Comparison of agar plate and real-time PCR on enumeration of Lactobacillus, Clostridium perfringens and total anaerobic bacteria in dog faeces

Letters in Applied Microbiology
C J FuM S Kerley

Abstract

To compare agar plate and real-time PCR methods on enumeration of total anaerobic bacteria, Lactobacillus and Clostridium perfringens in dog faeces. Thirty-two faecal specimens from Labrador retriever dogs were used to compare agar plate and real-time PCR enumeration methods for Lactobacillus, C. perfringens and total anaerobic bacteria. Total anaerobic bacteria, C. perfringens and Lactobacillus of faeces were counted (as CFU g(-1) faeces) for 48-h incubation at 37 degrees C in an anaerobic gas chamber on genus-selective media. Total genomic DNA from samples was extracted by the QIAamp DNA stool mini kit. The quantification of DNA (as DNA copy per gram faeces) by real-time PCR was performed with a LightCycler system with the QuantiTect SYBR green PCR kit for PCR amplification. The results indicated that there was a significant correlation between CFU and DNA copy of Lactobacillus (R2 = 0.78, P < 0.01) and total anaerobic bacteria (R2 = 0.21, P < 0.05); but no correlation was found between CFU and DNA copy of C. perfringens. The regression equations for Lactobacillus and total anaerobic bacteria were log(DNA copy) = 0.83 x log(CFU) + 1.43 and log(DNA copy) = 1.62 x log(CFU) - 6.32 respectively. The real-time PCR method could be ...Continue Reading

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Citations

Dec 15, 2010·Applied and Environmental Microbiology·Shu-Biao WuMingan Choct
Jul 31, 2012·Applied and Environmental Microbiology·Yurui TangPer E J Saris
Dec 18, 2009·International Journal of Microbiology·Hiroshi FukushimaYuko Akiba
May 17, 2014·Food & Function·Kyoung-Sik HanHarjinder Singh
Nov 30, 2018·Journal of the Science of Food and Agriculture·Chul Sang LeeSae Hun Kim
Mar 20, 2019·Scientific Reports·Fang OuFrédérique Vanholsbeeck
Aug 2, 2014·Asian-Australasian Journal of Animal Sciences·Yuanliang HuYunxiang Liang
Jan 6, 2015·Journal of Equine Science·Saori IshizakaHiroshi Matsuda
Jan 25, 2011·Letters in Applied Microbiology·K S HanM Boland
Aug 9, 2020·Frontiers in Microbiology·Paula Rodrigues Oblessuc, Maeli Melotto
Feb 6, 2019·Food Research International·Ju Young EorSae Hun Kim
Apr 4, 2021·Physiology & Behavior·Suparna Roy SarkarSugato Banerjee

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