Marie-Lazarine Poulle

Spatial distribution of soil contamination by Toxoplasma gondii in relation to cat defecation behaviour in an urban area

In urban areas, there may be a high local risk of zoonosis due to high densities of stray cat populations. In this study, soil contamination by oocysts of Toxoplasma gondii was searched for, and its spatial distribution was analysed in relation to defecation behaviour of cats living in a high-density population present in one area of Lyon (France). Sixteen defecation sites were first identified. Cats were then repeatedly fed with marked food and the marked faeces were searched for in the defecation sites. Of 260 markers, 72 were recovered from 24 different cats. Defecation sites were frequented by up to 15 individuals. Soil samples were also examined in order to detect the presence of T. gondii using real-time PCR. The entire study area was then sampled according to cat density and vegetation cover type. Only three of 55 samples were positive and all came from defecation sites. In a second series of observations, 16 defecation sites were sampled. Eight of 62 samples tested positive, originating in five defecation sites. Laboratory experiments using experimental seeding of soil showed that the inoculated dose that can be detected in 50% of assays equals 100-1000oocysts/g, depending on the strain. This study shows that high concentrations of oocysts can be detected in soil samples using molecular methods and suggests that spatial distribution of contamination areas is highly heterogeneous. Positive samples were only found in some of the defecation sites, signifying that at-risk points for human and animal infection may be very localised.

Landscape genetics highlights the role of bank vole metapopulation dynamics in the epidemiology of Puumala hantavirus

Rodent host dynamics and dispersal are thought to be critical for hantavirus epidemiology as they determine pathogen persistence and transmission within and between host populations. We used landscape genetics to investigate how the population dynamics of the bank vole Myodes glareolus, the host of Puumala hantavirus (PUUV), vary with forest fragmentation and influence PUUV epidemiology. We sampled vole populations within the Ardennes, a French PUUV endemic area. We inferred demographic features such as population size, isolation and migration with regard to landscape configuration. We next analysed the influence of M. glareolus population dynamics on PUUV spatial distribution. Our results revealed that the global metapopulation dynamics of bank voles were strongly shaped by landscape features, including suitable patch size and connectivity. Large effective size in forest might therefore contribute to the higher observed levels of PUUV prevalence. By contrast, populations from hedge networks highly suffered from genetic drift and appeared strongly isolated from all other populations. This might result in high probabilities of local extinction for both M. glareolus and PUUV. Besides, we detected signatures of asymmetric bank vole migration from forests to hedges. These movements were likely to sustain PUUV in fragmented landscapes. In conclusion, our study provided arguments in favour of source‐sink dynamics shaping PUUV persistence and spread in heterogeneous, Western European temperate landscapes. It illustrated the potential contribution of landscape genetics to the understanding of the epidemiological processes occurring at this local scale.

Genetic characterization of the human relapsing fever spirochete Borrelia miyamotoi in vectors and animal reservoirs of Lyme disease spirochetes in France.

BackgroundIn France as elsewhere in Europe the most prevalent TBD in humans is Lyme borreliosis, caused by different bacterial species belonging to Borrelia burgdorferi sensu lato complex and transmitted by the most important tick species in France, Ixodes ricinus. However, the diagnosis of Lyme disease is not always confirmed and unexplained syndromes occurring after tick bites have become an important issue. Recently, B. miyamotoi belonging to the relapsing fever group and transmitted by the same Ixodes species has been involved in human disease in Russia, the USA and the Netherlands. In the present study, we investigate the presence of B. miyamotoi along with other Lyme Borreliosis spirochetes, in ticks and possible animal reservoirs collected in France.MethodsWe analyzed 268 ticks (Ixodes ricinus) and 72 bank voles (Myodes glareolus) collected and trapped in France for the presence of DNA from B. miyamotoi as well as from Lyme spirochetes using q-PCR and specific primers and probes. We then compared the French genotypes with those found in other European countries.ResultsWe found that 3% of ticks and 5.55% of bank voles were found infected by the same B. miyamotoi genotype, while co-infection with other Lyme spirochetes (B. garinii) was identified in 12% of B. miyamotoi infected ticks. Sequencing showed that ticks and rodents carried the same genotype as those recently characterized in a sick person in the Netherlands.ConclusionsThe genotype of B. miyamotoi circulating in ticks and bank voles in France is identical to those already described in ticks from Western Europe and to the genotype isolated from a sick person in The Netherlands. This results suggests that even though no human cases have been reported in France, surveillance has to be improved. Moreover, we showed that ticks could simultaneously carry B. miyamotoi and Lyme disease spirochetes, increasing the problem of co-infection in humans.

Quantitative Estimation of the Viability of Toxoplasma gondii Oocysts in Soil

ABSTRACT Toxoplasma gondii oocysts spread in the environment are an important source of toxoplasmosis for humans and animal species. Although the life expectancy of oocysts has been studied through the infectivity of inoculated soil samples, the survival dynamics of oocysts in the environment are poorly documented. The aim of this study was to quantify oocyst viability in soil over time under two rain conditions. Oocysts were placed in 54 sentinel chambers containing soil and 18 sealed water tubes, all settled in two containers filled with soil. Containers were watered to simulate rain levels of arid and wet climates and kept at stable temperature for 21.5 months. At nine sampling dates during this period, we sampled six chambers and two water tubes. Three methods were used to measure oocyst viability: microscopic counting, quantitative PCR (qPCR), and mouse inoculation. In parallel, oocysts were kept refrigerated during the same period to analyze their detectability over time. Microscopic counting, qPCR, and mouse inoculation all showed decreasing values over time and highly significant differences between the decreases under dry and damp conditions. The proportion of oocysts surviving after 100 days was estimated to be 7.4% (95% confidence interval [95% CI] = 5.1, 10.8) under dry conditions and 43.7% (5% CI = 35.6, 53.5) under damp conditions. The detectability of oocysts by qPCR over time decreased by 0.5 cycle threshold per 100 days. Finally, a strong correlation between qPCR results and the dose infecting 50% of mice was found; thus, qPCR results may be used as an estimate of the infectivity of soil samples.

Transmission dynamics of Toxoplasma gondii along an urban-rural gradient.

Recently, several authors have proposed that the availability of intermediate hosts (IHs) for definitive hosts (DHs) may contribute to determining the dynamics and evolutionary ecology of parasites with facultative complex life cycles. The protozoa Toxoplasma gondii may be transmitted to DHs either via predation of infected IHs through a complex life cycle (CLC) or directly from a contaminated environment through a simple life cycle (SLC). This parasite is also present in contrasting host density environments. We tested the hypothesis that the relative contributions of the CLC and SLC along an urban-rural gradient depend on the IH supply. We built and analysed a deterministic model of the T. gondii transmission cycle. The SLC relative contribution is important only in urban-type environments, i.e., with low predation rate on IHs. In contrast, the parasite is predominantly transmitted through a CLC in suburban and rural environments. The association of the two cycles enables the parasite to spread in situations of low IH availability and low DH population size for which each cycle alone is insufficient.

Environmental determinants of spatial and temporal variations in the transmission of Toxoplasma gondii in its definitive hosts

Graphical abstract

Concomitant influence of helminth infection and landscape on the distribution of Puumala hantavirus in its reservoir, Myodes glareolus

BackgroundPuumala virus, the agent of nephropathia epidemica (NE), is the most prevalent hantavirus in Europe. The risk for human infection seems to be strongly correlated with the prevalence of Puumala virus (PUUV) in populations of its reservoir host species, the bank vole Myodes glareolus. In humans, the infection risks of major viral diseases are affected by the presence of helminth infections. We therefore proposed to analyse the influence of both helminth community and landscape on the prevalence of PUUV among bank vole populations in the Ardennes, a PUUV endemic area in France.ResultsAmong the 313 voles analysed, 37 had anti-PUUV antibodies. Twelve gastro-intestinal helminth species were recorded among all voles sampled. We showed that PUUV seroprevalence strongly increased with age or sexual maturity, especially in the northern forests (massif des Ardennes). The helminth community structure significantly differed between this part and the woods or hedgerows of the southern cretes pre-ardennaises. Using PUUV RNA quantification, we identified significant coinfections between PUUV and gastro-intestinal helminths in the northern forests only. More specifically, PUUV infection was positively associated with the presence of Heligmosomum mixtum, and in a lesser extent, Aonchotheca muris-sylvatici. The viral load of PUUV infected individuals tended to be higher in voles coinfected with H. mixtum. It was significantly lower in voles coinfected with A. muris-sylvatici, reflecting the influence of age on these latter infections.ConclusionsThis is the first study to emphasize hantavirus - helminth coinfections in natural populations. It also highlights the importance to consider landscape when searching for such associations. We have shown that landscape characteristics strongly influence helminth community structure as well as PUUV distribution. False associations might therefore be evidenced if geographic patterns of helminths or PUUV repartition are not previously identified. Moreover, our work revealed that interactions between helminths and landscape enhance/deplete the occurrence of coinfections between PUUV and H. mixtum or A. muris-sylvatici. Further experimental analyses and long-term individual surveys are now required to confirm these correlative results, and to ascertain the causal links between helminth and PUUV infection risks.

Development of a sensitive method for Toxoplasma gondii oocyst extraction in soil.

Toxoplasmosis is a world-wide infection caused by Toxoplasma gondii. Oocysts disseminated in the environment by infected cats provide a major source of infection for humans and intermediate hosts. The level of soil contamination and the dynamics of this contamination are mostly unknown due to the lack of sensitivity of detection method. Our aim was to improve the detection of T. gondii oocysts in soil samples by comparing three extraction protocols (A, B and C) on unsporulated and sporulated oocysts of different strains and ages, and by testing the effect of sporulation and soil characteristics on oocyst recovery using the most efficient method. The oocyst recovery obtained using protocol C, in which the flotation solution was placed under the sample solution after the dispersion step, was at least ten-fold higher than protocols A and B, in which the sample was just filtered before flotation. The efficiency of protocol C, tested on five artificial soil matrices and four natural soils inoculated with oocysts, was lowest in soils with high proportions of sand. We recommend the protocol C for field investigations, and we advise that results should be interpreted with caution, considering the effect of soil characteristics, especially sand content, on oocyst recovery.

Effect of sex and female reproductive status on seasonal home range size and stability in rural red foxes (Vulpes vulpes)

ABSTRACT Intra-population variations in home range size can be considerable among carnivores, especially in the red fox (Vulpes vulpes). In this species, variations are not necessarily sex or age dependent. Our aim was to test whether differences in home range size and stability could be related to red fox female reproductive status, considering that red fox populations exhibit variable proportions of barren vixens. We examined differences in size and stability of seasonal home ranges of 16 foxes (three males and 13 females) radio-tracked from June 2001 to June 2003, in the Ardennes, France. Mean individual home range sizes varied widely, from 31 to 311 ha, with an average of 109 ha. The smaller a home range was, the more stable it was. Home range size was not sex dependent, but female home ranges were more stable than male ones. Reproductive females, mostly 2 y old at least, had smaller home ranges (mean: 50 ha) than non-reproductive females (mean: 174 ha), all 1 y of age. However, there was no difference in female home range stability. We hypothesized that reproductive females monopolize the richest food patches at the expense of non-reproductive females, which are younger and assumed to be subordinate and/or less experienced. Therefore, home range size correlated to food richness may be an important factor for female reproductive success. Competition for food and space could thus explain female reproductive suppression.

Performance and Accuracy of Lightweight and Low-Cost GPS Data Loggers According to Antenna Positions, Fix Intervals, Habitats and Animal Movements.

Recently developed low-cost Global Positioning System (GPS) data loggers are promising tools for wildlife research because of their affordability for low-budget projects and ability to simultaneously track a greater number of individuals compared with expensive built-in wildlife GPS. However, the reliability of these devices must be carefully examined because they were not developed to track wildlife. This study aimed to assess the performance and accuracy of commercially available GPS data loggers for the first time using the same methods applied to test built-in wildlife GPS. The effects of antenna position, fix interval and habitat on the fix-success rate (FSR) and location error (LE) of CatLog data loggers were investigated in stationary tests, whereas the effects of animal movements on these errors were investigated in motion tests. The units operated well and presented consistent performance and accuracy over time in stationary tests, and the FSR was good for all antenna positions and fix intervals. However, the LE was affected by the GPS antenna and fix interval. Furthermore, completely or partially obstructed habitats reduced the FSR by up to 80% in households and increased the LE. Movement across habitats had no effect on the FSR, whereas forest habitat influenced the LE. Finally, the mean FSR (0.90 ± 0.26) and LE (15.4 ± 10.1 m) values from low-cost GPS data loggers were comparable to those of built-in wildlife GPS collars (71.6% of fixes with LE < 10 m for motion tests), thus confirming their suitability for use in wildlife studies.