The Research Of Using Faecalibaterium To Trace The Source Of Faces Pollution In Ground Water

Traditional tracking microbes, such as E.coli, Enterococcus spp, Bifidobacterium,Clostridium perfringens, Prevotella, and animal intestinal virus, have many limiationsfor water tracking detection, and it is difficult to trace the degree of water pollution. thelatest study shows that: Faecalibacterium is a kind of dominant bacteria in human andanimal’s intestinal, and Faecalibacterium in different host intestinals have significantdifferences in gene. According to these differences in the gene can distinguish fecalsources is a great advantage which can greatly make up for the defects of traditionalfecal pollution tracking microbes.Through the design of a pair of universal primers FaU-F and FaU-R to detectFaecalibacterium in six kinds animals as human, pig, cow and sheep, chicken andduck’s fecal, The experimental results show: the genomic DNA samples of six kinds ofanimal facel amplified fragments of the same size, the fragment size is500bp, whichmeans the human and animal intestinal bacteria Faecalibacterium is widespreadcharacteristics.According to the Faecalibacterium in human intestinal to design a pair of primersHFB-F3and HFB-R5to do mutiply PCR amplifications of genomic DNA in humanfacel, the results showe: the optimum PCR reaction conditions are as follows: theannealing temperature is55℃, primer concentration was10μ mol/L, the template was50ng, the time of amplification cycles is35. Using HFB-F3and HFB-R5primers todetect the genomic DNA of human fecal,6of the10facel samples amplify the targetband, the fragment size is399bp, positive amplification rate is60%. Choosing thegenomic DNA of the other five animals chicken, duck, pig, dog, and cow for thespecific control test, all the samples produce no band.According to the Faecalibacterium in chicken intestinal to design two pairs ofprimers FaCH-F1, FaCH-R1and FaCH-F2, FaCH-R2for the detection of chicken fecalpollution, the results show: the first pair of primers amplifies the target band of97bpfor40%chickens; the first pair of primers amplifies the target band of132bp for70%chickens. Which means primers II are more sensitive than primers I, using the two pairsof primers both can track Faecalibacterium in chicken fecal can get the object to detectthe quality of water polluted by chicken facel.Through gene cloning and sequencing have identified the conserved sequence of 16S rDNA in duck intestinal Faecalibacterium, and design the PCR primers foramplifing the duck intestinal Faecalibaterium16S rDNA. Using the primers toconstruct the16S rDNA sequence library of Faecalibaterium in duck intestinal. Andfind four specific sequence fragments in200clone samples, and design four specificprimers.This paper shows the Faecalibacterium’s advantage as a kind of water fecalpollution indicator bacteria, do the preliminary study for the detection method ofFaecalibacterium in aquatic envirenment, the method can distinguish the sources ofdifferent facels, has good stability, and wide application prospects.