Competitive interactions between species of fresh-water snails. I. Laboratory. IC. Comparative survival of Biomphalaria glabrata and B. straminea kept out of water

Biomphalaria glabrata and B. straminea were submitted to an out-door laboratory experiment for testing their comparative ability to resist desiccation. Results have shown that B. straminea is significantly higher resistant than B. glabrata. After five months under such distressing condition the survival ratios were: B. glabrata 8.1 per cent and B. straminea 18.4 per cent.     

Intraspecific variations in the hemolymph of Biomphalaria glabrata, a snail host of Schistosoma mansoni.

An attempt was made to characterize the hemolymph of Biomphalaria glabrata with reference to "normal" intra-specific variation, i.e., both inter- and intra-strain differences. Total protein concentration, per cent hemoglobin, pH, and osmolarity were studied. Seven geographic strains of B, glabrata were examined. In addition, observations were made on the hemolymph of Biomphalaria straminea, several strains of Helisoma caribaeum, and on B. glabrata subjected to infection with Schistosoma mansoni or to periods of starvation. Intra-strain differences in total protein concentration and total hemoglobin concentration in B. glabrata appeared to be more closely related with snail size than with absolute age. Inter-strain variation in B. glabrata was also noted, but the differences were of the same magnitude as those from intra-strain samples. Significant differences in total protein concentration were observed, however, between the means of similar size B. glabrata, B. straminea and H. caribaeum. The osmolatity of the hemolymph from different size B. glabrata was similar as were the osmolalities of the hemolymph from similar size snails of different strains. However, all B. glabrata strains exhibited hemolymph osmolalities lower than observed in strains of H. caribaeum. Infection with S. mansoni reduced the protein concentration of B. glabrata hemolymph. Differences were noted as early as 1.5-24 hr post-infection, with significant alterations occurring at about 11 days post-infection. To a lesser extent, starvation also depleted the protein content of the hemolymph.     

Geographical distribution of Biomphalaria snails in the State of Minas Gerais, Brazil

Published and unpublished observations on geographical distribution of Biomphalaria snails in the State of Minas Gerais, Brazil, were compiled. This work is aimed at knowing the present occurrence of Biomphalaria species in this region, and at contributing to the elaboration of the planorbid chart of Minas Gerais. In malacological surveys, performed by several researchers, the presence of seven species of this genus was recorded. Those planorbids were found in 12 mesoregions, in 283 (33.1%) municipalities out of 853 with the following distribution: B. glabrata (185 municipalities), B. straminea (125), B. tenagophila (58), B. peregrina (57), B. schrammi (26), B. intermedia (20) and B. occidentalis (2). B. glabrata and B. tenagophila are found naturally infected by Schistosoma mansoni in Minas Gerais. In 24 municipalities the three snail hosts of S. mansoni in Brazil, B. glabrata, B. tenagophila and B. straminea, are present.     

Dispersion of Biomphalaria straminea in the State of Sergipe: a comparative study within an interval of 19 years

The aim of the current paper was to study in the state of Sergipe the population distribution and interactions between two species of Biomphalaria, the snail vectors of Schistosoma mansoni in the northeastern Brazil. Data collected in 1969 showed that B. straminea, with only one exception, was limited to the semi-dry region, while B. glabrata was found to live exclusively in the forest region, both in the state of Sergipe. This spatial distribution seemed to suggest that the above Biomphalaria species used to dominate specific territories. Snail collections made in 1988 in the same 37 places searched in 1969, showed that B. straminea has invaded territories previously occupied by B. glabrata, suggesting that a process of competitive displacement is taking place between these two closely related species. Natural snail infection rates were determined and some ecological aspects of the snail breeding places were registered.     

Longitudinal Study on the Natural Infection of Biomphalaria straminea and B. glabrata by Schistosoma mansoni in an Endemic Area of Schistosomiasis in Pernambuco,Brazil

The abundance of snail hosts and the rates of infection with Schistosoma mansoni were monitored monthly for four years in two representative localities subjected to repeated chemotherapy of infected persons. Snail abundance varied from 1.0 to 4.4 collected per person/minute/station for Biomphalaria straminea and from 0.1 to 7.0 for B. glabrata. Infection rates of snails in nature varied from 0% to 15% for the former and from 0% to 70% for the latter species. Human infection increased from 35.5% to 61.9% in the locality occupied by B. straminea, and decreased from 40.3% to 20.8% in that occupied by B. glabrata. No relationship could be detected between human infection and the snail variables. Despite seasonal variations, natural infection persisted throughout the monitoring period in both snail species. It reached remarkably high levels in B. straminea when compared to those obtained by other authors probably because of differences in methodology. It is recommended that longitudinal studies should be carried out focally and periodically to avoid underestimating the prevalence of schistosome infection in snails.     

Genetic variability in Brazilian populations of Biomphalaria straminea complex detected by simple sequence repeat anchored polymerase chain reaction amplification

Biomphalaria glabrata, B. tenagophila and B. straminea are intermediate hosts of Schistosoma mansoni, in Brazil. The latter is of epidemiological importance in the northwest of Brazil and, due to morphological similarities, has been grouped with B. intermedia and B. kuhniana in a complex named B. straminea. In the current work, we have standardized the simple sequence repeat anchored polymerase chain reaction (SSR-PCR) technique, using the primers (CA)8RY and K7, to study the genetic variability of these species. The similarity level was calculated using the Dice coefficient and genetic distance using the Nei and Li coefficient. The trees were obtained by the UPGMA and neighbor-joining methods. We have observed that the most related individuals belong to the same species and locality and that individuals from different localities, but of the same species, present clear heterogeneity. The trees generated using both methods showed similar topologies. The SSR-PCR technique was shown to be very efficient in intrapopulational and intraspecific studies of the B. straminea complex snails.     

Polymerase chain reaction and restriction fragment length polymorphism of cytocrome oxidase subunit I used for differentiation of Brazilian Biomphalaria species intermediate host of Schistosoma mansoni

The intermediate hosts of Schistosoma mansoni, in Brazil, Biomphalaria glabrata, B. tenagophila and B. straminea, were identified by restriction fragment length polymorphism analysis of the mitochondrial gene cytochrome oxidase I (COI). We performed digestions with two enzymes (AluI and RsaI), previously selected, based on sequences available in Genbank. The profiles obtained with RsaI showed to be the most informative once they were polymorphic patterns, corroborating with much morphological data. In addition, we performed COI digestion of B. straminea snails from Uruguay and Argentina.     

First report on the presence of Biomphalaria straminea in the municipality of Jaboticatubas,State of Minas Gerais,Brazil

This is the first report on occurrence of Biomphalaria straminea in the district of S?o José de Almeida (municipality of Jaboticatubas) State of Minas Gerais, Brazil. The presence of B. glabrata and B. tenagophila had already been reported in this area. Such municipality is part of the metropolitan region of Belo Horizonte and comprises 60% of the Tourist Complex of Serra do Cipó. Since the 1950s throughout the 1990s, a schistosomiasis prevalence ranging from 15 to 40% has been observed. Although no B. straminea specimen has been found naturally infected in the region, descendants of these snails collected in the area, showed to be experimentally susceptible to Schistosoma mansoni infection reaching rates from 14.6 to 28.6%. Even not being found naturally infected, in the State of Minas Gerais, the possibility that the species B. straminea may keep endemicity foci of schistosomiasis should be regarded, as in the Northeastern region of Brazil where the high density of this planorbid and the social-economic and sanitary conditions enable to the transmission.     

Morphological and polymerase chain reaction-restriction fragment lenght polymorphism characterization of Biomphalaria kuhniana and Biomphalaria amazonica from Colombia

In Colombia, five Biomphalaria planorbid species are known: B. kuhniana, B. straminea, B. peregrina, B. canonica and B. oligoza(var. B. philippiana). Among them, B. straminea is intermediate host of Schistosoma mansoni and B. peregrina has been found to be experimentally susceptible to this parasite. B. straminea is commonly confused with B. kuhniana and they have been clustered together with B. intermedia in the complex named B. straminea. The difficulties involved in the specific identification, based on morphological data, have motivated the use of new techniques as auxiliary tools in cases of inconclusive morphological identification of such planorbid. In the present study, five Biomphalaria populations from the Colombian Amazon region and from Interandian Valleys were morphologically identified and characterized by polymerase chain reaction-restriction fragment lenght polymorphism directed at the internal transcribed spacer region of the rRNA gene, followed by digestion of the generated fragment with restriction enzymes (DdeI, AluI, RsaI, MvaI and HaeIII). Known profiles of the Brazilian species B. straminea, B. peregrina, B. kuhniana, B. intermedia and B. amazonica, besides B. kuhniana from Colombia, were used for comparison. The five populations under study were morphologically and molecularly identified as B. kuhniana and B. amazonica.     

Orientation of the snail Biomphalaria straminea (Dunker, 1848) in response to light in a situation of selection

The behavioral response of Biomphalaria straminea to light was evaluated in terms of location of the snail in a Y-shaped aquarium in a situation of selection and of the rate (cm/hour) and direction of locomotion under homogeneous (vertical) or differential (horizontal) lighting upon only one arm of the aquarium. The light source consisted of daylight fluorescent lamps with a spectrum close to that of natural light, with illumination varying from 2.8 to 350 lux. Analysis of the data showed that all animals, whether in groups or isolated, were attracted to light, although the time needed to approach the light source was 50% shorter for the former than for the latter. The rate of locomotion of B. straminea was 35% higher than that observed in B. glabrata and 51% higher than that observed in B. tenagophila studied under similar conditions. The results are discussed in terms of social factors and geographical distribution of the three species.     

Schistosoma mansoni: use of a cloned ribosomal RNA gene probe to detect restriction fragment length polymorphisms in the intermediate host Biomphalaria glabrata.

Adult susceptibility of Biomphalaria glabrata to Schistosoma mansoni infection is controlled by simple Mendelian genetics. In this study a molecular approach was used to determine the degree of genetic variation between well-defined lines of B. glabrata which are either resistant (10-R2) or susceptible (M-line) to S. mansoni infection. A cloned probe pSM389, which contains part of the S. mansoni small ribosomal RNA gene and a portion of the nontranscribed spacer was found to cross-hybridize with B. glabrata DNA and was used in Southern hybridizations to detect restriction fragment length polymorphisms (RFLPs) between the above snail stocks. Polymorphisms were noted with a variety of restriction enzymes, namely Bg/II, BamHI, AccI, AvaII, ClaI, EcoRI, EcoRV, KpnI, PvuII, and NcoI. Although most RFLPs were relatively minor, a significant difference was observed with EcoRV. Further analysis of the EcoRV RFLPs among other isolates of the resistant stock demonstrated that a high frequency of genetic variation exists even among isolates of the same origin, but maintained in separate laboratories. Interestingly, RFLPs in the EcoRV site were detected in DNA isolated from a single generation of selfed progeny of a single 10-R2 parent. RFLPs associated with this site were found to occur between B. glabrata and B. tenagophila, B. straminea, and B. schrammi, indicating that Southern blot analysis using ribosomal gene probes may be useful for the molecular differentiation of B. glabrata from other intermediate hosts and from morphologically similar species that are refractory to infection.     

Manganese superoxide dismutase from Biomphalaria glabrata

The investigation of the response of Biomphalaria glabrata snails to Echinostoma paraensei (digenea) at 2 days post-exposure by suppression subtractive hybridization yielded a partial sequence of the anti-oxidant enzyme manganese superoxide dismutase (MnSOD). Full-length MnSOD (669nt) from M line and BS-90 strains of B. glabrata differed by one synonymous nucleotide replacement. B. glabrata has 1-4 MnSOD loci (Southern hybridization). Both snail strains expressed MnSOD at equal baseline levels (quantitative PCR). Susceptible snails increased expression of MnSOD following infection with E. paraensei or Schistosoma mansoni, and expression was reduced in the incompatible combination (BS-90 B. glabrata and S. mansoni). Thus, MnSOD did not determine resistance or susceptibility for these parasites, but expression of MnSOD is consistent with its involvement in a stress response of B. glabrata.     

Evolutionary relationships and biogeography of Biomphalaria (Gastropoda: Planorbidae) with implications regarding its role as host of the human bloodfluke, Schistosoma mansoni.

The wide geographic distribution of Schistosoma mansoni, a digenetic trematode and parasite of humans, is determined by the occurrence of its intermediate hosts, freshwater snails of the genus Biomphalaria (Preston 1910). We present phylogenetic analyses of 23 species of Biomphalaria, 16 Neotropical and seven African, including the most important schistosome hosts, using partial mitochondrial ribosomal 16S and complete nuclear ribosomal ITS1 and ITS2 nucleotide sequences. A dramatically better resolution was obtained by combining the data sets as opposed to analyzing each separately, indicating that there is additive congruent signal in each data set. Neotropical species are basal, and all African species are derived, suggesting an American origin for the genus. We confirm that a proto-Biomphalaria glabrata gave rise to all African species through a trans-Atlantic colonization of Africa. In addition, genetic distances among African species are smaller compared with those among Neotropical species, indicating a more recent origin. There are two species-rich clades, one African with B. glabrata as its base, and the other Neotropical. Within the African clade, a wide-ranging tropical savannah species, B. pfeifferi, and a Nilotic species complex, have both colonized Rift Valley lakes and produced endemic lacustrine forms. Within the Neotropical clade, two newly acquired natural hosts for S. mansoni (B. straminea and B. tenagophila) are not the closest relatives of each other, suggesting two separate acquisition events. Basal to these two species-rich clades are several Neotropical lineages with large genetic distances between them, indicating multiple lineages within the genus. Interesting patterns occur regarding schistosome susceptibility: (1) the most susceptible hosts belong to a single clade, comprising B. glabrata and the African species, (2) several susceptible Neotropical species are sister groups to apparently refractory species, and (3) some basal lineages are susceptible. These patterns suggest the existence of both inherent susceptibility and resistance, but also underscore the ability of S. mansoni to adapt to and acquire previously unsusceptible species as hosts. Biomphalaria schrammi appears to be distantly related to other Biomphalaria as well as to Helisoma, and may represent a separate or intermediate lineage.     

Biological invasions: the case of planorbid snails

A large number of planorbid snails are now commonly transported by man mainly through the aquatic plant trade. However, only a restricted number of species establish viable populations in a new habitat and a more restricted number spread. Only five planorbid species can be ranked in this last category and can be considered as pests because of their role in the transmission of parasites to humans or domestic animals: Biomphalaria glabrata, B. straminea, B. tenagophila, B. pfeifferi and Indoplanorbis exustus. The neotropical B. glabrata, B. straminea and B. tenagophila have proven their capacity to invade another continent sometimes creating new transmission foci. The African B. pfeifferi and the Indian I. exustus have also expanded their distribution area with long-distance dispersal. Other planorbid species, i.e. Helisoma duryi, Amerianna carinata and Gyraulus spp. have been able to establish viable populations, but not to spread, presumably because they are limited to specific habitats or/and display poor competitive abilities.     

Displacement of Biomphalaria glabrata by Thiara granifera under natural conditions in the Dominican Republic.

After a study of the population dynamics of Biomphalaria glabrata snails in several breeding places in the Dominican Republic, the snail Thiara granifera was introduced in some B. glabrata habitats. T. granifera became established in one point in one habitat in the town of Quisqueya, in the east of the country. Around this point of establishment 6 points were selected in order to observe the population dynamics of both species of snails and the chemical and biological characteristics at each point. Four of these points already harbored B. glabrata. One control point was selected also harboring B. glabrata. After 14 months of observations, the results showed that T. granifera was competing with and displacing B. glabrata. This competition does not seem to be competition for food or vital space. Rather, B. glabrata avoids the presence of T. granifera and moves away to new areas, and this is possibly due to a chemical substance(s) secreted by T. granifera or by physical contact with the large number of individuals of T. granifera.     

Diagnostic of Biomphalaria snails and Schistosoma mansoni: DNA obtained from traces of shell organic materials

Freshwater snails belonging to the genus Biomphalaria act as intermediate hosts for the parasite trematode Schistosoma mansoni in Africa and in the neotropical region. Identification of such molluscs is carried out based on morphological characters and the presence of cercariae is verified through squeezing snails between two glass slides or by exposing them to artificial light. However, sometimes, the material collected includes molluscs with decomposed bodies or, yet, only empty shells, which precludes their identification and S. mansoni detection. Due to these difficulties, we have developed a methodology in which DNA may be extracted from traces of organic material from inside shells in order to identify molluscs through polymerase chain reaction and restriction fragment length polymorphism and to detect S. mansoni into these snails, by using low stringency polymerase chain reaction. Species-specific profiles obtained from B. glabrata, B. straminea, and B. tenagophila snails and their shells, maintained in laboratory for ten years, showed the same profiles. S. mansoni profiles showed to be present in shell specimens as far as the eighth week after being removed from aquarium.     

Molecular studies of Biomphalaria glabrata, an intermediate host of Schistosoma mansoni

The freshwater gastropod Biomphalaria glabrata is one of the most important invertebrate hosts of the helminth parasite Schistosoma mansoni. Investigators are using different strategies to determine the molecular basis of this snail-parasite relationship. Of particular interest are the identification of parasite resistance genes in the snail, and the application of molecular probes to better understand the epidemiology of schistosomiasis. This review will focus on recent advances that have been made on genome analysis of B. glabrata. Much of this work has centred on the use of random amplification of polymorphic DNA-PCR-based technology, with restriction fragment length polymorphism analysis and the generation of expressed sequence tags from the snail. A brief discussion of how parasite products may complicate this analysis is also given, along with an indication of the scope of the problems that lie ahead.     

Malacological assessment and natural infestation of Biomphalaria straminea (Dunker, 1848) by Schistosoma mansoni (Sambon, 1907) And Chaetogaster limnaei (K. Von Baer, 1827) in an urban eutrophic watershed.

The objective of this study was to perform a malacological assessment at the Ibirité reservoir watershed in the metropolitan region of Belo Horizonte (Minas Gerais) and to evaluate the natural infestation rate of Biomphalaria straminea (Gastropoda: Planorbidae) by Schistosoma mansoni (Platyhelminthes: Trematoda) and Chaetogaster limnaei (Oligochaeta: Naididae). The samples were collected from July to August 2002. The B. straminea individuals collected were kept in the laboratory; the natural infestation rate by S. mansoni and C. limnaei was assessed weekly. The malacological assessment identified five mollusk species present in the Ibirité reservoir watershed: B. straminea, Physa marmorata, Lymnea sp., Melanoides tuberculatus, and Pomacea austrum. Laboratory observations showed that the B. straminea individuals were infected by C. limnaei rather than S. mansoni. Although there was no infection of B. straminea by S. mansoni, presence of B. straminea in itself merits close attention due to possible risk of human schistosomiasis by the local population.     

Circadian Rhythms in the Cercarial Emergence of Schistosoma mansoni by Biomphalaria tenagophila at Outdoors: A Comparative Study with Biomphalaria glabrata

Circadian rhythms in the emergence of S. mansoni cercariae from Biomphalaria tenagophila and B. glabrata, snail hosts of schistosomosis in Brazil, were investigated. A total of 35 specimens of B. tenagophila (São Paulo, Brazil) and 12 B. glabrata (Minas Gerais, Brazil) exposed individually to five miracidia of Schistosoma mansoni originated from the same biotope as their snail hosts, were tested. Observations were carried out at outdoors, with the quantification of cercarial emergence at 3h intervals during three consecutive days in November 1989 and in May 1990. Cercarial emergence was essentially diurnal (from 06.00-18.00h) in both species. Circadian rhythms were detected by the Single Cosinor Method among 74.3% of B. tenagophila and 91.7% of B. glabrata snails. The acrophases corresponding to individual snails were between 11.37 e 17.54h in B. tenagophila and between 14.15 and 16.29h in B. glabrata. These findings confirm our preliminary observations in B. tenagophila and are in accordance to those of other authors in regard to B. glabrata. The acrophases of individual snails were similar within each species, thus indicating that at populacional level cercarial emergence was concentrated in particular times of the day. Group acrophases for each species varied from 13.22 to 15.22h and were not significantly different between B. tenagophila and B. glabrata. Cercariae emerging from B. tenagophila snails seemed to be more sensitive to environmental temperature than those emerging from B. glabrata, at least in the temperature range prevailing along the tests. Further chronobiological studies on host-parasite interactions are encouraged to improve our knowledge about temporal aspects of schistosomosis transmission.     

Differential lectin labelling of circulating hemocytes from Biomphalaria glabrata and Biomphalaria tenagophila resistant or susceptible to Schistosoma mansoni infection

Lectins/carbohydrate binding can be involved in the Schistosoma mansoni recognition and activation of the Biomphalaria hemocytes. Therefore, expression of lectin ligands on Biomphalaria hemocytes would be associated with snail resistance against S. mansoni infection. To test this hypothesis, circulating hemocytes were isolated from B. glabrata BH (snail strain highy susceptible to S. mansoni), B. tenagophila Cabo Frio (moderate susceptibility), and B. tenagophila Taim (completely resistant strains), labelled with FITC conjugated lectins (ConA, PNA, SBA, and WGA) and analyzed under fluorescence microscopy. The results demonstrated that although lectin-labelled hemocytes were detected in hemolymph of all snail species tested, circulating hemocytes from both strains of B. tenagophila showed a larger number of lectin-labelled cells than B. glabrata. Moreover, most of circulating hemocytes of B. tenagophila were intensively labelled by lectins PNA-FITC and WGA-FITC, while in B. glabrata small hemocytes were labeled mainly by ConA. Upon S. mansoni infection, lectin-labelled hemocytes almost disappeared from the hemolymph of Taim and accumulated in B. glabrata BH. The role of lectins/carbohydrate binding in resistance of B. tengophila infection to S. mansoni is still not fully understood, but the data suggest that there may be a correlation to its presence with susceptibility or resistance to the parasite.