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      Microbial Groundwater Quality Status of Hand-Dug Wells and Boreholes in the Dodowa Area of Ghana

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          Abstract

          To assess the suitability of water sources for drinking purposes, samples were taken from groundwater sources (boreholes and hand-dug wells) used for drinking water in the Dodowa area of Ghana. The samples were analyzed for the presence of fecal indicator bacteria ( Escherichia coli) and viruses (Adenovirus and Rotavirus), using membrane filtration with plating and glass wool filtration with quantitative polymerase chain reaction (PCR), respectively. In addition, sanitary inspection of surroundings of the sources was conducted to identify their vulnerability to pollution. The presence of viruses was also assessed in water samples from the Dodowa River. More than 70% of the hand-dug wells were sited within 10 m of nearby sources of contamination. All sources contained E. coli bacteria, and their numbers in samples of water between dug wells and boreholes showed no significant difference ( p = 0.48). Quantitative PCR results for Adenovirus indicated 27% and 55% were positive for the boreholes and hand-dug wells, respectively. Samples from all boreholes tested negative for the presence of Rotavirus while 27% of the dug wells were positive for Rotavirus. PCR tests of 20% of groundwater samples were inhibited. Based on these results we concluded that there is systemic microbial and fecal contamination of groundwater in the area. On-site sanitation facilities, e.g., pit latrines and unlined wastewater drains, are likely the most common sources of fecal contamination of groundwater in the area. Water abstracted from groundwater sources needs to be treated before use for consumption purposes. In addition, efforts should be made to delineate protected areas around groundwater abstraction points to minimize contamination from point sources of pollution.

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          Rapid and quantitative detection of human adenovirus DNA by real-time PCR.

          Rapid diagnosis of human adenovirus (HAdV) infections was achieved by PCR in the recent years. However, conventional PCR has the risk of carry-over contamination due to open handling with its products, and results are only qualitative. Therefore, a quantitative "real-time" PCR with consensus primer and probe (dual fluorescence labelled, "TaqMan") sequences for a conserved region of the hexon gene was designed and evaluated. Real-time PCR detected all 51 HAdV prototypes. Sensitivity of the assay was
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            Sanitation and Health

            As one article in a four-part PLoS Medicine series on water and sanitation, David Trouba and colleagues discuss the importance of improved sanitation to health and the role that the health sector can play in its advocacy.
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              Rapid typing of human adenoviruses by a general PCR combined with restriction endonuclease analysis.

              We have developed a system for rapid typing of adenoviruses (Ads) based on a combination of PCR and restriction endonuclease (RE) digestion (PCR-RE digestion). Degenerated consensus primers were designed, allowing amplification of DNA from all 51 human Ad prototype strains and altogether 44 different genome variants of Ad serotypes 1, 3, 4, 5, 7, 11, 19, 40, and 41. The 301-bp amplimer of 22 prototype strains representing all six subgenera and the genome variant was selected as a target for sequencing to look for subgenus and genome type variabilities. The sequences obtained were used to facilitate the selection of specific REs for discrimination purposes in a diagnostic assay by following the concept of cleavage or noncleavage of the 301-bp amplimer. On the basis of these results, a flowchart was constructed, allowing identification of subgenus B:2 and D serotypes and almost complete distinction of subgenus A, B:1, C, E, and F serotypes. Application of the PCR-RE digestion system to clinical samples allowed typing of 34 of 40 clinical samples positive for Ad. The genome type determined by this method was identical to that obtained by traditional RE typing of full-length Ad DNA. The remaining six samples were positive only after a nested PCR. Therefore, to reduce the risk of false-negative results, samples scored negative by the PCR-RE digestion system should be evaluated by the described nested PCR. Used in combination, the PCR-RE digestion method and the nested PCR provide a reliable and sensitive system that can easily be applied to all kinds of clinical samples when rapid identification of adenoviruses is needed.
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                Author and article information

                Journal
                Int J Environ Res Public Health
                Int J Environ Res Public Health
                ijerph
                International Journal of Environmental Research and Public Health
                MDPI
                1661-7827
                1660-4601
                12 April 2018
                April 2018
                : 15
                : 4
                : 730
                Affiliations
                [1 ]Department of Civil Engineering, Central University, Miotso-Campus, Miotso Tema, Ghana; glutterodt@ 123456central.edu.gh
                [2 ]Department of Environmental Engineering and Water Technology, IHE Delft Institute for Water Education, Westvest 7, 2611 AX Delft, The Netherlands; kolif.02@ 123456gmail.com
                [3 ]Department of Water Science and Engineering, IHE Delft Institute for Water Education, Westvest 7, 2611 AX Delft, The Netherlands; y.hoiting01@ 123456gmail.com (Y.H.); j.foppen@ 123456un-ihe.org (J.W.A.F.)
                [4 ]Department of Civil Engineering, Kwame Nkrumah University of Science and Technology (KNUST), Kumasi, Ghana; sokwarteng@ 123456gmail.com
                Author notes
                [†]

                Shared first authorship.

                Author information
                https://orcid.org/0000-0003-4497-6155
                Article
                ijerph-15-00730
                10.3390/ijerph15040730
                5923772
                29649111
                d4e04036-8616-4239-b057-8717124cd8db
                © 2018 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 19 February 2018
                : 10 April 2018
                Categories
                Article

                Public health
                groundwater quality,adenovirus,rotavirus,escherichia coli
                Public health
                groundwater quality, adenovirus, rotavirus, escherichia coli

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