Species diversity and distribution of schistosome intermediate snail hosts in The Gambia.

There is a need for recent information on intermediate snail hosts of schistosomes in The Gambia; the previous studies were conducted over three decades ago. This study assessed the incidence, species diversity, distribution and infection status of schistosome intermediate snail hosts in the country. Malacological surveys were conducted in all 5 regions of The Gambia: Central River Region (CRR), Upper River Region (URR), Western Region (WR), Lower River Region (LRR) and North Bank Region (NBR). Sampling of snails was undertaken at 114 sites that included permanent water bodies such as streams (bolongs), rice fields, irrigation canals and swamps; and temporal (seasonal) laterite pools. Ecological and physicochemical factors of sites were recorded. Snails were identified morphologically and screened for schistosome infections using molecular techniques. Freshwater snails were found at more than 50% (60/114) of sites sampled. While three species of Bulinus were collected, no Biomphalaria snails were found in any of the sites sampled. Of the total 2877 Bulinus snails collected, 75.9% were identified as Bulinus senegalensis, 20.9% as Bulinus forskalii and 3.2% as Bulinus truncatus. Seasonal pools produced the largest number of snails, and CRR was the region with the largest number of snails. Bulinus senegalensis was found more in seasonal pools as opposed to permanent sites, where B. forskalii and B. truncatus were observed to thrive. Bulinus snails were more common in seasonal sites where aquatic vegetation was present. In permanent sites, the abundance of snails increased with increase in water temperature and decrease in water pH. Bulinus senegalensis was found infected with both S. haematobium and S. bovis, while B. forskalii and B. truncatus had only S. bovis infection. While the human parasite S. haematobium was restricted to just four sites, the livestock parasite S. bovis had a much more widespread geographical distribution across both CRR and URR. This new information on the distribution of intermediate snail hosts of schistosomes in The Gambia will be vital for the national schistosomiasis control initiative.

Assessing the diversity and distribution of potential intermediate hosts snails for urogenital schistosomiasis: Bulinus spp. (Gastropoda: Planorbidae) of Lake Victoria.

BACKGROUND: The Lake Victoria basin is one of the most persistent hotspots of schistosomiasis in Africa, the intestinal form of the disease being studied more often than the urogenital form. Most schistosomiasis studies have been directed to Schistosoma mansoni and their corresponding intermediate snail hosts of the genus Biomphalaria, while neglecting S. haematobium and their intermediate snail hosts of the genus Bulinus. In the present study, we used DNA sequences from part of the cytochrome c oxidase subunit 1 (cox1) gene and the internal transcribed spacer 2 (ITS2) region to investigate Bulinus populations obtained from a longitudinal survey in Lake Victoria and neighbouring systems during 2010-2019. METHODS: Sequences were obtained to (i) determine specimen identities, diversity and phylogenetic positions, (ii) reconstruct phylogeographical affinities, and (iii) determine the population structure to discuss the results and their implications for the transmission and epidemiology of urogenital schistosomiasis in Lake Victoria. RESULTS: Phylogenies, species delimitation methods (SDMs) and statistical parsimony networks revealed the presence of two main groups of Bulinus species occurring in Lake Victoria; B. truncatus/B. tropicus complex with three species (B. truncatus, B. tropicus and Bulinus sp. 1), dominating the lake proper, and a B. africanus group, prevalent in banks and marshes. Although a total of 47 cox1 haplotypes, were detected within and outside Lake Victoria, there was limited haplotype sharing (only Haplotype 6 was shared between populations from Lake Victoria open waters and neighbouring aquatic systems) - an indication that haplotypes are specific to habitats. CONCLUSIONS: The Bulinus fauna of Lake Victoria consists of at least B. truncatus, B. tropicus, Bulinus sp. 1 (B. trigonus?) and B. ugandae. The occurrence and wide distribution of Bulinus species in Lake Victoria potentially implies the occurrence of urogenital schistosomiasis in communities living along the shores and on islands of the lake who depend solely on the lake for their livelihood. More in-depth studies are needed to obtain a better picture of the extent of the disease in the Lake Victoria basin.

Molecular identification of Bulinus spp. intermediate host snails of Schistosoma spp. in crater lakes of western Uganda with implications for the transmission of the Schistosoma haematobium group parasites.

BACKGROUND: Human schistosomiasis is the second most important tropical disease and occurs in two forms in Africa (intestinal and urogenital) caused by the digenetic trematodes Schistosoma mansoni and Schistosoma haematobium, respectively. A proposed recent shift of schistosomiasis above a previously established altitudinal threshold of 1400 m above sea level in western Ugandan crater lakes has triggered more research interest there. METHODS: Based on extensive field sampling in western Uganda and beyond and employing an approach using sequences of the mitochondrial barcoding gene cytochrome c oxidase subunit 1 (cox1) this study aims were: (i) identification and establishment of the phylogenetic affinities of Bulinus species as potential hosts for Schistosoma spp.; (ii) determining diversity, frequency and distribution patterns of Bulinus spp.; and (iii) establishing genetic variability and phylogeographical patterns using Bayesian inference and parsimony network analyses. RESULTS: Out of the 58 crater lakes surveyed, three species of Bulinus snails were found in 34 crater lakes. Bulinus tropicus was dominating, Bulinus forskalii was found in two lakes and Bulinus truncatus in one. The latter two species are unconfirmed potential hosts for S. haematobium in this region. However, Bulinus tropicus is an important species for schistosomiasis transmission in ruminants. Bulinus tropicus comprised 31 haplotypes while both B. forskalii and B. truncatus exhibited only a single haplotype in the crater lakes. All species clustered with most of the haplotypes from surrounding lake systems forming source regions for the colonization of the crater lakes. CONCLUSIONS: This first detailed malacological study of the crater lakes systems in western Uganda revealed presence of Bulinus species that are either not known or not regionally known to be hosts for S. haematobium, the causing agent of human urogenital schistosomiasis. Though this disease risk is almost negligible, the observed dominance of B. tropicus in the crater lakes shows that there is a likelihood of a high risk of infections with Schistosoma bovis. Thus, extra attention should be accorded to safeguard wild and domestic ruminants in this region as the population benefits from these animals.

Mapping freshwater snails in north-western Angola: distribution, identity and molecular diversity of medically important taxa.

BACKGROUND: This study was designed to determine the distribution and identity of potential intermediate snail hosts of Schistosoma spp. in Bengo, Luanda, Kwanza Norte and Malanje Provinces in north-western Angola. This is an area where infection with Schistosoma haematobium, causing urogenital schistosomiasis, is common but little is yet known about transmission of the disease. Angola has had a varied past with regard to disease control and is revitalising efforts to combat neglected tropical diseases. METHODS: Snails were sampled from 60 water-contact points. Specimens of the genera Bulinus, Biomphalaria or Lymnaea were screened for trematode infections by inducing cercarial shedding. Snails were initially identified using shell morphology; subsequently a cytochrome c oxidase subunit 1 (cox1) gene fragment was amplified from a subset of snails from each site, for molecular identification. Cercariae were captured onto FTA cards for molecular analysis. Specimens of Bulinus angolensis collected from the original locality of the type specimen have been characterised and comparisons made with snails collected in 1957 held at the Natural History Museum, London, UK. RESULTS: In total snails of nine genera were identified using morphological characteristics: Biomphalaria, Bulinus, Gyraulus, Lanistes, Lentorbis, Lymnaea, Melanoides, Physa and Succinea. Significant for schistosomiasis transmission, was the discovery of Bulinus globosus, B. canescens, B. angolensis, B. crystallinus and Biomphalaria salinarum in their type-localities and elsewhere. Bulinus globosus and B. angolensis occurred in two distinct geographical areas. The cox1 sequence for B. globosus differed markedly from those from specimens of this species collected from other countries. Bulinus angolensis is more closely related to B. globosus than originally documented and should be included in the B. africanus group. Schistosoma haematobium cercariae were recovered from B. globosus from two locations: Cabungo, Bengo (20 snails) and Calandula, Malanje (5 snails). Schistosoma haematobium cercariae were identified as group 1 cox1 corresponding to the type common throughout the African mainland. CONCLUSIONS: Various freshwater bodies in north-western Angola harbour potential intermediate snail hosts for urogenital schistosomiasis, highlighting the need to map the rest of the country to identify areas where transmission can occur and where control efforts should be targeted. The molecular phylogeny generated from the samples confirmed that considerable variation exists in B. globosus, which is the primary snail host for S. haematobium in many regions of Africa.

Population genetic structure of the freshwater snail, Bulinus globosus, (Gastropoda: Planorbidae) from selected habitats of KwaZulu-Natal, South Africa.

The freshwater snail Bulinus globosus is an important intermediate host of Schistosoma haematobium, the causative agent of urinary schistosomiasis. This disease is of major health concern, especially in Africa where the majority of cases have been reported. In this study the inter- and intra-genetic diversity and population genetic structure of B. globosus from nine locations in the KwaZulu-Natal province of South Africa was studied using four polymorphic microsatellite loci (BgZ1-BgZ4). Moderate genetic diversity was detected within populations with a mean diversity (HE) of 0.49±0.09. The majority of populations significantly deviated from Hardy-Weinberg equilibrium (p<0.05), due to a deficit of heterozygotes. Such deviations may be due to founder events that were caused by bottlenecks that occurred as a result of frequent droughts and flooding that these snails' habitats are exposed to. Overall, the populations studied seem to be partially inbreeders/selfers with mean estimates of 0.24/0.38. A discernable genetic structure was elucidated among populations as evident by the mean pairwise FST of 0.58±0.13. There was no significant association between genetic and geographical distance among populations, an indication of limited gene flow. This increases the chances of populations losing alleles due to genetic drift. Populations in close proximity demonstrated high genetic differentiation (58.77% total variation) due to allelic differences between them. The sample populations fell into 12 clusters, however, the populations from uMkhanyakude and uThungulu exhibited no discernable genetic structure. Genetically, the Bhobhoyi site found within the uGu district was equidistant to the two main sampling regions.

Phylogeny of seven Bulinus species originating from endemic areas in three African countries, in relation to the human blood fluke Schistosoma haematobium.

BACKGROUND: Snails species belonging to the genus Bulinus (Planorbidae) serve as intermediate host for flukes belonging to the genus Schistosoma (Digenea, Platyhelminthes). Despite its importance in the transmission of these parasites, the evolutionary history of this genus is still obscure. In the present study, we used the partial mitochondrial cytochrome oxidase subunit I (cox1) gene, and the nuclear ribosomal ITS, 18S and 28S genes to investigate the haplotype diversity and phylogeny of seven Bulinus species originating from three endemic countries in Africa (Cameroon, Senegal and Egypt). RESULTS: The cox1 region showed much more variation than the ribosomal markers within Bulinus sequences. High levels of genetic diversity were detected at all loci in the seven studied species, with clear segregation between individuals and appearance of different haplotypes, even within same species from the same locality. Sequences clustered into two lineages; (A) groups Bulinus truncatus, B. tropicus, B. globosus and B. umbilicatus; while (B) groups B. forskalii, B. senegalensis and B. camerunensis. Interesting patterns emerge regarding schistosome susceptibility: Bulinus species with lower genetic diversity are predicted to have higher infection prevalence than those with greater diversity in host susceptibility. CONCLUSION: The results reported in this study are very important since a detailed understanding of the population genetic structure of Bulinus is essential to understand the epidemiology of many schistosome parasites.

Significant population genetic structure of the Cameroonian fresh water snail, Bulinus globosus, (Gastropoda: Planorbidae) revealed by nuclear microsatellite loci analysis.

In order to characterize the demographic traits and spatial structure of Cameroonians Bulinus globosus, intermediate host of Schistosoma haematobium, genetic structure of seven different populations, collected from the tropical zone, was studied using six polymorphic microsatellites. Intrapopulation genetic diversity ranged from 0.37 to 0.55. Interpopulation genetic diversity variation clearly illustrated their significant isolation due to distance with gene flow substantially limited to neighbouring populations. The effective population sizes (Ne) were relatively low (from 3.0 to 18.6), which supposes a high rate from which populations would lose their genetic diversity by drift. Analysis of genetic temporal variability indicated fluctuations of allelic frequencies (35 of 42 locus-population combinations, P<0.05) characteristic of stochastic demography, and this is reinforced by events of bottlenecks detected in all populations. These findings demonstrated that Cameroonian B. globosus were mixed-maters with some populations showing clear preference for outcrossing. These data also suggest that genetic drift and gene flow are the main factors shaping the genetic structure of studied populations.

The ITS2 of the genus Bulinus: novel secondary structure among freshwater snails and potential new taxonomic markers.

The freshwater snail genus Bulinus has been intensively investigated due to its role as intermediate host for trematode blood flukes that cause the debilitating disease schistosomiasis in man and livestock. Owing to taxonomic ambiguities within Bulinus, attention has often focused upon species delineation and several molecular methods have recently been used for identification and characterization purposes. Inspection of compensatory base changes (CBCs) in the secondary structure of the nuclear ribosomal internal transcribed spacer (ITS) has been used to differentiate species in other genera, and here we present a study investigating the presence of CBCs between species in the species groups within Bulinus. CBCs were present within B. forskalii and B. globosus indicating that these widely distributed taxa might constitute cryptic species complexes. However, other currently recognized species could not be distinguished by CBC analysis. The putative secondary structure of the very long ITS2 sequence of the B. reticulatus species group had an additional helix (DIIa) between DII and DIII not seen in other species groups of Bulinus. The accumulation and inspection of further ITS2 sequences will no doubt reveal additional variation between Bulinus populations, and CBCs should be incorporated in future taxonomic work in this group.

Parasitological and malacological surveys reveal urogenital schistosomiasis on Mafia Island, Tanzania to be an imported infection.

To confirm the local endemicity of Schistosoma haematobium on Mafia Island, Tanzania, conjoint parasitological and malacological surveys were undertaken in July 2006 with parasitological investigations supplemented with case-history questionnaires. A total of 238 children (125 girls and 113 boys, mean age of 13.9 years) across 9 primary schools were examined. The prevalence of micro-haematuria and egg-patent infection was 18.1% (CI95=9.6-33.6) and 4.2% (CI95=1.9-7.6), respectively but a strong female bias was observed for micro-haematuria (5.6F:1M) contrasting with a strong male bias for the presence of eggs (1F:4M). All egg-patent infections were of light-intensity (<10eggs/10ml). No clear associations between infection prevalence and local water-contact, by school, were found and all 10 of the egg-positive children had a travel history to the nearby mainland or Zanzibar. Inspection of community diagnostic registers at Kilindoni Hospital revealed a low proportion (<2%) of egg-patent infection for 20,306 samples tested in the 2000-2005 period. A total of 43 freshwater sites, a third of which were previously sampled in 1999 and 2002, were surveyed and 11 species of freshwater mollusc were found. Four species of Bulinus (B. nasutus, B. forskalii, B. barthi and B. sp.) were encountered across 13 sites with B. nasutus restricted to 3 of these towards the north of the island. No collected snail was observed to shed schistosome cercariae. Further characterisation of B. nasutus and S. haematobium included infection challenge on two occasions, with miracidia obtained from egg-patent children from Mafia and Unguja islands as well as DNA barcoding of snails and schistosomes. B. nasutus was shown refractory to infection. With the substantial travel to and from Mafia, the refractory nature of local snails and evidence from DNA barcoding in schistosomes and snails, we conclude that urogenital schistosomiasis is an imported infection.

Bulinus globosus (Planorbidae; Gastropoda) populations in the Lake Victoria basin and coastal Kenya show extreme nuclear genetic differentiation.

Bulinus globosus, a key intermediate host for Schistosoma haematobium that causes urinary schistosomiasis, is a hermaphroditic freshwater Planorbid snail species that inhabits patchy and transient water bodies prone to large seasonal variations in water availability. Although capable of self-fertilizing, this species has been reported to be preferentially out crossing. In this study, we characterized the population genetic structure of 19 B. globosus populations sampled across the Lake Victoria basin and coastal Kenya using four polymorphic microsatellite loci. Population genetic structure was characterized and quantified using FST statistics and Bayesian clustering algorithms. The four loci used in this study contained sufficient statistical power to detect low levels of population genetic differentiation and were highly polymorphic with the number of alleles per locus across populations ranging from 16 to 22. Average observed and expected heterozygosities across loci in each population ranged from 0.13 to 0.69 and from 0.39 to 0.79, respectively. Twenty-five of the seventy-six possible population-locus comparisons significantly deviated from Hardy-Weinberg equilibrium proportions after Bonferroni corrections, mostly due to the deficiency of heterozygotes. Significant genetic differentiation was observed between populations and Bayesian inferences identified 15 genetic clusters. The excess homozygosity, significant inbreeding and population genetic differentiation observed in B. globosus populations are likely to be due to the habitat patchiness, mating system and the proneness to cyclic extinction and recolonization in transient habitats.

Malacological survey and geographical distribution of vector snails for schistosomiasis within informal settlements of Kisumu City, western Kenya.

BACKGROUND: Although schistosomiasis is generally considered a rural phenomenon, infections have been reported within urban settings. Based on observations of high prevalence of Schistosoma mansoni infection in schools within the informal settlements of Kisumu City, a follow-up malacological survey incorporating 81 sites within 6 informal settlements of the City was conducted to determine the presence of intermediate host snails and ascertain whether active transmission was occurring within these areas. METHODS: Surveyed sites were mapped using a geographical information system. Cercaria shedding was determined from snails and species of snails identified based on shell morphology. Vegetation cover and presence of algal mass at the sites was recorded, and the physico-chemical characteristics of the water including pH and temperature were determined using a pH meter with a glass electrode and a temperature probe. RESULTS: Out of 1,059 snails collected, 407 (38.4%) were putatively identified as Biomphalaria sudanica, 425 (40.1%) as Biomphalaria pfeifferi and 227 (21.5%) as Bulinus globosus. The spatial distribution of snails was clustered, with few sites accounting for most of the snails. The highest snail abundance was recorded in Nyamasaria (543 snails) followed by Nyalenda B (313 snails). As expected, the mean snail abundance was higher along the lakeshore (18 ± 12 snails) compared to inland sites (dams, rivers and springs) (11 ± 32 snails) (F(1, 79) = 38.8, P < 0.0001). Overall, 19 (1.8%) of the snails collected shed schistosome cercariae. Interestingly, the proportion of infected Biomphalaria snails was higher in the inland (2.7%) compared to the lakeshore sites (0.3%) (P = 0.0109). B. sudanica was more abundant in sites along the lakeshore whereas B. pfeifferi and B. globosus were more abundant in the inland sites. Biomphalaria and Bulinus snails were found at 16 and 11 out of the 56 inland sites, respectively. CONCLUSIONS: The high abundance of Biomphalaria and Bulinus spp. as well as observation of field-caught snails shedding cercariae confirmed that besides Lake Victoria, the local risk for schistosomiasis transmission exists within the informal settlements of Kisumu City. Prospective control interventions in these areas need to incorporate focal snail control to complement chemotherapy in reducing transmission.

Molecular approaches to the identification of Bulinus species in south-west Nigeria and observations on natural snail infections with schistosomes.

The current study considers the distribution of a small sample of 138 Bulinus snails, across 28 localities within eight Nigerian states. Snails were identified using a combination of molecular methods involving both DNA sequencing of a partial cytochrome oxidase subunit 1 (cox1) fragment and restriction profiles obtained from ribosomal internal transcribed spacer (its) amplicons. The results showed that the majority of Bulinus samples tested belonged to the species Bulinus truncatus while only two were Bulinus globosus. The use of RsaI restriction endonuclease to cleave the ribosomal its of Bulinus, as a method of species identification, was adopted for the majority of samples, this being a quicker and cheaper method better suited to small laboratory environments. Polymerase chain reaction (PCR) amplification of the schistosome Dra1 repeat within each of the collected Bulinus samples was employed to determine the extent and distribution of infected snails within the sample areas. Successful amplification of the Dra1 repeat demonstrated that 29.7% of snails were infected with schistosomes. Sequencing of the partial schistosome its from a small subset of snail samples suggested that some snails were either penetrated by both Schistosoma haematobium and Schistosoma bovis miracidia or hybrid miracidia formed from the two species.

Molecular phylogeny of diploid Bulinus sp. (Gastropoda: Planorbidae) populations in Cameroon crater lakes.

Bulinus sp. (2n=36) is a diploid freshwater snail found in Cameroon crater lakes; it belongs to a group of medically important freshwater snails. Some members (Bulinus truncatus, Bulinus tropicus) of this group had been reported to be involved in the transmission of parasites (Schistosoma sp. and Calicophoron microbothrium) to human and livestock in tropical Africa. Yet, understanding of the evolutionary identity of the diploid snail such as its phylogenetic position and the genetic divergence among populations, remains limited. In this study, we constructed the molecular phylogeny of Bulinus sp. using sequences of mitochondrial cytochrome oxydase subunit 1 (CO-1, 365 nucleotides). Partial sequences of CO-1 were obtained and genetic divergences between populations estimated after the alignment of 365 nucleotides from each studied population. The lack of deep molecular divergences between populations of Bulinus sp. from western Cameroon crater lakes may indicate that they belong to the same lineage; therefore, it implies that diploid B. truncatus/tropicus complex snail-like in Cameroon share a common ancestor. The CO-1 of the three studied populations of Bulinus sp., clustered together with other diploid pan-African representatives of the B. truncatus/tropicus complex, showed little evidence of genetic similarities.

Molluscicidal activity of some Cameroonian plants on Bulinus species.

OBJECTIVE: To evaluate some locally available plants for their molluscicidal activity on Bulinus camerunensis and B. truncatus (slender form). DESIGN: Experimental studies. SETTING: Ndongo stream near the University of Buea and the University of Buea Life Sciences Laboratory. SUBJECTS: Evaluation of molluscicidal activity on snails of Bulinus camerunensis and B. truncatus (slender form). MAIN OUTCOME MEASURES: Plant extracts with molluscicidal activity determined. Determination of LC50, LC90 and LC100 of the potent plant extracts. Application of the extracts on aquaria-reared snails. Semi-field application of extracts. RESULTS: A preliminary screening test using 10,000 ppm solution of the water extracts of thirteen plants revealed that 61.5% (8/13) of the plants investigated had molluscicidal properties, with snail mortality rates above 90%. Extracts of Nicotiana tabacum, Aframomum citratum, A. melegueta, Curcuma domestica and Solanum scabrum killed 100% of the snails after twenty four hours exposure. B. camerunensis was more susceptible to the water extracts than B. truncatus. The LC50, LC90 and LC100 of the different plant extracts against B. camerunensis were generally lower than those against B. truncatus. The concentrations that produced 50%, 90% and 100% snail mortalities were lower with the methanol extracts than with water extracts, indicating that the methanol extracts were more toxic. to the snails than the water extracts. Generally, the eggs were more susceptible to the extracts than the young and adult snails. Application of the water extracts at LC 100 on snails reared in aquaria and under semi-field conditions revealed that N. tabacum could kill up to 100% of the snails in aquaria and 61.25% under semi-field conditions. CONCLUSION: Eight plant species with molluscicidal activity were identified, among which Nicotiana tabacum, Aframomum citratum, A. melegueta, Solanum scabrum and Curcuma domestica presented the highest activity. B. camerunensis was more susceptible to all the plant extracts tested than B. truncatus, and the methanol extracts proved more toxic than the water extracts. Semi-field testing of potent extracts showed promise, with N. tabacum having the highest effects on the snails.

Spot light survey on fresh-water snails of medical importance in Al Fayoum Governorate, Egypt.

In a survey carried out during Summer and Autumn of 2004, for snails of medical importance, nine species were recovered. These were Biomphalaria alexandrina, B. glabrata, B. pfeifferi, Bulinus truncatus, B. forskalii, Lymnaea natalensis, Bellamya (=Vivipara) unicolor, Physa acuta and Hydrobia musaensis. Parasitological examination revealed that B. alexandrina, B. glabrata and L. natalensis harboured immature stages of their concerned trematode parasites. Moreover, P. acuta harboured the immature stage of the nematode parasite Parastrongylus cantonensis.

Breeding system and demography shape population genetic structure across ecological and climatic zones in the African freshwater snail, Bulinus forskalii (Gastropoda, Pulmonata), intermediate host for schistosomes.

The role of breeding system and population bottlenecks in shaping the distribution of neutral genetic variation among populations inhabiting patchily distributed, ephemeral water bodies was examined for the hermaphroditic freshwater snail Bulinus forskalii, intermediate host for the medically important trematode Schistosoma guineensis. Levels of genetic variation at 11 microsatellite loci were assessed for 600 individuals sampled from 19 populations that span three ecological and climatic zones (ecozones) in Cameroon, West Africa. Significant heterozygote deficiencies and linkage disequilibria indicated very high selfing rates in these populations. Despite this and the large genetic differentiation detected between populations, high levels of genetic variation were harboured within these populations. The high level of gene flow inferred from assignment tests may be responsible for this pattern. Indeed, metapopulation dynamics, including high levels of gene flow as well as extinction/contraction and recolonization events, are invoked to account for the observed population structuring, which was not a consequence of isolation-by-distance. Because B. forskalii populations inhabiting the northern, Sahelian area are subject to more pronounced annual cycles of drought and flood than the southern equatorial ones, they were expected to be subject to population bottlenecks of increased frequency and severity and, therefore, show reduced genetic variability and elevated population differentiation. Contrary to predictions, the populations inhabiting the most northerly ecozone exhibited higher genetic diversity and lower genetic differentiation than those in the most southerly one, suggesting that elevated gene flow in this region is counteracting genetic drift.

[Experimental study of the compatibility between Schistosoma haematobium and two species of Bulinus in Cameroon]. / Etude expérimentale de la compatibilité entre Schistosoma haematobium et deux espèces de bulins au Cameroun.

A study on the compatibility between Schistosoma haematobium from three remote localities (Mourtourwa, Gounougou and Kékem) and four populations of Bulinus truncatus (Gounougou, Ngaoundéré, Bertoua and Kékem) and four populations of B. globosus (Mourtourwa, Ouroudoukoudje, Bafia and Yaoundé) was undertaken in order to estimate the risk of extension of urinary schistosomiasis in Cameroon. First generation of offspring from wild Bulinus was exposed to miracidia liberated by schistosome eggs extracted from patient urine. Between the 25th and the 60th day post-infestation the number of snails still alive, the number emitting cercariae and the prepatent period duration were noted. Results showed that all B. truncatus samples were susceptible to the three strains of parasite whereas only B. globosus of Mourtourwa and Ouroudoukoudje were susceptible to S. haematobium from Mourtourwa. The schistosome infection rate was then significantly higher in B. truncatus and the prepatent period significantly lower than in B. globosus. The compatibility characterised by a high infection rate and a low prepatent period was significantly better in homopatric couples than in allopatric combinations. The results suggested that B. truncatus might be potentially more implicated than B. globosus to the extension of the urinary bilharziasis in Cameroon.

Miracidia of an Egyptian strain of Schistosoma mansoni differentiate between sympatric snail species.

The host-finding behavior of miracidia of 2 strains of Schistosoma mansoni from Egypt and Brazil was studied by recording their responses to snail-conditioned water (SCW) from the Egyptian sympatric snails, Biomphalaria alexandrina, Physa acuta, Lymnaea cailliudi, and Balinus truncatus, as well as from Biomphalaria arabica and Biomphalaria glabrata. Miracidia of the Egyptian strain significantly preferred SCW from their compatible hosts B. alexandrina and B. arabica and showed no or a weak response to SCW from the other sympatric species, whereas miracidia of the Brazilian strain did not differentiate between SCW from different snail species.

A comparison of Bulinus africanus group species (Planorbidae; Gastropoda) by use of the internal transcribed spacer 1 region combined by morphological and anatomical characters.

The morphological and anatomical characters applied to determine species identity in the Bulinus africanus group species are insufficient to unambiguously discriminate between arbitrary species of given populations. In order to solve this problem, four snail populations from Kenya have been investigated morphologically and anatomically, and the species status compared with the result of molecular methods. We have amplified the entire ITS region and found that the investigated populations showed intra-specific genetic polymorphism, thereby giving the taxa an identity which were indistinct when the region was cut with restriction enzymes. Instead, an amplification of the sub-region ITS 1 revealed an unambiguous identification. because the amplification revealed only one single fragment. We also found that the observed heterogeneity of the entire ITS region could be confined to the sub-region ITS 2. Furthermore, the micro-sculpture of the shell and penis to preputium proportion, which are normally applied as morphological characters, might be considered as inadequate, because of the lack of a significant difference in those characters between the two well established species, namely B. africanus and B. nasutus in the Kisumu area. Instead, these two taxa were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to be one single species with a highly variable morphology. This result was further confirmed by random amplified polymorphic DNA (RAPD) data and the mitochondrial cytochome oxidase subunit I (COI).

Variation of microsatellite size homoplasy across electromorphs, loci, and populations in three invertebrate species.

Size homoplasy was analyzed at microsatellite loci by sequencing electromorphs, that is, variants of the same size (base pairs). This study was conducted using five interrupted and/or compound loci in three invertebrate species, the honey bee Apis mellifera, the bumble bee Bombus terrestris, and the freshwater snail Bulinus truncatus. The 15 electromorphs sequenced turned out to hide 31 alleles (i.e., variants identical in sequence). Variation in the amount of size homoplasy was detected among electromorphs and loci. From one to seven alleles were detected per electromorph, and one locus did not show any size homoplasy in both bee species. The amount of size homoplasy was related to the sequencing effort, since the number of alleles was correlated with the number of copies of electromorphs sequenced, but also with the molecular structure of the core sequence at each locus. Size homoplasy within populations was detected only three times, meaning that size homoplasy was detected mostly among populations. We analyzed population structure, estimating Fst and a genetic distance, based on either electromorphs or alleles. Whereas little difference was found in A. mellifera, uncovering size homoplasy led to a more marked population structure in B. terrestris and B. truncatus. We also showed in A. mellifera that the detection of size homoplasy may alter phylogenetic reconstructions.