A New Method to Test Molluscicides against the Philippine Schistosomiasis Snail Vectors

Abstract

To expedite the discovery of novel molluscicides in the laboratory, this study aimed to evaluate the performance of a new molluscicidal assay. This assay is based on Oncomelania hupensis quadrasi snails and is called miniaturized plate test or mpt. To perform this assay, a 12-well plate, 3 snails per well, and 24-h exposure period were used. The performance of mpt was evaluated using niclosamide and Ardisia plant extract (tagpo extract) as test substances while WHO’s guidelines for a conventional plate test (cpt) served as standard. One cpt and four mpt independent trials were performed for niclosamide and tagpo extract. Probit analysis of dose–response data was run in R to generate lethal concentrations (LC50 and LC90), while lethal ratio test was performed to detect significant difference between paired LC50s (or LC90s). Using niclosamide, the calculated LC50 values were 0.104, 0.127, 0.136, 0.139, and 0.140 g/m2 for cpt, mpt 1, mpt 2, mpt 3, and mpt 4, respectively, while the LC90 values were 0.266, 0.268, 0.244, 0.251, and 0.261 g/m2, using the same sequence, respectively. For tagpo extract, the LC50 values were 1.467, 1.547, 1.659, 1.797, and 1.659 g/m2, for cpt, mpt 1, mpt 2, mpt 3, and mpt 4, respectively, and the LC90s were 2.188, 2.195, 2.501, 2.358, and 2.501 g/m2, respectively. The lethal ratio test revealed that a significant difference exists between the LC50s of cpt and mpt 1 when using niclosamide with a lethal ratio and confidence limits of 0.820 (0.663, 0.977, p<0.05) and another significant difference between LC50s of mpt 1 and mpt 3 using tagpo extract with computed lethal ratio and confidence limits of 0.861 (0.782, 0.939, p<0.05). Taken together, the results point out that mpt generates accurate and reproducible lethal concentration values. Hence, mpt may be used as an alternative method to screen molluscicides that are active against schistosome snail vectors.