Non-parametric estimators of species richness are some of the extrapolation methods used in the study of biodiversity. However, communities or assemblages of parasites have received little attention. Therefore, in the present study, real data on the seasonal diversity of digeneans in the first intermediate host Heleobia parchappii at Nahuel Rucá shallow lake (Argentina) were used to evaluate seven non-parametric estimators of species richness, analyzing their behavior and their ability to estimate richness using different subsample sizes. In addition, the minimum sample size necessary to include all observed species was estimated. The Chao 1, Chao 2 and Bootstrap estimators were generally the best-performing estimators considering the evaluation criteria used. Given the significant role of larval digenean assemblages in the first intermediate host for nonparametric estimator calculations, it is suggested to employ Chao 1 and 2 when the proportion of rare species exceeds 30%. However, if this proportion is less than 30%, the use of Bootstrap is recommended. Furthermore, to obtain a more precise measure of biodiversity in these systems, it is suggested to increase the traditionally examined host number from 300 to 400 individuals.