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multiplication of venezuelan equine encephalitis (mucambo) virus in cultured mosquito cells.venezuelan equine encephalitis (mucambo) virus was able to multiply at 28degree c in cultured mosquito cells (aedes aegypti and aedes albopictus) without inducing any apparent cytopathology. the kinetics of virus multiplication was similar in mosquito and mammalian (vero) cells. twelve to twenty four hours post-infection, a maximum of 10-50 per cent of the cells were engaged in virus production; however, almost immediately this proportion fell abruptly tending to stabilize in the order of 0.01 t ...1975813617
induction of venezuelan equine encephalitis (mucambo) virus by iododeoxyuridine in chronically infected 'cured' cultured mosquito cells.infection of cultured aedes aegypti mosquito cells with venezuelan equine encephalitis virus (mucambo subtype) results in the establishment of a chronic non-cytopathic infection. infected cells could be 'cured' when they were grown in the presence of immune serum for 3-4 passages. 'cured' cells could then be subcultured repeatedly, in the absence of immune serum, without showing any indication of infectious virus production. when 'cured' cells were exposed to 50 microgram/ml of 5-iodo-2-deoxyuri ...1977873741
evolutionary relationships and systematics of the alphaviruses.partial e1 envelope glycoprotein gene sequences and complete structural polyprotein sequences were used to compare divergence and construct phylogenetic trees for the genus alphavirus. tree topologies indicated that the mosquito-borne alphaviruses could have arisen in either the old or the new world, with at least two transoceanic introductions to account for their current distribution. the time frame for alphavirus diversification could not be estimated because maximum-likelihood analyses indic ...200111581380
evolutionary genetics and vector adaptation of recombinant viruses of the western equine encephalitis antigenic complex provides new insights into alphavirus diversity and host switching.western equine encephalitis virus (weev), highlands j virus (hjv), and fort morgan virus (fmv) are the sole representatives of the wee antigenic complex of the genus alphavirus, family togaviridae, that are endemic to north america. all three viruses have their ancestry in a recombination event involving eastern equine encephalitis virus (eeev) and a sindbis (sin)-like virus that gave rise to a chimeric alphavirus that subsequently diversified into the present-day weev, hjv, and fmv. here, we pr ...201425463613
in- silico exploration of thirty alphavirus genomes for analysis of the simple sequence repeats.the compilation of simple sequence repeats (ssrs) in viruses and its analysis with reference to incidence, distribution and variation would be instrumental in understanding the functional and evolutionary aspects of repeat sequences. present study encompasses the analysis of ssrs across 30 species of alphaviruses. the full length genome sequences, assessed from ncbi were used for extraction and analysis of repeat sequences using imex software. the repeats of different motif sizes (mono- to penta ...201425606453
isolation and characterisation of a human-like antibody fragment (scfv) that inactivates veev in vitro and in vivo.venezuelan equine encephalitis virus (veev) belongs to the alphavirus genus and several species of this family are pathogenic to humans. the viruses are classified as potential agents of biological warfare and terrorism and sensitive detection as well as effective prophylaxis and antiviral therapies are required.in this work, we describe the isolation of the anti-veev single chain fragment variable (scfv), tor67-3b4, from a non-human primate (nhp) antibody gene library. we report its recloning i ...201222666347
a humanised murine monoclonal antibody with broad serogroup specificity protects mice from challenge with venezuelan equine encephalitis virus.in murine models of venezuelan equine encephalitis virus (veev) infection, the neutralising monoclonal antibody 1a3b-7 has been shown to be effective in passive protection from challenge by the aerosol route with serogroups i, ii and mucambo virus (formally vee complex subtype iiia). this antibody is able to bind to all serogroups of the veev complex when used in elisa and therefore is an excellent candidate for protein engineering in order to derive a humanised molecule suitable for therapeutic ...201121310183
isolation and characterization of sylvatic mosquito-borne viruses in trinidad: enzootic transmission and a new potential vector of mucambo virus.mosquito surveillance was carried out in three forested regions of trinidad during july 2007-march 2009. a total of 185,397 mosquitoes representing at least 46 species was collected, divided into pools of 1-50 mosquitoes according to species and sex, and screened for arboviruses using cytopathic effect assays on vero cell monolayers. eighty-five viruses were isolated, including members of the genera alphavirus (mucambo virus; mucv) and orthobunyavirus (caraparu, oriboca, bimiti, and wyeomyia vir ...201021118932
comparison of the immunological responses and efficacy of gamma-irradiated v3526 vaccine formulations against subcutaneous and aerosol challenge with venezuelan equine encephalitis virus subtype iab.we recently developed a gamma-irradiation method to inactivate v3526, a live-attenuated venezuelan equine encephalitis virus (veev) vaccine candidate. dosage and schedule studies were conducted to evaluate the immunogenicity and efficacy of gamma-irradiated v3526 (gv3526). subcutaneous (sc) and low dosage intramuscular (im) administration of gv3526 were highly effective in protecting mice against a sc challenge with veev ia/b trinidad donkey strain, but not against an equivalent aerosol challeng ...201019914193
evolutionary patterns of eastern equine encephalitis virus in north versus south america suggest ecological differences and taxonomic revision.the eastern equine encephalitis (eee) complex consists of four distinct genetic lineages: one that circulates in north america (na eeev) and the caribbean and three that circulate in central and south america (sa eeev). differences in their geographic, pathogenic, and epidemiologic profiles prompted evaluation of their genetic diversity and evolutionary histories. the structural polyprotein open reading frames of all available sa eeev and recent na eeev isolates were sequenced and used in evolut ...201019889755
isolation and phylogenetic analysis of mucambo virus (venezuelan equine encephalitis complex subtype iiia) in trinidad.in the 1950s and 1960s, alphaviruses in the venezuelan equine encephalitis (vee) antigenic complex were the most frequently isolated arboviruses in trinidad. since then, there has been very little research performed with these viruses. herein, we report on the isolation, sequencing, and phylogenetic analyses of mucambo virus (mucv; vee complex subtype iiia), including 6 recently isolated from culex (melanoconion) portesi mosquitoes and 11 previously isolated in trinidad and brazil. results show ...200919631956
duplex reverse transcription-pcr followed by nested pcr assays for detection and identification of brazilian alphaviruses and flaviviruses.a new approach was developed for the rapid detection and identification of brazilian alphaviruses and flaviviruses. the methodology involves the genus-specific detection of alphavirus and flavivirus by a duplex reverse transcription-pcr (d-rt-pcr), followed by multiplex nested pcr (m-n-pcr) or nested pcr (n-pcr) assays for species-specific identification. by this protocol, 25 arboviruses were specifically detected and identified. detection levels between 10(1.3) and 10(3.5) 50% tissue culture in ...200515695666
reverse transcription-pcr-enzyme-linked immunosorbent assay for rapid detection and differentiation of alphavirus infections.due to the lack of a rapid, simple, and inexpensive assay for detecting alphavirus infections, we combined a reverse transcription-pcr with an enzyme-linked immunosorbent assay (rt-pcr-elisa) to identify human pathogenic alphaviruses that are endemic in the new world. by combining the sensitivity of pcr, the detection simplicity of elisa, and the specificities of dna probes, this method rapidly detected and differentiated closely related species and subtypes of several medically important alphav ...200616957044
directed molecular evolution improves the immunogenicity and protective efficacy of a venezuelan equine encephalitis virus dna vaccine.we employed directed molecular evolution to improve the cross-reactivity and immunogenicity of the venezuelan equine encephalitis virus (veev) envelope glycoproteins. the dna encoding the e1 and e2 proteins from veev subtypes ia/b and ie, mucambo virus (mucv), and eastern and western equine encephalitis viruses (eeev and weev) were recombined in vitro to create libraries of chimeric genes expressing variant envelope proteins. elisas specific for all five parent viruses were used in high-throughp ...200919406186
equine amplification and virulence of subtype ie venezuelan equine encephalitis viruses isolated during the 1993 and 1996 mexican epizootics.to assess the role of horses as amplification hosts during the 1993 and 1996 mexican venezuelan equine encephalitis (vee) epizootics, we subcutaneously infected 10 horses by using four different equine isolates. most horses showed little or no disease and low or nonexistent viremia. neurologic disease developed in only 1 horse, and brain histopathologic examination showed meningeal lymphocytic infiltration, perivascular cuffing, and focal encephalitis. three animals showed mild meningoencephalit ...200312603985
natural enzootic vectors of venezuelan equine encephalitis virus, magdalena valley, colombia.to characterize the transmission cycle of enzootic venezuelan equine encephalitis virus (veev) strains believed to represent an epizootic progenitor, we identified natural vectors in a sylvatic focus in the middle magdalena valley of colombia. hamster-baited traps were placed into an active forest focus, and mosquitoes collected from each trap in which a hamster became infected were sorted by species and assayed for virus. in 18 cases, a single, initial, high-titered mosquito pool representing t ...200312533281
endemic venezuelan equine encephalitis in northern peru.since venezuelan equine encephalitis virus (veev) was isolated in peru in 1942, >70 isolates have been obtained from mosquitoes, humans, and sylvatic mammals primarily in the amazon region. to investigate genetic relationships among the peru veev isolates and between the peru isolates and other veev strains, a fragment of the pe2 gene was amplified and analyzed by single-stranded conformation polymorphism. representatives of seven genotypes underwent sequencing and phylogenetic analysis. the res ...200415200823
two human cases of laboratory infection with mucambo virus. 196514292756
direct broad-range detection of alphaviruses in mosquito extracts.members of the genus alphavirus are a diverse group of principally mosquito-borne rna viruses. there are at least 29 species and many more subtypes of alphaviruses and some are considered potential bioweapons. we have developed a multi-locus rt-pcr followed by electrospray ionization mass spectrometry (rt-pcr/esi-ms) assay that uses the amplicon base compositions to detect and identify alphaviruses. a small set of primer pairs targeting conserved sites in the alphavirus rna genome were used to a ...200717655905
current strategic thinking for the development of a trivalent alphavirus vaccine for human use.vaccinations against the encephalitic alphaviruses (western, eastern, and venezuelan equine encephalitis virus) are of significant interest to biological defense, public health, and agricultural communities alike. although vaccines licensed for veterinary applications are used in the western hemisphere and attenuated or inactivated viruses have been used under investigational new drug status to protect at-risk personnel, there are currently no licensed vaccines for use in humans. here, we will d ...201424842880
serological evidence of flaviviruses and alphaviruses in livestock and wildlife in trinidad.seroprevalence rates of selected arboviruses in animal populations in trinidad were determined using serum samples collected between 2006 and 2009 from horses (n=506), cattle (n=163), sheep (n=198), goats (n=82), pigs (n=184), birds (n=140), rodents (n=116), and other vertebrates (n=23). the sera were screened for antibodies to west nile virus (wnv), st. louis encephalitis virus (slev), ilheus virus (ilhv), bussuquara virus (bsqv), venezuelan equine encephalitis virus (veev), eastern equine ence ...201222989182
diagnosis of arboviruses using indirect sandwich igg elisa in horses from the brazilian amazon.the amazon as a whole is the largest reservoir of arboviruses worldwide, while the brazilian amazon hosts the largest variety of arboviruses isolated to date. in this study, the results of an indirect sandwich igg elisa, standardized for 19 arbovirustypes circulating among horses in brazilian amazon, were compared to results of the hemagglutination inhibition test. a screening test assessed the conditional probability distribution and a pearson linear correlation test determined the correlation ...201425013410
hierarchical cluster analysis of three-dimensional reconstructions of unbiased sampled microglia shows not continuous morphological changes from stage 1 to 2 after multiple dengue infections in callithrix penicillata.it is known that microglial morphology and function are related, but few studies have explored the subtleties of microglial morphological changes in response to specific pathogens. in the present report we quantitated microglia morphological changes in a monkey model of dengue disease with virus cns invasion. to mimic multiple infections that usually occur in endemic areas, where higher dengue infection incidence and abundant mosquito vectors carrying different serotypes coexist, subjects receiv ...201627047345
sindbis and middelburg old world alphaviruses associated with neurologic disease in horses, south africa.old world alphaviruses were identified in 52 of 623 horses with febrile or neurologic disease in south africa. five of 8 sindbis virus infections were mild; 2 of 3 fatal cases involved co-infections. of 44 middelburg virus infections, 28 caused neurologic disease; 12 were fatal. middelburg virus likely has zoonotic potential.026583836
search for subtype i strains of venezuelan encephalitis virus among isolates from trinidad, w.i., during 1960-1972.twenty-one strains of venezuelan encephalitis (ve) virus isolated from three habitats in trinidad, w.i. during 1960 to 1972, were subtype iii (mucambo) ve virus by plaque-reduction neutralization tests. like prototype mucambo virus, each strain killed 8- to 15-week-old mice inoculated intraperitoneally. if the subtype i strain of ve virus that caused a major outbreak in trinidad during 1943 to 1944 persisted on the island into the 1960s and early 1970s, it did not become the dominant ve virus in ...1977842779
isolation of mucambo virus, a member of the venezuelan equine encephalitis virus complex in the state of são paulo, brasil. 1978675033
neutralising antibodies for mayaro virus in pantanal, brazil.the pantanal hosts diverse wildlife species and therefore is a hotspot for arbovirus studies in south america. a serosurvey for mayaro virus (mayv), eastern (eeev), western (weev) and venezuelan (veev) equine encephalitis viruses was conducted with 237 sheep, 87 free-ranging caimans and 748 equids, including 37 collected from a ranch where a neurologic disorder outbreak had been recently reported. sera were tested for specific viral antibodies using plaque-reduction neutralisation test. from a t ...201525742272
venezuelan equine encephalitis virus activity in the gulf coast region of mexico, 2003-2010.venezuelan equine encephalitis virus (veev) has been the causative agent for sporadic epidemics and equine epizootics throughout the americas since the 1930s. in 1969, an outbreak of venezuelan equine encephalitis (vee) spread rapidly from guatemala and through the gulf coast region of mexico, reaching texas in 1971. since this outbreak, there have been very few studies to determine the northward extent of endemic veev in this region. this study reports the findings of serologic surveillance in ...201223133685
[alterations of mouse kidney following inoculation of mucambo-virus]. 19715316272
transcriptome sequencing and annotation for the jamaican fruit bat (artibeus jamaicensis).the jamaican fruit bat (artibeus jamaicensis) is one of the most common bats in the tropical americas. it is thought to be a potential reservoir host of tacaribe virus, an arenavirus closely related to the south american hemorrhagic fever viruses. we performed transcriptome sequencing and annotation from lung, kidney and spleen tissues using 454 and illumina platforms to develop this species as an animal model. more than 100,000 contigs were assembled, with 25,000 genes that were functionally an ...201223166587
a dna vaccine for venezuelan equine encephalitis virus delivered by intramuscular electroporation elicits high levels of neutralizing antibodies in multiple animal models and provides protective immunity to mice and nonhuman primates.we evaluated the immunogenicity and protective efficacy of a dna vaccine expressing codon-optimized envelope glycoprotein genes of venezuelan equine encephalitis virus (veev) when delivered by intramuscular electroporation. mice vaccinated with the dna vaccine developed robust veev-neutralizing antibody responses that were comparable to those observed after administration of the live-attenuated veev vaccine tc-83 and were completely protected from a lethal aerosol veev challenge. the dna vaccine ...201121450977
antibody to the e3 glycoprotein protects mice against lethal venezuelan equine encephalitis virus infection.six monoclonal antibodies were isolated that exhibited specificity for a furin cleavage site deletion mutant (v3526) of venezuelan equine encephalitis virus (veev). these antibodies comprise a single competition group and bound the e3 glycoprotein of veev subtype i viruses but failed to bind the e3 glycoprotein of other alphaviruses. these antibodies neutralized v3526 virus infectivity but did not neutralize the parental strain of trinidad donkey (trd) veev. however, the e3-specific antibodies d ...201020926570
comprehensive biothreat cluster identification by pcr/electrospray-ionization mass spectrometry.technology for comprehensive identification of biothreats in environmental and clinical specimens is needed to protect citizens in the case of a biological attack. this is a challenge because there are dozens of bacterial and viral species that might be used in a biological attack and many have closely related near-neighbor organisms that are harmless. the biothreat agent, along with its near neighbors, can be thought of as a biothreat cluster or a biocluster for short. the ability to comprehens ...201222768032
a humanised murine monoclonal antibody protects mice from venezuelan equine encephalitis virus, everglades virus and mucambo virus when administered up to 48 h after airborne challenge.currently there are no licensed antiviral treatments for the alphaviruses venezuelan equine encephalitis virus (veev), everglades virus and mucambo virus. we previously developed a humanised version of the mouse monoclonal antibody 1a3b-7 (hu1a3b-7) which exhibited a wide range of reactivity in vitro and was able to protect mice from infection with veev. continued work with the humanised antibody has now demonstrated that it has the potential to be a new human therapeutic. hu1a3b-7 successfully ...201222341308
a multisystem approach for development and evaluation of inactivated vaccines for venezuelan equine encephalitis virus (veev).a multisystem approach was used to assess the efficiency of several methods for inactivation of venezuelan equine encephalitis virus (veev) vaccine candidates. a combination of diverse assays (plaque, in vitro cytopathology and mouse neurovirulence) was used to verify virus inactivation, along with the use of a specific elisa to measure retention of veev envelope glycoprotein epitopes in the development of several inactivated veev candidate vaccines derived from an attenuated strain of veev (v35 ...201019903494
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