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Four freshwater pulmonate species (Lymnaea ovata, L. stagnalis, Physa acuta, Planorbis leucostoma)were living in several watercress beds known for their relationships with human cases of fasciolosis, whereas L. truncatula was never found. The aims of these studies were to determine the prevalence of natural infections with Fasciola hepatica in snails and to verify if these species might ensure the full larval development of this trematode (with cercarial shedding] when they were experimentally subjected to F. hepatica only, or to co-infections with an other trematode species. Investigations were so carried out in six snail populations living in watercress beds (including three for P. acuta) and in four others originating from three brooks or a pond (as controls). Snails naturally infected with F. hepatica were found in two watercress beds inhabited by L. ovata (prevalence of infection: 1.4 %) and P. leucostoma (0.1 %), respectively. The L. ovata from the watercress bed could be infected at a higher size than those from the control population and the prevalence of this infection was greater in the bed population. Similar findings were noted for L. stagnalis. Despite single or dual infections, the results obtained with the four populations of P. acuta were unsuccessful. In contrast, the co-infections of young P. leucostoma with Paramphistomum daubneyi and F. hepatica resulted in the shedding of some F. hepatica cercariae. According to the authors, the occurrence of fasciolosis in these watercress beds would be the consequence of frequent natural encounters between parasite and snails (L. ovata, L. stagnalis), or of co-infections with P. daubneyi and F. hepatica (P. leucostoma). In watercress beds only colonized by P. acuta, a lymnaeid species would have ensured the larval development of F. hepatica but it would have been eliminated by P. acuta, as this last species was known to be invasive and could colonize open drainage ditches on siliceous soil.

Lymnaeids in nature are subjected to temperature fluctuations that may have an influence on the shedding of cercariae. Thus, experimental infections of Lymnaea truncatula by Paramphistomum daubneyi were performed to determine whether a sudden fall in temperature--daily dipping of infected snails into spring water at 14-15 degrees C, or at 6-8 degrees C--followed by its increase at 20 degrees C in the subsequent hour, had an influence on the characteristics of snail infection and cercarial production. The immersion of infected snails in cold water during a short period delayed the first cercarial shedding, at day 66 on average, in the 6-8 degrees C group vs at day 57 in the 14-15 degrees C group, or at day 49 in the 20 degrees C group. The percentage of cercaria-shedding snails was greater in the 6-8 degrees C group than in the 14-15 degrees C and the 20 degrees C ones: 41.8% vs 17.3% and 7.1%, respectively. The total number of cercariae given by each infected snail was also higher in the 6-8 degrees C group. A fall in the temperature of water during daily change, followed by its increase at 20 degrees C within the subsequent hour, stimulated the cercarial shedding of P. daubneyi.

Three experiments on the infection of Lymnaea fuscus with Fasciola hepatica were carried out to determine if successful infections and maturation of the parasite were dependent on the size of snails at miracidial exposure. The first experiment was performed using 1-4-mm-high snails from 2 populations of L. fuscus and 1 population of Lymnaea palustris. In these snails each subjected to a single bimiracidial exposure, the prevalence of F. hepatica infection at day 35 postexposure ranged from 20.3% to 46.2% in snails measuring 1 mm in height at exposure; it was lower in the 2-mm snails and was 0 in higher size classes. The second experiment was performed by subjecting 1- and 4-mm L. fuscus to 1, 2, and 3 bimiracidial exposures. The prevalence of F. hepatica infection at day 35 postexposure was maximum in the 1-mm snails exposed once to miracidia and decreased with increasing number of exposures. The results were negative in 4-mm snails. Cercarial shedding of F. hepatica was studied in the third experiment using 1- and 2-mm L. fuscus each subjected to a single bimiracidial exposure. The total number of cercariae released from these snails was less than 50. From these results, it can be concluded that L. fuscus showed a partial resistance to F. hepatica infection due to snail age.

Natural infection of Lymnaea glabra and Lymnaea truncatula by Fasciola hepatica and/or Paramphistomum daubneyi was studied at two periods of risk (June-July and September-October) in 11 French farms known for their high prevalences of F. hepatica infection in ruminants. A total of 1,778 L. truncatula and 2,396 L. glabra measuring 6 mm or more in height were collected to determine the prevalence of natural infection with F. hepatica and P. daubneyi. The role of four factors, i.e. season, snail co-infection (F. hepatica-P. daubneyi), ruminant host and the existence of single or mixed intermediate host communities, was investigated. There were no differences in prevalences between the two risk periods. Co-infections in L. glabra were more frequent than expected. The prevalences of infection with F. hepatica or P. daubneyi in L. glabra were higher in farms rearing sheep than in farms rearing cattle. A similar finding was also noted for L. truncatula infected with P. daubneyi only. L. glabra was a much better intermediate host for F. hepatica and P. daubneyi when it was the only available snail, possibly indicating an adaptation of parasites to their less usual host in local conditions. The two trematodes preferably developed in L. truncatula rather than in L. glabra when both host species lived in the same places.

Cases of fasciolosis in ruminants have been recorded in several French farms in the absence of Lymnaea truncatula, which is considered the only snail intermediate host in western Europe. These farms harbored other species of freshwater snails in large numbers (Lymnaea glabra, Physa acuta, or Planorbis leucostoma) and, in many cases, had cattle or sheep infected by another trematode (Paramphistomum daubneyi). These other freshwater snails may serve as intermediate hosts for F. hepatica due to a coexisting infection with P. daubneyi. We have demonstrated that L. glabra, either infected with F. hepatica alone or coinfected by P. daubneyi, was capable of developing a F. hepatica infection. A broader range of L. glabra size classes (up to 10 mm in height) were susceptible to infection if simultaneously infected with P. daubneyi. Planorbis leucostoma can only serve as an intermediate host for F. hepatica, if infected with P. daubneyi. Lastly, P. acuta smaller than 4 mm cannot serve as an intermediate host. These results may explain, in part, the maintenance of low-level F. hepatica infections in the absence of the normal intermediate host, L. truncatula.

Preadult Lymnaea glabra measuring 4-6 mm in height were each exposed to 1 Paramphistomum daubneyi miracidium before being exposed to 1 miracidium of Fasciola hepatica. Total prevalence of infection in the snail groups from 3 different populations ranged from 33% to 39%. In each group, snails harboring larval forms of P. daubneyi, F. hepatica, or both, were noted. If the results from the 3 snail populations are pooled, the total prevalence of snail infection was 13.6% in snails harboring only F. hepatica larvae, 13% in those harboring only P. daubneyi larvae, and 10% in those with both trematodes. Cercarial shedding was obtained from snails harboring F. hepatica larval forms; in the case of snails infected with both trematodes, the mean number of metacercariae ranged from 13 to 21 for F. hepatica, and from 8 to 14 for P. daubneyi. No infected snails were found in snail groups exposed only to 1 trematode miracidia.