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The genetical and environmental determination of phally polymorphism in the freshwater snail Bulinus truncatus.

In some species of self-fertile pulmonate snails, two sexual morphs co-occur in natural populations: regular individuals and aphallic individuals that cannot transmit sperm to other snails. Purely aphallic populations therefore reproduce obligatorily by selfing. Understanding the evolution of aphally and selfing in these snails requires a precise knowledge of phally determination. In this paper, we investigate the genetic and environmental determination of aphally in Bulinus truncatus by a survey of the family (offspring) aphally ratio of 233 individuals originating from seven natural populations and a study of the reaction norm of the family aphally ratio to temperature using 60 individuals from 10 selfed lineages of one population. Our results indicate a high genetic variability for the determination of aphally between populations and within some populations, associated with a high level of genetic determination. Our second experiment indicates a significant temperature and lineage effect though no interaction between these two effects. We discuss our results in the framework of threshold models developed for dimorphic traits with polygenic inheritance. We propose that the sexual morph of an individual at a given temperature is determined by a temperature threshold value depending on both the individual genotype and probabilistic processes.

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Microsatellites and the genetics of highly selfing populations in the freshwater snail Bulinus truncatus.

Hermaphrodite tropical freshwater snails provide a good opportunity to study the effects of mating system and genetic drift on population genetic structure because they are self-fertile and they occupy transient patchily distributed habitats (ponds). Up to now the lack of detectable allozyme polymorphism prevented any intrapopulation studies. In this paper, we examine the consequences of selfing and bottlenecks on genetic polymorphism using microsatellite markers in 14 natural populations (under a hierarchical sampling design) of the hermaphrodite freshwater snail Bulinus truncatus. These population genetics data allowed us to discuss the currently available mutation models for microsatellite sequences. Microsatellite markers revealed an unexpectedly high levels of genetic variation with < or = 41 alleles for one locus and gene diversity of 0.20-0.75 among populations. The values of any estimator of Fis indicate high selfing rates in all populations. Linkage disequilibria observed at all loci for some populations may also indicate high levels of inbreeding. The large extent of genetic differentiation measured by Fst, Rst or by a test for homogeneity between genic distributions is explained by both selfing and bottlenecks. Despite a limited gene flow, migration events could be detected when comparing different populations within ponds.

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Population dynamics inferred from temporal variation at microsatellite loci in the selfing snail Bulinus truncatus.

We analyzed short-term forces acting on the genetics of subdivided populations based on a temporal survey of the microsatellite variability in the hermaphrodite freshwater snail Bulinus truncatus. This species inhabits temporary habitats, has a short generation time and exhibits variable rates of selfing. We studied the variability over three sampling dates in 12 Sahelian populations (1161 individuals). Classical genetic parameters (estimators of Ho, He, f, selfing rate and Fst) showed limited change over time whereas important temporal changes of allelic frequencies were detected for 10 of the ponds studied. These variations are not easily explained by selection, sampling drift and genetic drift alone and may be due to periodic migration. Indeed the habitats occupied by the populations studied are subject to large temporal fluctuations owing to annual cycles of drought and flood. In such ponds our results support a demographic model of population expansions and contractions under which available habitats, after the rainy season, are colonized by individuals originating from a smaller number of refuges (areas that never dry out in the deepest parts of the ponds). In contrast, selfing appeared to be an important force affecting the genetic structure in permanent ponds.

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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.

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Comparative karyology of the freshwater snails Bulinus tropicus and B. natalensis.

Species of the subgenus Bulinus s.s. occur in a polyploid series which is of interest because of the rarity of polyploidy in animal species and because there appears to be a relation between ploidy level and ability to transmit Schistosoma haematobium, a parasite of major public health importance. We describe a technique for the preparation of standard karyotypes using cell suspensions from colchicine-treated embryos. The results for two diploid species, B. tropicus (Krauss) and B. natalensis (Küster), are presented. Both karyotypes consist of 15 metacentric and 3 submetacentric chromosomes. However, differences in centromere position are found in four pairs of chromosomes, and in the appearance of a secondary constriction. Because B. natalensis may represent a relict taxon involved in the origin of the polyploids, the results of significance for future work on the phylogeny of this group.

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The schistosome intermediate host, Bulinus nyassanus, is a 'preferred' food for the cichlid fish, Trematocranus placodon, at Cape Maclear, Lake Malawi.

Over the last decade, transmission of the schistosome parasites of humans has increased in parts of Lake Malawi, possibly because over-fishing has led to a decline in the numbers of molluscivorous fish. The stomach contents of 51 wild-caught Trematocranus placodon from the Cape Maclear area of Lake Malawi have now been investigated and compared with the field abundance of snail species at two sites, close to where the fish were caught. The snails found in the fish stomachs were identified to genus or species on the basis of their shell fragments and/or opercula. The sizes of the prosobranchs consumed by the T. placodon were determined from their operculum sizes whereas the Bulinus nyassanus ingested by the fish were categorized as small (<4 mm) or large (> or =4 mm) on the basis of their shell fragments. The proportion of the Bu. nyassanus ingested that were categorized as large increased with fish size. Furthermore, the relative abundance of Bu. nyassanus in the guts of the fish was considerably higher than in the field, indicating that the T. placodon had a 'preference' for Bu. nyassanus over the Melanoides species that dominated the snail fauna. Small specimens of Bellamya species were also consumed in a greater proportion than found in field.

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Identification of snails within the Bulinus africanus group from East Africa by multiplex SNaPshot trade mark analysis of single nucleotide polymorphisms within the cytochrome oxidase subunit I.

Identification of populations of Bulinus nasutus and B. globosus from East Africa is unreliable using characters of the shell. In this paper, a molecular method of identification is presented for each species based on DNA sequence variation within the mitochondrial cytochrome oxidase subunit I (COI) as detected by a novel multiplexed SNaPshotTM assay. In total, snails from 7 localities from coastal Kenya were typed using this assay and variation within shell morphology was compared to reference material from Zanzibar. Four locations were found to contain B. nasutus and 2 locations were found to contain B. globosus. A mixed population containing both B. nasutus and B. globosus was found at Kinango. Morphometric variation between samples was considerable and UPGMA cluster analysis failed to differentiate species. The multiplex SNaPshotTM assay is an important development for more precise methods of identification of B. africanus group snails. The assay could be further broadened for identification of other snail intermediate host species.

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Distribution, habitats and role as intermediate host of the freshwater snail, Bulinus forskalii, in South Africa.

This paper focuses on the geographical distribution and habitats of Bulinus forskalii, the snail intermediate host of the conical fluke of equids, Gastrodiscus aegyptiacus as reflected by the 1209 samples in the database of the National Freshwater Snail Collection of South Africa. The 362 different loci on record represent an extensive distribution in KwaZulu-Natal Province, the Limpopo Province, the coastal areas of the Eastern Cape Province and the south-eastern part of the North West Province. Although it was recorded from all types of water-body represented in the database, the highest percentages of samples were recovered from dams (30.4%) and brooks (28.2%). The majority of samples came from perennial habitats (59.1%), 60.7% from habitats with standing water, 54.0% from habitats with clear water and 71.8% from habitats of which the water was described as fresh. The majority of samples (39.5%) were collected in habitats of which the substratum was recorded as muddy. The highest percentage of samples, by far (81.5%), was collected in habitats that fell within the mean yearly temperature interval ranging from 15-20 degrees C. An integrated decision tree constructed from the data in the database indicated that temperature and type of water-body played a decisive role in determining the presence of B. forskalii in a given area. The results of experimental exposure to miracidia of a local strain of both Schistosoma haematobium and Schistosoma mattheei in the laboratory indicated that a local strain of B. forskalii was incompatible with both these strains of parasite. Research to clarify the role of B. forskalii in the transmission of both Calicophoron microbothrium and G. aegyptiacus in South Africa, is recommended.

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Effect of mass chemotherapy and piped water on numbers of Schistosoma haematobium and prevalence in Bulinus globosus in Kwale, Kenya.

From June 1982 to May 1986 in a small village in Kwale, Kenya, we studied seasonal fluctuations in populations of Bulinus globosus, prevalence of Schistosoma haematobium infection in this snail, and effects of chemotherapy and piped water supply on infection rate of snails. In the perennially-flowing Pemba River, relatively small numbers of snails were collected; they were found only during the hot dry season (December to March). In a tributary stream, the Kadingo River, whose flow ceased at the end of both the cool and hot dry seasons, snail numbers peaked at the end of the cool dry season (October to November) and at the beginning of the hot dry season (January). Large numbers of infected snails were found in the Kadingo River from November to January (short rainy season and beginning of dry season). Selective mass chemotherapy with metrifonate and provision of piped water were begun in February and March 1984. These control measures achieved a significant reduction in the infection rate of snails (P less than 0.001); the annual infection rate for the 2 years before treatment was 9.3% and 13.1%, and for the 2 years after treatment was 3.5% and 3.4%.

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A longifurcate distome cercaria from Bulinus truncatus snails in the Jordan Valley, Jordan.

Bulinus truncatus snails collected from water bodies of the South Shuna region, north of the Dead Sea, were found infected with a pharyngeal longifurcate distome cercaria. This new form of cercaria is named Cercaria bulini I. It has three pairs of penetration glands, 12 pairs of flame cells, and lacks the intestinal cecae. This cercaria is an active swimmer and develops within an elongated sporocyst. Other details on the morphology, behavior, and development of C. bulini I are presented.

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Techniques for estimating densities of Bulinus truncatus rohlfsi and its horizontal distribution in Volta Lake, Ghana.

Bulinus truncatus rohlfsi is the intermediate host of urinary schistosomiasis, which is highly endemic in the man-made Volta Lake. In 1971, a WHO/UNDP schistosomiasis project was established in the Pawmpawm branch and part of the Afram branch of Volta Lake. Malacological findings of the preliminary phase indicated that the snails were distributed in the littoral zone of the lake, and that this distribution was correlated with the presence of vegetation, especially Ceratophyllum. Transmission nearly always occurred in "water contact sites", i.e., places where people come into contact with water.A snail-sampling technique with palm-leaf mats was developed and standardized after it had been shown in sensitivity trials to compare favourably with a modified version of the "man-time" sampling method, in which the number of snails collected per man-hour is recorded. It is recommended that both these methods should be used to conduct ecological studies of B. rohlfsi in water contact sites.

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Laboratory assessment of the molluscicidal activity of Commiphora molmol (Myrrh) on Biomphalaria alexandrina, Bulinus truncatus and Lymnaea cailliaudi.

The molluscicidal properties of the oil extract of Commiphora molmol (Myrrh) were tested against Egyptian snail species: Biomphalaria alexandrina, Bulinus truncatus and Limnaea cailliaudi. The impact of the extract on the egg cluches of B. alexandrina and L. cailliaudi was also evaluated. Snails and their eggs were exposed for 24 and 48 hr at 22-26 degrees C to various concentrations of the extract. The results showed different susceptibilities B. alexandrina showed higher LD50 and LD90 (155, 195 ppm) than B. truncatus (50, 95 ppm) and L. cailliaudi (50, 85 ppm) after 24 hr exposure. 100% mortality was obtained for the egg cluches of B. alexandrina and L. cailliaudi at concentrations of 100 ppm and 75 ppm respectively. Lower concentrations were needed to obtain the same results after 48 hr. The present laboratory studies demonstrated that Myrrh has a molluscicidal effect on the snail intermediate hosts, particularly on their eggs. Field studies are recommended.

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Intermediate hosts of schistosoma; African Biomphalaria and Bulinus. I.

This study is an attempt to classify all described species of African Biomphalaria and Bulinus. It is based upon the author's examination of a great number of specimens collected from many parts of Africa. The variations attributable to age, environment and genetic factors which may be noted in the taxonomic characters are discussed, and some new species and subspecies are established. For each recognized species and subspecies the author states the distinguishing characters, indicates the geographical distribution, and gives a list of synonyms.

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Intermediate hosts of schistosoma: African Biomphalaria and Bulinus. II.

This study is an attempt to classify all described species of African Bulinus. It is based upon the author's examination of a great number of specimens collected from many parts of Africa. The variations attributable to age, environment and genetic factors which may be noted in the taxonomic characters are discussed, and some new species and subspecies are established. For each recognized species and subspecies the author states the distinguishing characters, indicates the geographical distribution, and gives a list of synonyms.

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Ecology and distribution of Bulinus truncatus in the Middle East; with comments on the effect of some human activities in their relationship to the snail host on the incidence of bilharziasis haematobia in the Middle East and Africa.

The author gives a comprehensive account of the present status of knowledge regarding the ecology and distribution of Bulinus truncatus, the intermediate host of Schistosoma haematobium in the Middle East and North Africa. He discusses in detail the types of habitat favoured by this snail, its life-cycle and seasonal variations in its populations, the effect of ecological factors on its distribution and occurrence, and the relationship between its control and its bionomics, stressing the many aspects of these subjects that require further investigation and suggesting lines for future research.In addition, the author outlines briefly the effect of some human activities (for example, extension of irrigation and agriculture), in their relationship to the snail host, on the incidence of bilharziasis haematobia in the Middle East and Africa. He concludes the paper with a summary of the three basic needs in respect of bilharziasis control: ecological research; prevention of pollution of waters with human excrement; and avoidance of human contact with infective water.

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[Bulinus truncatus, intermediate host of Schistosoma haematobium in the Senegal River Basin (SRB)].

To assess the role of Bulinus truncatus in the transmission of urinary schistosomiasis in the Senegal River Basin (SRB), the relations between B. truncatus and Schistosoma haematobium were studied. The compatibility study shows that B. truncatus is susceptible to infection with S. haematobium in the Upper Valley of the SRB. The malacological follow up reveals the presence of B. truncatus naturally infected with schistosomes cercariae in the Middle Valley. The identification of these schistosomes as S. haematobium by the Single Strand Conformational Polymorphism technique (SSCP) confirms the participation of B. truncatus in the dynamic transmission.

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Observations on mass production of Calicophoron microbothrium metacercariae from experimentally and naturally infected Bulinus tropicus.

In an attempt to establish an ideal method for mass production of Calicophoron microbothrium metacercariae, a study was carried out to compare the shedding capacities of Bulinus tropicus naturally and experimentally infected with C. microbothrium. A total of 906 F1 B. tropicus between 4 and 5 weeks old were each experimentally infected with two C. microbothrium miracidia and monitored for 12 weeks. The infected snails were fed on dried lettuce and fish flakes and were kept in 1 l plastic aquaria housed in a snail room where temperature, light and humidity were controlled. Seventy-four percent of the experimentally infected snails died during the prepatent period and of the remaining, only 13.2% developed patent infection, while 12.5% were refractory. Snail growth rate was poor and the average shedding rate was 20 cercariae per snail per day. Compared to the experimentally infected snails, 2200 adult B. tropicus, collected from the field and naturally infected with C. microbothrium, yielded high numbers of metacercariae. Eighty-four percent of the snails died within 7 weeks of the study with peak mortality occurring from the 2nd to the 4th week of infection and coinciding with an overall decrease in the number of cercariae shed.

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