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Key parasite transmission parameters are difficult to obtain from elusive wild animals. For Echinococcus multilocularis, the causative agent of alveolar echinococcosis (AE), the red fox is responsible for most of the environmental contamination in Europe. The identification of individual spreaders of E. multilocularis environmental contamination is crucial to improving our understanding of the ecology of parasite transmission in areas of high endemicity and optimising the effectiveness of prevention and control measures in the field. Genetic faecal sampling appears to be a feasible method to gain information about the faecal deposition of individual animals. We conducted a 4 year faecal sampling study in a village that is highly endemic for E. multilocularis, to assess the feasibility of individual identification and sexing of foxes to describe individual infection patterns. Individual fox identification from faecal samples was performed by obtaining reliable genotypes from 14 microsatellites and one sex locus, coupled with the detection of E. multilocularis DNA, first using captive foxes and then by environmental sampling. From a collection of 386 fox stools collected between 2017 and 2020, tested for the presence of E. multilocularis DNA, 180 were selected and 124 samples were successfully genotyped (68.9%). In total, 45 unique individual foxes were identified and 26 associated with at least one sample which tested positive for E. multilocularis (Em(+)). Estimation of the population size showed the fox population to be between 29 and 34 individuals for a given year and 67 individuals over 4 years. One-third of infected individuals (9/26 Em(+) foxes) deposited 2/3 of the faeces which tested positive for E. multilocularis (36/60 Em(+) stools). Genetic investigation showed a significantly higher average number of multiple stools for females than males, suggesting that the two sexes potentially defecated unequally in the studied area. Three partially overlapping clusters of fox faeces were found, with one cluster concentrating 2/3 of the total E. multilocularis-positive faeces. Based on these findings, we estimated that 12.5 million E. multilocularis eggs were produced during the study period, emphasizing the high contamination level of the environment and the risk of exposure faced by the parasite hosts., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Alveolar echinococcosis is considered to be the most serious zoonosis in the Northern Hemisphere's cold or temperate regions. In Europe, the parasite has a sylvatic life cycle based on predator-prey interactions, mainly between red foxes and small rodents. Echinococcus multilocularis has been observed to have spread across Europe over the last three decades. In France, a westward spread of the parasite's known endemic areas has been described. In this study, a retrospective analysis of fox feces by real-time PCR was carried out in four départements not previously investigated and considered free along with two endemic control departments. The fox feces collected from 2000 to 2004 were analyzed by real-time PCR. Fecal prevalence in the two endemic departments of Doubs and Côte d'Or were estimated at 12 % [6.4-20.0 %] and 4.2 % [1.1-10.3 %], respectively. No positive samples were identified among the 72 feces collected in Drôme or the 112 from Allier, which is consistent with the very low expected prevalence should the parasite be present. Three positive samples were recovered in the Seine-Maritime and Hautes-Alpes départements, resulting in a prevalence of 3.5 % [0.7-10.0 %] and 2.5 % [0.5-7.1 %], respectively. From now on, Hautes-Alpes constitutes the new southern border of the endemic areas in France and confirms the southward expansion previously highlighted. Real-time copro-PCR proved useful in identifying new endemic areas even with low prevalence. Due to the spread of E. multilocularis in France and associated zoonotic risk, it is necessary to expand surveillance in order to fully define all the country's endemic areas. On a continental scale, the development and harmonization of surveillance programs are now needed in order to obtain a global overview of the presence of E. multilocularis and to tailor potential countermeasures.

The oncosphere stage of Echinococcus multilocularis in red fox stools can lead, after ingestion, to the development of alveolar echinococcosis in the intermediate hosts, commonly small mammals and occasionally humans. Monitoring animal infection and environmental contamination is a key issue in public health surveillance. We developed a quantitative real-time PCR technique (qPCR) to detect and quantify E. multilocularis DNA released in fox faeces. A qPCR technique using a hydrolysis probe targeting part of the mitochondrial gene rrnL was assessed on (i) a reference collection of stools from 57 necropsied foxes simultaneously investigated using the segmental sedimentation and counting technique (SSCT) (29 positive for E. multilocularis worms and 28 negative animals for the parasite); (ii) a collection of 114 fox stools sampled in the field: two sets of 50 samples from contrasted endemic regions in France and 14 from an E. multilocularis-free area (Greenland). Of the negative SSCT controls, 26/28 were qPCR-negative and two were weakly positive. Of the positive SSCT foxes, 25/29 samples were found to be positive by qPCR. Of the field samples, qPCR was positive in 21/50 (42%) and 5/48 (10.4%) stools (2 samples inhibited), originating respectively from high and low endemic areas. In faeces, averages of 0.1 pg/μl of DNA in the Jura area and 0.7 pg/μl in the Saône-et-Loire area were detected. All qPCR-positive samples were confirmed by sequencing. The qPCR technique developed here allowed us to quantify environmental E. multilocularis contamination by fox faeces by studying the infectious agent directly. No previous study had performed this test in a one-step reaction., (Copyright © 2013 Elsevier B.V. All rights reserved.)

During 2005-2010, we investigated Echinococcus multilocularis infection within fox populations in a large area in France. The parasite is much more widely distributed than hitherto thought, spreading west, with a much higher prevalence than previously reported. The parasite also is present in the large conurbation of Paris.

The functional response of predators to prey density variations has previously been investigated in order to understand predation patterns. However, the consequences of functional response on parasite transmission remain largely unexplored. The rodents Microtus arvalis and Arvicola terrestris are the main prey of the red fox Vulpes vulpes in eastern France. These species are intermediate and definitive hosts of the cestode Echinococcus multilocularis. We explored the dietary and contamination responses of the red fox to variations in prey density. The dietary response differed between the two prey species: no response for M. arvalis and a type III-like (sigmoidal) response for A. terrestris that shows possible interference with M. arvalis. The fox contamination response followed a type II shape (asymptotic) for both species. We conclude that fox predation is species specific and E. multilocularis transmission is likely to be regulated by a complex combination of predation and immunologic factors. These results should provide a better understanding of the biological and ecological mechanisms involved in the transmission dynamics of trophically transmitted parasites when multiple hosts are involved. The relevance of the models of parasite transmission should be enhanced if non-linear patterns are taken into account.

Background: Alveolar echinococcosis (AE) is a severe helminth disease affecting humans, which is caused by the fox tapeworm Echinococcus multilocularis. AE represents a serious public health issue in larger regions of China, Siberia, and other regions in Asia. In Europe, a significant increase in prevalence since the 1990s is not only affecting the historically documented endemic area north of the Alps but more recently also neighbouring regions previously not known to be endemic. The genetic diversity of the parasite population and respective distribution in Europe have now been investigated in view of generating a fine-tuned map of parasite variants occurring in Europe. This approach may serve as a model to study the parasite at a worldwide level., Methodology/principal Findings: The genetic diversity of E. multilocularis was assessed based upon the tandemly repeated microsatellite marker EmsB in association with matching fox host geographical positions. Our study demonstrated a higher genetic diversity in the endemic areas north of the Alps when compared to other areas., Conclusions/significance: The study of the spatial distribution of E. multilocularis in Europe, based on 32 genetic clusters, suggests that Europe can be considered as a unique global focus of E. multilocularis, which can be schematically drawn as a central core located in Switzerland and Jura Swabe flanked by neighbouring regions where the parasite exhibits a lower genetic diversity. The transmission of the parasite into peripheral regions is governed by a "mainland-island" system. Moreover, the presence of similar genetic profiles in both zones indicated a founder event.

In order to identify the respective importance of the ecological and biological factors involved in the transmission of Echinococcus multilocularis, we estimated grassland vole intermediate host (Microtus sp. and Arvicola terrestris) population densities, in relation to the diet of the definitive host (red fox, Vulpes vulpes) and with the prevalence of E. multilocularis in the fox population. The study was conducted in the Ardennes, north-eastern France, which is an area with a high incidence of alveolar echinococcosis. Surface index methods showed that Microtus was the most abundant intermediate host in the area. Furthermore, Microtus was present in one-third of the 144 faeces and 98 stomach content samples examined and represented more than two-thirds of the rodent occurrences. Red fox predation on Microtus was significantly correlated with Microtus relative abundance. In contrast, the relative abundance of A. terrestris was very low. This species, as well as Clethrionomys glareolus and Apodemus sp., was little consumed. E. multilocularis prevalence in foxes was determined from carcasses and reached 53% (95% confidence interval 45-61%). Intensity of infection varied from 2 to 73,380 worms per fox, with 72% of the sampled worm burden harboured by 8% of the sampled foxes. The selected explanatory variables (sex, year, age class, health and nutritional condition, and season) failed to predict prevalence rate and worm burden. The high prevalence rate in foxes indicates the possibility of intense E. multilocularis transmission, apart from periods, or in landscapes, favourable to large population outbreaks of grassland rodents.

• Exposure to Echinococcus multilocularis was investigated in an endemic area. • The protocol was based on a non-invasive copro-sampling approach using qPCRs and a flotation technique. • Among 96 carnivore faeces containing Echinococcus multilocularis DNA, 32 contained E. multilocularis eggs. • Echinococcus multilocularis DNA and eggs were detected more frequently in fox faeces than cat or dog faeces. • The faecal prevalence of E. multilocularis DNA and eggs was overdispersed with the same geographical patterns. The eggs of Echinococcus multilocularis , the infectious stage, are spread into the environment through wild and domestic carnivore faeces. The spatial location of the faeces containing infective E. multilocularis eggs is a key parameter for studying areas of exposure and understanding the transmission processes to the intermediate hosts and humans. Echinococcus multilocularis faecal prevalence is often assessed by detecting E. multilocularis DNA, not necessarily eggs. This work aimed to determine the percentage of faeces containing E. multilocularis eggs in a rural town and its surroundings and whether this level of precision is relevant in assessing exposure to E. multilocularis. For this purpose, we developed a combined molecular and microscopic approach to investigate the E. multilocularis exposure of potential hosts in the environment from field-collected carnivore faeces. Carnivore defecation patterns were then spatialized to study the spatial distribution of E. multilocularis. Faeces were screened for E. multilocularis DNA using a specific real-time quantitative PCR (qPCR). Echinococcus multilocularis eggs were morphologically identified from E. multilocularis- specific qPCR-positive faeces after sucrose flotation and individually confirmed through specific PCR and sequencing. The spatial distribution of E. multilocularis was studied using Kulldorff statistics. Echinococcus multilocularis eggs were identified mostly in fox faeces positive for E. multilocularis DNA by qPCR (n = 27/70) and only from 1 of 15 copro-samples from dogs and 1 of 5 from cats. The faecal prevalence of E. multilocularis DNA and eggs was overdispersed, with the same geographical patterns. These data suggest that E. multilocularis DNA and/or egg detection in carnivore faeces, mainly that of foxes, is appropriate in ecological studies of E. multilocularis transmission. [ABSTRACT FROM AUTHOR]