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Genotyping of Giardia in Dutch patients and animals: a phylogenetic analysis of human and animal isolates
Authors:van der Giessen J W B  de Vries A  Roos M  Wielinga Peter  Kortbeek L M  Mank T G
Institution:a National Institute for Public Health and the Environment (RIVM), Microbiological Laboratory for Health Protection, Antonie van Leeuwenhoeklaan 9, P.O. Box 1, Bilthoven, The Netherlands
b National Institute of Public Health and the Environment (RIVM), Laboratory for Infectious Diseases Surveillance and Screening, Antonie van Leeuwenhoeklaan 9, P.O. Box 1, Bilthoven, The Netherlands
c Laboratory of Public Health, Department of Parasitology, Boerhaavelaan 26, 2035 RC, Haarlem, The Netherlands
Abstract:Giardia duodenalis (syn. Giardia lamblia, Giardia intestinalis) is a protozoan organism that can infect the intestinal tract of many animal species including mammals. Genetic heterogeneity of G. duodenalis is well described but the zoonotic potential is still not clear. In this study, we analysed 100 Giardia DNA samples directly isolated from human stool specimens, to get more insight in the different G. duodenalis assemblages present in the Dutch human population. Results showed that these human isolates could be divided into two main Assemblages A and B within the G. duodenalis group on the basis of PCR assays specific for the Assemblages A and B and the DNA sequences of 18S ribosomal RNA and the glutamate dehydrogenase (gdh) genes. Genotyping results showed that G. duodenalis isolates originating from Dutch human patients belonged in 35% of the cases to Assemblage A (34/98) and in 65% of the cases to Assemblage B (64/98) whereas two human cases remained negative in all assays tested. In addition, we compared these human samples with animal samples from the Netherlands and human and animal samples from other countries. A phylogenetic analysis was carried out on the DNA sequences obtained from these Giardia and those available in GenBank. Using gdh DNA sequence analysis, human and animal Assemblage A and B Giardia isolates could be identified. However, phylogenetic analysis revealed different sub-clustering for human and animal isolates where host-species-specific assemblages (C, D, E, F and G) could be identified. The geographic origin of the human and animal samples was not a discriminating factor.
Keywords:Giardia  Molecular genetics  Phylogenetic analysis  18S rRNA  Glutamate dehydrogenase  Zoonotic disease
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