Low genetic diversity in a snail intermediate host (Biomphalaria pfeifferi Krass, 1848) and schistosomiasis transmission in the Senegal River Basin.

Population genetic perturbations of intermediate hosts, often a consequence of human pressure on environmental resources, can precipitate unexpectedly severe disease outbreaks. Such disturbances are set to become increasingly common following range changes concomitant with climate shifts, dwindling natural resources and major infrastructure changes such as hydroprojects. Construction of the Diama dam in the Senegal River Basin (SRB) reduced river salinity, enabling the freshwater snail intermediate host Biomphalaria pfeifferi to rapidly expand its distribution. A serious public health problem ensued, with an epidemic of intestinal schistosomiasis occurring in the previously schistosome-free Richard-Toll region within 2 years. The current study aimed to assess the population variability of B. pfeifferi in the SRB, and speculate upon its subsequent impact on host-parasite interactions following such engineered ecological change. Genetic variation at nine polymorphic microsatellite loci revealed little population differentiation in SRB snails compared with those from natural habitats in Zimbabwe, where Schistosoma mansoni transmission is much lower. 'Open' SRB habitats are associated with greater water contact, smaller population sizes and less genetic diversity, with sites downstream of Richard-Toll showing greater inter- and intrapopulation variation, concomitant with less frequent human contact. These observations may be explained by rapid expansion into pristine habitat selecting for high fecundity genotypes at the expense of schistosome resistance, presenting S. mansoni with genetically homogenous highly fecund susceptible populations around the focal point, promoting development of a highly compatible host-parasite relationship. Longitudinal study of such systems may prove important in predicting public health risks engendered by future environmental engineering projects.

A single-strand conformation polymorphism (SSCP) approach for investigating genetic interactions of Schistosoma haematobium and Schistosoma guineensis in Loum, Cameroon.

Single-strand conformation polymorphism (SSCP) analysis of the second internal transcribed spacer (ITS2) of nuclear ribosomal DNA provides a molecular tool for the identification of Schistosoma haematobium, Schistosoma guineensis and the hybrids of these two species. This molecular tool was utilized to provide a detailed analysis of the interactions between S. haematobium and S. guineensis in hybrid zones of Loum, Littoral Province, Cameroon. Individual hybrid schistosomes were identified within the natural populations collected from Loum in 1990, 1999 and 2000, which would have been misidentified as S. haematobium using solely morphological and sequence criteria. This study indicates the complexities of the hybridization between S. haematobium and S. guineensis and emphasizes the importance of assessing morphological, biological and molecular data to gain insights into the interaction of these two species over time.

Towards control of schistosomiasis in sub-Saharan Africa.

Approximately 80% of the 200 million people infected with schistosomiasis inhabit sub-Saharan Africa, and the annual mortality is estimated to be 280,000. Praziquantel is the drug of choice in the treatment of schistosomiasis and pregnant women may now be treated. It was agreed at the World Health Assembly in 2001 that at least 75% of school-aged children in high burden areas should be treated for schistosomiasis and soil-transmitted helminth infections by 2010 to reduce morbidity. A grant from the Bill and Melinda Gates Foundation to the Schistosomiasis Control Initiative, Imperial College of Science, Technology and Medicine, London has enabled control programmes to be initiated in Uganda, Tanzania, Zambia, Burkina Faso, Niger and Mali. Additional programmes have recently commenced in Zanzibar with a grant from the Health Foundation to The Natural History Museum, London and in Cameroon. Combination treatment for schistosomiasis, gastrointestinal helminths and filariasis reduces costs of control programmes. The EC Concerted Action Group on 'Praziquantel: its central role in the chemotherapy of schistosome infection' met in Yaoundé Cameroon in 2004 to discuss recent developments in laboratory and field studies. The use of standard operating procedures will enable data on drug action on schistosomes produced in different laboratories to be compared. With the ever increasing use of praziquantel there is a possibility of the development of resistance by schistosomes to the drug, hence the necessity to explore the activities of other compounds. Artemether, unlike praziquantel, is effective against immature schistosomes. The effectiveness of mirazid, an extract of myrrh, is controversial as data from different laboratories are equivocal. It is suggested that an independent body such as the World Health Organization should determine whether mirazid should be used in the treatment of schistosomiasis.

The interaction of Schistosoma haematobium and S. guineensis in Cameroon.

Interactions between schistosomes are complex with some different species being able to mate and hybridize. The epidemiology of schistosomiasis in specific areas of South West Cameroon has evolved remarkably over 30 years as a result of hybridization between Schistosoma guineensis and S. haematobium. Morphological and biological data suggest that S. haematobium replaced S. guineensis in areas of Cameroon through introgressive hybridization. Data are reported on the use of single stranded conformational polymorphism (SSCP) analysis of the nuclear ribosomal second internal transcribed spacer (ITS2) of individual schistosomes from hybrid zones of Cameroon. The data show that since 1990 S. haematobium has completely replaced S. guineensis in Loum, with S. haematobium and the recombinants still present in 2000. This study illustrates the complexities of the dynamics between S. haematobium and S. guineensis in South West Cameroon.

[Bulinus truncatus, intermediate host of Schistosoma haematobium in the Senegal River Basin (SRB)]. / Bulinus truncatus, hôte intermédiaire de Schistosoma haematobium dans le bassin du fleuve Sénégal.

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.

Polyparasitism with Schistosoma haematobium and soil-transmitted helminth infections among school children in Loum, Cameroon.

Prevalence and abundance of Schistosoma haematobium and soil-transmitted helminths (STH) were assessed among a total of 1600 pupils (urine, n = 1190; faecal samples, n = 1454) attending five schools in Loum, Littoral Province, Cameroon, with the specific aim of assessing the extent of polyparasitism and the extent to which infections were focused in particular subsets of the study group. Prevalence of S. haematobium was 62.8% with an abundance (arithmetic mean of egg counts) of 54 eggs/10 ml urine. For the STH these were 47.7% and 619 eggs per gram of faeces (EPG) for Trichuris trichiura, 65.5% and 3636 EPG for Ascaris lumbricoides, and 1.4% and <0.1 EPG for hookworms. Most children (90.3%) were infected with at least one of these four species, the largest proportion (34.3%) carrying two species; 27.4% carried three and 1.1% carried concurrently all four species of parasites. The average number of species harboured increased with age, as did the prevalences of S. haematobium and T. trichiura but not that of A. lumbricoides. All STH showed marked differences in prevalence between the five schools but only T. trichiura varied significantly between sexes. Mean abundance of infection varied significantly between age classes, among schools and between the sexes, with females showing heavier mean EPGs for A. lumbricoides and T. trichiura and males higher mean eggs/10 ml urine for S. haematobium infections. A highly significant association was detected between A. lumbricoides and T. trichiura, that was not context-dependent. This was confirmed in quantitative analyses after controlling for differences in abundance between schools, sexes and age classes. A weaker context-dependent association (prevalence data) was detected between S. haematobium and A. lumbricoides (sex- and age-dependent) but quantitative associations between these two species, as well as between S. haematobium and T. trichiura, were not convincing.

Compatibility of Schistosoma haematobium, S. intercalatum and their hybrids with Bulinus truncatus and B. forskalii.

Schistosoma haematobium and S. intercalatum readily hybridize with each other producing generations of viable hybrid offspring. Experiments were designed to investigate the infectivity and viability of the S. haematobium x S. intercalatum F1 and F2 hybrid larvae in their two intermediate snail hosts compared with the parental species. Analysis of the data obtained suggested that the S. haematobium male x S. intercalatum female F1 hybrid miracidia were more infective to Bulinus truncatus than to B.forskalii, and also more infective to B. truncatus compared with the parental S. haematobium miracidia. This hybrid was also observed to have a greater cercarial productivity from both intermediate hosts and these cercariae were shown to be more infectious and to have a longer longevity compared with the cercariae of S. haematobium, S. intercalatum and the S. haematobium female x S. intercalatum male F1 hybrid cercariae. The S. haematobium female x S. intercalatum male F1 hybrid was shown not to be very successful in all stages of the investigations. The results indicate that the S. haematobium male x S. intercalatum female F1 hybrid may have many reproductive advantages over the reciprocal hybrid and the parental schistosome species. The significance of the results is discussed in relation to the epidemiological consequences occurring in Loum, Cameroon, and other areas where S. haematobium and S. intercalatum are sympatric and able to hybridize.

A phylogeny based on three mitochondrial genes supports the division of Schistosoma intercalatum into two separate species.

Two recognized strains of Schistosoma intercalatum, one from the Democratic Republic of Congo (DRC), formerly Zaire, and the other from Cameroon, have been investigated using DNA sequences from 3 mitochondrial genes, cytochrome oxidase subunit 1 (cox1), NADH dehydrogenase subunit 6 (nad6) and the small ribosomal RNA gene (rrnS). In addition, partial DNA sequences from the nuclear large subunit ribosomal RNA gene (lsrDNA) were included within the study. Although partial lsrDNA alone reveals little taxonomic information, phylogenetic analysis of the mitochondrial data demonstrates a clear dichotomy between the 2 purported strains and it is proposed that they should be treated as distinct taxa. The 'original' S. intercalatum now falls relatively basal in the S. haematobium group, while the proposed new species is more derived and sister taxon to S. bovis and S. curassoni.

Interactions between Schistosoma intercalatum (Zaire strain) and S. mansoni.

Schistosoma mansoni and S. intercalatum, two schistosomes from different evolutionary lineages, are parasitic in humans and therefore able to co-infect the same host where they occur sympatrically in Africa. Previous studies of mating interactions between these species in mice, using the Lower Guinea strain of S. intercalatum, have demonstrated the competitive dominance of S. mansoni over S. intercalatum in terms of pairing ability, which is potentially an important mechanism restricting the distribution of S. intercalatum in Africa. The study presented here examines the mating interactions in mice between S. mansoni and the Zaire (Democratic Republic of Congo) strain of S. intercalatum, which differs from the Lower Guinea strain in many biological characteristics. Analysis of the data showed a preponderance of intraspecific pairs over interspecific, demonstrating a specific mate preference system for both species. Mating competition between these species and the ability of males of both species to effect a change of mate by pulling paired females away from their partners was indicated. Comparisons are made between the competitive mating abilities of both strains of S. intercalatum relative to those of S. mansoni, with the data suggesting that S. mansoni is competitively dominant to S. intercalatum (Zaire) in sequential infections but to a lesser extent than for S. intercalatum (Lower Guinea). Additional factors which may contribute to the confinement of S. intercalatum (Zaire) to the Democratic Republic of Congo are discussed.

Isoenzyme analysis of Schistosoma haematobium, S. intercalatum and their hybrids and occurrences of natural hybridization in Cameroon.

Isoelectric focusing of glucose-6-phosphate dehydrogenase (G6PD) produced clearly identifiable profiles for S. haematobium and S. intercalatum and their hybrids. To provide a more detailed analysis of the interactions of S. haematobium and S. intercalatum in South West Cameroon over the last 12 years, G6PD analyses were carried out on individual schistosomes collected in Kumba in 1990, Loum in 1990, 1999 and 2000 and Barombi Mbo and Barombi Kotto in 1999. Studies were also carried out on the two parental species S. haematobium Barombi Mbo, S. intercalatum Edea and subsequent generations of hybrids resulting from laboratory crosses of the two parental species. The isoenzyme analysis demonstrated that the 1990 isolate from Kumba, was a recombinant of S. intercalatum x S. haematobium, and that 30% of individual schistosomes collected in 1990 in Loum were also recombinants. The remainder gave data indicative of S. haematobium. In 1999, 12.5% of individuals from Loum showed recombination and 10% in 2000. Results from the most recent parasitological survey in October 2000 showed the persistence of the recombinant population in addition to that of S. haematobium. There was also evidence of recombination having taken place in Barombi Kotto but not Barombi Mbo. This study demonstrates how the situation has changed over the last 12 years, and emphasizes the importance of assessing morphological, biological and molecular data together to gain a true picture of the rapidly evolving situation.

Mating interactions of Schistosoma haematobium and S. intercalatum with their hybrid offspring.

Experiments were designed to study the mating behaviour between the Schistosoma haematobium male x S. intercalatumfemale hybrid and the 2 parental species S. haematobium and S. intercalatum. Individual worms were identified by electrophoretic analysis of glucose-6-phosphate dehydrogenase, which was characteristic for each isolate. Analysis of the data obtained showed that both heterospecific and homospecific pairs formed between the hybrids and S. haematobium and S. intercalatum. S. haematobium and the hybrid are better than S. intercalatum in forming pairs, and S. haematobium showed a greater homospecific mate preference compared with the hybrid. Analysis of the data using the Mantel-Haenszel test suggests that mating competition does exist between the schistosomes, with the hybrid being dominant over both the parental species and S. haematobium being dominant over S. intercalatum. The hybrid males showed a greater ability than S. intercalatum and S. haematobium males in taking away S. haematobium and S. intercalatum females from their homospecific males when introduced into a pre-established S. haematobium or S. intercalatum infection. They were able to take females from S. intercalatum homospecific pairs more easily compared with females from S. haematobium homospecific pairs. The significance of the results is discussed in relation to the epidemiological changes of schistosomiasis in Cameroon, where hybridization between S. haematobium and S. intercalatum has taken place, with S. haematobium and the hybrid managing to replace the endemic S. intercalatum over the last 30 years.

The phylogeny of the Schistosomatidae based on three genes with emphasis on the interrelationships of Schistosoma Weinland, 1858.

Schistosomes are digenean flukes, parasitic of birds, mammals and crocodiles. The family Schistosomatidae contains species of considerable medical and veterinary importance, which cause the disease schistosomiasis. Previous studies, both morphological and molecular, which have provided a good deal of information on the phylogenetics of this group, have been limited in the number of species investigated or the type or extent of molecular data used. This paper presents the most comprehensive phylogeny to date, based on the sequences of 3 genes, complete ribosomal small subunit rRNA and large ribosomal subunit rRNA, and mitochondrial cytochrome oxidase 1, sequenced from 30 taxa including at least 1 representative from 10 of the 13 known genera of the Schistosomatidae and 17 of the 20 recognized Schistosoma species. The phylogeny is examined using morphological characters, intermediate and definitive host associations and biogeography. Theories as to the origins and spread of Schistosoma are also explored. The principal findings are that Ornithobilharzia and Austrobilharzia form a sister group to the Schistosoma; mammalian schistosomes appear paraphyletic and 2 Trichobilharzia species, T. ocellata and T. szidati, seem to be synonymous. The position of Orientobilharzia within the Schistosoma is confirmed, as is an Asian origin for the Schistosoma, followed by subsequent dispersal through India and Africa.

On the longevity of Schistosoma curassoni.

It is demonstrated that Schistosoma curassoni, a parasite of sheep, cattle and goats in parts of West Africa, will live for at least 8 years 5 months in a sheep. The sheep was exposed to 500 cercariae of S. curassoni liberated from infected Bulinus wrighti. The sheep died of natural causes, and at post-mortem 28 pairs of adult S. curassoni were removed from the mesenteric and rectal veins. All female worms were gravid, and eggs were hatched from faeces to produce miracidia. The development of immune responses of the host had apparently little or no effect on the viability of the eggs. Histological studies of the liver, small and large intestines revealed mild pathological symptoms. The longevity of S. curassoni is the first record of longevity of schistosomes to be based on worm counts.

Competitive mating interactions between Schistosoma haematobium and S. intercalatum (Lower Guinea strain).

Schistosoma haematobium and S. intercalatum belong to the S. haematobium group of schistosomes and can hybridize in nature where they are sympatric. They are therefore able to co-infect the same human host. Hybridization and competitive mating interactions with S. haematobium have been implicated in restricting the distribution of S. intercalatum in Africa and in the remarkably rapid replacement of S. intercalatum by S. haematobium at Loum, Cameroon. Previous studies have demonstrated the greater pairing ability of S. haematobium over S. intercalatum in hamsters infected with both species simultaneously or infected first with S. intercalatum (Lower Guinea strain) and later with S. haematobium. The present study demonstrates the greater pairing ability of S. haematobiumover S. intercalatum in hamsters infected first with S. intercalatum (Lower Guinea) and later with S. haematobium, and indicates that S. intercalatumis unlikely to spread to areas where S. haematobium is already established.

Mating interactions between Schistosoma mansoni and S. margrebowiei.

Previous studies of mating interactions between schistosome species from different evolutionary lineages in mice showed that mating is not random: heterospecific pairs form but, given the choice, the preference is for homospecific partners. Mating competition and change of mate were also demonstrated. Schistosoma mansoni is a medically important parasite of humans belonging to a different evolutionary group from S. margrebowiei, a parasite of bovines of veterinary importance. Experiments were designed to investigate the mating behaviour of these two species in mice. Analysis of the data showed a preponderance of intraspecific pairs over interspecific, demonstrating a specific mate preference system for both species. The existence of mating competition between these species was indicated. Males of both species were able to actively compete for paired females by pulling them away from their partners, thus effecting a change of mate. Overall, neither species appeared to be competitively dominant to the other, and the data suggest that where this is the case in sequential infections, the most successful species in terms of worm return and ability to form pairs will be the first species to infect the host.

The phylogeography of Asian Schistosoma (Trematoda: Schistosomatidae).

Partial (DNA) sequences are presented for 2 nuclear (18S and 28S rRNA genes) and 2 mitochondrial (12S rRNA and ND1 genes) loci for 5 species belonging to the Schistosoma japonicum, S. sinensium and S. indicum groups of Asian Schistosoma. Fresh field isolates were collected and cultured for the following taxa: S. incognitum (S. indicum group, central Thailand), S. mekongi (S. japonicum group, southern Laos), S. ovuncatum (S. sinensium group, northern Thailand), S. spindale (S. indicum group, northeast Thailand and central Thailand isolates) and S. sinensium (S. sinensium group, Sichuan Province, China). This represents the first published DNA sequence data for S. ovuncatum and for S. sinensium s.s. from the type locality in China. The paper also presents the first sequence data at the above loci for S. incognitum (except for the 28S sequences) and S. sinensium. Congruence was observed between the phylogenies estimated for each locus, although the relationships of S. incognitum were not so well resolved. Fitch-Margoliash, maximum likelihood (M/L) and maximum parsimony methods were used to estimate the phylogenies and the agreement between them was similar to that observed between loci. The ML tree was considered to best represent the data and additional 28S sequences (taken from the GenBank), for S. haematobium, S. japonicum, S. mansoni and Orientobilharzia turkestanicum, were used to construct an overall phylogeny. The S. indicum group taxa showed considerable divergence from the other Asian species and closest affinity with the African group. S. ovuncatum and S. sinensium appeared as sister taxa but their status as sibling species remained supported. The findings are discussed in the context of phylogeographical hypotheses for the origin of Schistosoma. An Asian origin for Schistosoma is also considered.

Application of single strand conformational polymorphism (SSCP) analysis with fluorescent primers for differentiation of Schistosoma haematobium group species.

To assess the utility of single-stranded conformational polymorphism (SSCP) analysis for the differentiation of schistosomes, using methods adapted for a Perkin Elmer ABI Prism 377 automated sequencer, 3 isolates of Schistosoma haematobium, 2 of S. intercalatum and single isolates of S. curassoni and S. bovis were selected for study. Two fluorescently labelled, double-stranded polymerase chain reaction products, amplified from the mitochondrial cytochrome oxidase subunit 1 (CO1) gene and the nuclear ribosomal second internal transcribed spacer (ITS2), were generated from single male and female worms. Changes in electrophoretic mobility of fragments within an SSCP profile revealed variation at individual, isolate and species levels. The mutational basis between representative SSCP profiles was confirmed by direct sequencing, demonstrating that single point substitutions were detectable. SSCP analysis has considerable potential as an alternative molecular method of identification and characterization of schistosomes. More broadly, fluorescence-based SSCP analysis is applicable to almost any gene target from any species of parasite and is a powerful molecular tool for genetic profiling.

Implication of Bulinus truncatus in the transmission of urinary schistosomiasis in Senegal, West Africa.

Studies on the compatibility between Bulinus truncatus and Schistosoma haematobium from various areas in the Senegal River basin (SRB) were carried out, to investigate the role of B. truncatus in the epidemiology of urinary schistosomiasis in Senegal. The results show that B. truncatus from the Lower Valley is not compatible with S. haematobium from the Middle Valley. Interestingly, the study reveals that B. truncatus from the Upper Valley is compatible with S. haematobium from the same area, and that S. haematobium from the Middle Valley is compatible with B. truncatus from the Lower Valley. Bulinus truncatus naturally infected with S. haematobium have been collected from the Upper Valley of the SRB. These results indicate that B. truncatus is involved in the urinary schistosomiasis in Senegal.

A model to explain the replacement of Schistosoma intercalatum by Schistosoma haematobium and the hybrid S. intercalatum x S. haematobium in areas of sympatry.

Numerous hypotheses have been postulated to explain the rapidly changing parasitological situation in Loum, Cameroon as a result of the interaction between Schistosoma haematobium and S. intercalatum. The aim of this study is to test the various hypotheses using a simple mathematical model, incorporating equal and unequal sex ratios of adult schistosomes, recombinations, and levels of compatibility with the intermediate molluscan hosts, B. forskalii and B. truncatus. The model assuming an equal sex ratio does not fit with the existing field data in that it predicts a continued presence of S. intercalatum, S. haematobium and the hybrids. The model assuming a sex bias in favour of males, which reflects the situation usually observed in schistosome populations, predicts the loss S. intercalatum which indeed concurs with the most recent data.