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      Molecular and descriptive epidemiology of intestinal protozoan parasites of children and their pets in Cauca, Colombia: a cross-sectional study

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          Abstract

          Background

          Parasitic infections, particularly those caused by protozoa, represent a considerable public health problem in developing countries. Blastocystis, Giardia duodenalis, Cryptosporidium spp. and the Entamoeba complex ( Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii) are the most common etiological causes of intestinal parasitic infections.

          Methods

          We carried out a descriptive cross-sectional study in school-age children attending a daycare institution in commune eight of Popayán, Cauca (Southwest Colombia). A total of 266 fecal samples were collected (258 from children and eight from pets). Blastocystis, G. duodenalis, Cryptosporidium spp. and the Entamoeba complex were identified by microscopy, quantitative real-time PCR (qPCR) and conventional PCR. The concordance of qPCR and microscopy was assessed using the Kappa index. Molecular characterization was conducted to identify Blastocystis subtypes (18S), G. duodenalis assemblages ( tpi and gdh) and Cryptosporidium species/subtypes (18S and GP60). Potential associations between intestinal parasitism and sociodemographic factors were examined using bivariate analyses.

          Results

          A total of 258 fecal samples from children were analyzed by microscopy and 255 samples were analyzed by qPCR. The prevalence of Blastocystis was between 25.19% (microscopy) and 39.22% (qPCR), that of G. duodenalis was between 8.14% (microscopy) and 10.59% (qPCR), that of Cryptosporidium spp. was estimated at 9.8% (qPCR), and that of the Entamoeba complex was between 0.39% (conventional PCR) and 0.78% (microscopy). The concordance between microscopy and qPCR was very low. Blastocystis ST1 (alleles 4, 8, and 80), ST2 (alleles 11, 12, and 15), ST3 (alleles 31, 34, 36, 38,57, and 151), and ST4 (alleles 42 and 91), G. duodenalis assemblages AII, BIII, BIV and D, C. parvum subtype IIa and C. hominis subtype IbA9G3R2 were identified. The only identified member of the Entamoeba complex corresponded to E. histolytica. No statistically significant association was identified between parasitic infection and any sociodemographic variable.

          Conclusion

          This study revealed the usefulness of molecular methods to depict the transmission dynamics of parasitic protozoa in southwest Colombia. The presence of some of these protozoa in domestic animals may be involved in their transmission.

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          Most cited references35

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          New insights on classification, identification, and clinical relevance of Blastocystis spp.

          Blastocystis is an unusual enteric protozoan parasite of humans and many animals. It has a worldwide distribution and is often the most commonly isolated organism in parasitological surveys. The parasite has been described since the early 1900s, but only in the last decade or so have there been significant advances in our understanding of Blastocystis biology. However, the pleomorphic nature of the parasite and the lack of standardization in techniques have led to confusion and, in some cases, misinterpretation of data. This has hindered laboratory diagnosis and efforts to understand its mode of reproduction, life cycle, prevalence, and pathogenesis. Accumulating epidemiological, in vivo, and in vitro data strongly suggest that Blastocystis is a pathogen. Many genotypes exist in nature, and recent observations indicate that humans are, in reality, hosts to numerous zoonotic genotypes. Such genetic diversity has led to a suggestion that previously conflicting observations on the pathogenesis of Blastocystis are due to pathogenic and nonpathogenic genotypes. Recent epidemiological, animal infection, and in vitro host-Blastocystis interaction studies suggest that this may indeed be the case. This review focuses on such recent advances and also provides updates on laboratory and clinical aspects of Blastocystis spp.
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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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              Discrimination of all genotypes of Giardia duodenalis at the glutamate dehydrogenase locus using PCR-RFLP.

              A PCR-RFLP genotyping tool was developed and used to characterise morphologically identical isolates of Giardia duodenalis from a variety of host species. Primers were designed to amplify a 432bp region of the glutamate dehydrogenase gene (gdh) from genetic Assemblages AI, AII, BIII, BIV, C, D and E of G. duodenalis. DNA extracted from cultured Giardia trophozoites, Giardia cysts purified from faeces and directly from whole faeces was amplified and sequenced at the gdh and 18SrDNA loci. The gdh sequences were identical with published gdh sequences for each assemblage with a few exceptions. However, in some cases genotyping results obtained using gdh differed from 18SrDNA genotyping results. From gdh sequence information a PCR-RFLP profile was identified for each of the genetic assemblages. PCR-RFLP is a reproducible, reliable and sensitive method for genotyping Giardia. Eight human, 12 cat, 9 dog and 16 cattle faecal isolates were genotyped using PCR-RFLP. This method allows G. duodenalis isolates from human-beings, their companion animals and livestock to be genotyped directly from faeces, leading to valuable information about Giardia genotypes in population without the need for in vitro/in vivo amplification.
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                Author and article information

                Contributors
                ximenasoler31@gmail.com
                adrianahiguera81@gmail.com
                gio9024@gmail.com
                lreinel@unicauca.edu.co
                lbuitron@unicauca.edu.co
                lmunoz@unicauca.edu.co
                fgonzal@unicauca.edu.co
                mclopez@gmail.com
                jcesargiraldo@gmail.com
                juand.ramirez@urosario.edu.co
                Journal
                BMC Infect Dis
                BMC Infect. Dis
                BMC Infectious Diseases
                BioMed Central (London )
                1471-2334
                26 February 2019
                26 February 2019
                2019
                : 19
                : 190
                Affiliations
                [1 ]ISNI 0000 0001 2205 5940, GRID grid.412191.e, Grupo de Investigaciones Microbiológicas-UR (GIMUR), Programa de Biología, Facultad de Ciencias Naturales y Matemáticas, , Universidad del Rosario, ; Bogotá, Colombia
                [2 ]ISNI 0000 0001 2158 6862, GRID grid.412186.8, Centro de estudios en Microbiología y Parasitología (CEMPA), Departamento de Medicina Interna, Facultad de Ciencias de la Salud, , Universidad del Cauca, ; Popayán, Colombia
                [3 ]ISNI 0000 0001 0286 3748, GRID grid.10689.36, Departamento de Salud Pública, Facultad de Medicina, , Universidad Nacional de Colombia, ; Bogotá, Colombia
                [4 ]GRID grid.442170.4, Programa de Biología, , Universidad INCCA de Colombia, ; Bogotá, Colombia
                Author information
                http://orcid.org/0000-0002-1344-9312
                Article
                3810
                10.1186/s12879-019-3810-0
                6390308
                30808303
                82b00bc5-2be1-4f45-8aa9-56f11262fd63
                © The Author(s). 2019

                Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 28 November 2018
                : 12 February 2019
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100007637, Departamento Administrativo de Ciencia, Tecnología e Innovación;
                Award ID: 122271250521
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2019

                Infectious disease & Microbiology
                blastocystis,giardia duodenalis,cryptosporidium,entamoeba histolytica/dispar/moshkovskii complex,zoonotic disease

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