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Comparison of Multiple Tube Fermentation and Polymerase Chain Reaction Methods for the Detection of Coliforms in Freshwater Publisher



Fard RMN1, 2 ; Asgari FS3 ; Ashrafi I4 ; Gharibi S5 ; Khani MR6
Authors
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Authors Affiliations
  1. 1. Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Food Microbiology Research Center, Tehran University of Medical Sciences, Tehran, Iran
  3. 3. Department of Veterinary Theriogenology and Obstetrics, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
  4. 4. Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
  5. 5. Jahad Daneshgahi of the University of Tehran, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
  6. 6. Department of Environmental Health Engineering, Faculty of hygiene, Islamic Azad University, Tehran Medical Sciences, Tehran, Iran

Source: Applied Food Biotechnology Published:2019


Abstract

سببقه و هذف: ض اٌسایی هیکز بٍ اّی تیواریشا یکی اس هسایل ه نْ ت ذْاضت آب ت ضوار هی آیذ. گ اًَِّای اضزضایا کلی ت ػ اٌَى طًا گًز آل دَگی هذف ػَی در آب ه رَد استفاد قزار هی گیز ذً. در ایاي هاالؼا ،ِ هیاشاى کاارایی د ر شٍ تخویز چ ذٌ ل لَ اِی اٍک صٌ س دًیز ای پلیوزاس در ض اٌسایی کلیفزم اّ )ت یٍض اضزضیا کلی( در آب ه رَد هقایس قزار گزفت ا ذً. مواد و روش هب: ت ه ظٌ رَ هقایس ر شٍ اّی تخویز چ ذٌ ل لَ اِی اٍک صٌ س دًیز اُی پلیوزاس، 15 وً آب اس پا حٌ ه ثٌغ گ اًَگ ىَ )س وً اس آب ر دٍخا ،ًِ ضص وً اس آب چا ،ُ سا وً اًَ اس آب ضایز سا وً اًَ اس آب تاازی( خوغآ رٍی ضذ. وً اًَِّ تز هحیط کطت لاکت سَ تزاث ت ه ظٌ رَ تؼییي تیطتزیي ضوارش احتوالی تاکتزیایی کطت داد ضذ ذً. ػلا تزایي، تزای تزرسی صى اّی lacZ )هؼزف حض رَ کلیفزم( uidA )هؼزف حضا رَ اضزضایا کلای ( اس ر شٍ فیلتز استات سل لَش تزای استخزاج DNA تاکتزیایی استفاد ضذ. ) 53 ( ضاص ) 40 / یبفته هب و نتیجهگیری: تز اساط تًایح ت دست آهذ اس ر شٍ خویز چ ذٌ ل لَا ای، طّات ) 3 وً اًَِّ ت تزتیة ت کلیفزم اضزضیا کلی آل دَگی ت دَ ذً. ػلا تز ایي، تًایح ر شٍ اٍک صٌ س دًیز اُی پلیوازاس طًااى 53 ( وً اًَِّ ت تزتیة ت کلیفزم اضزضیا کلی آل دَ ت دَ ذً. تًایح طًاى هید ذّ ک / 66 ( طّت ) 3 / داد ک د ) 7 تزای تطخیص آل دَگی آب ت کلیفزم، ر شٍ اٍک صٌ س دًیز اُی پلیوزاس تسیار سزیؼتز، دقیا تاز حسااط تاز اس ر شٍ س تٌی )ها ذًٌ ر شٍ خویز چ ذٌ ل لَ اِی( هیتاضذ. ت ػلا ،ٍُ تا ر شٍ اٍکا صٌ چ ذٌگا اً س دًیاز ای پلیوازاس ) M-PCR ) چ ذٌیي عًَ تاکتزی ت ط رَ وّشهاى قاتل ردیاتی هیتاض ذٌ. تعبرض منبفع: یًَس ذٌگاى اػلام هی ک ذٌٌ کِ یّچ عًَ تؼارض ه اٌفؼی هزتثط تا ا تًطار ایي هقالِ ذًار ذً.; Background and objective: Detection of microbial pathogens in water is one of the major health issues. Escherichia coli species are used as indicators of fecal contamination in water microbial detection. In this study, efficacies of two methods of multiple tube fermentation and polymerase chain reaction have been compared for the detection of coliforms (especially Escherichia coli) in water. Material and methods: To compare multiple tube fermentation and polymerase chain reaction methods, 15 water samples were collected from five different sources (three gutter, six well, three tap and three bottled mineral water samples). The samples were cultured in lactose broth media to achieve the most probable number of bacteria. Furthermore, acetate cellulose filter method was used for the bacterial DNA extraction to investigate lacZ (indicating the presence of coliforms) and uidA (indicating the presence of Escherichia coli) genes. Results and conclusion: Based on the results of multiple tube fermentation, eight (53.3%) and six (40%) samples were contaminated with coliforms and Escherichia coli, respectively. Furthermore, polymerase chain reaction results showed that ten (66.7%) and eight (53.3%) samples contained coliforms and Escherichia coli, respectively. Results have suggested that polymerase chain reaction is much faster, more accurate and more sensitive than traditional methods (e.g. multiple tube fermentation) for the detection of coliform contaminated water. Moreover, several types of bacteria can be tracked simultaneously by M-PCR. Conflict of interest: The authors declare no conflict of interest. © 2019 Applied Food Biotechnology.