In vivo efficacy of praziquantel against Echinoparyphium aconiatum (Trematoda: Echinostomatidae) parasitizing the great pond snails Lymnaea stagnalis (Gastropoda: Lymnaeidae)

The present study had a practical goal. I aimed to determine whether praziquantel could reduce the production of Echinoparyphium aconiatum (Trematoda: Echinostomatidae) cercariae in infected snails Lymnaea stagnalis (Gastropoda: Lymnaeidae) without killing the hosts. Praziquantel is a broad-spectrum antihelminth agent. It caused a total cessation of cercaria shedding when the praziquantel concentration in the treatment bath was 10 mg/L and the treatment time was 30 h or longer. A next research step which has to be taken before giving detailed recommendations about using praziquantel for ceasing production of E. aconiatum cercariae in parasitized snails is to follow the survivorship and performance of treated snails after a praziquantel exposure for longer than in this medium-term (3 days) experiment.     

Relative importance of chemical attractiveness to parasites for susceptibility to trematode infection

While the host immune system is often considered the most important physiological mechanism against parasites, precontact mechanisms determining exposure to parasites may also affect infection dynamics. For instance, chemical cues released by hosts can attract parasite transmission stages. We used the freshwater snail Lymnaea stagnalis and its trematode parasite Echinoparyphium aconiatum to examine the role of host chemical attractiveness, physiological condition, and immune function in determining its susceptibility to infection. We assessed host attractiveness through parasite chemo‐orientation behavior; physiological condition through host body size, food consumption, and respiration rate; and immune function through two immune parameters (phenoloxidase‐like and antibacterial activity of hemolymph) at an individual level. We found that, although snails showed high variation in chemical attractiveness to E. aconiatum cercariae, this did not determine their overall susceptibility to infection. This was because large body size increased attractiveness, but also increased metabolic activity that reduced overall susceptibility. High metabolic rate indicates fast physiological processes, including immune activity. The examined immune traits, however, showed no association with susceptibility to infection. Our results indicate that postcontact mechanisms were more likely to determine snail susceptibility to infection than variation in attractiveness to parasites. These may include localized immune responses in the target tissue of the parasite. The lack of a relationship between food consumption and attractiveness to parasites contradicts earlier findings that show food deprivation reducing snail attractiveness. This suggests that, although variation in resource level over space and time can alter infection dynamics, variation in chemical attractiveness may not contribute to parasite‐induced fitness variation within populations when individuals experience similar environmental conditions.     

The Early Worm Catches the Bird? Productivity and Patterns of Trichobilharzia szidati Cercarial Emission from Lymnaea stagnalis

Digenean trematodes are common and abundant in aquatic habitats and their free-living larvae, the cercariae, have recently been recognized as important components of ecosystems in terms of comprising a significant proportion of biomass and in having a potentially strong influence on food web dynamics. One strategy to enhance their transmission success is to produce high numbers of cercariae which are available during the activity peak of the next host. In laboratory experiments with 13 Lymnaea stagnalis snails infected with Trichobilharzia szidati the average daily emergence rate per snail was determined as 2,621 cercariae, with a maximum of 29,560. During a snail’s lifetime this summed up to a mass equivalent of or even exceeding the snail’s own body mass. Extrapolated for the eutrophic pond where the snails were collected, annual T. szidati biomass may reach 4.65 tons, a value equivalent to a large Asian elephant. Emission peaks were observed after the onset of illumination, indicating emission synchronizing with the high morning activities of the definitive hosts, ducks. However, high cercarial emission is possible throughout the day under favorable lightning conditions. Therefore, although bird schistosomes, such as T. szidati constitute only a fraction of the diverse trematode communities in the studied aquatic ecosystem, their cercariae can still pose a considerable risk for humans of getting cercarial dermatitis (swimmer''s itch) due to the high number of cercariae emitted from infected snails.     

Echinostome rediae: effect of pretreatment with cold on transplantation to uninfected snails.

When donor snails, Lymnaea stagnalis, infected with Echinoparyphium aconiatum were held at 13 C for at least 24 days, transplantation of newborn rediae from them to uninfected snails was 2.5 times as successful as when donor snails were held at 23 C. The difference does not seem to be do to the fact that rediae transferred from 13 C-snails were slightly larger than those from 23 C. This effect of temperature was not observed in analogous experiments with another echinostome, Isthmiophora melis. The mechanism of this phenomenon found in the experiments with E. aconiatum is still unknown.     

Trematode larvae in snails of Lake Glubokoe

In the Augusts of 1984 and 1985, 6 species of snails in Lake Glubokoe were investigated: Viviparus viviparus (Linne, 1758), Lymnaea ovata (Draparnaud, 1805), L. stagnalis (Linne, 1758), L. corvus (Gmelin, 1791), Planorbis planorbis (Linne, 1758), and Planorbarius corneus (Linne, 1758), which were found to be infected by 9 species of trematode larvae. V. viviparus showed the highest rate of infection — 48.15% ± 6.8. The larvae of trematodes belonging to the families Echinostomatidae and Plagiorchidae predominate in the snails of the lake.     

Individual and population variation in cercariae of bird schistosomes of the <Emphasis Type="Italic">Trichobilharzia ocellata</Emphasis> species group as revealed with the polymerase chain reaction

The polymerase chain reaction with arbitrary (RAPD-PCR) or specific primers was used to study the population variation and to identify the species in cercariae of schistosomes of the Trichobilharzia ocellata species group (Trematoda, Schistosomatidae). In total, 28 cercariae were obtained from two spontaneously invaded mollusks Lymnaea stagnalis (LS) and L. ovata (LO), which were collected in different ponds of Moscow. RAPD-PCR was carried out with two arbitrary primers, OPA9 and OPB11, which each detected different levels of individual and among-group variation and revealed considerable genetic differentiation of cercariae from different host mollusks. To check whether the cercariae of the two samples belong to one species, sequencing was performed with a region corresponding to intergenic transcribed spacer 2 (ITS2), which was earlier proposed for cercaria identification in three European species of bird schistosomes of the genus Trichobilharzia (T. franki, T. regenti, and T. szidati). The ITS2 sequences of two LO cercariae were identical, each consisted of 319 bp, and showed 100% homology to the T. franki ITS2 sequence. The ITS2 sequences of two LS cercariae were identical, each consisted of 323 bp, and showed 99.4% homology to the T. szidati counterpart. The causes of genetic variation in cercariae and prospects of using RAPD markers to study different stages of the life cycle in trematodes are discussed.Translated from Genetika, Vol. 41, No. 1, 2005, pp. 17–22.Original Russian Text Copyright © 2005 by Semyenova, Chrisanfova, Filippova, Beer, Voronin, Ryskov.     

Relationship between prevalence of trematode parasite <Emphasis Type="Italic">Diplostomum</Emphasis> sp. and population density of its snail host <Emphasis Type="Italic">Lymnaea stagnalis</Emphasis> in lakes and ponds in Finland

Diplostomum sp. is a trematode parasite that infects aquatic snails, e.g. Lymnaea stagnalis (Gastropoda: Lymnaeidae), fish, and fish-eating birds. Ponds and lakes (n = 28) located in Finland between latitudes 61°45′N and 65°30′N were sampled for L. stagnalis, the first intermediate host for Diplostomum sp. L. stagnalis were found in 22 sites out of 28, and Diplostomum sp. in 10 of the 22 snail populations. Among the L. stagnalis populations that were infected by Diplostomum sp., the mean prevalence was 12.8%. Diplostomum sp. occurred in only one out of the seven L. stagnalis populations in four large lakes, but in 9 out of the 16 L. stagnalis populations in small lakes. In the pooled data, a positive correlation (r _s = 0.427; P = 0.047; n = 22) between L. stagnalis density and Diplostomum sp. prevalence was found. The results suggest that Diplostomum sp. is fairly common in L. stagnalis populations in small lakes but rare in large lakes. Furthermore, although trematode parasites, in general, should have a negative effect on snail population density, the study indicates that the relationship between host density and parasite prevalence may greatly differ for individual trematode species, such as Diplostomum sp.     

Black Sea mollusc, Cerithium vulgatum (Gastropoda, Cerithiidae), a new intermediate trematode host

The mollusc Cerithium vulgatum was first recorded as a new intermediate host of trematodes from the Black Sea. Two species of cercariae (Cercaria I and Ceracaria II) parasitizing this mollusc were found and described. The general infestation of molluscs was 7.54 per cent. Eight species of larval trematodes parasitzing 7 other species of sea molluscs were found.     

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.     

Neuromodulatory effects of hydrogen peroxide on central neurons in the feeding network of the mollusc <Emphasis Type="Italic">Lymnaea stagnalis</Emphasis>

Hydrogen peroxide at a concentration of 100 μM was found to exert a pronounced modulatory effect on motor (R/L cells in B1–B4 clusters) and modulatory (R/L cerebral giant cells) neurons in the feeding neural network of the mollusc Lymnaea stagnalis as manifested in changes in the firing rate, membrane potential level and spike amplitude in these cells. The observed effects were reversible, transient, and reached their peak values in 1 min since application of the preparation. Injection of hydrogen peroxide into the cavity of the cephalopedal sinus resulted in no statistically significant changes in the parameters of mollusc feeding behavior. Hydrogen peroxide is assumed to act as a rapid neuromodulator towards neurons of the central feeding rhythm generator in Lymnaea stagnalis.     

State of antioxidative protection in central nervous ganglia of the mollusc <Emphasis Type="Italic">Lymnaea stagnalis</Emphasis> at modulation of activity of the NO-ergic system

In experiments on mollusc Lymnaea stagnalis, the state of antioxidative protection is studied in central nervous ganglia during a long-term activation (inhibition) of synthesis of nitrogen monoxide (NO) in the body. The effect of the blocker of NO-synthase N^G-nitro-L-arginine (L-NNA) at the background of enhancement of pulmonary respiration has been found to be associated with a rise of levels of reduced glutathione and TBK-active products in the nervous tissue at preservation of a relatively high superoxide dismutase activity and a low glutathione peroxidase activity compared with the control group and the animals treated with the metabolic precursor of NO synthesis L-arginine. In spite of the revealed disturbances of balance of the body proand antioxidative system, DNA electrophoresis detected no products of its degradation, which can indicate the absence of massive programmed death of the nervous tissue cells in Lymnaea stagnalis during modulation of activity of the NO-ergic system.     

Diversity of trematode larvae in gastropod molluscs in water bodies of Belarus

The analysis of literary data and own investigations of gastropod molluscs on existence of cercariae of trematodes in Belarus was carried out. 13 856 specimens of 11 species and 2 species complexes of freshwater gastropod molluscs were examined on infection with cercariae of trematodes in 2010 by the authors of this article. Total infestation was 13.1% and infection of some molluscs species varied from 0.4% (Dnieper River, Gomel region) to 66.7% (Lukomskoe reservoir, Vitebsk region). The cercariae of 90 trematode species were found. The majority (51 species) at the adult stage parasitize birds. The cercariae of 14 trematode species have medical significance.     

Isthmiophora melis (Trematoda,Echinostomatidae): Normal adults after 41 snail passages in 7 years of redial transplantation

A strain of Isthmiophora melis rediae has been kept for 8 years by transplantation from infected to uninfected snails. This strain has passed with undiminished redial reproductive ability in 45 transplantation steps through 46 Lymnaea stagnalis specimens which corresponds to a minimum of 93 successive redial generations. It was possible to start a normal sexual cycle of the trematode with cercariae deriving from rediae of the 41st snail passage, corresponding at least to the 83rd redial generation. Generative adults and infective miracidia were obtained. Hence the sexual reproduction potentiality of Isthmiophora melis has not been impaired by this artificial extension of the intramolluscan development for more than 7 years.     

Assessment of the joint effect of thermal stress,pollution, and parasitic infestation on the activity of antioxidative enzymes in pulmonate mollusk <Emphasis Type="Italic">Lymnaea stagnalis</Emphasis>

This study makes a multifactor analysis of the joint effect of reservoir contamination, parasitic infestation, and experimental hyperthermia on the activity of antioxidative enzymes (peroxidase, catalase, and glutathione S-transferase) in the pulmonate mollusk Lymnaea stagnalis from the clean Naroch Lake and contaminated Chizhovsky Reservoir in the Republic of Belarus. The activity of peroxidase is ascertained to be affected most strongly by the simultaneous effect of parasitic infestation and reservoir contamination and parasitic infestation coupled with experimental hyperthermia. The joint effect of parasitic infestation and reservoir pollution is found to influence the activity of catalase. Meanwhile, no correlation is found between the studied factors and the activity of glutathione S-transferase. It is shown that there is no mutual influence of the three studied factors on the activity of the investigated enzymes.     

Coordination of locomotor activity of mollusc Lymnaea stagnalis at nutrition: Role of the internal medium acid-base balance (pH)

The correlation was studied between the hemolymph pH value and the locomotion rate in the mollusc Lymnaea stagnalis at different degree of activation of the food-procuring behavior. It is established that the hemolymph pH value remains unchanged in the satiated (hungry), freely moving Lymnaea individuals, and, hence, it does not determine the locomotor activity of molluscs in these functional states. In the course of food consumption, locomotion decreases significantly, which is due to an observed decrease of the hemolymph pH. This effect is realized through the corresponding changes of electrical activity of motoneurons of the locomotor network of the Lymnaea nervous system (PeA-cluster). It is suggested that the pH value plays the modulating role in coordination of activities of the nervous centers of the mollusc nutrition and locomotion.     

Laboratory Studies of Parasite Transmission Aspects between Hydrobia spp. (Gastropoda) and Corophium volutator (Amphipoda)

Aspects of parasite transmission between Hydrobia spp. and Corophium volutator, first and second intermediate host of digenetic trematodes, were investigated under laboratory conditions. H. ventrosa is used as an intermediate host by several trematode species. Under laboratory conditions the most frequently observed emergence from H. ventrosa was of cercariae of Maritrema subdolum. The number of cercariae shed per day varied considerably. It was observed that 30 cercariae on average and up to 450 cercariae at maximum can emerge from a single H. ventrosa per day. Cercarial production continued until the death of the snails. The life-span of cercariae of the species M. subdolum decreased as the water temperature increased. It can be concluded that under natural conditions the cercariae, after emerging, have a maximum period of 1 day in which to seek out their second intermediate host C. volutator. Almost all specimens of C. volutator exposed to cercariae of the species M. subdolum died within the test period of 6 days. High average cercarial densities caused short life-spans (<50 h), while at lower densities longer survival times were possible. For cercariae of other Trematoda species, we were unable to find any equally clear evidence of a reduction in the survival rate of C. volutator within the test period. Mortality of C volutator, and other effects of infestation, as observed in our experiment, can be assumed to be a result of the penetration process of the cercariae but they also can be attributed to the mesocercariae.     

A histoenzymatic investigation of galactogen synthesis in the albumen gland of Helix pomatia and Lymnaea stagnalis

Summary A histoenzymatic investigation of Galaotogen synthesis has been attempted in the albumen gland of Helix pomatia and Lymnaea stagnalis. Using histoenzymatic methods the authors succeeded in finding only two enzymes (E.C. 2.7.7.9.=uridylyltransferase and E. C. 5.1.3.2.=epimerase) demonstrated by biochemical assays and isolated. They therefore assume that there may be other metabolic pathways in addition to those demonstrated by biochemical methods.     

Parthenites and cercaria of trematodes in the snails Lymnaea saridalensis (Gastropoda, Pulmonata), which inhabits the watershed area of Chany Lake (the South of Western Siberia)

The full list of parthenogenetic larvae and cercaria is presented for the first time for the freshwater snail L. saridalensis (Gastropoda, Pulmonata), which inhabits the watershed area of Chany Lake. It was found that this snail species plays a significant role as the first intermediate host in the life cycles of trematodes in the southern part of Western Siberia. The invasion parameters were calculated for different parasite species. It was found that 50% of the L. saridalensis population serves as the first intermediate host for eleven trematode species that belong to six families: Plagiorchiidae, Echinostomatidae, Diplostomatidae, Strigeidae, Notocotylidae, and Schistosomatidae. The prevalence of infection by each trematoda species in L. saridalensis was determined. Five species of trematodes from the Plagiorchiidae and Echinostomatidae families formed the center of the parasite community, viz., the species Plagiorchis elegans, P. mutationis, Opisthioglyphe ranae, Molinella anceps, and Echinoparyphium aconiatum. Two species, Plagiorchis mutationis and P. multiglandularis, were recorded in the studied area (the Chany Lake watershed area) for the first time at the stages of parthenite and cercaria. It was also found that 1% of L. saridalensis population have multiple (mostly two-species) infections.     

Acid-Base Balance Modulates Respiratory and Alimentary Behavior of the Mollusc Lymnaea stagnalis

An active reaction (change of pH) in hemolymph of the freshwater pulmonate mollusc Lymnaea stagnalis is studied in various physiological states: hunger, satiety, and alimentary excitation. In satiated animals the hemolymph pH is shifted to the acid area (7.6 ± 0.003) as compared with hungry animals (7.9 ± 0.05). The satiated individuals are characterized by an increased respiratory activity and decreased food consumption as compared with hungry animals. Acidification of the medium leads to an increase of excitability of the respiratory network interneurons (RPeD1, VD4, VD1/RPD2) and to inhibition of activity of the alimentary network interneurons (R/L CGC), whereas alkalization, to opposite effects. It is suggested that pH is one of factors coordinating activity of functional systems of Lymnaea stagnalis due to effect on neuronal correlates of the respiratory and alimentary behavior.     

Respiratory behavior in the pond snail Lymnaea stagnalis

Summary This study describes the neural basis of respiratory behavior in a pulmonate mollusc, Lymnaea stagnalis. We describe and identify muscles of the respiratory orifice (pneumostome) and mantle cavity as well as relevant motor neurons innervating these muscles. All of these identified motor neurons are active during spontaneously occurring respiratory behavior and a sporadically occurring synaptic input, termed Input 3, controls the activities of these motor neurons. This spontaneous input can also be recorded from isolated brain preparations, suggesting that the respiratory motor program is generated centrally. However, evidence is also presented that in semi-intact preparations the role of peripheral feedback is important for the initiation and termination of respiratory behavior in Lymnaea.