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      First description of Giardia duodenalis in buffalo calves ( Bubalus bubalis) in southwest region of São Paulo State, Brazil

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          Abstract

          We performed molecular characterization of Giardia duodenalis in buffalo calves from the Southwest region of São Paulo State, Brazil. A total of 183 fecal samples of Murrah breed buffaloes up to six months of age were collected. We examined these samples by the polymerase chain reaction (PCR) targeting the small-subunit ribosomal RNA gene and positive samples were characterized using additional PCR assays targeting a portion of the beta-giardin, the glutamate dehydrogenase and the triose-phosphate isomerase genes. Based on the SSU rRNA nPCR, the presence of G. duodenalis was confirmed in 12 (6.56%) of fecal samples, of these, five, four and three samples were positive for the tpi, bg and gdh genes, respectively. Assemblage identification by sequencing was successful in 6 of 12 samples and sequence analysis showed 100% genetic similarity with G. duodenalis assemblage E. This observation represents the first detection of G. duodenalis assemblage E in buffaloes calves in Brazil.

          Highlights

          • First study of Giardia in fecal samples of buffalo calves from Brazil.

          • Detection of Giardia in buffaloes by nested PCR using four genetic markers.

          • Phylogenetic analysis identified Giardia duodenalis assemblage E.

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          Most cited references27

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          Zoonotic potential and molecular epidemiology of Giardia species and giardiasis.

          Molecular diagnostic tools have been used recently in assessing the taxonomy, zoonotic potential, and transmission of Giardia species and giardiasis in humans and animals. The results of these studies have firmly established giardiasis as a zoonotic disease, although host adaptation at the genotype and subtype levels has reduced the likelihood of zoonotic transmission. These studies have also identified variations in the distribution of Giardia duodenalis genotypes among geographic areas and between domestic and wild ruminants and differences in clinical manifestations and outbreak potentials of assemblages A and B. Nevertheless, our efforts in characterizing the molecular epidemiology of giardiasis and the roles of various animals in the transmission of human giardiasis are compromised by the lack of case-control and longitudinal cohort studies and the sampling and testing of humans and animals living in the same community, the frequent occurrence of infections with mixed genotypes and subtypes, and the apparent heterozygosity at some genetic loci for some G. duodenalis genotypes. With the increased usage of multilocus genotyping tools, the development of next-generation subtyping tools, the integration of molecular analysis in epidemiological studies, and an improved understanding of the population genetics of G. duodenalis in humans and animals, we should soon have a better appreciation of the molecular epidemiology of giardiasis, the disease burden of zoonotic transmission, the taxonomy status and virulences of various G. duodenalis genotypes, and the ecology of environmental contamination.
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            Zoonotic potential of Giardia.

            Giardia duodenalis (syn. Giardia lamblia and Giardia intestinalis) is a common intestinal parasite of humans and mammals worldwide. Assessing the zoonotic transmission of the infection requires molecular characterization as there is considerable genetic variation within G. duodenalis. To date eight major genetic groups (assemblages) have been identified, two of which (A and B) are found in both humans and animals, whereas the remaining six (C to H) are host-specific and do not infect humans. Sequence-based surveys of single loci have identified a number of genetic variants (genotypes) within assemblages A and B in animal species, some of which may have zoonotic potential. Multi-locus typing data, however, has shown that in most cases, animals do not share identical multi-locus types with humans. Furthermore, interpretation of genotyping data is complicated by the presence of multiple alleles that generate "double peaks" in sequencing files from PCR products, and by the potential exchange of genetic material among isolates, which may account for the non-concordance in the assignment of isolates to specific assemblages. Therefore, a better understanding of the genetics of this parasite is required to allow the design of more sensitive and variable subtyping tools, that in turn may help unravel the complex epidemiology of this infection. Copyright © 2013. Published by Elsevier Ltd.
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              Molecular characterization of human isolates of Giardia duodenalis from Ethiopia.

              Giardia duodenalis, a flagellated protozoan, represents a common cause of gastroenteritis in Ethiopia, however very little information is available on the epidemiology and transmission routes of this pathogen, and a genetic characterization of the parasite has never been attempted in this country. The aim of this study was the genetic analysis of human isolates of G. duodenalis collected in different localities across the country, both from urban and rural areas. A fragment of the beta-giardin gene was amplified by nested PCR and analyzed by restriction and sequence analyses. Of the 59 isolates examined, 31 (52%) were typed as assemblage A and 13 (22%) as assemblage B. A strong correlation between the presence of symptoms and infection with assemblage B was observed. The remaining 15 (25%) isolates were typed as mixed infections by PCR-RFLP, specifically, A+F (in seven isolates) and A+B (in eight isolates). Sequencing of the A+F products confirmed the presence of assemblage F in three isolates, whereas the remaining four were identified as assemblage A. The detection of assemblage F, a cat-specific assemblage that to date has not been associated with human infections, was not able to be confirmed by the analysis of two commonly used markers (small subunit ribosomal RNA and triosephosphate isomerase). The analysis of the one isolate that was successfully amplified with the glutamate dehydrogenase primers unambiguously identified it as G. duodenalis, yet it was distinct from the established A and F sequences; thus the exact genetic identity of these isolates remains unclear.
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                Author and article information

                Contributors
                Journal
                Food Waterborne Parasitol
                Food Waterborne Parasitol
                Food and Waterborne Parasitology
                Elsevier
                2405-6766
                31 May 2019
                September 2019
                31 May 2019
                : 16
                : e00062
                Affiliations
                [a ]Universidade Estadual Paulista (Unesp), Faculdade de Medicina Veterinária, Araçatuba, São Paulo, Brazil
                [b ]Universidade Estadual de Santa Cruz (UESC), Ilhéus, Bahia, Brazil
                [c ]Department of Infectious Disease & Global Health, Cummings School of Veterinary Medicine at Tufts University, North Grafton, Massachusetts, USA
                Author notes
                [* ]Corresponding author at: Universidade Estadual Paulista (Unesp), Faculdade de Medicina Veterinária, Araçatuba. Rua Clóvis Pestana, 793, Jardim Dona Amélia, cep 16050-680, Araçatuba, São Paulo, Brasil. katia.bresciani@ 123456unesp.br
                Article
                S2405-6766(19)30003-4 e00062
                10.1016/j.fawpar.2019.e00062
                7034009
                4dd621ab-df3f-44d2-b1ea-cba0c25f7a60
                © 2019 Published by Elsevier Inc. on behalf of International Association of Food and Waterborne Parasitology.

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 16 November 2018
                : 5 May 2019
                : 27 May 2019
                Categories
                Article

                buffaloes,genotype,giardiasis,multilocus genotyping
                buffaloes, genotype, giardiasis, multilocus genotyping

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