Quantity Of Desulfovibrio And Analysis Of Intestinal Microbiota Diversity In Different People

Sulphate-reducing bacteria (SRB) is a kind of anaerobic bacteria which can producthydrogen sulphide by reducting sulfate. Desulfovibrio (DSV) is the predominant SRB speciein human colonic microbiota. Because hydrogen sulphide is a potential aetiological agent toepithelial cells. Many researchers guessed that, there may be a link between DSV andintestinal diseases. But in the current studies, debate persists regarding the role of DSVsubspecies in intestinal diseases. Our research collected the intestinal contents of ulcerativecolitis (UC), colorectal cancer (CRC), polypus (PP) and healthy control (H) people from TheFourth People’s Hospital in Wuxi city, Jiangsu province. We provide an overview of thequantitive difference of DSV in human intestinal tract of different groups of people, and therelationship between DSV quantity and age, gender. Our research combined the diversity ofDSV and intestinal microbiota, to explore the potential relationship between DSV quantity,intestinal flora diversity and intestinal diseases. There are positive significance in preventingintestinal diseases, revealling the pathogenic mechanism and the development of drugs forintestinal diseases.In our research, we collected the intestinal contents of58subjects, and devided in fourgroups: CRC (15), PP (15), UC (13) and H (15). The total DNAwas extracted by Blood DNAkit, the DNA was used as template to amplificate specific gene fragment of DSV. Real timefluorescence quantitative PCR (RT-PCR) assay was used to enumerate the number of DSV inintestinal tract of58subjects, and analyze the relationship between DSV quantity and age,gender. Using the intestinal bacteria total DNA as template, we used Nested-PCR toamplificate GC-dsrB and GC-16S rRNA V3gene fragments. Diversity of DSV and gutmicrobiota were distinguished by analysis of dsrB gene and the V3region of16S rRNAgeneusing PCR-Denaturing Gradient Gel Electrophoresis (PCR-DGGE).DSV was detected using RT-PCR in all the samples. Significantly increased number ofDSV was observed for UC (2.9×106±1.6×105cfu/mL) and PP (1.2×106±8.6×105cfu/mL)groups compared with both CRC (6.8×105±1.0×105cfu/mL) and H (7.0×105±2.1×105cfu/mL)groups (p<0.05). No significant difference was observed for CRC and H groups with thenumber of DSV in intestinal content (p>0.05). There was no relationship between DSVquantity and age in four groups of people (p>0.05). In PP group, the quantity of intestinalDSV in male is higher than female (p<0.05), while in the other three groups have no obviousdifference (p>0.05). The analysis of DGGE profile and sequencing of16S rRNA V3geneshowed alteration of DSV and gut microbiota by comparing disease groups with control.With the combination of RT-PCR and DGGE, our research found that quantity anddiversity of DSV are significantly increased in UC and PP compared to control. The increasednumber of DSV in disease groups suggests a possible harmful role in intestinal health. Thediversity of gut microbiota in disease groups increased compared with controls as well. Also,there is significant alteration in the composition of gut microbiota in patients with UC, PP andCRC compared to control.