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J Boshell

Publications and source records attributed to J Boshell.

At least 19 recordsLinked to original sources
PubMed2001

Genetic diversity and relationships among Venezuelan equine encephalitis virus field isolates from Colombia and Venezuela.

During field studies of enzootic Venezuelan equine encephalitis (VEE) viruses associated with epizootic emergence, a large number of virus isolates were made in sylvatic foci of Venezuela and Colombia. To rapidly characterize these isolates, antigenic subtypes were determined by means of immunofluorescence and by single-strand conformational polymorphism (SSCP) analysis by use of an 856-bp fragment from the P62 gene, which we used to distinguish genetic variants. Representative isolates were sequenced to assess the sensitivity of SSCP to detect genetic differences. The SSCP analysis distinguished isolates differing by as little as 1 nucleotide; overall, differences of > or = 1 nucleotide were recognized 89% of the time, and the sensitivity to distinguish strains that differed by only 1 or 4 nucleotides was 17 and 57%, respectively. Phylogenetic analyses of representative sequences showed that all recent isolates from the Catatumbo region of western Venezuela and the middle Magdalena Valley of Colombia were closely related to epizootic subtype IAB and IC strains; strains from Yaracuy and Miranda States were more distantly related. Cocirculation of the same virus genotype in both Colombian and Venezuelan foci indicated that these viruses are readily transported between enzootic regions separated by > 300 km. The SSCP analysis appears to be a simple, fast, and relatively efficient method of screening VEE virus isolates to identify meaningful genetic variants.

Aedes↗
PubMed1999

Genetic and phenotypic changes accompanying the emergence of epizootic subtype IC Venezuelan equine encephalitis viruses from an enzootic subtype ID progenitor.

Recent studies have indicated that epizootic Venezuelan equine encephalitis (VEE) viruses can evolve from enzootic, subtype ID strains that circulate continuously in lowland tropical forests (A. M. Powers, M. S. Oberste, A. C. Brault, R. Rico-Hesse, S. M. Schmura, J. F. Smith, W. Kang, W. P. Sweeney, and S. C. Weaver, J. Virol. 71:6697-6705, 1997). To identify mutations associated with the phenotypic changes leading to epizootics, we sequenced the entire genomes of two subtype IC epizootic VEE virus strains isolated during a 1992-1993 Venezuelan outbreak and four sympatric, subtype ID enzootic strains closely related to the predicted epizootic progenitor. Analysis by maximum-parsimony phylogenetic methods revealed 25 nucleotide differences which were predicted to have accompanied the 1992 epizootic emergence; 7 of these encoded amino acid changes in the nsP1, nsP3, capsid, and E2 envelope glycoprotein, and 2 were mutations in the 3' untranslated genome region. Comparisons with the genomic sequences of IAB and other IC epizootic VEE virus strains revealed that only one of the seven amino acid changes associated with the 1992 emergence, a threonine-to-methionine change at position 360 of the nsP3 protein, accompanied another VEE virus emergence event. Two changes in the E2 envelope glycoprotein region believed to include the major antigenic determinants, both involving replacement of uncharged residues with arginine, are also candidates for epizootic determinants.

Amino Acid Sequence↗
PubMed1997

Origins of dengue type 2 viruses associated with increased pathogenicity in the Americas.

The recent emergence and spread of dengue hemorrhagic fever in the Americas have been a major source of concern. Efforts to control this disease are dependent on understanding the pathogenicity of dengue viruses and their transmission dynamics. Pathogenicity studies have been hampered by the lack of in vitro or in vivo models of severe dengue disease. Alternatively, molecular epidemiologic studies which associate certain dengue virus genetic types with severe dengue outbreaks may point to strains with increased pathogenicity. The comparison of nucleotide sequences (240 bp) from the E/NS1 gene region of the dengue virus genome has been shown to reflect evolutionary relationships and geographic origins of dengue virus strains. This approach was used to demonstrate an association between the introduction of two distinct genotypes of dengue type 2 virus and the appearance of dengue hemorrhagic fever in the Americas. Phylogenetic analyses suggest that these genotypes originated in Southeast Asia and that they displaced the native, American genotype in at least four countries. Vaccination and other control efforts should therefore be directed at decreasing the transmission of these "virulent" genotypes.

Base Sequence↗
PubMed1997

Epidemic Venezuelan equine encephalitis in La Guajira, Colombia, 1995.

In 1995, the first Venezuelan equine encephalitis (VEE) outbreak in Colombia in 22 years caused an estimated 75,000 human cases, 3000 with neurologic complications and 300 fatal, in La Guajira State. Of the state's estimated 50,000 equines, 8% may have died. An epizootic IC virus, probably introduced from Venezuela, was rapidly amplified among unvaccinated equines. Record high rainfall, producing high densities of vector Aedes taeniorhynchus, led to extensive epidemic transmission (30% attack rate) in the four affected municipalities. Native Wayuu Indians, constituting 24% of the state's population, were at increased risk of infection (risk ratio, 3.3; 95% confidence interval, 2.2-5.3). Epidemiologic studies found no evidence of human-to-human transmission. A higher-than-expected number of abortions during the outbreak confirmed a previously suspected abortifacient role of VEE infection. Pesticide applications and a mass equine vaccination program contributed to preventing the outbreak's spread south of La Guajira.

Adult↗
PubMed1997

Immunoglobulin M antibody response to measles virus following natural virus infection, primary vaccination, and reexposure to the virus.

Evaluation of the measles virus ELISA kit (Merck) to detect specific IgM as an indicator of primary measles antibody response was carried out. A modification of the manufacturer's cutoff value interpretation was introduced to allow for equivocal results in addition to positive and negative ones. With this modification, the test assayed gave an overall reproducibility of 96.16%. The IgM seropositivity rate for seroneutralization-confirmed measles cases was 100% for naturally infected measles subjects and 90% for primary measles vaccinated subjects. Individuals with positive neutralizing antimeasles antibodies in close contact with a confirmed measles case gave the following measles IgM ELISA results: 54.54% negative, 9.09% positive, and 36.36% equivocal, showing a booster with IgM antibody response on reexposure to the virus. Positive subjects with neutralizing antimeasles antibodies without recent contact with a measles case gave negative IgM results. IgM seropositivity was strongly associated with IgG seroconversion and clinical measles (p < 0.0001). The technique assayed performed adequately for the confirmation of both measles natural infection and primary vaccination and for the differentiation of primary and secondary antibody response, taking into account the modification in the cutoff value interpretation introduced and providing that the serum samples are obtained between days 5 and 30 after onset of rash.

Adolescent↗
PubMed1997

Genotype-specific RNA probes for direct identification of wild polioviruses by blot hybridization.

We have developed RNA probes for the direct identification of wild poliovirus isolates by blot hybridization. The probes are complementary to sequences of the first 30 to 32 codons of VP1, which evolve more extensively (approximately 1.5-fold) than the rest of VP1. To illustrate our general approach, we describe the design of probes specific to each of four major genotypes recently endemic (1981 to 1991) to the Americas: Andean type 1, Brazil type 1, Brazil type 3, and Central America-Mexico type 3. A wild isolate of each genotype was selected according to molecular and epidemiologic criteria to be representative of the principal lineages in circulation. Variable VP1 sequences of the representative isolates were amplified by the reverse transcriptase PCR and were inserted into a plasmid vector containing a T7 promoter. The in vitro transcripts, labeled with digoxigenin, served as probes. These formed stable hybrids only with RNAs of isolates of the corresponding genotypes. Hybrids were detected by a sensitive chemiluminescence assay, capable under normal diagnostic conditions of detecting specific wild poliovirus sequences in samples containing up to a 100-fold excess of Sabin vaccine strain-related sequences of the same serotype.

Base Sequence↗
PubMed1996

Re-emergence of epidemic Venezuelan equine encephalomyelitis in South America. VEE Study Group.

BACKGROUND: Venezuelan equine encephalomyelitis (VEE) virus has caused periodic epidemics among human beings and equines in Latin America from the 1920s to the early 1970s. The first major outbreak since 1973 occurred in Venezuela and Colombia during 1995, and involved an estimated 75,000 to 100,000 people. We report an epidemiological and virological investigation of this epidemic. METHODS: Virus isolates were made in cell culture from human serum, human throat swabs, and brain tissue from aborted and stillborn human fetuses, as well as from horse brain tissue and pooled mosquito collections. Human sera were also tested for VEE-specific antibodies. The serotypes of VEE isolates were identified by antigen assays, and viruses were characterised genetically by sequencing PCR products generated from the E3 and E2 genes. Phylogenetic analyses were done to determine evolutionary relations with respect to previous epidemic/epizootic and enzootic VEE virus isolates. Mosquito collections were made to identify possible vectors, and clinical findings were determined by direct observation of patients visiting hospitals and clinics in affected regions, and by inspecting patient records. Equine vaccination and vector control were used in an attempt to halt the spread of the outbreak. FINDINGS: Most affected people had an acute, self-limited febrile illness of 3 to 4 days duration. However, convulsions were often seen in children, and abortions and fetal deaths occurred in pregnant women infected with VEE virus. Antigenic characterisation of 12 virus isolates spanning the temporal and spatial range of the outbreak indicated that all are VEE serotype IC. Phylogenetic analysis revealed that all of the 1995 viruses were closely related to serotype IC viruses isolated during a large VEE outbreak that occurred in the same regions of Colombia and Venezuela from 1962-1964. A 1983 mosquito isolate from north central Venezuela was also closely related to the 1995 isolates. INTERPRETATION: This outbreak was remarkably similar to one that occurred in same regions of Venezuela and Colombia during 1962-1964. Symptoms of infected patients, estimated mortality rates, meteorological conditions preceding the epidemic, and seasonal patterns of transmission were all very similar to those reported in the previous outbreak. In addition, viruses isolated during 1995 were antigenically and genetically nearly identifical to those obtained during 1962-1964. These findings suggest that the epidemic resulted from the re-emergence of an epizootic serotype IC VEE virus. Identification of a similar virus isolate in mosquitoes in Venezuela in 1983, 10 years after epidemic/epizootic VEE activity ceased, raises the possibility of a serotype IC enzootic transmission cycle in northern Venezuela.

Adolescent↗
PubMed1995

Transmission of HIV in dialysis centre.

In August, 1993, 13 dialysis patients at one dialysis centre in Colombia, South America, were found to be HIV positive, and this prompted an epidemiological investigation. We carried out a cohort study of all dialysis centre patients during January, 1992 to December, 1993 (epidemic period) to determine risk factors for HIV seroconversion. Haemodialysis and medical records were reviewed, dialysis centre staff and surviving patients were interviewed, and dialysis practices were observed. Stored sera from all dialysis centre patients were tested for HIV antibody. 12 (52%) of 23 patients tested positive for HIV antibody by enzyme immunoassay and western blot during the epidemic period. Of the 23 tested, 9 (39%) converted from HIV antibody negative to positive (seroconverters) and 10 (44%) remained HIV negative (seronegatives). The HIV seroconversion rate was higher among patients dialysed at the centre while a new patient, who was HIV seropositive, was dialysed there (90% vs 0%; p < 0.01), or when the dialysis centre reprocessed access needles, dialysers, and bloodlines (60% vs 0%). While 2 of 9 HIV seroconverters had had sex with prostitutes, none had received unscreened blood products or had other HIV risk factors. No surgical or dental procedures were associated with HIV seroconversion. Dialysers were reprocessed separately with 5% formaldehyde and were labelled for use on the same patient. Access needles were reprocessed by soaking them in a common container with a low-level disinfectant, benzalkonium chloride; 4 pairs of needles were placed in one pan creating the potential for cross-contamination or use of one patient's needles on another patient. HIV transmission at the dialysis centre was confirmed. Improperly reprocessed patient-care equipment, most probably access needles, is the likely mechanism of transmission. This outbreak was discovered by accident and similar transmission may be occurring in many other countries where low-level disinfectants are used to sterilise critical patient-care equipment.

Adult↗
PubMed1995

Neutralization enzyme-linked immunosorbent assay for evaluation of immunity to measles virus.

A neutralization enzyme-linked immunosorbent (Nt-ELISA) assay for determination of protective immunity to measles virus was developed and evaluated. This procedure uses the same initial steps as performed to determine antibody titers by seroneutralization (Nt) test. However, a reduction in virus infectivity by neutralizing antibody was determined by quantitation of viral antigen using ELISA. The serum dilution that resulted in neutralization of 50% of infectious virus could be determined from the absorbance values. To be able to screen a large number of specimens, the conditions of the Nt-ELISA test were adjusted such that negative sera for measles antibodies and the positive ones were clearly distinguished on the basis of a single dilution (1:4). This test showed similar sensitivity (88.3%) and equal specificity as the Nt test when screening 136 serum samples from normal subjects. The estimation of protective antibody titers by Nt and Nt-ELISA methods was strongly correlated (correlation coefficient = 0.91). Thus, the measles Nt-ELISA test is rapid, reproducible, sensitive, and specific for detection of protective measles antibodies.

Antibodies, Viral↗
PubMed1993

Direct detection of wild poliovirus circulation by stool surveys of healthy children and analysis of community wastewater.

Cartagena, Colombia, was one of the last cities in the Americas known to have endemic poliomyelitis. After 3 cases were identified in 1991, two approaches for detecting continued silent transmission of wild polioviruses within a high-risk community were used: stool surveys of healthy children and virologic analysis of community sewage. Wild type 1 polioviruses were isolated from 8% of the children studied and from 21% of sewage samples. The proportions of wild polioviruses, vaccine-related polioviruses, and nonpolio enteric viruses were similar for both approaches. Wild poliovirus sequences were also amplified directly from processed sewage samples by the polymerase chain reaction using primer pairs specific for the indigenous type 1 genotype. The last reported cases associated with wild polioviruses in the Americas occurred in Colombia (8 April 1991) and Peru (23 August 1991). Direct sampling for wild polioviruses in high-risk communities can provide further evidence that eradication of the indigenous wild polioviruses has been achieved in the Americas.

Child, Preschool↗
PubMed1990

Distribution and etiology of leishmaniasis in Colombia.

A total of 340 Leishmania strains, isolated from humans, animals, and sand flies from various regions of Colombia, were examined by isozyme electrophoresis. Seven different Leishmania species were identified. Leishmania panamensis and L. braziliensis were the most common, representing 53.8% and 30.3% of the total, respectively. Isolation rates of the other species were as follows: L. chagasi, 9.4%; L. guyanensis, 2.6%; L. amazonensis, 1.8%; L. mexicana, 0.8%; and a new species requiring additional study, 1.2%. Statistical analyses of representative L. panamensis and L. braziliensis isolates indicated that the populations of these 2 species are genetically very similar. L. panamensis may have a continuous distribution in Colombia west of the eastern Andes Mountains and L. braziliensis may have a continuous distribution east of the western Andes Mountains. Information is given on disease manifestations of the parasites in human hosts and on isolation records from sand flies and animals.

Animals↗
PubMed1989

Epidemiology of visceral leishmaniasis in Colombia.

Epidemiologic studies were conducted during the period 1986-1988 in a small rural community in Colombia (El Callejon) where visceral leishmaniasis is highly endemic. In this community of 185 people, 14 cases of infantile visceral leishmaniasis were diagnosed in the 9 years 1981-1988. Leishmanin skin testing of a sample of the human residents showed that prevalence of Leishmania chagasi infection increased with age; overall, 51.2% of the subjects had a positive reaction. A canine surveillance program was instituted, using introduced sentinel dogs as well as the indigenous dog population. Eleven of 16 sentinel dogs were infected within 8 months of exposure; mean seroconversion time was 4.4 months. Eleven of 25 seronegative local dogs were also infected during the 26 month period; mean seroconversion time was 8 months. Parasites identified by isozyme electrophoresis as L. chagasi were recovered from 18 of 22 seropositive dogs. Collections of wild animals using baited live traps yielded mainly the neotropical opossum, Didelphis marsupialis. Leishmania chagasi was recovered from 12 of 37 (32.4%) opossums. Six of 681 female Lutzomyia longipalpis collected in the community had flagellates in their guts; cultures from 4 were identified as L. chagasi. These data confirmed that active parasite transmission occurred. The relatively high prevalence of L. chagasi infection found among D. marsupialis captured near human dwellings suggests that these animals may be an important peridomestic reservoir.

Age Factors↗
PubMed1989

Characterization of five new phleboviruses recently isolated from sand flies in tropical America.

Five new phlebotomus fever virus serotypes (Bunyaviridae: Phlebovirus) are described. These viruses, designated Ambe, Ixcanal, Mariquita, Armero, and Durania, were isolated from sand flies (Diptera: Psychodidae) collected in Brazil, Colombia, and Guatemala. Two of the agents were recovered from pools of male sand flies. The new viruses are antigenically related to other members of the phlebotomus fever serogroup by immunofluorescence, but are distinct from the other 39 members of this serogroup by plaque reduction neutralization test.

Animals↗
PubMed1989

AIDS in Colombia.

Between January 1984 and December 1987 a total of 178 AIDS cases were reported to the Colombian Ministry of Health. The location of these cases suggests that the human immunodeficiency virus (HIV) is widely distributed in Colombia. Most of those afflicted (97%) have been adult males. HIV seroprevalence studies of selected population groups revealed the highest antibody prevalence (5.65% in females, 22.5% in males) among individuals involved in high-risk behaviors who participated in a free AIDS testing program. High prevalences (from 0.6% to 3.9% in females, and 14.6% to 15.9% in males) were also found in patients (primarily female prostitutes and male homosexuals) attending clinics for sexually transmitted diseases in several urban areas. The number of AIDS cases in Colombia has doubled or tripled annually since reporting began in 1984, a pattern similar to that observed worldwide.

Acquired Immunodeficiency Syndrome↗
PubMed1987

Natural infection of humans, animals, and phlebotomine sand flies with the Alagoas serotype of vesicular stomatitis virus in Colombia.

Five isolations of the Alagoas serotype of vesicular stomatitis virus (Rhabdoviridae: Vesiculovirus) were made from naturally infected phlebotomine sand flies (Lutzomyia spp.) collected in Colombia. These are the first isolations of Alagoas virus from an arthropod. Replication of the virus occurred in laboratory-reared sand flies (Lutzomyia longipalpis) after inoculation. Bite and transovarial transmission of the virus was also demonstrated in experimentally infected sand flies. Alagoas virus neutralizing antibodies were found in sera of humans and animals living near the insect collection site; antibody rates among human residents of two nearby towns were 63% and 83%, respectively. Results of comparative serologic studies demonstrated that Alagoas virus is closely related antigenically to Indiana, Cocal, and Maraba viruses and that these four agents form a complex within the vesicular stomatitis virus serogroup. The antigenic similarity among these four viruses makes their differentiation difficult; it also raises doubts about the accuracy of current laboratory methods used for identifying isolates in this serogroup. A discussion follows on the significance of human antibodies to these agents and on the role of sand flies in their ecology.

Animals↗
PubMed1986

Biology of Arboledas virus, a new phlebotomus fever serogroup virus (Bunyaviridae: Phlebovirus) isolated from sand flies in Colombia.

Six isolates of a new phlebotomus fever serogroup virus, designated Arboledas virus, were obtained from sand flies (Lutzomyia spp.) collected in northeastern Colombia. One of the isolates was made from a pool of male sand flies. By immunofluorescence, Arboledas virus is related to Caimito and Pacui viruses; by neutralization test, it is distinct. Arboledas virus neutralizing antibodies were found in the sera of opossums (Didelphis marsupialis) and humans living in the study area. D. marsupialis inoculated with the virus developed a viremia of four days' duration, and sand flies (Lutzomyia gomezi) feeding on a viremic opossum were readily infected. Transovarial transmission of Arboledas virus was also demonstrated in experimentally infected Lu. gomezi. Results of the above laboratory studies suggest that Arboledas virus is maintained in nature by two mechanisms: vertical (transovarial) transmission in the insect vector, and an alternating marsupial-sand fly cycle. The implications of this complex maintenance cycle for other phleboviruses are discussed.

Animals↗