An Investigation on the Bacterial Contents of Natural Springs in a Rural Area of the Middle Blacksea Region in Turkey

Alaa Aqso¹, Kübra Özkul¹ and Haydar Karakaya^1,

¹Department of Biology, Faculty of Science and Arts, University of Ondokuz Mayıs, Samsun, Turkey

Cite this paper:
Alaa Aqso, Kübra Özkul and Haydar Karakaya. An Investigation on the Bacterial Contents of Natural Springs in a Rural Area of the Middle Blacksea Region in Turkey. Applied Ecology and Environmental Sciences. 2014; 2(5):123-129. doi: 10.12691/aees-2-5-3

Abstract

Bacterial contamination levels of representative 4 natural springs were detected seasonally through indicator bacterial contents in a rural area of Ordu, Türkiye. Total coliform bacteria, enterococi, Escherichia coli and Clostridium perfringens spores were used as the indicator organisms. According to the bacterial parameters, none of the detected springs comply with the standards implemented by the state health authorities. Contamination level of the springs increases with human and domestic animal activities in spring and summer. Using PCR and duplex PCR, samples collected from the most densely contaminated spring, Soğuksu, were analyzed for three pathogenic bacterial groups. After eight-hour enrichment of the samples, positive results were found only for Salmonella spp. In conclusion, this investigation revealed that the natural springs used for drinking water supply in the research area are exposed to bacterial contaminations and the level of contamination increased with settlement density.

Keywords:
spring water bacterial contamination cultural tests PCR analyses

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Figures

References:

[1]	Chilton, J., Seiler, K.P., Groundwater occurrence and hydrogeological environments. In: Schmoll, O., Howard, G., Chilton, J., Chorus, I. (eds.) Protecting Groundwater for Health: Managing the Quality of Drinking-water Sources. WHO, IWA Publishing, London, 2006.
[2]	Howard, G., Bartram, J., Pedley, S., Schmoll, O., Chorus, I., Berger, P., Groundwater and public health. In: Schmoll, O., Howard, G., Chilton, J., Chorus, I. (ed.) Protecting Groundwater for Health: Managing the Quality of Drinking-water Sources. WHO, IWA Publishing, London. 2006.
[3]	Edwards, K.J., Becker, K., and Colwell, F., The Deep, Dark Energy Biosphere: Intraterrestrial Life on Earth. Annu. Rev. Earth Planet. Sci. 2012. 40:551-68
[4]	Reith, F., Life in deep subsurface. Geology 39:287-288. 2011.
[5]	Kleinsteuber, S., Schleinitz, K.M., Key players and team play: anaerobic microbial communities in hydrocarbon-contaminated aquifers. Appl Microbiol Biotechnol 94: 851-873. 2012.
[6]	Rheinheimer, G. Aquatic Microbiology. 4th edn. John Wiley&Sons, Chiester, 1991.
[7]	Brown, M.G., Balkwill, D.L., Antibiotic resistance in bacteria isolated from the deep terrestrial subsurface. Microbial Ecol 57:484-493. 2009.
[8]	Balkwill, D.L., Reeves, R.H., Drake, G.R., Reeves, J.Y., Croker, F.H., King, M.B., Boone, D.R., Pylogenetic characterization of bacteria in subsurface microbioal culture collections. FEMS Microbiol Rev 20: 201-216. 1997.
[9]	Roslev, P., Bukh, A.S,. State of the art molecular markers for fecal pollution source tracking in water. Appl Microbiol Biotechnol 89:1341-1355. 2011.
[10]	Macler, B.A., Merkle, J.C., Current knowledge on groundwater microbial pathogens and their control. Hydrogeol J 8:29-40. 2000.
[11]	Foppen, J.W., Schijven, J.F., Evaluation of data from the literature on the transport and survival of Escherichia coli and thermotolerant coliforms in aquifers under saturated conditions. Water Res 40(3):401-26. 2006.
[12]	Pedley, S., Yates, M., Schijven, J.F., West, J., Howard, G., Barrett, M., Pathogens: Health relevance, transport and attenuation. In Schmoll, O., Howard, G., Chilton, J., Chorus, I. (ed.). Protecting Groundwater for Health: Managing the Quality of Drinking-water Sources. WHO, IWA Publishing, London. 2006.
[13]	Gracias, K.S, McKillip, J.L., A review of conventional detection and enumeration methods for pathogenic bacteria in food. Can J Microbiol 50: 883-890. 2004.
[14]	Medema, G.J., Payment, P., Dufour, A., Robertson, W., Waite, M., Hunter, P., Kirby, R., Andersson, Y., Safe drinking water: An ongoing challenge. In: Assessing Microbial Safety of Drinking Water. OECD and WHO, IWA Publishing, London. 2003.
[15]	Stevans, M., Ashbolt, N., Cunliffe, D., Recommendations to change the use of coliforms as microbial indicators of drinking water quality, Australian Government, Health and Medical Research Council. Biotext Pty Ltd, Canberra. 2003.
[16]	Bartram, J., Pedley, S., Microbiological Analyses. In: Bartram, J.and Ballance, R. (ed), Water Quality Monitoring - A Practical Guide to the Design and Implementation of Freshwater Quality Studies and Monitoring Programme. UNEP/WHO, 229-255. 1996.
[17]	Leclerc, H., Mossel, D.A.A., Edberg, S.C., Struijk, C.B., Advances in the bacteriology of the coliform group: Their suitability as markers of microbial water safety. Annu Rev Microbiol 55:201-34. 2001.
[18]	Szewzyk, U., Szewzyk, R., Manz, W., Schleifer, K.H., Microbiological safety of drinking water. Annu Rev Microbiol 54:81-127. 2000.
[19]	Payment, P., Franco, E., Clostridium perfringens and somatic coliphages as indicators of the efficiency of drinking water treatment for viruses and protozoan cysts. Appl Environ Microbiol 59(8):2418-2424. 1993.
[20]	Salem, I.B., Aouni, M., Mzoughi, R., Two five-plex PCRs methods for identification of common Salmonella spp. Serotypes. Ann Microbiol 60:135-141. 2010.
[21]	Purohit, H.J., Kapley, A., PCR as an emerginig option in the microbial quality control of drinking water. Trends Biotechnol 20: 325-326. 2002.
[22]	Noble, R.T., Weisberg, S.B., A review of technologies for rapid detection of bacteria in recreational waters. J Water Health 03-4: 381-392. 2005.
[23]	Yamazaki, Y., Fukasawa, A., Multiplex polymerase chain reaction method discriminating Escherichia coli and Shigella sp. Arch Microbiol 193:83-87. 2011.
[24]	Kubista, M., Andrade, J.M., Bengtsson, M., Forootan, A., Jona, J., Lind, K., Sindelka, R., Sjoback, R., Sjogreen, B., Strombom, L., Stahlberg, A., Zoric, N., The real-time polymerase chain reaction. Mol Aspects of Med 27:95-125. 2006.
[25]	Arya, M., Shergill, I.S., Williamson, M., Gommersall, L., Arya, N., Patel, H.R.H. Basic principles of real-time quantitative PCR. Expert Rev Mol Diagn 5(2):2009-2019. 2005.
[26]	Henegariu, O., Heerema, N.A., Dlouhy, S.R, Vance, G.H, Vogt, P.H., Multiplex PCR: Critical parameters and step-by-step protocol. BioTechniques 23:504-511. 1997.
[27]	Heid, C.A., Stevens, J., Livak, K.J., Williams, P.M., Real time quantitative PCR. Genome Res 6: 986-994. 1996.
[28]	Park, S.H., Hanning, I., Jarauin, R., Moore, P., Donoghue, D.J., Donoghu, A.M., Ricke, S.C., Multiplex PCR assay for the detection and quantification of Campylobacter spp., Escherichia coli O157:H7 and Salmonella serotypes in water samples. FEMS Microbiol Lett 316: 7-15. 2011.
[29]	Kapley, A., Lampel, K., Prohit, H.J., Rapid detection of Salmonella in water samples by multiplex polimerase chain reaction. Water Environ Res 73(4):461-465. 2001.
[30]	Smith, C.J., Osborn, A.M., Advantages and limitations of quantitative PCR(Q-PCR) based approaches in microbial ecology. FEMS Microbiol Ecol 67:6-20. 2009.
[31]	Bej, A.K., Steffan, R.J., DıCesare, J., Haff, L., Atlas, R.M., Detection of coliform bacteria in water by polymerase chain reaction and gene probes. Appl Environ Microbiol 56(2): 307-314. 1990.
[32]	TS EN ISO 9308-1, Su kalitesi- Escherichia coli ve Koliform bakterilerin tespiti ve sayımı. Bölüm 1: Membranla süzme yönetemi. Türk Standardları Enstitüsü, Ankara. 2004.
[33]	Chapman, G.H., A culture medium for detecting and confirming Escherichia coli in ten hours. Am J Public Health N, 41:1381-1381. 1951.
[34]	TS EN ISO 7899-2, Su kalitesi bağırsak enterokoklarının tespiti ve sayımı- Bölüm 2: membran süzme yöntemi. Türk Standardları Enstitüsü, Ankara. 2002.
[35]	TS EN ISO 8020, Su kalitesi- Sülfit indirgeyen havasız ortam (anaerob) bakteri (clostridia) sporlarının aranması ve sayımı. Türk Standardları Enstitüsü, Ankara. 1997.
[36]	Araujoa, M., Sueiro, R.A., Gomez, M.J., Garrido, M.J., Enumeration of Clostridium perfringens spores in groundwater samples: comparison of six culture media. J Microbiol Meth 57: 175-180. 2004.
[37]	Sambrook, J., Russell, D.W. Molecular cloning a laboratory manual. 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York. 2001.
[38]	Sharma, V.K., Dean-Nystrom, E.A., Casey, T.A., Semi-automated fluorogenic PCR assays (Taqman) for rapid detection of Escherichia coli O157:H7 and other shiga toxigenic E. coli. Mol Cell Probes 13: 291-302. 1999.
[39]	Cheng, C.M, Lin, W., Van, K.T, Phan, L., Tran, N.N, Farmer, D., Rapid detection of Salmonella in foods using real-time PCR. J Food Protect 71: 2436-2441. 2008.
[40]	Ruxton, G.D., The unequal variance t-test is an underused alternative to Student’s t-test and the Mann–Whitney U test. Behav Ecol 686-690. 2006.
[41]	Ordu Halk Sağlığı Müdürlüğü, Ordu İli 2012 Yılına Ait İçme Suyu Analiz Sonuçları. 2013. [Online]. Available: (http://www.orduhalksagligi.gov.tr/wp/?page_id=353 07.07.2013).
[42]	Sağlık Bakanlığı, İnsani tüketim amaçlı sular hakkında yönetmelik. Resmi Gazete, Sayı:25730. 17 Şubat 2005.