Identification of Zoonotic Genotypes of Giardia duodenalis

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Abstract
Giardia duodenalis, originally regarded as a commensal organism, is the etiologic agent of giardiasis, a gastrointestinal disease of humans and animals. Giardiasis causes major public and veterinary health concerns worldwide. Transmission is either direct, through the faecal-oral route, or indirect, through ingestion of contaminated water or food. Genetic characterization of G. duodenalis isolates has revealed the existence of seven groups (assemblages A to G) which differ in their host distribution. Assemblages A and B are found in humans and in many other mammals, but the role of animals in the epidemiology of human infection is still unclear, despite the fact that the zoonotic potential of Giardia was recognised by the WHO some 30 years ago. Here, we performed an extensive genetic characterization of 978 human and 1440 animal isolates, which together comprise 3886 sequences from 4 genetic loci. The data were assembled into a molecular epidemiological database developed by a European network of public and veterinary health Institutions. Genotyping was performed at different levels of resolution (single and multiple loci on the same dataset). The zoonotic potential of both assemblages A and B is evident when studied at the level of assemblages, sub-assemblages, and even at each single locus. However, when genotypes are defined using a multi-locus sequence typing scheme, only 2 multi-locus genotypes (MLG) of assemblage A and none of assemblage B appear to have a zoonotic potential. Surprisingly, mixtures of genotypes in individual isolates were repeatedly observed. Possible explanations are the uptake of genetically different Giardia cysts by a host, or subsequent infection of an already infected host, likely without overt symptoms, with a different Giardia species, which may cause disease. Other explanations for mixed genotypes, particularly for assemblage B, are substantial allelic sequence heterogeneity and/or genetic recombination. Although the zoonotic potential of G. duodenalis is evident, evidence on the contribution and frequency is (still) lacking. This newly developed molecular database has the potential to tackle intricate epidemiological questions concerning protozoan diseases. Giardia duodenalis is a parasite causing a gastrointestinal disease in humans, pets, livestock, and wildlife. The role of animals in human disease is unclear, because Giardia from humans and animals is morphologically indistinguishable. An international consortium of both veterinary and public health institutions built a web-based database, where molecular and epidemiological data are combined. After extensive genetic characterization, the zoonotic potential of Giardia became evident, but data on frequency and role in epidemiology is (still) lacking. Surprisingly, mixtures of Giardia genotypes in individual hosts were frequently observed, and have important implications for the etiology of Giardiasis. Possible explanations are the uptake of mixtures of Giardia genotypes by one host, or subsequent infection of an already infected host, likely without overt symptoms, with a different Giardia species, which may cause disease. We demonstrated that collaborative, human and veterinary health integrated databases have the potential to tackle intricate epidemiological questions concerning parasitic diseases, as was demonstrated for G. duodenalis in the present study.