Species or local environment, what determines the infection of rodents by Toxoplasma gondii?

Toxoplasmosis is largely present in rural areas but its spatial distribution in this environment remains poorly known. In particular, it is unclear if areas of high density of cats, the only hosts excreting Toxoplasma gondii, constitute foci of high prevalence. To improve our understanding of the spatial distribution of T. gondii in rural areas, we performed a serological survey in rodents from two villages in France. We trapped 710 rodents including commensal rats and meadow or forest voles and mice. The presence of T. gondii was examined using PCR, mice inoculation and modified agglutination test for antibodies (MAT). We conducted multivariate and discriminant analyses to identify biological, ecological or spatial variables that could explain T. gondii serology in rodents. We then used a logistic regression to assess the relative influence of each explanatory variable. Overall seroprevalence was 4.1%. Commensal-rats were more infected (12.5%) than non-commensal species (3.7%). However, the major determinant of the risk of infection was the distance to the nearest farm (OR = 0.75 for 100 m), which explained the risk in all species or non-commensal species only. We contrast the role of species characteristics and that of the local environment, and discuss the risk of environmental contamination for humans.

Agricultural landscape and spatial distribution of Toxoplasma gondii in rural environment: an agent-based model.

BACKGROUND: Predicting the spatial distribution of pathogens with an environmental stage is challenging because of the difficulty to detect them in environmental samples. Among these pathogens, the parasite Toxoplasma gondii is the causative agent of the zoonosis toxoplasmosis, which is responsible for public health issues. Oocysts of T. gondii are excreted by infected cats in the environment, where they may survive and remain infectious for intermediate hosts, specifically rodents, during months to years. The landscape structure that determines the density and distribution of cats may thus impact the spatial distribution of T. gondii. In this study, we investigated the influences of rural settings on the spatial distribution of oocysts in the soil. METHOD: We developed a spatially explicit agent based model to study how landscape structures impact on the spatial distribution of T. gondii prevalence in its rodent intermediate host as well as contamination in the environment. The rural landscape was characterized by the location of farm buildings, which provide shelters and resources for the cats. Specifically, we considered two configurations of farm buildings, i.e. inside and outside a village. Simulations of the first setting, with farm buildings inside the village, were validated using data from previous field studies. Then, simulation results of the two settings were compared to investigate the influences of the farm locations. RESULTS: Model predictions showed a steeper relationship between distance to the nearest farm and infection levels when farm buildings, and thus cats, were concentrated in the same area than when the farms were spread over the area. The relationship between distance to the village center and level of environmental contamination also differed between settings with a potential increased risk for inhabitants when farms are located inside the village. Maps of the risk of soil contaminated with oocysts were also derived from the model. CONCLUSION: The agent-based model provides a useful tool to assess the risk of contamination by T. gondii oocysts at a local scale and determine the most at risk areas. Moreover it provides a basis to investigate the spatial dynamics of pathogens with an environmental stage.

Quantitative estimation of the viability of Toxoplasma gondii oocysts in soil.

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.

Spatial distribution of Toxoplasma gondii oocysts in soil in a rural area: Influence of cats and land use.

Toxoplasma gondii is the protozoan parasite responsible for toxoplasmosis, one of the most prevalent zoonoses worldwide. T. gondii infects humans through the ingestion of meat containing bradyzoites or through soil, food or water contaminated with oocysts. Soil contamination with oocysts is increasingly recognized as a major source of infection for humans, but has rarely been quantified directly. In this study, we investigated the spatial pattern of soil contamination with T. gondii over an area of 2.25 km(2) in a rural area of eastern France. The frequency and spatial distribution of T. gondii in soil was analyzed in relation with the factors that could influence the pattern of contamination: cats' frequency and spatial distribution and land use. According to a stratified random sampling Scheme 243 soil samples were collected. The detection of T. gondii oocysts was performed using a recent sensitive method based on concentration and quantitative PCR. Sensitivity was improved by analyzing four replicates at each sampling point. T. gondii was detected in 29.2% of samples. Soil contamination decreased with increasing distance from the core areas of cat home ranges (households and farms). However, it remained high at the periphery of the study site, beyond the boundaries of the largest cat home ranges, and was not related to land use. This pattern of contamination strongly supports the role of inhabited areas which concentrate cat populations as sources of risk for oocyst-induced infection for both humans and animals. Moreover, soil contamination was not restricted to areas of high cat density suggesting a large spatial scale of environmental contamination, which could result from T. gondii oocysts dissemination through rain washing or other mechanisms.

Toxoplasmosis in natural populations of ungulates in France: prevalence and spatiotemporal variations.

Toxoplasmosis is characterized by a complex epidemiology. The risk of infection for humans depends on their contact with infective oocysts in a contaminated environment and on the amount of tissue cysts located within consumed meat. Unfortunately, the prevalence of tissue cysts is largely unknown for game species. Although herbivorous game species are a source of infection for humans, the level of infection found in wildlife can also be used to estimate environmental contamination. The aim of this study was to estimate the prevalence of Toxoplasma gondii infection and analyze its temporal dynamics in one population of chamois (Rupicapra rupicapra), one of mouflon (Ovis gmelini musimon), and two of roe deer (Capreolus capreolus) in France, surveyed during a period of 6 to 28 years. Taking into account individual risk factors, we specifically analyzed the relationship between T. gondii prevalence and meteorological conditions that may influence oocyst survival. Serum samples from 101 chamois, 143 mouflons, and 1155 roe deer were tested for antibodies against T. gondii using the modified agglutination test (MAT), an enzyme-linked immunosorbent assay (ELISA) assay, or both. Using MAT with a threshold of 1:6, seroprevalence was 14.7% in mouflon, 16.8% in chamois, and 43.7% in roe deer. In mouflon and roe deer, seroprevalence was positively correlated with age and/or body mass, in accordance with the hypothesis that antibodies have long-term persistence. In roe deer, seropositivity differed between the two populations and changed linearly over time between 1983 and 2010, increasing by a factor 1.75 every 10 years. Moreover, in this species, the highest prevalences were found during dry and cold years or during warm and moist years, depending on the population. Our results suggest that the risk for people to acquire infection through game meat increases over time, but with high variability according to the population of origin and meteorological conditions of the year.

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.