Biochemical characterization and identification of bacterial strains isolated from drinking water sources of Kohat, Pakistan Tassadaq Hussain, Aneela Roohi, Shehzad Munir, Iftikhar Ahmed, Jafar Khan, Veronique Edel-Hermann, Kil Yong Kim, Muhammad Anees To cite this version: Tassadaq Hussain, Aneela Roohi, Shehzad Munir, Iftikhar Ahmed, Jafar Khan, et al.. Biochemical characterization and identification of bacterial strains isolated from drinking water sources ofKohat, Pakistan. African Journal of Microbiology Research, Academic Journal, 2013, 7 (16), pp.1579-1590. 10.5897/AJMR12.2204. hal-01005074 HAL Id: hal-01005074 https://hal.archives-ouvertes.fr/hal-01005074 Submitted on 29 May 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. 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Vol. 7(16), pp. 1579-1590, 16 April, 2013 DOI: 10.5897/AJMR12.2204 ISSN 1996-0808 ©2013 Academic Journals African Journal of Microbiology Research http://www.academicjournals.org/AJMR Full Length Research Paper Biochemical characterization and identification of bacterial strains isolated from drinking water sources of Kohat, Pakistan Tassadaq Hussain1, Aneela Roohi1, Shehzad Munir1, Iftikhar Ahmed2, Jafar Khan1, Veronique Edel-Hermann3, Kil Yong Kim4 and Muhammad Anees1* 1Department of Microbiology, Kohat University of Science and Technology, Kohat, Pakistan. 2SSO, Plant Biotechnology Program, National Institute for Genomics and Advanced Biotechnology (NIGAB), National Agricultural Research Center (NARC), PARC Road, Islamabad, Pakistan. 3INRA-Université de Bourgogne, UMR 1229 Microbiologie du sol et de l’environnement - CMSE, INRA, 17 rue Sully, BP 86510, 21065 Dijon, France. 4Division of Applied Bioscience and Biotechnology, Institute of Environmentally-Friendly Agriculture, Chonnam National University, Gwangju, 500-757, Korea. Accepted 25 February, 2013 As the pure drinking water is inevitable for good health, it is necessary to evaluate it for bacterial contamination. This study was conducted on the different drinking water sources of Kohat, a Northern- Western District of Pakistan. Sampling was done from different drinking water sources including tap water, tube well water, home-well, bore-well and springs. Physiochemical analyses including pH, temperature, turbidity, total dissolved solids and electrical conductivity showed that all water samples were within ranges of the values prescribed by the World Health Organization. A total of 79 bacteria isolated from different samples were characterized. Eighty two percent of the strains were Gram negative and 64% of the total Gram positive bacteria were spore forming. The physiological characterization showed that 30.4% of the total bacterial strains were obligate aerobes while the rest were facultative anaerobes. Biochemical characterization and identification depicted enormous bacterial diversity where sixteen genera could be tentatively identified. Identification of 24 of the strains was further validated by using API 20E kit. Furthermore, the selected strains were analyzed for pH, temperature optimization and NaCl tolerance. Pseudomonas sp were the most abundant bacteria followed by Bacillus sp. Some of the coliform bacteria could also be identified which present a potential health hazard. Key words: API 20E, bacterial diversity, water-borne pathogens. INTRODUCTION The water availability has always been of great per day for the world population (Yassi et al., 2001). importance for life and for every living organism. It has a About 69% of fresh water worldwide is used for domestic life-sustaining role in welfare and growth of mankind purpose that is for drinking, cooking sanitation, 22% in (Kazmi, 2004). All life on the earth is dependent on water industries, 8% for irrigation. and so does many economic activities. Every human The quality of water is important to the health, social individual require about 2 litres of clean drinking water and economic well being of people. It is important to test per day and this amount reaches to almost 12 millions m3 the suitability of the quality of water for its use as drinking *Corresponding author: E-mail: [email protected]. 1580 Afr. J. Microbiol. Res. water. Water that looks drinkable can contain bacterial Enterobacteriaceae example Escherichia coli, contamination, which are not visible to naked eye and Enterobacter aerogenes, Salmonella and Klebsiella spp. cannot be detected by smell, taste and sight. Variety of These enteropathogenic bacteria in water are responsible bacteria may be present in water even which looks for a variety of diseases like cholera, typhoid,dysenteries, clearand tasted good may not necessarily be safe to bacillary dysentery, etc. in human and livestock (Ashbolt, drink. Due to anthropogenic interventions the water is 2004). getting polluted and thus causing negative effects in This research work was based on the ecological human lives and in natural equilibrium. The problems in analysis of the indigenous bacterial strains of drinking water sources have become one of the primary problems water from different sources of Kohat District located in of human lives. Due to the water borne diseases which the North Western Province (Khyber Pakhtoonkhwa) of result from inadequate water supply, hygiene and Pakistan using a large sampling strategy. The study sanitation, around 4 million people suffer from water focuses on the isolation and identification of bacterial borne diseases and 2.2 million people die from these strains by morphological, cultural, and biochemical diseases every year (UNICEF-WHO, 2008). The problem methods. is even severe in developing countries where generally the drinking water is untreated. The infants are the most MATERIALS AND METHODS vulnerable targets of these diseases. Bacteria constitute Site description one of the major contaminants of water (Suthar et al., 2009). They have been reported to persist even in the Kohat is a medium sized town with a population of around 0.5 extreme environmental conditions and oligotrophic million people according to the last census data in 1998. The conditions (Sigee, 2005). Moreover, many of the bacterial annual population growth rate is 3.25%. It is located in Khyber- species have the ability to make resistant survival Pakhtunkhwa Province of Pakistan at 33°35'13N 71°26'29E with an elevation of 489 metres (1,607 ft). The total area of the district is structures. 2,545 square kilometres (983 sq mi). The river Indus forms the A variety of techniques have been reported so far to eastern boundary of the district, which separates it from the evaluate the ecology of bacteria in drinking water. For the province of Punjab. Kohat Toi is a principal stream, which enters characterization of drinking water bacterial strains, from Hangu district and flowing to east and southeast, drains into common techniques are culture-based methods and river Indus. Source of water for drinking purpose is mainly municipal biochemical characterization (Keinanen et al., 2004). The water that is tube well water, while some people also use home well water and spring water in some locations. Kohat features a semi- phenotypic techniques include morphological, cultural, arid climate with very hot summers and mild winters. The mean and physiological analysis. In addition to this a variety of maximum temperature in summer is over 40°C (104°F) and the molecular tools are available. The biochemical mean minimum temperature is 25°C (77°F). The mean minimum characterization has been very useful for assessment of temperature during winter is 4°C (39°F) and maximum is 18.35°C the bacterial ecology as well as the identification. (65.03°F). The rainfall is received throughout the year. The Moreover, kits are available for reliable bacterial monsoon rain is received from May to October. August is the rainiest month, with an average of about 111 mm. The winter rain identification based on the biochemical analysis. occurs from November to April. The highest winter rainfall is Nonetheless, these tools are very effective to study the received in the month of March. The average annual rainfall is biodiversity of bacterial flora. For instance, the API-20E about 546 mm. system is suitable for the identification of enteric bacteria and provides a convenient method to inoculate and read Water sample collection and physiochemical characterization tests relevant to members of the family Enterobacteriaceae and non-fastidious, Gram-negative Sampling was performed during the period of October (2010) to rods (Thaochan et al., 2010). API 20E identification March (2011) from different areas of District Kohat, Pakistan. A total system contains a strip of 20 miniaturized bio hanges, of 50 Samples were collected from wells, tube wells, spring, hand pumps, and tap water from different areas across the District Kohat and a database. Identification is obtained by the API 20E (Figure 1). Samples were collected aseptically in 500 ml sterile catalogue (Thaochan et al., 2010). The kit has been used autoclavable plastic (polypropylene) bottles and transferred in an in a number of studies for identification