Preliminary Research On Effect Of Nanomaterials On The Process Of Early-stage Marine Fomling Microbial Attachment

Marine fouling organisms, which can attach to ships and varieties of underwaterartificial facilities, have a negative impact on human economic activities. Theformation process of early marine microbial film is an important node to the study onattaching dynamics process of the entire marine fouling organisms. Therefore, howto interfere and inhibit the formation of biofilm is of great significance to preventbiofouling. Nanomaterials have the characteristics of surface effect and quantum sizeeffect, thereby affecting the structure and function of biological macromol eculessuch as proteins, nucleic acids, etc. Currently, a host of countries considernano-materials as a research focus into the marine antifouling paints aiming to solvethe problem of attaching marine fouling organismsThis topic first explores the experiment conditions of seaside artificial hangedplate, and then screens the anti-fouling effect of77kinds of carbon nanotubes andother10kinds of nanomaterials. In addition, gene cloning method is applied toidentify early microbial strains. Finally regular PCR and Q-PCR is employed toscreen the anti-fouling effect of13kinds of nanomaterials and combinations.We extract earlier microbial genomic DNA, amplify16SrDNA by regular PCR,ligate it to pMD19-T plasmid vector, transform into E coli strain and finallyconstruct early fouling microbial16SrDNA gene library. Positive clones are screenedby blue-white selection, then use restriction endonuclease enzyme Hindш and SacIdigestion. After sequencing, alignment and homology analysis, we compare theknown bacteria16SrDNA sequence homology with the NCBI website, finding16early microbial strains with the similarity up to99%. Based on whole genomesequences of some of those strains, species-specific primers have been designed.Hanging-plate experiment is to paint the steel surface with certainconcentrations of nanomaterials to observe the inhibitory effect on the foulingorganisms. Through a large scale screening of77kinds of carbon nanotubes andother10kinds of nanomaterials, we eventually filter out13kinds of nano-materialsand combinations which demonstrates apparent inhibition effect towards biofouling:they are Number3,41,51,76carbon nanotubes, carbon nanotubes combinationsNumber8-51,3-49,11-50,2-14,2-33,27-61,25-66,50-67, as well as Si-Nicombinations respectively,both of which with the concentration of2%.By detecting the antifouling effect of13nanomaterials combinations usingregular PCR, we find13nanomaterials and combinations all appeared apparentinhibition effect on fouling microbes attachment, especially towardsPseudoalteromonas atlantica, Marinomonas pontica, and Psychrobacter marincola. Using Q-PCR to screen Pseudoalteromonasissachenkonii strain we find that13nanometer materials demonstrated different inhibition effect on the microbialadherent function, According to the Ct value of Q-PCRreaction we can conclude thebest is No.76single-orientation carbon nanotubes.