| The huge amounts of microbial flora in human intestinal and host constitute the microecological balance which ensure life activities of host work normally. If this balance is destroyed, host will be to get some diseases easily. Many study suggested that a lot of factors including internal and external can play a important role in the diversity and formation of intestinal flora. Especially, the relationship between intestinal flora and host age has not expounded yet, and the host age is one of key factors which defines the structure of the intestinal flora. So we collected 28 volunteers of different age from the world’s longevity township BaMa Town in guangxi and applied the combination of microbial pure culture method, DGGE technology and RT-PCR technology to study the age factor influence on intestinal flora from guangxi and provide theoretical basis for the correlation of intestinal flora and human age. The main results are as following:1. A total of 28 stool specimens were collected in three longevity village of Bama and using selective mediums to separate the lactic acid bacteria and analyse their quantity and tolerance and the bacteriostatic ability. According to age, sample can be divided into four groups:youth, middle-aged, elderly group and longevity group. The results showed that 1273 strains were isolated from those samples. According to corresponding physiological and biochemical tests, 1178 strains were lactic acid bacteria. The number of lactic acid bacteria in youth group(9.16±0.181gcfu/g) was higher than the other groups, as the growth of the age, the number of lactic acid bacteria reduced gradually, but the longevity group(8.89±0.481gcfu/g) of lactic acid bacteria number still kept a high level; the lactic acid bacteria in middle-aged(40.85±3.27%) and elderly group(37.26±2.32%) tolerance ability of pH 3 gastric juice were similar and were significantly greater than the youth and longevity group(P<0.05), especially the longevity group(28.26±3.85%) was worst; lactic acid bacteria in different age groups had no significant difference in intestinal 0.5% of juice bile salt and pH 3 intestinal juice resistant ability and tolerance ability; the inhibitory effect on E.coil of youth and middle-aged group was significantly higher than elderly and longevity group, diameter of antibacterial circle were 18.67±0.98mm, 17.98±0.71mm, the inhibitory effect on Staphylococcus aureus of longevity group was lower than youth and middle-aged group, diameter of antibacterial circle was 15.41±1.48mm.2. PCR-DGGE technology was applied to study intestinal flora in different ages group comparatively. The results showed that for advantage bacterium group, expect middle-aged group, the affinity of the rest three groups of was low, but had high affinity withn groups, the main different strains were Bacteroides vulgatus, Ruminococcus, Bifidobacterium adolescentis, Prevotella melaninogenica, Clostridiales, Acinetobacter baumannii, Staphylococcus epidermidis, Lactobacillus casei, Eubacterium eligens, Escherichia fergusonii, Uncultured bacterium; for bacteroides, the affinity between youth and longevity group was high, had low affinity with other groups, the affinity between middle-aged and elderly group was high, and had high affinity withn youth, middle-aged and longevity group, low affinity withn elderly group, the main different strains were Bacteroides vulgatus, Bacteroides salanitronis, Bacteroides thetaiotaomicron, Parabacteroides distasonis, Tannerella forsythia, Bacteroides fragilis, Uncultured bacterium; for Clostridium, the affinity between youth and longevity group was high, had low affinity with other groups, the affinity between middle-aged and elderly group was low, and low high affinity withn youth and middle-aged, high affinity withn longevity group and elderly group, the main different strains were Ruminococcus champanellensis, Ruminococcus albus, Ruminococcus torques, Clostridium cellulolyticum, Clostridiales genomosp., Ruminococcus bromii, Clostridium phytofermentans, Clostridium pasteurianum, Clostridium saccharolyticum, Clostridium cellulolyticum, Clostridium acidurici, Clostridium saccharobutylicum, Uncultured bacterium; for Bifidobacteria, had high affinity between four groups, but each group had one sample different others, the main different strains were Bifidobacterium bifidum, Bifidobacterium thermophilum, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium animalis, Bifidobacterium dentium, Bifidobacterium adolescentis, Uncultured Bifidobacterium sp.;for Lactobacillus, the affinity between youth and longevity group was high, had low affinity with other groups, the affinity between middle-aged and elderly group was high, but each group had one sample different others, the main different strains were Lactobacillus plantarum, Lactobacillus salivarius, Lactobacillus ruminis, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus acidophilus, Lactobacillus buchneri, Lactobacillus reuteri, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii.3. RT-PCR technology was applied to study intestinal flora in different age groups. The results showed that for Enterobacteriaceae, the number of middle-aged group (8.16±0.29 lgcopies/g) was slightly higher than the longevity(7.93±0.571gcopies/g) and elderly group(7.89±0.421gcopies/g), significantly higher than youth group(7.46±0.321gcopies/g, P<0.05); for Enterococcus, the number of middle-aged group(7.06±0.44 lgcopies/g) was slightly higher than elderly group(7.89±0.331gcopies/g), significantly higher than youth(7.06±0.441gcopies/g) and longevity group(6.93±0.371gcopies/g, P＜0.05); for Clostridium, the number of youth(9.06±0.441gcopies/g) and middle-aged group(8.91±0.171gcopies/g) were significantly higher than elderly(8.42±0.231gcopies/g) and longevity group(8.13±0.261gcopies/g, P<0.05); for Lactobacillus, the number of longevity group(8.63±0.381gcopies/g) was slightly higher than youth group(8.36±0.481gcopies/g), significantly higher than middle-aged(7.85±0.211gcopies/g) and elderly group(7.98±0.221gcopies/g, P＜0.05); for Bacteroides, the number of youth group(9.76±0.351gcopies/g) was slightly higher than longevity group(9.53±0.281gcopies/g), significantly higher than middle-aged(9.23±0.171gcopies/g) and elderly group(8.92±0.32 lgcopies/g,P<0.05); for Bifidobacteria, the number of longevity group(10.52±0.391gcopies/g) was slightly higher than youth group(10.05±0.461gcopies/g), significantly higher than middle-aged(8.86±0.411gcopies/g) and elderly group(8.33±0.921gcopies/g, P＜0.05). The number of Bifidobacteria was slightly higher than Bacteroides, significantly higher than other four dominant bacteria(P<0.05).
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