Male C57BL/6J mice have higher presence and abundance of Borrelia burgdorferi in their ventral skin compared to female mice.

Borrelia burgdorferi is a tick-borne spirochete that causes Lyme disease in humans. The host immune system controls the abundance of the spirochete in the host tissues. Recent work with immunocompetent Mus musculus mice strain C3H/HeJ found that males had a higher tissue infection prevalence and spirochete load compared to females. The purpose of this study was to determine whether host sex and acquired immunity interact to influence the prevalence and abundance of spirochetes in the tissues of the commonly used mouse strain C57BL/6. Wildtype (WT) mice and their SCID counterparts (C57BL/6) were experimentally infected with B. burgdorferi via tick bite. Ear biopsies were sampled at weeks 4, 8, and 12 post-infection (PI) and five tissues (left ear, ventral skin, heart, tibiotarsal joint of left hind leg, and liver) were collected at necropsy (16 weeks PI). The mean spirochete load in the tissues of the SCID mice was 260.4x higher compared to the WT mice. In WT mice, the infection prevalence in the ventral skin was significantly higher in males (40.0 %) compared to females (0.0 %), and the spirochete load in the rear tibiotarsal joint was significantly higher (4.3x) in males compared to females. In SCID mice, the spirochete load in the ventral skin was 200.0x higher in males compared to females, but there were no significant sex-specific difference in spirochete load in the other tissues (left ear, heart, tibiotarsal joint, or liver). Thus, the absence of acquired immunity greatly amplified the spirochete load in the ventral skin of male mice. It is important to note that the observed sex-specific differences in laboratory mice cannot be extrapolated to humans. Future studies should investigate the mechanisms underlying the male bias in the abundance of B. burgdorferi in the mouse skin.

The influence of weather on the population dynamics of common mosquito vector species in the Canadian Prairies.

BACKGROUND: Mosquito seasonal activity is largely driven by weather conditions, most notably temperature, precipitation, and relative humidity. The extent by which these weather variables influence activity is intertwined with the animal's biology and may differ by species. For mosquito vectors, changes in weather can also alter host-pathogen interactions thereby increasing or decreasing the burden of disease. METHODS: In this study, we performed weekly mosquito surveillance throughout the active season over a 2-year period in Manitoba, Canada. We then used Generalized Linear Mixed Models (GLMMs) to explore the relationships between weather variables over the preceding 2 weeks and mosquito trap counts for four of the most prevalent vector species in this region: Oc. dorsalis, Ae. vexans, Cx. tarsalis, and Cq. perturbans. RESULTS: More than 265,000 mosquitoes were collected from 17 sampling sites throughout Manitoba in 2020 and 2021, with Ae. vexans the most commonly collected species followed by Cx. tarsalis. Aedes vexans favored high humidity, intermediate degree days, and low precipitation. Coquillettidia perturbans and Oc. dorsalis activity increased with high humidity and high rainfall, respectively. Culex tarsalis favored high degree days, with the relationship between number of mosquitoes captured and precipitation showing contrasting patterns between years. Minimum trapping temperature only impacted Ae. vexans and Cq. perturbans trap counts. CONCLUSIONS: The activity of all four mosquito vectors was affected by weather conditions recorded in the 2 weeks prior to trapping, with each species favoring different conditions. Although some research has been done to explore the relationships between temperature/precipitation and Cx. tarsalis in the Canadian Prairies, to our knowledge this is the first study to investigate other commonly found vector species in this region. Overall, this study highlights how varying weather conditions can impact mosquito activity and in turn species-specific vector potential.

Clobetasol increases the abundance of Borrelia burgdorferi in the skin 70 times more in male mice compared to female mice.

Lyme borreliosis is caused by the spirochete Borrelia burgdorferi and is transmitted among vertebrate hosts by Ixodes scapularis ticks in eastern North America. Treatment with topical corticosteroids increases the abundance of B. burgdorferi in the skin of lab mice that have been experimentally infected via needle inoculation. In the present study, female and male C3H/HeJ mice were infected with B. burgdorferi via nymphal tick bite. Infected mice were treated with clobetasol on the skin of the right hindleg on days 35 and 36 post-infection and euthanized at days -2, 1, 3, 5, and 7 post-treatment; a group of control mice was infected but not treated with clobetasol. The spirochete abundance was quantified in 8 mouse tissues including bladder, heart, left hindleg skin, right hindleg skin, dorsal skin, ventral skin, left ear and right ear. Averaged across the 8 mouse tissues, the abundance of B. burgdorferi on days 3 and 5 were 21.4x and 14.4x higher in mice treated with clobetasol compared to the untreated control mice, but there were large differences among tissues. There was a dramatic sex-specific effect of the clobetasol treatment; the peak abundance of B. burgdorferi in the skin (left hindleg, right hindleg, dorsal, ventral) was 72.6x higher in male mice compared to female mice. In contrast, there was little difference between the sexes in the tissue spirochete load in the ears, bladder, and heart. Topical application of clobetasol could increase the sensitivity of direct diagnostic methods (e.g., culture, PCR) to detect B. burgdorferi in host skin biopsies.

Transcriptional profiling of Dermacentor variabilis (Acari: Ixodidae) provides insights into the role of the Haller's organ in spatial DEET recognition.

DEET is the most common active ingredient in commercial repellents, providing effective protection against blood-sucking insects and ticks. However, its mode of action is not fully understood, with several theories put forward to explain its repellency effect. Unique to ticks, the Haller's organ recognizes a variety of external stimuli through non-contact mechanisms, yet the extent to which the organ plays a role in tick chemoreception is not fully known. We previously found that DEET inhibited the expression/activity of cytochrome P450s and cholinesterases in Dermacentor variabilis, however, our experimental design could not distinguish between sexes (males/females), method of exposure (volatile/tactile) or the roles of chemosensory tissues (Haller's organ). In this study, we used RNA sequencing to assess changes in transcript expression induced by volatile DEET in D. variabilis males/females with/without intact Haller's organs. Male ticks showed much greater transcriptional responses to DEET than females, which may be at least partially attributed to the sexual dimorphism of the Haller's organ. Female transcript expression profiles were most influenced by condition (i.e., intact/excised Haller's organs) with minimal changes due to repellent exposure. On the other hand, removal of the Haller's organs caused DEET treated male ticks to exhibit similar expression profiles as control (ethanol) ticks with intact Haller's organs. Consequently, the transcript-level responses to spatial DEET exposure appears largely based on males possessing their Haller's organs. The molecular signature of this response included the suppression of a large number of transcripts involved in detoxification, lipid metabolism and immunity. Taken collectively, this study furthers our understanding of the Haller's organ role in volatile DEET recognition.

Three years of surveillance associates agro-environmental factors with wireworm infestations in Manitoba, Canada.

BACKGROUND: Wireworms, the soil-dwelling larvae of click beetles, are a major threat to global agricultural production. This is largely due to their generalist polyphagous feeding capabilities, extended and cryptic life cycles, and limited management options available. Although wireworms are well-documented as economically important pests in the Canadian Prairies, including Manitoba, there are gaps in knowledge on species distributions, subterranean behaviour and life cycles, feeding ecology and damage capacity, and economic thresholds for crop yield loss. RESULTS: We carried out 3 years (2018-2020) of intensive surveillance of larval populations across Manitoba. A total of 31 fields (24 in ≥ 2 consecutive years) were surveyed in early spring using standardized bait trapping approaches. Wireworms were present in 94% of surveyed sites, but the catch within fields varied year to year. While Hypnoidus bicolor predominated (94% of larvae), several other pest species were identified. We then explored the relationships between wireworm trap numbers and agro-environmental factors. The larval catch tended to decrease under conditions of low soil temperatures and increased clay content, coupled with high soil moisture and precipitation during the trapping period. Treatment and cultural methods appeared less influential; however, wheat production in either of the previous two growing seasons was associated with increased wireworm catch. Our models failed to predict a relationship between wireworm catch and crop yields, although infestations were rare in our region. CONCLUSION: Our findings better infer the risks posed by wireworms to crop production in the Canadian Prairies, and the agro-environmental factors that represent the greatest contributors to these risks. This information should be incorporated into future integrated pest management (IPM) strategies for wireworms. © 2021 Her Majesty the Queen in Right of Canada Pest Management Science © 2021 Society of Chemical Industry Reproduced with the permission of the Minister of Agriculture and Agri-Food Canada.

Occurrence of the Mosquito Aedes triseriatus (Diptera: Culicidae) Beyond Its Most Northwestern Range Limits in Manitoba, Canada.

Native to the Eastern United States and Eastern Canada, Aedes triseriatus (eastern tree hole mosquito) is an important vector of La Crosse virus and dog heartworm. Although its range has been well characterized in the United States, few studies have surveyed its distribution within Canada. In this study, mosquitoes were collected from a variety of urban and rural communities throughout Manitoba, Canada between the years of 2018 and 2020. Aedes triseriatus was identified and confirmed molecularly to be present in 13 communities. This includes localities that expand the species known distribution to new northern and western areas, and suggests that past surveillance efforts have not been comprehensive or environmental factors have caused this mosquito species to be present in areas in which it was not found previously. As Canada is showing signs of a changing climate, this may be driving the broader occurrence of Ae. triseriatus.

Genetic structure and population demographics of Hypnoidus bicolor (Coleoptera: Elateridae) in the Canadian Prairies.

BACKGROUND: Following banning of the pesticide lindane in most counties, wireworms (i.e., the soil-living larval stages of click beetles) have become major pests of a variety of economically important field crops. Hypnoidus bicolor is a common pest species in the Canadian Prairies. However, little is known about its life history, which impedes the development of effective integrated pest management (IPM) strategies. Population genetic approaches have the potential to assist in the development of IPM. RESULTS: We sequenced a 622-bp fragment of the COX1 gene from 326 H. bicolor wireworm and click beetles collected from 13 localities on the Canadian Prairies. Two genetically distinct (>4.66% sequence divergence) clades were identified, suggesting that they may be part of a species complex. Clade A predominated and increased in prevalence the further east samples were collected, whereas the opposite was true for clade B. Clade B appears to be comprised of two mitochondrial DNA groups, however, one group was represented by only one haplotype. Both clades were characterized by uneven gene flow among populations with low levels of regional genetic structuring. Clade A appeared to have undergone population and range expansions, which may coincide with the advent of intensive agriculture practices in the prairies. CONCLUSION: Knowledge of species composition and population structure is important for the development of effective IPM strategies but is often lacking for wireworms. Our study fills these knowledge gaps for a predominant pest species in the prairies, H. bicolor, by providing robust evidence for cryptic forms and characterizing its dispersal patterns and population dynamics. © 2021 Society of Chemical Industry.

Natural insensitivity and the effects of concentration on the repellency and survival of American dog ticks (Dermacentor variabilis) by DEET.

N,N-Diethyl-m-toluamide (DEET) is by far the most used repellent worldwide. When applied topically to the skin, the active ingredient has been shown to provide protection from a variety of hematophagous insects, including mosquitoes and flies. DEET's effectiveness against ticks is influenced by a variety of factors (e.g., duration and concentration of application, drying time, route of exposure, tick species and developmental stage), and may differ from insects due to their unique chemosensory system that primarily involves the Haller's organ. We therefore used several approaches to investigate DEET's efficacy to repel Dermacentor variabilis at different concentrations (5, 30 or 75%), as well as explore its toxicological properties and natural variability in DEET insensitivity across populations from Manitoba, Canada. Climbing bioassays indicated that higher concentrations of DEET were more effective at repelling D. variabilis, and that ticks from some sampling localities were more sensitive to lower concentrations than others. Petri dish arena assays revealed ticks exposed to high concentrations of the repellent lose their ability to discriminate lower concentrations, perhaps due to overstimulation or habituation. Finally, our tactile assays demonstrated reduced tick survival after contact with high DEET concentrations, with mortality occurring more rapidly with increased concentration. Dermacentor variabilis from these tactile assays displayed a multitude of physiological and neurological symptoms, such as 'hot foot' and various bodily secretions. Overall, our study shows a strong association between repellency, concentration and the acaricidal effects of DEET on D. variabilis.

Molecular evidence for the inhibition of cytochrome p450s and cholinesterases in ticks by the repellent DEET.

For more than 50 years DEET (N,N-Diethyl-m-toluamide) has been considered the gold standard of repellents. It is applied to the skin or clothing to deter mosquitoes and other blood-sucking invertebrate pests from approaching and/or settling, and ultimately it provides temporary protection from bites. Despite rampant global use, surprisingly little is understood about DEET's mode of action and the molecular targets of the active ingredient. Furthermore, the theories into its mechanism for repellency are largely based off fruit fly and mosquito research. Since ticks possess a unique sensory structure, the Haller's organ, the specific genes and pathways associated with DEET avoidance may differ from insects. In these studies, we collected American dog ticks (Dermacentor variabilis) from four natural populations within Manitoba, Canada. We first carried out behavior assays, which showed DEET effectively repelled the ticks. RNA sequencing revealed that DEET caused a rapid and substantial reduction in the abundance of transcripts encoding cytochrome P450 and acetylcholinesterase genes, which gradually recovered over the 24 h time course. Finally, enzymatic kinetics provided functional support for DEET's role as an effective inhibitor of P450 s. While many facets of its mode of action remain to be worked out, our study provides valuable insights into the molecular underpinnings of DEET's repellence in ticks.