Schistosomes of small mammals from the Lake Victoria Basin, Kenya: new species, familiar species, and implications for schistosomiasis control.

Recent schistosomiasis control efforts in sub-Saharan Africa have focused nearly exclusively on treatment of humans with praziquantel. However, the extent to which wild mammals act as reservoirs for Schistosoma mansoni and therefore as sources of renewed transmission following control efforts is poorly understood. With the objective to study the role of small mammals as reservoir hosts, 480 animals belonging to 9 rodent and 1 insectivore species were examined for infection with schistosomes in Kisumu, in the Lake Victoria Basin, Kenya. Animals were collected from 2 sites: near the lakeshore and from Nyabera Marsh draining into the lake. A total of 6.0% of the animals captured, including 5 murid rodent species and 1 species of shrew (Crocidura olivieri) were infected with schistosomes. Four schistosome species were recovered and identified using cox1 DNA barcoding: S. mansoni, S. bovis, S. rodhaini and S. kisumuensis, the latter of which was recently described from Nyabera Marsh. Schistosoma mansoni and S. rodhaini were found infecting the same host individual (Lophuromys flavopunctatus), suggesting that this host species could be responsible for the production of hybrid schistosomes found in the area. Although the prevalence of S. mansoni infection in these reservoir populations was low (1.5%), given their potentially vast population size, their impact on transmission needs further study. Reservoir hosts could perpetuate snail infections and favour renewed transmission to humans once control programmes have ceased.

Schistosoma kisumuensis n. sp. (Digenea: Schistosomatidae) from murid rodents in the Lake Victoria Basin, Kenya and its phylogenetic position within the S. haematobium species group.

Schistosoma kisumuensis n. sp. is described based on 6 adult males and 2 adult females collected from the circulatory system of 3 murid rodent species, Pelomys isseli, Mastomys natalensis, and Dasymys incomtus. Specimens were collected from a single location, Nyabera Swamp, in Kisumu, Kenya in the Lake Victoria Basin. This new species is morphologically similar to members of the S. haematobium group, currently represented by 8 species parasitizing artiodactyls and primates, including humans. Schistosoma kisumuensis differs from these species by producing relatively small Schistosoma intercalatum-like eggs (135.2 x 52.9 microm) with a relatively small length to width ratio (2.55). Comparison of approximately 3000-base-pair sequences of nuclear rDNA (partial 28S) and mtDNA (partial cox1, nad6, 12S) strongly supports the status of S. kisumuensis as a new species and as a sister species of S. intercalatum. The cox1 genetic distance between these two species (6.3%) is comparable to other pairwise comparisons within the S. haematobium group. Separation of the Congo River and Lake Victoria drainage basins is discussed as a possible factor favoring the origin of this species.

Microsatellite typing reveals strong genetic structure of Schistosoma mansoni from localities in Kenya.

Genetic diversity and population structure of seven populations of Schistosoma mansoni sampled in Kenya were assessed using five microsatellite markers. The mean number of alleles per locus, expected heterozygosity in Hardy-Weinberg equilibrium and pairwise F(ST) values ranged from 5.2 to 10.7, 0.5-0.8 and 3.6-27.3%, respectively. These data reveal that S. mansoni populations in Kenyan have relatively high levels of genetic diversity and is significantly differentiated. Our data combined with information on biogeography support the hypothesis that the strong genetic structure in Kenyan schistosomes is as a result of limited gene flow and large population sizes. Resistance to anthelminthics has not been reported among the Kenyan schistosomes, we hypothesize that this is probably due to the very little gene flow among populations, thereby limiting opportunities for the spread of rare alleles that might confer resistance to the drugs.

Impact of the crayfish Procambarus clarkii on Schistosoma haematobium transmission in Kenya.

The Louisiana red swamp crayfish, Procambarus clarkii, which was introduced into east Africa in the 1950s or 1960s, has since widely dispersed. Previous work by our group has shown that P. clarkii can reduce populations of the molluscan intermediate hosts of human schistosomes through predatory and competitive interactions. Here, we investigate whether crayfish can reduce populations of Bulinus africanus and consequently, Schistosoma haematobium prevalence in school children. Children from 6 primary schools in the Machakos and Kitui Districts of Kenya were selected for study. Schools were divided into 3 experimental-control pairs. At experimental schools, crayfish were introduced into nearby aquatic habitats harboring Bulinus africanus snails and serving as S. haematobium transmission sites. Snail habitats near control schools did not receive crayfish. Six months after crayfish introduction, all infected children were treated with praziquantel. Children were then monitored quarterly for 2 years, at which time infection and reinfection rates were compared statistically between the paired schools. In one such pair, crayfish failed to establish, resulting in neither snail control nor a reduction in transmission. At the second pair of schools, the numbers of snails were decreased by the presence of crayfish, but a clear difference in infection rates in children could not be detected, primarily because drought conditions kept overall transmission rates low. At the third school pair, crayfish established well in experimental habitats, snail numbers decreased precipitously, and children at the experimental school were significantly less likely to acquire S. haematobium infections than children at the control school. Our results indicate that under certain environmental circumstances, P. clarkii exerts a significant impact on the transmission of human schistosomiasis in Kenya. Important questions remain regarding the impact of P. clarkii on Kenyan freshwater ecosystems, not the least of which is its potential to significantly influence the epidemiology of schistosomiasis in east Africa.

Predation of aquatic stages of Anopheles gambiae by the Louisiana red swamp crawfish (Procambarus clarkii).

Laboratory experiments lasting 1-3 days were conducted in 10-liter glass aquaria to evaluate the ability of the Louisiana red swamp crawfish (Procambarus clarkii [Decapoda: Cambaridae]) to consume the aquatic forms of mosquitoes. With Anopheles gambiae as the target species, significantly fewer mosquito larvae or pupae survived in the presence of either juvenile or adult crawfish, relative to conditions without crawfish, regardless of whether crawfish had alternative food or not. When alternative food was excluded, juvenile and adult crawfish had a comparable ability to consume mosquito larvae. However, when alternative food was available, adult crawfish consumed significantly fewer mosquito larvae than did juveniles. In the case of pupae, juvenile crawfish consumed significantly more mosquito pupae than did the adults when alternative food was excluded. No significant difference, relative to controls, was found in the proportion of mosquito pupae surviving when adult crawfish had alternative food. Results of the present study show that P. clarkii has the ability to consume the aquatic forms of anopheline mosquitoes and, therefore, may have an impact on populations of pathogen-transmitting mosquitoes in an area of Kenya where the crawfish has become common.

Genetic diversity and population structure of Schistosoma mansoni within human infrapopulations in Mwea, central Kenya assessed by microsatellite markers.

A recently developed high-throughput technique that allows multi-locus microsatellite analysis of individual miracidia of Schistosoma mansoni was used to assess the levels of genetic diversity and population structure in 12 infrapopulations of the parasite, each infrapopulation derived from an infected school child from the Mwea area, central Kenya. The mean number of alleles per locus was in the range 8.22-10.22, expected heterozygosity in Hardy-Weinberg equilibrium was 0.68-0.70, and pairwise F(ST) values ranged from 0.16% to 3.98% for the 12 infrapopulations. Although the genetic diversity within each infrapopulation of S. mansoni in this area was generally high, low levels of genetic structure were observed, suggestive of high levels of gene flow among infrapopulations. Private alleles were found in 8 of the 12 infrapopulation, the highest number of private alleles recorded per infrapopulation was 3. Our data suggest that the level of gene flow among infrapopulations of S. mansoni in Mwea is extremely high, thus providing opportunity for spread of rare alleles, including those that may confer character traits such as drug resistance and virulence.

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

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

Molluscicidal activity of Solanum aculeatum (family: Solanaceae) berries against Biomphalaria pfeifferi, Bulinus globosus and Lymnaea natalensis.

Aqueous suspensions of powder of sun- or freeze-dried berries of the plant Solanum aculeatum (Family Solanaceae), indigenous in Kenya, were tested for molluscicidal activity against Biomphalaria pfeifferi, Bulinus globosus and Lymnaea natalensis under laboratory conditions. One hundred or 50 mg powder L-1 of sun- or freeze-dried berries killed over 60% of the test B. pfeifferi, Bul. globosus and L. natalensis. Whereas 25 mg L-1 of the sun dried material killed less than 60% of the test snails, similar concentrations of the freeze dried molluscicide produced 60-80% mortality in the snails, under similar conditions. Using L. natalensis as the target snail, it was shown that the freeze dried material was more potent than the freeze-dried berries of S. incanum, S. nigrum or leaves of Polygonum senegalensis (Family Polygonaceae), all present in Kenya, and known to possess molluscicidal properties. The powdered material retained molluscicidal activity even after several months storage at room temperature. These findings suggest that S. aculeatum is a potent plant molluscicide, and has the potential for the control of vectors of schistosomiasis and fascioliasis in Kenya.