Transmission sites for Schistosoma haematobium and Schistosoma bovis identified in localities within the Athi River basin of Kenya using a PCR-RFLP assay

Background: The epidemiology of human urinary schistosomiasis caused by Schistosoma haematobium can be complicated by the presence of ruminant schistosomiasis caused, primarily by S. bovis. The two schistosome species may be transmitted by the same Bulinus species, they may occur sympatrically in the same habitat, and their cercariae are very similar in morphology and therefore, difficult to tell them apart. Screening of snails collected from freshwater habitats for schistosome infections is often used to identify transmission sites or to evaluate success or failure of interventions. However, pin-pointing sites involved in S. haematobium transmission can be complicated by the presence of other mammalian schistosomes such as the bovine schistosome, which is a fairly common parasite. A PCR-RFLP method targeting a unique segment of the second internal transcribed spacer (ITS2) region of the ribosomal DNA (rDNA) in the schistosomes was used to identify mammalian schistosome cercariae shed by bulinid snails collected from endemic freshwater habitats located within Machakos county in south-eastern Kenya, with the aim to identify the transmission sites and assess the distribution each of the parasite species in the study area. Results: A total of 5,034 bulinid snails were collected from 41 different sites and screened for schistosome infections, and out of these, 43 (<1%) were found to be shedding mammalian schistosome cercariae. On analysis using the Polymerase chain reactionRestriction Fragment Length Polymorphisms (PCR-RFLP) assay, cercariae from 32 snails were identified as S. haematobium while cercariae from 11 snails turned out to be S. bovis. Only two sites out of 40 namely Kisukioni and Katiwa, were active transmission sites. Both sites were active transmission sites for both S. haematobium and S. bovis. The assay reliably identified and distinguished between S. haematobium and S. bovis cercariae, even when only a few cercariae (5-10) were present in the sample, or when the parasite DNA concentrations were as low as five pico grammes (5pg). The FTA (R) paper offered a more reliable way of collecting, transporting and storing DNA material, and the samples. Conclusion: The PCR-based assay can potentially be used to support schistosomiasis control efforts, in epidemiological studies of urinary schistosomiasis, or in transmission ecology studies of S. haematobium and S. bovis.

Automated summary

The PCR-RFLP method targeting the ITS2 region of the ribosomal DNA was used to accurately identify mammalian schistosome cercariae shed by bulinid snails in freshwater habitats. This method can help determine transmission sites and assess the distribution of parasite species in the study area.