Changes in the Populations of Two Lymnaeidae and Their Infection by Fasciola hepatica and/or Calicophoron daubneyi over the Past 30 Years in Central France.

Field investigations were carried out during three periods (from 1976 to 1997, in 2013-2014, and in 2020-2021) on 39 cattle-raising farms on acidic soils to track changes in the populations of two Lymnaeidae (Galba truncatula and Omphiscola glabra) and their infection with Fasciola hepatica and/or Calicophoron daubneyi. Compared to the survey between 1976 and 1997 on these farms, there was a significant decrease in the number of the two lymnaeid populations and the size of the G. truncatula populations in both 2013-2014 and 2020-2021. This decline was significantly faster in the last nine years than it was before 2013. The area of habitats colonized by G. truncatula showed no significant variation over the years, while that of habitats with O. glabra significantly decreased in the period covered by the three surveys. The prevalence of F. hepatica infection in snails significantly decreased over the years, while C. daubneyi infection increased over time in both lymnaeid species. These changes are due to the use of triclabendazole to treat fasciolosis in ruminants since the 1990s, and are probably a consequence of the successive heatwaves that have occurred since 2018 in the region.

Fasciola hepatica: the dispersal of cercariae shed by the snail Galba truncatula.

Field investigations in 14 wild watercress beds located in the French region of Limousin, a known endemic area for distomatosis, were performed for three years to determine the distance that cercariae of Fasciola hepatica can reach in water before their encystment on the host plant. Each bed was located on the course of an open drainage furrow, while snails (Galba truncatula) lived upstream around the emergence of a source. Five plant species were collected in early April and examined to find metacercariae. Most cysts were noted on Nasturtium officinale (188 on 48.7 kg of dripped plants), followed by Helosciadium nodiflorum (125 on 33.4 kg). On the other plant species, there were few larvae. Most cercariae encysted on the plants growing in the most upstream part of each bed, usually on the first 50 cm in length. When water in the beds was fast running, the distribution of metacercariae was more limited and their number was fewer than those in the beds fed by a slow flow of water. Cercariae were able to swim or were carried away by the current up to a mean of 5 m in slow-flow waters before encysting; this distance was only 4 m in faster waters. Plants growing on the most upstream section of a watercress bed located in a drainage furrow are the most used by cercariae for their encystment, when snails live around the emergence of a source. The speed of the water current affected the number and distribution of metacercariae in the bed.

TITLE: Fasciola hepatica : la dispersion des cercaires émises par le mollusque Galba truncatula. ABSTRACT: Des investigations de terrain dans 14 cressonnières naturelles localisées dans la région française du Limousin, zone connue d'endémie de distomatose, ont été effectuées pendant trois années pour déterminer la distance que les cercaires de Fasciola hepatica peuvent atteindre dans l'eau avant leur enkystement sur la plante hôte. Chaque cressonnière était située sur le cours d'une rigole de drainage superficiel, tandis que les mollusques (Galba truncatula) vivaient plus en amont autour de l'émergence d'une source. Cinq espèces végétales ont été recueillies au début d'avril et examinées pour y trouver des métacercaires. La plupart des kystes ont été observés sur Nasturtium officinale (188 sur 48,7 kg de plantes égouttées), suivis par Helosciadium nodiflorum (125 sur 33,4 kg). Sur les autres espèces végétales, il y avait peu de larves. La plupart des cercaires étaient enkystées sur les plantes qui poussaient dans la partie la plus en amont de chaque cressonnière (habituellement sur les premiers 50 cm de longueur). Lorsque l'eau courante dans les cressonnières était rapide, la distribution des métacercaires était plus limitée et leur nombre était inférieur à celui observé dans les stations alimentées par un débit d'eau lent. Les cercaires ont été capables de nager ou ont été entrainées par le courant jusqu'à 5 m en moyenne dans une eau à écoulement lent avant leur enkystement ; cette distance n'était que de 4 m dans les eaux plus rapides. Les plantes qui poussent sur la section la plus en amont d'une cressonnière située dans une rigole de drainage sont les plus utilisées par les cercaires pour leur enkystement lorsque les mollusques vivent autour de l'émergence d'une source. La vitesse du courant d'eau affecte le nombre et la distribution des métacercaires dans la cressonnière.

Determination of zones at risk for fasciolosis in the department of Haute-Vienne, central France: a retrospective study on natural infections detected in 108,481 Galba truncatula for 37 years.

A retrospective study on the natural infection of Galba truncatula by Fasciola hepatica was carried out in the French department of Haute-Vienne to determine whether there are areas at risk for fasciolosis. Adult snails included in this analysis came from samples collected from pastures on 259 farms and from 121 wild watercress beds between 1970 and 2006. Fasciola hepatica infection rates were examined in relation to altitude and climatic data (mean annual rainfall, mean annual temperature) of each municipality. In a total of 108,481 snails collected in 151 municipalities, the overall prevalence of infection was 3.8% but varied according to the municipalities from which samples were taken (from 1% to 7.4%). The prevalence of F. hepatica infection in snails significantly decreased when the mean altitude of municipalities or their mean annual rainfall increased. However, this prevalence significantly increased with increasing mean annual temperatures. Studying the prevalence of infection in these snails makes it possible to delineate zones at risk for fasciolosis on the acid soils of Haute-Vienne. The risk of infection for livestock would be greater in areas of Haute-Vienne below 400 m above sea level and would gradually decrease when the altitude of the land increases.

TITLE: Détermination des zones à risque pour la fasciolose dans le département de la Haute-Vienne (France) : une étude rétrospective sur les infestations naturelles détectées chez 108.481 Galba truncatula pendant 37 années. ABSTRACT: Une étude rétrospective sur l'infestation naturelle de Galba truncatula par Fasciola hepatica a été effectuée dans le département de la Haute-Vienne pour déterminer s'il existe des zones à risque pour la fasciolose. Les limnées adultes impliquées dans cette analyse proviennent d'échantillons prélevés dans les pâturages de 259 fermes et dans 121 cressonnières sauvages entre 1970 et 2006. Les taux d'infection par Fasciola hepatica ont été mis en relation avec l'altitude et les données climatiques (précipitations annuelles moyennes, température annuelle moyenne) de chaque municipalité. Sur un total de 108.481 limnées récoltées dans 151 municipalités, la prévalence globale de l'infestation était de 3,8 %, mais variait selon les municipalités dans lesquelles les échantillons ont été prélevés (de 1 % à 7,4 %). La prévalence de l'infestation par F. hepatica diminue considérablement lorsque l'altitude moyenne des municipalités ou leurs précipitations annuelles moyennes augmente. D'autre part, cette prévalence augmente considérablement avec l'augmentation de la température annuelle moyenne des municipalités. L'étude de la prévalence de l'infestation chez ces limnées permet de délimiter des zones à risque pour la fasciolose sur les sols acides de la Haute-Vienne. Le risque d'infestation pour le bétail serait plus élevé dans les zones de la Haute-Vienne situées en dessous de 400 m d'altitude et diminuerait graduellement lorsque l'altitude de ces terrains augmente.

Galba truncatula and Fasciola hepatica: Genetic costructures and interactions with intermediate host dispersal.

Antagonistic interactions between hosts and parasites are key structuring forces in natural populations. Demographic factors like extinction, migration and the effective population size shape host-parasite metapopulational dynamics. Therefore, to understand the evolution of host-parasite systems it is necessary to study the distribution of the genetic variation of both entities simultaneously. In this paper, we investigate the population genetics co-structure of parasites and hosts within a metapopulation of the liver fluke, Fasciola hepatica, and two of its intermediate hosts, the main intermediate host in Europe, Galba truncatula, and a new intermediate host, Omphiscola glabra, in Central France. Our results reveal an absence of specificity of flukes as regard to the two alternative hosts though O. glabra shows higher prevalence of F. hepatica. Host and parasites displayed contrasting population genetics structure with very small, highly inbred (selfing) and strongly isolated G. truncatula populations and much bigger, panmictic and more dispersive F. hepatica. This could indicate a local adaptation of the parasite and a local maladaptation of the host. We also unveil a parasite-mediated biased population genetics structure suggesting that infected G. truncatula disperse more; have higher dispersal survival than uninfected snails or, more likely, that immigrant snails are infected more often than local snails (local parasites are less adapted to local hosts). Finally, an absence, or at least an ambiguous signature of isolation by distance was observed in both host and parasite population. A very weak migration rate for G. truncatula provides a reasonable explanation for this ambiguous result. Alternatively, smaller sample sizes combined with modest migration rates might explain the difficulties to unveil the signal in F. hepatica.

Current decline in the number and size of Galba truncatula and Omphiscola glabra populations, intermediate hosts of Fasciola hepatica, on the acidic soils of Central France.

Field investigations on the habitats colonized by Galba truncatula or Omphiscola glabra were carried out on 162 farms of the Limousin region, Central France, to determine whether there is currently a decline in the number and size of snail populations. Seven types of snail habitats were considered here. Compared to the numbers of snail populations recorded from 1976 to 1992, the values noted from 2013 to 2016 were significantly lower, with a decline rate of 34% for G. truncatula and 23% for O. glabra. Variations in this decline rate with the type of snail habitat were also noted. The greatest decreases in the numbers of snail populations were noted for spring heads located in meadows and for road ditches, while the lowest were noted for open drainage furrows present in meadows. The distribution of these habitats according to their area did not show any significant change over time. In contrast, overwintering snails were significantly less numerous in 2013-2016 in five types of habitats for G. truncatula and in three types only for O. glabra. Several causes underlie this population decline. Among them, the current development of mechanical cleaning in open drainage systems and road ditches, that of subsurface drainage in meadows, and regular gyro-crushing of vegetation around temporary spring heads were the most important.

Effect of Fascioloides magna (Digenea) on fecundity, shell height, and survival rate of Pseudosuccinea columella (Lymnaeidae).

Infection with Fascioloides magna (Digenea) causes serious damage to liver tissue in definitive hosts represented by ruminants, especially cervids. The distribution of F. magna includes the indigenous areas in North America, and the areas to which F. magna was introduced-Central Europe, Southeast Europe, and Italy. The North American intermediate host of F. magna, the freshwater snail Pseudosuccinea columella (Lymnaeidae), is an invasive species recorded in South America, the Caribbean, Africa, Australia, and west and Southeast Europe. In Europe, Galba truncatula is the snail serving for transmission, but P. columella has potential to become here a new intermediate host of F. magna. Little is known about interactions between F. magna and P. columella. In this study, the susceptibility of P. columella (Oregon, USA) to the infection by a single miracidium of the Czech strain of F. magna and the influence of F. magna on snail fecundity, shell height, and survival were evaluated. The data show that the Oregon strain of P. columella is a highly suitable host for the Czech strain of F. magna, with the infection rate of 74 %. In addition, a negative effect on survival rate of infected snails was recorded only in the late phase of infection. The infection was accompanied by a major reduction in egg mass production and by a decrease in the number of eggs per egg mass. The shell height of infected snails did not significantly differ from that in unexposed controls.

Larval trematode infections in Lymnaea glabra populations living in the Brenne Regional Natural Park, central France.

Lymnaea glabra is known to be a natural intermediate host of two flukes, Calicophoron daubneyi and Fasciola hepatica, in central France. But it can also sustain larval development of other digeneans. Adult snails were thus collected from 206 habitats in 2014 and 2015 to identify parasite species and determine the prevalence of each digenean infection in relation to the five types of snail habitats. Seven digenean species were noted in 321 infected snails (out of 17,647 L. glabra). Snails with F. hepatica or C. daubneyi were found in 14.5% and 12.6% of habitats, respectively. Percentages were lower for snails with Opisthoglyphe ranae (5.8%), Haplometra cylindracea (5.3%) and were less than 5% for those infected with Echinostoma revolutum, Notocotylus sp. or Plagiorchis sp. Prevalence noted for each parasite species varied with the type of habitat. The number of species in L. glabra was lower than that found in G. truncatula from the same region (7 instead of 10). The distribution and prevalence of each digenean species were thus dependent on the type and location of each snail habitat.

Fasciola hepatica: comparative metacercarial productions in experimentally-infected Galba truncatula and Pseudosuccinea columella.

As large numbers of metacercariae of Fasciola hepatica are necessary for research, experimental infections of Galba truncatula and Pseudosuccinea columella with this digenean were carried out to determine the better intermediate host for metacercarial production and, consequently, the most profitable snail for decreasing the cost price of these larvae. Pre-adult snails (4 mm in shell height) originating from two populations per lymnaeid species were individually exposed to two or five miracidia, raised at 23 °C and followed for cercarial shedding up to their death. Compared to values noted in G. truncatula, the survival of P. columella on day 30 post-exposure was significantly greater, while the prevalence of F. hepatica infection was significantly lower. In the four P. columella groups, metacercarial production was significantly greater than that noted in the four groups of G. truncatula (347-453 per cercariae-shedding snail versus 163-275, respectively). Apart from one population of G. truncatula, the use of five miracidia per snail at exposure significantly increased the prevalence of F. hepatica in P. columella and the other population of G. truncatula, whereas it did not have any clear effect on the mean number of metacercariae. The use of P. columella for experimental infections with F. hepatica resulted in significantly higher metacercarial production than that noted with G. truncatula, in spite of a lower prevalence for the former lymnaeid. This finding allows for a significant decrease in the cost price of these larvae for commercial production.

Pseudosuccinea columella: age resistance to Calicophoron daubneyi infection in two snail populations.

Individual infections of Egyptian and French Pseudosuccinea columella with five miracidia of Calicophoron daubneyi were carried out to determine whether this lymnaeid was capable of sustaining larval development of this parasite. On day 42 post-exposure (at 23 °C), infected snails were only noted in groups of individuals measuring 1 or 2 mm in height at miracidial exposure. Snail survival in the 2-mm groups was significantly higher than that noted in the 1-mm snails, whatever the geographic origin of snail population. In contrast, prevalence of C. daubneyi infection was significantly greater in the 1-mm groups (15-20% versus 3.4-4.0% in the 2-mm snails). Low values were noted for the mean shell growth of infected snails at their death (3.1-4.0 mm) and the mean number of cercariae (<9 in the 1-mm groups, <19 in the 2-mm snails). No significant differences between snail populations and snails groups were noted for these last two parameters. Most infected snails died after a single cercarial shedding wave. Both populations of P. columella showed an age resistance to C. daubneyi infection and only juveniles measuring 2 mm or less in shell height at exposure can ensure larval development of this digenean up to cercarial shedding.

Ruminal paramphistomosis in cattle from northeastern Algeria: prevalence, parasite burdens and species identification.

Slaughterhouse samples were analysed over a two-year period (September 2010-August 2012) in Jijel (northeastern Algeria) in order to determine seasonal variations in the prevalence and intensity of bovine paramphistomosis in a Mediterranean climate and identify paramphistome species using molecular biology. In spring and summer, significantly higher prevalences and lower parasite burdens were noted in bull calves, thus indicating an effect of season on these parameters. In contrast, the differences among seasonal prevalences or among seasonal parasite burdens were not significant in the case of old cows. Eleven adult worms from the slaughterhouses of Jijel and three neighbouring departments (Constantine, El Tarf and Setif) were analysed using molecular markers for species identification. Two different species, Calicophoron daubneyi and C. microbothrium, were found. The presence of these two paramphistomids raises the question of their respective frequency in the definitive host and local intermediate hosts.

Adaptation of Lymnaea fuscus and Radix balthica to Fasciola hepatica through the experimental infection of several successive snail generations.

BACKGROUND: High prevalence of Fasciola hepatica infection (>70%) was noted during several outbreaks before the 2000s in several French farms where Galba truncatula is lacking. Other lymnaeids such as Lymnaea fuscus, L. glabra and/or Radix balthica are living in meadows around these farms but only juvenile snails can sustain complete larval development of F. hepatica while older snails were resistant. The low prevalence of infection (<20%) and limited cercarial production (<50 cercariae per infected snail) noted with these juveniles could not explain the high values noted in these cattle herds. As paramphistomosis due to Calicophoron daubneyi was not still noted in these farms, the existence of another mode of infection was hypothesized. Experimental infection of several successive generations of L. glabra, originating from eggs laid by their parents already infected with this parasite resulted in a progressive increase in prevalence of snail infection and the number of shed cercariae. The aim of this paper was to determine if this mode of snail infection was specific to L. glabra, or it might occur in other lymnaeid species such as L. fuscus and R. balthica. METHODS: Five successive generations of L. fuscus and R. balthica were subjected to individual bimiracidial infections in the laboratory. Resulting rediae and cercariae in the first four generations were counted after snail dissection at day 50 p.e. (20°C), while the dynamics of cercarial shedding was followed in the F5 generation. RESULTS: In the first experiment, prevalence and intensity of F. hepatica infection in snails progressively increased from the F1 (R. balthica) or F2 (L. fuscus) generation. In the second experiment, the prevalence of F. hepatica infection and the number of shed cercariae were significantly lower in L. fuscus and R. balthica (without significant differences between both lymnaeids) than in G. truncatula. CONCLUSION: The F. hepatica infection of several successive snail generations, coming from parents infected with this parasite, resulted in a progressive increase in prevalence and intensity of snail infection. This may explain high prevalence of fasciolosis noted in several cattle-breeding farms when the common snail host of this digenean, G. truncatula, is lacking.

Radix natalensis: the effect of Fasciola hepatica infection on the reproductive activity of the snail.

Experimental infections of Egyptian Radix natalensis (shell height at miracidial exposure: 4 mm) with a French isolate of Fasciola hepatica were carried out under laboratory conditions at 22 °C to specify the characteristics and follow the dynamics of their egg-laying. Controls constituted unexposed R. natalensis of the same size. No significant difference between controls and the uninfected snails of the exposed group was noted, whatever the parameter considered. In controls and exposed snails, the dates of the first egg masses were close to each other (56.4-65.3 days). In contrast, the life span of snails and the length of the egg-laying period were significantly shorter and egg production was significantly lower in infected R. natalensis than in controls and uninfected snails. In infected R. natalensis, but without cercarial shedding (NCS snails), egg production was irregular throughout the egg-laying period. In cercarial-shedding (CS) snails, the first egg masses were laid before the first cercarial emergence (at a mean of 56 days and 67 days, respectively). Thereafter, egg mass production of CS snails was irregular up to day 72 of the experiment, stopped during the following two weeks and started again after day 88 for a single snail. In conclusion, the F. hepatica infection of R. natalensis reduced the reproductive activity in both NCS and CS snails. The pattern noted for egg production in infected R. natalensis seems to be species-specific because of the high shell size of this lymnaeid and its role as an atypical intermediate host in the life cycle of the parasite.

Lymnaea cubensis, an experimental intermediate host for Fascioloides magna.

Single-miracidium infections of Lymnaea cubensis (Pfeiffer) from Guadeloupe with the giant liver fluke Fascioloides magna (Bassi, 1875) (Digenea) were carried out during five successive snail generations to determine if this lymnaeid might sustain complete larval development of the parasite. Controls were constituted by a French population of Galba truncatula (Miller) (a single generation) infected according to the same protocol. It was recorded that prevalence and intensity of F. magna infection in L. cubensis progressively increased from F1 to F5 generations. Cercarial shedding of F. magna was noted only within F5 generation of L. cubensis. However, most measured parameters of infection in this species were significantly lower than those noted for G. truncatula and most L. cubensis died after a single shedding wave. Despite this, L. cubensis can be added to the list of potential intermediate hosts of F. magna.

Fasciola hepatica: effect of the natural light level on cercarial emergence from temperature-challenged Galba truncatula.

As abrupt changes in water temperature (thermal shock) triggered a significantly greater cercarial emergence of Fasciola hepatica from experimentally infected Galba truncatula, laboratory investigations were carried out to study the influence of light on cercarial emergence in snails subjected to a thermal shock every week (a mean of 12 °C for 3 h) during the patent period. Thermal shock for these temperature-challenged (TC) snails was carried out outdoors under artificial or natural light, or indoors under constant artificial light. Compared with the infected control snails always reared indoors at 20 °C, the number of cercariae in TC snails subjected to a thermal shock and natural light outdoors was significantly greater. The repetition of this experiment by subjecting TC snails to the same thermal shock indoors under an artificial light level ranging from 600 to 3000 lux did not show any significant difference among the numbers of cercariae in the different subgroups. A detailed analysis of the results noted in the TC snails subjected to natural light during the thermal shock demonstrated that the number of cercariae-releasing snails was significantly higher between 601 and 1200 lux and for the highest nebulosity values (7-8 octas, which corresponds to a sufficiently or completely overcast sky). Contrary to the intensity of artificial light, which did not influence cercarial emergence, the natural light level had a significant effect on this process when F. hepatica-infected snails were subjected to a regular thermal shock during the patent period.

Fasciola hepatica: variations in redial development and cercarial production in relation to the geographic origin of the parasite.

Two hundred Galba truncatula, originating from a French population, were subjected to single-miracidium infections with an allopatric isolate (origin, Argentina) of Fasciola hepatica. The control group was constituted by 100 snails coming from the same population and exposed to sympatric miracidia of F. hepatica according to the same protocol. Snail samples were collected bimonthly from both groups between day 14 and day 112 p.e. (at 20 °C) and snail dissections were performed to count free rediae, intraredial morulae and free cercariae. Third and fourth generation rediae were significantly more numerous in the allopatric group, while the number of first generation rediae was significantly lower. In the sympatric group, the decrease in the number of intraredial morulae in the first, second and third redial generations was significantly faster. Free cercariae within the snail body were significantly more numerous in the sympatric than in the allopatric groups, whatever the date of snail dissection. The changes in redial development and cercarial production noted in the Argentinean group might be due to the evolution of South American flukes in a divergent way after the introduction of foreign infected ruminants in this continent from the 15th century.

Lymnaea palustris and Lymnaea fuscus are potential but uncommon intermediate hosts of Fasciola hepatica in Sweden.

BACKGROUND: Lymnaea palustris and L. fuscus are members of the European stagnicolines (Gastropoda: Lymnaeidae). The role of stagnicolines in transmission of Fasciola hepatica has been often proposed. To assess the possible relationship between these two stagnicolines and F. hepatica in Sweden, field monitoring in parallel with experimental infections of L. palustris and L. fuscus were conducted. METHODS: Stagnicoline snails were collected and identified on pastures grazed by either sheep or cattle on four farms suffering from fasciolosis in Sweden during 2011-2012. Field-collected L. palustris and L. fuscus were examined for F. hepatica DNA by PCR. In the laboratory, different age groups of L. palustris, L. fuscus and G. truncatula were each exposed to two F. hepatica miracidia and main infection characteristics were obtained. RESULTS: One field-collected L. palustris (out of n = 668) contained F. hepatica as determined by PCR. On the other hand, stagnicolines artificially exposed to F. hepatica miracidia resulted in successful infection with fully differentiated cercariae, but only in juvenile snails (size, 1-2 mm at exposure) and with a prevalence of 51% and 13% in L. palustris and L. fuscus, respectively. In contrast, 90% of juvenile (size, 1-2 mm) and 92% of preadult G. truncatula (size, ≥ 2-4 mm), respectively, were successfully infected. Delayed, reduced and/or no spontaneous cercarial shedding was observed in the two stagnicolines when compared to G. truncatula. However, at snail dissection most cercariae from L. fuscus and L. palustris were able to encyst similarly to those from G. truncatula. CONCLUSION: Both L. fuscus and L. palustris can sustain larval development of F. hepatica but with an apparent level of age resistance. The finding of a single F. hepatica positive specimen of L. palustris, together with infection characteristics from the experimental infection, suggest that L. palustris is a more suitable snail vector of F. hepatica than L. fuscus. The reduced growth observed in both stagnicolines was contrary to the 'parasitic gigantism' theory. Overall, it seems that the epidemiological role of L. palustris in transmission of F. hepatica in Sweden is likely to be much lower than for G. truncatula.

Consequence of temperature changes on cercarial shedding from Galba truncatula infected with Fasciola hepatica or Paramphistomum daubneyi.

Experimental infections of Galba truncatula (two populations) with Fasciola hepatica or Paramphistomum daubneyi were carried out to study the effect of water temperature changes (3 h at a mean of 12 °C every week) on cercarial shedding during the patent period. The results were compared with those of control snails infected according to the same protocol and always maintained at 20 °C. Compared to controls, a significant increase in the number of cercariae-shedding snails, a significantly longer patent period and significantly greater cercarial production were noted in temperature-challenged snails, regardless of the type of digenean infection. In contrast, the number of incompletely formed metacercariae was significantly higher in temperature-challenged snails than in controls. Incompletely formed metacercariae of F. hepatica consisted of cysts whose colour remained whitish after shedding (25.4% for temperature-challenged snails) or whose dome was flattened after encystment (74.6%). Those of P. daubneyi were totally dark brown or blackish after formation. These incomplete metacercariae might originate from young differentiating cercariae within the snail body (F. hepatica) or from cercariae which died just after encystment (P. daubneyi). The use of regular temperature changes for snails infected with F. hepatica or P. daubneyi must be monitored carefully during collection of metacercariae to select completely formed cysts for infecting definitive hosts.

Lymnaea fuscus (Pfeiffer, 1821) as a potential intermediate host of Fascioloides magna in Europe.

Experimental infections of two different populations of Lymnaea fuscus in France and Sweden, with a Czech isolate of Fascioloides magna were carried out to determine if this lymnaeid species enables parasite larval development. Species identification of both snail populations was performed using the morphology of the copulatory organ, and also confirmed by sequencing of the internal transcribed spacer 2 (ITS2) region of the snail genomic rDNA. Only juvenile snails measuring less than 3mm (1-3 weeks of age) were successfully infected (the viable cercariae were recorded) and infection prevalence decreased with age, as documented by increased shell height. In both French and Swedish L. fuscus populations, prevalence ranged between 1.1% and 58.8%. The mean number of metacercariae obtained from cercariae-shedding snails was 13.7 (±11.4), while the total cercarial production noted in snails dissected at day 85 post-exposure was 147.5 (±56.6). Compared to uninfected control snails, we observed reduced growth of infected snails. Despite age-related resistance of snail to the parasite, and limited cercarial production in these experimentally infected snails, F. magna was still able to complete larval development in L. fuscus.

The detection of snail host habitats in liver fluke infected farms by use of plant indicators.

Field investigations in 361 liver fluke infected cattle- or sheep-breeding farms on acid soil were carried out during thirty years in March and April to record indicator plants in relation to the category of site colonized by the intermediate host of liver fluke, the snail Galba truncatula. Seven types of snail zones and six species of indicator plants were recorded in the 7709 positive sites studied. The most frequent habitats were located at the peripheral extremities of open drainage furrows. Juncus acutiflorus, Juncus effusus, Glyceria fluitans, and Agrostis stolonifera were the indicator plants. Plant indicators were highly efficient (38-80% of variance explained) when used with the size of site area within a hydrographical zone. The identification of positive sites for G. truncatula is a key for controlling the intermediate host on pastures through biological control with predatory molluscs. The positive sites may be detected using a two-step method including first categorization of hydrographical zones and then, within a zone, use of one or several indicator plants (frequently J. acutiflorus).

The detection and quantification of a digenean infection in the snail host with special emphasis on Fasciola sp.

In this review, ten methods used to study digenean infections in their intermediate hosts were compared to determine which one should be used either in the field or in the lab to establish the prevalence and intensity of infections in snails. Snail crushing and snail dissection allow quick establishing of prevalence in natural or experimental infections, whereas histology is considered as the most accurate approach to assess the intensity of infection. The follow-up of cercarial shedding only gave an idea on cercarial production. Among recently developed techniques, polymerase chain reaction (PCR) brings the most accurate information and shows high sensitivity and specificity levels when compared to blotting techniques. The easiness and relatively low cost of the basic PCR protocol make it interesting to investigate the epidemiology of the liver fluke in a lab with limited financial resources. Nevertheless, if this technique allows a relatively good estimation of the prevalence, information concerning the intensity of infection is best obtained through real time PCR. However, at the time being this technique is too expensive to be used routinely in the field. The choice between classical or new techniques is usually based on a compromise, as each technique has its advantages and drawbacks.