Polymerase chain reaction and restriction fragment length polymorphism analysis of the ITS2 region for differatiation of Brazilian Biomphalaria intermediate hosts of the Schistosoma mansoni

Vidigal, Teofânia H. D. A.; Magalhães, Kelly Grace; Carvalho, Omar dos Santos

doi:10.1590/s0037-86822004000400012

Cited by 6 publications

(4 citation statements)

References 12 publications

(10 reference statements)

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“…Once in the laboratory, snails collected were sorted (based both on their shell morphology [15] and polymerase chain reaction restriction fragment length polymorphism analysis (PCR-RFLP) of the ITS2 region [16]) and distributed individually in 100 ml well labeled plastic boxes containing spring water. A piece of polystyrene was added in each rearing box for eggs laying.…”

Section: Methodsmentioning

confidence: 99%

Biomphalaria camerunensis as a viable alternative intermediate host for Schistosoma mansoni in southern Cameroon

et al. 2018

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BackgroundIntestinal schistosomiasis due to Schistosoma mansoni was mapped in Cameroon in the 1990s and preventive chemotherapy launched since 2005. A situation analysis conducted in 2011 revealed an increase in schistosomiasis transmission, especially in the equatorial part of the country, despite the fact that Biomphalaria pfeifferi, the main intermediate host of this parasite, is now scarce in many foci. Biomphalaria camerunensis, restricted to the equatorial part of the country, is considered as a less suitable host for S. mansoni due to it resistance to the parasite, although exhibiting a better survival than B. pfeifferi. In a context where human migration is quite frequent as a consequence of terrorism, war in neighboring countries, as well as development of hydraulic projects, it seems appropriate to evaluate the current epidemiological role of B. camerunensis to estimate the risk of extension of S. mansoni distribution in Cameroon. To do this, the susceptibility of three B. pfeifferi and five B. camerunensis populations to a strain of S. mansoni was assessed. Juvenile snails (G1) of each population were infected with S. mansoni miracidia, and prepatent period, infection and survival rates of infected snails, as well as cercarial production were recorded and compared between snail species and populations.ResultsCompatibility tests were performed on a total of 827 snails: 344 B. pfeifferi and 483 B. camerunensis. Infection rates were quite heterogeneous, higher in B. pfeifferi (61.5%) as compared to B. camerunensis (7.8%) (Chi-square test: χ2 = 258.88, df = 1, P < 0.0001). All the three B. pfeifferi-infected populations were susceptible to S. mansoni. However, among the five B. camerunensis populations tested, four were susceptible to S. mansoni, with 21.9% as the highest infection rate. The prepatent period was, on average, shorter in B. pfeifferi than in B. camerunensis (P < 0.0001), but the cercarial production was significantly higher in B. camerunensis as compared to B. pfeifferi (P < 0.001).ConclusionThese findings indicate that B. camerunensis populations might substantially contribute to S. mansoni transmission and dissemination in Cameroon, their low susceptibility being compensated by their high cercariae production. More attention and surveillance towards this species are required to achieve intestinal schistosomiasis elimination in Cameroon.Electronic supplementary materialThe online version of this article (10.1186/s13071-018-2763-2) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Biomphalaria camerunensis as a viable alternative intermediate host for Schistosoma mansoni in southern Cameroon

et al. 2018

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“…PCR amplification and RFLP conditions, using HpaII enzyme were the same as used by Vidigal et al (2004). Products were visualised on 6% silver stained polyacrylamide gels and the results were recorded with the camera Mavica (Sony).…”

Section: Methodsmentioning

confidence: 99%

Diagnostic of Biomphalaria snails and Schistosoma mansoni: DNA obtained from traces of shell organic materials

Caldeira

Jannotti-Passos

Lira

et al. 2004

Mem. Inst. Oswaldo Cruz

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“…In the last two decades, molecular tools have been evaluated for both snail identification and phylogenetic studies [26][27][28][29]; the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis of the internal transcribed spacer (ITS) region of rDNA and the analysis of cytochrome c oxidase subunit 1 region of mitochondrial DNA have proven to be cheaper and powerful for the identification of several Biomphalaria species [30][31][32][33][34][35]. More recently, the PCR-RFLP technique has been used to distinguish Biomphalaria species in Brazil and it appears to be an alternative molecular tool to their morphological identification [36].…”

Section: Introductionmentioning

confidence: 99%

“…The aim of this study was to test the efficiency of the PCR-RFLP protocol developed for identification of Biomphalaria species in South America [34,36], for the separation of the two main Biomphalaria spp. currently found in Cameroon.…”

Section: Introductionmentioning

confidence: 99%

PCR-based molecular identification of two intermediate snail hosts of Schistosoma mansoni in Cameroon

et al. 2020

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Background Snails of the genus Biomphalaria are intermediate hosts of Schistosoma mansoni, the causative agent of the human intestinal schistosomiasis. Two Biomphalaria species (Biomphalaria pfeifferi and Biomphalaria camerunensis) are involved in the transmission in Cameroon, where the disease is present nationwide. However, difficulty in the identification of both vectors impedes proper assessment of the epidemiological burden caused by each species. To overcome this issue, we designed a PCR-based molecular diagnostic tool to improve the identification of these species. Methods We analyzed the internal transcribed spacer 2 (ITS2) region of Biomphalaria ribosomal DNA (rDNA) using polymerase chain reaction amplification (PCR) and restriction fragment length polymorphism (RFLP). Results The amplification of the ITS2 region of Biomphalaria snails resulted in a 490 bp fragment and produced two profiles for each species after digestion with the restriction enzyme Hpa II. The profile 1 (Bc-HpaII-1: 212-bp and 139-bp bands) for B. camerunensis, was common in all the sampling points; the profile 2 (Bc-HpaII-2: 212-bp and 189-bp bands), was only observed in the Lake Monoun Njindoun sampling site. Biomphalaria pfeifferi profile 1 (Bpf-HpaII-1: 211-bp and 128-bp bands) was common in most of B. pfeifferi sampling points; the profile 2 (Bpf-HpaII-2: 289-bp and 128-bp bands) was only observed in Mokolo (Far North Cameroon).The second restriction enzyme TaqαI, revealed three band profiles, Bc-TaqαI-1 (243-bp, 136-bp and 118-bp bands) and Bc-TaqαI-2 (244-bp, 136-bp and 99-bp) for B. camerunensis and Bpf-TaqαI-1 (242-bp, 135-bp and 107-bp bands) for B. pfeifferi. Sequencing analysis revealed the occurrence of six haplotypes for B. camerunensis and three haplotypes for B. pfeifferi. The level of gene flow was low and the Biomphalaria populations were not in demographic expansion according to neutrality tests (Tajima’s D and Fu’s Fs). Conclusions The PCR-RFLP technique revealed genetic diversity in Biomphalaria snails, and the combination with the morphological method could improve the identification of B. pfeifferi and B. camerunensis in Cameroon. This could help focus on the infection to evaluate the transmission risk with respect of the different species and to develop efficient and cost-effective control measures.

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Polymerase chain reaction and restriction fragment length polymorphism analysis of the ITS2 region for differatiation of Brazilian Biomphalaria intermediate hosts of the Schistosoma mansoni

Cited by 6 publications

References 12 publications

Biomphalaria camerunensis as a viable alternative intermediate host for Schistosoma mansoni in southern Cameroon

Biomphalaria camerunensis as a viable alternative intermediate host for Schistosoma mansoni in southern Cameroon

Diagnostic of Biomphalaria snails and Schistosoma mansoni: DNA obtained from traces of shell organic materials

PCR-based molecular identification of two intermediate snail hosts of Schistosoma mansoni in Cameroon

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