High Seroprevalence of Jamestown Canyon Virus among Deer and Humans, Nova Scotia, Canada.

Using residual serum samples from Nova Scotia, Canada, we found that 87.8% of tested deer and an estimated 20.6% of the human population were infected with Jamestown Canyon virus. Human seropositivity reached 48.2% in 1 region. This virus may be an underrecognized cause of disease in Nova Scotia.

Risk factors associated with seropositivity to California serogroup viruses in humans and pet dogs, Quebec, Canada.

Jamestown Canyon and snowshoe hare viruses are two emerging human pathogens associated with cases of neuroinvasive disease in North America. This study aimed to identify environmental and individual risk factors for seropositivity to these arboviruses in humans and pet dogs from Québec, Canada, 2012-2014. In humans, areas with moderate densities of white-tailed deer (Odocoileus virginianus) were associated with higher odds of seropositivity compared with areas with low densities of white-tailed deer (OR 2.50, P = 0.009) and odds of seropositivity were higher in males than in females (OR 2.03, P = 0.016). Among humans reporting more than 10 mosquito bites weekly, the odds of being seropositive were 4.44 times higher (P = 0.004) for people living in hardwood forested areas. Exposure to areas with coniferous forests was identified as the main environmental risk factor for seroconversion in dogs (OR 2.39, P = 0.04). These findings may help target further public health research, diagnostic and surveillance efforts in Canada.

Roles of host species, geographic separation, and isolation in the seroprevalence of Jamestown Canyon and snowshoe hare viruses in Newfoundland.

California serogroup viruses, including Jamestown Canyon virus (JCV) and snowshoe hare virus (SSHV), are mosquito-borne members of the Bunyaviridae family and are endemic across North America. These arboviruses are potential pathogens which occasionally cause neuroinvasive disease in humans and livestock. A neutralization assay was used to document JCV and SSHV seroprevalence using blood collected from a variety of domestic and wildlife host species. These species were sampled in an island setting, Newfoundland, which contains diverse ecoregions, ecological landscapes, and habitats. Seroprevalence rates for each virus differed significantly among host species and within certain species across different geographic areas. JCV was significantly associated with large mammals, and SSHV was significantly associated with snowshoe hares. Seroprevalence rates in the 5 species of animals tested for prior exposure to JCV ranged from 0% in snowshoe hares to 64% in horses. Seroprevalence rates for SSHV ranged from less than 1% in bovines to 55% in all snowshoe hares. The seroprevalence of SSHV differed significantly (P < 0.05) among hares occupying the discrete habitats of watersheds separated by 14 to 35 km. Cattle on farms in boreal forest landscapes displayed significantly higher JCV seroprevalence (P < 0.001) than those on farms located in seacoast landscapes. Lifelong geographic isolation of cattle to insular Newfoundland was associated with significantly lower JCV seroprevalence (P < 0.01) than that for cattle which had lived off-island.

Carbon dioxide sensitivity of mosquitoes infected with California encephalitis virus.

Four species of mosquitoes became sensitive to carbon dioxide approximately 3 to 4 days after they received intrathoracic injectins of California encephalitis virus. Aedes melanimon and Aedes dorsalis infected orally with California encephalitis virus also became carbon dioxide-sensitive, but mosquitoes infected transovarially did not. Sensitivity to carbon dioxide was inhibited by antiserum to California encephalitis virus. To our knowledge this is the first report of carbon dioxide sensitivity induced in arthropods by a bunyavirus and the first demonstration of this phenomenon by an arbovirus in its proven vector.

Serological surveillance for Tahyna virus (California encephalitis orthobunyavirus, Peribunyaviridae) neutralizing antibodies in wild ungulates in Austria, Hungary and Romania.

A serosurvey for Tahyna virus (TAHV), a mosquito-borne California encephalitis orthobunyavirus (Peribunyaviridae) endemic to Europe, was performed to estimate the activity of TAHV on a broad geographic scale. Sera from wild boar (Sus scrofa), roe deer (Capreolus capreolus) and red deer (Cervus elaphus) were collected from Austria, Hungary and Romania. Samples were tested for neutralizing antibodies against TAHV using a virus microneutralization assay. The results demonstrate that TAHV transmission to mammals is widespread in Europe, particularly in the wild boar population where the mean rate of seroconversion is 15.2%.

Experimental infection of chipmunks and snowshoe hares with La Crosse and snowshoe hare viruses and four of their reassortants.

The infection of chipmunks (Tamias striatus), the vertebrate host of La Crosse (LAC) arbovirus, or snowshoe hares (Lepus americanus), the host of snowshoe hare (SSH) virus, was analyzed following subcutaneous inoculation with either LAC or SSH or certain LAC-SSH reassortant viruses. After inoculation, no adult hares or chipmunks developed signs of illness. Another reassortant, genotype LAC/LAC/SSH, has exhibited a marked inability to infect chipmunks and was avirulent for mice, probably because of a mutation in its L RNA segment. In chipmunks inoculated with reassortant viruses, neither the durations of viremias nor the maximum titers attained were demonstrably different from those found after inoculation with the parent LAC or SSH viruses. However, viruses with a SSH M RNA induced a higher viremia in snowshoe hares than viruses with a LAC M RNA. Also, the amount of virus needed to produce a detectable infection of chipmunks or hares was lower for viruses in which the M RNA came from the SSH parent than for those viruses having a LAC M RNA. Convalescent phase chipmunk and hare sera from animals infected with SSH virus or reassortant viruses having a SSH M RNA, neutralized parental SSH virus more effectively than did LAC virus. Conversely, the sera of animals infected with LAC virus or reassortants having a LAC M RNA, neutralized LAC virus more efficiently than did SSH virus.

Natural La Crosse virus infection in the red fox (Vulpes fulva), gray fox (urocyon cinereoargenteus), raccoon (Procyon lotor), and opossum (Didelphis virginiana).

Natural infection of sentinel red foxes (Vulpes fulva) and free-ranging red foxes, gray foxes (Urocyon cinereoargenteus) and raccoons (Procyon lotor) with La Crosse (LAC) virus was demonstrated. One isolate of LAC virus was obtained from a sentinel red fox in an enzootic area. The viremia titer of the LAC virus-infected red fox was above the threshold of infection for Aedes triseriatus mosquitoes. Antibody responses were measured by the microneutralization test employing four California group viruses: LAC, snowshoe hare, trivittatus, and Jamestown Canyon. Four of six sentinel red foxes developed LAC virus neutralizing antibody. Antibody titers peaked between days 7 and 34 and were still detectable by dat 345 post-infection. Homologous LAC virus antibody titers were consistently 4-fold or greater than heterologous titers. Serological evidence for natural LAC virus infection was demonstrated in 33 of 57 (58%) free-ranging red foxes, 18 of 32 (57%) gray foxes, and 4 of 16 (25%) raccoons. Antibody titers were comparable in free-ranging foxes infected naturally and those infected experimentally by mosquito bite. The prevalence of infection was significantly different (P less than 0.001) for foxes trapped on three distinct areas within the enzootic region. Rates of infection in foxes by area coincided with LAC virus antibody prevalence observed in free-ranging chipmunks and with reported cases of human LAC encephalitis in Wisconsin.

La Crosse viremias in white-tailed deer and chipmunks exposed by injection or mosquito bite.

To further understand the role of wild mammals in the maintenance of La Crosse virus (LACV) in nature, we investigated the effects of inoculation method and virus source on the duration and amplitude of LACV viremia in vertebrate hosts. Earlier work suggested that deer are not sufficiently susceptible to LACV to play an important role in its maintenance. We re-evaluated the susceptibility of deer since subsequent studies showed that they constitute 65% of Aedes triseriatus blood meals, and thus would be exposed frequently to the virus. In our study, deer developed higher and longer viremia following exposure to LACV by infected Ae. triseriatus than those previously reported by inoculation with needle and syringe. However, susceptible Ae. triseriatus that fed on these viremic animals did not become infected. Because a large number of uninfected mosquitoes can feed upon a viremic deer in nature, we believe that deer should not be disregarded completely as a possible amplifier in the LACV transmission cycle. We also infected chipmunks to determine if there were significant differences in viremia response from mosquito delivery of virus to the chipmunk host, compared with artificial exposure by injection. Chipmunks exposed to infected mosquitoes had higher and longer viremias than the ones produced by intramuscular injection of an LACV suspension. These findings show the importance of using LACV infected mosquitoes for transmission experiments in mammals.

Vector and host relationships of California serogroup viruses in western Siberia.

During 1990 and 1991, adult mosquitoes were collected along the Ob River and its tributaries in western Siberia from approximately 51 degrees 18'N to 66 degrees 4'N. Fifteen virus strains were isolated from 74,196 mosquitoes tested in 1,874 pools. These included Tahyna virus from Aedes cataphylla-punctor subgroup (one) and Ae. excrucians (one), and Inkoo (INK) virus from Ae. communis (one), Ae. communis subgroup (one), Ae. hexodontus (two), Ae. punctor subgroup (two), Ae. punctor complex (one), and unidentified Aedes species (three). In addition, a single Ae. euedes yielded a strain of snowshoe hare (SSH) virus and a strain of Getah, an alphavirus. A Bunyamwera serogroup virus was isolated from Ae. excrucians. With the exception of the two isolates from a single mosquito, minimum infection rates among mosquito taxa ranged from 0.4 to 16.7 per 1,000. The INK virus isolates were widely distributed geographically; however, seven of the 10 isolates were from two sites north of the Arctic Circle. During 1991, sera from two mouse species, five vole species, and four shrew species were collected along the upper Ob River for serologic tests. The prevalence of neutralizing antibody to SSH virus in these sera was 80%. Prevalence rates in the four most abundant species were Apodemus agrarius, 73%; Clethrionomys rutilus, 71%; Microtus arvalis, 80%; and Sorex araneus, 91%. This is the first attempt to clarify the vector and vertebrate host relationships of California serogroup viruses in western Siberia.

Distribution of neutralizing antibodies to California and Bunyamwera serogroup viruses in horses and rodents in California.

Neutralization tests were done on sera from 141 horses from high elevation regions of California. Antibody prevalences to Jamestown Canyon, snowshoe hare, and California encephalitis viruses in the California serogroup and Northway virus in the Bunyamwera serogroup were 55%, 43%, 18%, and 46%, respectively. In 51 horses from rural low elevation regions, seroprevalences were 31%, 35%, 35%, and 37%, respectively. Twenty-four horses from a suburban lowland area were seronegative, except for a single horse with a low titer to snowshoe hare virus. Seroprevalence to Jamestown Canyon and snowshoe hare viruses was associated with increasing age. Only 2 of 177 rodents from the Sierra Nevada had antibodies to Northway virus; none had antibodies to Jamestown Canyon or snowshoe hare viruses.

Ecology of Jamestown Canyon virus (Bunyaviridae: California serogroup) in coastal California.

This paper reports the first isolation of Jamestown Canyon (JC) virus from coastal California and the results of tests for antibody to JC virus in mammals living in coastal California. The virus isolation was made from a pool of 50 Aedes dorsalis females collected as adults from Morro Bay, San Luis Obispo County, California. The virus isolate was identified by two-way plaque reduction-serum dilution neutralization tests done in Vero cell cultures. Sera from the mammals were tested for antibody to JC virus by a plaque-reduction serum dilution neutralization method. A high prevalence of JC virus-specific antibody was found in horses and cattle sampled from Morro Bay. This finding is additional evidence for the presence of a virus antigenically identical or closely related to JC virus in Morro Bay and indicates that the vectors of the virus in Morro Bay feed on large mammals. A high prevalence of virus-specific antibody was also found in horses sampled from Marin and San Diego counties. This finding suggests that viruses antigenically identical or closely related to JC virus are geographically widespread in coastal California.

California serogroup virus infections in Wisconsin domestic animals.

A serologic survey and experimental virus transmission studies were done to assess the role of domestic animals as amplifier hosts of La Crosse (LACV) and Jamestown Canyon (JCV) viruses. Serum from 319 cows, 88 dogs, 122 equines, 47 swine, 10 goats, and 4 cats were tested for neutralizing antibody to LACV, JCV, trivittatus (TVTV), and snowshoe hare (SSHV) viruses. Antibody prevalences of LACV, TVTV, and SSHV were less than 10% in all species. Antibody to JCV was detected in all species except cats. Prevalence ranged from 10% in goats and swine to 29% in dogs. No age-associated trends in JCV prevalence were noted. Two of 6 adult dogs, and 2 of 4 pigs inoculated with 6.3-6.5 log10 suckling mouse intracerebral 50% lethal doses (SMICLD50) of LACV developed viremias ranging of less than 1.0-2.9 log10 SMICLD50/ml 1-3 days after inoculation. Of 4 puppies inoculated with LACV, 3 developed fatal infections. Viremias were not detected in 4 cows, 4 ponies, 7 cats, or 6 sheep. Two cats fed LACV infected suckling mice shed virus from the oropharynx for 1 day each. All animals except 1 cow, 1 cat, and 1 sheep had greater than or equal to 4-fold rise in antibody titers. Five additional dogs fed upon by LACV-infected Aedes triseriatus mosquitoes did not develop viremias or antibody and uninfected Ae. triseriatus engorging on the dogs 1-5 days after feeding by infected mosquitoes failed to become infected. Five ponies, 6 calves, 2 ewes, 6 dogs, and 5 piglets were inoculated with 3.6-7.3 log10 SMICLD50 of JCV. None developed detectable viremias, although greater than or equal to 4-fold rises in antibody titers developed in 60% of the ponies, 17% of the calves, 50% of the dogs, and 1 of 2 ewes. None of the pigs developed corresponding rises in antibody titers. We conclude that juvenile and adult animals of the species tested are not efficient amplifier hosts of LACV or JCV, but may be useful sentinels of local virus transmission.

Ovine fetal malformations induced by in utero inoculation with Main Drain, San Angelo, and LaCrosse viruses.

The teratogenic potential of three bunyaviruses, two California serogroup bunyaviruses, LaCrosse virus and San Angelo virus, and a Bunyamwera serogroup member, Main Drain virus, in sheep was studied following in utero inoculation of ewes in early gestation. Although Main Drain virus appeared to be most teratogenic, all three viruses induced a range of lesions including arthrogryposis, hydrocephalus, fetal death, axial skeletal deviations, anasarca, and oligohydramnios. The teratogenic effects of these viruses are identical to those described in ovine infections by Cache Valley and Akabane viruses. Demonstration of a common bunyaviral tropism for fetal tissue infection that results in congenital brain and musculoskeletal malformations provides evidence that human in utero infection by bunyaviruses could result in similar malformations in human infants.

Experimental infection of foxes with Tahyna virus.

Three foxes about 18 weeks old were each subcutaneously inoculated with approximately 200 LD50 dose of extraneurally passaged strain of the Tahyna virus "236". Viremia was demonstrated in all animals during the 48-96 hours interval after infection. The maximum value was 1.31 dex LD50/0.02 ml. Three weeks after infection a seroconversion was revealed by means of hemagglutination-inhibition test (from titre less than 1 : 10 to 1 : 320) and plaque-reduction neutralization test (from titre less than 1 : 4 to titre 1 : 4096 or 1 : 8192).

Experimental infection of the squirrel (Sciurus vulgaris) and the muskrat (Ondatra zibethica) with Tahyna virus (California group, Bunyaviridae).

Two squirrels aged 16 weeks and three muskrats aged 24 weeks were subcutaneously infected with a dose of 400 SMicLD50 of the extraneurally passaged "236" strain of Tahyna virus. Viremia was detected in one squirrel (48 and 96 hours post infection) and in two muskrats (24 and 48 hours p.i.). Seroconversion was demonstrated by plaque-reduction neutralization test (PRNT) 21 days p.i. in all animals.

Seroconversion rates to Jamestown Canyon virus among six populations of white-tailed deer (Odocoileus virginianus) in Indiana.

The annual seroconversion of fawns, yearlings, and adult white-tailed deer (Odocoileus virginianus) to Jamestown Canyon virus (California group) was followed at six Indiana sites from 1981 through 1984. In all, sera from 1,642 deer (515 fawns, 618 yearlings, and 509 adults) were tested for neutralizing antibody to three California serogroup viruses: Jamestown Canyon, La Crosse, and trivittatus. Virtually all deer with specific neutralizing antibody showed evidence of a prior infection with Jamestown Canyon virus; only three deer showed evidence of a prior infection with only La Crosse virus and none showed evidence of an infection with only trivittatus virus. While there were no significant differences in antibody prevalence to Jamestown Canyon virus between yearling and adult deer at any site, fawns had significantly lower antibody prevalences than either of the two older age groups. Significant differences in antibody prevalence were found between northern versus southern populations of white-tailed deer in Indiana, however, no significant differences were found among the four northern populations or between the two southern populations. The mean antibody prevalences in the two southern fawn, yearling, and adult populations were 15%, 38%, and 41% respectively, while the prevalences in the four northern fawn, yearling, and adult populations were 5%, 67%, and 67% respectively. These different prevalences (northern vs. southern) correlate with the higher Jamestown Canyon virus antibody prevalence in human residents of northern Indiana (2-15%) compared to residents of southern Indiana (less than 2%) found in other studies. The significantly lower prevalence of antibody to Jamestown Canyon virus in fawns is attributed to maternal antibody protecting them from a primary infection their first summer. Yearling deer showed high rates of seroconversion following their second summer of life. These results suggest that infection of white-tailed deer in Indiana with Jamestown Canyon virus is a common phenomenon.

Serological evidence of California serogroup virus activity in Oregon.

We wished to demonstrate evidence of the presence of California serogroup viruses in Oregon and to test for the presence of certain other arboviruses in large ungulates. Blood samples from black-tailed deer (Odocoileus hemionus columbianus), mule deer (O. hemionus hemionus), and Roosevelt elk (Cervus elaphus roosevelti) from nine counties in Oregon were tested by serum-dilution plaque reduction neutralization for antibody to California serogroup viruses, including snowshoe hare, California encephalitis, and Jamestown Canyon, as well as to Cache Valley (Bunyamwera serogroup) and Klamath, an ungrouped rhabdovirus. Of 132 samples tested, 60 (46%) were found to be seropositive at a dilution of greater than or equal to 1:10 for at least one of the five different arboviruses. Forty (30%) samples contained antibody to more than one arbovirus, and 15 samples (11%) contained antibody to all five. Of these 15, 14 were from 75 black-tailed deer sera collected in Lincoln County, Oregon. Seropositivity rates for black-tailed deer ranged from 23% to 35%, with all five arboviruses represented. Positive reactions for all five arboviruses were represented among mule deer sera at rates from 5% to 29%. Elk sera were found to be positive for four of the viruses (none for Klamath virus). Although Cache Valley and Klamath viruses have been reported from Oregon, these data represent the first evidence of a California serogroup virus in the state.

Infection of white-tailed deer (Odocoileus virginianus) in Michigan with Jamestown Canyon virus (California serogroup) and the importance of maternal antibody in viral maintenance.

Sera collected from a captive population of white-tailed deer (Odocoileus virginianus) penned in the lower peninsula of Michigan were assayed over a 29-mo period for neutralizing antibody to California serogroup viruses. In all, 130 individual white-tailed deer were bled one to 22 times between June 1983 and November 1985. Of the 130 sampled after active transmission had ceased, or passage of maternal antibody in colostrum had occurred, only one (0.8%), a newborn fawn, had no serum neutralizing antibody to California group viruses. All 18 1-yr-old does sampled acquired specific neutralizing antibody to Jamestown Canyon (JC) virus within a 6-wk period in 1984 and within a 10-wk period in 1985 indicating the prevalence of infection in this nonimmune age group was 100% for 2 successive yr. All 32 2- to 7-yr-old adult does and eight bucks sampled between June 1983 and June 1985 had specific neutralizing antibody to JC virus. No white-tailed deer had specific neutralizing antibody to trivittatus or La Crosse/snowshoe hare viruses at this study site. In 1984 and 1985, 78% and 63% of the adult does respectively exhibited significant anamnestic responses; all 19 adult does sampled over two seasons (between October 1983 and June 1985) showed a significant anamnestic response during at least 1 of the 2 yr. One-third of adult does with significant springtime antibody titer increases apparently experienced reexposure prior to the emergence of aedine mosquitoes, suggesting an alternate vector may overwinter at this site and transmit viruses in early spring. Specific neutralizing antibody was detected in 98% (66/67) of nursing fawns bled within 5 wk of birth in May-June 1984 and 1985, including three of three nursing fawns bled within 24-96 hr of birth. Of the 66 newborn fawns with specific neutralizing antibody to JC virus in June 1984 and 1985, 95% (54/57) of the surviving fawns lost maternal antibody and had no measurable titer when sampled 20-24 wk after birth, however. Serum antibody titers in 25 newborn (1984-cohort) fawns and their mothers and titers in 38 newborn (1985-cohort) fawns and their mothers were significantly correlated at the 5% and 1% levels respectively, suggesting that maternal antibody rather than a naturally acquired infection was the source of immunity in these suckling fawns.(ABSTRACT TRUNCATED AT 400 WORDS)

Serologic evidence of Jamestown Canyon virus infection in white-tailed deer populations from Connecticut.

We determined the prevalence and distribution of Jamestown Canyon (JC) virus antibody in white-tailed deer (Odocoileus virginianus) populations in Connecticut, USA. Sera were collected from hunter-killed deer during 1993. Antibody to JC virus was detected by enzyme-linked immunosorbent assay (ELISA) in 92 (21%) of 446 deer sera, and was uniformly distributed among geographic sites. Twenty-one ELISA-positive sera were tested and confirmed positive by plaque reduction neutralization testing. This represents the first serologic evidence of JC virus in a reservoir host population from the northeastern United States. No cross-reactivity was seen with California encephalitis, Keystone, or snowshoe hare viruses, but a varying degree of cross-reactivity was obtained with Guaroa, Jerry Slough, La-Crosse, San Angelo, and trivittatus viruses. We conclude from this investigation and previous isolations of JC virus from mosquitoes in the state that JC virus occurs enzootically in Connecticut.

Serological evidence of California group and Cache Valley virus infection in Minnesota white-tailed deer.

Blood samples were obtained from 138 white-tailed deer (Odocoileus virginianus) harvested at three sites surrounding the greater Minneapolis-St. Paul, Minnesota, metropolitan area (USA) and tested for neutralizing antibody to Cache Valley virus and three California serogroup (Jamestown Canyon, La Crosse, trivittatus) viruses (Bunyaviridae). Deer at each site had neutralizing antibody to one or more California serogroup viruses and/or Cache Valley virus. The majority of adult deer (85%) had antibody to both a California serogroup virus and Cache Valley virus. Antibody prevalence varied significantly with age of the deer. Fawns had a significantly lower prevalence of antibody to either a California serogroup (17%) or Cache Valley virus (39%) than did older (greater than 1-yr-old) deer (89% for a California serogroup virus and 91% for Cache Valley virus). The geometric mean titers of antibody in fawns to California serogroup (1:6) and Cache Valley viruses (1:17) were also less than that seen in older animals (1:11 and 1:28 for California serogroup and Cache Valley viruses, respectively). Of 76 older deer with antibody to the California serogroup, 91% had antibody specific for Jamestown Canyon virus. Jamestown Canyon is the primary California serogroup virus circulating in the suburban/rural Minneapolis-St. Paul area. Transmission occurs in an enzootic pattern similar to that documented in Indiana and Michigan. Cache Valley virus also appears to be enzootically transmitted in this area. However, the impact on domestic or wild animal populations is unknown.