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Mosquito-borne La Crosse virus (LACV; family: Peribunyaviridae) is the leading cause of pediatric arboviral encephalitis in the United States, with clinical cases generally centered in the Midwest and Appalachian regions. Incidence of LACV cases in Appalachian states has increased, such that the region currently represents the majority of reported LACV cases in the USA. The amount of reported LACV cases from Virginia, however, is minimal compared to neighboring states such as North Carolina, West Virginia, and Tennessee, and non-Appalachian regions of Virginia are understudied. Here we examine the hypothesis that LACV is circulating widely in Virginia, despite a low clinical case report rate, and that the virus is circulating in areas not associated with LACV disease. In this study, we screened local mammalian wildlife in northwestern counties of Virginia using passive surveillance via patients submitted to wildlife rehabilitation centers. Blood sera (527 samples; 9 species, 8 genera) collected between October 2019 and December 2022 were screened for neutralizing antibodies against LACV, indicating prior exposure to the virus. We found an overall LACV seroprevalence of 1.90% among all wild mammals examined and reveal evidence of LACV exposure in several wild species not generally associated with LACV, including eastern cottontails and red foxes, along with established reservoirs, eastern gray squirrels, although there was no serological evidence in chipmunks. These data indicate the circulation of LACV in Virginia outside of Appalachian counties, however, at a lower rate than reported for endemic areas within the state and in other states.

Over the past decade, Haemaphysalis longicornis, the Asian longhorned tick, has undergone a geographic range expansion in the United States, from its historical range in east Asia. This tick has been characterized by its frequent parasitism of livestock, an ability to reproduce through parthenogenesis, and its ability to transmit a variety of vector-borne pathogens to livestock, wildlife, and human hosts in its native geographic range. Thus far in the United States, 17 states have reported H. longicornis populations, including 38 counties in Virginia. These numbers come from presence-absence reports provided to the U.S. Department of Agriculture, but little has been reported about this ticks' seasonality in Virginia or its habitat preferences. Our current study detected H. longicornis populations in seven of the nine surveyed counties in Virginia. Haemaphysalis longicornis were observed in multiple habitat types including mixed hardwood forests and pastures, with abundant H. longicornis populations detected at one particular pasture site in Wythe County. This study also attempted to investigate environmental conditions that may be of importance in predicting tick presence likelihood. While sample size limited the scope of these efforts, habitat type and climatic metrics were found to be important indicators of H. longicornis collection success and abundance for both the nymphal and larval life stages. This current study reports useful surveillance data for monitoring these tick populations as they become established in the western half of Virginia and provides insight into their current distribution and maintenance over a large study region.

Characteristic of the Anthropocene, human impacts have resulted in worldwide losses in forested land cover, which can directly and indirectly drive biodiversity loss. The global illicit drug trade is one source of deforestation directly implicated with habitat loss in Central America, typically for drug trafficking and livestock production for money laundering. Given reports of deforestation in Central America linked to narcotraffic, we explored vegetation changes within Guatemala’s highly biodiverse Maya Biosphere Reserve by examining trends suggestive of deforestation in a protected area. As such, we collected satellite-derived data in the form of enhanced vegetation index (EVI), as well as history of burned areas, published human-“footprint” data, official population density, and artificial light activity in Laguna del Tigre National Park from 2002 to 2020 for descriptive analysis. We found consistent reductions in EVI and trends of anomalous losses of vegetation despite a baseline accounting for variation within the park. Analyses revealed weak correlations (R2 ≤ 0.26) between EVI losses and official sources of anthropogenic data, which may be attributable to the data’s limited spatial and temporal resolution. Alarmingly, simple analyses identified vegetation losses within a protected area, thus emphasizing the need for additional monitoring and science-based, but interdisciplinary policies to protect this biodiversity hotspot.

Here we report the first detection and confirmation of Powassan virus (POWV) (family: Flaviridae) in Ixodes scapularis ticks collected from Appalachian Virginia. Ixodes scapularis ticks were collected from vegetation across field sites in eight counties of western Virginia from June 2019 to April 2021. From these collections, one nymph and one adult male I. scapularis were determined to be positive for POWV using real-time RT-PCR and Sanger sequencing. Both positive ticks were collected from Floyd county, VA, at residential sites; the nymph in June 2020 and the adult male in April 2021. The presence of POWV in Virginia in its natural tick vector is crucial knowledge in beginning to understand the movement and transmission of this pathogen into new geographical areas and the risk it poses to medical and veterinary health.

Background. African animal trypanosomiasis (AAT) affects livestock productivity in sub-Saharan Africa. This study aimed to determine cattle AAT’s prevalence and associated risk factors in Lambwe Valley, Kenya. Methods. In a cross-sectional survey, livestock owners were recruited from four villages of Lambwe in Homa Bay, Kenya. Blood samples were collected from the jugular veins of cattle, and buffy coat smears were examined under a microscope. Parasites were further detected using polymerase chain reaction (PCR). Using a semistructured questionnaire, livestock owners were interviewed on their knowledge of AAT and control practices. Chi-square and multilevel models were used for the analysis. Results. The overall prevalence was 15.63% (71/454). Trypanosoma vivax 10.31% and T. congolense Savannah 6.01% were the common species and subspecies. A total of 61 livestock keepers were involved in the study. Of these, 91.80% (56/61) knew AAT, and 90.16% (55/61) could describe the symptoms well and knew tsetse fly bite as transmission mode. Self-treatment (54.09%; 33/61) was common, with up to 50.00% of the farmers using drugs frequently. Isometamidium (72.13%; 44/61) and diminazene (54.09%; 33/61) were drugs frequently used. Although 16.39% (10/61) of the farmers claimed to use chemoprophylactic treatment, 6/10 did not use the right drugs. Animals (92.1%; 58/63) with clinical signs had positive infections. Villages closer to the national park recorded a higher prevalence. Infections were higher in cattle owned by those self-treating (27.23%; 58/213), those using drug treatment without vector control (27.62%; 50/181), those using single-drug therapy, and those practicing communal grazing (20.00%; 59/295). Clinical signs strongly associate with positive infections under multilevel modeling. Conclusion. Cattle trypanosomiasis is prevalent in the Lambwe region of Kenya. This is influenced by inappropriate control practices, communal grazing, and the proximity of farms to the nati

La Crosse virus (LACV) is the leading cause of pediatric viral encephalitis in North America, and is an important public health pathogen. Historically, studies involving LACV pathogenesis have focused on lineage I strains, but no former work has explored the pathogenesis between or within lineages. Given the absence of LACV disease in endemic regions where a robust entomological risk exists, we hypothesize that some LACV strains are attenuated and demonstrate reduced neuroinvasiveness. Herein, we compared four viral strains representing all three lineages to determine differences in neurovirulence or neuroinvasiveness using three murine models. A representative strain from lineage I was shown to be the most lethal, causing >50% mortality in each of the three mouse studies. However, other strains only presented excessive mortality (>50%) within the suckling mouse neurovirulence model. Neurovirulence was comparable among strains, but viruses differed in their neuroinvasive capacities. Our studies also showed that viruses within lineage III vary in pathogenesis with contemporaneous strains, showing reduced neuroinvasiveness compared to an ancestral strain from the same U.S. state (i.e., Connecticut). These findings demonstrate that LACV strains differ markedly in pathogenesis, and that strain selection is important for assessing vaccine and therapeutic efficacies.

Ticks are susceptible to environmental conditions and, to ensure survival during winter conditions, they adopt a wide variety of physiological and behavioral adaptations including utilization of a suitable niche with insulation (e.g., leaf coverage). To investigate the potential overwintering survival of three tick populations emerging within Appalachian Virginia (Haemaphysalis longicornis, Amblyomma americanum, and Amblyomma maculatum), both a laboratory experiment assessing super-cooling points and a two-factor (elevation and insulation coverage) field experiment assessing overwintering survivability were conducted across a natural southwestern Virginian winter (2020–2021). Dermacentor variabilis adults were included in this study as an example of a well-established species in this region known to overwinter in these conditions. Our study indicated that A. americanum and H. longicornis wintering tolerance is based on life stage rather than external factors such as insulation (e.g., leaf litter) and elevation. Amblyomma maculatum was more likely to survive without insulation. The ability to withstand the extreme temperatures of new regions is a key factor determining the survivability of novel tick species and is useful in assessing the invasion potential of arthropod vectors.

Background: Natural infections with soil-transmitted nematodes occur in non-human primates (NHPs) and have the potential to cross primate-species boundaries and cause diseases of significant public health concern. Despite the presence of NHPs in most urban centres in Kenya, comprehensive studies on their gastrointestinal parasites are scant. Objective: Conduct a cross-sectional survey to identify zoonotic nematodes in free-ranging NHPs found within four selected urban and peri-urban centres in Kenya. Methods: A total of 86 NHPs: 41 African green monkeys [AGMs] (Chlorocebus aethiops), 30 olive baboons (Papio anubis), 5 blue monkeys (Cercopithecus mitis stuhlmanni) and 10 red-tailed monkeys (Cercopithecus ascanius) were sampled once in situ and released back to their habitat. Microscopy was used to identify nematodes egg and larvae stages in the samples. Subsequently, PCR coupled with high-resolution melting (PCR-HRM) analysis and sequencing were used to identify nodule worms. Results: NHPs inhabiting densely populated urban environs in Kenya were found infected with a rich diversity of nematodes including three potentially zoonotic nematodes including Oesophagostomum stephanostomum, Oesophagostomum bifurcum and Trichostrongylus colubriformis and co-infections were common. Conclusion: Phylogenetic analysis showed that O. stephanostomum from red-tailed and blue monkeys have a close evolutionary relatedness to human isolates suggesting the zoonotic potential of this parasite. Moreover, we also report the first natural co-infection of O. bifurcum and O. stephanostomum in free-ranging AGMs.

La Crosse virus (LACV) is the leading cause of pediatric viral encephalitis in North America, and is an important public health pathogen. Historically, studies involving LACV pathogenesis have focused on lineage I strains, but no former work has explored the pathogenesis between or within lineages. Given the absence of LACV disease in endemic regions where a robust entomological risk exists, we hypothesize that some LACV strains are attenuated and demonstrate reduced neuroinvasiveness. Herein, we compared four viral strains representing all three lineages to determine differences in neurovirulence or neuroinvasiveness using three murine models. A representative strain from lineage I was shown to be the most lethal, causing >50% mortality in each of the three mouse studies. However, other strains only presented excessive mortality (>50%) within the suckling mouse neurovirulence model. Neurovirulence was comparable among strains, but viruses differed in their neuroinvasive capacities. Our studies also showed that viruses within lineage III vary in pathogenesis with contemporaneous strains, showing reduced neuroinvasiveness compared to an ancestral strain from the same U.S. state (i.e., Connecticut). These findings demonstrate that LACV strains differ markedly in pathogenesis, and that strain selection is important for assessing vaccine and therapeutic efficacies.

The Asian tiger mosquito (Aedes albopictus) is an important vector of a number of arboviruses, including Zika (ZIKV), dengue (DENV), and chikungunya (CHIKV) viruses, and has recently expanded its range in the eastern United States to southern New England and New York. Given the recent establishment and proliferation of Ae. albopictus in this region and the increasing amount of international travel between the United States and endemic countries, there is a need to elucidate the public health risk posed by this mosquito species in the Northeast. Accordingly, we evaluated the competence of four Ae. albopictus populations from Connecticut and New York, for two strains each of ZIKV, DENV serotype 2 (DENV-2), and CHIKV, currently circulating in the Americas, to evaluate the local transmission risk by this vector. We found that local Ae. albopictus populations are susceptible to infection by all three viruses but are most capable of transmitting CHIKV. Variation in competence was observed for ZIKV and CHIKV, driven by the virus strains and mosquito population, whereas competence was more homogeneous for the DENV-2 strains under evaluation. These results suggest that under optimal circumstances, Ae. albopictus could support localized transmission of these viruses and emphasize the importance of maintaining mosquito surveillance and control programs to suppress Ae. albopictus populations and limit further range expansion of this species.

Background: Natural infections with soil-transmitted nematodes occur in non-human primates (NHPs) and have the potential to cross primate-species boundaries and cause diseases of significant public health concern. Despite the presence of NHPs in most urban centres in Kenya, comprehensive studies on their gastrointestinal parasites are scant. Objective: Conduct a cross-sectional survey to identify zoonotic nematodes in free-ranging NHPs found within four selected urban and peri-urban centres in Kenya. Methods: A total of 86 NHPs: 41 African green monkeys [AGMs] (Chlorocebus aethiops), 30 olive baboons (Papio anubis), 5 blue monkeys (Cercopithecus mitis stuhlmanni) and 10 red-tailed monkeys (Cercopithecus ascanius) were sampled once in situ and released back to their habitat. Microscopy was used to identify nematodes egg and larvae stages in the samples. Subsequently, PCR coupled with high-resolution melting (PCR-HRM) analysis and sequencing were used to identify nodule worms. Results: NHPs inhabiting densely populated urban environs in Kenya were found infected with a rich diversity of nematodes including three potentially zoonotic nematodes including Oesophagostomum stephanostomum, Oesophagostomum bifurcum and Trichostrongylus colubriformis and co-infections were common. Conclusion: Phylogenetic analysis showed that O. stephanostomum from red-tailed and blue monkeys have a close evolutionary relatedness to human isolates suggesting the zoonotic potential of this parasite. Moreover, we also report the first natural co-infection of O. bifurcum and O. stephanostomum in free-ranging AGMs.

Aedes albopictus is a vector of arboviruses with high rates of morbidity and mortality. The northern limit of Ae. albopictus in the northeastern United States runs through New York state (NYS) and Connecticut. We present a landscape-level analysis of mosquito abundance measured by daily counts of Ae. albopictus from 338 trap sites in 12 counties during May-September 2017. During the study period, the mean number of Ae. albopictus caught per day of trapping across all sites was 3.21. We constructed four sets of negative binomial generalized linear models to evaluate how trapping methodology, land cover, as well as temperature and precipitation at multiple time intervals influenced Ae. albopictus abundance. Biogents-Sentinel (BGS) traps were 2.78 times as efficient as gravid traps and 1.49 times as efficient as CO2-baited CDC light traps. Greater proportions of low- and medium-intensity development and low proportions of deciduous cover around the trap site were positively associated with increased abundance, as were minimum winter temperature and March precipitation. The cumulative precipitation within a 28-day time window before the date of collection had a nonlinear relationship with abundance, such that greater cumulative precipitation was associated with increased abundance until approximately 70 mm, above which there was a decrease in abundance. We concluded that populations are established in Nassau, Suffolk, and New York City counties in NYS; north of these counties, the species is undergoing population invasion and establishment. We recommend that mosquito surveillance programs monitoring the northward invasion of Ae. albopictus place BGS traps at sites chosen with respect to land cover.

As new and re-emerging vector-borne diseases are occurring across the world, East Africa represents an interesting location, being the origin of several arboviruses with a history of urbanization and global spread. Rapid expansion of urban populations and alteration of natural habitats creates the opportunity for arboviruses to host-switch from wild, sylvatic hosts or vectors into urban transmission affecting human populations. Although mosquito surveillance regularly takes place in urban areas of Kenya, for example identifying vectors of dengue virus or malaria viruses, little work has been carried out to determine the distribution and abundance of sylvatic vectors. Here, we describe the mosquito vector species and diversity collected at twelve forest habitats of rural Kenya. We conducted arbovirus screening of over 14,082 mosquitoes (47 species, 11 genera) as 1520 pools, and detected seven viruses (six bunyaviruses, and one flavivirus-bunyavirus co-infection) isolated from pools of Aedes dentatus, Anopheles funestus, Culex annulioris, and Cx. vansomereni. Awareness of sylvatic vector species and their location is a critical part of understanding the ecological foci and enzootic cycling of pathogens that may be of concern to public, animal or wildlife health. As natural ecosystems come under anthropogenic pressures, such knowledge can inform us of the One Health potential for spillover or spillback leading to outbreaks, and assist in vector control strategies.

Aedes albopictus is a vector of arboviruses with high rates of morbidity and mortality. The northern limit of Ae. albopictus in the northeastern United States runs through New York state (NYS) and Connecticut. We present a landscape-level analysis of mosquito abundance measured by daily counts of Ae. albopictus from 338 trap sites in 12 counties during May-September 2017. During the study period, the mean number of Ae. albopictus caught per day of trapping across all sites was 3.21. We constructed four sets of negative binomial generalized linear models to evaluate how trapping methodology, land cover, as well as temperature and precipitation at multiple time intervals influenced Ae. albopictus abundance. Biogents-Sentinel (BGS) traps were 2.78 times as efficient as gravid traps and 1.49 times as efficient as CO2-baited CDC light traps. Greater proportions of low- and medium-intensity development and low proportions of deciduous cover around the trap site were positively associated with increased abundance, as were minimum winter temperature and March precipitation. The cumulative precipitation within a 28-day time window before the date of collection had a nonlinear relationship with abundance, such that greater cumulative precipitation was associated with increased abundance until approximately 70 mm, above which there was a decrease in abundance. We concluded that populations are established in Nassau, Suffolk, and New York City counties in NYS; north of these counties, the species is undergoing population invasion and establishment. We recommend that mosquito surveillance programs monitoring the northward invasion of Ae. albopictus place BGS traps at sites chosen with respect to land cover.

Background La Crosse virus (LACV) (genus Orthobunyavirus, family Peribunyaviridae) is a mosquito-borne virus that causes pediatric encephalitis and accounts for 50–150 human cases annually in the USA. Human cases occur primarily in the Midwest and Appalachian regions whereas documented human cases occur very rarely in the northeastern USA. Methods Following detection of a LACV isolate from a field-collected mosquito in Connecticut during 2005, we evaluated the prevalence of LACV infection in local mosquito populations and genetically characterized virus isolates to determine whether the virus is maintained focally in this region. Results During 2018, we detected LACV in multiple species of mosquitoes, including those not previously associated with the virus. We also evaluated the phylogenetic relationship of LACV strains isolated from 2005–2018 in Connecticut and found that they formed a genetically homogeneous clade that was most similar to strains from New York State. Conclusion Our analysis argues for local isolation and long-term persistence of a genetically distinct lineage of LACV within this region. We highlight the need to determine more about the phenotypic behavior of these isolates, and whether this virus lineage poses a threat to public health.