Biology subjects

Pollet, T.

Publications and source records attributed to Pollet, T..

11 recordsLinked to original sources

bioRxiv2026

Microbial communities in Ornithodoros phacochoerus ticks: spatial structure and influence of African swine fever virus infectious status

AO_SCPLOWBSTRACTC_SCPLOWOrnithodoros phacochoerus are nidicolous soft ticks of the Ornithodoros moubata complex of species known to be vectors of the African swine fever (ASF) virus. These Ornithodoros ticks depend on endosymbionts to produce essential nutrients necessary for their development. However, endosymbionts are only a part of the complex microbiota hosted by the tick. This microbiota often includes primary or secondary endosymbionts, commensal species from the environment, and, most of the time, some pathogens. The present study was performed to understand the organization and spatial distribution of the microbiota of O. phacochoerus. One of the objectives was to investigate if the pathogen of interest (ASF virus) is involved in the organization of the microbiota through pathogen-induced dysbiosis or other interactions. For this purpose, 704 O. phacochoerus ticks were collected from two conservation areas in Mozambique. Sequencing was performed targeting the V3-V4 region of the 16S rRNA gene, and the resulting dataset was processed using FROGS to characterize the bacterial microbiota hosted by the ticks. The results indicate that the microbiota of Ornithodoros phacochoerus contains very low bacterial diversity, with one primary endosymbiont (Francisella-like endosymbiont), one potential secondary endosymbiont (Rickettsiella), and very few environmental or pathogenic bacterial species. We found a clear spatial structure of the microbiota, with ticks from the same sampling site showing similar patterns. On the contrary, no association with the infectious status for African swine fever virus was detected, suggesting that this pathogen does not shape Ornithodoros microbial communities. Our results on tick - microbiota - pathogen - environment interactions in nidicolous soft ticks, showed patterns that differ from most hard tick studies.

microbiology↗

bioRxiv2025

Population genetics of the nidicolous soft tick Ornithodoros phacochoerus in the context of African swine fever sylvatic cycle

Ticks of the species Ornithodoros phacochoerus are endophilic soft ticks which infest warthog burrows. Like other Ornithodoros of the O. moubata species complex, O. phacochoerus is a vector of the African swine fever virus and participates in the maintenance of the virus among warthogs in the sylvatic cycle in Southern and Eastern African countries. In this study, the population genetic structure of O. phacochoerus was investigated using sixteen microsatellite markers. After sampling campaigns that took place between 2020 and 2022, we analyzed 684 ticks from 21 warthog burrows and resting sites from Coutada 9 Game Reserve, Macossa District, Mozambique, and 138 ticks from 3 warthog burrows and resting sites from Gorongosa National Park, Gorongosa District, Mozambique. Burrows and resting sites are regularly used by the common warthog Phacochoerus africanus which is thought to be responsible for the movements of O. phacochoerus between sites. After genotyping, the observed genetic variation followed a model of isolation by distance with a structure at the level of the sampling sites (burrows and resting sites). Gorongosa National Park and Coutada 9 Game Reserve appeared to be completely isolated from each other in terms of gene flow, and Coutada 9 Game Reserve showed a clear signature of a bottleneck effect. Effective population sizes within sampling sites were quite low (4.1 individuals), with an estimated migration rate of 35% and a mean dispersal distance of 209 meters per tick generation. These results suggested frequent tick movements between burrows at a small geographic scale, due to warthog movements between the burrows. As both wildlife conservation areas are positive for African swine fever virus, these results reinforce the suspected role of the ticks in the sylvatic cycle of the virus, as infected ticks could be moved from one burrow to another, maintaining the presence of the virus in several sites of the conservation areas. This confirms the importance of maintaining buffer zones around conservation areas, buffer zones that should remain free of any domestic pigs to prevent vector spillover from the sylvatic cycle to the domestic cycle.

ecology↗

bioRxiv2025

Contrasting temporal patterns and associations in Hyalomma marginatum microbial communities: key insights for the development of novel tick and tick-borne diseases control tools

BackgroundHyalomma marginatum is an invasive tick species in southern mainland France that can carry several pathogens of human and animal interest. Because the tick microbiota represents a major factor that can potentially modulate the pathogen acquisition and transmission and might become a new control tool against ticks and tick-borne diseases, it is more than essential to identify the composition of the H. marginatum microbiota, its temporal dynamics and interactions (statistical association) between members of the tick microbiota. MethodsFrom monthly tick samplings performed in the same site between February to September 2022, 281 adult ticks H. marginatum were collected from horses. The microbiota composition was characterised using a high throughput sequencing approach. Different statistical analyses allowed us to assess the influence of several factors (month, season, tick sex) on the H. marginatum microbial communities and reveal potential interactions among members of these communities. ResultsApart of known obligate endosymbionts Francisella and Midichloria, and the hypothesised facultative endosymbiont (Rickettsia) that dominated the microbiota of H. marginatum, we detected Staphylococcus, Corynebacterium, Williamsia and Mycobacterium, usually described as commensal and/or environmental bacteria. The microbiota composition and bacterial networks differed between males and females, with males being more diverse and composed of more environmental bacteria. We reported several temporal shifts for both males and females into the microbiota composition and bacterial networks. The temporal shifts observed for females were more chaotic in terms of movements among nodes, compared to the male microbial communities that exhibited a more organised and stable dynamics. ConclusionsThe influence of tick sex and time on the holobiont H. marginatum underlines the importance of the scale at which the study is conducted. HighlightsO_LIObligate endosymbionts dominate the Hyalomma marginatum microbiota C_LIO_LIEnvironnemental bacteria are abundant in Hyalomma marginatum microbiota C_LIO_LIBoth the bacterial composition and associations differed according to the tick sex C_LIO_LIBoth the bacterial composition and associations were marked by many temporal shifts for both males and females C_LIO_LIKeystone taxa in tick microbiota were highly variable from one month to another C_LI

microbiology↗

bioRxiv2024

Temporal variability of the Hyalomma marginatum-borne pathogens in a sentinel site of the Occitanie region (France), a focus on the intriguing dynamics of Rickettsia aeschlimannii

Spatio-temporal scales have a clear influence on microbial community distribution and diversity and are thus essential to consider to identify and study the dynamics of microorganisms. The invasive tick species Hyalomma marginatum has recently become established in southern France. It carries pathogens of medical and veterinary interest including the Crimean-Congo hemorrhagic fever virus, Rickettsia aeschlimannii, Theileria equi and Anaplasma phagocytophilum among others. While the pathogenic communities of H. marginatum were identified and their spatial distribution characterised, their temporal dynamics are still unknown. We performed a monthly H. marginatum tick collection from February to September 2022 in a sentinel site in southern France in order to study their presence and temporal dynamics. On the 281 ticks analysed, we detected pathogens included R. aeschlimannii, A. phagocytophilum and T. equi with infection rates reaching 47.0%, 4.6% and 11.0% respectively. Overall, 14.6% of ticks were infected with at least Theileria or Anaplasma, with monthly fluctuations ranging from 2.9% to 28.6%. Strong temporal patterns were observed for each of the detected pathogens, particularly for R. aeschlimannii whose infection rates drastically increased at the beginning of summer, correlated with the monthly mean temperatures in the sentinel site. Based on these results, we hypothesized that R. aeschlimannii might be a secondary symbiont of H. marginatum that could play a role into stress response to temperature. The analysis of monthly and seasonal fluctuations in H. marginatum-borne pathogens allowed us to conclude that the risk of infection is present throughout H. marginatum activity period, but particularly in summer. HighlightsO_LIStrong monthly fluctuations of pathogen infection rates were observed especially for Rickettsia aeschlimannii, currently identified as a human pathogen whose pathogenic status in humans and its symbiotic status in H. marginatum are both questioned. C_LIO_LIThe increase in temperature is correlated with the increase in Rickettsia aeschlimannii infection rates, providing clues about its potential function as a symbiont in H. marginatum. C_LIO_LITicks are recurrently infected with at least one other pathogen belonging to Theileria or Anaplasma genera with monthly fluctuations ranging from 2.9% to 28.6%. C_LIO_LIThe monthly dynamics of H. marginatum-borne pathogens are important to consider to assess the risk posed by this tick. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=81 SRC="FIGDIR/small/589770v1_ufig1.gif" ALT="Figure 1"> View larger version (24K): org.highwire.dtl.DTLVardef@b703d5org.highwire.dtl.DTLVardef@15cc273org.highwire.dtl.DTLVardef@e238d4org.highwire.dtl.DTLVardef@ebe1_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

bioRxiv2024

Spatial patterns of Hyalomma marginatum-borne pathogens in the Occitanie region (France), a focus on the intriguing dynamics of Rickettsia aeschlimannii

Hyalomma marginatum is an invasive tick species recently established in mainland southern France. This tick is known to host a diverse range of human and animal pathogens such as Rickettsia aeschlimannii, Theileria equi, Anaplasma phagocytophilum, Anaplasma marginale, Ehrlichia minasensis, Crimean-Congo hemorrhagic fever and West Nile virus. While information about the dynamics of these pathogens is crucial to assess disease risk and develop effective monitoring strategies, few data on the spatial dynamics of these pathogens are currently available. We thus collected ticks in 27 sites in the Occitanie region to characterize spatial patterns of H. marginatum- borne pathogens. Several pathogens have been detected: Theileria equi (9.2%), Theileria orientalis (0.2%), Anaplasma phagocytophilum (1.6%), Anaplasma marginale (0.8%) and Rickettsia aeschlimannii (87.3%). Interestingly, we found a spatial clustered distribution for the pathogen R. aeschlimannii between two geographically isolated areas with infection rates and bacterial loads significantly lower in Herault/Gard departments (infection rate 78.6% in average) compared to Aude/Pyrenees-Orientales departments (infection rate 92.3% in average). At a smaller scale, R. aeschlimannii infection rates varied from one site to another, ranging from 29% to 100%. Overall, such high infection rates (87.3% in average) and the effective maternal transmission of R. aeschlimannii might suggest a role as a tick symbiont in H. marginatum. Moreover, currently identified as a human pathogen, such results also question about its pathogenic status in humans given the low number of human cases. Further studies are thus needed to understand both the status and the role of R. aeschlimannii in H. marginatum ticks. IMPORTANCETicks are obligatory hematophagous arthropods which transmit pathogens of medical and veterinary importance. Their infections cause serious health issues in humans and considerable economic loss in domestic animals. Information about the presence of pathogens in ticks and their dynamics is crucial to assess disease risk for public and animal health. Analysing tick-borne pathogens in ticks collected in 27 sites in the regions Occitanie, our results highlight clear spatial patterns in the Hyalomma marginatum-borne pathogen distribution and strengthen the postulate that it is essential to develop effective monitoring strategies and consider the spatial scale to better characterize the circulation of tick-borne pathogens.

microbiology↗

bioRxiv2024

Tick-borne pathogens identified in Hyalomma marginatum (Acari: Ixodidae) collected from various vertebrate hosts in the South of France

Hyalomma marginatum is a common ectoparasitic tick of ungulates, lagomorphs, insectivores, ground-foraging birds, observed in Corsica for decades, but whose permanent establishment in mainland France is very recent. This species is known to be one of the main vectors of the Crimean-Congo Hemorrhagic Fever virus, but also of various parasitic, bacterial or viral pathogens. In this study, we investigated the molecular infection rates of numerous tick-borne pathogens in H. marginatum ticks mainly sampled on horses, and occasionally on other animal species, from the French Mediterranean rim and Corsica between 2016 and 2020. In total, 1, 195 DNA and RNA purified from individual ticks or pools of ticks were screened for 26 microbial genera or species (viruses, bacteria and parasites), using a high-throughput microfluidic real-time PCR system (BioMark dynamic array system, Standard Biotools). For individual ticks and pooled ones, respectively, the most prevalent tick-borne microorganisms were Francisella-like endosymbionts at 97.0% and 96.8%, followed by Rickettsia aeschlimannii (76.4% and 96.4%), Theileria spp. and Theileria equi (3,5% and 0%; 1,9% and 5,8%), Anaplasma phagocytophilum (3.7% and 6.7%), and West Nile virus (0.1% and 0.4%). Babesia occultans (0.9%), Ehrlichia minasensis (0.3%), and Coxiella-like endosymbionts (0.1%) were only detected in individual ticks. Our study provides an overview of the diversity of microorganisms and tick-borne pathogens detected in the invasive tick H. marginatum in Corsica and the continental departments of the Mediterranean rim. Our study opens up new research perspectives on the epidemiology of tick-borne pathogens carried by H. marginatum and on the associated public and veterinary health risks.

microbiology↗

bioRxiv2023

New insights into the role of the microbiota in Ornithodoros moubata: Influence of endosymbionts on the reproductive fitness of the tick

AO_SCPLOWBSTRACTC_SCPLOWOver the past decade, many studies have demonstrated the crucial role of the tick microbiome in tick biology. The soft tick Ornithodoros moubata is a hematophagous ectoparasite of Suidae, best known for transmitting the African swine fever virus. Its bacterial microbiota is characterized by a high prevalence of Francisella-like and Rickettsia endosymbionts. The present study aims to better understand the potential influence of these two major members of the tick microbiota on the reproductive fitness of O. moubata. A total of 132 adult female ticks were treated with antibiotics using gentamycin or rifampicin added to the blood meal. Half of the ticks were also supplemented with B vitamins to address the nutritional role of endosymbionts. Over two periods of 50 days, several traits related to the reproductive fitness were monitored to investigate the importance of Francisella and Rickettsia for these traits. It appeared that most of the reproductive parameters considered were not affected. However, antibiotic treatments induced an increase in the tick survival, indicating a potential fitness cost of harboring endosymbionts during the tick reproductive cycle. Similarly, 366 first stage nymphs of Ornithodoros moubata were exposed to the same treatments for molecular quantification of both endosymbionts. Results from qPCR suggested that the treatments had a bacteriostatic effect on endosymbionts without completely eliminating neither Francisella-like endosymbiont nor Rickettsia.

ecology↗

bioRxiv2020

Temporal patterns in Ixodes ricinus microbial communities: an insight into tick-borne microbe interactions

BackgroundTicks transmit pathogens of medical and veterinary importance, and represent an increasing threat for human and animal health. Important steps in assessing disease risk and developing possible new future control strategies involve identifying tick-borne microbes, their temporal dynamics and interactions. MethodsUsing high throughput sequencing, we studied the microbiota dynamics of Ixodes ricinus from 371 nymphs collected monthly over three consecutive years in a peri-urban forest. After adjusting a Poisson Log Normal model to our data set, the implementation of a principal component analysis as well as sparse network reconstruction and differential analysis allowed us to assess inter-annual, seasonal and monthly variability of I. ricinus microbial communities as well as their interactions. ResultsAround 75% of the detected sequences belonged to five genera known to be maternally inherited bacteria in arthropods and potentially circulating in ticks: Candidatus Midichloria, Rickettsia, Spiroplasma, Arsenophonus and Wolbachia. The structure of the I. ricinus microbiota was temporally variable with interannual recurrence and seemed to be mainly driven by OTUs belonging to environmental genera. The total network analysis revealed a majority of positive (partial) correlations. We identified strong relationships between OTUs belonging to Wolbachia and Arsenophonus, betraying the presence of the parasitoid wasp Ixodiphagus hookeri in ticks, and the well known arthropod symbiont Spiroplasma, previously documented to be involved in the defense against parasitoid wasp in Drosophila melanogaster. Other associations were observed between the tick symbiont Candidatus Midichloria and pathogens belonging to Rickettsia, probably Rickettsia helvetica. More specific network analysis finally suggested that the presence of pathogens belonging to genera Borrelia, Anaplasma and Rickettsia might disrupt microbial interactions in I. ricinus. ConclusionsHere, we identified the I. ricinus microbiota and documented for the first time the existence and recurrence of marked temporal shifts in the tick microbial community dynamics. We statistically showed strong relationships between the presence of some pathogens and the structure of the I. ricinus non-pathogenic microbes. We interestingly detected close links between some tick symbionts and the potential presence of either pathogenic Rickettsia or a parasitoid in ticks. All these new findings might be very promising for the future development of new control strategies of ticks and tick-borne diseases.

microbiology↗

bioRxiv2019

Taxon appearance from extraction and amplification steps demonstrates the value of multiple controls in tick microbiome analysis

BackgroundThe development of high throughput sequencing technologies has substantially improved analysis of bacterial community diversity, composition, and functions. Over the last decade, high throughput sequencing has been used extensively to identify the diversity and composition of tick microbial communities. However, a growing number of studies are warning about the impact of contamination brought along the different steps of the analytical process, from DNA extraction to amplification. In low biomass samples, e.g. individual tick samples, these contaminants may represent a large part of the obtained sequences, and thus generate considerable errors in downstream analyses and in the interpretation of results. Most studies of tick microbiota either do not mention the inclusion of controls during the DNA extraction or amplification steps, or consider the lack of an electrophoresis signal as an absence of contamination. In this context, we aimed to assess the proportion of contaminant sequences resulting from these steps. We analyzed the microbiota of individual Ixodes ricinus ticks by including several categories of controls throughout the analytical process: crushing, DNA extraction, and DNA amplification.\n\nResultsControls yielded a significant number of sequences (1,126 to 13,198 mean sequences, depending on the control category). Some operational taxonomic units (OTUs) detected in these controls belong to genera reported in previous tick microbiota studies. In this study, these OTUs accounted for 50.9% of the total number of sequences in our samples, and were considered contaminants. Contamination levels (i.e. the percentage of sequences belonging to OTUs identified as contaminants) varied with tick stage and gender: 76.3% of nymphs and 75% of males demonstrated contamination over 50%, while most females (65.7%) had rates lower than 20%. Contamination mainly corresponded to OTUs detected in crushing and DNA extraction controls, highlighting the importance of carefully controlling these steps.\n\nConclusionHere, we showed that contaminant OTUs from extraction and amplification steps can represent more than half the total sequence yield in sequencing runs, and lead to unreliable results when characterizing tick microbial communities. We thus strongly advise the routine use of negative controls in tick microbiota studies, and more generally in studies involving low biomass samples.

microbiology↗

bioRxiv2019

A three-years assessment of Ixodes ricinus-borne pathogens in a French peri-urban forest

BackgroundIxodes ricinus is the predominant tick species in Europe and the primary pathogen vector for both humans and animals. These ticks are frequently involved in the transmission of Borrelia burgdorferi sensu lato, the causative agents of Lyme borreliosis. While much more is known about I. ricinus tick-borne pathogen composition, information about temporal tick-borne pathogen patterns remain scarce. These data are crucial for predicting seasonal/annual patterns which could improve understanding and prevent tick-borne diseases. MethodsWe examined tick-borne pathogen (TBPs) dynamics in I. ricinus collected monthly in a peri-urban forest over three consecutive years. In total, 998 nymphs were screened for 31 pathogenic species using high-throughput microfluidic real-time PCR. ResultsWe detected DNA from Anaplasma phagocytophilum (5.3%), Rickettsia helvetica (4.5%), Borrelia burgdorferi s.l. (3.7%), Borrelia miyamotoi (1.2%), Babesia venatorum (1.5%) and Rickettsia felis (0.1%). Among all analysed ticks, 15.9% were infected by at least one of these microorganisms, and 1.3% were co-infected. Co-infections with B. afzeli/B. garinii and B. garinii/B. spielmanii were significantly over-represented. Moreover, significant variations in seasonal and/or inter-annual prevalence were observed for several pathogens (R. helvetica, B. burgdorferi s.l., B. miyamotoi, and A. phagocytophilum). ConclusionsAnalysing TBPs prevalence in monthly sampled tick over three years allowed us to assess seasonal and inter-annual fluctuations of the prevalence of TBPs known to circulate in the sampled area, but also to punctually detect less common species. All these data emphasize that sporadic tick samplings are not sufficient to determine TBPs prevalence and that regular monitoring is necessary.

epidemiology↗

bioRxiv2018

Tick-borne pathogen detection in midgut and salivary glands of adult Ixodes ricinus

BackgroundTick midgut and salivary glands represent the primary organs for pathogen acquisition and transmission, respectively. Specifically, the midgut is the first organ to have contact with pathogens during the blood meal uptake, while salivary glands along with their secretions play a crucial role in pathogen transmission to the host. Currently there is little data about pathogen composition and prevalences in I. ricinus midgut and salivary glands. The present study investigated the presence of 32 pathogen species in the midgut and salivary glands of unfed I. ricinus males and females using high-throughput microfluidic real-time PCR. Such an approach is important for enriching the knowledge about pathogen distribution in distinct tick organs which should lead to a better understanding I. ricinus-borne disease epidemiology.\n\nResultsBorrelia lusitaniae, Borrelia spielmanii, and Borrelia garinii, were detected in both midgut and salivary glands suggesting that the migration of these pathogens between these two organs might not be triggered by the blood meal. In contrast, Borrelia afzelii was detected only in the tick midgut. Anaplasma phagocytophilum and Rickettsia helvetica were the most frequently detected in ticks and were found in both males and females in the midgut and salivary glands. In contrast, Rickettsia felis was only detected in salivary glands. Finally Borrelia miyamotoi and Babesia venatorum were detected only in males in both midgut and salivary glands. Among all collected ticks, between 10 and 21% of organs were co-infected. The most common bacterial co-infections in male and female midgut and salivary glands was R. helvetica/A. phagocytophilum and R. helvetica/B. lusitaniae respectively.\n\nConclusionsAnalysing tick-borne pathogen (TBP) presence in specific tick organs enabled us to (i) highlight contrasting results with well-established transmission mechanism postulates, (ii) venture new hypotheses concerning pathogen location and migration from midgut to salivary glands, and (iii) suggest other potential associations between pathogens not previously detected at the scale of the whole tick. This work highlights the importance of considering all tick scales (i.e. whole ticks vs organs) to study TBP ecology and represents another step towards improved understanding of TBP transmission.

epidemiology↗