Gilles Dreyfuss

The definitive and intermediate hosts of Fasciola hepatica in the natural watercress beds in central France.

Abstract. Field investigations were carried out over a two-year period in 52 natural watercress beds located in the Limousin region of central France to list the mammal and bird species that frequented these sites. This enabled detection of the definitive hosts of Fasciola hepatica and determination of the prevalence of natural infection in snails. A total of 13 mammal and five bird species were listed in these watercress beds. Adult flukes were found in Lepus capensis (39.2%), Oryctolagus cuniculus (42.0%), and Sylvilagus floridanus (25.0%). No infection with F. hepatica was noted in the five species of rodents studied. Snails infected with F. hepatica were found in 14 watercress beds. The global prevalence of natural infection was 1.1% in Lymnaea truncatula and 0.3% in L. glabra. Among the other trematode larval forms detected, the most frequent was Haplometra cylindracea (0.5%). In the Limousin region, the presence of hares and rabbits in watercress beds ensured the continuation of the F. hepatica life cycle and permitted the subsequent infection of humans when this wild watercress was eaten.

The contamination of wild watercress with Fasciola hepatica in central France depends on the ability of several lymnaeid snails to migrate upstream towards the beds

As most natural watercress beds in central France are located upstream of the permanent habitats of two lymnaeid species, Galba truncatula and Omphiscola glabra, field investigations were made from 1999 to 2004 on 67 beds to determine why the contamination of watercress with Fasciola hepatica is irregular over time in these sites, while definitive hosts, especially lagomorphs, are regularly found infected around them. Snails are able to migrate upstream in winter and spring towards the beds, and a 4-year survey demonstrated the existence of annual variation in the colonization of these sites by snails. In the 45 beds irregularly contaminated with F. hepatica over time, 37.7–62.2%, according to the year, were not populated by lymnaeids, in spite of the presence of snail populations downstream. G. truncatula was found alone in 8.8–13.3% of sites and was the first colonizing snail in 24.3–33.3% when the two lymnaeid species successively settled in these waterholes. The colonizing ability of O. glabra was more limited, as it was observed alone in 2.2% of beds and was the first colonizing snail in only 2.2–20% of them. The distances covered by these snails significantly increased with increasing migration time. After migration, a few overwintering snails (3.8% of G. truncatula and 6.8% of O. glabra) are able to colonize the beds, but their numbers decreased when the distance of migration was greater. O. glabra migrated more quickly and reached more watercress beds than G. truncatula. However, this did not influence the natural infections of snails, which were more frequent in the F1 of G. truncatula. The variability observed in the colonization of beds by snails might explain the regular or irregular contamination of wild watercress by metacercariae of F. hepatica. One of the factors which may explain this variability is the occurrence of showers in spring, so that a few snails are able to reach watercress beds after their upstream migration.

Variability of Fasciola hepatica infection in Lymnaea ovata in relation to snail population and snail age

Abstract Bimiracidial infections of Lymnaea ovata with Fasciola hepatica were performed under laboratory conditions to determine the susceptibility of snails from six French populations to trematode infection. In five populations of L. ovata the prevalence of infection in the 1-mm groups ranged between 2.7% and 43.7% at day 35 postexposure; it decreased in the 2-mm snails and was zero in larger groups. In the snails from Thenay (periodically polluted brook) the prevalence of F. hepatica infection decreased from the 1-mm group to the 8-mm group (from 23.9% to 1.0%) and was zero in the 10-mm L. ovata. The total number of cercariae shed per snail was 18.3 in the 1-mm group, increasing to 117 in the 8-mm group. The latter findings could be interpreted as a consequence of periodic pollution in the brook of Thenay; pollution might disrupt the defense system of L. ovata and facilitate the subsequent larval development of F. hepatica.

Molecular identification of Fasciola spp. (Digenea: Fasciolidae) in Egypt

A total of 134 Egyptian liver flukes were collected from different definitive hosts (cattle, sheep, and buffaloes) to identify them via the use of PCR-RFLP and sequence analysis of the first nuclear ribosomal internal transcribed spacer (ITS1). Specimens of F. hepatica from France, as well as F. gigantica from Cameroon were included in the study for comparison. PCR products of ITS1 were subjected for digestion by RsaI restriction enzyme and visualized on agarose gel. According to RFLP pattern, Egyptian flukes were allocated into two categories. The first was identical to that of French hepatica flukes to have a pattern of 360, 100, and 60 (bp) band size, whereas the second resembled to that of Cameroonian gigantica worms to have a profile of 360, 170, and 60 bp in size. Results of RFLP analysis were confirmed by sequence analysis of representative ITS1 amplicons. No hybrid forms were detected in the present study. Taken together, this study concluded that both species of Fasciola are present in Egypt, whereas the hybrid form may be not very common.

Fasciola hepatica: the infectivity of cattle-origin miracidia had increased over the past years in central France.

A retrospective study on experimental infections of Galba truncatula, originating from four populations, was carried out to determine the infectivity of Fasciola hepatica miracidia in snails either subjected to single-miracidium or bimiracidial exposures in 2006 and to compare it with results found before 1990 in other experiments using the same protocol. In single-miracidium infections, the prevalence of snail infection was significantly greater in 2006 than in experiments performed before 1980, while snail survival at day 30 postexposure showed insignificant differences. In bimiracidial infections, snail survival in two snail groups was significantly lower in 2006 than in experiments performed from 1981 to 1990, while insignificant differences were found for the other two populations of G. truncatula. Compared to results found between 1981 and 1990, the prevalence of snail infection did not significantly differ in 2006. In each population considered alone, the mean number of cercariae shed by infected snails did not significantly vary, whatever the date of experiment and the number of miracidia used for each exposure. The greater infectivity of F. hepatica miracidia towards snails might be the consequence of specific molecules such as triclabendazole used since 1990 to treat fasciolosis in cattle from central France.

Microsatellite analysis of Fasciola spp. in Egypt

Recently, the topic of diversity in Fasciola population in Egypt is controversial. The present study was performed to study the genetic diversity of isolated flukes based on microsatellites markers. Fasciola worms were collected from different hosts and geographical locations in Egypt. Control samples of Fasciola hepatica from France as well as Fasciola gigantica from Cameroon were included in the study. Collected flukes were identified morphologically and subjected for analysis using four microsatellite markers. Results of microsatellite profile (FM1 and FM2) proved that both species of Fasciola are distributed in Egypt irrespective of geographical location and host. Nevertheless, the microsatellite profile of some analyzed loci (FM2 and FM3) proved that Egyptian flukes showed more alleles compared to the reference ones. Differences of microsatellite profile in Egyptian isolates than that of corresponding reference samples indicate the remarkable diversity of these isolates. The present results highlighted the utility of microsatellite profile to discriminate between Fasciola species and to elucidate the diversity within the species. To our knowledge, this is the first time to study microsatellite polymorphism in Fasciola populations in Egypt.

Characterisation of Pseudosuccinea columella and Radix natalensis (Gastropoda: Lymnaeidae) in Egypt using shell and molecular data

ABSTRACT Pseudosuccinea columella and Radix natalensis live in the same habitat in Egypt and are important intermediate hosts of Fasciola hepatica and F. gigantica. Our study aimed to characterise both snail species using molecular analysis and shell measurements. The ranges of morphometric parameters overlapped in the two lymnaeids, indicating that they do not clearly differentiate the two species. PCR-sequence analysis of the nuclear ribosomal small subunit rRNA and the polymorphic mitochondrial cytochrome oxidase subunit 1 (CO1) genes were used to determine the genetic identity and the potential diversity of the snails. Little intrasequence variations were detected in the sequences of both gene loci, indicating the potential homogeneity of lymnaeid populations in Egypt. Generated sequences of the mitochondrial CO1 gene locus for R. natalensis showed obvious heterogeneity compared to other sequences in GenBank. Molecular characterisation of these lymnaeids might help to understand the snails’ biodiversity in a bid to control these populations and their related diseases.

Consequences of invasion by Pseudosuccinea columella on the dynamics of native lymnaeids living on the acid soils of central France

ABSTRACT Eight road ditches located on granite and colonised by the native lymnaeids Galba truncatula or Omphiscola glabra were studied for 2 years to follow the distribution and density of these species in their habitats when the invasive lymnaeid Pseudosuccinea columella was introduced. Six other ditches populated only by a native lymnaeid or by P. columella were used as controls. The same experiment was also carried out for 3 years in seven water tanks placed in semi-natural conditions. The colonisation of P. columella was more rapid in habitats with G. truncatula than in those with O. glabra. Similarly, this colonisation led to a progressive decrease in the number of native lymnaeids in these ditches and this decrease was also faster for G. truncatula than for O. glabra. A similar decrease of G. truncatula and O. glabra was also noted when P. columella was introduced into water tanks placed in semi-natural conditions. The habitats of G. truncatula or O. glabra on the acid soils of central France can be successfully colonised by P. columella, and the establishment of this invasive species may lead to a decrease of the two native lymnaeids or even the local extinction of one of these (G. truncatula).