| Summary: | Enteric parasites including <i>Giardia duodenalis</i>, <i>Cryptosporidium</i> spp., and to a lesser extent, <i>Blastocystis</i> sp. and <i>Enterocytozoon bieneusi</i>, are major worldwide contributors to diarrhoeal disease. Assessing their molecular frequency and diversity is important to ascertain the sources of infection, transmission dynamics, and zoonotic potential. Little molecular information is available on the genotypes of these pathogens circulating in apparently healthy children. Here, we show that asymptomatic carriage of <i>G. duodenalis </i>(17.4%, 95% CI: 15.5‒19.4%), <i>Blastocystis</i> sp. (13.0%, 95% CI: 11.4‒14.8%), and <i>Cryptosporidium</i> spp. (0.9%, 95% CI: 0.5‒1.5%) is common in children (1‒16 years; <i>n</i> = 1512) from Madrid, Spain. Our genotyping data indicate that; (i) the observed frequency and diversity of parasite genetic variants are very similar to those previously identified in Spanish clinical samples, so that the genotype alone does not predict the clinical outcome of the infection, (ii) anthroponotic transmission accounts for a large proportion of the detected cases, highlighting that good personal hygiene practices are important to minimizing the risk of infection, (iii) <i>Blastocystis</i> ST4 may represent a subtype of the parasite with higher pathogenic potential, and (iv) <i>Enterocytozoon bieneusi</i> does not represent a public health concern in healthy children.
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