The effect of fluralaner treatment of small mammals on the endemic cycle of Borrelia burgdorferi in a natural environment.

Among approaches aimed at reducing Lyme disease risk in the environment, those targeting reservoirs of Borrelia burgdorferi Johnson are promising because they have the potential to reduce both the density of questing Ixodes scapularis Say (Acari: Ixodidea) ticks and the prevalence of B. burgdorferi in the tick population. In this 4-yr field study, we treated a population of wild small mammals with 2 densities of fluralaner baits and investigated the effect of the treatment on 3 parameters of the endemic cycle of B. burgdorferi: (i) the prevalence of infected Peromyscus mice (PIM), (ii) the density of questing nymphs (DON), and (iii) the prevalence of infected questing nymphs (NIP). We demonstrated that fluralaner baiting is effective at reducing tick infestation of Peromyscus mice, the main reservoir of B. burgdorferi in central and northeastern North America, in the laboratory and the field. Results from this study showed a significant decrease in B. burgdorferi infection in mice (odds ratio: 0.37 [CI95: 0.17 to 0.83]). A reduction in the DON between 45.4% [CI95: 22.4 to 61.6] and 62.7% [CI95: 45.9 to 74.2] occurred in treated area when compared with control areas. No significant effect was reported on the NIP. These results confirm the hypothesis that fluralaner baits have an effect on B. burgdorferi endemic cycle, with the potential to reduce the density of B. burgdorferi-infected ticks in the environment. Further studies performed in various habitats and public health intervention contexts are needed to refine and operationalize this approach for reducing Lyme disease risk in the environment.

Pharmacological studies and pharmacokinetic modelling to support the development of interventions targeting ecological reservoirs of Lyme disease.

The development of interventions targeting reservoirs of Borrelia burgdorferi sensu stricto with acaricide to reduce the density of infected ticks faces numerous challenges imposed by ecological and operational limits. In this study, the pharmacokinetics, efficacy and toxicology of fluralaner were investigated in Mus musculus and Peromyscus leucopus mice, the main reservoir of B. burgdorferi in North America. Fluralaner showed rapid distribution and elimination, leading to fast plasma concentration (Cp) depletion in the first hours after administration followed by a slow elimination rate for several weeks, resulting in a long terminal half-life. Efficacy fell below 100% while Cp (± standard deviation) decreased from 196 ± 54 to 119 ± 62 ng/mL. These experimental results were then used in simulations of fluralaner treatment for a duration equivalent to the active period of Ixodes scapularis larvae and nymphs. Simulations showed that doses as low as 10 mg/kg have the potential to protect P. leucopus against infestation for a full I. scapularis active season if administered at least once every 7 days. This study shows that investigating the pharmacology of candidate acaricides in combination with pharmacokinetic simulations can provide important information to support the development of effective interventions targeting ecological reservoirs of Lyme disease. It therefore represents a critical step that may help surpass limits inherent to the development of these interventions.

Lime for Lyme: Treatment of Leaf Litter with Dolomitic Lime Powder Impairs Activity of Immature Ixodes scapularis Ticks.

Background: Tick-borne diseases are an emerging threat to public health throughout the temperate world, leading to a growing field of research aimed at developing and testing intervention strategies for reducing human-tick encounters or prevalence of infection in ticks. Various wide-spectrum chemical acaricides have proven effective for controlling tick populations, but many of these have potential deleterious side-effects on health and the environment. In addition to chemical acaricides, certain compounds such as diatomaceous earth have been shown to have physical acaricidal properties. We hypothesized that dolomitic lime (CaMg(CO3)2, a corrosive, desiccant mineral that is already used extensively in agricultural and forestry contexts to balance the pH of soils, may affect ticks' locomotory activity, habitat position, or survival and that this should manifest as a reduction in the number of questing ticks collected by dragging. Objective: This study aimed to formally assess this hypothesis in a controlled laboratory setting. Methods: We carried out a microcosm experiment, with one control and three treated microcosm trays, each replicating the natural substrate characterizing I. scapularis habitat in northeastern North America. Each tray was infested with 200 living larvae and 50 nymphs, and then treated with 0 (control), 50, 100, or 500 g/m2 of lime powder. Ticks were collected by microdragging 24 and 72 h postliming. Results: Efficacy of liming at reducing the number of collected questing ticks ranged from 87% to 100% for larvae and 0% to 69% for nymphs 24 h postliming and from 91% to 93% for larvae and -47% to 65% for nymphs 72 postliming. Conclusion: This study provides the first experimental evidence of the potential efficacy of liming for impairing activity of questing immature ticks. Given that lime is a low-cost material, that methods for widespread application in deciduous woodlands already exist, and that it has been documented as having a limited negative impact on the environment, further assessment of lime application as a public health risk reduction intervention for tick-borne diseases is warranted.

The added value of One Health surveillance: data from questing ticks can provide an early signal for anaplasmosis outbreaks in animals and humans.

OBJECTIVE: In 2021, a first outbreak of anaplasmosis occurred in animals and humans in southern Québec, with 64% of confirmed human cases located in Bromont municipality. Ixodes scapularis ticks and Peromyscus mouse ear biopsies collected in Bromont from 2019 to 2021 were analyzed for Anaplasma phagocytophilum (Ap) with the objective of determining whether an early environmental signal could have been detected before the outbreak. METHODS: Samples were collected for a concurrent study aiming to reduce Lyme disease risk. Between 2019 and 2021, up to 14 experimental sites were sampled for ticks and capture of small mammals took place on three sites in 2021. Samples were screened for Ap using multiplex real-time PCR, and genetic strains were identified using a single-nucleotide polymorphism assay. RESULTS: Analyses showed an increase of 5.7% in Ap prevalence in ticks (CI95: 1.5-9.9) between 2019 and 2020, i.e., one year before the outbreak. A majority of Ap-positive ticks were infected with the zoonotic strain (68.8%; CI95: 50.0-83.9) during the study period. In 2021, 2 of 59 captured Peromycus mice were positive for Ap, for a prevalence of 3.4% (CI95: 0.4-11.7). CONCLUSION: We conclude that data collected in Bromont could have provided an early signal for an anaplasmosis risk increasing in the targeted region. This is a reminder that integrated surveillance of tick-borne diseases through structured One Health programs, i.e. systematically integrating data from humans, animals and the environment, can provide useful and timely information for better preparedness and response in public health.

RéSUMé: OBJECTIF: En 2021, suivant une éclosion d'anaplasmoses chez les animaux et les humains dans le sud du Québec, des tiques de l'espèce Ixodes scapularis et des biopsies de souris Peromyscus spp. échantillonées à Bromont, la municipalité où 64 % des cas humains confirmés était localisé, ont été testées pour Anaplasma phagocytophilum (Ap) avec pour objectif de déterminer si un signal environnemental précoce d'augmentation du risque aurait pu être détecté avant l'éclosion. MéTHODE: L'échantillonnage a été réalisé dans le cadre d'une étude visant à réduire le risque de maladie de Lyme. De 2019 à 2021, 14 sites expérimentaux ont été échantillonnés pour les tiques. En 2021, trois sites ont été sélectionnés pour la capture des micromammifères. Les échantillons ont été testés pour la présence d'Ap à l'aide d'un PCR multiplex en temps réelle et les lignées génétiques ont été identifiées grâce à un test de polymorphisme mononucléotidique. RéSULTATS: Les analyses ont montré une augmentation de 5,7 % (IC95% : 1,5­9,9) de la prévalence de Ap entre 2019 et 2020, c'est-à-dire un an avant l'éclosion. Cette augmentation est associée à la présence d'une majorité d'Ap de la lignée zoonotique (68,8 %; IC95% : 50,0­83,9) sur l'ensemble de la période étudiée. En 2021, deux Peromycus spp. capturées sur 59 étaient positives pour Ap pour une prévalence de 3,4 % (IC95% : 0,4­11,7). CONCLUSION: Les données environnementales échantillonnées à Bromont auraient pu fournir un signal précoce de l'augmentation du risque d'anaplasmose dans la région. C'est un rappel que la surveillance intégrée des maladies transmises par les tiques inspirée de l'approche Une seule santé, intégrant systématiquement des données humaines, animales et environnementales, peut fournir des informations utiles et opportunes aux autorités de santé publique.

Fluralaner Baits Reduce the Infestation of Peromyscus spp. Mice (Rodentia: Cricetidae) by Ixodes scapularis (Acari: Ixodidae) Larvae and Nymphs in a Natural Environment.

The development of interventions that reduce Lyme disease incidence remains a challenge. Reservoir-targeted approaches aiming to reduce tick densities or tick infection prevalence with Borrelia burgdorferi have emerged as promising ways to reduce the density of infected ticks. Acaricides of the isoxazoline family offer high potential for reducing infestation of ticks on small mammals as they have high efficacy at killing feeding ticks for a long period. Fluralaner baits were recently demonstrated as effective, in the laboratory, at killing Ixodes scapularis larvae infesting Peromyscus mice, the main reservoir for B. burgdorferi in northeastern North America. Here, effectiveness of this approach for reducing the infestation of small mammals by immature stages of I. scapularis was tested in a natural environment. Two densities of fluralaner baits (2.1 baits/1,000 m2 and 4.4 baits/1,000 m2) were used during three years in forest plots. The number of I. scapularis larvae and nymphs per mouse from treated and control plots were compared. Fluralaner baiting reduced the number of larvae per mouse by 68% (CI95: 51-79%) at 2.1 baits/1,000 m2 and by 86% (CI95: 77-92%) at 4.4 baits/1,000 m2. The number of nymphs per mouse was reduced by 72% (CI95: 22-90%) at 4.4 baits/1,000 m2 but was not significantly reduced at 2.1 baits/1,000 m2. Reduction of Peromyscus mouse infestation by immature stages of I. scapularis supports the hypothesis that an approach targeting reservoirs of B. burgdorferi with isoxazolines has the potential to reduce tick-borne disease risk by decreasing the density of infected ticks in the environment.

Can local risk of West Nile virus infection be predicted from previous cases? A descriptive study in Quebec, 2011-2016.

OBJECTIVES: This study aimed at (1) describing the local risk of West Nile virus (WNV) infection in humans based on previous case reports and (2) investigating the spatial clustering of cases in the five most affected administrative regions of Quebec, Canada, for the 2011-2016 period. METHODS: Human WNV cases declared to the Ministry of Health and Social Services of Quebec (Ministère de la santé et des services sociaux, MSSS) were retrieved. Incidence risk by age and sex was calculated for the study period. The yearly and monthly occurrence of cases in geographical units (GUs) was described and the probability of observing cases in a GU with cases reported in the previous year or month was assessed. Moran's I was used to assess global clustering across the study area. Spatial clusters were identified by the Kulldorff scan statistic. RESULTS: A total of 261 WNV cases were declared to the MSSS between 2011 and 2016 in the study area. Overall, a low percentage of GU with cases reported had additional cases reported over the next month or year. Global spatial clustering was weak but statistically significant (p < 0.05) for 2012 and 2015. For these two years, spatial clusters of high-risk GUs were identified. CONCLUSION: Results underline the challenge of predicting the distribution of WNV incidence risk in Quebec based on previous occurrence of human cases. Ongoing research with high spatial resolution entomological data is still necessary to understand the spatial distribution of risk at a local scale.

Evaluation of fluralaner as an oral acaricide to reduce tick infestation in a wild rodent reservoir of Lyme disease.

BACKGROUND: Lyme disease (LD) is an increasing public health threat in temperate zones of the northern hemisphere, yet relatively few methods exist for reducing LD risk in endemic areas. Disrupting the LD transmission cycle in nature is a promising avenue for risk reduction. This experimental study evaluated the efficacy of fluralaner, a recent oral acaricide with a long duration of effect in dogs, for killing Ixodes scapularis ticks in Peromyscus maniculatus mice, a known wildlife reservoir for Borrelia burgdorferi in nature. METHODS: We assigned 87 mice to 3 fluralaner treatment groups (50 mg/kg, 12.5 mg/kg and untreated control) administered as a single oral treatment. Mice were then infested with 20 Ixodes scapularis larvae at 2, 28 and 45 days post-treatment and we measured efficacy as the proportion of infesting larvae that died within 48 h. At each infestation, blood from 3 mice in each treatment group was tested to obtain fluralaner plasma concentrations (Cp). RESULTS: Treatment with 50 mg/kg and 12.5 mg/kg fluralaner killed 97% and 94% of infesting larvae 2 days post-treatment, but no significant effect of treatment on feeding larvae was observed 28 and 45 days post-treatment. Mouse Cp did not differ significantly between the two tested doses. Mean Cp decreased from 13,000 ng/ml in the 50 mg/kg group and 4000 ng/ml in the 12.5 mg/kg group at Day 2 to < 100 ng/ml in both groups at Day 45. CONCLUSIONS: We provide the first evidence that fluralaner is effective for killing immature ticks in Peromyscus mice, a first step in evaluating its potential for treating wild rodents as a public health intervention to reduce LD risk in endemic areas.

Eastern equine encephalitis virus: high seroprevalence in horses from Southern Quebec, Canada, 2012.

Eastern equine encephalitis virus (EEEV) is a highly pathogenic arbovirus that infects humans, horses, and other animals. There has been a significant increase in EEEV activity in southeastern Canada since 2008. Few data are available regarding nonlethal EEEV infections in mammals, and consequently the distribution and pathogenicity spectrum of EEEV infections in these hosts is poorly understood. This cross-sectional study focuses on the evaluation of viral activity in southern Quebec's horses by seroprevalence estimation. A total of 196 horses, 18 months and older, which had never been vaccinated against EEEV and have never traveled outside Canada, were sampled from 92 barns distributed throughout three administrative regions of southern Quebec. Blood samples were taken from each horse and titrated for EEEV antibodies by plaque reduction neutralization test (PRNT). Equine population vaccination coverage was estimated by surveying horse owners and equine practitioners. PRNT results revealed an EEEV seroprevalence up to 8.7%, with 95% confidence limits ranging from 4.4% to 13.0%. Vaccination coverage was estimated to be at least 79%. Our study reveals for the first time in Canada a measure of EEEV seroprevalence in horses. High seroprevalence in unvaccinated animals challenges the perception that EEEV is a highly lethal pathogen in horses. Monitoring high-risk vector-borne infections such as EEEV in animal populations can be an important element of a public health surveillance strategy, population risk assessment and early detection of epidemics.