Fasciola hepatica: a study of the shedding of cercariae from Lymnaea truncatula raised under constant conditions of temperature and photoperiod

Investigations on the shedding of cercariae of Fasciola hepatica were carried out in Lymnaea truncatula in order to verify the existence of a low-frequency periodicity in the numerical distribution of metacercariae per snail and per day when the snails are raised under controlled conditions. Preadult L. truncatula were thus collected in the field, individually exposed to two miracidia, and subsequently raised until their death under constant temperature (20 degrees C) and photoperiod (12 h/12 h diurnal rhythm). The 102 snails shedding parasites produced 24,325 metacercariae of which 5% were floating cysts. The daily production of cercariae was maximal during the first 30 days of the patent period, subsequently decreased until day 114, and ceased on day 124. No infradian-type rhythm was noted in the daily distribution of mean values. The snails shed their cercariae in one to 14 waves; 20.6% and 15.7% of the snails produced their parasites in four and five periods respectively. The authors suggest that the seven-day periodicity found by Audousset et al. (1989) in the daily distribution of cercariae produced by three colonies of L. truncatula raised in seminatural conditions must be attributed only to the influence of environmental factors.     

The influence of temperature on the infectivity of Fasciola hepatica miracidia to Lymnaea truncatula

Experiments were performed to study the effect of water temperature on the host-finding capacity (snail localization, attachment, and penetration) of Fasciola hepatica miracidia. Specimens of Lymnaea truncatula were exposed to miracidia labeled in vivo with radioselenium, and the radioactivity which subsequently was confined to the snails was taken as a measure of the host-finding capacity of the parasite. The minimum temperature required for host-finding was 5 to 6 C and the optimum temperature was in the range between 15 and 26 C. The lack of host-finding capacity below 5 to 6 C could be reversed experimentally by raising the temperature. A clear inverse relationship was demonstrated between the environnd 24 C the host-finding capacity ceased after 24 to 30, 20 to 24, and 13 to 20 hr, respectively.     

Unusual transmission of the liver fluke, Fasciola hepatica, by Lymnaea glabra or Planorbis leucostoma in France

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.     

Outcome and predictive factors in children with chronic idiopathic musculoskeletal pain

The 18S rDNA sequences of the six most common European Lymnaeidae species (Mollusca:Gastropoda:Basommatophora) have been obtained by direct PCR cycle sequencing and silver staining methods. The sequence alignment and secondary structures of the 18S rRNA gene of Lymnaea stagnalis, L. auricularia, L. peregra, L. palustris, L. glabra, and L. truncatula are analyzed. This gene proves to be a good marker for both specific determination and supraspecific lymnaeid phylogeny. The malacological importance is evident, considering the specific determination problems of individual snails and the present systematic chaos in Lymnaeidae due to their pronounced morphoanatomic uniformity, which makes a classification by traditional methods impossible. The majority (17) of the total of 43 nucleotide-substituted positions appears to be confined to a small region included in helix E10-1 of the variable region V2, enabling species group distinction: (1) the first sequence is common to L. auricularia and L. peregra; (2) the second sequence is unique to L. truncatula; and (3) the third sequence is identical for L. glabra, L. palustris, and L. stagnalis. The other 26 nucleotide-substituted positions are dispersed over the entire gene, although four grouped nucleotide positions in helix 6 of V1 are of interest in distinguishing L. glabra from both L. palustris and L. stagnalis. The phylogenetic trees obtained by comparison with four other molluscan species (a polyplacophoran, two bivalves, and a stylommatophoran gastropod) show the presence of four well-defined subgenera among the genus Lymnaea sensu lato: (1) Lymnaea (Radix), (2) Lymnaea (Galba), (3) Lymnaea (Leptolimnaea), and (4) Lymnaea (Lymnaea). Two branches, L. auricularia-L. peregra-L. truncatula and L. glabra-L. palustris-L. stagnalis, are worth mentioning from the parasitological point of view, since the two digenean species of large medical and veterinary impact transmitted by lymnaeids, Fasciola hepatica and F. gigantica, appear to be linked to the first branch.     

Host choice by larval parasites: a study of Biomphalaria glabrata snails and Schistosoma mansoni miracidia related to host size

Within snail/trematode associations the age/size of the host at infection has consequences with regard to miracidial infection success, further intramolluscan parasite development and reproduction, and the host response, mainly in terms of growth and reproductive effort. Taking into account these differences, we were interested in determining whether miracidia could discriminate and make a choice between snails of different sizes. Using the Schistosoma mansoni/Biomphalaria glabrata system, we compared data on the snail infection rate and the mother sporocyst abundance among three size classes of snails (juvenile, subadult, and adult) exposed separately or together to the parasite larvae. When exposed individually, juvenile snails (3-5 mm) had significantly higher prevalence and abundance values than did subadult snails, followed by adult snails. In contrast, when snails of the three size classes were exposed together in heterogeneous size groups the prevalence and abundance values were always significantly higher for subadult snails of the 7- to 9-mm class than for juvenile and adult snails. A host choice experiment confirmed that significantly more miracidia were attracted by subadult snails, suggesting that the parasite has been selected for specific locating and recognition mechanisms increasing the infection rate of subadult snails when the latter have been exposed in a heterogeneous size group. Selective forces that may be responsible for such a preferential infectivity of the parasite vis-à-vis particular host age/size class are discussed in relation to host resources and host responses.     

Miracidial infectivity of Hypoderaeum conoideum (Trematoda: Echinostomatidae): differential susceptibility of two lymnaeid species

A study was made of the infectivity of Hypoderaeum conoideum miracidia to a range of laboratory-reared specimens of freshwater snail species (Lymnaea peregra, L. corvus, Physella acuta, and Gyraulus chinensis) that coexist with the parasite in the same natural habitat. L. peregra and L. corvus were found to be equally susceptible to the parasite when specimens of each snail species were singly exposed to miracidia. However, when miracidia could choose either lymnaeid species, they showed a high degree of specificity toward L. peregra. The results obtained suggest that H. conoideum miracidia are capable of distinguishing among these lymnaeids in their orientation to the host. This indicates that miracidia might achieve specificity before actually contacting the snail host and suggests that during the host-snail orientation process they respond to signals different from those generated upon snail contact and invasion. The specificity toward L. peregra observed in H. conoideum miracidia seems to indicate adaptation to the snail community in their natural habitat, resulting in enhancement of their transmission.     

Intraarterial infusion of anti-cancer agents. (A suitable technic introducing a polyethylene tube, with angiographic visualization, through the deep femoral artery directly into the artery in the abdomen selected for injection, using two kinds of guidewires) (author''s transl)

The authors have infected 127 B. glabrata by 2 miracidia of S. mansoni, either on the Same day, or introducing a second miracidium after 3, 7 and 16 days. 1 All the groups of planorbid snails had a low percentage of positivity (31 to 41 %). 2 The first cercarial emissions, in the 4 groups were scattered in the time, the delay being in correlation with the second infection. 3 In the snails reinfected after 3 and 7 days occured the highest emissions, those with the simultaneous double infection or second infection delayed to 16 days, had the lesser emissions. 4 An interval cycle of about 3 weeks was discovered for the highest emissions. All those phenomena are probably due to competition between the sporocysts born from both miracidia. Moreover, evident reduction of the fecundity in the positive snails were shown only after the beginning of the cercarial emissions, while a normal, or even increased fecundity was established in the prepatent period of the infected snails.     

Selective interference with granulocyte function induced by Echinostoma paraensei (Trematoda) larvae in Biomphalaria glabrata (Mollusca)

Various trematode larvae can interfere with the host snail's resistance to the same or unrelated trematode species, chiefly, it appears by interference with the function of the host's granulocytes. In Biomphalaria glabrata infected with the trematodes, Echinostoma paraensei, granulocytes lose their ability to encapsulate the larvae of trematodes to which the hosts were previously resistant. However, the granulocytes in these snails retain their ability to encapsulate injected latex spheres, or larvae of the metastrongyle nematode, Angiostrongylus malaysiensis, and to phagocytose epidermal plates cast off by miracidia of the trematode, Schistosoma mansoni. Cellular infiltration in injured preputial tissue of the snail also was not suppressed by the presence of E. paraensei larvae. Interference with the granulocyte function in B. glabrata induced by E. paraensei infection therefore appears to be a selective phenomenon.     

Bilateral lower extremity evaluation of deep venous thrombosis with color flow and compression sonography

Fasciola hepatica miracidia were experimentally introduced into five sites colonized by Lymnaea palustris over a period of 4 or 6 years. In the first four ponds, a progressive increase in the prevalence of the spring-generation juvenile snails was observed (from 0.4 to 18.1%), with a corresponding increase in the shell height of infected snails (from 3.6 to 7.8 mm). In the fifth habitat, the pond dried in 1990 causing the prevalence to drop as compared with the initial values and to subsequently increase in 1991-1992.     

1H NMR study of dynamics and thermodynamics of acid-alkaline transition in ferric hemoglobin of a midge larva (Tokunagayusurika akamusi)

Molluscicidal activity of the herebicides 2,4-D and Graminol, as well as both extracts and dry powder of the plant Azolla pinnata were evaluated against B. alexandrina snails. It was observed that 2,4-D proved to be the most toxic compound among he tested ones, showing LC90 of 52 ppm after 24 h of exposure. Ethanol extract of Azolla showed the highest molluscicidal activity against the tested snails compared with the other extracts and dry powder (LC90 = 3300 ppm). Ethanol extract at 6600 ppm after 3 h of exposure killed 100% and 19.4% of S. mansoni miracidia and cercariae, respectively. The molluscicidal activity of 2,4-D was not influenced by the presence of Azolla (900 plants/liter) for 7 days, while Graminol effect was significantly reduced. However, the infectivity of S. mansoni miracidia to B. alexandrina snails was not affected by Azolla existence.     

Reducing perineal trauma: implications of flexion and extension of the fetal head during birth

Among the determinant factors in the resistance and susceptibility of Biomphalaria to Schistosoma mansoni, hemocytes play an important role. Aiming at studying S. mansoni/Biomphalaria interactions related to hemocytes, the first step is certainly connected with the standardization of this cell population in uninfected Biomphalaria. In this way, quantification of this cell population in hemolymph, as well as its phagocitary capacity, have been determined for the first time. Furthermore, using susceptible and resistant strains of B. glabrata and B. tenagophila, the hemocytegram and phagocytary capacity of hemocytes after infection with S. mansoni were determined too. Resistant and susceptible strains of B. glabrata (BA and BH, respectively), as well as resistant and susceptible strains of B. tenagophila (TAIM and CF, respectively) were infected with 10 miracidia of the LE and SJ strains of S. mansoni, respectively. These infected snails and respective uninfected controls were assessed in relation to the number of circulating hemocytes and alteration in the phagocytary capacity, by using Zymozan and MTT. Reading was taken by means of a spectrophotometer at 5 hours and 1, 2, 5, 10, 20 and 30 days after infection. The results showed a decrease in population of the circulating phagocytary cells, 5 hours after infection. One day post-infection, the circulating cells of the susceptible snails showed an increased metabolic activity, but the same event could not be observed in the resistant strains. In the subsequent observation periods, significant differences among the strains studied could not be observed until the end of the experiment.     

Effects of snail-conditioned water from Biomphalaria glabrata on hatching of Echinostoma caproni miracidia

In vitro studies were done on Echinostoma caproni eggs with fully developed miracidia to determine the effects of snail-conditioned water (SCW) from Biomphalaria glabrata on miracidial hatching in the light. Observations were made on miracidial hatching at 4, 8, 12, 16, 20, and 24 h in multiwell chambers in the presence of SCW (experimentals) versus controls in artificial spring water (ASW). The number of hatched eggs was significantly greater (Student's t-test, P<0.05) in SCW at all times as compared with those maintained in ASW. Significantly greater hatching was also obtained when snails were maintained in intact or perforated dialysis sacs in multiwell chambers as compared with sacs without snails. Agar plugs impregnated with SCW or the hydrophilic fraction of SCW that had been extracted in chloroform-methanol (2:1) did not influence significant hatching. However, the lipophilic fraction of the SCW extract caused significant hatching. Substances in SCW significantly increase hatching of E. caproni miracidia, but details on what these compounds are remain obscure.     

Developmental plasticity of respiratory behavior in Lymnaea

The authors investigated the contribution of experience to development and maintenance of pulmonary respiration in Lymnaea stagnalis. Respiration in L. stagnalis is bimodal via both the skin and the lung. Rearing snails from eggs to adulthood while preventing lung respiration (differentially reared snails) showed that L. stagnalis can develop and survive without pulmonary respiration. These snails were able to open and close their pneumostome when given the opportunity as adults. However, quantitative aspects of their respiratory behavior were significantly altered. Prevention of pulmonary respiration in adult, normally reared snails also induced behavioral changes. Comparison of these changes with those in differentially reared snails revealed specific developmental effects, which were reversible. Thus, this is a suitable model system for studying questions related to behavioral plasticity.     

First report of the isolation of an adult worm of the genus Brachylaima (Digenea: Brachylaimidae), from the gastrointestinal tract of a human

A 78-year-old woman presented with an 18-month history of intermittent diarrhoea. Examination of her stools revealed brachylaimid eggs, which were present in three separate specimens over a week. After treatment with praziquantel a degenerate adult Brachylaima species was recovered from her faeces. She lived in a rural area of South Australia and ate vegetables grown in her own garden which had been infested with helicid snails. In south Australia these introduced European helicid snails are commonly infected with brachylaimid intermediate larval stages and are considered to be the source of the human infection.     

Seasonal variation in molluscicidal activity of Solanum nigrum L

Solanum nigrum L. leaves and fruits were shown to have molluscicidal activities against snails transmitting schistosomiasis and fascioliasis. In the present study, their molluscicidal activity against adult Biomphalaria alexandrina snails was assessed to determine whether plants collected at various seasons would have different degrees of toxicity. Leaves and fruits of three S. nigrum varieties were collected from Faiyoum and/or Giza during the four seasons. Leaves collected in autumn had the highest effect (LC50-35.4) followed by spring (LC50 = 44.36), summer (LC50 = 46.7) and winter (LC50 = 100.4). Toxicity of plant extracts was also affected by other seasonal dependent factors. These are the duration of plant exposure to direct sunlight and the size of the fruits. S. nigrum (black fruits) was more toxic (LC50 = 18.1) than the other two types, S. nigrum v. vellosum (yellow fruits) (LC50 = 38.9) and S. nigrum v. juidaicum (red fruits) (LC50 = 34.7).     

Susceptibility to chemotherapeutic agents of Schistosoma mansoni isolates from patients treated with oxamniquine and praziquantel and not cured

Ten inhabitants of Itaquara, Bahia, Brazil treated with oxamniquine and subsequently praziquantel were not cured. Schistosoma mansoni isolates derived from these patients were studied. Snails were infected with miracidia derived from the feces of these patients and the cercariae produced used to infect albino mice. The animals were then treated with a single oral dose of oxamniquine (25, 50 and 100mg/kg) or praziquantel (100, 200 and 400 mg/kg). The response to chemotherapy was significantly different in some of the isolates although it was not possible to characterize any of them as resistant. In addition, DNA analysis of the isolates by means of "Random Amplified Polymorphic DNA" indicated a low degree of variability as compared with a laboratory strain, LE. Thus, it was not possible to characterize these organisms at a genetic level as a distinct strain.     

Phosphatase-negative mutants of Legionella pneumophila and their behavior in mammalian cell infection

Microbial phosphatases are known or suspected to play a role in the pathogenesis of several intracellular pathogens, including Legionella micdadei. Legionella pneumophila also possess phosphatase activities, but their possible roles in cellular infection are unknown. We generated mutants of a serogroup 1 isolate of L. pneumophila that lack the major phosphatase. Isolation of a Pho- mutant after random mutagenesis with transposon MudII4041 allowed us to dissociate the major alkaline phosphatase (pH optimum approximately 8) from a minor acid phosphatase activity. Both activities were concentrated in the bacterial periplasm. The gene encoding the major alkaline phosphatase (pho) was cloned by expression in E. coli and used to generate a site directed mutation in two L. pneumophila strains. Each parent-mutant pair was compared in a U937 cell tissue culture assay for capacity to infect, lyse, and grow within mammalian cells. Although the parental stains differed in their U937 cell cytopathicity, neither was significantly more infective than its Pho- derivative, suggesting that the alkaline phosphatase activity is not essential for cellular infection. Because they are not attenuated, Pho- mutants can be used to generate gene fusions with E. coli alkaline phosphatase to study and secretion and cellular infectivity in L. pneumophila.     

Pre-B cell receptor-mediated selection of pre-B cells synthesizing functional mu heavy chains

Ig gene rearrangements could generate V(H)-D-J(H) joining sequences that interfere with the correct folding of a mu-chain, and thus, its capability to pair with IgL chains. Surrogate light (SL) chain might be the ideal molecule to test the capacity of a mu-chain to pair with a L chain early in development, in that only pre-B cells that assemble a membrane mu-SL complex would be permitted to expand and further differentiate. We have previously identified two SL chain nonpairing V(H)81X-mu-chains with distinct V(H)-D-J(H) joining regions. Here, we show that one of these V(H)81X-mu-chains does not rescue B cell development in J(H) knock-out mice, because flow cytometric analysis of bone marrow cells from V(H)81X-mu transgenic J(H) knock-out mice revealed normal numbers of pro-B cells, but essentially no pre-B and surface IgM+ B cells. Immunoprecipitation analysis of transfected pre-B and hybridoma lines revealed that the same mu-chain fails to pair not only with SL chain but also with four distinct kappa L chains. These findings demonstrate that early pre-B cells are selected for maturation on the basis of the structure of a mu-chain, in particular its V(H)-D-J(H) joining or CDR3 sequence, and that one mechanism for this selection is the capacity of a mu-chain to assemble with SL chain. Therefore, we propose a new function of SL chain in early B cell development: SL chain is part of a quality control mechanism that tests a mu-chain for its ability to pair with conventional L chains.     

Helminths of six species of Anolis lizards (Polychrotidae) from Hispaniola, West Indies

The gastrointestinal tracts of gekkonid lizards from Guam (Gehyra mutilata [n = 4], Gehyra oceanica [n = 11], Hemidactylus frenatus [n = 43], and Lepidodactylus lugubris [n = 38]) and Rota (Gehyra oceanica [n = 2], Hemidactylus frenatus [n = 13], and Lepidodactylus lugubris [N = 20] were examined for helminths. Found were 2 species of cestodes, Cylindrotaenia allisonae, Oochoristica javaensis, 1 species of trematode, Allopharynx macallisteri, and 5 species of nematodes, Pharyngodon lepidodactylus, Spauligodon gehyrae, Spauligodon hemidactylus, Skrjabinelazia machidai, Oswaldocruzia sp. New host records included Gehyra mutilata for Spauligodon hemidactylus, Gehyra oceanica for Oochoristica javaensis, Hemidactylus frenatus for Skrjabinelazia machidai, and Lepidodactylus lugubris for Cylindrotaenia allisonae and Oswaldocruzia sp. These helminths are known only from Pacific Islands and the Australian biogeographic realm.     

Interspecies aminopeptidase-N chimeras reveal species-specific receptor recognition by canine coronavirus, feline infectious peritonitis virus, and transmissible gastroenteritis virus

We report that cells refractory to canine coronavirus (CCV) and feline infectious peritonitis virus (FIPV) became susceptible when transfected with a chimeric aminopeptidase-N (APN) cDNA containing a canine domain between residues 643 and 841. This finding shows that APN recognition by these viruses is species related and associated with this C-terminal domain. The human/canine APN chimera was also able to confer susceptibility to the porcine transmissible gastroenteritis virus (TGEV), whereas its human/porcine homolog failed to confer susceptibility to CCV and FIPV. A good correlation was observed between the capacity of CCV, FIPV, and TGEV to recognize the different interspecies APN chimeras and their ability to infect cells derived from the relevant species. As an exception, TGEV was found to use a human/bovine APN chimera as a receptor although itself unable to replicate in bovine cells.