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      Molecular characterisation of protist parasites in human-habituated mountain gorillas ( Gorilla beringei beringei), humans and livestock, from Bwindi impenetrable National Park, Uganda

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          Abstract

          Background

          Over 60 % of human emerging infectious diseases are zoonotic, and there is growing evidence of the zooanthroponotic transmission of diseases from humans to livestock and wildlife species, with major implications for public health, economics, and conservation. Zooanthroponoses are of relevance to critically endangered species; amongst these is the mountain gorilla ( Gorilla beringei beringei) of Uganda. Here, we assess the occurrence of Cryptosporidium, Cyclospora, Giardia, and Entamoeba infecting mountain gorillas in the Bwindi Impenetrable National Park (BINP), Uganda, using molecular methods. We also assess the occurrence of these parasites in humans and livestock species living in overlapping/adjacent geographical regions.

          Results

          Diagnostic PCR detected Cryptosporidium parvum in one sample from a mountain gorilla (IIdA23G2) and one from a goat (based on SSU). Cryptosporidium was not detected in humans or cattle. Cyclospora was not detected in any of the samples analysed. Giardia was identified in three human and two cattle samples, which were linked to assemblage A, B and E of G. duodenalis. Sequences defined as belonging to the genus Entamoeba were identified in all host groups. Of the 86 sequence types characterised, one, seven and two have been recorded previously to represent genotypes of Cryptosporidium, Giardia, and Entamoeba, respectively, from humans, other mammals, and water sources globally.

          Conclusions

          This study provides a snapshot of the occurrence and genetic make-up of selected protists in mammals in and around BINP. The genetic analyses indicated that 54.6% of the 203 samples analysed contained parasites that matched species, genotypes, or genetic assemblages found globally. Seventy-six new sequence records were identified here for the first time. As nothing is known about the zoonotic/zooanthroponotic potential of the corresponding parasites, future work should focus on wider epidemiological investigations together with continued surveillance of all parasites in humans, other mammals, the environment, and water in this highly impoverished area.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s13071-017-2283-5) contains supplementary material, which is available to authorized users.

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          Most cited references85

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          Triosephosphate Isomerase Gene Characterization and Potential Zoonotic Transmission of Giardia duodenalis

          To address the source of infection in humans and public health importance of Giardia duodenalis parasites from animals, nucleotide sequences of the triosephosphate isomerase (TPI) gene were generated for 37 human isolates, 15 dog isolates, 8 muskrat isolates, 7 isolates each from cattle and beavers, and 1 isolate each from a rat and a rabbit. Distinct genotypes were found in humans, cattle, beavers, dogs, muskrats, and rats. TPI and small subunit ribosomal RNA (SSU rRNA) gene sequences of G. microti from muskrats were also generated and analyzed. Phylogenetic analysis on the TPI sequences confirmed the formation of distinct groups. Nevertheless, a major group (assemblage B) contained most of the human and muskrat isolates, all beaver isolates, and the rabbit isolate. These data confirm that G. duodenalis from certain animals can potentially infect humans and should be useful in the detection, differentiation, and taxonomy of Giardia spp.
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            Genetic heterogeneity at the beta-giardin locus among human and animal isolates of Giardiaduodenalis and identification of potentially zoonotic subgenotypes.

            Human giardiasis, caused by the intestinal flagellate Giardia duodenalis, is considered a zoonotic infection, although the role of animals in the transmission to humans is still unclear. Molecular characterisation of cysts of human and animal origin represents an objective means to validate or reject this hypothesis. In the present work, cysts were collected in Italy from humans (n=37) and animals (dogs, one cat and calves, n=46), and were characterised by PCR amplification and sequencing of the beta-giardin gene. As expected, only Assemblages A and B were identified among human isolates. The host-specific Assemblages C and D were found in the majority of dog isolates; however, 6 dog isolates were typed as Assemblage A. The cat-specific Assemblage F has been identified in the single feline isolate available. Among calf isolates, most were typed as Assemblages A (n=12) and B (n=5), whereas the host-specific Assemblage E was rarely found (n=3). Sequence heterogeneity in the beta-giardin gene allowed a number of subgenotypes to be identified within Assemblage A (8 subgenotypes), B (6 subgenotypes), D (2 subgenotypes), and E (3 subgenotypes). Five of these subgenotypes, namely A1, A2, A3, A4 and B3, were found to be associated with infections of humans, of dogs and of calves; these data, therefore, supported the role of these animals as a source of infection for humans.
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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
                mnolan@rvc.ac.uk
                munger4@rvc.ac.uk
                yyeap3@rvc.ac.uk
                erogers3@rvc.ac.uk
                ilary.millet@edu.unito.it
                kharman@rvc.ac.uk
                mfox@rvc.ac.uk
                Gladys@ctph.org
                dblake@rvc.ac.uk
                Journal
                Parasit Vectors
                Parasit Vectors
                Parasites & Vectors
                BioMed Central (London )
                1756-3305
                18 July 2017
                18 July 2017
                2017
                : 10
                : 340
                Affiliations
                [1 ]ISNI 0000 0001 2161 2573, GRID grid.4464.2, Department of Pathobiology and Population Sciences, Royal Veterinary College, , University of London, ; Hatfield, UK
                [2 ]Conservation through Public Health, Plot 3 Mapera Lane, Uringi Crescent, Entebbe, Uganda
                Article
                2283
                10.1186/s13071-017-2283-5
                5516388
                6697801d-805c-4af7-b6d9-a714ccad3c7c
                © The Author(s). 2017

                Open AccessThis 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
                : 14 March 2017
                : 11 July 2017
                Categories
                Research
                Custom metadata
                © The Author(s) 2017

                Parasitology
                zoonosis,zooanthroponosis,infectious disease,cryptosporidium,giardia,entamoeba
                Parasitology
                zoonosis, zooanthroponosis, infectious disease, cryptosporidium, giardia, entamoeba

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