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Biomedical subjects

R S Nasci

Publications and source records attributed to R S Nasci.

At least 19 recordsLinked to original sources

Detection of eastern equine encephalitis virus in infected mosquitoes using a monoclonal antibody-based antigen-capture enzyme-linked immunosorbent assay.

Surveillance of mosquito populations for virus activity is not often performed by small, vector-control districts because they do not have the financial resources to use virus isolation, or newer methods such as the polymerase chain reaction. Consequently, development and refinements of rapid, sensitive, and simple enzyme-linked immunosorbent assays (ELISAs) applicable to a wide variety of public health settings are justified. We have developed an antigen-capture ELISA for the detection of eastern equine encephalitis (EEE) virus in mosquitoes that uses both monoclonal capture and detector antibodies. The sensitivity of this assay is 4.0-5.0 log10 plaque-forming units/ml, which is comparable to previously published EEE antigen-capture assays developed with polyclonal antibody reagents. This test identifies only North American strains of EEE virus and does not react with either western equine encephalitis or Highlands J viruses. Test sensitivity was enhanced by sonicating mosquito pools, treating them with Triton X-100, and increasing the time and temperature of antigen incubation. The conversion of this ELISA to a monoclonal antibody-based format should result in a readily standardizable and transferable assay that will permit laboratories lacking virus isolation facilities to conduct EEE virus surveillance.

Aedes↗

Rapid detection of west nile virus from human clinical specimens, field-collected mosquitoes, and avian samples by a TaqMan reverse transcriptase-PCR assay.

The authors report on the development and application of a rapid TaqMan assay for the detection of West Nile (WN) virus in a variety of human clinical specimens and field-collected specimens. Oligonucleotide primers and FAM- and TAMRA-labeled WN virus-specific probes were designed by using the nucleotide sequence of the New York 1999 WN virus isolate. The TaqMan assay was compared to a traditional reverse transcriptase (RT)-PCR assay and to virus isolation in Vero cells with a large number ( approximately 500) of specimens obtained from humans (serum, cerebrospinal fluid, and brain tissue), field-collected mosquitoes, and avian tissue samples. The TaqMan assay was specific for WN virus and demonstrated a greater sensitivity than the traditional RT-PCR method and correctly identified WN virus in 100% of the culture-positive mosquito pools and 98% of the culture-positive avian tissue samples. The assay should be of utility in the diagnostic laboratory to complement existing human diagnostic testing and as a tool to conduct WN virus surveillance in the United States.

Animals↗

La Crosse encephalitis virus habitat associations in Nicholas County, West Virginia.

Aedes triseriatus (Say) population density patterns and La Crosse encephalitis virus infection rates were evaluated in relation to a variety of habitat parameters over a 14-wk period. Ovitraps and landing collections were used in a La Crosse virus-enzootic area in Nicholas County, WV. Study sites were divided into categories by habitat type and by proximity to the residences of known La Crosse encephalitis cases. Results demonstrated that Ae. triseriatus population densities were higher in sugar maple/red maple habitats than in hemlock/mixed hardwood habitats or in a site characterized by a large number of small red maple trees. Sites containing artificial containers had higher population densities than those without. La Crosse virus minimum infection rates in mosquitoes collected as eggs ranged from 0.4/1,000 to 7.5/1,000 in the 12 study sites, but did not differ significantly among sites regardless of habitat type or proximity to human case residences. La Crosse virus infection rates in landing Ae. triseriatus mosquitoes ranged from 0.0/1,000 to 27.0/1,000. La Crosse virus was also isolated from host-seeking Ae. canadensis (Theobald) in two study sites, at rates similar to those found in the Ae. triseriatus populations. The Ae. triseriatus oviposition patterns and La Crosse virus infection rates suggest that this mosquito species disperses readily in the large woodlands of central West Virginia. The La Crosse enzootic habitats in Nicholas County, WV, are contrasted with those studied in other geographic regions where La Crosse virus is found.

Aedes↗

First field evidence for natural vertical transmission of West Nile virus in Culex univittatus complex mosquitoes from Rift Valley province, Kenya.

West Nile virus is a mosquito borne flavivirus endemic over a large geographic area including Africa, Asia, and the Middle East. Although the virus generally causes a mild, self-limiting febrile illness in humans, it has sporadically caused central nervous system infections during epidemics. An isolate of West Nile virus was obtained from a pool of four male Culex univittatus complex mosquitoes while we were conducting an investigation of Rift Valley fever along the Kenya-Uganda border in February-March 1998. This represents the first field isolation of West Nile virus from male mosquitoes and strongly suggests that vertical transmission of the virus occurs in the primary maintenance mosquito vector in Kenya. A phylogenetic analysis of the complete amino acid sequence of the viral envelope glycoprotein demonstrated a sister relationship with a Culex pipiens mosquito isolate from Romania made in 1996. This unexpected finding probably reflects the role of migratory birds in disseminating West Nile virus between Africa and Europe.

Amino Acid Sequence↗

Newly recognized focus of La Crosse encephalitis in Tennessee.

La Crosse virus is a mosquito-borne arbovirus that causes encephalitis in children. Only nine cases were reported in Tennessee during the 33-year period from 1964-1996. We investigated a cluster of La Crosse encephalitis cases in eastern Tennessee in 1997. Medical records of all suspected cases of La Crosse virus infection at a pediatric referral hospital were reviewed, and surveillance was enhanced in the region. Previous unreported cases were identified by surveying 20 hospitals in the surrounding 16 counties. Mosquito eggs were collected from five sites. Ten cases of La Crosse encephalitis were serologically confirmed. None of the patients had been discharged from hospitals in the region with diagnosed La Crosse encephalitis in the preceding 5 years. Aedes triseriatus and Aedes albopictus were collected at the case sites; none of the mosquitos had detectable La Crosse virus. This cluster may represent an extension of a recently identified endemic focus of La Crosse virus infection in West Virginia.

Adolescent↗

Epidemic leptospirosis associated with pulmonary hemorrhage-Nicaragua, 1995.

In October 1995, epidemic "hemorrhagic fever," without jaundice or renal manifestations, was reported in rural Nicaragua following heavy flooding; 2259 residents were evaluated for nonmalarial febrile illnesses (cumulative incidence, 6.1%) and 15 (0.7%) died with pulmonary hemorrhage. A case-control study found that case-patients were more likely than controls to have ever walked in creeks (matched odds ratio [MOR], 15.0; 95% confidence interval [CI], 1.7-132.3), have household rodents (MOR, 10.4; 95% CI, 1.1-97.1), or own dogs with titers >/=400 to Leptospira species (MOR, 23.4; 95% CI, 3.6-infinity). Twenty-six of 51 case-patients had serologic or postmortem evidence of acute leptospirosis. Leptospira species were isolated from case-patients and potential animal reservoirs. This leptospirosis epidemic likely resulted from exposure to flood waters contaminated by urine from infected animals, particularly dogs. Leptospirosis should be included in the differential diagnosis for nonmalarial febrile illness, particularly during periods of flooding or when pulmonary hemorrhage occurs.

Adolescent↗

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

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

Animals↗

Population size, parity structure, and wing length of Coquillettidia perturbans in an Ohio focus of eastern equine encephalitis.

Adult female density, parity status, and wing length were determined weekly for a population of Coquillettidia perturbans in an area enzootic for eastern equine encephalitis virus in central Ohio. Samples were collected in CO2-baited CDC miniature light traps from the first week in June through the 2nd week of September 1992. Population density indicated a single emergence peak during the 2nd week in July. However, parity rates showed 2 peaks, occurring in the first week of August (70.9% parous) and the 2nd week of September (55.3% parous), which suggested that there was a relatively small 2nd generation. Average wing length declined significantly over the season. The decline in size was negatively correlated with average air temperature occurring at least 6 wk before the time of emergence. Despite the seasonal decline in wing length, the low coefficient of variation for the average wing length (5.5) indicated relatively little variation in size. Comparison of parous and nulliparous female wing lengths each week suggested that there was no association between size and survival in this species.

Animals↗

Arbovirus titer variation in field-collected mosquitoes.

Patterns in the distribution of titers in arbovirus-positive mosquito pools were examined. Virus isolation records from the Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, from 1974 through 1993 were used to estimate virus titers in field-collected pools. Pools were classified as either low titer (< or = 3.0 log10 plaque-forming units [PFU]/ml) or high titer (> 3.0 log10 PFU/ml). The proportion of virus-positive pools that had high titers varied among the different domestic arboviruses, within viruses among field sites and years, and within viruses among mosquito species tested. Alphaviruses produced a greater proportion of pools with high titers than did the flaviviruses. Variation in the proportion of pools with high titers among sites and years suggested variation in mosquito and/or virus strains. Variation in the proportion of pools with high titers among mosquito species indicated species-specific differences in vectorial capacity. The results show that information about the titer of virus in mosquito pools can complement other parameters, such as the minimum infection rate, currently used in mosquito-based arbovirus surveillance programs.

Animals↗

Control of Aedes albopictus from the perspective of North America.

Aedes albopictus is a serious nuisance species and a potential vector of domestic arboviruses in the U.S. Public health and mosquito control agencies have taken numerous actions to address the problems posed by this species. Federally mandated programs of disinsection and inspection of tire casings imported into the U.S. are aimed at preventing additional introductions of this and other exotic mosquito species. Control programs, including the use of traditional chemical larvicides and adulticides, as well as the introduction of predatory copepods into larval habitats, have been developed and tested for population management. A few of the established populations in the northern areas of Ae. albopictus distribution in the continental U.S. have been successfully eradicated through source reduction and insecticide application.

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Larval diet, adult size, and susceptibility of Aedes aegypti (Diptera: Culicidae) to infection with Ross River virus.

The relationship of larval nutrition and adult body size to the susceptibility of Aedes aegypti (L.) to Ross River virus infection was examined. Large adult mosquitoes produced by feeding larvae a high-level diet consumed significantly more virus particles than did smaller mosquitoes. However, when a correction for body size was made, smaller mosquitoes were found to consume significantly more virus per unit of body weight. A host viremia of 2.4 log10 PFU/ml failed to infect mosquitoes of any size. Large Ae. aegypti mosquitoes were significantly more susceptible than small mosquitoes when fed on hosts with viremias of 4.8, 5.7, 6.4, and 7.5 log10 PFU/ml, but differences in susceptibility were less apparent at higher viremias.

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Aedes albopictus (Diptera: Culicidae) population density and structure in southwest Louisiana.

Pupal density, wing length at emergence, host-seeking female abundance, and host-seeking female wing length and parity were determined monthly for a population of Aedes albopictus (Skuse) in a southwestern Louisiana tire dump from January 1988 through January 1989. Pupae and host-seeking females were first collected in April. Pupal densities were consistently high from June through September. Host-seeking female abundance was highest during July and August. Average female wing length was longest during spring, declined in midsummer, and increased slightly during fall. Average wing length was negatively correlated with average pupal density during the month of collection and with average air temperature recorded the previous month. Host-seeking females had consistently longer wings than emerging females, and parous host-seeking females had longer wings than nulliparous host-seeking females. Average wing length of the parous host-seeking females was consistently long throughout the season. Parity of host-seeking females parity was highest during July when approximately 21 parous host-seeking females were collected per hour. These results indicate that seasonal patterns in immature population density influence average body size and parity structure in Ae. albopictus.

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Control of Aedes albopictus larvae using time-release larvicide formulations in Louisiana.

The ability of time-release formulations of larvicides and insect growth regulators (IGRs) to provide long-term control of Aedes albopictus was investigated in the field. Larvicides used in the study were Bactimos pellets (Bacillus thuringiensis var. israelensis, active ingredient) and Abate pellets (temephos, active ingredient). The IGR Altosid (methoprene, active ingredient) was used in pellet and sand formulations. Application rates were higher than label recommendations. In a preliminary test, clay flower bowls were treated with 2 g of material. Bactimos pellets failed to provide control after 60 days. Abate pellets and the Altosid formulations provided essentially 100% control for 150 days. After 360 days in the field, the Abate pellets produced 100% larval mortality, and significant levels of control were provided by the Altosid formulations and the Bactimos pellets. In a small-scale operational trial of this technique, 1 g of Altosid pellets was applied to every container that could be located in 2 urban residential neighborhoods in Lake Charles, LA. Aedes albopictus biting populations were monitored weekly in the treated areas and in an untreated control area. Biting population densities declined significantly in treated areas compared with the control area. Results suggested that long-term control of Ae. albopictus populations can be achieved economically with one application of Altosid pellets or Abate pellets in containers.

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Blood hosts of Aedes albopictus in the United States.

Bloodfed Aedes albopictus were collected during 1989-91 by vacuum aspirator from rural and urban study sites in Missouri, Florida, Indiana, Illinois, and Louisiana. Blood hosts identified by ELISA and precipitin tests were rabbit (n = 91), Rattus sp. (n = 69), dog (n = 14), unidentified mammal (n = 14), cow (n = 13), human (n = 10), deer (n = 10), sciurid (n = 7), turtle (n = 5), murid other than Rattus sp. (n = 4), raccoon (n = 3), passeriform bird (n = 3), and cat (n = 2). As an opportunistic bloodfeeder, Ae. albopictus may be a potential vector of domestic arboviruses and a nuisance pest where infestations occur.

Aedes↗

Eastern equine encephalitis virus in Ohio during 1991.

During August and September of 1991, an epizootic of eastern equine encephalitis (EEE) virus in horses occurred in Wayne and Holmes countries, OH. This was the first recorded epizootic of EEE virus in the state. Twelve horses were confirmed positive for EEE virus through virus isolation or seroconversion, and seven additional horses with compatible symptoms were in close spatial and temporal proximity to the confirmed cases and were presumed to have died from EEE virus. The outbreak was centered around the Killbuck Wildlife Area, a 2,147-ha tract maintained by the state, half of which consists of wooded swamp and marsh. Mosquitoes were collected in upland areas before the epizootic and in the swamp basin at the end of the epizootic to identify the mosquito species involved in EEE virus transmission. We collected and tested 22,095 specimens for the presence of virus. EEE virus was isolated from one pool of the most likely epizootic vector, Coquillettidia perturbans (Walker). The minimum infection rate for EEE virus in this species was 0.1/1,000. Dense populations of Aedes vexans (Meigen) and Culex salinarius Coquillett occurred in the area, but their densities peaked after the epizootic. It is unlikely that these species were involved in epizootic transmission. IgM antibody to EEE virus was detected in three bird species collected in the swamp.

Animals↗