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Schistosome infection rates in field snails: Schistosoma mansoni in Biomphalaria pfeifferi from Kenya.

Six collections of Biomphalaria pfeifferi were made in the Nairobi and Machakos areas of Kenya. Individual cercarial shedding (Method A) showed that 6.9% (65/938) of the snails had mature 'patent' Schistosoma mansoni infections (range 3.1--34% in the six collections). Crushing (Method B) about one-third of the remaining snails detected no additional mature infections but yielded 1.4% (four snails from one collection) with secondary sporocysts from immature 'prepatent' infections. Weekly shedding observations for five weeks (Method C) on the remainder of the snails showed that 6.9% (40 snails from four collections--range 1.5--30%) shed cercariae maturing from infections immature at the time of collection. As Method B detected no cercariae, Method A apparently detected all 'patent' infections reliably, despite the stress suffered by the snails before examination. Method C was simpler and more efficient than Method B in the detection of infections which were immature at the time of collection. Judging from the rate at which infections matured in Method C, field snails in heavily endemic areas are subjected to pulses of infection rather than to a continuous flow of miracidia. The 'patent' infection rate was not correlated to the 'prepatent' infection rate. Some of the implications of these findings to mathematical models of schistosome transmission are discussed.

Animals↗

Tissue reactions induced by Schistosoma mansoni in Biomphalaria glabrata.

In Biomphalaria glabrata with a strong natural resistance, Schistosoma mansoni sporocysts are rapidly encapsulated by granulocytes and killed, mainly by the strong phagocytic activity of the cells. Irradiated Echinostoma paraensei sporocysts seem able to suppress the function of the granulocytes. Tissue reactions in snails with self-cure demonstrate: involvement of two types of cells, granulocytes and hyalinocyte-like cells; formation of amoeba-fibrous capsules; limited tendency of granulocytes to become attracted to the parasites; a slow process of parasite destruction; and a possible involvement of humoral factors. It seems that there is partial suppression of the granulocyte function in smails with self-cure.

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Displacement of Biomphalaria glabrata by the snail Thiara granifera in field habitats in St. Lucia, West Indies.

Thiara granifera is a melaniid snail capable of maintaining very high densities in a variety of habitats. It has been introduced into the New World from the Far East and is now spreading rapidly throughout the Caribbean. In Puerto Rico and Dominica casual observations following natural invasion by T. granifera suggest that it may exert a powerful restraining influence on populations of Biomphalaria glabrata, the major intermediate host of Schistosoma mansoni in the Caribbean. The potential of T. granifera in biological control is being investigated in St. Lucia. In four field trials, B. glabrata was apparently eliminated from marshes and streams six to 22 months after the introduction of T. granifera. Thiara granifera shows promise as a major factor in the suppression of schistosomiasis in the Caribbean, but it is unsuitable for universal use as it is an intermediate host of the lung fluke, Paragonimus westermani.

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Experimental maintenance of the schistosome vector Biomphalaria glabrata on an improved alginate gel.

An improved alginate formulation containing Spirulina algae or freeze-dried lettuce is described for experimental maintenance of Biomphalaria glabrata. Snail growth, food consumption and assimilation were higher on the gel containing Spirulina. The increased growth was not entirely the result of the higher food consumption observed with Spirulina, but appeared to be due to nutritional differences between the two nutrient sources. Snails responded to decreased dietary Spirulina concentration by increasing their relative level of assimilation rather than by increasing food consumption. An antibiotic treatment with chloramphenicol and ampicillin for the reduction of bacterial contaminants associated with B. glabrata is also described.

Alginates↗

The effect of schistosome infection on the mortality rates of Bulinus globosus and Biomphalaria pfeifferi.

Schistosoma infections of Bulinus globosus increased the per capita mortality rate in the laboratory from 0.231 to 0.406 per week. Schistosoma mansoni infections of Biomphalaria pfeifferi increased the per capita mortality rate in the laboratory from 0.055 to 0.177 per week. Calculations based on data from previous work by another author indicate an increase in the mortality rate of B. pfeifferi associated with prepatent S. mansoni infection.

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Interaction in the laboratory between Helisoma duryi, a possible competitor snail, and Biomphalaria pfeifferi, snail intermediate host of Schistosoma mansoni.

The planorbid snail, Helisoma duryi, has been proposed as a possible biocontrol agent against the intermediate host snails of schistosomiasis. In an attempt to evaluate the possible effect of interference between H. duryi and Biomphalaria pfeifferi, life-tables were compiled for these two snail species maintained both in the presence of each other and separately. The calculated population parameters indicated that H. duryi had a higher innate capacity of increase, net reproduction rate and finite rate of increase as well as a longer lifespan than the target species. However, the biological potential of B. pfeifferi was not influenced by the presence of H. duryi, and the latter appears to be unable to act as competitor against B. pfeifferi under similar prevailing conditions. It is possible that the situation might change under conditions of crowding and starvation.

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Predation and control of laboratory populations of the snail Biomphalaria glabrata by the freshwater prawn Macrobrachium rosenbergii.

Laboratory experiments were conducted on predation by the giant freshwater prawn, Macrobrachium rosenbergii, on Biomphalaria glabrata, a snail intermediate host of human schistosomiasis. Prawns greater than 22 mm carapace length could consume snails of any size. Smaller prawns exhibited a size-specific upper limit on the size of snail they could kill. Below the maximum size of prey consumed, little selectivity for prey size was demonstrated. All prawns heavier than 2.5 g killed the largest size of snail offered (16 mm diameter). Consumption, in terms of the numbers of snails killed, and the snails' wet weight and percentage of prawn body weight eaten per day, increased with prawn size. Large prawns consumed snails at a high rate (39% of body weight per day). Population experiments demonstrated that a prawn of 25 mm carapace length could eliminate 95% of a population of 80 snails in a 20-1 aquarium within 20 days and all snails by day 40. A prawn of 15 mm carapace length could not eliminate all snails. Large snails (24% of the initial population) had some protection from predation because of their size. Although the snails bred continuously, no snails were able to recruit to these populations in the presence of small prawns. In contrast, snail populations in control aquaria without prawns expanded to a mean of 919 snails by the end of the experiment (day 70). Since M. rosenbergii are voracious predators on B. glabrata, exhibit considerable habitat overlap with the snail prey, and are likely to treat snails as highly preferred food, further experimentation on these prawns in the context of biological control of schistosomiasis is warranted. The ready availability of prawns through established prawn hatcheries and the synergistic use of these prawns in aquaculture may ensure the reliability of stocking procedures and meet the standards of availability and cost-effectiveness required of a biological control agent. Polyculture of Macrobrachium with fin fish (Tilapia) indicates that these prawns may be a particularly useful control agent in aquacultural environments.

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Shade as a means of ecological control of Biomphalaria pfeifferi.

Species of Biomphalaria are attracted to light and negatively affected by darkness. Artificial shading of a breeding-site eliminated a population of B. pfeifferi within six weeks. The site was recolonized by eight weeks after the shade was removed. The time taken for recolonization suggests that shade acts not only by affecting snail behaviour but also indirectly by removing the diatoms which are food for the snails. The use of natural shade for ecological control of some snail vectors of schistosomiasis deserves, at least, careful consideration.

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The effect of exposure to selected numbers of Schistosoma mansoni miracidia on survival of cohorts of Biomphalaria pfeifferi.

Cohorts of 25 specimens of Biomphalaria pfeifferi were each exposed to batches of one, three, five, seven and 10 Schistosoma mansoni miracidia per snail. The effect on survival was evaluated by calculating the weekly per capita mortality rates and the mean life expectancy of the exposed snails. No statistically significant difference could be demonstrated between the mean per capita mortality rates of cohorts exposed to one and three miracidia per snail. Likewise there was no significant difference between the values obtained for cohorts exposed to five, seven and 10 miracidia per snail. In general, however, survival of the cohorts showed an inverse relationship with the number of miracidia to which each individual snail was exposed.

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Schistosoma mansoni: effect of aestivation on the intra-molluscan stages and the survival rate of infected Biomphalaria pfeifferi.

Following exposure to the miracidia of Schistosoma mansoni, Biomphalaria pfeifferi were induced to aestivate immediately ('0 h') or on days 7, 14 or 21 post-infection and then re-activated at various times. The proportions of snails found to be alive at weekly intervals after re-activation were found to be related to the length of time the snails had spent between infection and aestivation (I-A), varying from 100% (0 h) to 5% (21 days). The mean numbers of cercariae produced by the snails after re-activation also varied with I-A, being highest in those that aestivated immediately post-infection, and cercarial development was delayed by prolonged desiccation of the infected snails, the intra-molluscan development of S. mansoni is affected by the aestivation of the snail host.

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Extensive genetic variation revealed in adjacent populations of the schistosome intermediate host Biomphalaria pfeifferi from a single river system.

Random amplified polymorphic DNA (RAPD) analysis was used to study genetic variation among schistosome hosts of the species Biomphalaria pfeifferi. The analysed snails came from 15 sites along a 6-km stretch of a Zimbabwean river, providing data on genetic-geographical relationships over, probably, the smallest scale yet studied for any snail intermediate host species. Only 6% of the DNA fragments were common to all snails, and snail populations as geographically close as 100 m were genetically distinct. The most genetically polymorphic snail populations were those collected within and downstream from human water-contact sites.

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Experimental control of the schistosome-transmitting snail Biomphalaria pfeifferi by the ampullariid snail Pila ovata.

Adults of the African ampullariid snail Pila ovata were examined for their ability to control laboratory populations of the pulmonate snail Biomphalaria pfeifferi, a widespread, intermediate host of the human pathogen Schistosoma mansoni in sub-Saharan Africa. In a 6-week experiment conducted in large (100 x 60 x 60 cm) outdoor tanks containing floating macrophytes (Nymphaea caerula) and initially set up with one adult ampullariid for every three adult pulmonates, the numbers of B. pfeifferi egg masses were always about half those in similar tanks without P. ovata. Although, by week 6, the numbers of B. pfeifferi in the control tanks (without ampullariids) had increased 5-fold, from an initial mean of 30 snails/tank, there was no significant increase in the numbers of B. pfeifferi in the experimental tanks (containing ampullariids). Results of experiments conducted in indoor glass aquaria indicated that adult P. ovata rapidly attacked egg masses or neonates (< 2.5 mm shell diameter) of B. pfeifferi but had no effect on the adults. The adult ampullariids also significantly decreased cover by floating macrophytes over a 6-week period compared with that in similar but ampullariid-free aquaria. This decrease in plant cover is relevant to biological control of the schistosome vectors as macrophytes serve as food, shelter and oviposition sites for pulmonate snails. The present result indicate the ability of P. ovata to inhibit multiplication of B. pfeifferi populations, at least under laboratory conditions, both directly, through predation, and indirectly, by competition for resources. Pila ovata may therefore prove useful in the biological control of medically important, pulmonate snails.

Animals↗

Effect of aestivation of Biomphalaria pfeifferi on the survival and infectivity of Schistosoma mansoni cercariae.

Schistosoma mansoni cercariae from post-aestivated Biomphalaria pfeifferi remain motile for 20 hours after release. Thereafter, their activity decreases with age. The difference in mortality rate of cercariae from aestivated and non-aestivated B. pfeifferi studied here proved to be statistically significant (P < 0.05) within the first 10 hours of the experimental period. Results of the percentage recovery of worms from different mouse organs infected with cercariae from aestivated and non-aestivated snails varied. The two main organs infected were the liver and intestine. In conclusion, the penetration, migration and maturation of cercariae into adult worms were not affected by the aestivation of B. pfeifferi.

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Structural characterization of N-glycans from the freshwater snail Biomphalaria glabrata cross-reacting with Schistosoma mansoni glycoconjugates.

The human parasitic trematode Schistosoma mansoni has a complex life cycle that includes the freshwater snail Biomphalaria glabrata as intermediate host. Within each stage, the parasite synthesizes a wide array of glycoconjugates, exhibiting, in part, unique carbohydrate structures. In addition, the parasite expresses definitive host-like sugar epitopes, such as Lewis X determinants, supporting the concept of carbohydrate-mediated molecular mimicry as an invasion and survival strategy. In the present study, we investigated whether common carbohydrate determinants occur also at the level of the intermediate host. To this end, a structural characterization of hemolymph glycoprotein-N-glycans of B. glabrata was performed. N-glycans were released from tryptic glycopeptides and labeled with 2-aminopyridine. Sugar chains serologically cross-reacting with S. mansoni glycoconjugates were isolated by immunoaffinity chromatography using a polyclonal antiserum directed against schistosomal egg antigens and fractionated by Aleuria aurantia lectin affinity chromatography and high-performance liquid chromatography. Obtained glycans were analyzed by different mass spectrometric techniques as well as by monosaccharide constituent and linkage analysis. The results revealed a highly heterogeneous oligosaccharide pattern. Cross-reacting species represented about 5% of the total glycans and exhibited a terminal Fuc(alpha1-3)GalNAc unit, a (1-2)-linked xylosyl residue, or both types of structural motifs. In conclusion, our study demonstrates the presence of common carbohydrate epitopes also at the level of S. mansoni and its intermediate host.

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Immune-recognition of Schistosoma mansoni primary sporocysts may require specific receptors on Biomphalaria glabrata hemocytes.

Cellular interactions, leading to cell-mediated cytotoxicity when Biomphalaria glabrata hemocytes encapsulate Schistosoma mansoni sporocysts, have been investigated. Rabbit antibodies (IgG), when bound to antigens on sporocyst surfaces, prevent the normal cytoadherence (CA) of hemocytes from both susceptible and resistant host snails. Since interference with CA occurs with even fixed sporocysts, the effect is not due to IgG stimulated modulation of the parasite surface. Using two antisera with some overlapping specificities, and quantitative immunofluorescent antibody technique (QIFAT), we determined the concentrations of IgG needed to place equivalent amounts of IgG on the sporocysts. At these concentrations, CA was affected differentially, implying that interference was due to the specific antigens bound, and not due simply to the presence of IgG. Also with QIFAT we determined how much F(ab')2 and IgG from anti-sporocyst serum were needed to block an equivalent amount of antigenic determinants from access by whole FITC labelled IgG. Sporocysts whose surface antigens were equally blocked were equally unadherent for hemocytes, supporting the notion that the nature of obscured antigens, and neither the Fc portion nor the larger size of intact IgG protein, was responsible for the effect on CA. These surprising results imply a role for specific antigen binding sites on snail hemocytes.

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Opsonization of yeast by the plasma of Biomphalaria glabrata (Gastropoda): a strain-specific, time-dependent process.

Phagocytosis of yeast by haemocytes of Biomphalaria glabrata, intermediate host of Schistosoma mansoni, is influenced strongly by host plasma components, although it can occur without involvement of such factors. Plasma from two strains of B. glabrata which are resistant to S. mansoni differs in its opsonic properties from the plasma of a susceptible strain. This may reflect the principles which determine compatibility or incompatibility in this host-parasite system. Opsonization is a time-dependent process: short periods of incubation in plasma from all strains reduces subsequent phagocytosis in the absence of plasma factors, whereas longer incubation in resistant strain plasma is markedly opsonic. Haemocytes from all strains examined are equally competent in their recognition of either native or opsonized yeast.

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Resistance to Schistosoma mansoni by transplantation of APO Biomphalaria tenagophila.

Transplantation of the haematopoietic organ from Biomphalaria tenagophila (Taim strain, RS, Brazil), resistant to Schistosoma mansoni, to a highly susceptible strain (Cabo Frio, RJ, Brazil) of the same species, showed in the recipient snails resistance against the trematode, when a successful transplant occurred. The success of transplantation could be confirmed by a typical molecular marker of the Taim strain in haemocytes of the recipients (350 bp detected by PCR-RFLP). The recipient snails which did not present the donor marker in haemocytes (unsuccessful transplantation) were infected with the parasite. The use of an atoxic modelling clay for closing the hole in the transplantation site reduced significantly the mortality caused by bleeding after transplantation procedures.

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Schistosoma mansoni: identification of chemicals that attract or trap its snail vector, Biomphalaria glabrata.

A new bioassay for chemical attractants of aquatic snails demonstrated that Biomphalaria glabrata could be attracted to or trapped in the vicinity of homogenates of lettuce. Fractionation of homogenates revealed the amino acids glutamate and proline and the primary attractants. Attraction was specific for the L form of glutamate. Proline appeared to stimulate reproductive activity. Glutathione, gamma-aminobutyric acid, and a number of other compounds had no effect. Extracts of lyophilized snail tissue also attracted other snails and may thus contain pheromones. These results permit formulation and testing of controlled-release attractants designed to overcome the repellant effects of slow-release molluscicides, as well as the design of stimulants to be used with no-release poisons.

Amino Acids↗