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      Prevalence of intestinal parasites in companion dogs with diarrhea in Beijing, China, and genetic characteristics of Giardia and Cryptosporidium species

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          Abstract

          Companion animals including dogs are one of the important components in One Health. Parasites may cause not only diseases in pet animals but also many zoonotic diseases infecting humans. In this study, we performed a survey of intestinal parasites in fecal specimens ( n = 485) collected from outpatient pet dogs with diarrhea in Beijing, China, for the entire year of 2015 by microscopic examination (all parasites) and SSU rRNA-based nested PCR detection ( Giardia and Cryptosporidium). We observed a total of 124 (25.6%) parasite-positive specimens that contained one or more parasites, including Giardia duodenalis (12.8%), Cryptosporidium spp. (4.9%), Cystoisospora spp. (4.3%), trichomonads (4.3%), Toxocara canis (3.5%), Trichuris vulpis (0.6%), and Dipylidium caninum (0.2%). Among the 55 dog breeds, infection rates were significantly higher in border collies and bulldogs, but lower in poodles ( p < 0.05). Risk factor analysis suggested that age was negatively correlated with the infection rate ( p < 0.00001), while vaccination and deworming in the past 12 months could significantly reduce the parasite infections ( p < 0.01). Among the 62 Giardia-positive specimens, 21 were successfully assigned into assemblages using glutamate dehydrogenase ( gdh) and/or beta-giardin ( bg) genes, including assemblage D ( n = 15), C ( n = 5), and F ( n = 1). Among the 24 Cryptosporidium-positive specimens by SSU rRNA PCR, 20 PCR amplicons could be sequenced and identified as Cryptosporidium canis ( n = 20). Collectively, this study indicates that parasites are a significant group of pathogens in companion dogs in Beijing, and companion dogs may potentially transmit certain zoonotic parasites to humans, particularly those with weak or weakened immunity.

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          Most cited references35

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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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            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
                gzhu@cvm.tamu.edu
                gaodifeihao@cau.edu.cn
                Journal
                Parasitol Res
                Parasitol. Res
                Parasitology Research
                Springer Berlin Heidelberg (Berlin/Heidelberg )
                0932-0113
                1432-1955
                18 November 2017
                2018
                : 117
                : 1
                : 35-43
                Affiliations
                [1 ]ISNI 0000 0004 0530 8290, GRID grid.22935.3f, College of Veterinary Medicine, , China Agricultural University, ; Haidian District, Beijing, 100193 China
                [2 ]ISNI 0000 0004 4687 2082, GRID grid.264756.4, Department of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, , Texas A&M University, ; College Station, TX 77843-4467 USA
                Author information
                http://orcid.org/0000-0003-3888-0659
                Article
                5631
                10.1007/s00436-017-5631-7
                7088013
                29150700
                4a9a530d-d7ad-498a-b68d-4d9a03ef1088
                © Springer-Verlag GmbH Germany 2017

                This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

                History
                : 6 August 2017
                : 20 September 2017
                Funding
                Funded by: National Innovation Experiment Program for Undergraduates of China
                Award ID: 201510019073
                Award Recipient :
                Funded by: Chinese Universities Scientific Fund for Postgraduates
                Award ID: 2015DY003
                Award Recipient :
                Categories
                Original Paper
                Custom metadata
                © Springer-Verlag GmbH Germany, part of Springer Nature 2018

                Parasitology
                companion dogs,beijing,giardia duodenalis,cryptosporidium canis,cystoisospora spp.,toxocara canis,trichomonads,risk factors

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