Meteorological and climatic variables predict the phenology of Ixodes ricinus nymph activity in France, accounting for habitat heterogeneity.

Ixodes ricinus ticks (Acari: Ixodidae) are the most important vector for Lyme borreliosis in Europe. As climate change might affect their distributions and activities, this study aimed to determine the effects of environmental factors, i.e., meteorological, bioclimatic, and habitat characteristics on host-seeking (questing) activity of I. ricinus nymphs, an important stage in disease transmissions, across diverse climatic types in France over 8 years. Questing activity was observed using a repeated removal sampling with a cloth-dragging technique in 11 sampling sites from 7 tick observatories from 2014 to 2021 at approximately 1-month intervals, involving 631 sampling campaigns. Three phenological patterns were observed, potentially following a climatic gradient. The mixed-effects negative binomial regression revealed that observed nymph counts were driven by different interval-average meteorological variables, including 1-month moving average temperature, previous 3-to-6-month moving average temperature, and 6-month moving average minimum relative humidity. The interaction effects indicated that the phenology in colder climates peaked differently from that of warmer climates. Also, land cover characteristics that support the highest baseline abundance were moderate forest fragmentation with transition borders with agricultural areas. Finally, our model could potentially be used to predict seasonal human-tick exposure risks in France that could contribute to mitigating Lyme borreliosis risk.

Habitat suitability map of Ixodes ricinus tick in France using multi-criteria analysis.

The tick Ixodes ricinus is widely distributed across Europe and is responsible for the transmission of several pathogens to humans and animals. In this study, we used a knowledge-based method to map variations in habitat suitability for I. ricinus ticks throughout continental France and Corsica. The multi-criteria decision analysis (MCDA) integrated four major biotic and abiotic factors known to influence tick populations: climate, land cover, altitude and the density of wild ungulates. For each factor, habitat suitability index (HSI) values were attributed to different locations based on knowledge regarding its impact on tick populations. For the MCDA, two methods of factor combination were tested, additive and multiplicative, both which were evaluated at the spatial scales of departments and local municipalities. The resulting habitat suitability maps (resolution=100x100 m) revealed that conditions are suitable for I. ricinus over most of France and Corsica. Particularly suitable habitats were located in central, north-eastern and south-western France, while less-suitable habitats were found in the Mediterranean and mountainous regions. To validate the approach, the HSI scores were compared to field data of I. ricinus nymph abundance. Regardless of scale, the correlation between abundance indicator and HSI score was stronger for the additive than for the multiplicative approach. Overall, this study demonstrates the value of MCDA for estimating habitat suitability maps for I. ricinus abundance, which could be especially useful in highlighting areas of the tick's distribution where preventive measures should be prioritised.

Tick ecology and Lyme borreliosis prevention: a regional survey of pharmacists' knowledge in Auvergne-Rhône-Alpes, France.

The most prevalent vector-borne diseases in Europe are caused by tick-borne pathogens, such as bacteria of the genus Borrelia that cause Lyme borreliosis. In this context, retail pharmacists are frequently the first medical source of information in the event of a tick bite. The objective of this study was to assess pharmacy professionals' knowledge about both tick ecology and the appropriate measures for tick bites and Lyme borreliosis prevention. It was based on an online survey of 364 pharmacists and pharmacy assistants located in the Auvergne-Rhône-Alpes region of France. The results showed solid knowledge about preventive measures for tick bite and Lyme borreliosis, but weaker knowledge about tick biology (hosts, suitable habitats, favorable conditions for tick activity, etc.). In particular, several stereotypes were observed in the responses of the pharmacy professionals. These appear to result from a social construction of the knowledge on ticks and tick-borne diseases previously shown to the general population in the region. The results highlight the need for continuous training about ticks and tick-borne diseases for healthcare professionals serving local populations that live in endemic areas.

Questing tick abundance in urban and peri-urban parks in the French city of Lyon.

BACKGROUND: In Europe, ticks are responsible for the transmission of several pathogens of medical importance, including bacteria of the Borrelia burgdorferi (s.l.) complex, the agents of Lyme borreliosis. In France, the Auvergne Rhône-Alpes region is considered a hot spot for human tick-borne pathogen infections, with an estimated annual rate of 156 cases of Lyme borreliosis per 100,000 inhabitants. Although several studies have assessed the abundance of ticks in rural areas, little consideration has been given thus far to urban green spaces in France. METHODS: This study aimed to estimate tick abundance in three parks, two urban (U1, U2) and one peri-urban (PU), in and around the city of Lyon (France). A forest in a rural area was used as a control (C). Tick sampling campaigns were performed in each site in April, May, June, July, and October 2019 using the dragging method. One hundred transects of 10 m2 each were randomly chosen in each park in places frequented by humans. The sampling sessions were carried out under semi-controlled abiotic conditions. Ticks were stored in 70% ethanol and identified to species and developmental stage under a light microscope using morphological keys. RESULTS: A total of seven ticks (nymphs and adults) were collected in the two urban parks (six in U1 and one in U2), while 499 ticks were sampled in the peri-urban park. Of the 506 ticks collected, 504 were identified as Ixodes ricinus, one as Dermacentor marginatus, and one as Ixodes frontalis. In the peri-urban park, ticks were mainly collected under the forest cover and at forest edges. Tick density under forest cover was 7.1 times higher in the control site than in the peri-urban park throughout the survey period. CONCLUSIONS: This study confirmed the presence of ticks in all of the parks surveyed, although their occurrence in the urban parks was very rare compared to the peri-urban park and the control site. These results should serve as a basis for the implementation of preventive measures.

A cautionary note on Bayesian estimation of population size by removal sampling with diffuse priors.

We consider the problem of estimating a population size by removal sampling when the sampling rate is unknown. Bayesian methods are now widespread and allow to include prior knowledge in the analysis. However, we show that Bayes estimates based on default improper priors lead to improper posteriors or infinite estimates. Similarly, weakly informative priors give unstable estimators that are sensitive to the choice of hyperparameters. By examining the likelihood, we show that population size estimates can be stabilized by penalizing small values of the sampling rate or large value of the population size. Based on theoretical results and simulation studies, we propose some recommendations on the choice of the prior. Then, we applied our results to real datasets.

Multiple independent transmission cycles of a tick-borne pathogen within a local host community.

Many pathogens are maintained by multiple host species and involve multiple strains with potentially different phenotypic characteristics. Disentangling transmission patterns in such systems is often challenging, yet investigating how different host species contribute to transmission is crucial to properly assess and manage disease risk. We aim to reveal transmission cycles of bacteria within the Borrelia burgdorferi species complex, which include Lyme disease agents. We characterized Borrelia genotypes found in 488 infected Ixodes ricinus nymphs collected in the Sénart Forest located near Paris (France). These genotypes were compared to those observed in three sympatric species of small mammals and network analyses reveal four independent transmission cycles. Statistical modelling shows that two cycles involving chipmunks, an introduced species, and non-sampled host species such as birds, are responsible for the majority of tick infections. In contrast, the cycle involving native bank voles only accounts for a small proportion of infected ticks. Genotypes associated with the two primary transmission cycles were isolated from Lyme disease patients, confirming the epidemiological threat posed by these strains. Our work demonstrates that combining high-throughput sequence typing with networks tools and statistical modeling is a promising approach for characterizing transmission cycles of multi-host pathogens in complex ecological settings.

Reproducibility of local environmental factors for the abundance of questing Ixodes ricinus nymphs on pastures.

In ecology and epidemiology, exploratory field studies based on multivariate statistical models commonly are used to identify factors that are associated with a phenomenon. The challenge is to evaluate whether these factors are indeed correlated to the phenomenon or if the statistical significance results from fortuitous association or type 1 statistical error (probability of rejecting a null hypothesis when it is true). This is particularly the case when the phenomenon is linked to environmental factors that usually are more or less correlated to each other and when the phenomenon is itself highly variable. The abundance of Ixodes ricinus ticks, a major disease vector in Europe, is highly variable and depends on environmental conditions that define suitable habitat and host presence. Our objective was to identify reproducible factors in space and time that influenced the abundance of questing I. ricinus nymphs. We sampled questing nymphs in pastures in 4 settings, i.e. during 3 periods (2003, 2004, and 2006) in one region, and during one period (2006) in another region, both regions located in Central France. The same data collection, data selection, and model analysis using negative binomial distribution were applied independently in the 4 data sets to identify 'reproducible' factors, i.e. explanatory factors that were significant in different time periods and spaces. The 3 most reproducible factors suggested that woodland type vegetation and woodland vicinity constantly favoured nymph abundance on pastures. In addition, the presence of fruit trees was significantly associated with nymph abundance in one region. The other factors were not reproducible. The study confirmed the status of key factors for nymph abundance while avoiding having to redefine the statistical model to model the different sampling conditions. It also shows the difficulty to identify factors with general significance acting on a very variable phenomenon, based on a study made one year in one region. Relevant study design should consider appropriate repetitions based on the range of values of the studied factors. Furthermore, discussion of the results is essential to identify factors that have a general significance.

Ecological factors characterizing the prevalence of bacterial tick-borne pathogens in Ixodes ricinus ticks in pastures and woodlands.

Ecological changes are recognized as an important driver behind the emergence of infectious diseases. The prevalence of infection in ticks depends upon ecological factors that are rarely taken into account simultaneously. Our objective was to investigate the influences of forest fragmentation, vegetation, adult tick hosts, and habitat on the infection prevalence of three tick-borne bacteria, Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, and Rickettsia sp. of the spotted fever group, in questing Ixodes ricinus ticks, taking into account tick characteristics. Samples of questing nymphs and adults were taken from 61 pastures and neighboring woodlands in central France. The ticks were tested by PCR of pools of nymphs and individual adults. The individual infection prevalence was modeled using multivariate regression. The highest infection prevalences were found in adult females collected in woodland sites for B. burgdorferi sensu lato and A. phagocytophilum (16.1% and 10.7%, respectively) and in pasture sites for Rickettsia sp. (8.7%). The infection prevalence in nymphs was lower than 6%. B. burgdorferi sensu lato was more prevalent in woodlands than in pastures. Forest fragmentation favored B. burgdorferi sensu lato and A. phagocytophilum prevalence in woodlands, and in pastures, the B. burgdorferi sensu lato prevalence was favored by shrubby vegetation. Both results are probably because large amounts of edges or shrubs increase the abundance of small vertebrates as reservoir hosts. The Rickettsia sp. prevalence was maximal on pasture with medium forest fragmentation. Female ticks were more infected by B. burgdorferi sensu lato than males and nymphs in woodland sites, which suggests an interaction between the ticks and the bacteria. This study confirms the complexity of the tick-borne pathogen ecology. The findings support the importance of small vertebrates as reservoir hosts and make a case for further studies in Europe on the link between the composition of the reservoir host community and the infection prevalence in ticks.

Blood-feeding behavior of Aedes albopictus, a vector of Chikungunya on La Réunion.

Chikungunya virus (CHIKV) has long been considered to be transmitted to humans by the human-biting mosquito Aedes aegypti, especially in Africa. However, the recent outbreak of CHIKV involved another vector, Aedes albopictus, and serological data in the literature suggest that several species of domestic or human-related vertebrates can be contaminated by this virus. However, the role of Ae. albopictus mosquitoes as potential enzootic vectors for CHIKV has not yet been evaluated. Here we investigate Ae. albopictus feeding and resting behaviors in an area where a CHIKV epidemic recently occurred, which means deciphering host-seeking and feeding behaviors on several vertebrate species, measuring endophagous/exophagous (activity), endophilic/exophilic (resting) behaviors and its diel (24 h, day/night) biting activity. Ae. albopictus was found to have bimodal daily feeding activities and was found to have exophagic (89%) and exophilic (87%) behaviors. Ae. albopictus showed an opportunistic feeding behavior on a wide range of hosts (from cold-blooded to warm-blooded animals), supporting that it can be implicated in various vertebrate-virus pathosystems. However, with equal availability of one of the four vertebrate hosts (calf, chicken, dog, and goat) proposed against human, Ae. albopictus significantly preferred human, supporting earlier data about its high degree of anthropophily. Multiple blood feeding was also reported in every combination (animal/human) offered to Ae. albopictus, enlightening the higher risks to spread an arbovirus to human population because of interrupted feeding. Such catholic behavior suggests that Ae. albopictus may act as a bridge vector for zoonotic viruses. Further epidemiological implications of this issue are discussed.

No antioxidant beneficial effect of zinc supplementation on oxidative stress markers and antioxidant defenses in middle-aged and elderly subjects: the Zenith study.

OBJECTIVE: The aim of the study was to investigate whether zinc supplementation affects antioxidant status in European middle-aged and elderly people. DESIGN: Multicentre prospective intervention study, randomized, double-blind, placebo-control. SETTING: France (Clermont-Ferrand/Theix, and Grenoble), Italy (Rome), Northern Ireland (Coleraine). SUBJECTS: A total of 387 healthy middle-aged (55-70 yrs) and free-living older aged (70-85 yrs) subjects were randomly allocated to three groups: 0, 15 or 30 mg zinc gluconate/d in addition to usual dietary intake during 6 months. METHODS: Oxidative stress status was evaluated by measurement of protein oxidation (plasma thiol groups), lipid peroxidation (plasma thio-barbituric acid reactants, TBARS), whole blood glutathione levels, erythrocyte copper/zinc superoxide dismutase activity and plasma antioxidant status (ferric reducing antioxidant power assay), at baseline and after 3 and 6 months. RESULTS: Zinc supplementation did not alter oxidative stress markers and antioxidant defenses in elderly, after 3 or 6 months, except an increase in Cu/Zn superoxide dismutase activity. CONCLUSIONS: In apparently healthy free living elderly people, a single zinc supplementation had no effects on oxidative stress status.

Borrelia burgdorferi Sensu Lato in Siberian chipmunks (Tamias sibiricus) introduced in suburban forests in France.

Numerous vertebrate reservoirs have been described for Borrelia burgdorferi sensu lato (sl), which includes the etiological agents of Lyme Borreliosis (LB). The Siberian chipmunk (Tamias sibiricus) is a rodent originating from Asia, where it is suspected to be a B. burgdorferi reservoir. It has been intentionally released into the wild in Europe since the 1970s, but has not yet been subject to any study regarding its association with the LB agent. In this paper we studied Siberian chipmunk infestation with the LB vector (Ixodes ricinus) and infection prevalence by LB spirochetes in a suburban introduced population. We compared these findings with known competent reservoir hosts, the bank vole (Myodes [clethrionomys] glareolus) and wood mouse (Apodemus sylvaticus). All Siberian chipmunks were infested with larvae and larval abundance was higher in this species (mean number of larvae [95% Confidence Interval]: 73.5 [46.0, 117.2]) than in the two other rodent species (bank voles: 4.4 [3.0, 6.3] and wood mice: 10.2 [4.9, 21.2]). Significant factors affecting abundance of larvae were host species and sampling season. Nymphs were most prevalent on chipmunks (86.2%, mean: 5.1 [3.3, 8.0]), one vole carried only two nymphs, and none of the mice had any nymphs. Nymph abundance in chipmunks was affected by sampling season and sex. Furthermore, the infection prevalence of B. burgdorferi sl in the Siberian chipmunk was the highest (33.3%) and predominantly of B. afzelii. The infection prevalence was 14.1% in bank voles, but no wood mouse was found to be infected. Our results suggest that the Siberian chipmunk may be an important reservoir host for LB.

Age- and sex-dependent effects of long-term zinc supplementation on essential trace element status and lipid metabolism in European subjects: the Zenith Study.

Given the key role of Zn in many physiological functions, optimal Zn status could be a predictive parameter of successful ageing. However, the benefit of Zn supplementation is still a matter of debate since Zn supplementation has been reported to be associated with the alteration of Cu status and lipid metabolism. As part of the Zenith Project, the present study aimed to investigate, in free-living healthy European middle-aged and older subjects, the effect of Zn supplementation on the biochemical status of Zn, Fe and Cu and on lipid profile. Volunteers aged 55-70 (n 188) and 70-85 (n 199) years old participated in a double-blinded, randomised study and received a daily placebo, or Zn as 15 or 30 mg for 6 months. Zn supplementation did not significantly modify erythrocyte Zn levels or erythrocyte Cu,Zn-superoxide dismutase activity. But Zn supplementation at 15 or 30 mg/d for 6 months increased significantly serum Zn levels and Zn urinary excretion with no major adverse effects on Fe and Cu status or on lipid metabolism. However, Zn supplementation at 30 mg/d showed some age- and sex-dependent alterations in Fe status or lipid profile. Therefore, with respect to the key role of an optimal Zn status in successful ageing, Zn supplementation at 15 mg/d, when necessary, could be safely proposed regarding lipids and the risk of interaction with Fe and Cu.

Association of virulence genotype with phylogenetic background in comparison to different seropathotypes of Shiga toxin-producing Escherichia coli isolates.

The distribution of virulent factors (VFs) in 287 Shiga toxin-producing Escherichia coli (STEC) strains that were classified according to Karmali et al. into five seropathotypes (M. A. Karmali, M. Mascarenhas, S. Shen, K. Ziebell, S. Johnson, R. Reid-Smith, J. Isaac-Renton, C. Clark, K. Rahn, and J. B. Kaper, J. Clin. Microbiol. 41:4930-4940, 2003) was investigated. The associations of VFs with phylogenetic background were assessed among the strains in comparison with the different seropathotypes. The phylogenetic analysis showed that STEC strains segregated mainly in phylogenetic group B1 (70%) and revealed the substantial prevalence (19%) of STEC belonging to phylogenetic group A (designated STEC-A). The presence of virulent clonal groups in seropathotypes that are associated with disease and their absence from seropathotypes that are not associated with disease support the concept of seropathotype classification. Although certain VFs (eae, stx(2-EDL933), stx(2-vha), and stx(2-vhb)) were concentrated in seropathotypes associated with disease, others (astA, HPI, stx(1c), and stx(2-NV206)) were concentrated in seropathotypes that are not associated with disease. Taken together with the observation that the STEC-A group was exclusively composed of strains lacking eae recovered from seropathotypes that are not associated with disease, the "atypical" virulence pattern suggests that STEC-A strains comprise a distinct category of STEC strains. A practical benefit of our phylogenetic analysis of STEC strains is that phylogenetic group A status appears to be highly predictive of "nonvirulent" seropathotypes.