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      Evaluation of prevalence and risk factors associated with Cryptosporidium infection in rural population of district Buner, Pakistan

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          Abstract

          Background

          Cryptosporidium spp are important intestinal protozoan parasites that cause diarrhea in humans, domestic and wild animals. Its infection remains a main public health concern however, the epidemics in human being is still unclear, particularly in developing countries. There are several factors that may enhance the spreading of this parasite in human population especially in young children.

          Methodology

          A questionnaire was designed to obtain the demographic and clinical data from the participants. A total of 425 stool samples were collected from suspected children (aged 3–10 years) in different hospitals and villages. The initial screening was performed with modified Ziehl Neelsen (mZN) staining technique followed by polymerase chain reaction (PCR). Several potential risk factors were also assessed through the obtained information from suspected individuals.

          Results

          Out of all 425 collected samples, 127 were observed positive by mZN with a prevalence of 29.88% (127/425). The 127 mZN positive samples together with 50 mZN negative samples were processed for molecular analysis through PCR assay. Among them, 71 out of 127 mZN positive samples and 4 out of 50 mZN negative samples were found positive by PCR. The molecular analysis showed that Cryptosporidium parvum was the main cause of infection in children. The results revealed that individuals exposed to diarrhea were more likely to be infected with Cryptosporidium infection while several environmental factors may also play a key role in spreading of this parasite.

          Conclusions/Significance of the study

          The current high prevalence of Cryptosporidium infection may be due to the lack of awareness and routine based testing in identification of this parasite in District Buner. Further studies are required to determine the importance of Cryptosporidium infection in this area as well as across the country and to find out the possible risk factors that may be associated with the occurrence of this protozoan. There is, however, an urgent need for laboratory-based observational studies to develop a more dynamic estimate of the cryptosporidial disease burden in the region.

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          Most cited references56

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          Cryptosporidium species in humans and animals: current understanding and research needs.

          Cryptosporidium is increasingly recognized as one of the major causes of moderate to severe diarrhoea in developing countries. With treatment options limited, control relies on knowledge of the biology and transmission of the members of the genus responsible for disease. Currently, 26 species are recognized as valid on the basis of morphological, biological and molecular data. Of the nearly 20 Cryptosporidium species and genotypes that have been reported in humans, Cryptosporidium hominis and Cryptosporidium parvum are responsible for the majority of infections. Livestock, particularly cattle, are one of the most important reservoirs of zoonotic infections. Domesticated and wild animals can each be infected with several Cryptosporidium species or genotypes that have only a narrow host range and therefore have no major public health significance. Recent advances in next-generation sequencing techniques will significantly improve our understanding of the taxonomy and transmission of Cryptosporidium species, and the investigation of outbreaks and monitoring of emerging and virulent subtypes. Important research gaps remain including a lack of subtyping tools for many Cryptosporidium species of public and veterinary health importance, and poor understanding of the genetic determinants of host specificity of Cryptosporidium species and impact of climate change on the transmission of Cryptosporidium.
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            Waterborne transmission of protozoan parasites: review of worldwide outbreaks - an update 2004-2010.

            The present update gives a comprehensive review of worldwide waterborne parasitic protozoan outbreaks that occurred and were published globally between January 2004 and December 2010. At least one hundred and ninety-nine outbreaks of human diseases due to the waterborne transmission of parasitic protozoa occurred and were reported during the time period from 2004 to 2010. 46.7% of the documented outbreaks occurred on the Australian continent, 30.6% in North America and 16.5% in Europe. Cryptosporidium spp. was the etiological agent in 60.3% (120) of the outbreaks, Giardia lamblia in 35.2% (70) and other protozoa in 4.5% (9). Four outbreaks (2%) were caused by Toxoplasma gondii, three (1.5%) by Cyclospora cayetanensis. In two outbreaks (1%) Acanthamoeba spp. was identified as the causative agent. In one outbreak, G. lamblia (in 17.6% of stool samples) and Cryptosporidium parvum (in 2.7% of stool samples) as well as Entamoeba histolytica (in 9.4% of stool samples) and Blastocystis hominis (in 8.1% of stool samples) were detected. In those countries that are likely affected most a lack of surveillance systems is noticeable. However, countries that established surveillance systems did not establish an international standardization of reporting systems. Copyright © 2011 Elsevier Ltd. All rights reserved.
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              Cryptosporidium pathogenicity and virulence.

              Cryptosporidium is a protozoan parasite of medical and veterinary importance that causes gastroenteritis in a variety of vertebrate hosts. Several studies have reported different degrees of pathogenicity and virulence among Cryptosporidium species and isolates of the same species as well as evidence of variation in host susceptibility to infection. The identification and validation of Cryptosporidium virulence factors have been hindered by the renowned difficulties pertaining to the in vitro culture and genetic manipulation of this parasite. Nevertheless, substantial progress has been made in identifying putative virulence factors for Cryptosporidium. This progress has been accelerated since the publication of the Cryptosporidium parvum and C. hominis genomes, with the characterization of over 25 putative virulence factors identified by using a variety of immunological and molecular techniques and which are proposed to be involved in aspects of host-pathogen interactions from adhesion and locomotion to invasion and proliferation. Progress has also been made in the contribution of host factors that are associated with variations in both the severity and risk of infection. Here we provide a review comprised of the current state of knowledge on Cryptosporidium infectivity, pathogenesis, and transmissibility in light of our contemporary understanding of microbial virulence.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ResourcesRole: SoftwareRole: VisualizationRole: Writing – original draft
                Role: ConceptualizationRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: Writing – review & editing
                Role: Data curationRole: ResourcesRole: Writing – original draft
                Role: ConceptualizationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ResourcesRole: Writing – original draft
                Role: SupervisionRole: ValidationRole: Writing – original draft
                Role: ConceptualizationRole: Formal analysisRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                2 January 2019
                2019
                : 14
                : 1
                : e0209188
                Affiliations
                [1 ] Department of Zoology, Abdul Wali Khan University Mardan, Mardan, Pakistan
                [2 ] Centre for Biotechnology and Microbiology University of Swat, Swat, Pakistan
                [3 ] Department of Environmental Science, University of Peshawar, Peshawar, Pakistan
                Universita degli Studi di Parma, ITALY
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0002-0813-2732
                Article
                PONE-D-18-14556
                10.1371/journal.pone.0209188
                6314602
                30601870
                d3ffbd2e-beb1-41ff-8942-3a3ebd262df4
                © 2019 Khan et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 7 June 2018
                : 30 November 2018
                Page count
                Figures: 4, Tables: 5, Pages: 17
                Funding
                The authors acknowledge the financial support provided by Pakistan Science Foundation (PSF) for research facilities. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Organisms
                Eukaryota
                Protozoans
                Parasitic Protozoans
                Cryptosporidium
                Biology and Life Sciences
                Organisms
                Eukaryota
                Protozoans
                Cryptosporidium
                Earth Sciences
                Hydrology
                Surface Water
                Biology and Life Sciences
                Parasitology
                Parasite Groups
                Apicomplexa
                Oocysts
                Biology and Life Sciences
                Organisms
                Eukaryota
                Protozoans
                Parasitic Protozoans
                Cryptosporidium
                Cryptosporidium Parvum
                Biology and Life Sciences
                Organisms
                Eukaryota
                Protozoans
                Cryptosporidium
                Cryptosporidium Parvum
                Medicine and Health Sciences
                Parasitic Diseases
                Protozoan Infections
                Biology and Life Sciences
                Molecular Biology
                Molecular Biology Techniques
                Artificial Gene Amplification and Extension
                Polymerase Chain Reaction
                Research and Analysis Methods
                Molecular Biology Techniques
                Artificial Gene Amplification and Extension
                Polymerase Chain Reaction
                Medicine and Health Sciences
                Gastroenterology and Hepatology
                Diarrhea
                Medicine and Health Sciences
                Diagnostic Medicine
                Signs and Symptoms
                Diarrhea
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Signs and Symptoms
                Diarrhea
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Animal Types
                Domestic Animals
                Biology and Life Sciences
                Zoology
                Animal Types
                Domestic Animals
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

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