The Prevention And Control Of Some Common Health-care Associated Bacteria Using Bacteriophage

With the increasing of antibiotic resistant bacteria, especially the emergence anddissemination of “superbug bacteria”, people worries about that human may enter intothe period of post-antibiotic era when no antibiotic for use. Bacteriophage wasdiscovered earlier than antibiotics and then they had been used to control infectiousdisease caused by bacteria once they were discovered. However, phage therapy werenearly ignored with the industrial production and application of antibiotics. Recently,phage therapy have been refocused by researchers.Our aims are based in Shanghai, firstly, we isolate some hospital-care associatedbacteria （P. aeruginosa, S. aureus, K.pneumonia and Enterococcus）. Secondly, thesamples such as soils, feces, and sewage water et al may contain bacteriophage werecollected from13provinces or directly governed city region. The bacteriophages wereisolated from these samples. Finally we establish a pool of bacteriophage thatinfecting some common hospital associated bacteria, including22phages of P.aeruginosa,2phages of S. aureus,1phage of K. pneumonia, and2phages ofEnterococcus. It can provide foundation for using bacteriophage control infectiousdiseases caused by bacteria. We identified a new Staphylococcus phage whichbelongs to unclassified Twortlikevirus, and named Staphylococcus phage JD007;besides we also identified a new Klebsiella phage which belongs to unclassifiedMyoviridae, and we named Klebsiella phage JD001; furthermore, the Pseudomonasphage JD024we discovered did not contain anti-CRISPR genes of JBD5₀35andJBD5₀36, it can provide the clues for the evolution of Pseudomonas phages.The complete genome sequence and characterization of Klebsiella phage JD001,Staphylococcus phage JD007and Pseudomonas phage JD024were studied. Ourresults show that these three bacteriophage all belong to lytic phage, there are not anyvirulent genes or antibiotic resistant genes were found in the genome of these phages.All of them can keep good activity at37℃or at pH7for1h. The Staphylococcus phage JD007can kill the94％S. aureus （including methicillin resistant S.aureus）.The results showed its potential use for phage therapy.We found two endolysins named LysKP2and LysKP1that containinglysozyme-like activity domain at2ORFs and59ORFs in the whole genome sequenceof phage JD001predicted by the bioinformatics. LysKP1can kill the bacteria whenexpressing in it. Further study show that LysKP2can destroy the cell wall of K.pneumonia. It have lysozyme-like activity, the optimal temperature of theLysozyme-like activity of LysKP2is about30℃, lysozyme-like activity of lysozymekeeps high between the pH7and pH10. EDTA, Cu2+, Mg2+and Triton X-100canenhance its activity, while SDS can inhibit its activity. Our study show that whenLysKP1were induced by IPTG in E.coli BL21, it can kill the E.coli BL21. Our canprovide the foundation for acknowledge the.mechanism of JD001killing its host.In summary, the study established a pool of bacteriophage infecting somecommon hospital acquired bacteria （P.aeruginosa, S. aureus, K. pneumonia andEnterococcus）. Klebsiella phage JD001, S. aureus phage JD007and Pseudomonasphage JD024can potentially use for phage therapy according their characterizationsand complete genome. The LysKP2of Klebsiella phage JD001can destroy thestructure of Klebsiella pneumonia from outside, and it has muralytic activity, itprovide foundation for acknowledge of mechanism of Klebsiella phage JD001killingKlebsiella pneumonia. Our study can provide the foundation for the development ofnew antimicrobial agents used for the prevention or control of bacteria in the nearfuture. Background: An increasing number of studies have implicated the microbiomein certain diseases, especially chronic diseases. In this study, the bacterialcommunities in the sputum of pulmonary tuberculosis patients were explored. TotalDNA was extracted from sputum samples from31pulmonary tuberculosis patientsand respiratory secretions of24healthy participants. The16S rRNA V3hyper-variable regions were amplified using bar-coded primers and pyro-sequencedusing Roche454FLX. Results: The results showed that the microbiota in the sputumof pulmonary tuberculosis patients were more diverse than those of healthyparticipants （p<0.05）. The sequences were classified into24phyla, all of which werefound in pulmonary tuberculosis patients and17of which were found in healthyparticipants. Furthermore, many foreign bacteria, such as Stenotrophomonas,Cupriavidus, Pseudomonas, Thermus, Sphingomonas, Methylobacterium,Diaphorobacter, Comamonas, and Mobilicoccus, were unique to pulmonarytuberculosis patients. Conclusions: This study concluded that the microbialcomposition of the respiratory tract of pulmonary tuberculosis patients is morecomplicated than that of healthy participants, and many foreign bacteria were found inthe sputum of pulmonary tuberculosis patients. The roles of these foreign bacteria inthe onset or development of pulmonary tuberculosis shoud be considered byclinicians.