Identification Of Marine Fouling Microbe And Preliminary Study On The Its Specific Amplification Method

During the development process of ocean, marine fouling organisms attachmentis a very serious issue. Marine fouling microorganisms refer to those microbialpopulations that attach to the surfaces of the bottom, pipeline, buoy and artificialmarine structures of ships. For many years, fouling damages towards nationaldefense, shipping, coastal factory cooling systems, and aquatic products industryhave aroused a wide concern among domestic and foreign scholars. But thedynamics process and key steps of early fouling the microbialcommunity are stillobscure. In this study, systematic characterization was carried out to identify early-stage fouling microbes in the Xiaoshi Island area at Weihai, and based on the wholegenome sequences of determined microbes, species-specific primers were designedand screened. The results provide an important technology for investigation into thedynamic issues of the early-stage fouling microbes.This research was mainly composed of three parts: Analysis andidentification16SrDNA of in early-stage fouling microbes; Biochemical andphysiological research on marine early-stage fouling microbe species;Species-specific primer design and selection of early-stage fouling microbesConclusions obtained in this research are as follows:(1) By applying16SrDNA sequence analytic method, nineteen dominanttypes of marine fouling microbes at Xiaoshi Island area in summer were obtained, ofwhich5types were the majority. They were proteobacterium, Rhodobacteraceae,Roseovarius sp., Flavobacteriaceae and Alteromonadaceae seperately, occupying13%,10%,10%,9%and8%of the total bacterial community accordingly.(2) The results of identification early-stage fouling microbes obtained by pureculture are as follows: The physiological and biochemical analysis results indicatthat sixteen strains were the capsulated bacteria, of which twelve strains wereGram-negative bacteria, four strains the Gram positive bacteria, among which14strains were characterized as alteromonadaceae, proteobacterium, psychrobacterand marinomonas sp. accordingly. Of all14strains, eight strains have a similarityof99-100%compared to Alteromonadaceae, three have a similarity toproteobacterium, one has a similarity ratio up to99%compared with Psychrobacter,and still one has a similarity of99%to that of Marinomonas sp.(3) Through16SrDNA sequence analysis method and conventional pure culturemethod obtained dominant species in Xiaoshi Island area in summer.Compared thetwo methods, it was found that19strains from16SrDNA sequence analysis methodalready covered4strains obtained from pure culture method. So it was concluded that the major marine microbial species in the Xiaoshi Island of Weihai wereproteobacterium, Rhodobacteraceae, Roseovarius sp., Flavobacteriaceae andAlteromonadaceae. The16SrDNA sequence analysis method demonstrated itfeasible to analyze those marine bacteria which is difficult to separate only by purecultivation.(4) Two pairs of species-specific PCR primers were screened, it isdemonstrated that they can be used to successfully amplify target sequences todifferentiate Psychrobacter and Marinomonas sp.from marine microbes speciesscreened in Xiaoshi Island area in summer.