Rural and urban distribution of wild and domestic carnivore stools in the context of Echinococcus multilocularis environmental exposure.

In zoonotic infections, the relationships between animals and humans lead to parasitic disease with severity that ranges from mild symptoms to life-threatening conditions. In cities and their surrounding areas, this statement is truer with the overcrowding of the protagonists of the parasites' life cycle. The present study aims to investigate the distribution of a parasite, Echinococcus multilocularis, which is the causative agent of alveolar echinococcosis, using copro-sampling in historically endemic rural settlements of the eastern part of France and in newly endemic areas including urban parks and settlements surrounding Paris. Based on 2741 morphologically identified and geolocalized copro-samples, the density of fox faeces was generally higher in the surrounding settlements, except for one rural area where the faeces were at larger density downtown in the winter. Fox faeces are rare but present in urban parks. Dog faeces are concentrated in the park entrances and in the centre of the settlements. DNA was extracted for 1530 samples that were collected and identified from fox, dog, cat, stone marten and badger carnivore hosts. Echinococcus multilocularis diagnosis and host faecal tests were performed using real-time PCR. We failed to detect the parasite in the surroundings of Paris, but the parasite was found in the foxes, dogs and cats in the rural settlements and their surroundings in the historically endemic area. A spatial structuring of the carnivore stool distribution was highlighted in the present study with high densities of carnivore stools among human occupied areas within some potentially high-risk locations.

Echinococcus multilocularis management by fox culling: An inappropriate paradigm.

With the ongoing spread of Echinococcus multilocularis in Europe, sanitary authorities are looking for the most efficient ways of reducing the risk for human populations. Fox culling is one particular tool that has recently shifted from predation control to population health management. Our study aims to assess the effectiveness of this tool in limiting E. multilocularis prevalence in fox populations in France. During four years, a culling protocol by night shooting from cars was implemented around the city of Nancy (eastern France) representing ∼1700h of night work and ∼15,000km driven. The 776 foxes killed represented an overall increase of 35% of the pressure on the fox population over 693km2. Despite this consequent effort of culling, not only did night shooting of foxes fail to decrease the fox population, but it resulted in an increase in E. multilocularis prevalence from 40% to 55% while remaining stable in an adjacent control area (585km2). Though no significant change in age structure could be described, an increase in immigration and local recruitment is the best hypothesis for population resilience. The increase in prevalence is therefore considered to be linked to a higher rate of juvenile movement within the culled area shedding highly contaminated faeces. We therefore advocate managers to consider alternative methods such as anthelmintic baiting, which has been proven to be efficient elsewhere, to fight against alveolar echinococcosis.

Hedgehogs and Mustelid Species: Major Carriers of Pathogenic Leptospira, a Survey in 28 Animal Species in France (20122015).

Human leptospirosis is a zoonotic and potentially fatal disease that has increasingly been reported in both developing and developed countries, including France. However, our understanding of the basic aspects of the epidemiology of this disease, including the source of Leptospira serogroup Australis infections in humans and domestic animals, remains incomplete. We investigated the genetic diversity of Leptospira in 28 species of wildlife other than rats using variable number tandem repeat (VNTR) and multispacer sequence typing (MST). The DNA of pathogenic Leptospira was detected in the kidney tissues of 201 individuals out of 3,738 tested individuals. A wide diversity, including 50 VNTR profiles and 8 MST profiles, was observed. Hedgehogs and mustelid species had the highest risk of being infected (logistic regression, OR = 66.8, CI95% = 30.9-144 and OR = 16.7, CI95% = 8.7-31.8, respectively). Almost all genetic profiles obtained from the hedgehogs were related to Leptospira interrogans Australis, suggesting the latter as a host-adapted bacterium, whereas mustelid species were infected by various genotypes, suggesting their interaction with Leptospira was different. By providing an inventory of the circulating strains of Leptospira and by pointing to hedgehogs as a potential reservoir of L. interrogans Australis, our study advances current knowledge on Leptospira animal carriers, and this information could serve to enhance epidemiological investigations in the future.

Retrospective analyses of fox feces by real-time PCR to identify new endemic areas of Echinococcus multilocularis in France.

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.

Echinococcus multilocularis infections in dogs from urban and peri-urban areas in France.

Echinococcus multilocularis is the causative agent of alveolar echinococcosis, a severe zoonotic disease. It is maintained through a sylvatic life cycle based on predator-prey interactions mainly between foxes and rodents. Dogs are also good definitive hosts; and due to their close proximity to humans, they may represent a major risk factor for the occurrence of human cases. In two medium-sized cities of Eastern France (Annemasse and Pontarlier), located in highly endemic areas, 817 dog feces samples were collected and analyzed by a flotation technique followed by a multiplex PCR assay. For the first time in France, we assessed the presence of E. multilocularis DNA in four dog feces samples, in which it represents an estimated prevalence of 0.5% (95% CI; 0.1% <> 1.3%). Eight other samples presented taeniid infections from three different species (Taenia crassiceps, Taenia serialis, and Taenia polyacantha). When considering both E. multilocularis and Taenia sensu lato, prevalence rose to 0.6% in Annemasse and 2.6% in Pontarlier. In this highly endemic context, proper application of the usual deworming recommendations (70% of the dogs were treated twice a year or more) failed to prevent dog infection, particularly for hunting dogs. Our results stressed the need to adapt treatment to the environmental context and to the specific activity of dogs. Further epidemiological surveys in domestic dogs and cats using this coprological approach are still needed to obtain a better overview of infection and the associated zoonotic risk.

Westward spread of Echinococcus multilocularis in foxes, France, 2005-2010.

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.

Echinococcus multilocularis in dogs from two French endemic areas: no evidence of infection but hazardous deworming practices.

The life cycle of the zoonotic parasite Echinococcus multilocularis is predominantly sylvatic, involving foxes as definitive hosts infected by predation of rodents, the intermediate hosts. The North-Eastern French departments of Meuse and Haute-Saône are highly endemic, with an estimated fox prevalence of 41% and 36% respectively. Although most of the parasites' biomass occurs in foxes, domestic dogs can also be infected, leading to a major risk of human infection due to the close proximity of dogs and owners. In the present study, dog faeces were collected after praziquantel treatment provided by veterinarians. In all, 860 faecal samples were collected throughout Meuse (n=493) and Haute-Saône (n=367). Intestinal helminth eggs were isolated from the faeces using a flotation technique and observed by microscopy. Parasite species were identified in samples positive for taeniid eggs by sequence analysis after PCR amplification. To study the factors associated with infestation, each sample was linked to a questionnaire filled in by the dog owners. Taeniid eggs were observed in seven faecal samples (0.8%) but none of them were positive for E. multilocularis. Thus, the apparent prevalence of E. multilocularis in dog populations is lower than 1.00% for Haute-Saône and lower than 0.75% for Meuse. In Haute-Saône, a high proportion of dogs observed suspected preying on rodents were not dewormed monthly. In endemic areas, these dogs must be considered at risk of transmitting E. multilocularis to humans.