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Research Article Identification of Opportunistic Pathogenic Bacteria in Drinking Water Samples of Different Rural Health Centers and Their Clinical Impacts on Humans

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Research Article Identification of Opportunistic Pathogenic Bacteria in Drinking Water Samples of Different Rural Health Centers and Their Clinical Impacts on Humans Hindawi Publishing Corporation BioMed Research International Volume 2013, Article ID 348250, 10 pages http://dx.doi.org/10.1155/2013/348250 Research Article Identification of Opportunistic Pathogenic Bacteria in Drinking Water Samples of Different Rural Health Centers and Their Clinical Impacts on Humans Pavan Kumar Pindi, P. Raghuveer Yadav, and A. Shiva Shanker Department of Microbiology, Palamuru University, Mahabubnagar, Andhra Pradesh 509001, India Correspondence should be addressed to Pavan Kumar Pindi; [email protected] Received 1 March 2013; Revised 22 April 2013; Accepted 24 April 2013 Academic Editor: Pedro Cabrales Copyright © 2013 Pavan Kumar Pindi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. International drinking water quality monitoring programs have been established in order to prevent or to reduce the risk of contracting water-related infections. A survey was performed on groundwater-derived drinking water from 13 different hospitals in the Mahabubnagar District. A total of 55 bacterial strains were isolated which belonged to both gram-positive and gram-negative bacteria. All the taxa were identified based on the 16S rRNA gene sequence analysis based on which they are phylogenetically close to 27 different taxa. Many of the strains are closely related to their phylogenetic neighbors and exhibit from 98.4 to 100% sequence similarity at the 16S rRNA gene sequence level. The most common group was similar to Acinetobacter junii (21.8%) and Acinetobacter calcoaceticus (10.9%) which were shared by 7 and 5 water samples, respectively. Out of 55 isolates, only 3 isolates belonged to coliform group which are Citrobacter freundii and Pantoea anthophila. More than half (52.7%, 29 strains) of the phylogenetic neighbors which belonged to 12 groups were reported to be pathogenic and isolated from clinical specimens. Out of 27 representative taxa are affiliated have eight representative genera in drinking water except for those affiliated with the genera Exiguobacterium, Delftia, Kocuria, and Lysinibacillus. 1. Introduction access to usual safe drinking water and several others are threatened [1]. Human population growth wields several and various pres- Potable or drinking water is defined as having a sat- sures on the quality and the quantity of drinking and fresh isfactory quality in terms of its physical, chemical, and water resources and on the access to them. Safe drinking bacteriological parameters so that it can be securely used for water remains inaccessible to several millions people in drinking and cooking. The most common and widespread theglobe.Safedrinkingwater,thus,reducestheloadof health risks associated with drinking water in developing waterborne diseases. Contamination of drinking water due countries are of biological origin. The WHO estimates that to natural and manmade contaminants is frequently reported about 1100 million people globally drink unsafe water, and in developing countries where mainstream of the inhabitants the greater part of diarrheal disease in the globe (88%) is survive in countryside and uptown areas with meager hygiene attributable to insecure water, sanitation, and hygiene [2]. and waste clearance practices. In developing countries, poor Ten major waterborne diseases are responsible for over 28 water quality is the most important risk of child mortalities billion disease episodes annually in developing countries. which are mainly through infectious diarrhea. In India, about Thequalityofdrinkingwatermightbeascertainedbyits 10% of the countryside and city populations do not have microbiological examination. The maximum threat from 2 BioMed Research International Table 1: Bacterial abundance from the drinking water samples Hospitals drinking (potable) water systems are the most collected from government hospitals, Mahabubnagar. important and controllable as well as the most overlooked source of nosocomial pathogens. Conventional culture-based Serial Sample Place of sample Number of Isolated strains microbiological water quality monitoring techniques take a number number collection isolates long time (several days), and usually a small volume of water 1 AL Alampur 3 AL1, AL3, AL4 is sampled (typically 100 mL), which gives rise to inadequate 2 AP Achampet 3 AP1, AP2, AP4 detection limits with regard to drinking water safety. Further- D12, D22, D24, 3DShadnagar4 more, the presence of some important waterborne pathogens D25 (such as Pseudomonas aeruginosa or legionellae)showsno 4 GD Gadwal 4 GD2, GD4–D6 correlation with conventional indicator organism counts. 5 JD Jadcherla 4 JD1–JD4 Water-related pathogens can also find niches in water systems 6 JU Jurala 4 JU1–JU4 (i.e., an association with biofilms or free-living amoebae), rendering their observation with conventional techniques 7 KL Kalwakurthy 5 KL1–KL5 more difficult. Molecular techniques provide new and rapid 8 MB Mahabubnagar-1 4 MB1–MB4 facilities for the detection of pathogens involved in nosoco- 9 MK Makthal 3 MK2–MK4 mial infections. Five representative end-points (taps) in the 10 NG Nagar Kurnool 2 NG1, NG2 drinkingwatersystemofahospital(Budapest,Hungary)were 11 NR Narayanpet 3 NR2–NR4 sampled in October 2005. R1, R2, R5–R8, 12 R Mahabubnagar-2 12 R21–R23, R26, 2. Materials and Methods R30, R31 WN1, 2.1. Source of Samples. Thirteen drinking water samples were 13 WN Wanaparthy 4 WN3–WN5 collected from different government hospitals, Mahabubna- gar District, Andhra Pradesh, India, on 22nd and 23rd of November, 2009. Fifty five strains were isolated from these 13 ∘ drinking water sources on nutrient agar medium at 37 C. For isolation of bacteria, 100 L of the water sample was plated on ∘ microbes (coliform) in water is related to consumption of nutrient agar medium and incubated at 37 Cfor2days.Based drinking water, that is, contaminated with human or animal on the different colony morphology from each sample, a total excreta. Coliform bacteria include members of the family of 55 strains were selected and characterized in the present Enterobacteriaceae, for example, Escherichia coli, Enterobac- study. ter aerogenes, Salmonella, and Klebsiella in water that are accountable for a variety of diseases like cholera, typhoid, dysenteries, bacillary dysentery, and so forth in human and 2.2. 16S rRNA Gene Sequencing and Phylogenetic Analysis. livestock [3]. For 16S rRNA gene sequencing, DNA was prepared using the Mo Bio microbial DNA isolation kit (Mo Bio Laboratories Recently, Suthar et al. [4, 5] reported a wide range of Inc., Solano Beach, CA, USA) and sequenced as described pathogenic bacteria in potable water samples from some previously [6]. The resultant almost complete sequence of the rural habitations of Northern Rajasthan, India. A total of 16S rRNA gene contained 1502 nucleotides. The 16S rRNA ten bacterial species, E. coli, Pseudomonas aeruginosa, E. aer- gene sequence of the isolate was subjected to BLAST sequence ogenes, Klebsiella sp., Proteus vulgaris, Alcaligenes faecalis, similarity search [7]andEzTaxon[8] to identify the nearest Bacillus cereus, Staphylococcus aureus, Streptococcus lactis, taxa. The entire related 16S rRNA gene sequences were down- and Micrococcus luteus, were isolated and identified from loadedfromthedatabase(http://www.ncbi.nlm.nih.gov/) the potable water samples from this region [4] M. luteus, S. aligned using the CLUSTAL Xprogram[9]andthealign- lactis, Klebsiella sp., and E. coli were the dominant microflora ment corrected manually. Phylogenetic tree was constructed (recorded from 73.1% of the villages/towns) in the water using tree-making algorithm and the Maximum likelihood samples. The presence of coliforms shows the danger of (ML) using the PhyML program [10]. fecal pollution and a consequent hazard of contracting a disease through pathogenic organisms. Nonetheless, the disease-causing organisms (pathogens) mostly transmitted 2.3. Biochemical Tests. The biochemical tests were performed via drinking water are predominantly of fecal origin. for all the strains obtained by using the HiMViCTM Bio- The results of the recent study on the detection of chemical Test Kit (HIMEDIA-KB001) and HiAssorted Bio- potentially pathogenic bacteria in the drinking water dis- chemical Test Kit (HIMEDIA-KB002). tribution system of a hospital in Hungary by Felfoldi¨ et al. [5] had shown the presence of Legionella, Pseudomonas 3. Results and Discussion aeruginosa, and also several other opportunistic pathogenic bacteria, such as Escherichia albertii, Acinetobacter lwoffii 3.1. Characterization of the Bacterial Strains from Drinking and Corynebacterium tuberculostearicum,emphasizingthat Water. All these isolates were from different drinking water drinking water systems, particularly those with stagnant sources, and all are mesophilic and could grow in the temper- ∘ water sections, could be the source of nosocomial infections. ature range of 20 to 40 C with optimum growth temperature BioMed Research International 3 T Aquifex pyrophilus Kol5a (M83548) T 100 Kocuria rhizophila DSM 11926 (Y16264) Kocuria sp. R8(2010) () T 100 Exiguobacterium indicum HHS31 (AJ846291) Exiguobacterium sp. D25(2010) () T 100 Lysinibacillus fusiformis NBRC 15717 (AB271743) 100 Lysinibacillus sp. JU2(2010) (GU566326) T 81 Bacillus cereus ATCC 14579 (AE016877) 97 Bacillus sp. R5(2010) () 93 100 Bacillus sp. D12(2010) () T Bacillus anthracis ATCC 14578 (AB190217)
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