Abstract
Giardiasis in humans is a gastrointestinal disease transmitted by the potentially zoonotic Giardia duodenalis genotypes (assemblages) A and B. Small wild rodents such as mice and voles are discussed as potential reservoirs for G. duodenalis but are predominantly populated by the two rodent species Giardia microti and Giardia muris. Currently, the detection of zoonotic and non-zoonotic Giardia species and genotypes in these animals relies on cumbersome PCR and sequencing approaches of genetic marker genes. This hampers the risk assessment of potential zoonotic Giardia transmissions by these animals. Here, we provide a workflow based on newly developed real-time PCR schemes targeting the small ribosomal RNA multi-copy gene locus to distinguish G. muris, G. microti and G. duodenalis infections. For the identification of potentially zoonotic G. duodenalis assemblage types A and B, an established protocol targeting the single-copy gene 4E1-HP was used. The assays were specific for the distinct Giardia species or genotypes and revealed an analytical sensitivity of approximately one or below genome equivalent for the multi-copy gene and of about 10 genome equivalents for the single-copy gene. Retesting a biobank of small rodent samples confirmed the specificity. It further identified the underlying Giardia species in four out of 11 samples that could not be typed before by PCR and sequencing. The newly developed workflow has the potential to facilitate the detection of potentially zoonotic and non-zoonotic Giardia species in wild rodents.
Highlights
Giardia spp. are flagellated protozoan parasites of vertebrates and a frequent cause of gastrointestinal disease in wild and domestic animals and humans [1]
B are found in humans and a wide range of other mammals, including rodents, and are considered potentially zoonotic [2]; assemblages C/D are found in canids; assemblage E
Small rodents can be infected with the distinct species Giardia muris and Giardia microti [1,3,4]
Summary
Giardia spp. are flagellated protozoan parasites of vertebrates and a frequent cause of gastrointestinal disease in wild and domestic animals and humans [1]. Small rodents can be infected with the distinct species Giardia muris and Giardia microti [1,3,4] For both G. microti and, in particular, for. The zoonotic G. duodenalis assemblages A and B are genetically very distinct, and each group can be further subclassified based on genetic information determined by multi-locus sequence typing (MLST) [6]. Within these subgroups, assemblage AI forms a pan-global, almost clonal subpopulation that is found in a wide range of vertebrates, including humans, whereas assemblage AII seems very genetically diverse and is almost exclusively found in humans [1]. The subclassification of assemblage B is less well-defined—in particular, when
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