First report of Melanoides tuberculata (Mollusca: Thiaridae) harboring a xiphidiocercaria in Brazil: A new parasite introduced in the Americas?

Xiphidiocercariae were found in the invasive snail Melanoides tuberculata collected during a malacological survey in Ceará-Mirim, State of Rio Grande do Norte, Northeastern Brazil in November 2018 and submitted to morphological and molecular analyses. The morphology revealed similarities between the larvae here reported for the first time in M. tuberculata from Brazil and other xiphidiocercariae described in thiarid snails from Asia and Africa. Phylogenetic analyses based on 28S and ITS-2 sequences revealed that the larvae correspond to an unidentified species of the family Lecithodendriidae. Aspects related to the morphology and taxonomy of xiphidiocercariae found in M. tuberculata are briefly discussed. It is possible that the parasite here reported is a newly introduced species transmitted by M. tuberculata in the American continent.     

Morphological and Molecular Characterization of Lymnaeid Snails and Their Potential Role in Transmission of Fasciola spp. in Vietnam

Freshwater snails of the family Lymnaeidae play an important role in the transmission of fascioliasis worldwide. In Vietnam, 2 common lymnaeid species, Lymnaea swinhoei and Lymnaea viridis, can be recognized on the basis of morphology, and a third species, Lymnaea sp., is known to exist. Recent studies have raised controversy about their role in transmission of Fasciola spp. because of confusion in identification of the snail hosts. The aim of this study is, therefore, to clarify the identities of lymnaeid snails in Vietnam by a combination of morphological and molecular approaches. The molecular analyses using the second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA clearly showed that lymnaeids in Vietnam include 3 species, Austropeplea viridis (morphologically identified as L. viridis), Radix auricularia (morphologically identified as L. swinhoei) and Radix rubiginosa (morphologically identified as Lymnaea sp.). R. rubiginosa is a new record for Vietnam. Among them, only A. viridis was found to be infected with Fasciola spp. These results provide a new insight into lymnaeid snails in Vietnam. Identification of lymnaeid snails in Vietnam and their role in the liver fluke transmission should be further investigated.     

Release of Lungworm Larvae from Snails in the Environment: Potential for Alternative Transmission Pathways

Background

Gastropod-borne parasites may cause debilitating clinical conditions in animals and humans following the consumption of infected intermediate or paratenic hosts. However, the ingestion of fresh vegetables contaminated by snail mucus and/or water has also been proposed as a source of the infection for some zoonotic metastrongyloids (e.g., Angiostrongylus cantonensis). In the meantime, the feline lungworms Aelurostrongylus abstrusus and Troglostrongylus brevior are increasingly spreading among cat populations, along with their gastropod intermediate hosts. The aim of this study was to assess the potential of alternative transmission pathways for A. abstrusus and T. brevior L3 via the mucus of infected Helix aspersa snails and the water where gastropods died. In addition, the histological examination of snail specimens provided information on the larval localization and inflammatory reactions in the intermediate host.

Methodology/Principal Findings

Twenty-four specimens of H. aspersa received ~500 L1 of A. abstrusus and T. brevior, and were assigned to six study groups. Snails were subjected to different mechanical and chemical stimuli throughout 20 days in order to elicit the production of mucus. At the end of the study, gastropods were submerged in tap water and the sediment was observed for lungworm larvae for three consecutive days. Finally, snails were artificially digested and recovered larvae were counted and morphologically and molecularly identified. The anatomical localization of A. abstrusus and T. brevior larvae within snail tissues was investigated by histology. L3 were detected in the snail mucus (i.e., 37 A. abstrusus and 19 T. brevior) and in the sediment of submerged specimens (172 A. abstrusus and 39 T. brevior). Following the artificial digestion of H. aspersa snails, a mean number of 127.8 A. abstrusus and 60.3 T. brevior larvae were recovered. The number of snail sections positive for A. abstrusus was higher than those for T. brevior.

Conclusions

Results of this study indicate that A. abstrusus and T. brevior infective L3 are shed in the mucus of H. aspersa or in water where infected gastropods had died submerged. Both elimination pathways may represent alternative route(s) of environmental contamination and source of the infection for these nematodes under field conditions and may significantly affect the epidemiology of feline lungworms. Considering that snails may act as intermediate hosts for other metastrongyloid species, the environmental contamination by mucus-released larvae is discussed in a broader context.     

Paramphistomum daubneyi: characteristics of infection in three lymnaeid species

Experimental infections of two South American lymnaeids (Lymnaea neotropica and L. viatrix var. ventricosa) with Paramphistomum daubneyi were carried out to determine if these snail species could sustain larval development of this digenean and, if so, to specify their potential for cercarial production. A French population of Galba truncatula infected and raised according to the same protocol served as controls. In both experiments, prevalence of P. daubneyi infections in snails did not significantly differ from each other. In snail groups evaluated for cercarial shedding (first experiment), a significantly lower number of shed cercariae was noted for L. neotropica, while those from G. truncatula and L. v. ventricosa did not differ significantly from each other. Dissection of infected snails at day 65 post-exposure at 20 °C (second experiment) found significantly lower burdens of P. daubneyi rediae and cercariae in the bodies of L. neotropica than in those of G. truncatula and L. v. ventricosa. Compared to total cercarial production observed in dissected snails, the percentage of cercariae which exited from snails was 75.6% for G. truncatula, 21.6% for L. neotropica, and 91.4% for L. v. ventricosa. This last species seems to be a good candidate for metacercarial production of P. daubneyi.     

Phylogenetics,biogeography, and evolutionary trends of the Phalaenopsis sumatrana complex inferred from nuclear and chloroplast DNA

Phylogenetic trees inferred from the internal transcribed spacers 1 and 2 (ITS1 + ITS2) region from nuclear ribosomal DNA (nrDNA) and the intergenic spacer of atpB-rbcL from chloroplast DNA (cpDNA) were used to clarify the phylogenetics and evolutionary trends of the Phalaenopsis sumatrana complex. The P. sumatrana complex includes the two species P. sumatrana and Phalaenopsis corningiana as well as a problem species, Phalaenopsis zebrina, according to the concepts of Sweet (1980) and Christenson (2001). Based on the phylogenetic tree inferred from the ITS sequence, the accessions of P. sumatrana cannot be separated from those of P. corningiana. In contrast, the accessions of P. zebrina can be separated from those of both P. sumatrana and P. corningiana. However, analyses of the sequences of the atpB-rbcL spacer apparently cannot discriminate among these three species of the P. sumatrana complex. An inspection of the morphological characters of the plants of the P. sumatrana complex and the floral fragrances of P. zebrina can be used to separate it from both P. sumatrana and P. corningiana. Based on the molecular data and floral fragrances, P. zebrina perhaps should not be treated as a synonym of P. sumatrana. On the evolutionary trend of the P. sumatrana complex, P. zebrina was suggested to be the relative origin group of the P. sumatrana complex based on the phylogenetic tree and biogeography.     

Rhabditis maupasi: occurrence in food snails and cultivation

Rhabditis maupasi, a nematode, inhabits the mantle cavity of terrestrial snails, Helix aspersa, that are imported from Morocco and sold in the food markets of New York City. Most of the nematodes found in living snails are stage-3 larvae which mature and reproduce after the snail has died and decayed. The nematodes were isolated from snails and cultivated with a microflora and then in species isolation, i.e., axenically. Sterile slices of rabbit kidney supported by agar slants served as the initial culture system. Subsequently, a liquid medium was used that contained raw rabbit liver extract. When, instead, raw snail extract was substituted in the liquid medium, nematode larvae did not mature unless the snail extract was digested secondarily or inactivated by heating. Nematodes grow abundantly in dead snails and in treated snail extract but not in living snails or in raw snail extract. The reason is probably not the nematodes' lack of suitable digestive enzymes for snail tissue nor the absence of adequate nutrients in living, microbially poor snails, but rather the presence of an inhibitor in snails.     

Effect of starvation on parasite-induced mortality in a freshwater snail (Potamopyrgus antipodarum)

The level of host exploitation is expected, under theory, to be selected to maximise (subject to constraints) the lifetime reproductive success of the parasite. Here we studied the effect of two castrating trematode species on their intermediate snail host, Potamopyrgus antipodarum. One of the trematode species, Microphallus sp., encysts in the snail host and the encysted larvae “hatch” following ingestion of infected snails by birds. The other species, Notocotylus gippyensis, by contrast, releases swimming larvae; ingestion of the snail host is not required for, and does not aid, transmission to the final host. We isolated field-collected snails for 3 months in the laboratory, and followed the survival of infected and uninfected snails under two conditions: not fed and fed ad libitum. Mortality of the infected hosts was higher than mortality of the uninfected ones, but the response to starvation treatment was parasite species specific. N. gippyensis induced significantly higher mortality in starved snails than did Microphallus. Based on these results, we suggest that host exploitation by different species of trematodes may depend on the type of transmission. Encysting in the snail host may select for a reduced rate of host exploitation so as to increase the probability of transmission to the final host. Received: 29 July 1998 / Accepted: 1 February 1999     

The effect of some biological and physical factors on infection of Biomphalaria glabrata with Angiostrongylus cantonensis.

Several biological and physical factors which may influence infection of Biomphalaria glabrata snails with the first stage larvae of Angiostrongylus cantonensis were studied. These factors were: the size of snails, the number of first stage larvae to which snails were exposed, the age of larvae, individual exposure compared with mass exposure of snails, the length of exposure period and the effect of temperature and light. The results showed that young snails, 2 mm in diameter, became infected with significantly smaller numbers of third stage larvae than larger snails (4, 8, 12 and 16 mm). No significant differences in the intensity of infection were evident between the larger size groups. The number of third stage larvae recovered from snails was directly related to the number of first stage larvae to which the snails were exposed. The mean percentage recovery per snail was more or less constant regardless of the infective dose. First stage larvae showed a slight reduction in their survival rate within 7 and 4 days, at 24 degrees C and 26 degrees C respectively, after which a sharp decrease in their survival rate occurred. However, the infectivity of larvae was progressively reduced from the second day at these two temperatures. The mean intensity of infection in snails was independent of whether the infection was by mass or individual exposure. The average number of first stage larvae entering a snail was greatest during the first half hour of exposure, this decreased considerably during the second half hour, and thereafter the number of larvae which entered a snail was low. It was concluded that 26 degrees C was the optimal temperature for infection and that the presence or absence of light had no effect on the infection.     

Population dynamics of Melanoides tuberculata (Thiaridae) snails in a desert spring,United Arab Emirates and infection with larval trematodes

A dense population of Melanoides tuberculata was found at a brackish (5.93–6.98%) desert spring in the United Arab Emirates. A total of 232–300 snails were measured and examined for larval trematodes during the period January 1990 to January 1991. The relative abundance of various size classes of the snail has shown that the mass release of young snails occurred twice a year, in April–May and September–October. Seven different forms of cercariae were found: A xiphidiocercaria, a brevifurcate cercaria, two pleurolophocercous cercariae, two gymnocephalous cercariae, and the cercaria of Philophthalmus gralli. The overall infection rate of this snail was 73.6% and ranged from 51.3 during February–March 1990 to 91.7% during January 1991. The xiphidiocercaria and the P. gralli cercaria were the most abundant and were found in 21.1% and 29.7%, respectively, of the examined snails. A relatively high rate (9.5%) of double infection was found. Cases of triple infection were also encountered.     

Eye trematode infection in small passerines in Peru caused by Philophthalmus lucipetus,an agent with a zoonotic potential spread by an invasive freshwater snail

Until now, four species of eye trematodes have been found in South America. Of them, Philophthalmus lucipetus (synonymized with Philophthalmus gralli) displays a broad host spectrum, with at least 30 bird species (prevalently large water birds), five mammal species and humans serving as definitive hosts, and with snails Fagotia (Microcolpia) acicularis, Amphimelania holandri, Melanopsis praemorsa and Melanoides tuberculata serving as intermediate hosts. When examining a total of 50 birds of ten species in the wetland of Pantanos de Villa, Lima, Peru in July 2011, eye trematodes were identified visually in the edematous conjunctival sac of 11 (48%) out of 23 resident many-colored rush tyrants Tachuris rubrigastra. Based on morphometric characteristics, the trematodes were identified as P. lucipetus. ITS2 and CO1 gene of the examined specimens combined showed a 99% similarity to an Iranian isolate of Philophthalmus sp. from the intermediate host Melanoides tuberculata, an invasive freshwater snail, suggesting that these two isolates represent the same species with a wide geographical range. Moreover, the prevalence of infection with the philophthalmid cercariae was 31% in 744 Melanoides tuberculata examined in Pantanos de Villa in 2010. It is evident that P. lucipetus occurs throughout the world as well as locally, including Eurasia and South America. Here we report this trematode for the first time in Peru, and we were the first to sequence any of the South American eye trematodes. Low host specificity of P. lucipetus and the invasive character of Melanoides tuberculata as a competent intermediate host suggest that eye trematodosis caused by P. lucipetus may emerge frequently in various parts of the world, especially in the tropics. Increase of the zoonotic potential of the P. lucipetus associated with this invasive snail spreading across the world is predictable and should be of interest for further research.     

Studies on the prolonged storage of Metarhizium anisopliae conidia: effect of temperature and relative humidity on conidial viability and virulence against mosquitoes

The shell lengths, dried shell weights, soft tissue wet weights, and soft tissue dry weights were ascertained for noninfected Ilyanassa obsoleta and for the same snail species naturally infected with the following digeneans: Himasthla quissetensis, Zoogonus lasius, Cercaria dipterocerca, Lepocreadium setiferoides, Microphalloides nassicola, Stephanostomum tenue, and Microbilharzia variglandis. Analyses of the data obtained indicate that infection with all of the trematodes listed above causes neither enhanced growth of soft tissues nor accelerated lengthening of the shell. However, snails infected with sporocysts of Z. lasius have significantly heavier shells than do noninfected snails. These data suggest that infection with Z. lasius may have caused enhanced growth in the form of increased calcium deposition in the shell of I. obsoleta, possibly as a result of parasitic castration. In addition, the mean dry weight of the soft tissues of snails infected with M. variglandis, when normalized for shell length, is significantly lower than that of noninfected snails, possibly as a result of parasite-induced pathology. Finally, comparisons among infected snails indicate a trend toward increased soft tissue dry weight in snails infected with L. setiferoides, although the mean dry weight of these snails does not differ significantly from the mean dry weight of noninfected snails.     

Total protein and carbohydrate content and protease and disaccharidase activities in the hemolymph of Lymnaea stagnalis naturally infected with digenean larvae

Lymnaea stagnalis is an intermediate host of many Digenea. The infestation affects host metabolism. The aim of the work was to investigate hemolymph biochemical indicators of L. stagnalis infected with four species of trematodes: Diplostomum pseudospathaceum, Paryphostomum radiatum, Plagiorchis elegans or Opisthioglyphe ranae. The protein profiles and proteinase activity in the hemolymph of sexually mature individuals of Lymnaea stagnalis maintained at 19°C were tested. As the carbohydrates are main substrates for energetic metabolism of the great pond snail their content and disaccharidase activity were also studied. Hemolymph samples were collected during weeks 3 and 4 of rearing. No significant differences in the total protein content between uninfected individuals and snails infected with the first three trematode species were detected. In the snails infected with O. ranae the quantity of total proteins was near twice higher than in those uninfected. A higher share of 70 kDa proteins in infected than in uninfected snails as well as reduction of the low molecular weight fractions of proteins for snails infected with D. pseudospathaceum and P. radiatum were detected. During week 3, carbohydrate content in the infected snails did not differ from that in the controls while during week 4 it was significantly lower in snails infected with P. elegans or O. ranae. The content of the major soluble carbohydrate in the hemolymph - saccharose — changed in a similar way. No activity of trypsin or pepsin in the hemolymph sample was detected while the activity of chymotrypsin was lower in infected snails vs. controls. On the other hand, saccharase and maltase activities were higher in infected than in uninfected snails. The biochemical hemolymph indicators in naturally infected host-snails show some differences depending on the parasite species but they are not sufficiently species-specific to offer the basis for establishing the model unique for a particular parasitosis. Our results from the field did not always coincide with those from the laboratory.     

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 3 mm (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.     

Tissue responses exhibited by Biomphalaria alexandrina snails from different Egyptian localities following Schistosoma mansoni exposure

Snails’ susceptibilities to infection with Schistosoma mansoni were determined through observation of infection rates, total cercarial production and tissue responses of the first generation (F1) of Biomphalaria alexandrina snails, originally collected from different Egyptian governorates (Giza, Fayoum, Kafr El-Sheikh, Ismailia and Damietta) and responses were compared between groups. The emergence of cercariae for a 3-month period and the calculation of survival and infection rates, in control (Schistosome Biological Supply Center; SBSC) and infected snails were evaluated. SBSC and Giza snails showed greater susceptibilities to infection and lower mortality rates. In addition, at 6 and 72 h post-exposure to miracidia all the snail groups showed no difference in the anatomical locations of sporocysts. The larvae were found in the head-foot, the mantle collar and the tentacles of the snails. Sporocysts showed normal development with low tissue reactions in SBSC and Giza snail groups infected with S. mansoni miracidia (SBSC). However, in Fayoum, Kafr El-Sheikh, Ismailia and Damietta snail groups, variable tissue responses were observed in which numerous hemocytes made direct contact with S. mansoni larvae forming capsules. The results suggested that, different responses of B. alexandrina snail’s hemocytes towards S. mansoni are related to the degree of susceptibility of these snails. So this is important in planning the strategy of schistosomiasis control.     

Schistosoma incognitum from Cikurai,West Java,Indonesia

Schistosoma incognitum is reported for the first time from West Java, Indonesia where 84% of ricefield rats, Rattus argentiventer, were infected. Radix auricularia rubiginosa snails were intermediate hosts. The adult stage of the Javanese strain of S. incognitum and lesions in naturally-infected rodents are described. Rattus exulans, Rattus norvegicus and Mus musculus were experimentally infected. Epizootiology is similar to that described in other countries.     

Focal Philophthalmus gralli infection possibly persists in Melanoides tuberculata over two years following the definitive hosts' removal

Philophthalmosis is a zoonotic disease associated largely with the spread of the invasive freshwater snail Melanoides tuberculata, serving as an intermediate host. Here we examined Philophthalmus gralli focal fenced infection site reported recently as being associated with Tinamus major and M. tuberculata in Alajuela, Costa Rica. Removal of the definitive hosts allowed us to address also the long-term survival strategy of the parasite. Initially, the snail intermediate hosts displayed high prevalence of P. gralli infection across all its age cohorts. Two years following the removal of definitive hosts, the infection rate decreased by one order of magnitude, while the snails aging less than one year displayed zero infection prevalence. Additionally, phylogenetic analysis of mitochondrial (ND1) and nuclear (ITS1, ITS2) DNA loci revealed negligible intrasite DNA variability of the specimens obtained at the study site in Costa Rica (but not of those obtained earlier in Peru or New Zealand), supporting strongly the hypothesis on focal origin of the infection. The observed dynamics of infection suggests the explanation for the high variability in P. gralli prevalence in intermediate hosts experienced worldwide. We noticed that the reports claiming > 20% prevalence of M. tuberculata infection by P. gralli originated exclusively from foci with known eye infection of the definitive hosts, while the P. gralli infection penetrance < 2% is typically associated with sites, where the infection of definitive hosts was not observed, suggesting that the infected definitive hosts were present onsite only in the past, or were present only at a site upstream or downstream of the respective sampling site. Thus, this is the first evidence on the possible persistence of eye-trematode infection site for over two years following the last confirmed outbreak in its adult hosts.     

Positive phototropism is accelerated in Biomphalaria glabrata snails by infection with Schistosoma mansoni

Parasite-induced behavioral changes in their hosts favor to complete the lifecycle of parasites. Schistosome infection is also known to cause physiological changes in infected freshwater snail intermediate hosts. Here, we report, a novel phenomenon in which Schistosoma mansoni, a highly debilitating worm affecting millions of people worldwide, alters the phototropic behavior of Biomphalaria glabrata, the vector snail. S. mansoni-infection enhanced positive phototropism of vector snails and infected snails spent significantly more time in light. Possibly, these behavioral changes help the parasite to be released efficiently from the infected intermediate hosts, and to infect mammalian hosts.     

Rapid identification of lymnaeid snails and their infection with Fasciola gigantica in Thailand

Freshwater snails of the family Lymnaeidae are the intermediate hosts of the liver fluke Fasciola worldwide. While distinct species have been identified at the molecular level in other parts of the world such data have not been published for Thailand. In this study we collected Lymnaeidae from different localities across Thailand and analyzed their 16S rDNA sequences as a molecular signature for classification. In addition to the ubiquitous Radix rubiginosa, we have confirmed the presence of Austropeplea viridis and Radix swinhoei, for the latter of which the ribosomal rDNA sequences are reported for the first time, in North-Thailand. Based on the obtained 16S rDNA data three primer pairs were designed that allowed rapid identification of these snail species by PCR. To determine their infection status, PCR primers for F.gigantica cathepsin L were used in parallel with the snail 16S rDNA species-specific primers in multiplex PCR analyses. Western blot analysis of total snail protein with a monoclonal anti-F.gigantica cathepsin L antibody confirmed positive cathepsin L PCR results. The developed diagnostic PCR will be of use in risk assessment for transmission of fascioliasis in Thailand.     

Pathology and first report of natural eye infection with the trematode Philophthalmus gralli (Digenea,Philophthalmidae) in Tinamus major (Tinamiformes,Tinamidae), Costa Rica

The eye-fluke Philophthalmus gralli (Philophthalmidae Looss, 1899) was found in six birds known as great tinamous (Tinamus major) reared in a wild animal shelter located in Alajuela, Costa Rica. The birds presented conjunctival hyperemia, blepharitis, anorexia and weakness. Some of them suffered from unilateral blindness and ocular loss. After morphometric analysis, the specimens showed characteristics compatible with the digenean trematode P. gralli. The clinical signs of infection were resolved by manual removal of the adults, treatment with praziquantel and relocation into an environment without a natural water source. In order to determine if an ongoing cycle of this pathogen was present in the shelter, the habitat of the birds was inspected for the presence of infected intermediate hosts and contaminated water and objects. It was found that the snails Melanoides tuberculata acted as the intermediate host, and reared the infectious stages toward other animals, as shown by the reproduction of ocular philophthalmiasis in chickens artificially infected with excysted metacercaria. Moreover, three out of every ten snails found in the place were infected with rediae of P. gralli, raising the possibility of the dispersion of the parasite into new environments as well as the imminent zoonotic risk. The finding of P. gralli in Costa Rica is the first official report in Central America.     

Influence of Trichobilharzia regenti (Digenea: Schistosomatidae) on the Defence Activity of Radix lagotis (Lymnaeidae) Haemocytes

Radix lagotis is an intermediate snail host of the nasal bird schistosome Trichobilharzia regenti. Changes in defence responses in infected snails that might be related to host-parasite compatibility are not known. This study therefore aimed to characterize R. lagotis haemocyte defence mechanisms and determine the extent to which they are modulated by T. regenti. Histological observations of R. lagotis infected with T. regenti revealed that early phases of infection were accompanied by haemocyte accumulation around the developing larvae 2–36 h post exposure (p.e.) to the parasite. At later time points, 44–92 h p.e., no haemocytes were observed around T. regenti. Additionally, microtubular aggregates likely corresponding to phagocytosed ciliary plates of T. regenti miracidia were observed within haemocytes by use of transmission electron microscopy. When the infection was in the patent phase, haemocyte phagocytic activity and hydrogen peroxide production were significantly reduced in infected R. lagotis when compared to uninfected counterparts, whereas haemocyte abundance increased in infected snails. At a molecular level, protein kinase C (PKC) and extracellular-signal regulated kinase (ERK) were found to play an important role in regulating these defence reactions in R. lagotis. Moreover, haemocytes from snails with patent infection displayed lower PKC and ERK activity in cell adhesion assays when compared to those from uninfected snails, which may therefore be related to the reduced defence activities of these cells. These data provide the first integrated insight into the immunobiology of R. lagotis and demonstrate modulation of haemocyte-mediated responses in patent T. regenti infected snails. Given that immunomodulation occurs during patency, interference of snail-host defence by T. regenti might be important for the sustained production and/or release of infective cercariae.