California encephalitis virus activity in mosquitoes and horses in southern Ontario, 1975.

A study was undertaken in 1975 to determine California encephalitis virus activity in southern Ontario. Three thousand and sixty-one mosquitoes, primarily Aedes species, were divided into 104 pools and inoculated into suckling mice. Isolates of snowshoe hare virus were obtained from one pool each of Aedes fitchii and A. triseriatus mosquitoes collected in the Guelph area. Serological testing of horse sera revealed extensive virus activity in southern Ontario and indicated that horses may serve as excellent monitors for California encephalitis virus.     

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.     

Antibodies to arboviruses in an Alaskan population at occupational risk of infection

A total of 435 United States Geological Survey and United States Forest Service workers in Alaska were studied for serologic evidence of past infections with four arboviruses known or suspected to be human pathogens. Of the personnel tested, 36 (8.3%) had the neutralizing antibody to Jamestown Canyon but not snowshoe hare virus, 6 (1.4%) had the antibody to snowshoe hare but not Jamestown Canyon virus, 53 (12.2%) had the antibody to both viruses, 17 (3.9%) had the antibody to Northway virus, and 15 (3.4%) had the antibody to Klamath virus. The indices most significantly correlated with presence of the Jamestown Canyon and snowshoe hare antibodies were the amount of fieldwork (p less than 0.001 for both antibodies) and the duration of employment by the agencies (p less than 0.0001 for Jamestown Canyon and 0.004 for snowshoe hare). The antibody to the four arboviruses also correlated strongly with a history of travel in certain remote or wilderness areas in Alaska (p values ranged from less than 0.001 to 0.086).     

Serum antibody prevalence for Herpesvirus sylvilagus, Bacillus piliformis and California serogroup arboviruses in cottontail rabbits from Pennsylvania

A serologic survey of 60 eastern cottontail rabbits (Sylvilagus floridanus) from three counties in Pennsylvania was conducted in March 1983. Serum antibody prevalences for Herpesvirus sylvilagus and La Crosse virus (California serogroup) were less than 4%. There was no evidence of previous exposure to either Jamestown Canyon or snowshoe hare viruses (California serogroup). Antibody to trivittatus virus (California serogroup) was found in 60% of the 20 cottontails from York County. No cottontails had antibodies to Bacillus piliformis, the etiologic agent of Tyzzer's disease.     

Serologic survey for selected microbial agents in mammals from Alberta, 1976

Blood samples were taken from humans and several species of free-ranging wild mammals from five different geographic areas of Alberta, Canada. Sera were tested for antibody to eastern equine encephalitis (EEE) virus, western equine encephalitis (WEE) virus, St. Louis encephalitis (SLE) virus, Powassan (POW) virus, the snowshoe hare (SSH) strain of the California group (CAL) of viruses, Northway (NOR) virus, Klamath (KLA) virus, infectious bovine rhinotracheitis (IBR) virus, and two bacteria, Brucella abortus and Francisella tularensis. CAL antibody was found in 63% of 11 snowshoe hares (Lepus americanus), 33% of 167 black bears (Ursus americanus), and 19% of 55 humans (Homo sapiens). NOR antibody was found in 0.4% of 258 hares, 11% of 9 bighorn sheep (Ovis canadensis), 20% of 44 moose (Alces alces), 4% of 56 bears, 14% of 22 woodland caribou (Rangifer tarandus), and 2% of 50 humans. IBR antibody was detected in 14% of 14 moose. B. abortus antibody was found in 1% of 283 bears. F. tularensis antibody was detected in 2% of 52 humans. These findings represent extension of: (1) the natural host range of IBR, CAL, and NOR; (2) the geographical distribution of NOR infection in North America; and (3) the geographical distribution of CAL infection within Alberta.     

Activity of California encephalitis group viruses in Entrelacs (province of Quebec, Canada)

During the summer of 1979, indicator rabbits were placed in three sites in Entrelacs (Laurentian area, province of Quebec) and mosquitoes were collected in order to monitor arbovirus activity in the area. Eight seroconversions to California encephalitis (CE) group viruses were detected in rabbits during June, July, and August. Twenty-five strains identified as members of the CE group were isolated: 3 were obtained from viremic rabbit sera, 1 from adult Aedes communis reared in the laboratory from field-collected larvae, and 21 from mosquito pools. Twenty-two of these were typed as snowshoe hare (SSH) virus. No evidence of La Crosse (LAC) virus was detected but three strains belonging to the CE group showed antigenic properties different from reference SSH, LAC, or Jamestown Canyon (JC) viruses. One isolate identified as Flanders virus was obtained from Culex pipiens. Three mosquito species (A. communis, A. punctor, and A. excrucians) were involved in the transmission cycle of SSH virus in Entrelacs. This is the first report, in the province of Quebec, of SSH isolation from animal sera and the first demonstration of its transovarial transmission.     

Virus isolations from mosquitoes in southern Ontario, 1976 and 1977

Following the 1975 epidemic of St. Louis encephalitis (SLE) in Ontario, programs were instituted to monitor virus activity in mosquito populations during 1976 and 1977. Mosquitoes were trapped with CDC light traps and CO2 cone traps, pooled by species, and tested for virus by intracerebral inoculation of suckling mice. In 1976, 51 175 mosquitoes were tested. SLE virus was isolated from two mixed pools of Culex pipiens--C. restuans mosquitoes. Five isolations of California serogroup viruses were made. Three of these were trivittatus virus, which has not been demonstrated previously in Canada, and the other two were snowshoe have virus. Other viruses isolated in 1976 were a virus antigenically identical to the virus of infectious bursal disease of chickens and 34 Flanders viruses. In 1977, 34 428 mosquitoes were tested. Flanders virus was isolated most frequently, from pools of mixed C. pipiens--C. restuans mosquitoes. The only other isolate was a Bunyamwera group virus, Cache Valley virus. This virus has not been reported previously in Ontario.     

California encephalitis virus development in mosquitoes as revealed by transmission studies, immunoperoxidase staining, and electron microscopy.

Isolates of the snowshoe hare subtype of California encephalitis (CE) virus from Yukon mosquitoes during 1972 and 1973 were transmitted by bites of Aedes aegypti mosquitoes after 4 to 5 weeks of extrinsic incubation at 55 degrees F after intrathoracic injection, and the 1973 strain was transmitted after mosquitoes were fed virus and held for 3 to 4 weeks at 75 degrees F. Antigen of a 1971 isolate of CE virus (Marsh Lake 23) was detected in salivary glands of infected mosquitoes by the immunoperoxidase technique, using highly purified antiserum before and after conjugation with horseradish peroxidase, plus the use of orthotolidine as a substitute for benzidine. enveloped virions 45 nm in diameter were observed in thin sections of salivary glands of Culiseta inornata mosquitoes 59 days after intrathoracic injection with the 1971 isolate, afterincubation at 55 degrees F.     

Nucleotide Sequence Analyses and Predicted Coding of Bunyavirus Genome RNA Species

We performed 3′ RNA sequence analyses of [³²P]pCp-end-labeled La Crosse (LAC) virus, alternate LAC virus isolate L74, and snowshoe hare bunyavirus large (L), medium (M), and small (S) negative-stranded viral RNA species to determine the coding capabilities of these species. These analyses were confirmed by dideoxy primer extension studies in which we used a synthetic oligodeoxynucleotide primer complementary to the conserved 3′-terminal decanucleotide of the three viral RNA species (Clerx-van Haaster and Bishop, Virology 105:564-574, 1980). The deduced sequences predicted translation of two S-RNA gene products that were read in overlapping reading frames. So far, only single contiguous open reading frames have been identified for the viral M- and L-RNA species. For the negative-stranded M-RNA species of all three viruses, the single reading frame developed from the first 3′-proximal UAC triplet. Likewise, for the L-RNA of the alternate LAC isolate, a single open reading frame developed from the first 3′-proximal UAC triplet. The corresponding L-RNA sequences of prototype LAC and snowshoe hare viruses initiated open reading frames; however, for both viral L-RNA species there was a preceding 3′-proximal UAC triplet in another reading frame that was followed shortly afterward by a termination codon. A comparison of the sequence data obtained for snowshoe hare virus, LAC virus, and the alternate LAC virus isolate showed that the identified nucleotide substitutions were sufficient to account for some of the fingerprint differences in the L-, M-, and S-RNA species of the three viruses. Unlike the distribution of the L- and M-RNA substitutions, significantly fewer nucleotide substitutions occurred after the initial UAC triplet of the S-RNA species than before this triplet, implying that the overlapping genes of the S RNA provided a constraint against evolution by point mutation. The comparative sequence analyses predicted amino acid differences among the corresponding L-, M-, and S-RNA gene products of snowshoe hare virus and the two LAC virus isolates.     

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.     

Serologic evidence of arbovirus infections in humans and wild animals in Alaska

Blood samples were collected from humans and several species of free-ranging wild animals in Alaska. Sera were tested for antibody to Jamestown Canyon (JC), snowshoe hare (SSH), Northway (NOR), Klamath (KLA), Sakhalin (SAK), Great Island (GI), and Silverwater (SIL) virus. JC antibody was found in 54% of 121 human, 89% of 97 bison (Bison bison), 51% of 84 Dall sheep (Ovis dalli), 43% of 68 snowshoe hare (Lepus americanus), and 3% of 33 arctic fox (Alopex lagopus) sera. SSH antibody was found in 42% of 121 human, 89% of 97 bison, 41% of 84 Dall sheep, and 65% of 68 snowshoe hare sera. NOR antibody was found in 14% of 121 human, 94% of 97 bison, 84% of 84 Dall sheep, 43% of 69 caribou (Rangifer tarandus), 3% of 68 snowshoe hare, 48% of 64 grizzly bear (Ursus arctos), 3% of 33 arctic fox, and 78% of 27 moose (Alces alces) sera. KLA antibody was found in 5% of 121 human and 40% of 97 bison sera. SAK antibody was found in 2% of 97 bison and 3% of 33 arctic fox sera. GI antibody was found in 1% of 97 bison sera. No SIL antibody was found in any sera tested. Thus the natural host ranges of JC, SSH, NOR, and KLA viruses have been extended by inference from the occurrence of antibody.     

Relationship Between Fecal Pellet Counts and Snowshoe Hare Density in Western Wyoming

ABSTRACT Snowshoe hares (Lepus americanus) are an important prey species for Canada lynx (Lynx canadensis) and are considered critical for lynx population persistence. Determination of snowshoe hare distribution and abundance is needed by land management agencies for lynx conservation. An accepted approach for estimating snowshoe hare abundance is the use of fecal-pellet plot counts. Locally derived regression equations are preferred for accurate calibration of pellet counts to snowshoe hare density due to local differences in pellet deposition and decomposition. We used linear regression to examine correlations between snowshoe hare density, as determined by mark–recapture estimates, and pellet plot counts on both uncleared plots and annually cleared plots on the Bridger-Teton National Forest, western Wyoming, USA. We found significant correlations between snowshoe hare density estimates and fecal pellet counts for both uncleared and annually cleared pellet counts; however, the relationship was stronger (higher r) when using pellet counts from annually cleared plots. In addition, we found that adjusting the buffer size by omitting hard habitat edges (not used by hares) around trapping grids improved correlations between snowshoe hare density and fecal pellet counts for both uncleared plots and annually cleared plots. Though precision is sacrificed when using uncleared plots, they may be useful as a coarse index of habitat use by snowshoe hares. Our derived regression equations may be useful to identify important foraging habitat for Canada lynx in western Wyoming. Land managers responsible for conserving snowshoe hare habitat in western Wyoming may use these equations to monitor changes in hare populations among habitats and during prescribed management actions.     

Multiscale habitat relationships of snowshoe hares (Lepus americanus) in the mixed conifer landscape of the Northern Rockies,USA: Cross‐scale effects of horizontal cover with implications for forest management

Snowshoe hares (Lepus americanus) are an ecologically important herbivore because they modify vegetation through browsing and serve as a prey resource for multiple predators. We implemented a multiscale approach to characterize habitat relationships for snowshoe hares across the mixed conifer landscape of the northern Rocky Mountains, USA. Our objectives were to (1) assess the relationship between horizontal cover and snowshoe hares, (2) estimate how forest metrics vary across the gradient of snowshoe hare use and horizontal cover, and (3) model and map snowshoe hare occupancy and intensity of use. Results indicated that both occupancy and intensity of use by snowshoe hares increased with horizontal cover and that the effect became stronger as intensity of use increased. This underscores the importance of dense horizontal cover to achieve high use, and likely density, of snowshoe hares. Forest structure in areas with high snowshoe hare use and horizontal cover was characterized as multistoried with dense canopy cover and medium‐sized trees (e.g., 12.7–24.4 cm). The abundance of lodgepole pine (Pinus contorta) was associated with snowshoe hare use within a mixed conifer context, and the only species to increase in abundance with horizontal cover was Engelmann spruce (Picea engelmannii) and subalpine fir (Abies lasiocarpa). Our landscape‐level modeling produced similar patterns in that we observed a positive effect of lodgepole pine and horizontal cover on both occupancy and use by snowshoe hares, but we also observed a positive yet parabolic effect of snow depth on snowshoe hare occupancy. This work is among the first to characterize the multiscale habitat relationships of snowshoe hares across a mixed conifer landscape as well as to map their occupancy and intensity of use. Moreover, our results provide stand‐ and landscape‐level insights that directly relate to management agencies, which aids in conservation efforts of snowshoe hares and their associated predators.     

UV-B-induced plant stress as a possible cause of ten-year hare cycles

Predation has been assumed to be a necessary factor in the ten-year population cycle of the snowshoe hare (Lepus americanus) and Canadian lynx (Lynx canadensis). The UV-B-induced plant stress hypothesis, in contrast, predicts that hare performance, especially reproduction, is negatively related to sunspot numbers, because production of UV-B-protective phenolics in food plants in periods of low sunspot activity, when the ozone layer is thin, increases the availability of amino acids and reduces the amount of phenolics that protect against herbivores. In accordance with the UV-B-induced plant stress hypothesis, and despite the absence of predators that have been assumed to be necessary for hare cycles, mountain hare (Lepus timidus) populations in Norway fluctuate in close synchrony with snowshoe hare populations in Alberta and the Yukon, Canada. When adjusting for the phase of the hare cycle, the natality of snowshoe hare in Alberta 1962–1976 was negatively related to sunspot numbers with a time lag of two years. It is concluded that delayed responses to UV-B-induced changes in plant chemistry during the sunspot cycle is a possible cause of ten-year cycles of hares and other herbivores, for example grouse and forest moths.     

A plant toxin mediated mechanism for the lag in snowshoe hare population recovery following cyclic declines

A necessary condition for a snowshoe hare population to cycle is reduced reproduction after the population declines. But the cause of a cyclic snowshoe hare population's reduced reproduction during the low phase of the cycle, when predator density collapses, is not completely understood. We propose that moderate‐severe browsing by snowshoe hares upon preferred winter‐foods could increase the toxicity of some of the hare's best winter‐foods during the following hare low, with the result being a decline in hare nutrition that could reduce hare reproduction. We used a combination of modeling and experiments to explore this hypothesis. Using the shrub birch Betula glandulosa as the plant of interest, the model predicted that browsing by hares during a hare cycle peak, by increasing the toxicity B. glandulosa twigs during the following hare low, could cause a hare population to cycle. The model's assumptions were verified with assays of dammarane triterpenes in segments of B. glandulosa twigs and captive hare feeding experiments conducted in Alaska during February and March 1986. The model's predictions were tested with estimates of hare density and measurements of B. glandulosa twig growth made at Kluane, Yukon from 1988–2008. The empirical tests supported the model's predictions. Thus, we have concluded that a browsing‐caused increase in twig toxicity that occurs during the hare cycle's low phase could reduce hare reproduction during the low phase of the hare cycle.     

Snowshoe hare demography during a cyclic population low

1. Snowshoe hare ( Lepus americanus Erxleben) populations were studied in south-west Yukon during the low phase of the 10-year population cycle. Food availability and predator abundance were manipulated in a factorial design to determine the importance of each factor in hare dynamics during this phase.
2. Food was abundant during the low phase, and snowshoe hares were not food limited.
3. Survival of hares was higher than at any other phase of the cycle, and predators were scarce, but >75% of hare deaths resulted from predation.
4. Food addition resulted in higher hare densities and better body condition than on control sites. There were no observable effects of food addition on population rate of increase, recruitment, survival or age structure.
5. Mammalian predator reduction resulted in higher hare densities, higher survival, better body condition and an older age structure. Relative to control populations, recruitment was lower and population rates of increase similar.
6. The joint manipulation of food addition + predator reduction had greater positive effects on hare density and body condition than either single factor manipulation. Survival was better than on control sites, and the age structure was older than on control sites. Population rates of increase were similar, but recruitment was higher on the control areas.
7. We conclude that snowshoe hare dynamics at the low of the cycle are dominated by the interaction of food and predation. Risk of predation also had indirect effects on snowshoe hare age structure and body condition.     

Regulation of boreal forest understory vegetation: the roles of resources and herbivores

To understand inter-trophic linkages between components of the boreal forest understory vegetation, three hypotheses were tested: survival, growth and abundance of grasses and legumes were controlled by (i) resource availability alone, (ii) by herbivores alone, and (iii) by both resource availability and herbivores. The hypotheses were tested using three experimental treatments – fertilization, herbivore exclusion, and fertilization plus herbivore exclusion – in three areas having different densities of resident herbivores, mostly snowshoe hares and ground squirrels. The highest density of snowshoe hares is comparable to natural levels during peaks in the snowshoe hare cycle. As the density of herbivores increased so too did the level of response by the measured variables – survival, growth of transplants and leaf area index of established vegetation. In general, fertilization resulted in a decrease in survival and growth of transplants, and fences increased survival and growth; both responses were more noticeable at higher herbivore densities. Fertilizer and herbivore exclosure fences had only negligible effects on established grass and legume abundance at all hare densities. We have shown that some hypotheses of vegetation regulation are over-simplified because different species groups (i.e., grasses and legumes) are regulated by different factors, at different life history stages, and sometimes these factors act in opposing directions. We argue that during the increase phase and peak of the snowshoe hare cycle (high herbivore density), growth and survival of establishing plants is regulated by herbivores. During the decline and low phases of the snowshoe hare cycle herbivores will have little impact on early life stages, whereas the established, mature, vegetation will be resource-regulated. Because of the variability in responses to the same manipulations we may begin to understand which plant life history stages are most vulnerable to consumer and resource regulation, the magnitudes of these sources of regulation at each of these stages, and how these vary among species groups and types of environments.     

Three unique viral RNA species of snowshoe hare and La Crosse bunyaviruses.

Two-dimensional gel electrophoreses of RNase T1-derived oligonucleotides of the three individual RNA segments of the bunyavirus snowshow hare virus indicate that its three RNA segments possess distinct nucleotide sequences. The fingerprints of the RNA species of snowshoe hare virus differ from those of the antigenically closely related La Crosse virus. Three viral RNA species have been identified in preparations of Melao and Trivittatus as well as snowshoe hare, Lumbo, and La Crosse bunyaviruses.     

Prevalence of neutralizing antibody to Jamestown Canyon virus (California group) in populations of elk and moose in northern Michigan and Ontario, Canada

Blood samples were collected from free-ranging elk (Cervus elaphus) harvested in Michigan's northern Lower Peninsula, from moose (Alces alces) relocated from Ontario's Algonquin Provincial Park to Michigan's Upper Peninsula, and from moose from Michigan's Isle Royale National Park. Sera were tested by serum dilution neutralization tests in Vero cell culture for neutralizing antibody to California serogroup viruses, in particular Jamestown Canyon (JC), La Crosse/snowshoe hare (LAC/SSH), and trivittatus (TVT) viruses. Specific neutralizing antibody to JC virus was detected in 71% of 31 and 65% of 20 moose from Algonquin and Isle Royale, respectively. An additional six moose from Algonquin and five from Isle Royale showed evidence of multiple infection. One juvenile moose from Isle Royale had specific neutralizing antibody to TVT virus. Specific neutralizing antibody to JC virus was detected also in 54% of 50 elk from Michigan; 20 of the 50 elk showed evidence of multiple infection. While no single serum sample showed specific neutralizing antibody only to LAC/SSH virus, its presence in sera from some animals may have been masked by the high prevalence of antibody to JC virus.     

Novel wildlife in the Arctic: the influence of changing riparian ecosystems and shrub habitat expansion on snowshoe hares

Warming during the 20th century has changed the arctic landscape, including aspects of the hydrology, vegetation, permafrost, and glaciers, but effects on wildlife have been difficult to detect. The primary aim of this study is to examine the physical and biological processes contributing to the expanded riparian habitat and range of snowshoe hares (Lepus americanus) in northern Alaska. We explore linkages between components of the riparian ecosystem in Arctic Alaska since the 1960s, including seasonality of stream flow, air temperature, floodplain shrub habitat, and snowshoe hare distributions. Our analyses show that the peak discharge during spring snowmelt has occurred on average 3.4 days per decade earlier over the last 30 years and has contributed to a longer growing season in floodplain ecosystems. We use empirical correlations between cumulative summer warmth and riparian shrub height to reconstruct annual changes in shrub height from the 1960s to the present. The effects of longer and warmer growing seasons are estimated to have stimulated a 78% increase in the height of riparian shrubs. Earlier spring discharge and the estimated increase in riparian shrub height are consistent with observed riparian shrub expansion in the region. Our browsing measurements show that snowshoe hares require a mean riparian shrub height of at least 1.24–1.36 m, a threshold which our hindcasting indicates was met between 1964 and 1989. This generally coincides with observational evidence we present suggesting that snowshoe hares became established in 1977 or 1978. Warming and expanded shrub habitat is the most plausible reason for recent snowshoe hare establishment in Arctic Alaska. The establishment of snowshoe hares and other shrub herbivores in the Arctic in response to increasing shrub habitat is a contrasting terrestrial counterpart to the decline in marine mammals reliant on decreasing sea ice.