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Biomedical subjects

J L Hardy

Publications and source records attributed to J L Hardy.

At least 37 records · Page 2Linked to original sources

Natural vertical transmission of western equine encephalomyelitis virus in mosquitoes.

The mechanism by which western equine encephalomyelitis (WEE) virus and other mosquito-borne alphaviruses (Togaviridae) survive during periods of vector inactivity is unknown. Recently, three strains of WEE virus were isolated from adult Aedes dorsalis collected as larvae from a salt marsh in a coastal region of California. This provides evidence of vertical transmission of WEE virus in mosquitoes in nature. Vertical transmission in Ae. dorsalis and closely related mosquito species may be an important mechanism for the maintenance of WEE virus in temperate regions in North America where horizontal transmission of the virus is seasonal.

Aedes↗

Potential effect of global warming on mosquito-borne arboviruses.

If global warming occurs in California, daily mean temperatures may increase by 3 to 5 degrees C, precipitation patterns will change, and sea level may rise 1 m. Studies were done on effect of temperature changes on survival of Culex tarsalis Coquillett, the primary vector of western equine encephalomyelitis (WEE) and St. Louis encephalitis (SLE) viruses, in two regions where temperatures differed by 5 degrees C. Daily mortality of adult vectors increased by 1% for each 1 degree C increase in temperature. At 25 degrees C, only 5% of Cx. tarsalis survived for 8 or more days, the time required for extrinsic incubation of these viruses. Extrinsic incubation times for these viruses shortened when temperatures were increased from 18 to 25 degrees C. WEE virus infection was modulated and transmission decreased at 32 degrees C. If temperatures in the warmer region increase by 5 degrees C, WEE virus may disappear and SLE virus would persist. In the cooler region, a 5 degrees C increase would decrease vector survivorship and virus activity in midsummer. In North America, epidemics of WEE have prevailed above a 21 degrees C isotherm and those of SLE below this isotherm. With global warming, epidemics of these viruses could extend into currently unreceptive northern areas. WEE virus would disappear from more southern regions. Geographic distribution of vector, human, and animal populations could be altered. North America could become more receptive to invasion by tropical vectors and diseases.

Animals↗

Viremia and serological responses in adult chickens infected with western equine encephalomyelitis and St. Louis encephalitis viruses.

Adult hens, similar to those used for arbovirus surveillance, were experimentally infected with western equine encephalomyelitis (WEE) and St. Louis encephalitis (SLE) viruses to describe the viremia response, to compare serological testing methods, and to evaluate a new method of collecting whole blood onto filter paper strips from lancet pricks of the chicken comb. Young (19 weeks), but not old (38 weeks), hens developed a low-titer, transient viremia for a 1-day period. Immunoglobulin G (IgG) was detected by days 10 and 14 after infection with WEE and SLE viruses, respectively, by indirect fluorescent antibody tests, hemagglutination inhibition tests, and plaque reduction neutralization tests on sera and in direct enzyme immunoassays (EIA) on both sera and eluates from filter paper samples. Immunoglobulin M (IgM) was first detected in sera 2 and 3 days before IgG, respectively, but IgM could not be detected reliably in eluates from dried blood. Sera and dried blood samples collected from naturally infected sentinel chickens gave comparable results when tested by an EIA for IgG.

Animals↗

Effect of temperature on the transmission of western equine encephalomyelitis and St. Louis encephalitis viruses by Culex tarsalis (Diptera: Culicidae).

The extrinsic incubation rate (inverse of the time in days from infection to median transmission) of western equine encephalomyelitis (WEE) and St. Louis encephalitis (SLE) viruses by laboratory strains of Culex tarsalis Coquillett increased as a linear function of incubation temperatures from 10 to 30 degrees C. The estimated temperatures for zero transmission thresholds (intercept of the X axis) were 10.9 and 14.9 degrees C, and the number of degree days above these thresholds required for median transmission (inverse of the slope) was 67.6 and 115.2, respectively. Although the bodies of most Cx. tarsalis females remained infected and the WEE viral titer did not decrease significantly throughout the incubation periods at all temperatures, transmission rates by females incubated at 20 to 30 degrees C decreased markedly after peaking at 7-10 d after infection. In contrast, midgut escape and salivary gland infection barriers limited the transmission rates of SLE virus at all temperatures, but these rates did not decrease markedly as a function of incubation time, indicating that virus modulation did not occur. Degree-day models were used to calculate monthly changes in the duration of the extrinsic incubation period for WEE and SLE viruses in the San Joaquin and Coachella valleys based on mosquito temperatures estimated by combining nocturnal air and diurnal resting site temperatures. Temperatures in the San Joaquin Valley averaged 5 degrees C cooler than in the Coachella Valley, proportionately shortening the duration of the potential transmission season for WEE virus from 10 to 8 mo and for SLE virus from 8 to 5 mo, respectively.

Animals↗

Vector competence of alpine, Central Valley, and coastal mosquitoes (Diptera: Culicidae) from California for Jamestown Canyon virus.

Mosquitoes collected from alpine, Central Valley, and coastal habitats in California were evaluated for their vector competence for four strains of Jamestown Canyon (JC) virus. Three of the viral strains examined were isolated from alpine Aedes species collected in California, and one, the prototype JC virus, was isolated from Culiseta inornata (Williston) collected in Colorado. Alpine Aedes tahoensis Dyar, Ae. cataphylla Dyar, Ae. hexodontus Dyar, Ae. increpitus Dyar, Ae. clivis Lanzaro and Eldridge, and coastal Aedes washinoi Lanzaro and Eldridge were variably susceptible to alpine strains of JC virus. Infection rates ranged from 22 to 77%, and peroral transmission rates of the infected females ranged from 0 to 26%. The differences were related to both mosquito species and viral strain. Coastal populations of Cs. inornata, Ae. washinoi, and Ae. sierrensis (Ludlow) were incompetent vectors when fed an alpine strain of JC virus, whereas Ae. squamiger (Coquillet) and Ae. dorsalis (Meigen) were competent vectors. Peroral transmission rates following parenteral infection of females of most species were about twofold higher than those for perorally infected females. A population of Cs. inornata from the Central Valley was highly susceptible when fed an alpine strain of JCV and transmitted virus both horizontally and vertically. Alpine strains of JC virus also were transmitted vertically by Ae. tahoensis, Ae. washinoi, and Ae. squamiger following parenteral infection of females.

Aedes↗

Vector competence of selected mosquito species (Diptera: Culicidae) for California strains of Northway virus (Bunyaviridae: Bunyavirus).

Selected mosquito species from Central Valley, coastal, and alpine habitats of California were evaluated for their vector competence for Northway (NOR) virus. Culiseta incidens Thomson, Culiseta inornata (Williston), and Anopheles freeborni Aitken were the only competent vectors when fed virus. Aedes sierrensis (Ludlow), as well as alpine snow pool Aedes (i.e., Ae. cataphylla Dyar, Ae. hexodontus Dyar, Ae. increpitus Dyar and Ae. tahoensis Dyar), Ae. melanimon Dyar, Ae. washinoi Lanzaro & Eldridge, Culex erythrothorax Dyar, and Cx. tarsalis Coquillett were highly refractory to peroral infection. Alpine snow pool species were poor vectors even following parenteral infection, but 21-35% of intrathoracically inoculated Ae. sierrensis from several lower elevation localities and 39-46% of parenterally infected Cx. tarsalis were able to transmit NOR virus per os. Perorally infected Cs. inornata transmitted NOR virus vertically to their F1 adult progeny at a low rate (minimal filial infection rate, 1:248).

Animals↗

Isolations of Jamestown canyon virus (Bunyaviridae: California serogroup) from mosquitoes (Diptera: Culicidae) in the western United States, 1990-1992.

Nearly 80,000 immature and adult mosquitoes in three genera were collected in high-elevation (> 1,000 m) areas of California (68,229), Nevada (3,721), Oregon (5,918), and Washington (1,629) during 1990-1992 and tested for virus as adult males or females in 1,799 pools. Collections comprised primarily alpine Aedes in the Aedes communis (De Geer) group of the subgenus Ochlerotatus. Thirteen strains of Jamestown Canyon (JC) virus were recovered by plaque assay in Vero cell culture from three members of the Ae. communis group: 10 from Aedes tahoensis Dyar, 2 from Aedes cataphylla Dyar, and 1 from Aedes hexodontus Dyar. All isolates came from collections made in Alpine, Sierra, Tulare, or Tuolumne counties in the Sierra Nevada of California. Vertical transmission of JC virus in all three mosquito species was demonstrated by the isolation of virus from adult males or females reared from field-collected larvae or pupae. The prevalence of infected Ae. tahoensis was significantly higher in field-collected adult females than in reared adult males and females in Alpine County, which indicated that JC virus was being amplified by horizontal transmission. This study further incriminated Ae. tahoensis, Ae. cataphylla, and Ae. hexodontus as natural vectors of JC virus in California and greatly extended the known geographical range of this virus in the Sierra Nevada.

Altitude↗

Seroepidemiology of California and Bunyamwera serogroup bunyavirus infections in humans in California.

Several human populations in California were surveyed cross-sectionally and longitudinally for neutralizing antibodies to selected arthropod-borne bunyaviruses in the California and Bunyamwera serogroups. Overall, the prevalence of antibodies to California serogroup viruses was 6.4% in 702 individuals sampled during 1963-1988. Comparative antibody titers in individual sera indicated that 4.1% and 1.6% of these infections were caused by viruses similar or identical to Jamestown Canyon and California encephalitis, respectively. Evidence of prior infection with the Jamestown Canyon serotype was found in 10% of 118 humans employed outdoors in high elevation areas and sampled in 1988, including 5 of 16 persons (31%) employed as rangers patrolling in remote forests and meadows. This probably reflects increased exposure to bites of boreal mosquitoes that breed in pools of melted snow. Antibodies to Bunyamwera serogroup viruses, including the Northway serotype, which was recently shown to be enzootic in California, were found in only 2 of 702 humans studied. No seroconversions were detected to selected California or Bunyamwera serogroup viruses in paired samples from 392 humans, including 349 patients with acute central nervous system disease or undifferentiated febrile illnesses who were sampled during 1963-1988, and thus these viruses are currently unconfirmed as human pathogens in California.

Adolescent↗

Ecological observations on the 1989 outbreak of St. Louis encephalitis virus in the southern San Joaquin Valley of California.

Temporal and spatial patterns of St. Louis encephalitis (SLE) virus transmission were compared at permanent study areas in the southern San Joaquin Valley during years with low (1988 and 1990) and elevated (1989) viral activity. During 1989 and 1990, virus appeared first at sentinel chicken flocks exhibiting low to moderate seroconversion rates at the end of the previous season. This finding, and the early season seroconversion of sentinel chickens at a marsh habitat on 5 March and 2 April 1990, circumstantially indicated that SLE virus may have overwintered on the valley during the winters of 1988-1989 and 1989-1990. The mechanism of overwintering was not elucidated further, because virus could not be isolated from overwintering adult mosquitoes or from immatures collected during the spring. An outbreak of 26 confirmed SLE cases occurred in 1989 during a drought year (rainfall 50% of normal) and followed a spring with elevated temperatures (1.7-3.4 degrees C above normal) and Culex tarsalis Coquillett abundance. Cx. tarsalis was the primary vector, being most abundant during the virus amplification period in early summer and most frequently infected (70 SLE virus positive pools/329 tested). SLE virus also was detected in Culex quinquefasciatus Say (14/65) and Cx. stigmatosoma Dyar (1/4); however, both species were distributed focally and increased in abundance only after widespread seroconversions had occurred in sentinel chickens. Increased virus activity during 1989 was not accompanied by marked changes in vector susceptibility or in SLE virus infectivity for mosquitoes. Decreased virus activity in the Bakersfield area during 1990 could not be attributed to immunity in passeriform birds, because a small seroprevalence survey indicated that few adult birds had antibodies to SLE virus.

Animals↗

Mosquito and arbovirus ecology in southeastern California, 1986-1990.

Mosquito abundance and western equine encephalomyelitis (WEE) and St. Louis encephalitis (SLE) virus activity were monitored in five valleys in southeastern California from June 1986 through April 1990 to study virus overwintering and possible dissemination from south to north along geographically defined corridors. Culex tarsalis Coquillett predominated in CO2 trap collections and was the only species repeatedly infected with WEE and SLE viruses. Abundance peaked during April-May and August-October. WEE virus infections in Cx. tarsalis generally were detected after the spring peak and were followed approximately 1 mo later by seroconversions in sentinel chickens. SLE virus infections occurred later in the summer but before the fall peak in Cx. tarsalis abundance. Peak Cx. tarsalis abundance occurred when monthly temperatures averaged 25 degrees C, whereas virus infections generally were detected most frequently when temperatures exceeded 29 degrees C. Although the spring increase in Cx. tarsalis abundance occurred earlier in southern valleys, the onset of virus activity was variable among valleys and did not follow a south to north progression.

Animals↗

Ecology of mosquitoes and St. Louis encephalitis virus in the Los Angeles Basin of California, 1987-1990.

St. Louis encephalitis (SLE) virus has become established in the Los Angeles Basin of California, where it most likely is maintained by horizontal transmission among Culex tarsalis Coquillet and passerine birds in park-riparian habitat. Viral transmission also was detected at low levels in residential habitat by the infrequent seroconversion of sentinel chickens. SLE virus activity was documented in all months except March and April. Cx. tarsalis was incriminated as the primary vector, based on elevated field infection rates (0.17 infected females per 1,000 tested), population abundance concomitant in time and space with increased SLE virus activity, vector competence, and vectorial capacity. Culex stigmatosoma Dyar also may be important in the horizontal maintenance of SLE virus because this species was an efficient laboratory vector, fed frequently on passeriform birds, and had field infection rates (0.15 females per 1,000) comparable with Cx. tarsalis. However, difficulty in trapping adult females precluded the assessment of Cx. stigmatosoma because abundance may have been underestimated and comparatively few females were tested for virus. Although Cx. quinquefasciatus Say was the most abundant species in residential habitats (greater than 85% of total catch at CO2 traps) where it occasionally fed on humans (0.6% of total tested), this species was considered to be of secondary importance because field infection rates were low (0.03 females per 1,000) and few females became infected and transmitted SLE virus in laboratory vector competence experiments. Although Culex erythrothorax Dyar fed on humans in marsh habitat (5% of total tested), this species infrequently fed on birds (12%) and was refractory to SLE virus infection in vector competence experiments; therefore, it was not considered a vector of SLE virus. Future SLE virus surveillance and mosquito control programs should be directed toward Cx. tarsalis populations.

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Evaluation of domestic pigeons as sentinels for detecting arbovirus activity in southern California.

Flocks of sentinel domestic pigeons (Columbia livia) detected increases in St. Louis encephalitis (SLE) and western equine encephalomyelitis (WEE) virus activity in southern California concurrently with flocks of sentinel chickens. However, occasional low-titered, transient seroconversions to both WEE and SLE viruses also occurred in pigeons during periods when virus activity was not detected by seroconversions in sentinel chickens, by virus isolation from Culex mosquitoes, or by human disease. Moreover, SLE virus seroconversions detected in pigeons by a hemagglutination inhibition (HI) test frequently could not be confirmed either by a plaque-reduction neutralization test (PRNT) on the same sera, or by an HI test on the next monthly serum sample from the same bird. Experimental infection studies, in which pigeons were inoculated subcutaneously with SLE (SOUE 16-84) virus, confirmed that pigeons developed low-titered and transient HI antibodies that were detectable infrequently by PRNT. In contrast, experimental infection with WEE (BFS 1703) virus produced elevated antibody responses that were detectable by HI for 8-12 weeks and by PRNT for at least 25 weeks. Pigeons infected with SLE virus rarely developed detectable viremias, whereas most birds infected with WEE virus developed viremias on postinfection day 1 that persisted for two or three days. Host-preference studies indicated that pigeons were less attractive as bait in lard can traps to host-seeking Culex mosquitoes than were chickens, and that blood-engorged Culex females collected near sentinel locations fed more frequently upon galliform than columbiform birds. Collectively, these results indicated that sentinel pigeons would not provide an adequate replacement for sentinel chickens to monitor WEE or SLE viruses, and would be a dead-end host for SLE virus.

Animals↗

Vector competence of California mosquitoes for California encephalitis and California encephalitis-like viruses.

Mosquitoes collected from coastal, inland valley, and alpine locations in California were evaluated for their experimental vector competence for two viruses in the California serogroup (Bunyaviridae:Bunyavirus). Aedes squamiger, a coastal salt marsh mosquito, was an efficient vector of a California encephalitis (CE)-like virus isolated from its habitat (89% of the pledget-fed females became infected and 61% transmitted virus). Aedes dorsalis, a coastal mosquito, and Ae. melanimon, an inland valley mosquito, were competent vectors of prototype CE virus (98% and 100% of the pledget-fed females became infected and 56% and 30%, respectively, transmitted virus). Aedes squamiger and Ae. dorsalis transmitted both viruses vertically to one or more of 20 of their progeny. Culiseta inornata was susceptible to infection with both viruses, but 5% or less transmitted virus perorally. Alpine mosquitoes, Ae. cataphylla, Ae. increpitus, and Ae. tahoensis, became infected with both CE and CE-like viruses, but 3% or less transmitted virus. All species of mosquitoes were more efficient vectors of both viruses following intrathoracic inoculation than following pledget feeding, suggesting the presence of mesenteronal barriers.

Animals↗

Occurrence and evolutionary significance of a California encephalitis-like virus in Aedes squamiger (Diptera: Culicidae).

More than 12,000 Aedes increpitus Dyar and 4,600 Aedes squamiger (Coquillett) were tested for the presence of arboviruses to test the hypothesis that there is a coevolutionary relationship between Aedes (Ochlerotatus) mosquitoes and California serogroup viruses. Five strains of a California encephalitis-like virus were isolated from adults reared from larvae of Ae. squamiger collected in January 1989 from a coastal salt marsh at Morro Bay, San Luis Obispo County, California. Viruses were isolated in Vero cell cultures and serotyped by cross-neutralization tests. These isolates represent the first arboviruses isolated from this species. On the basis of morphology, Aedes squamiger has been included in the Aedes stimulans group of the subgenus Ochlerotatus. Other species within the Ae. stimulans group are vectors of California (CAL) serogroup viruses elsewhere in North America. Analysis of isozyme variability supports the inclusion of Ae. squamiger in the Ae. stimulans group and suggests that coastal populations of Ae. increpitus are the closest California relatives of Ae. squamiger. Recovery of virus from Ae. squamiger reinforces the relationship between CAL serogroup viruses and Aedes (Ocherlotatus) mosquitoes. However, the failure to isolate virus from large samples of Ae. increpitus from coastal and low elevation inland habitats suggests a complex evolutionary history involving both vertical and horizontal transmission mechanisms.

Aedes↗

Isolation of Jamestown Canyon virus from boreal Aedes mosquitoes from the Sierra Nevada of California.

More than 28,000 mosquitoes in four genera were collected from high elevation (greater than or equal to 1,000 m) areas of California during 1988-89 and tested for virus by plaque assay in Vero cells. Viruses were serogrouped by enzyme immunoassay and serotyped by cross-neutralization. Six strains of Jamestown Canyon virus in the California serogroup were isolated from three species of boreal Aedes in the Aedes communis group of the subgenus Ochlerotatus. All isolates were from mosquitoes collected in Alpine County at approximately 2,300 m elevation in the Sierra Nevada. These included one virus from a pool of male Aedes cataphylla collected in immature stages, which is evidence for vertical transmission; four viruses from adult female Ae. communis (sens. lat.); and one virus from adult female Aedes hexodontus. These are the first isolations of viruses from boreal Aedes mosquitoes in California and the first reported isolations of Jamestown Canyon virus from Ae. cataphylla or Ae. hexodontus.

Aedes↗

Isolation of Northway serotype and other Bunyamwera serogroup bunyaviruses from California and Oregon mosquitoes, 1969-1985.

Eight previously untyped Bunyamwera serogroup bunyaviruses that had been isolated from mosquitoes collected in California and Oregon between 1969 and 1985, were identified by cross-neutralization tests. Four viruses from Anopheles freeborni and a virus from Aedes sierrensis collected in Butte County in the Central Valley of California in 1970-71 were shown to belong to the Northway serotype. The existence of a Northway serotype virus in California had been inferred from previous serologic surveys of deer and horses, but these are the first Northway serotype viruses to be identified from the contiguous United States or from An. freeborni. This is also the first isolation of any virus from Ae. sierrensis. Two viruses from Culiseta inornata collected in Umatilla County, Oregon in 1969 may represent a new serotype in the Bunyamwera serocomplex of the Bunyamwera serogroup. The name "Stanfield" is proposed for this serotype, which is closely related to the Cache Valley and Northway bunyavirus serotypes. A virus from Ae. taeniorhynchus collected in San Diego County, California in 1985 was confirmed to belong to the Main Drain serotype. Previously, Main Drain serotype viruses in California have been associated principally with Culicoides variipennis.

Aedes↗

Binding of western equine encephalomyelitis virus to brush border fragments isolated from mesenteronal epithelial cells of mosquitoes.

Brush border fragments (BBF) were isolated from mesenteronal epithelial cells of mosquitoes that are either susceptible (WS Culex tarsalis) or refractory (WR Cx. tarsalis; Culex pipiens) to peroral infection by western equine encephalomyelitis (WEE) virus. The isolated BBF were combined with radiolabeled WEE virus in a binding assay to compare the amount of virus bound by BBF from susceptible and refractory mosquitoes. BBF and WEE virus were mixed in a microcentrifuge tube, incubated for 1 h and centrifuged at 27,000 X g-30 min to pellet WEE virus bound to BBF. Optimal binding occurred at pH 7.2, 20 degrees C and there was no requirement for divalent cations. BBF isolated from perorally susceptible mosquitoes (WS Cx. tarsalis) bound significantly greater amounts of radiolabeled WEE virus, compared to BBF isolated from refractory mosquitoes (WR Cx. tarsalis; Cx. pipiens), in all experiments. The binding of WEE virus to BBF from WS Cx. tarsalis appears to be specific, based on saturation and competitive binding studies; binding to BBF from WR Cx. tarsalis and Cx. pipiens is nonspecific. Scatchard analysis of the binding data for BBF from WS Cx. tarsalis yields an estimated 1.8-3.5 X 10(6) binding sites per mesenteronal epithelial cell with an affinity constant of (Ka) of 2.2 X 10(11) M-1.

Animals↗

Diel changes in adult mosquito microhabitat temperatures and their relationship to the extrinsic incubation of arboviruses in mosquitoes in Kern County, California.

Microhabitat temperatures experienced by nocturnally active Culex tarsalis Coquillett mosquitoes were measured in the southern San Joaquin Valley of California. Temperatures of the diurnal resting shelter and nocturnal activity air space were measured by digital recorder at three Cx. tarsalis habitats in Kern County. Temperatures measured by digital recorders agreed well with temperatures recorded concurrently at nearby California Irrigation Management and Inspection System weather stations. Monthly temperatures among habitats were combined to depict the diel temperature regimen experienced by female Cx. tarsalis as a consequence of daily movement between the diurnal resting shelter and nocturnal air space. Transitions between the two microhabitats corresponded to the times of mosquito ingress and egress (i.e., sunrise and sunset). The composite daily temperature mean ranged from 7.4 degrees C in December to 21.5 degrees C in July, and temperature extremes seldom exceeded a daily maximum of 25.0 degrees C during the summer or dropped below a daily minimum of 5.0 degrees C during the winter. Overall, mosquitoes occupied a composite thermal environment where the extrinsic incubation of an arbovirus would have occurred at ambient temperatures that were cooler than either the diurnal resting shelter or activity air space.

Animals↗