from cerebrospinal fluid (CSF). Several whole blood samples and one serum sample underwent further testing. CSF and serum samples were negative for WNV antibody; however, all samples were positive by both RT-PCR assays. Infectious virus was recovered from a blood sample, and its identity was confirmed by using a WNV-specific immunofluorescence assay. The complete WNV genomes determined from CSF and from the virusisolate adapted from cell culture were the same. The results represent the first complete WNV genome sequence obtained directly from human CSF and the first time that infectious WNV has been recovered from a patient with encephalitis in North America.
A fatal human case of Duvenhage virus (DUVV) infection in a Dutch traveller who had returned from Kenya was reported in 2007. She exhibited classical symptoms of rabies encephalitis with distinct pathological findings. In the present study we describe the isolation and characterization of DUVV in vitro and its passage in BALB/c mice. The virus proved to be neuroinvasive in both juvenile and adult mice, resulting in about 50% lethality upon peripheral infection. Clinical signs in infected mice were those of classical rabies. However, the distribution of viral antigen expression in the brain differed from that of classical rabies virus infection and neither inclusion bodies nor neuronal necrosis were observed. This is the first study to describe the in vitro and in vivo isolation and characterization of DUVV.
Koraka, Penelope; Martina, Byron E. E.; Roose, Jouke M.; van Thiel, Pieter-Paul A. M.; van Amerongen, Geert; Kuiken, Thijs; Osterhaus, Albert D. M. E.
From 18 tobacco mosaic virusisolates from tomato, which could be divided into 3 pathogenicity groups, the buoyant density, the S value, the base composition, the amino acid composition, and the behaviour of their tryptic peptides in thin-layer chromatography were compared. There were no differences in buoyant densities and S values. With respect to the other characteristics only small individual
Eighteen strains of Rift Valley fever (RVF) virus collected over a period of 38 years and isolated from diverse localities in Africa and from various hosts (human, animal and arthropod) were investi- gated by RT-PCR followed by sequencing of the NSS protein coding region. This region was chosen to analyse variability because, in contrast to the N protein, the NSS
A. A. Sall; A. Zanotto; H. G. Zeller; J. P. Digoutte; Y. Thiongane; M. Bouloy
A new strain of canine distemper virus, CDV-PS, has been isolated from dogs in China, and its complete genome has been sequenced and analyzed. The phylogenetic analysis suggests that CDV-PS belongs to the Asia-1 cluster and has low identity to the vaccine strain. PMID:23682141
It is generally accepted that all primary isolates of feline leukemia virus (FeLV) contain a subgroup A virus (FeLV-A) that is essential for transmission. In contrast, FeLV-B is thought to arise de novo in the infected animal through RNA recombination events with endogenous FeLV transcripts, presumably through copackaging of RNA from endogenous FeLV and exogenous FeLV-A. Here, we report the complete genome sequences of two novel strains of FeLV-B (FeLV-2518 and FeLV-4314) that were isolated in the absence of FeLV-A. The env genes of these isolates have been characterized previously, and the 3' recombination sites have been identified. We describe herein the 5' recombination breakpoints of each virus. These breakpoints were found to be within the signal peptide of the env gene and the reverse transcriptase-coding region, respectively. This is the first report of a recombination site within the pol gene of an FeLV-B genome and the first genetic characterization of multiple independently arising FeLV-B isolates that have been identified without a functional FeLV-A ancestral virus. PMID:23405366
A new strain of canine distemper virus, HLJ1-06, has been isolated from foxes in China, and its complete genome has been sequenced and analyzed. The phylogenetic analysis suggests that HLJ1-06 belongs to the Asia-1 cluster and has low identity to the vaccine strain.
A strain of encephalomyocarditis virus (EMCV), strain FJ13, has been isolated from South China tigers in China, and its complete genome has been sequenced and analyzed. Phylogenetic analysis suggests that FJ13 belongs to the EMCV-1 serotype, and it is highly prevalent in China. PMID:23990575
Infectious bronchitis virus (IBV) causes tremendous economic losses to the poultry industry. Here, we report the complete genome analysis results for a new natural recombination nephropathogenic IBV strain named SAIBK, which was isolated in the Sichuan province of China in 2005.
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...
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
Ahmed S Abdel-Moneim; Ahmad E Abdel-Ghany; Salama AS Shany
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...
The complete genome sequence of a Mexican West Nile virusisolate, TM171-03, included 46 nucleotide (0.42%) and 4 amino acid (0.11%) differences from the NY99 prototype. Mouse virulence differences between plaque-purified variants of TM171-03 with mutations at the E protein glycosylation motif suggest the emergence of an attenuating mutation.
Davis, C. Todd; Estrada-Franco, Jose; Navarro-Lopez, Roberto; Campomanes-Cortes, Arturo; Tesh, Robert B.; Weaver, Scott C.; Barrett, Alan D.T.
Classical swine fever (CSF), a highly contagious viral disease of pigs, is endemic in India. As there is no information concerning the accurate genetic typing of classical swine fever virus (CSFV) isolates in India, 16 CSF virusesisolated during 2005-2007 from domestic pigs in different districts of Assam were typed in 5' UTR (150 nucleotides). To confirm the genetic typing results and to study the genetic variability, selected viruses were also analyzed in E2 (190 nt) and NS5B gene (409 nt) regions. Phylogenetic analysis revealed that all the 16 CSFV isolates analyzed belonged to group 1 and subgroup 1.1 in contrast to the situation in other Asian countries. Additionally, analysis in E2 and NS5B region placed the Indian isolates in a clearly separated clade within subgroup 1.1. The results suggest that subgroup 1.1 CSF viruses are currently circulating in India, which is important for epidemiology and control of CSF. PMID:19896713
Four influenza type B virusesisolated in Russia during periods of relatively low (1987-8) or high (1990-1) influenza B activity were characterized antigenically using a microneutralization assay. These isolates were antigenically similar to contemporary reference strains from either of two separate lineages represented by B/Victoria/2/87 and B/Yamagata/16/88. The evolutionary relationships of the variable portion of the haemagglutinin (HA1) genes of these viruses were determined by comparison with influenza B HA1 sequences previously obtained. The Isolate B/USSR/2/87, collected during the 1987-8 influenza season, was found to be closely related to viruses on the B/Victoria/2/87 lineage that circulated during the 1988-9 influenza season in the United States. Sequence analysis of the isolates from the 1990-1 influenza season demonstrated cocirculation of viruses from both the B/Victoria/2/87 and B/Yamagata/16/88 lineages in Russia, confirming the antigenic analysis.
Hemphill, M. L.; Rota, P. A.; Ivanova, V. T.; Slepushkin, A. N.; Kendal, A. P.
Objectives: Mosquito densonucleosis viruses (DNVs) are known to persistently infect the insect cell line and mosquito population in nature, causing mortality in mosquitoes. Here we report the isolation and characterization of a DNV from Aedes aegypti and its distribution among different Ae. aegypti populations from India. Methods: We screened Ae. aegypti mosquito populations from different states of India by PCR.
A. Sivaram; P. V. Barde; S. R. P. Kumar; P. Yadav; M. D. Gokhale; A. Basu; D. T. Mourya
In 2000, three Newcastle disease virus (NDV) strains were isolated from outbreaks of infection in layers, ducklings, and geese in the same region of China during the same time period. Here, we report their complete genome sequences, which belong to the NDV genotype VIId. This discovery might provide clues as to the evolution of the NDVs of different avian origins. PMID:23950112
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...
The nucleotide sequences of the genomes of eleven molecular clones for non-subtype B isolates of human immunodeficiency virus type 1 are disclosed. The invention relates to the nucleic acids and peptides encoded by and/or derived from these sequences and ...
Since October 2010, an outbreak of porcine epidemic diarrhea (PED) has been observed in some provinces of China. Here we report the complete genome sequence of porcine epidemic diarrhea virus (PEDV) strain LC, which was recently isolated from sucking piglets that suffered from severe watery diarrhea in Guangdong. It will help in understanding the epidemiological and molecular characteristics of PEDV in China. PMID:23087112
In order to enhance the outcome of high-quality reverse transcriptase enzyme, an efficient biotechnology was developed of accumulating and isolating the avian myeloblastosis virus (AMV) in high titres from blood plasma of leukosis-free chickens. When commercial chickens are infected in most sensitive one-day age, the virus titre does not exceed the value of 10(12) particles per 1 ml of plasma. We used 3-4-day old leukosis free chickens and achieved a stable average titre of the virus of 5.10(12) particles/ml due to adaptation of the virus to such chickens and their selection for a high sensitivity to AMV. PMID:1430580
Rabies virus (RABV) causes severe neurological disease and death. As an important mechanism for generating genetic diversity in viruses, homologous recombination can lead to the emergence of novel virus strains with increased virulence and changed host tropism. However, it is still unclear whether recombination plays a role in the evolution of RABV. In this study, we isolated and sequenced four circulating RABV strains in China. Phylogenetic analyses identified a novel lineage of hybrid origin that comprises two different strains, J and CQ92. Analyses revealed that the virus 3? untranslated region (UTR) and part of the N gene (approximate 500 nt in length) were likely derived from Chinese lineage I while the other part of the genomic sequence was homologous to Chinese lineage II. Our findings reveal that homologous recombination can occur naturally in the field and shape the genetic structure of RABV populations.
In the present study, a total of 158 fecal samples were collected from diarrheal dogs younger than 1 year old in pet clinic in China. 20 specimens (20/158, 13%) were positive for Torque teno canis virus DNA using detection PCR. One representative positive isolate designated LDL was randomly selected, cloned and sequenced. The complete genome of the LDL Chinese strain was 2799 nucleotides in length and contains three open reading frames (ORFs), which encode 576 (ORF1), 101 (ORF2), and 243 (ORF3) aa. Compared with the human and other animal TTV genomes, the genome of the LDL strain is clearly smaller and shares 95% identity with Japanese cf-TTV10 strain (AB076002). Phylogenetic analysis showed that the present Chinese Torque teno canis virus LDL strain was also closely clustered with the previous Japanese cf-TTV10 strain, and formed a different branch together with Torque teno sus viruses 1 and 2 compared with other Torque teno viruses, Torque teno mini virus, and Torque teno midi virus. Our study demonstrated that Torque teno canis virus is present in China. PMID:21645561
A small focus of hemorrhagic fever (HF) cases occurred near Cochabamba, Bolivia, in December 2003 and January 2004. Specimens were available from only one fatal case, which had a clinical course that included fever, headache, arthralgia, myalgia, and vomiting with subsequent deterioration and multiple hemorrhagic signs. A non-cytopathic virus was isolated from two of the patient serum samples, and identified as an arenavirus by IFA staining with a rabbit polyvalent antiserum raised against South American arenaviruses known to be associated with HF (Guanarito, Machupo, and Sabiá). RT-PCR analysis and subsequent analysis of the complete virus S and L RNA segment sequences identified the virus as a member of the New World Clade B arenaviruses, which includes all the pathogenic South American arenaviruses. The virus was shown to be most closely related to Sabiá virus, but with 26% and 30% nucleotide difference in the S and L segments, and 26%, 28%, 15% and 22% amino acid differences for the L, Z, N, and GP proteins, respectively, indicating the virus represents a newly discovered arenavirus, for which we propose the name Chapare virus. In conclusion, two different arenaviruses, Machupo and Chapare, can be associated with severe HF cases in Bolivia.
Delgado, Simon; Erickson, Bobbie R.; Agudo, Roberto; Blair, Patrick J.; Vallejo, Efrain; Albarino, Cesar G.; Vargas, Jorge; Comer, James A.; Rollin, Pierre E.; Ksiazek, Thomas G.; Olson, James G.; Nichol, Stuart T.
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 virusisolate from the original Malaysian outbreak (NiV-M). To investigate potential differences between NiV-M and a Nipah virusisolate 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
DeBuysscher, Blair L; de Wit, Emmie; Munster, Vincent J; Scott, Dana; Feldmann, Heinz; Prescott, Joseph
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 virusisolate from the original Malaysian outbreak (NiV-M). To investigate potential differences between NiV-M and a Nipah virusisolate 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.
DeBuysscher, Blair L.; de Wit, Emmie; Munster, Vincent J.; Scott, Dana; Feldmann, Heinz; Prescott, Joseph
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 tristezavirus (CTV) and Xylella fastidiosa, causal agent for citrus variegated chlorosis...
Classical swine fever is a disease that is devastating the pig industry worldwide. Here, we report the complete genome sequences of two classical swine virus strains (YC11WB and PC11WB), isolated from Korean wild boars in 2011. Both strains belong to subgenotype 2.1b. The complete genome sequences of PC11WB and YC11WB are more similar to that of strain ZJ0801 (isolated in China) than to that of the SW03 strain isolated from domestic pigs in South Korea. PMID:23599291
Jeoung, Hye-Young; Lim, Ji-Ae; Lim, Seong-In; Kim, Jae-Jo; Song, Jae-Young; Hyun, Bang-Hun; Kim, Yong Kwan; An, Dong-Jun
Classical swine fever is a disease that is devastating the pig industry worldwide. Here, we report the complete genome sequences of two classical swine virus strains (YC11WB and PC11WB), isolated from Korean wild boars in 2011. Both strains belong to subgenotype 2.1b. The complete genome sequences of PC11WB and YC11WB are more similar to that of strain ZJ0801 (isolated in China) than to that of the SW03 strain isolated from domestic pigs in South Korea.
Jeoung, Hye-Young; Lim, Ji-Ae; Lim, Seong-in; Kim, Jae-Jo; Song, Jae-Young; Hyun, Bang-Hun; Kim, Yong Kwan
Sequence analysis of the fusion (F) gene of eight Malaysian NDV isolates showed that all the isolates were categorized as\\u000a velogenic viruses, with the F cleavage site motif 112R-R-Q-K-R116 or 112R-R-R-K-R116 at the C-terminus of the F2 protein and phenylalanine (F) at residue 117 at the N-terminus of the F1 protein. Phylogenetic analysis revealed that all of the isolates were
Sheau Wei Tan; Aini Ideris; Abdul R. Omar; Khatijah Yusoff; Mohd Hair-Bejo
Subgroup J avian leukosis virus (ALV-J) isolate GDKP1202 was isolated from a 50-day-old local yellow commercial broiler in the Guangdong province of China in 2012. Here we report the complete genomic sequence of the GDKP1202 isolate, which caused high mortality, serious growth suppression, thymic atrophy, and liver enlargement in commercial broilers. A novel potential binding site (5?-GGCACCTCC-3?) for c-myb was identified in the GDKP1202 genome. These findings will provide additional insights into the molecular characteristics in the genomes and pathogenicity of ALV-J.
We report the isolation, purification, genome-sequencing and characterization of a picorna-like virus from dead bees in Israel. Sequence analysis indicated that IAPV (Israeli acute paralysis virus) is a distinct dicistrovirus. It is most homologous to Kashmir bee virus and acute bee paralysis virus. The virus carries a 9487 nt RNA genome in positive orientation, with two open reading frames separated
Eyal Maori; Shai Lavi; Rita Mozes-Koch; Yulia Gantman; Yuval Peretz; Orit Edelbaum; Edna Tanne; Ilan Sela
Ten influenza virusisolates were obtained from infected pigs from different places in Shandong province showing clinical symptoms from October 2002 to January 2003. All 10 isolates were identified in China's National Influenza Research Center as influenza A virus of H9N2 subtype. The complete genome of one isolate, designated A\\/Swine\\/Shandong\\/1\\/2003(H9N2), was sequenced and compared with sequences available in GenBank. The results
Two Hungarian virusisolates from sweet pepper (K8) and melon (S4) were identified as cucumber mosaic virus (CMV) on the basis of host plant reactions and serology. The isolates were purified and antisera prepared. Homologous antiserum titers in double-diffusion tests were 256 (K8) and 512 (S4). They were serologically closely related to each other and to other CMV isolates. On
Highly pathogenic avian H5N1 influenza A viruses have spread throughout Asia since 2003. These viruses are highly lethal to birds and humans. Of the 74 confirmed human cases, 49 were fatal (as of Mar 30, 2005), raising concerns of a possible pandemic by these viruses. Despite the well-established pathogenicity of these viruses, the molecular mechanism for expressing such high virulence remains elusive. Thus, we examined the pathogenicity of the H5N1 virusesisolated in Vietnam in 2003-2004 using animal models (mouse, duck, and ferret). Viruses from humans were generally more pathogenic in mice and ferrets than those from birds. Indeed, one human isolate was even lethal to ferrets. The human isolate possessing Lys at amino acid position 627 of PB2 was more virulent than that possessing Glu at this position, underscoring the importance of Lys at this position 627 of PB2 for efficient growth in mammals. PMID:16308530
Background Human respiratory syncytial virus (RSV) causes severe respiratory disease in infants. Airway epithelial cells are the principle targets of RSV infection. However, the mechanisms by which it causes disease are poorly understood. Most RSV pathogenesis data are derived using laboratory-adapted prototypic strains. We hypothesized that such strains may be poorly representative of recent clinical isolates in terms of virus/host interactions in primary human bronchial epithelial cells (PBECs). Methods To address this hypothesis, we isolated three RSV strains from infants hospitalized with bronchiolitis and compared them with the prototypic RSV A2 in terms of cytopathology, virus growth kinetics and chemokine secretion in infected PBEC monolayers. Results RSV A2 rapidly obliterated the PBECs, whereas the clinical isolates caused much less cytopathology. Concomitantly, RSV A2 also grew faster and to higher titers in PBECs. Furthermore, dramatically increased secretion of IP-10 and RANTES was evident following A2 infection compared with the clinical isolates. Conclusions The prototypic RSV strain A2 is poorly representative of recent clinical isolates in terms of cytopathogenicity, viral growth kinetics and pro-inflammatory responses induced following infection of PBEC monolayers. Thus, the choice of RSV strain may have important implications for future RSV pathogenesis studies.
Since 1998, H3N2 viruses have caused epizootics of respiratory disease in pigs throughout the major swine production regions of the U.S. These outbreaks are remarkable because swine influenza in North America had previously been caused almost exclusively by H1N1 viruses. We sequenced the full-length protein coding regions of all eight RNA segments from four H3N2 viruses that we isolated from
Alexander I. Karasin; Melissa M. Schutten; Lynn A. Cooper; Catherine B. Smith; Kanta Subbarao; Gary A. Anderson; Suzanne Carman; Christopher W. Olsen
The virusisolation-immunoperoxidase test (ipx) on cell cultures and the reverse transcription-polymerase chain reaction (rt-pcr) assay were compared for the detection of bovine viral diarrhoea virus (bvdv) directly in serum samples. Material for this study consisted of 403 sera originating from cattle in 41 bvdv-infected Finnish dairy herds and one suckler cow herd. The presence of virus was demonstrated in
U. I Laamanen; E. P Neuvonen; E. M Yliviuhkola; P. M.-L Veijalainen
The influenza epidemic occurring in Minsk in the autumn of 1986 was caused by at least two drift variants of influenza A virus (H1N1 subtype) differing in two antigenic determinants of hemagglutinin from the virusesisolated in previous years (A/USSR/90/77, A/Dunedin/6/83). One of the epidemic drift variants was found to be identical in the antigenic properties of hemagglutinin with the reference A/Taiwan/1/86 virus. PMID:2480024
We studied the effect of entry inhibitors on 58 virusisolates derived during acute and chronic infection to validate these inhibitors in vitro and to probe whether viruses at early and chronic disease stages exhibit general differences in the interaction with entry receptors. We included members of all types of inhibitors currently identified: (i) agents that block gp120 binding to CD4 (CD4-IgG2 and monoclonal antibody [MAb] IgG1b12), (ii) compounds that block the interaction with CCR5 (the chemokine RANTES/CCL5, the small-molecule inhibitor AD101, and the anti-CCR5 antibody PRO 140), (iii) the fusion inhibitor enfuvirtide (T-20), and (iv) neutralizing antibodies directed against gp120 (MAb 2G12) and gp41 (MAbs 2F5 and 4E10). No differences between viruses from acute and chronic infections in the susceptibility to inhibitors targeting the CD4 binding site, CCR5, or fusion or to MAb 2G12 were apparent, rendering treatment with entry inhibitors feasible across disease stages. The notable exceptions were antibodies 2F5 and 4E10, which were more potent in inhibiting viruses from acute infection (P = 0.0088 and 0.0005, respectively), although epitopes of these MAbs were equally well preserved in both groups. Activities of these MAbs correlated significantly with each other, suggesting that common features of the viral envelope modulate their potencies.
H5N1 highly pathogenic avian influenza (HPAI) viruses have seriously affected the Asian poultry industry since their recurrence in 2003. While surveillance in southern China has revealed that H5N1 viruses underwent extensive genetic reassortment to generate many different viral genotype viruses, little is known concerning the genotypes of H5N1 virus that circulated in central China in recent years. In this study, 16 H5N1 influenza viruses were isolated from the poultry market in central China during late 2006 and early 2007, and the genotypes and pathogenicity of the viruses were identified and characterized. All eight segments of each virus were sequenced, and phylogenetic analysis showed that the two surface glycoprotein genes, hemagglutinin (HA) and neuraminidase (NA), of all the viruses were closely related to the H5N1 virusesisolated in poultry in southern China since 2006. Phylogenetic analysis of the internal protein genes indicated that four viral genotypes circulated in poultry markets in central China. The virulence of 7 of the 16 isolates was tested in chickens and mice. The results showed that the 7 isolates were highly pathogenic for SPF chickens, and had a varied virulence in mice. Our results indicate that the H5N1 viruses circulated in central China have diversified characteristics of genotype and virulence. PMID:19720095
Brevidensoviruses have an encapsidated, single-stranded DNA genome that predominantly has a negative polarity. In recent years, they have received particular attention due to their potential role in the biological control of pathogenic arboviruses and to their unnoticed presence in cell cultures as contaminants. In addition, brevidensoviruses may also be useful as viral vectors. This study describes the first genetic and biological characterization of a mosquito densovirus that was isolated in Brazil; moreover, we examined the phylogenetic relationship between this isolate and the other brevidensoviruses. We further demonstrate that this densovirus has the potential to be used to biologically control dengue virus (DENV) infection with in vitro co-infection experiments. The present study provides evidence that this densovirus isolate is a fast-spreading virus that affects cell growth and DENV infection. PMID:21655815
Mosimann, Ana Luiza Pamplona; Bordignon, Juliano; Mazzarotto, Giovanny Camacho Antevęre; Motta, Maria Cristina M; Hoffmann, Federico; Santos, Claudia Nunes Duarte Dos
Dengue virus infection has been recognized as an important public health problem in the Dominican Republic in the last decade. Complete genomic sequences of three strains of dengue type 2 (DEN-2) virus, DR23/01 and DR31/01 isolated from dengue fever (DF) patients, and DR59/01 isolated from a dengue hemorrhagic fever (DHF) patient, all with primary infection, in the Dominican Republic in 2001, have been established. This achievement constitutes the first genomic characterization of DEN-2 strains from the Dominican Republic. No amino acid differences were observed between the strains isolated from DHF and DF patients. They exhibited extensive homology with the strain from La Martinique, French West Indies. Although phylogenetic analysis was suggestive of their Southeast Asiatic origin, Dominican Republic strains and other Caribbean strains from La Martinique and Jamaica showed 26 amino acid changes that differed from both the Southeast Asia and native American genotypes. PMID:15284482
The nucleotide sequences of the genomic RNAs of Cucumber green mottle mosaic virus Korean watermelon isolate (CGMMV-KW) and Korean oriental melon isolate (CGMMV-KOM) were determined and compared to the sequences of other tobamoviruses including CGMMV strains W and SH. Each CGMMV isolate had a genome of 6,424 nucleotides. Each also had 60 and 176 nucleotides of 5' and 3' untranslated regions (UTRs), respectively, and four open reading frames (ORF1-4). ORFs 1 to 4 encode proteins of 129, 186, 29, and 17.4 kDa, respectively. The nucleotide and deduced amino acid sequences of CGMMV-KOM and CGMMV-KW were more than 98.3% identical. When compared to other CGMMV strains in a phylogenetic analysis they were found to form a distinct virus clade, and were more distantly related to other tobamoviruses (23.5-56.7% identity). PMID:14744034
Kim, Sang-Min; Lee, Jung-Myung; Yim, Kyu-Ock; Oh, Man-Ho; Park, Jin-Woo; Kim, Kook-Hyung
The paralytic potential of the poliovirus was recognized as early as the 14th century B.C. as illustrated in Egyptian art. But it is only after the four last decades that methods for their concentration from water and their identification were performed. Among several of them the adsorption-elution method was retained. Nevertheless two important barriers had to be ran-over. The first one was the concentration-elution steps on different materials which had to be improved. The second one was the typing method which had to move from particle by particle identification to entire viral population. Despite of these advances only a few cytopathogenic serotypes were found. The reverse transcriptase-polymerase chain reaction with its far more wide spectrum allows the fast and direct identification of viral nucleic acids (or their fragments) of almost all viruses, cytopathogenic or not. With this method elevated amounts of drinking water samples were found positive for several non cytopathogenic viruses. The sanitary significance of these results has still to be proved. PMID:11501259
The virulence of six pigeon-origin isolates of Newcastle disease virus (NDV) was evaluated before and after passage in white leghorn chickens. Four isolates were defined as pigeon paramyxovirus-1 (PPMV-1) and two isolates were classified as avian paramyxovirus-1 (APMV-1) with NDV monoclonal antibodies. The four PPMV-1 isolates were passaged four times in chickens, and the APMV-1 isolates were passaged only once. Infected birds were monitored clinically and euthanatized. Tissues were collected for histopathology, in situ hybridization with a NDV matrix gene digoxigenin-labeled riboprobe, and immunohistochemistry with an anti-peptide antibody to the nucleoprotein. Mean death time, intracerebral pathogenicity index, and intravenous pathogenicity index tests performed before and after passage in chickens demonstrated increased virulence of the passaged PPMV-1 isolates and high virulence of the original isolates of APMV-1. Sequence analysis of the fusion protein cleavage site of all six isolates demonstrated a sequence typical of the virulent pathotype. Although the pathotyping results indicated a virulence increase of all passaged PPMV-1 isolates, clinical disease was limited to depression and some nervous signs in only some of the 4-wk-old specific-pathogen-free white leghorns inoculated intraconjunctivally. However, an increased frequency of clinical signs and some mortality occurred in 2 wk olds inoculated intraconjunctivally with passaged virus. Histologically, prominent lesions in heart and brain were observed in birds among all four groups inoculated with the PPMV-1 isolates. The behavior of the two pigeon-origin APMV-1 isolates when inoculated into chickens was characteristic of velogenic viscerotropic NDVs and included necro-hemorrhagic lesions in the gastrointestinal tract. PMID:11785895
Sugarcane yellow leaf virus (SCYLV) (genus Polerovirus, family Luteoviridae), the causal agent of sugarcane yellow leaf disease (YLD), was first detected in China in 2006. To assess the distribution of SCYLV in the major sugarcane-growing Chinese provinces, leaf samples from 22 sugarcane clones (Saccharum spp. hybrid) showing YLD symptoms were collected and analyzed for infection by the virus using reverse transcription PCR (RT-PCR), quantitative RT-PCR, and immunological assays. A complete genomic sequence (5,879 nt) of the Chinese SCYLV isolate CHN-FJ1 and partial genomic sequences (2,915 nt) of 13 other Chinese SCYLV isolates from this study were amplified, cloned, and sequenced. The genomic sequence of the CHN-FJ1 isolate was found to share a high identity (98.4-99.1 %) with those of the Brazilian (BRA) genotype isolates and a low identity (86.5-86.9 %) with those of the CHN1 and Cuban (CUB) genotype isolates. The genetic diversity of these 14 Chinese SCYLV isolates was assessed along with that of 29 SCYLV isolates of worldwide origin reported in the GenBank database, based on the full or partial genomic sequence. Phylogenetic analysis demonstrated that all the 14 Chinese SCYLV isolates clustered into one large group with the BRA genotype and 12 other reported SCYLV isolates. In addition, five reported Chinese SCYLV isolates were grouped with the Peruvian (PER), CHN1 and CUB genotypes. We therefore speculated that at least four SCYLV genotypes, BRA, PER, CHN1, and CUB, are associated with YLD in China. Interestingly, a 39-nt deletion was detected in the sequence of the CHN-GD3 isolate, in the middle of the ORF1 region adjacent to the overlap between ORF1 and ORF2. This location is known to be one of the recombination breakpoints in the Luteoviridae family. PMID:22752564
Avian influenza H5N1 viruses pose a significant threat to human health because of their ability to infect humans directly. In the paper, three highly pathogenic H5N1 influenza viruses were isolated from three species of migratory birds in Qinghai Province of China in 2006. The analysis of the genome sequences indicated that the three isolates shared high homology with each other (94% to 99%). Three isolates shared a common ancestor and were closest to strains isolated from Qinghai and Siberia in 2005, but distinct from poultry viruses found in Southeast Asia. In experimental infection, all three viruses were highly pathogenic to chickens and mice. The results suggest that highly pathogenic avian influenza H5N1 viruses still exist in the migratory birds and could spread to other regions with wild bird migration. PMID:17626485
Virusisolates were obtained from three ramie samples (Boehmeria nivea L.) showing yellow mosaic symptoms collected in Jiangsu and Zhejiang provinces, China. Comparison of partial DNA-A fragments amplified with begomovirus universal primers PA/PB revealed that these viral isolates shared a high sequence identity. The complete DNA-A sequences of two isolates J4 and Z1 were determined to be 2736 and 2737 nts, respectively, sharing 94.7% nucleotide sequence identity with each other. Also, the DNA-B components were identified for J4 and Z1 isolates and comprised 2717 and 2719 nts, respectively, sharing 88.6% nucleotide sequence identity with each other. Furthermore, sequence alignment and phylogenetic analysis showed that J4 and Z1 isolates had the highest sequence identities (93.6-94.7%) with isolates of Ramie mosaic virus (RamMV) for DNA-A. These molecular data suggested that J4 and Z1 may be two different isolates of RamMV. This is the first report about the occurrence of a bipartite begomovirus in these regions of China. PMID:20822317
Hepatitis delta virus (HDV) is a subviral agent of hepatitis B virus (HBV), and its life cycle is dependent on HBV. It is commonly accepted that HDV has eight distinct genotypes. In this study, the complete nucleotide sequences of HDV genomes isolated from nine Turkish patients were obtained by RT-PCR using two pairs of primers that cover the entire HDV genome. PCR products were sequenced directly. The results showed that these 9 isolates were approximately 1680 base pairs in length and clustered in the genotype HDV-1 branch when phylogenetic analysis was done with the sequences together with the complete sequences of HDV genomes representing each genotype retrieved from GenBank. Analysis of a portion of the large hepatitis D antigen (L-HDAg) gene showed that sequence similarity among these Turkish isolates is between 87.4 and 97.1%, and the Turkish isolates have the most sequence similarity to HDV-1 (90.5%), while they have the least sequence similarity to HDV-3 (64.1%). Full-genome analysis indicates that the sequence similarity is between 80.7 and 95.4%, and the highest sequence similarity is 84.8% (between the Turkish isolates and HDV-1). The lowest sequence similarity is 56.4% (between the Turkish isolates and HDV-3). In conclusion, phylogenetic analysis shows that the Turkish HDV isolates belong to HDV-1. PMID:21984217
The authors describe the isolation and identification of orf virus (ORFV) from an outbreak in a flock of sheep at Mukteswar, Uttarakhand, India, in 2009. The causative agent, ORFV was successfully isolated in primary lamb testes cells and identified using a semi-nested diagnostic polymerase chain reaction (PCR) and sequence and phylogenetic analyses of immunogenic envelope protein (B2L) coding gene. The affected animals showed characteristic proliferative skin lesions around the mouth and on nostrils and, in a few animals, lesions were also noticed on the tongue irrespective of age and sex. The morbidity, mortality and case fatality rates observed were 6%, 45% and 13%, respectively. Clinical samples were initially screened by counter immuno-electrophoresis and the serum neutralisation test; further positive skin scabs were tested with diagnostic PCR and virusisolation was performed on primary or secondary lamb testes cultures. Sequencing and phylogenetic analyses of the sheep isolate based on the B2L gene revealed that the isolate was closest to a goat isolate retrieved from an outbreak at the same geographic location in 2000. Furthermore, it also showed close genetic similarities with other Indian isolates reported earlier. Regular and systematic investigation of outbreaks is necessary to monitor the disease in susceptible populations. The development of rapid diagnostic methods as well as effective vaccine to control this infection not only from India but also other parts of the world is called for. PMID:21947970
Three new legume diseases in The Netherlands are described:Wisteria vein mosaic, pea necrosis, and pea leafroll mosaic. In particle size and morphology and in host reaction the virusisolates resembled bean yellow mosaic virus (BYMV), but they were readily distinguishable in several test plants.
Tomato spotted wilt virus (TSWV.Tospovirus) was detected in tomatillo (Physalis ixocarpa) plants, with symptoms of leaf spots, wilting, and generalized necrosis, grown in the states of México, Puebla and Morelos. Symptoms' expression in susceptible hosts indicated the possible presence of strains of this virus. The isolates of TSWV were identified as belonging to the group of localized lesion pathogens (TSWV-PL),
Rodolfo De La Torre-Almaráz; Hobbs A. Houston; Rodrigo Valverde
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 virusesisolated 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 virusesisolated 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
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 virusesisolated 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 virusesisolated 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.
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.
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-isolatedviruses 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 virusesisolated 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.
Summary. ?The partial sequencing of the internal and the neuraminidase genes of isolate 268\\/96 obtained from a woman with conjunctivitis\\u000a showed all seven to have closest homology with avian influenza viruses. The entire nucleotide sequence of the haemagglutinin\\u000a gene of 268\\/96 had close, 98.2%, homology with an H7N7 virusisolated from turkeys in Ireland in 1995. This appears to be\\u000a the
Human T-cell leukemia (or lymphotropic) virus type II (HTLV-II) was isolated from eight HTLV-seropositive patients, six of whom were also infected with human immunodeficiency virus, by cocultivation of peripheral blood mononuclear cells (PBMCs) with BJAB, a continuous B-cell line. Restriction endonuclease mapping of the proviruses demonstrated consistent differences among isolates, and two distinct physical map patterns were observed. The results suggest the existence of two closely related molecular subtypes of HTLV-II, which are tentatively designated HTLV-IIa and HTLV-IIb. This finding was supported by preliminary nucleotide sequence analysis of the env gene region encoding the transmembrane glycoprotein gp21, which showed consistent differences between the two proposed virus subtypes. Exploitation of differences in restriction endonuclease sites allowed polymerase chain reaction amplification to detect and differentiate the two subtypes in fresh PBMCs of HTLV-seropositive intravenous drug abusers (IVDAs). The results of these studies confirm that HTLV-II infection is the prominent HTLV infection in seropositive IVDAs and also show that infection with both subtypes occurs. The finding of genetic heterogeneity in the HTLV-II group of viruses may have important implications for studies on its role in human disease and will be useful in characterizing the viruses present in newly discovered endemic foci in New World indigenous populations. Images
Hall, W W; Takahashi, H; Liu, C; Kaplan, M H; Scheewind, O; Ijichi, S; Nagashima, K; Gallo, R C
Summary Duck hepatitis was first reported in 1985 in Korea. The complete nucleotide sequence of two past Korean isolates, DHV-HS and\\u000a DHV-HSS, isolated in 1994 and 1995, and four recent Korean isolates, AP-03337, AP-04009, AP-04114 and AP-04203 isolated in\\u000a 2003 and 2004, were determined. Phylogenetic analysis using the 3D protein sequence confirmed that the previously characterized\\u000a duck hepatitis virus type 1
M.-C. Kim; Y.-K. Kwon; S.-J. Joh; S.-J. Kim; C. Tolf; J.-H. Kim; H.-W. Sung; A. M. Lindberg; J.-H. Kwon
Dengue is an emerging disease in Nepal and was first observed as an outbreak in nine lowland districts in 2006. In 2010, however, a large epidemic of dengue occurred with 4,529 suspected and 917 serologically-confirmed cases and five deaths reported in government hospitals in Nepal. The collection of demographic information was performed along with an entomological survey and clinical evaluation of the patients. A total of 280 serum samples were collected from suspected dengue patients. These samples were subjected to routine laboratory investigations and IgM-capture ELISA for dengue serological identification, and 160 acute serum samples were used for virusisolation, RT-PCR, sequencing and phylogenetic analysis. The results showed that affected patients were predominately adults, and that 10% of the cases were classified as dengue haemorrhagic fever/ dengue shock syndrome. The genetic characterization of dengue virusesisolated from patients in four major outbreak areas of Nepal suggests that the DENV-1 strain was responsible for the 2010 epidemic. Entomological studies identified Aedes aegypti in all epidemic areas. All viruses belonged to a monophyletic single clade which is phylogenetically close to Indian viruses. The dengue epidemic started in the lowlands and expanded to the highland areas. To our knowledge, this is the first dengue isolation and genetic characterization reported from Nepal.
The complete genomic sequence of a watermelon isolate of Cucumber green mottle mosaic virus (CGMMV-LN) in Liaoning province was determined and compared with other cucurbit-infecting tobamoviruses. The genomic RNA of CGMMV-LN comprised 6422 nt, and 5'- and 3'- noncoding regions consisted of 59 nt and 175 nt, respectively. The encoded four proteins were two replicase proteins of 186 kD and 129 kD, move protein of 29 kD and coat protein of 17.4 kD. The alignment results of complete nucleotide sequence showed that CGMMV-LN shared identities of 97.6%-99.3% with four other CGMMV isolates, but only shared identities of 61.7%-62.8% with three other tobamoviruses. Homology trees generated from replicase proteins of 186 kD and coat proteins suggested that cucurbit-infecting tobamoviruses could be separated into two subgroups: subgroup I comprising all the isolates of CGMMV and subgroup II comprising Cucumber fruit mottle mosaic virus, Kyuri green mottle mosaic virus and Zucchini green mottle mosaic virus. PMID:19437890
Dengue is an emerging disease in Nepal and was first observed as an outbreak in nine lowland districts in 2006. In 2010, however, a large epidemic of dengue occurred with 4,529 suspected and 917 serologically-confirmed cases and five deaths reported in government hospitals in Nepal. The collection of demographic information was performed along with an entomological survey and clinical evaluation of the patients. A total of 280 serum samples were collected from suspected dengue patients. These samples were subjected to routine laboratory investigations and IgM-capture ELISA for dengue serological identification, and 160 acute serum samples were used for virusisolation, RT-PCR, sequencing and phylogenetic analysis. The results showed that affected patients were predominately adults, and that 10% of the cases were classified as dengue haemorrhagic fever/ dengue shock syndrome. The genetic characterization of dengue virusesisolated from patients in four major outbreak areas of Nepal suggests that the DENV-1 strain was responsible for the 2010 epidemic. Entomological studies identified Aedes aegypti in all epidemic areas. All viruses belonged to a monophyletic single clade which is phylogenetically close to Indian viruses. The dengue epidemic started in the lowlands and expanded to the highland areas. To our knowledge, this is the first dengue isolation and genetic characterization reported from Nepal. PMID:24155651
Background Herpes simplex viruses exist as two major serotypes, type 1 (HSV-1) and type 2 (HSV-2). Determination of type, either HSV-1 or HSV-2, is important in accurate diagnosis and clinical control of transmission. Several tests are available for typing HSV, including a monoclonal antibody specific for glycoprotein G and several PCR assays. Findings A clinical isolate was identified as herpes simplex virus, but tested negative for both HSV-1 and HSV-2 antigens using type-specific monoclonal antibody assays. The isolate was determined to be HSV-1 by PCR analysis. A mutation which likely caused the monoclonal antibody non-reactivity was found in glycoprotein G. Phylogenetic analysis revealed two groups of HSV, one with the mutation and one without. Three population studies examining mutations in HSV-1 glycoprotein G were analyzed by chi-squared test. To this point, the epitope which the monoclonal antibody recognizes was only found in HSV-1 isolates from human European populations (p < 0.0001). Conclusions These findings suggest that the PCR-based methods for HSV typing may be more useful than the standard monoclonal antibody test in areas of the world where the variant in glycoprotein G is more prevalent.
Newly discovered TT virus (TTV) is widely distributed in human populations. To understand more about the relationship between TTV and its hosts, we tested 400 sera from various nonhuman primates for the presence of TTV DNA by PCR assay. We collected serum samples from 24 different species of nonhuman primates. TTV DNA was determined by PCR with primers designed from the 5?-end region of the TTV genome. Nucleotide sequencing and phylogenetic analysis of viral genomes were also performed. TTV DNA was detected in 87 of 98 (89%) chimpanzees and 3 of 21 (14%) crab-eating macaques. Nucleotide sequences of the PCR products obtained from both animals were 80 to 100% identical between two species. In contrast, the sequences differed from TTV isolates in humans by 24 to 33% at the nucleotide level and 36 to 50% at the amino acid level. Phylogenetic analysis demonstrated that all TTV isolates obtained from simians were distinct from the human TTV isolates. Furthermore, TTV in simians, but not in humans, was classified into three different genotypes. Our results indicate that TTV in simians represents a group different from, but closely related to, TTV in humans. From these results, we tentatively named this TTV simian TTV (s-TTV). The existence of the s-TTV will be important in determining the origin, nature, and transmission of human TTV and may provide useful animal models for studies of the infection and pathogenesis of this new DNA virus.
The nucleotide sequences of a specific region of the glycoprotein gene were compared among 63 strains of viral haemorrhagic septicaemia virus (VHSV) isolated from fish in France between 1971 and 1999. The analysis was performed on a region corresponding to amino acids 238 to 331 of the glycoprotein gene, also designated the V2 region and previously shown to accumulate most of the mutations. The sequences of many VHSV isolates were found to be identical or very conserved. An isolate, designated L59X, obtained from elver in the Loire estuary, depicted a higher degree of divergence compared to the other French isolates. The deduced amino-acid sequences were analysed together with the results of neutralisation tests performed using monoclonal antibody 168m4 specific to serotype 1. Non-neutralised VHSV strains had mutations in the region corresponding to the previously described 168m4 epitope. Phylogenetic analysis showed that all the VHSV isolates studied, except L59X, belong to genotype I, previously described as containing VHSV strains isolated from continental Europe. Most of the VHSV isolates studied were found to be genetically related to one of the previously described VHSV strains representative of the major serotypes. Isolate L59X, which was the only French marine strain studied, was found to belong to genotype II, previously shown to encompass the VHSV strains isolated from the British Isles coastal waters. Overall there was a good correlation between the geographical origin of the studied isolates and their genetic characteristics. PMID:12517003
Thiéry, R; de Boisséson, C; Jeffroy, J; Castric, J; de Kinkelin, P; Benmansour, A
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
Selvarajan, R; Mary Sheeba, M; Balasubramanian, V; Rajmohan, R; Dhevi, N Lakshmi; Sasireka, T
Encephalomyocarditis virus (EMCV) can infect many host species and cause acute myocarditis and sudden death in preweaned piglets. In this study, an EMCV strain (NJ08) was isolated from newborn pigs with clinical signs on a pig farm in mideastern China. It was identified by indirect immunofluorescence assay and reverse-transcription polymerase chain reaction. Experiments showed that the isolate could cause severe clinical symptoms and pathological changes in mice but no obvious clinical and pathological changes in commercial piglets. Complete genomic sequencing showed that the NJ08 strain was 78.3% to 100% identical with other isolates in regions coding for various proteins. Phylogenetic analysis showed that the NJ08 isolate belonged to subgroup Ia. This study confirmed that an EMCV isolate from pigs could be fatal to mice and provided new epidemiologic data on EMCV in China.
Rapid and sensitive detection is a key step in the effective and early response to the global hazard of various viral diseases. In this study, an integrated isolation of hepatitis B virus (HBV)-specific DNA fragment by magnetic nanoparticles (MNPs) and its immediate analysis by microchip CGE was performed. Microfluidic CE chip was used to accommodate the complete process of viral DNA isolation by MNPs including hybridization and thermal denaturation followed by CE separation. Beforehand, calibration curves of HBV fragments were constructed. For isolation by MNPs, specific streptavidin-biotin interaction was used to bind complementary HBV fragment to magnetic particles. After analysis of isolated HBV by regular MNPs method, innovative approach was performed. The commercial CE chip (Bio-rad) was successfully used to execute HBV fragment isolation. Detection using LIF with detection limit of 1 ng/mL was accomplished. PMID:23483558
Three strains of porcine reproductive and respiratory syndrome virus (PRRSV) were isolated in porcine lung macrophage (PLM) cultures from three swine herds. This has been the first successful isolation of PRRSV in the Czech Republic and the strains received the designations CAPM V-501, CAPM V-502 and CAPM V-503, respectively. All the three isolates in PLM were identified by immunofluorescence and immunoperoxidase tests and the strain CAPM V-502 also by electron microscopy using the ultrathin section technique. The strain CAPM V-502 has been adapted to the cell line MARC-145. Viral RNA in PLM cultures infected with any of the isolated PRRSV strains was demonstrated by RT-PCR targeted to the more conserved ORF 7 genomic region encoding the nucleocapsid protein. The assessment of PCR products in agarose gel revealed a uniform size of 394 bp in all the three isolates and the European prototype strain Lelystad used as positive control. PMID:9416008
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
Dahlia mosaic disease of the ornamental flowering plant Dahlia is caused by two caulimoviruses, dahlia mosaic virus (DMV) and dahlia common mosaic virus (DCMV). We used a rolling-circle amplification method to amplify, clone and determine for the first time the full genome sequence of a DCMV isolate from New Zealand (DCMV-NZ). Within the 7949-bp circular double-stranded retro-transcribing DCMV-NZ DNA, we identified six putative open reading frames, typical of all genomes in the family Caulimoviridae. The availability of the complete DCMV sequence provides a reference genome against which all others can be compared. PMID:21960043
Egg drop syndrome virus (EDSV) strain FJ12025 was isolated from a 9-day-old Muscovy duckling. The results of the sequence showed that the genome of strain FJ12025 is 33,213 bp in length, with a G+C content of 43.03%. When comparing the genome sequence of strain FJ12025 to that of laying duck original strain AV-127, we found 50 single-nucleotide polymorphisms (SNPs) between the two viral genome sequences. A genomic sequence comparison of FJ12025 and AV-127 will help to understand the phenotypic differences between the two viruses. PMID:23969050
To study the receptor specificities of H1 and H3 influenza virusesisolated recently from pigs, we employed the analogues of natural receptors, namely sialyloligosaccharides conjugated with polyacrylamide in biotinylated and label free forms. All Madin-Darby canine kidney (MDCK) cell-propagated viruses with human H3 or classical swine H1 hemagglutinins bound only to Neu5Ac?2–6Gal?1-bearing polymers, and not to Neu5Ac?2–3Gal?1-bearing polymers. This receptor-binding
Alexandra S. Gambaryan; Alexander I. Karasin; Alexander B. Tuzikov; Alexander A. Chinarev; Galina V. Pazynina; Nicolai V. Bovin; Mikhail N. Matrosovich; Christopher W. Olsen; Alexander I. Klimov
Venezuelan equine encephalitis virus (VEEV) is a prototypical enveloped ssRNA virus of the family Togaviridae. To better understand alphavirus assembly, we analyzed newly formed nucleocapsid particles (termed pre-viral nucleocapsids) isolated from infected cells. These particles were intermediates along the virus assembly pathway, and ultimately bind membrane-associated viral glycoproteins to bud as mature infectious virus. Purified pre-viral nucleocapsids were spherical with a unimodal diameter distribution. The structure of one class of pre-viral nucleocapsids was determined with single particle reconstruction of cryo-electron microscopy images. These studies showed that pre-viral nucleocapsids assembled into an icosahedral structure with a capsid stoichiometry similar to the mature nucleocapsid. However, the individual capsomers were organized significantly differently within the pre-viral and mature nucleocapsids. The pre-viral nucleocapsid structure implies that nucleocapsids are highly plastic and undergo glycoprotein and/or lipid-driven rearrangements during virus self-assembly. This mechanism of self-assembly may be general for other enveloped viruses.
Lamb, Kristen; Lokesh, G.L. [Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555-0647 (United States); Sherman, Michael [Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555-0647 (United States); W.M. Keck Center for Virus Imaging, University of Texas Medical Branch, Galveston, TX 77555-0647 (United States); Watowich, Stanley, E-mail: email@example.com [Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555-0647 (United States); W.M. Keck Center for Virus Imaging, University of Texas Medical Branch, Galveston, TX 77555-0647 (United States)
Japanese encephalitis virus (JEV) was isolated from the cerebrum of a calf which showed severe neurological symptoms in late September 2009, and the JEV isolate was revealed to be of genotype 1. This is the first report describing the isolation of genotype 1 JEV from cattle. PMID:23885004
Sacbrood virus (SBV) is one of the most destructive honey bee viruses. The virus causes failure to pupate and death in both larvae and adult bees. Genetic analysis of SBV infected honey bees (Apis cerana) from five different provinces was carried out based on three nucleotide sequences; one partial structural protein coding sequence and two non-structural protein coding sequences. Sequences amplified by three specific primer pairs were aligned and compared with reference sequences deposited in the GenBank database. Sequence alignments revealed a low level of sequence variation among Korean isolates (? 98.6% nucleotide identity), regardless of the genome regions studied or the geographic origins of the strains. Multiple sequence comparisons indicated that Korean SBV isolates are genetically closely related to Chinese and other Asian strains. Interestingly, the Korean SBV isolates showed a number of unique nucleotides and amino acids that had not been observed in other published strains. Korean and other Asian isolates from the host A. cerana and the UK, European and Japanese strains from the host Apis mellifera showed differences in nucleotide and deduced amino acid identities. This suggests that host-specificity exists among SBV strains isolated from different species. Phylogenetic relatedness between compared sequences was analyzed by MEGA 4.1 software using the neighbor-joining (NJ) method with a boot-strap value of 1000 replicates. Obtained topologies were in agreement with previous studies, in which a distinct group of SBV was formed by UK and European genotypes and another group was comprised of Asian genotypes including strains that originated from China, Japan (japonica), India and Nepal. However, phylogeny based on a partial protein structural coding sequence grouped all Korean SBV isolates identified in A. cerana as a separate cluster. Our findings suggest that further study, including Korean SBV isolated from A. mellifera, is needed. PMID:22221381
Three major outbreaks of avian influenza (AI) occurred in Thailand. During the third episode in October 2005, we have isolated H5N1 viruses from one human case and three poultry cases. The whole genomes of AI viruses from human, chickens and quail from the outbreaks were characterized. Sequence analysis of eight gene segments revealed that the 2005 H5N1 virusesisolated in October 2005 were closely related to those recovered from chicken, tiger(s) and human(s) in January and July 2004. In addition, the genetic changes of the AI isolates at the HA cleavage site have been observed. PMID:16935377
Avian influenza A viruses rarely infect humans; however, when human infection and subsequent human-to-human transmission occurs, worldwide outbreaks (pandemics) can result. The recent sporadic infections of humans in China with a previously unrecognized avian influenza A virus of the H7N9 subtype (A(H7N9)) have caused concern owing to the appreciable case fatality rate associated with these infections (more than 25%), potential instances of human-to-human transmission, and the lack of pre-existing immunity among humans to viruses of this subtype. Here we characterize two early human A(H7N9) isolates, A/Anhui/1/2013 (H7N9) and A/Shanghai/1/2013 (H7N9); hereafter referred to as Anhui/1 and Shanghai/1, respectively. In mice, Anhui/1 and Shanghai/1 were more pathogenic than a control avian H7N9 virus (A/duck/Gunma/466/2011 (H7N9); Dk/GM466) and a representative pandemic 2009 H1N1 virus (A/California/4/2009 (H1N1pdm09); CA04). Anhui/1, Shanghai/1 and Dk/GM466 replicated well in the nasal turbinates of ferrets. In nonhuman primates, Anhui/1 and Dk/GM466 replicated efficiently in the upper and lower respiratory tracts, whereas the replicative ability of conventional human influenza viruses is typically restricted to the upper respiratory tract of infected primates. By contrast, Anhui/1 did not replicate well in miniature pigs after intranasal inoculation. Critically, Anhui/1 transmitted through respiratory droplets in one of three pairs of ferrets. Glycan arrays showed that Anhui/1, Shanghai/1 and A/Hangzhou/1/2013 (H7N9) (a third human A(H7N9) virus tested in this assay) bind to human virus-type receptors, a property that may be critical for virus transmissibility in ferrets. Anhui/1 was found to be less sensitive in mice to neuraminidase inhibitors than a pandemic H1N1 2009 virus, although both viruses were equally susceptible to an experimental antiviral polymerase inhibitor. The robust replicative ability in mice, ferrets and nonhuman primates and the limited transmissibility in ferrets of Anhui/1 suggest that A(H7N9) viruses have pandemic potential. PMID:23842494
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.
Shin, Dongyoung; Richards, Stephanie L.; Alto, Barry W.; Bettinardi, David J.; Smartt, Chelsea T.
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
Shin, Dongyoung; Richards, Stephanie L; Alto, Barry W; Bettinardi, David J; Smartt, Chelsea T
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 isolatedvirus (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.
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 virusisolation 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 virusisolates were characterized as HPAI H5N1 virus by sequencing analysis. Phylogenetic comparisons revealed that the H5N1 virusesisolated 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 virusesisolated from poultry in Saudi Arabia in 2007-08. PMID:20521659
Ismail, Mahmoud Moussa; Khan, Owais Ahmed; Cattoli, Giovanni; Lu, Huaguang
This study has tested the effect of using homologous or heterologous equine influenza A virusisolates to evaluate serum antibody levels to influenza A virus in vaccinated and naturally-infected horses. In addition, the potential effect of antigenic selection of virus variants in egg versus tissue culture propagation systems was studied. Serum antibody levels in samples from horses recently infected with a local influenza A virusisolate (A/equine 2/Saskatoon/1/90) or recently vaccinated with a prototype isolate (A/equine 2/Miami/1/63) were assessed by hemagglutination inhibition and by single radial hemolysis using cell or egg-propagated A/equine 2/Saskatoon/1/90, A/equine 2/Miami/1/63 or A/equine 2/Fontainebleau/1/79. There were no significant differences in hemagglutination inhibition or single radial hemolysis antibody levels obtained with homologous or heterologous isolates or between viruses propagated in either eggs or cell culture. However there was a trend to higher titers in the hemagglutination inhibition assay when cell-propagated virus was used. These results suggest that antigenic variation in equine influenza A virusisolates and host-cell selection of antigenic variants during virus propagation may not be of sufficient magnitude to influence serological evaluation of antibody responses by hemagglutination inhibition or single radial hemolysis. PMID:8387870
Bogdan, J R; Morley, P S; Townsend, H G; Haines, D M
The first influenza pandemic of the 21st century was caused by novel H1N1 viruses that emerged in early 2009. Molecular evolutionary analyses of the 2009 pandemic influenza A H1N1 [A(H1N1)pdm09] virus revealed two major clusters, cluster I and cluster II. Although the pathogenicity of viruses belonging to cluster I, which became extinct by the end of 2009, has been examined in a nonhuman primate model, the pathogenic potential of viruses belonging to cluster II, which has spread more widely in the world, has not been studied in this animal model. Here, we characterized two Norwegian isolates belonging to cluster II, namely, A/Norway/3568/2009 (Norway3568) and A/Norway/3487-2/2009 (Norway3487), which caused distinct clinical symptoms, despite their genetic similarity. We observed more efficient replication in cultured cells and delayed virus clearance from ferret respiratory organs for Norway3487 virus, which was isolated from a severe case, compared with the efficiency of replication and time of clearance of Norway3568 virus, which was isolated from a mild case. Moreover, Norway3487 virus to some extent caused more severe lung damage in nonhuman primates than did Norway3568 virus. Our data suggest that the distinct replicative and pathogenic potentials of these two viruses may result from differences in their biological properties (e.g., the receptor-binding specificity of hemagglutinin and viral polymerase activity).
The naturally occurring Grapevine fanleaf virus (GFLV) recombinant isolate A17b was recovered from its grapevine host by sap inoculation and serial passages onto Gomphrena globosa, a pseudo local lesion herbaceous host, and Chenopodium quinoa, a systemic herbaceous host, to characterize some of its biological properties. Sequence analysis of the CP gene, in which a recombinational event was previously detected, demonstrated the genetic stability of recombinant isolate A17b over a 5-year period in its natural host as well as in C. quinoa. Also, recombinant isolate A17b was graft transmissible, as shown by an in vitro heterologous approach, and transmitted by the nematode Xiphinema index as readily as nonrecombinant GFLV isolates. Furthermore, despite a lower pathogenicity on Chenopodium amaranticolor, recombinant isolate A17b had a similar host range and induced similar symptoms in type and severity to nonrecombinant GFLV isolates. Interestingly, the use of infectious chimeric RNA2 transcripts in combination to RNA1 transcripts of GFLV strain F13 suggested no implication of the recombination event in the CP gene of isolate A17b in the reduced pathogenicity on C. amaranticolor. Altogether, recombinant isolate A17b had similar biological properties to GFLV nonrecombinant isolates. PMID:15968475
Since late 2010, porcine epidemic diarrhea virus (PEDV) has been re-emerging in immunized swine herds with devastating impact in the Hebei province of China. Seven prevailing strains of PEDV were isolated from fecal samples out of piglets suffering from severe diarrhea. The M gene of the seven PEDV isolates encompasses an open reading frame of 681 nucleotides, encoding a protein of 226 amino acids. The seven PEDV isolates showed 99.4-99.9 % nucleotide sequence identity and 98.2-99.1 % deduced amino acid identity. When compared with other Chinese isolates and foreign isolates, the seven isolates showed high nucleotide identity with the Thailand isolate M-NIAH1005 (99.6-99.9 %) and Korea isolate PFF188 (99.7-100 %), but low identity with other Chinese isolates (96.6-99.1 %) and with the vaccine strain CV777 used in China (97.8-98.2 %). Phylogenetic analyses showed that all seven Chinese field isolates were grouped together in the same cluster. Although CV777 was also separated into the same cluster with the seven isolates, they were belonged to different sub-cluster. These results showed that the seven prevailing isolates in China are closely related phylogenetically to each other and have close relationships with the Korean strain PFF188 and Thailand strain M_NIAH1005. However, they differ genetically from other Chinese isolates and the vaccine strain CV777. Therefore, a more efficient vaccine strain should be chosen to prevent outbreaks of PEDV in China. PMID:22585338
The establishment and characterization of a reticuloendotheliosis virus (strain T)-transformed lymphoblastoid cell line, designated TV-1, was reported. These cells, isolated from the spleen of a moribund chick infected with reticuloendotheliosis virus, were maintained in suspension culture for over 74 weeks at a stable generation time of 15 h. The cells were found to carry a female karyotype. Both TV-1 cells and cell-free TV-1 culture supernatant produced lesions and mortality patterns in chicks which were identical to those caused by reticuloendotheliosis virus (strain T) infection. Examination of TV-1 cells by electron microscope revealed the presence of C-type virions budding from the plasma membrane. Cytotoxicity assays and fluorescent antibody tests indicated the presence of B-cell determinants on the TV-1 cell surface membrane. Images
Recent investigations have shown the Egyptian fruit bat (Rousettus aegyptiacus) to be a natural reservoir for marburgviruses. To better understand the life cycle of these viruses in the natural host, a new reverse genetics system was developed for the reliable rescue of a Marburg virus (MARV) originally isolated directly from a R. aegyptiacus bat (371Bat). To develop this system, the exact terminal sequences were first determined by 5' and 3' RACE, followed by the cloning of viral proteins NP, VP35, VP30 and L into expression plasmids. Novel conditions were then developed to efficiently replicate virus mini-genomes followed by the construction of full-length genomic clones from which recombinant wild type and GFP-containing MARVs were rescued. Surprisingly, when these recombinant MARVs were propagated in primary human macrophages, a dramatic difference was found in their ability to grow and to elicit anti-viral cytokine responses. PMID:24074586
Albarińo, César G; Uebelhoer, Luke S; Vincent, Joel P; Khristova, Marina L; Chakrabarti, Ayan K; McElroy, Anita; Nichol, Stuart T; Towner, Jonathan S
La Crosse (LAC) virus, a California serogroup bunyavirus, is the leading cause of pediatric arboviral encephalitis in the United States and an emerging disease in Tennessee, West Virginia, and North Carolina. Human cases of LAC encephalitis in Tennessee and North Carolina have increased above endemic levels during 1997 to 1999 and may represent an expansion of a new southeastern endemic focus. This report describes the isolation of LAC virus from the exotic mosquito Aedes albopictus. The discovery of LAC virus in wild populations of Ae. albopictus coupled with its expanding distribution in the southeastern United States, suggests that this mosquito may become an important accessory vector, potentially increasing the number of human cases in endemic foci or expanding the range of the disease.
Gerhardt, R. R.; Gottfried, K. L.; Apperson, C. S.; Davis, B. S.; Erwin, P. C.; Smith, A. B.; Panella, N. A.; Powell, E. E.; Nasci, R. S.
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 virusesisolated 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 virusesisolated during 2007-08 necessitated that two of these strains be considered as potential H5N1 pre-pandemic vaccine candidates. PMID:20521654
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
Fourteen isolations of West Nile (WN) virus were obtained from four mosquito species (Culex pipiens , Cx. restuans , Cx. salinarius , and Culiseta melanura ) in statewide surveillance conducted from June through October 2000. Most isolates were obtained from mosquitoes collected in densely populated residential locales in Fairfield and New Haven counties, where the highest rates of dead crow sightings were reported and where WN virus was detected in 1999. Minimum field infection rates per 1,000 mosquitoes ranged from 0.5 to 1.8 (county based) and from 1.3 to 76.9 (site specific). Cx. restuans appears to be important in initiating WN virus transmission among birds in early summer; Cx. pipiens appears to play a greater role in amplifying virus later in the season. Cs. melanura could be important in the circulation of WN virus among birds in sylvan environments; Cx. salinarius is a suspected vector of WN virus to humans and horses.
Andreadis, T. G.; Anderson, J. F.; Vossbrinck, C. R.
The complete genome sequence of the KGH strain of the first human rabies virus, which was isolated from a skin biopsy of a patient with rabies, whose symptoms developed due to bites from a raccoon dog in 2001. The size of the KGH strain genome was determined to be 11,928 nucleotides (nt) with a leader sequence of 58 nt, nucleoprotein gene of 1,353 nt, phosphoprotein gene of 894 nt, matrix protein gene of 609 nt, glycoprotein gene of 1,575 nt, RNA-dependent RNA polymerase gene of 6,384 nt, and trailer region of 69 nt. Sequence similarity was compared with 39 fully sequenced rabies virus genomes currently available, and the result showed 70.6-91.6 % at the nucleotide level, and 82.8-97.9 % at the amino acid level. The deduced amino acids in the viral protein were compared with those of other rabies viruses, and various functional regions were investigated. As a result, we found that the KGH strain only had a unique amino acid substitution that was identified to be associated either with host immune response and pathogenicity in the N protein, or with a related region regulating STAT1 in the P protein, and related to pathogenicity in G protein. Based on phylogenetic analyses using the complete genome of 39 rabies viruses, the KGH strain was determined to be closely related with the NNV-RAB-H strain and transplant rabies virus serotype 1, which are Indian isolates, and was confirmed to belong to the Arctic-like 2 clade. The KGH strain was most closely related to the SKRRD0204HC and SKRRD0205HC strain when compared with Korean animal isolates, which was separated around the same time and place, and belonged to the Gangwon III subgroup. PMID:23242520
Park, Jun-Sun; Kim, Chi-Kyeong; Kim, Su Yeon; Ju, Young Ran
Complete nucleotide sequences of Tobacco leaf curl Japan virus (TbLCJV) isolates from infected tomato ( Lycopersicon esculentum) plants in Nara (-[Jp2], 2764 nt; -[Jp3], 2761 nt), Kochi (-[Koc], 2760 nt) and Yamaguchi (-[Yam], 2758 nt) Prefectures, of Japan were determined. These sequences were compared with each other and the sequences of further begomoviruses from Japan. TbLCJV, TbLCJV-[Jp2], TbLCJV-[Jp3], TbLCJV-[Koc], TbLCJV-[Yam], Honeysuckle yellow vein mosaic virus (HYVMV), Eupatorium yellow vein virus (EpYVV), EpYVV-[MNS2], EpYVV-[SOJ3], EpYVV-[Yam] and EpYVV-[Tob] are monophyletic. The intergenic region (IR) of TbLCJV has highest nucleotide sequence identity with that of HYVMV (93%) whereas the rest of the genomic DNA had higher identity with that of TbLCJV-[Jp2] or -[Jp3] (91 approximately 100%) than with that of HYVMV. In conclusion, TbLCJV has a chimeric genome which may have arisen by recombination between TbLCV-[Jp2] or -[Jp3]-like and HYVMV-like ancestors. Similarly, TbLCJV-[Yam] DNA has a hybrid genome, with a major parent HYVMV and minor parent TbLCJV-[Koc]. PMID:15168208
Sequence was determined for the coat protein (CP) gene and 3' non-translated region (3'NTR) of two vanilla mosaic virus (VanMV) isolates from Vanilla tahitensis, respectively from the Cook Islands (VanMV-CI) and French Polynesia (VanMV-FP). Both viruses displayed distinctive features in the N-terminal region of their CPs; for VanMV-CI, a 16-amino-acid deletion including the aphid transmission-related DAG motif, and for VanMV-FP, a stretch of GTN repeats that putatively belongs to the class of natively unfolded proteins. VanMV-FP CP also has a novel DVG motif in place of the DAG motif, and an uncommon Q//V protease cleavage site. The sequences were compared to a range of Dasheen mosaic virus (DsMV) strains and to potyviruses infecting orchids. Identity was low to DsMV strains across the entire CP coding region and across the 3'NTR, but high across the CP core and the CI-6K2-NIa region. In accordance with current ICTV criteria for species demarcation within the family Potyviridae, VanMV-CI and VanMV-FP are strains of DsMV that exclusively infect vanilla. PMID:16421638
Farreyrol, K; Pearson, M N; Grisoni, M; Cohen, D; Beck, D
In order to study the variation of low pathogenic avian influenza viruses (AIV) of H3 subtype in the natural reservoir, partial genetic characterisation of four AIV isolates of H3 subtype, recovered from wild mallards in Poland in 2006-2010, was performed. Phylogenetic analysis clearly confirms that there is a constant flow of AIV H3 between wild birds in Eurasia and Africa, and, to a limited degree, to North America (Alaska), with an occasional spill-over to poultry. The analysis of the PA gene of one isolate from 2010 suggests that it is closely related to several HPAI H5N1 viruses belonging to clade 2.3.2 and that, therefore, a reassortment event has occurred recently between low pathogenic and H5N1 highly pathogenic AIV. PMID:23921353
Immunological analysis performed by solid-phase enzyme immunoassay and complement fixation test revealed the presence of host cell antigenic components in purified intact influenza virions and isolated hemagglutinins and glycoproteins. Three antigens inherent to chick embryo cells were identified in virus preparations: the species-specific antigen, heterogenetic Forssman antigen, and one similar to human group A antigen. Antisera to purified whole-virion influenza vaccines, to glycoproteins and to hemagglutinin isolated from them were found to contain antibody populations which were conducive to broad cross reactions between them and heterotypic virus antigens. To avoid erroneous results, these antibody populations must be removed from the sera by pre-adsorption with host cell antigens. PMID:2461614
Rovnova, Z I; Isaeva, E I; Platonova, A L; Rogacheva, T A
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.
Control of drinking or bathing water quality in respect to viral contamination remains an unsolved problem. A highly sensitive isolation protocol was developed for concentration and detection of different enteric viruses from water samples. The three-step isolation procedure combines filtration with a positively charged nylon membrane, ultrafiltration and clean-up of the viral RNA with a silica based membrane. Detection of
Dengue is one of the most important diseases in the tropical and subtropical regions of the world, with an estimated 2.5 billion people being at risk. Detection of dengue virus infections has great importance for the clinical management of patients, surveillance, and clinical trial assessments. Traditionally, blood samples are collected in serum separator tubes, processed for serum, and then taken to the laboratory for analysis. The use of whole blood has the potential advantages of requiring less blood, providing quicker results, and perhaps providing better sensitivity during the acute phase of the disease. We compared the results obtained by reverse transcriptase PCR (RT-PCR) with blood drawn into tubes containing EDTA and those obtained by RT-PCR with blood samples in serum separator tubes from 108 individuals clinically suspected of being infected with dengue virus. We determined that the extraction of RNA from whole blood followed by RT-PCR resulted in a higher detection rate than the use of serum or plasma. Using a selection of these samples, we also found that our ability to detect virus by direct C6/36 cell culture and mosquito inoculation was enhanced by using whole blood but not to the same extent as that seen by the use of RT-PCR.
Klungthong, Chonticha; Gibbons, Robert V.; Thaisomboonsuk, Butsaya; Nisalak, Ananda; Kalayanarooj, Siripen; Thirawuth, Vipa; Nutkumhang, Naowayubol; Mammen, Mammen P.; Jarman, Richard G.
In Japan, between the end of December 2003 and March 2004, four outbreaks of acute, highly transmissible and lethal disease occurred in birds in three prefectures separated by 150-450 km, involving three chicken farms and a group of chickens raised as pets. The cause of each outbreak was an H5N1 influenza A virus-the first highly pathogenic virus to be isolated from the outbreaks in Japan since 1925. The H5N1 virus was also isolated from dead crows, apparently infected by contact with virus-contaminated material. These H5N1 viruses were antigenically similar to each other, but could be differentiated from other H5 viruses, including those isolated from Hong Kong in 1997 and 2003, by use of a panel of monoclonal antibodies in hemagglutination inhibition assays. Genetically, the H5N1 viruses in Japan were closely related to each other in all genes and were genetically closely related to a single isolate of genotype V that was isolated in 2003 in the Guandong Province of mainland China (A/chicken/Shantou/4231/2003). The virulence of the index isolate (A/chicken/Yamaguchi/7/2004) was studied in chickens and mice. Chickens intravenously or intranasally inoculated with the isolate died within 1 or 3 days of inoculation, respectively. In mice, although this virus replicated well in the lung without prior adaptation and spread to the brain, the dose lethal to 50% of the mice was 5 x 10(5) 50% egg infectious doses (EID50), which is less pathogenic than the Hong Kong 1997 H5N1 virusesisolated from humans. Our findings indicate that the H5N1 viruses associated with the influenza outbreaks in chickens in Japan were genotypically closely related to an H5N1 virusisolated from chicken in China in 2003 (genotype V), but were different from those prevalent in southeastern Asia in 2003-2004 (i.e., genotype Z) and that these highly pathogenic viruses can be transmitted to crows, which are highly susceptible to these viruses. PMID:15661149
Small polykaryocytes resulting from cell fusion are found in herpes simplex virus (HSV) lesions in patients, but their significance for viral spread and pathogenesis is unclear. Although syncytial variants causing extensive fusion in tissue culture can be readily isolated from laboratory strains, they are rarely found in clinical isolates, suggesting that extensive cell fusion may be deleterious in vivo. Syncytial mutations have previously been identified for several laboratory strains, but not for clinical isolates of HSV type 2. To address this deficiency, we studied a recent syncytial clinical isolate, finding it to be a mixture of two syncytial and one nonsyncytial strain. The two syncytial strains have novel mutations in glycoprotein B, and in vitro cell fusion assays confirmed that they are responsible for syncytium formation. This panel of clinical strains may be ideal for examining the effect of increased cell fusion on pathogenesis.
Muggeridge, Martin I. [Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, LA 71130 (United States) and Center for Molecular and Tumor Virology, Louisiana State University Health Sciences Center, Shreveport, LA 71130 (United States) and Feist-Weiller Cancer Center, Louisiana State University Health Sciences Center, Shreveport, LA 71130 (United States)]. E-mail: firstname.lastname@example.org; Grantham, Michael L. [Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, LA 71130 (United States); Center for Molecular and Tumor Virology, Louisiana State University Health Sciences Center, Shreveport, LA 71130 (United States); Johnson, F. Brent [Department of Microbiology and Molecular Biology, Brigham Young University, Provo, UT 84602 (United States)
Virus infection of Aspergillus foetidus was documented over 40 years ago and was one of the first mycovirus infections described in a filamentous fungus. The virus, named Aspergillus foetidus virus (AfV), contains at least two types of icosahedral particles, called AfV-fast (-F) and AfV-slow (-S) virions, based on their relative electrophoretic mobilities. Here, we report the complete nucleotide sequence of the AfV-F genome isolated from virions purified from the prototype isolate of the fungus. The AfV-F double-stranded (ds) RNA genome is tetra-segmented, and the plus strands of each of the four segments, but not the minus strands, are polyadenylated. The organisation and sequences of the four AfV-F dsRNAs are similar to those described for Alternaria alternata virus 1, which we propose is a member of an emerging mycovirus genus ("Alternavirus") and family ("Alternaviridae"), which also includes AfV-F. PMID:22760661
Kozlakidis, Zisis; Herrero, Noemi; Ozkan, Selin; Kanhayuwa, Lakkhana; Jamal, Atif; Bhatti, Muhammad F; Coutts, Robert H A
Reproduction of nodavirus disease was performed by experimental infection of sea bass eggs during fertilization or at larval stage 4 with 2 genetically distinguishable nodavirus strains (Sb1 and Sb2) isolated from sea bass collected along the Atlantic and Mediterranean French coast. The pathogenicity of the virus strains was assigned after detection of the virus by ELISA and immunohistochemistry (IHC). The Atlantic (Sb1) strain was more pathogenic than the Mediterranean (Sb2) strain during the fertilization step whilst both strains were pathogenic following experimental exposure of 4 d old larvae. Virus lesions developed in the brain 4 to 6 d following experimental exposure. Experimental ELISA proved very sensitive for detecting the nodavirus in Sb1 or Sb2 experimentally infected larvae, as well as in naturally infected sea bass larvae collected in French hatcheries or in barramundi larvae reared in the Pacific area. The development of an ELISA specific for the 2 nodavirus strains isolated from the sea bass should be useful for the detection of the virus, in addition to other techniques recommended by the Office International des Epizooties (OIE). PMID:11411641
A new polymerase chain reaction (PCR) assay for rapid diagnosis of contagious ecthyma was designed and applied to 21 clinical samples from Greece. This assay, which detects a highly conserved gene from the parapox genome, was evaluated for its sensitivity and specificity in order to be considered as a useful diagnostic tool. A comparative study with two published PCR protocols one using primers PPP1-PPP3, PPP1-PPP4 which targets putative virion envelope gene B2L and the other using VIR1-VIR2 primers which amplifies ORF virus interferon resistant (VIR) gene, as well as cell culture virus neutralization assay was carried out. All samples tested were amplified successfully with the PCR protocol established in the laboratory. The combination of primers PPP1-PPP3 and PPP1-PPP4 in a semi-nested PCR gave a positive result in 20 of 21 samples while primers VIR1-VIR2 failed to amplify successfully 7 of 21 samples. The diagnostic value of parapox viral DNA amplification was also compared with the results of virusisolation by cell culture and was positive in three samples that the virusisolation was obtained. PMID:16417927
Kottaridi, Christine; Nomikou, Kiki; Lelli, Rossella; Markoulatos, Panayotis; Mangana, Olga
Swine hepatitis E virus (HEV) is a newly identified potentially zoonotic agent that is possibly transmitted to humans from pigs. Swine HEV is prevalent in pig populations and does not cause abnormal clinical symptoms in infected pigs, further implicating a likelihood of a risk of transmission to humans by normal contact. To date in North America, only one strain of swine HEV (strain US swine) has been fully sequenced. In the present study, we identified a swine HEV isolate from pigs in Canada, designated the Arkell strain, and determined the full length of the genomic sequence. The genome of Canadian strain Arkell consisted of 7,242 nucleotides, excluding the poly(A) tail of at least 15 A residues. The genome contained three open reading frames (ORFs), ORF1, ORF2, and ORF3, which had coding capacities for proteins of 1,708, 660, and 122 amino acids, respectively. Comparative analysis of the full-length genomic sequence indicated that the sequence of strain Arkell was distinct from those of all other known HEV isolates by 13 to 27% and shared the highest degrees of identity with human HEV isolates US-1 and US-2, HEV isolate US swine, and the human and swine HEV isolates recently isolated in Japan. On the basis of sequence similarities and phylogenetic analyses, HEV strain Arkell was grouped into genotype 3. The sequence of the Arkell swine HEV isolate differed from those of HEV isolate US swine and HEV isolate Japan swine by 13 and 14%, respectively. To date, two isolates of swine HEV (isolates Arkell and SK3 [D. Yoo et al., Clin. Diagn. Lab. Immunol. 8:1213-1219, 2001]) have been identified in Canadian pigs, and their sequences also differ from each other by 11.8%. Our studies indicate that, as with human HEV strains, swine HEV isolates exhibit extensive genetic heterogeneity.
Epidemic outbreaks of Tomato yellow leaf curl virus (TYLCV) diseases occurred in greenhouse grown tomato (Solanum lycopersicum) plants of Busan (TYLCV-Bus), Boseong (TYLCV-Bos), Hwaseong (TYLCV-Hwas), Jeju Island (TYLCV-Jeju), and Nonsan (TYLCV-Nons) in Korea during 2008-2009. Tomato disease by TYLCV has never occurred in Korea before. We synthesized the full-length genomes of each TYLCV isolate from the tomato plants collected at each area and determined their nucleotides (nt) sequences and deduced the amino acids of six open reading frames in the genomes. TYLCV-Bus and -Bos genomes shared higher nt identities with four Japanese isolates -Ng, -Omu, -Mis, and -Miy. On the other hand, TYLCV-Hwas, -Jeju, and -Nons genomes shared higher nt identities with five Chinese isolates TYLCV-AH1, -ZJ3, -ZJHZ12, -SH2, -Sh10, and two Japanese isolates -Han and -Tosa. On the basis of a neighbor-joining tree, five Korean TYLCV isolates were separated into three clades. TYLCV-Bus and -Bos formed the first clade, clustering with four Japanese isolates TYLCV-Mis, -Omu, -Ng, and -Miy. TYLCV-Jeju and -Nons formed the second clade, clustering with two Chinese isolates -ZJHZ212 and -Sh10. TYLCV-Hwas was clustered with two Japanese isolates -Han and -Tosa and three Chinese isolates -AH1, -ZJ3, and -SH2. Two fragments that had a potentially recombinant origin were identified using the RDP, GENECONV, BootScan, MaxChi, Chimaera, SiScan, and 3Seq methods implemented in RDP3.41. On the basis of RDP analysis, all TYLCV isolates could originated from the interspecies recombination between TYLCV-Mld[PT] isolated from Portugal as a major parent and TYLCTHV-MM isolated from Myanmar as a minor parent. PMID:20963475
Kim, Sue Hoon; Oh, Sung; Oh, Tae-Kyun; Park, Jae Sung; Kim, Sei Chang; Kim, Seong Hwan; Kim, Young Shik; Hong, Jeum Kyu; Sim, Sang-Yun; Park, Kwon Seo; Lee, Hwan Gu; Kim, Kyung Jae; Choi, Chang Won
Background The highly pathogenic avian influenza H5N1 virus can infect a variety of animals and continually poses a threat to animal and human health. While many genotypes of H5N1 virus can be found in chicken, few are associated with the infection of mammals. Characterization of the genotypes of viral strains in animal populations is important to understand the distribution of different viral strains in various hosts. This also facilitates the surveillance and detection of possible emergence of highly pathogenic strains of specific genotypes from unknown hosts or hosts that have not been previously reported to carry these genotypes. Methodology/Principal Findings Two H5N1 isolates were obtained from lung samples of two raccoon dogs that had died from respiratory disease in China. Pathogenicity experiments showed that the isolates were highly pathogenic to chicken. To characterize the genotypes of these viruses, their genomic sequences were determined and analyzed. The genetic contents of these isolates are virtually identical and they may come from the same progenitor virus. Phylogenetic analysis indicated that the isolates were genetically closely related to genotype V H5N1 virus, which was first isolated in China in 2003, and were distinct from the dominant virus genotypes (e.g. genotype Z) of recent years. The isolates also contain a multibasic amino acid motif at their HA cleavage sites and have an E residue at position 627 of the PB2 protein similar to the previously-identified avian viruses. Conclusions/Significance This is the first report that genotype V H5N1 virus is found to be associated with a mammalian host. Our results strongly suggest that genotype V H5N1 virus has the ability to cross species barriers to infect mammalian animals. These findings further highlight the risk that avian influenza H5N1 virus poses to mammals and humans, which may be infected by specific genotypes that are not known to infect these hosts.
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. Virusisolates 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
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
Summary Genetic variation and phylogenetic relationships among 22 French isolates of equine arteritis virus (EAV) obtained over four\\u000a breeding seasons (2001–2004) were determined by sequencing open reading frames (ORFs) 2a–7. The ORFs 2a–7 of 22 isolates differed\\u000a from the prototype virulent Bucyrus strain of EAV by between 14 (99.5% identity) and 328 (88.7% identity) nucleotides, and\\u000a differed from each other by
J. Zhang; F. Miszczak; S. Pronost; C. Fortier; U. B. R. Balasuriya; S. Zientara; G. Fortier; P. J. Timoney
The spread of highly pathogenic avian influenza H5N1 viruses across Asia in 2003 and 2004 devastated domestic poultry populations and resulted in the largest and most lethal H5N1 virus outbreak in humans to date. To better understand the potential of H5N1 virusesisolated during this epizootic event to cause disease in mammals, we used the mouse and ferret models to
Taronna R. Maines; Xui Hua Lu; Steven M. Erb; Lindsay Edwards; Jeannette Guarner; Patricia W. Greer; Doan C. Nguyen; Kristy J. Szretter; Li-Mei Chen; Pranee Thawatsupha; Malinee Chittaganpitch; Sunthareeya Waicharoen; Diep T. Nguyen; Tung Nguyen; Hanh H. T. Nguyen; Jae-Hong Kim; Long T. Hoang; Chun Kang; Lien S. Phuong; Wilina Lim; Sherif Zaki; Ruben O. Donis; Nancy J. Cox; Jacqueline M. Katz; Terrence M. Tumpey
It is generally believed that pigs can serve as an intermediate host for the transmission of avian influenza viruses to humans or as mixing vessels for the generation of avian–human reassortant viruses. Here we describe the antigenic and genetic characterization of two influenza A (H1N1) viruses, which were isolated in The Netherlands from two patients who suffered from pneumonia. Both
G. F. Rimmelzwaan; J. C. de Jong; T. M. Bestebroer; A. M. van Loon; E. C. J. Claas; R. A. M. Fouchier; A. D. M. E. Osterhaus
In October 1999, H4N6 influenza A viruses were isolated from pigs with pneumonia on a commercial swine farm in Canada. Phylogenetic analyses of the sequences of all eight viral RNA segments demonstrated that these are wholly avian influenza viruses of the North American lineage. To our knowledge, this is the first report of interspecies transmission of an avian H4 influenza virus to domestic pigs under natural conditions.
Karasin, Alexander I.; Brown, Ian H.; Carman, Suzanne; Olsen, Christopher W.
Peste des Petits Ruminants (PPR) is an important viral disease of small ruminants and is endemic in Pakistan. In the following study, samples from two outbreaks of PPR in goats have been subjected to laboratory investigations. The Peste des Petits Ruminants virus (PPRV) genome was detected using both conventional and real-time PCR. Genetic characterization of the local PPRV field isolates was conducted by sequencing 322?bp of the fusion (F) gene and 255 bp of the nucleoprotein (N) gene. The phylogenetic tree based on the F gene clustered samples from both outbreaks into lineage 4 along with other Asian isolates, specifically into subcluster 1 along with isolates from Middle East. Analysis of N gene revealed a different pattern. In this case, the Pakistani samples clustered with Chinese, Tajikistani and Iranian isolates, which probably represents the true geographical pattern of virus circulation. This is the first report presenting the phylogenetic tree based on N gene as well as performing a parallel comparison of the trees of F and N gene together from Pakistani isolates. The results of this study shed light on the PPRV population in Pakistan and emphasize the importance of using molecular methods to understand the epidemiology. Such understanding is essential in any efforts to control the number and impact of outbreaks that are occurring in endemic countries such as Pakistan, especially in the current scenario where OIE and FAO are eager to control and subsequently eradicate PPR from the globe, as has been achieved for Rinderpest. PMID:21777402
Munir, M; Zohari, S; Saeed, A; Khan, Q M; Abubakar, M; LeBlanc, N; Berg, M
Summary The complete nucleotide sequence of a Chrysanthemum virus B isolate from Japan (CVB-S) has been determined. The genomic RNA of CVB-S is 8,990 nucleotides long, excluding the poly(A)\\u000a tail and, like that of other carlaviruses, contains six open reading frames (ORFs). Multiple alignment and phylogenetic analyses\\u000a indicated that the phylogenetic relationship among members of the genus Carlavirus is very diverse,
A. Ohkawa; M. Yamada; H. Sayama; N. Sugiyama; S. Okuda; T. Natsuaki
A novel human B-lymphotropic virus (HBLV) was isolated from the peripheral blood leukocytes of six individuals: two HTLV-III seropositive patients from the United States (one with AIDS-related lymphoma and one with dermatopathic lymphadenopathy), three HTLV-III seronegative patients from the United States (one with angioimmunoblastic lymphadenopathy, one with cutaneous T-cell lymphoma, and one with immunoblastic lymphoma), and one HTLV-III seronegative patient
S. Zaki Salahuddin; Dharam V. Ablashi; Phillip D. Markham; Steven F. Josephs; Susi Sturzenegger; Mark Kaplan; Gregory Halligan; Peter Biberfeld; Flossie Wong-Staal; Bernhard Kramarsky; Robert C. Gallo
Molecular characterization of Bovine leukemia virus (BLV) isolates from Brazil using the env gene sequences revealed a high conservation of this gene. In most cases the substitutions corresponded to silent transitions.\\u000a In addition, cystein residues, potential glycosylation sites, neutralization domains and other critical residues involved\\u000a with the envelope structural domains and viral infectivity were conserved. Most of the substitutions found
Marcelo Fernandes Camargos; Ariel Pereda; Daniel Stancek; Maurílio Andrade Rocha; Jenner Karlisson Pimenta dos Reis; Irene Greiser-Wilke; Rômulo Cerqueira Leite
BACKGROUND: The objective of this study is to investigate the pathogenesis of swine influenza virus (SIV) subtype H1N1 and H3N2 (Thai isolates) in 22-day-old SPF pigs. RESULTS: The study found that all pigs in the infected groups developed typical signs of flu-like symptoms on 1–4 days post- infection (dpi). The H1N1-infected pigs had greater lung lesion scores than those of
Human T-lymphotropic virus type I (HTLV-I) is associated with adult T-cell leukemia\\/lymphoma and with a chronic degenerative myelopathy. However, another major type of HTLV, HTLV-II, has been isolated only sporadically, and little is known of disease associations, transmission routes, and risk factors for HTLV-II infection. Recent studies indicate that a high percentage of certain groups of i.v. drug users and
Michael D. Lairmore; Steven Jacobson; Fernando Gracia; Barun K. de; Luis Castillo; Mario Larreategui; Beverly D. Roberts; Paul H. Levine; William A. Blattner; Johnathan E. Kaplan
Summary. Australian infectious bronchitis viruses (IBV) have undergone a separate evolution due to geographic isolation. Consequently,\\u000a changes occurring in Australian IBV illustrate, independently from other countries, types of variability that could occur\\u000a in emerging IBV strains. Previously, we have identified two distinct genetic groups of IBV, designated subgroups 1 and 2.\\u000a IBV strains of subgroup 1 have S1 and N proteins
Several isolates of a human type-C retrovirus belonging to one group, known as human T-cell leukemia virus (HTLV), have previously been obtained from patients with adult T-cell leukemia or lymphoma. The T-cell tropism of HTLV and its prevalence in the Caribbean basin prompted a search for it in patients with the epidemic T-cell immune deficiency disorder known as AIDS. Peripheral
Robert C. Gallo; Prem S. Sarin; E. P. Gelmann; Marjorie Robert-Guroff; Ersell Richardson; V. S. Kalyanaraman; Dean Mann; Gurdip D. Sidhu; Rosalyn E. Stahl; Susan Zolla-Pazner; Jacque Leibowitch; Mikulas Popovic
Rabies virus (RABV) strains Rus(Lipetsk)-8052f, Rus(Lipetsk)-8053c, Rus(Lipetsk)-8054f, and Rus(Lipetsk)-8057f were isolated from foxes (Vulpes vulpes) and a cat (Felis catus) in the Lipetsk region of Russia in 2011. Close relationships between these strains and the members of the "Cosmopolitan" group from Russia (98% homology) and from Europe (95% homology) were estimated. PMID:23661472
Poleshchuk, E M; Deviatkin, A A; Dedkov, V G; Sidorov, G N; Ochkasova, J V; Hodjakova, I A; Schukina, I A; Savel'ev, S I; Golenskih, A G; Shipulin, G A
Rabies virus (RABV) strains Rus(Lipetsk)-8052f, Rus(Lipetsk)-8053c, Rus(Lipetsk)-8054f, and Rus(Lipetsk)-8057f were isolated from foxes (Vulpes vulpes) and a cat (Felis catus) in the Lipetsk region of Russia in 2011. Close relationships between these strains and the members of the “Cosmopolitan” group from Russia (98% homology) and from Europe (95% homology) were estimated.
Poleshchuk, E. M.; Dedkov, V. G.; Sidorov, G. N.; Ochkasova, J. V.; Hodjakova, I. A.; Schukina, I. A.; Savel'ev, S. I.; Golenskih, A. G.; Shipulin, G. A.
Human immunodeficiency virus (HIV) was readily isolated by co-cultivation of patients' cells with phytohaemagglutinin-stimulated mononuclear cells from umbilical cord blood in 2 ml cultures in 24-well plates. Fluids from cultures of the MLA 144 cell line acted as an excellent source of interleukin-2, and promoted early replication of HIV in the primary cultures. Reverse transcriptase activity was commonly present at
JH Pope; F Bisshop; B McRandle; J Blok; CW Schmidt; RJ Kemp
Epidemic outbreaks of Tomato yellow leaf curl virus (TYLCV) diseases occurred in greenhouse grown tomato (Solanum lycopersicum) plants of Busan (TYLCV-Bus), Boseong (TYLCV-Bos), Hwaseong (TYLCV-Hwas), Jeju Island (TYLCV-Jeju), and Nonsan (TYLCV-Nons)\\u000a in Korea during 2008–2009. Tomato disease by TYLCV has never occurred in Korea before. We synthesized the full-length genomes\\u000a of each TYLCV isolate from the tomato plants collected at
Sue Hoon Kim; Sung Oh; Tae-Kyun Oh; Jae Sung Park; Sei Chang Kim; Seong Hwan Kim; Young Shik Kim; Jeum Kyu Hong; Sang-Yun Sim; Kwon Seo Park; Hwan Gu Lee; Kyung Jae Kim; Chang Won Choi
Members of the genus Yatapoxvirus, which include Tanapox virus (TPV) and Yaba monkey tumor virus, infect primates including humans. Two strains of TPV isolated 50 years apart from patients infected from the equatorial region of Africa have been sequenced. The original isolate from a human case in the Tana River Valley, Kenya, in 1957 (TPV-Kenya) and an isolate from an infected traveler in the Republic of Congo in 2004 (TPV-RoC). Although isolated 50 years apart the genomes were highly conserved. The genomes differed at only 35 of 144,565 nucleotide positions (99.98% identical). We predict that TPV-RoC encodes 155 ORFs, however a single transversion (at nucleotide 10241) in TPV-Kenya resulted in the coding capacity for two predicted ORFs (11.1L and 11.2L) in comparison to a single ORF (11L) in TPV-RoC. The genomes of TPV are A+T rich (73%) and 96% of the sequence encodes predicted ORFs. Comparative genomic analysis identified several features shared with other chordopoxviruses. A conserved sequence within the terminal inverted repeat region that is also present in the other members of the Yatapoxviruses as well as members of the Capripoxviruses, Swinepox virus and an unclassified Deerpox virus suggests the existence of a conserved near-terminal sequence secondary structure. Two previously unidentified gene families were annotated that are represented by ORF TPV28L, which matched homologues in certain other chordopoxviruses, and TPV42.5L, which is highly conserved among currently reported chordopoxvirus sequences. PMID:17574698
Nazarian, Steven H; Barrett, John W; Frace, A Michael; Olsen-Rasmussen, Melissa; Khristova, Marina; Shaban, Mae; Neering, Sarah; Li, Yu; Damon, Inger K; Esposito, Joseph J; Essani, Karim; McFadden, Grant
A Vero E6 cell culture isolate of Tula virus (TUL), a hantavirus first detected in European common voles (hficrotus arvalis and hr. rossiaemeridionolis) by RT-PCR was obtained after initial passaging of TUL- infected vole lung samples in laboratory-colonized hl. arvalis. TUL was defined as a classical serotype by a cross-focus-reduction neutralization test (FRNT) and was also shown to be distinct
Olli Vapalahti; A. Lundkvist; S. K. J. Kukkonen; Y. Cheng; M. Gilljam; M. Kanerva; T. Manni; M. Pejcoch; J. Niemimaa; A. Kaikusalo; H. Henttonen; A. Vaheri; A. Plyusnin
The population structure and genetic diversity of Cucumber vein yellowing virus (CVYV) from Spain were estimated by analyses of partial nucleotide sequences of the P1-proteinase (P1-Pro), P3 protein (P3),\\u000a and the coat protein (CP) coding regions. Analysis of 56 CVYV Spanish field isolates collected from 2001 to 2005 showed low\\u000a genetic diversity (0.0026, 0.0013, and 0.0012 for the P1-Pro, P3,
Dirk Janssen; Leonardo Velasco; Germán Martín; Eduardo Segundo; Isabel Maria Cuadrado
The complete nucleotide sequence of the blackgram isolate of mungbean yellow mosaic virus, IMYMV-Bg, which infects legumes in India, was determined and compared at the amino acid level with those of other whitefly-transmitted geminiviruses. The genome organization of IMYMV-Bg was similar to that of the begomoviruses. A unique feature of the genome organization was the sequence divergence of the common
V. Pant; D. Gupta; N. Roy Choudhury; V. G. Malathi; A. Varma; S. K. Mukherjee
SUMMARY Six cell fusion-causing syn mutants were isolated from the KOS (syn-lO1 to syn-106) and three from the HFEM (syn-107 to syn-109) strains of herpes simplex virus type 1 (HSV-1). The mutants were studied by complementation and recombination with syn-20 (a syncytial mutant of KOS) and ts-B5 (a syncytial mutant of HFEM). Some studies also employed MP, a syncytium-inducing strain
Porcine epidemic diarrhea virus (PEDV) has caused enteric disease with devastating impact since the first identification of\\u000a PEDV in 1992 in Korea. In this study, we investigated molecular epidemiology, showed genetic diversity, and analyzed phylogenetic\\u000a relationships of Korean PEDV field isolates with other PEDV reference strains. Genetic analysis of the complete M and ORF3\\u000a genes showed that each PEDV group
Seong-Jun Park; Hye-Kwon Kim; Dae-Sub Song; Hyoung-Joon Moon; Bong-Kyun Park
Porcine epidemic diarrhea virus (PEDV) causes a devastating enteric disease with acute diarrhea, dehydration and significant\\u000a mortality in swine, thereby incurring heavy economic losses in Korea. Spike (S) glycoprotein has been suggested as an important\\u000a determinant for PEDV biological properties. In this study, the nucleotide and deduced amino acid sequences of the partial\\u000a S glycoprotein genes of Korean PEDV isolates,
SUMMARY After several serial passages at low multiplicities of infection in primary human foetal glial cells at 37 °C, the DNA of prototype (MAD-I) JC virus and that of MAD-2 and MAD-3 are typically heterogeneous in size, but DNAs of MAD-4 and MAD-6 are relatively homogeneous. A similar dichotomy was observed in the DNAs of six isolates propagated more recently
JONATHAN D. MARTIN; BILLIE L. PADGETT; DUARD L. WALKER
The Cydia pomonella granulovirus (CpGV) has been used for many years as biological agent for codling moth control in apple orchards. Resistance to the Mexican\\u000a strain of CpGV was detected in orchards in Germany, France and Italy. A laboratory insect colony was started from insects\\u000a collected in a French resistant orchard. It was named RGV. Various virusisolates were identified
M. Berling; J.-B. Rey; S.-J. Ondet; Y. Tallot; O. Soubabčre; A. Bonhomme; B. Sauphanor; M. Lopez-Ferber
An influenza A virus (A/Tig/SH/01/2005 (H5N1) was isolated from lung tissue samples of a dead zoo tiger with respiratory disease in China in July 2005. Complete genome analysis indicated that the isolate was highly identical to an H5N1 virusisolated from a migratory duck at Poyang lake in China in that year. The genotype of the isolate was K,G,D,5J,F,1J,F,1E, and phylogenetically it was a clade 2.2 virus. Molecular characterization of all of the gene segments revealed characteristics of highly pathogenic influenza A viruses. These results may help to identify molecular determinants of virulence and highlight the necessity for continuous surveillance. PMID:18592132
Mushtaq, Muhammad Hassan; Juan, Huang; Jiang, Ping; Li, Yufeng; Li, TianXian; Du, Yijun; Mukhtar, Muhammad Mahmood
A study was performed in 2007 to isolate and characterize infectious bursal disease viruses (IBDVs) in commercial broilers grown in the Delmarva (DMV) Peninsula region of the United States. Bursae of Fabricius were collected weekly from 1 to 4 wk of age from broilers on 10 farms with a history of poor performance. Microscopic pathology was used to determine the infectious bursal disease (IBD) status of the broilers. Bursae from 1- and 2-wk-old broilers did not show IBD microscopic lesions. Moreover, broilers on 1 of the 10 farms were IBD lesion free at 3 and 4 wk of age. However, 3 of 9 and 9 of 9 farms yielded broilers with IBD-affected bursae from 3- and 4-wk-old commercial broilers, respectively. Ten IBDV isolates were recovered from 3 of 3 lesion-positive bursal pools at 3 wk of age and 7 of 9 lesion-positive bursal pools at 4 wk of age. Analysis of the viral protein (VP) 2 genes identified all isolates as serotype 1 Delaware (Del) variant viruses. Five field isolates, each representing different molecular clades of the Delaware variant viruses, were selected for further study. Experimental infection of specific-pathogen-free white leghorn chickens with isolates DMV/4813/07, DMV/4947/07, DMV/4955/07, DMV/5038/07, and DMV/5041/07 produced gross and microscopic pathology of the bursa consistent with Delaware variant infection. Monoclonal antibody testing showed DMV/4813/07, DMV/4947/07, DMV/ 4955/07, and DMV/5041/07 to be similar to previous recognized variant viruses. However, DMV/5038/07 was found to be unreactive with the monoclonal antibodies that typically recognize reference strains STC, Del E, GLS, RS593, and AL2. In a challenge of immunity study, 10-day-old progeny from breeders immunized with a commercially available inactivated IBDV vaccine containing the Del E and classic strains were protected to a lesser degree against isolate DMV/5038/07 compared to Del E challenge based on microscopic lesion scores (P < 0.01) of the bursa. This result suggests the virus is antigenically different from the Del E strain contained in the vaccine. Collectively, the monoclonal antibody and progeny challenge of immunity findings suggest DMV/5038/07 is antigenically different from the Del E strain contained in the vaccine. PMID:22545532
Gelb, J; Jackwood, D J; Mundt, E; Pope, C R; Hein, R; Slacum, G; Harris, J M; Ladman, B S; Lynch, P; Bautista, D A; Ruano, J M; Troeber, M M
The open reading frame 2 (ORF2) of three laboratory strains, the live attenuated vaccine virus, and 18 field isolates of equine arteritis virus (EAV) from Europe and North America was sequenced. The ORF2 of EAV encodes the Gs protein that is abundantly expressed in infected cells but constitutes less than 2% of the virion protein mass. Variation of ORF2 among
Jodi F. Hedges; Udeni B. R. Balasuriya; Peter J. Timoney; William H. McCollum; N. James MacLachlan
In this study, the full-genome sequence of a novel reassortant H1N1 swine influenza virus (SIV) is reported. The isolate has a hemagglutinin (HA) gene of the pandemic H1N1 influenza virus, but it carries the seven genome segments of the avian-origin H1N1 SIV currently circulating in European pig farms.
Chiapponi, Chiara; Baioni, Laura; Luppi, Andrea; Moreno, Ana; Castellan, Alberto
The pathogenicity of an extraneous subgroup A avian leukosis virus (ALV-A), isolated from commercial Marek's disease vaccines (Fadly et. al., Proc. USAHA; 2003; p.524-525) and termed B-39, was compared with that of Rous-associated virus-1 (RAV-1), the prototype strain of ALV-A. Chickens from ADOL li...
This study determined whether HLA-DR was incorporated into human immunodeficiency virus type 1 produced in vivo or by primary cultured cells. HLA-DR was associated with virions from primary isolates, macrophage cultures, and blood plasma. These results represent the first demonstration of major histocompatibility complex molecules associated with an in vivo source of virus.
Saarloos, M N; Sullivan, B L; Czerniewski, M A; Parameswar, K D; Spear, G T
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 virusisolation in embryonating chicken eggs (ECE) are standard methods for detecting A...
In this study, the full-genome sequence of a novel reassortant H1N1 swine influenza virus (SIV) is reported. The isolate has a hemagglutinin (HA) gene of the pandemic H1N1 influenza virus, but it carries the seven genome segments of the avian-origin H1N1 SIV currently circulating in European pig farms. PMID:24092781
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 virusisolates were recovered from birds of the order Columbiformes. ...
The cassava vein mosaic virus (CVMV) is a double stranded DNA virus which infects cassava plants (Manihot esculenta Crantz) and has been characterized as a plant pararetrovirus belonging to the caulimovirus subgroup. Two DNA fragments, CVP1 of 388 nucleotides from position -368 to +20 and CVP2 of 511 nucleotides from position -443 to +72, were isolated from the viral genome
Bertrand Verdaguer; Alexandre de Kochko; Roger N. Beachy; Claude Fauquet
European brown hare syndrome (EBHS) is characterised by high mortality of European brown hares (Lepus europaeus) and mountain hares (Lepus timidus). European brown hare syndrome virus (EBHSV) and the closely related rabbit haemorrhagic disease virus (RHDV) comprise the genus Lagovirus, family Caliciviridae. In contrast to RHDV, which is well studied, with more than 30 complete genome sequences available, the only complete genome sequence available for EBHSV was obtained from a strain isolated in 1989 in France. EBHS was originally diagnosed in Sweden in 1980. Here, we report the complete coding sequences of two EBHSV strains isolated from European brown hares that died with liver lesions characteristic of EBHS in Sweden in 1982. These sequences represent the oldest complete coding sequences of EBHSV isolated from the original area of virus diagnosis. The genomic organisation is similar to that of the published French sequence. Comparison with this sequence revealed several nucleotide substitutions, corresponding to 6 % divergence. At the amino acid level, the Swedish strains are 2 % different from the French strain. Most amino acid substitutions were located within the major capsid protein VP60, but when considering the amino acid sequence length of each protein, VP10 is the protein with the highest percentage of amino acid differences. The same result was obtained when Swedish strains were compared. This evolutionary pattern has not been described previously for members of the genus Lagovirus. PMID:23640583
Lopes, Ana M; Gavier-Widén, Dolores; Le Gall-Reculé, Ghislaine; Esteves, Pedro J; Abrantes, Joana
A continuous cell line, the flounder embryonic cell line (FEC), was established from gastrula-stage embryos of a marine cultured fish, the Japanese flounder Paralichthys olivaceus and cultured for more than 200 d with more than 60 passages. FEC cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with antibiotics, fetal bovine serum (FBS), sea perch serum (SPS), and basic fibroblast growth factor (bFGF). The cells were small and round, and grew actively and stably in culture. The effect of temperature, FBS concentration and bFGF on FEC cell growth was examined. Cells grew well between 24 and 30 degrees C, but had a reduced growth rate below 18 degrees C. The growth rate of FEC cells in medium containing 15% FBS was higher than that in medium containing 7.5% FBS. Addition of bFGF to the medium also significantly increased the growth rate. Chromosome analysis revealed that FEC cells have a normal diploid karyotype with 2n = 48. High survival rate was obtained after cryopreservation of cell cultures. The susceptibility of the cell line to piscine viruses was examined. Two viruses tested were shown to induce CPE (cytopathic effect) on FEC cells. FEC cell culture infected with fish iridovirus was further elucidated by electron microscopy. Many virus particles were found in the cytoplasm of the virus-infected FEC cells. These results indicated that the FEC cell line could be potentially used to isolate and study fish viruses. PMID:15521323
Virus replication and pulmonary disease pathogenesis in ferrets following intranasal infection with a pandemic influenza virus strain (A/California/4/09 [CA09]), a human seasonal influenza H1N1 virusisolate (A/New Caledonia/20/99 [Ncal99]), a classical swine influenza H1N1 virusisolate (A/Swine/Iowa/15/30 [Sw30]), or an avian H1N1 virusisolate (A/Mallard/MN/A108-2355/08 [Mal08]) were compared. Nasal wash virus titers were similar for Ncal99 and Sw30, with peak virus titers of 105.1 50% tissue culture infectious doses (TCID50)/ml and 105.5 TCID50/ml occurring at day 3 postinfection (p.i.), respectively. The mean peak titer for CA09 also occurred at day 3 p.i. but was higher (107 TCID50/ml). In contrast, the peak virus titers (103.6 to 104.3 TCID50/ml) for Mal08 were delayed, occurring between days 5 and 7 p.i. Disease pathogenesis was characterized by microscopic lesions in the nasal turbinates and lungs of all ferrets; however, Sw30 infection was associated with severe bronchointerstitial pneumonia. The results demonstrate that although CA09 is highly transmissible in the human population and replicates well in the ferret model, it causes modest disease compared to other H1N1 viruses, particularly Sw30 infection.
Smith, Jennifer Humberd; Nagy, Tamas; Driskell, Elizabeth; Brooks, Paula; Tompkins, S. Mark; Tripp, Ralph A.
SUMMARY. Avian influenza H5N1 viruses pose a significant threat to human health because of their ability to infect humans directly. In the paper, three highly pathogenic H5N1 influenza viruses were isolated from three species of migratory birds in Qinghai Province of China in 2006. The analysis of the genome sequences indicated that the three isolates shared high homology with each
Fumin Lei; AF Shuang Tang; A Xiaowei Zhang; Zhong Zhang; A Shengliang Chen; D Dehai Zhang; D Baoping Yan; Zuohua Yin; Shengliang Chen; Sandan Li; Dehai Zhang; Baoping Yan; Tianxian Li
BACKGROUND: The analysis of the nonstructural (NS) gene of the highly pathogenic (HP) H5N1 avian influenza viruses (AIV) isolated in Sweden early 2006 indicated the co-circulation of two sub-lineages of these viruses at that time. In order to complete the information on their genetic features and relation to other HP H5N1 AIVs the seven additional genes of twelve Swedish isolates
István Kiss; Péter Gyarmati; Siamak Zohari; Karin Wilbe Ramsay; Giorgi Metreveli; Elisabeth Weiss; Maria Brytting; Marielle Stivers; Sofia Lindström; Ake Lundkvist; Kirill Nemirov; Peter Thorén; Mikael Berg; György Czifra; Sándor Belák
The paper gives more detailed characteristics of Arabis mosaic virus (AMV) and tomato bushy stunt virus (TBSV) isolated from\\u000a lilac, the latter being identified in lilac (from plants suffering from yellow ring disease) for the first time. The isolate\\u000a of TBSV from lilac, from which an antiserum with a titre of 1024 was prepared, is closely related to the artichoke
Newcastle disease (ND) virus (APMV?1) isolates submitted to the International Reference Laboratory for ND were characterised antigenically by their ability to cause binding of mouse monoclonal antibodies (mAbs) to cell cultures infected with the isolate. Since the availability of the mAbs 1526 viruses have been examined using a panel of nine mAbs and 818 with an extended panel of 26
D. J. Alexander; R. J. Manvell; J. P. Lowings; K. M. Frost; M. S. Collins; P. H. Russell; J. E. Smith
Genes of an influenza A (H5N1) virus from a human in Hong Kong isolated in May 1997 were sequenced and found to be all avian-like (K. Subbarao et al., Science 279:393–395, 1998). Gene sequences of this human isolate were compared to those of a highly pathogenic chicken H5N1 influenza virusisolated from Hong Kong in April 1997. Sequence comparisons of all eight RNA segments from the two viruses show greater than 99% sequence identity between them. However, neither isolate’s gene sequence was closely (>95% sequence identity) related to any other gene sequences found in the GenBank database. Phylogenetic analysis demonstrated that the nucleotide sequences of at least four of the eight RNA segments clustered with Eurasian origin avian influenza viruses. The hemagglutinin gene phylogenetic analysis also included the sequences from an additional three human and two chicken H5N1 virusisolates from Hong Kong, and the isolates separated into two closely related groups. However, no single amino acid change separated the chicken origin and human origin isolates, but they all contained multiple basic amino acids at the hemagglutinin cleavage site, which is associated with a highly pathogenic phenotype in poultry. In experimental intravenous inoculation studies with chickens, all seven viruses were highly pathogenic, killing most birds within 24 h. All infected chickens had virtually identical pathologic lesions, including moderate to severe diffuse edema and interstitial pneumonitis. Viral nucleoprotein was most frequently demonstrated in vascular endothelium, macrophages, heterophils, and cardiac myocytes. Asphyxiation from pulmonary edema and generalized cardiovascular collapse were the most likely pathogenic mechanisms responsible for illness and death. In summary, a small number of changes in hemagglutinin gene sequences defined two closely related subgroups, with both subgroups having human and chicken members, among the seven viruses examined from Hong Kong, and all seven viruses were highly pathogenic in chickens and caused similar lesions in experimental inoculations.
Suarez, David L.; Perdue, Michael L.; Cox, Nancy; Rowe, Thomas; Bender, Catherine; Huang, Jing; Swayne, David E.
Recently, isolates of porcine reproductive and respiratory syndrome virus (PRRSV) that possess nucleotide sequences similar to European isolates have been reported in United States herds. The origin, diversity and prevalence of European-like North American PRRSV isolates in the U.S. remain unknown. Nucleotide sequence analysis of the 12kb ORF1 of a North American isolate, SDPRRS 01-08 (01-08), showed 93.7% identity with
Ying Fang; Dal-Young Kim; Susan Ropp; Pam Steen; Jane Christopher-Hennings; Eric A. Nelson; Raymond R. R. Rowland
Rapid alternative methods are required to evaluate easily acyclovir (ACV) sensitivity of clinical herpes simplex virus (HSV) isolates. The objective of this study was to screen 54 ACV-sensitive and 41 ACV-resistant clinical HSV-1 isolates, well characterized by phenotypic and genotypic methods, for the phosphorylation activity of the viral thymidine kinase (TK) using a commercially available and modified non-radioactive DiviTum® test on the basis of an indirect enzyme linked immunosorbent assay. The ACV-sensitive HSV-1 isolates had high TK activity values between 31.5±6.4 DiviTum® Units per liter (DU/L) and 487.4±60.1 DU/L. The mean activity of all ACV-sensitive isolates was calculated as 212.3±15.7 DU/L. By contrast, the mean activity of all ACV-resistant HSV-1 isolates was significantly lower at 5.5±1.3 DU/L. Out of the 41 ACV-resistant HSV-1 isolates, 38 had no or very low phosphorylation activities of the viral TK between 0 DU/L and 9.3±3.2 DU/L. The remaining three ACV-resistant viral isolates had TK activities between 44.6±5.1 DU/L and 80.9±13.3D U/L. In conclusion, the modified DiviTum® test can be used to screen HSV-1 isolates for their sensitivity to ACV. Acyclovir-sensitive HSV-1 isolates show TK activities >30 DU/L and ACV-resistant isolates have activity values <10 DU/L. However, single ACV-resistant HSV-1 isolates can have TK activity values >30 DU/L. These strains are most likely ACV-resistant TK-altered mutants, but no evidence was provided for an alteration of the TK. PMID:23246510
Sauerbrei, Andreas; Vödisch, Susanne; Bohn, Kathrin; Schacke, Michael; Gronowitz, Simon
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, virusisolation was performed from the tissue homogenates of the heart and lung obtained from the two juvenile orang utans in Vero cell cultures. The tissue culture fluid was examined using electron microscopy. Reverse transcription and polymerase chain reaction with Encephalomyocarditis virus (EMCV)-specific primers targeting the gene regions of VP3/VP1 and 3D polymerase (3Dpol) confirmed the virus genus and species. The two EMCV isolates were sequenced and phylogenetic analyses of the virus genes performed. Serological testing on other animal species in the Singapore Zoological Gardens was also conducted. Results Electron microscopy of the two EMCV isolates, designated Sing-M100-02 and Sing-M105-02, revealed spherical viral particles of about 20 to 30 nm, consistent with the size and morphology of members belonging to the family Picornaviridae. In addition, infected-Vero cells showed positive immunoflorescence staining with antiserum to EMCV. Sequencing of the viral genome showed that the two EMCV isolates were 99.9% identical at the nucleotide level, indicating a similar source of origin. When compared with existing EMCV sequences in the VP1 and 3Dpol gene regions, the nucleotide divergence were at a maximum of 38.8% and 23.6% respectively, while the amino acid divergence were at a maximum of 33.9% and 11.3% respectively. Phylogenetic analyses of VP1 and 3Dpol genes further grouped the Sing-M100-02 and Sing-M105-02 isolates to themselves, away from existing EMCV lineages. This strongly suggested that Sing-M100-02 and Sing-M105-02 isolates are highly divergent variants of EMCV. Apart from the two deceased orang utans, a serological survey conducted among other zoo animals showed that a number of other animal species had neutralizing antibodies to Sing-M105-02 isolate, indicating that the EMCV variant has a relatively wide host range. Conclusions The etiological agent responsible for the fatal myocarditis cases among two of the four orang utans in the Singapore Zoological Gardens was a highly divergent variant of EMCV. This is the first report of an EMCV infection in Singapore and South East Asia.
H5N1 influenza A viruses are widely distributed among poultry in Asia, but until recently, only a limited number of wild birds were affected. During late April through June 2005, an outbreak of H5N1 virus infection occurred among wild birds at Qinghai Lake in China. Here, we describe the features of this outbreak. First identified in bar-headed geese, the disease soon spread to other avian species populating the lake. Sequence analysis of 15 viruses representing six avian species and collected at different times during the outbreak revealed four different H5N1 genotypes. Most of the isolates possessed lysine at position 627 in the PB2 protein, a residue known to be associated with virulence in mice and adaptation to humans. However, neither of the two index viruses possessed this residue. All of the viruses tested were pathogenic in mice, with the exception of one index virus. We also tested the replication of two virusesisolated during the Qinghai Lake outbreak and one unrelated duck H5N1 virus in rhesus macaques. The Qinghai Lake viruses did not replicate efficiently in these animals, producing no evidence of disease other than transient fever, while the duck virus replicated in multiple organs and caused symptoms of respiratory illness. Importantly, H5N1 virusesisolated in Mongolia, Russia, Inner Mongolia, and the Liaoning Province of China after August 2005 were genetically closely related to one of the genotypes isolated during the Qinghai outbreak, suggesting the dominant nature of this genotype and underscoring the need for worldwide intensive surveillance to minimize its devastating consequences. PMID:16731936
White tail disease (WTD) is found to cause immense economic losses in hatcheries and farms, with mortalities often reaching 100% within 2 or 3 days. The pathogenic agents have been identified as Macrobrachium rosenbergii nodavirus (MrNV) associated with extra small virus (XSV), which are 27 and 15 nm in diameter, respectively. Experiments were carried out to characterize an Indian isolate of XSV capsid protein of 17 kDa (CP-17). The gene encoding CP-17 was cloned and its sequence analysed with sequences of other isolates such as French, Chinese, Taiwanese and Thai available in the GenBank using Bioinformatics tools such as BLASTn, clustal W and phylogenetic analysis. When compared with other isolates, 18-point mutations were observed in Indian isolate (XSV-IN) with few changes in amino acid residues. Homology comparison showed 99-96% identity with other isolates. Phylogenetic analysis revealed that the Indian isolate was closely related to Taiwanese and Chinese isolates than French and Thai. This shows that the possible origin of the disease in India was from Taiwan and China through the import of prawn seed decades ago. PMID:17928085
Sudhakaran, R; Syed Musthaq, S; Rajesh Kumar, S; Sarathi, M; Sahul Hameed, A S