Abstract:Abstract:Microbial endocrinology is a crossdisciplinary field representing the intersection of microbiology with mammalian endocrinology and neurophysiology． In this review，effects of catecholamine on bacteria were used as an example to demonstrate the interactions between microbes and neuroendocrine hormones．Catecholamine modulates bacterial infectivity by stimulation of bacteria growth and augmentation of host tissue attachment and invasion．Moreover，the bacterial adrenergic receptors recognized by catecholamine and its relationship with quorum sensing signals were also addressed．This review will be helpful for understanding the interactions between microorganism and host as well as health breeding and food safety in animal industries．

Abstract:Abstract:The state of “viable but non-culturable”(VBNC) is a survival strategy adopted by microorganisms when exposed to environmental stress．With the increasingly serious problem of xenobiotics pollution，enhanced microbial processes that exploit the potential of microbes to remediate polychlorinated biphenyl-contaminated environments have been developed．Microorganisms represent a significant advance with respect to the transformation and degradation of polychlorinated biphenyls in the environment． It is of great importance to study the potential function of VBNC bacteria in polluted environment．In this paper，current research status of VBNC bacteria is summarized，and resuscitation of VBNC bacteria to potentially stimulate microbial degradation of pollutants is discussed in detail．Furthermore，we put forward a novel approach to explore the potential of VBNC bacteria for polychlorinated biphenyls degradation using resuscitation promoting factor (Rpf) of Micrococcus luteus．The novel efficient method is helpful for excavating and obtaining highly desirable polychlorinated biphenyls degrading microorganisms． Moreover，the prospect of VBNC bacteria to other environmental remediation fields，such as flocculation and nitrification deodorant，is addressed.

Abstract:Abstract:［Objective］In order to investigate the composition and diversity of prokaryotes in marine surface sediment from site XSCS13 at northern South China Sea.［Method］We extracted environment total DNA directly from the sediment and amplified 16S rRNA genes from the total DNA，thereby constructing 16S rRNA clone libraries of both archaea and bacteria. Then we selected positive clones randomly from the library and identified them by the method of restriction fragment length polymorphism (RFLP). After that，the unique RFLP pattern corresponded sequences were sequenced，BLAST and then constructed into a phylogenetic tree．［Results］ Most of the clones were sequences from uncultured microbes. For the Archaea part: the community was mainly comprised of 3 phyla: Crenarchaeota，Thaumarchaeota and Euryarchaeota，among which Crenarchaeota was the dominant class with the percentage of 71% ，while Euryarchaeota took the least with only 3 clones．The main group of Crenarchaeota was Marine Group Ⅰ，taking 61% of all. For the bacteria part，all together 9 phyla were included: 32.6% of Proteobacteria，3% of Verrucomicrobia，5.2% of Bacteroidete，4.44% of Acidobacteria，6% of Chloroflexi，3.7% of Firmicute，5.2% of Planctomycete，11.1% of Gemmatimonadete and 4.44% of Actinobacteria. Unlike archaea，there's no overwhelming preponderant phylum in bacterium，each phylum was relatively distributed in balanced proportions．Proteobacteria had 3 classes involved: α-Proteobacteri，γ-Proteobacteria and δ-Proteobacteria，and γ-Proteobacteria stood out with 54.5% of proportion．In addition，over half of all the clones were related to reduction reaction of sulfate and generation of methane． ［Conclusion］The research indicated that prokaryote diversity in marine surface sediment from site XSCS13 at northern South China Sea was quite plentiful，and a great mass of abundant microbial resources still remained unknown．Furthermore，the community structure of archaea and bacteria showed that the sampling site may be in a cold spring area with abundant methane.

Abstract:Abstract:［Objective］ To investigate the characteristics of classⅠintegrons among clinical multidrug-resistant Pseudomonas aeruginosa isolates，and to analyze the association between class Ⅰ integrons and multidrug resistance.［Methods］Pseudomonas aeruginosa strains were isolated from clinical samples，and the multidrug-resistant ones were picked out from these isolates．PCR assays were used to detect the variable regions of class Ⅰ integrons，and the resulting products were then digested with restriction enzyme Sau 3AⅠand sequenced． SPSS19. 0 software was used for statistical analysis.［Results］ Class Ⅰ integrons were detected in 27.3% of the clinical multidrug-resistant Pseudomonas aeruginosa isolates．Amplification of variable regions of class Ⅰ integrons revealed three different gene cassette arrays (1500，2300 and 4000 bp)，two of them were also found in other bacteria．These gene cassette arrays encoded aminoglycoside-modifying enzymes (aadA，aadB，aac(6')Ⅱ and aadA13)，β-lactamases (blaCARB8 and oxa10) and chloromycetin efflux pump (cmlA8). Analysis results suggested closely relationship between class Ⅰ integrons and aminoglycoside resistance.［Conclusion］We revealed three different gene cassette arrays in the clinical isolates of the multidrug-resistant Pseudomonas aeruginosa，and gene cassette array aadB-aac(6')Ⅱ-blaCARB8 was the most prevalent．The three gene cassette arrays all contained the aminoglycoside-resistant genes.

Abstract:Abstract:［Objective］Purpose of this work was to screen promoters which were induced by dissolved oxygen in Corynebacterium crenatum SYPA5-5． ［Methods］ Based on the genomic information of Corynebacterium，we identified target proteins which were the highest expression level of one copy of 27 known proteins from our earlier study．Based on the upstream sequence of the coding gene sequences，we amplified two promoters，named P-tkt and P-fum． The tac promoter of recombinant pDXW-8-cat and pDXW-8-gfp were replaced by the new promoters，and the recombinant plasmids were transformed into E.coli JM109 and C.crenatum SYPA5-5 respectively．At different dissolved oxygen level，we compared the function of promoters by the expression of chloramphenicol acetyltransferase (CAT) and green fluorescent protein (GFP).［Results］ Results show that we identified 2 target proteins，which were transketolase and fumarate hydratase．At the high dissolved oxygen level the CAT activity of E.coli JM109/pDXW-Ptkt-cat and C.crenatum SYPA5-5/pDXW-Ptkt-cat were 3.032 U/mg and 1.987 U/mg，which were 2.8-fold and 3.2-fold than that in the low dissolved oxygen，respectively． We got similar results by using a 5-L fermentor.［Conclusion］The P-tkt promoter induced by high dissolved oxygen was suitable for fermentation to produce amino acids and beneficial to improve ability of synthesis and metabolism of amino acids at high dissolved oxygen level.

Abstract:Abstract:［Objective］Phage infection could seriously influence cell growth and metabolism in the fermentation of 1，3-propanediol from glycerol by Klebsiella pneumoniae．Isolation of the Klebsiella pneumoniae phage and research on its physiological characteristics would be of great significance.［Methods］ A K．pneumoniae phage was isolated by using Adams double plate method from the infected fermentation broth of 1，3-propanediol. After the electron microscope observation，the genome of the phage was extracted and its size was identified with restriction enzyme analysis. The physiological characteristics of the phage were also tested，such as the optimal multiplicity of infection，the one-step growth curves and the sensitivity to temperature，pH，UV light and chloroform. And the phage infected fermentation was carried out and compared with normal fermentation.［Results］ The phage had an isometric polyhedral head (about 60 nm－70 nm in diameter) and a long noncontractile tail (about 160nm long)．The nucleic acid could be cut off by dsDNA restriction enzyme EcoRⅠor HindⅢ and its complete size was about 42 kb．It was sensitive to high temperature and UV light，insensitive to chloroform．The optimal multiplicity of infection for the phage was 1，the latent phase and rise phase were both 50min，and the burst size was 343．Compared with the normal fed-batch fermentation of 1，3-propanediol， the phage infected fermentation indicated that cell growth was delayed about 8 h and metabolic flow was changed to organic acid (e. g. lactic acid) pathway．［Conclusion］The phage was a non-envelop long-tailed phage，and could change the metabolism of the 1，3-propanediol fermentation from glycerol by Klebsiella pneumoniae． This work would be helpful for prevention and controlling of phage infection during the 1，3-propanediol fermentation．

Abstract:Abstract:［Objective］ We studied the antibacterial activities and mechanism of a new peptide P7，according to the structure-activity relationships of cell-penetrating peptide and antimicrobial peptide.［Methods］ The antimicrobial activities and cytotoxicity of P7 were examined using the microdilution and hemolysis analysis．The effects of P7 on the outer and plasma membrane permeability，membrane integrity and morphology of E.coli cells were analyzed by the membrane fluorescent probe，ow cytometry and scanning electron microscopy. Localization of the P7 onto the E.coli cells was determined by using a confocal laser-scanning microscopy． The DNA binding activity of P7 was evaluated by electrophoretic mobility shift assay.［Results］P7 possessed stronger antimicrobial activity than ppTG20. The inhibitory concentration was in the range from 4 to 32 μmol /L where P7 shown low hemolysis. P7 could increase the outer and plasma membrane permeability，induce the cell membrane integrity loss and cells structure damage. P7 penetrated the cell membrane，accumulated inside the cytoplasm and interacted with DNA. ［Conclusion］P7 exerted its antibacterial activity by increasing cell membrane permeability，penetrated the cell membrane and binding to DNA.

Abstract:Abstract:［Objective］ We isolated Pseudomonas strains from soil samples collected from Changsha，Hunan province，catalogued them and studied the antimicrobial and antitumor activity.［Methods］We isolated Pseudomonas strains from soil samples through Galleria bait method，identified and catalogued the isolated strains according to morphological observation，physiological and biochemical characteristics and the homologous analysis of 16S rRNA sequences of nucleotides．Antimicrobial，antagonistic fungi and anti-tumor activities were studied by diffusion plate assay，dual-culture assay and cytotoxicity test respectively.［Results］We isolated 5 Pseudomonas strains from vegetable field and forest land of Changsha suburb，catalogued and named them as Pseudomonas protegens CY01，Pseudomonas chlororaphis CY02，Pseudomonas oryzihabitans CY04，Pseudomonas sp. CY05 and Pseudomonas putida CY06 respectively. P. protegens CY01 and P. chlororaphis CY02 have antibacterial activity against Bacillus subtillis and Staphylococcus aureus. P. chlororaphis CY02 has antagonistic activity against Pyricularia oryzae and anti-tumor cell activity against mouse melanoma B16 cells.［Conclusion］The isolated P. chlororaphis CY02 has a significant effect on pathogenic bacteria，Pyricularia oryzae and tumor cells.

Abstract:Abstract:［Objective］The ClfA adhesin of Staphylococcus aureus is an excellent vaccine candidate antigen． CD4 + T cells play central roles during immune responses，but their functional contributions to Staphylococcus aureus in fection have yet to be evaluated． ［Methods］By using the SYFPEITHI prediction algorithm，we identified and characterized four Th epitopes within the ClfA adhesin.［Results］Peptide C335 was I-Ed restricted Th1 type epitopes; peptides C214，C286，and C436 were I-Ad restricted Th2 type epitope.［Conclusion］ The identification of these epitopes is important to evaluate and optimize the vaccine-primed protection against Staphylococcus aureus infection.

Abstract:Abstract:［Objective］Bartonella vinsonii subsp．berkhoffii is a fastidious haemotropic Gram-negative bacterium that has been identified as an emerging causative agent for zoonotic diseases of human and dogs．This study aimed to develop a TaqMan-MGB probe based，highly sensitive and species-specific fluorescence quantitative PCR assay for rapid detection of B．vinsonii subsp． berkhoffii．［Methods］The species-specific primersand probe set for B．vinsonii subsp．berkhoffii were designed．The annealing temperature，final concentration of the TaqMan-MGB probe and primers were optimized．Specificity，sensitivity and reproducibility of the PCR system were assessed．The standard curve was made using 10 × dilution series of the plasmid standard to analyze stability and PCR efficiency．［Results］ The real-time PCR with TaqMan-MGB assay was highly specific and sensitive for the detection of B．vinsonii subsp．berkhoffii．TaqMan-MGB probe-based fluorescence quantitative PCR did not show cross reactivity with the other Bartonella species，non-Bartonella bacteria and dogs and human．The detection limit of the TaqMan-MGB assay for the detection of B．vinsoniisub sp．berkhoffii was 11 copiesof the plasmid DNA per PCR reaction．The coefficient of variation CV% from the intra- and intergroupwas in the range of 0.12%－0.70% and 0.14%－0.55% which were acceptable．The correlation coefficient and E-value of the standard curve were 1.0 and 104.7%，which reflected a very good linearity and high efficiency．［Conclusion］ The TaqMan MGB-based probe fluorescence quantitative PCR assay was a reliable，species-specific，sensitive and useful tool for rapid detection of B.vinsonii subsp． berkhoffii.

Abstract:Abstract:［Objective］ The present study was to fully evaluate the intestinal bacterial community of Periplaneta americana，an important model to study insects.［Methods］We investigated the bacterial community of P. americanagut by culture-independent methods，involving constructing the 16S rRNA gene library and microbial diversity analysis.［Results］The phylotypes were affiliated with Proteobacteria(66.4%)，Bacteroidetes (17.8%)，Firmicutes (14.5%)，Fusobacteria (0.6%) and unclassified bacteria (0.6%)．Phylogenetic analysis shows that 15% of the sequences clustered with that from a closely related omnivorous cockroach;and 59% clustered with that from more distantly related animals，including omnivorous，herbivorous，and carnivorous animals，which differ greatly in feeding habits． Moreover，18% of the clones showed high sequence identity with potential pathogens closely related to human diseases，which also reinforces the concept of the cockroach as a carrier of pathogens．［Conclusion］Due to their habits of feeding on a variety of foodstuffs，omnivorous cockroaches harbor a large and diverse microbial community in the gut． The host phylogeny and dietary habits might be critical for the intestinal bacterial community composition of cockroaches.

Abstract:Abstract:［Objective］To search for structurally novel and biologically active compounds from the secondary metabolites of gorgonian-derived actinomyces.［Methods］ Strains of actinomyces with antimicrobial activities were screened by biological methods. Then，those active strains were cultured under different conditions to obtain crude extracts. Subsequently，the chemical diversities of the extracts were investigated by reverse phase high-performance liquid chromatography (RP-HPLC) and thin layer chromatography (TLC)，while the bioactivities of the extracts were evaluated by antifoulingand antibacterial assays. A target strain Streptomyces sp. SCSGAA0009 was selected to further study by large-scale fermentationon modified ISP2 medium. The compounds were isolated by column chromatography over silica gel，Sephadex LH-20，and semipreparative HPLC，and their structures were determinated by spectroscopic analysis.［Results］The target strain was identified as actinomyces，Streptomyces sp. SCSGAA0009. A new alkaloid N-(2-(1Hindol-3-yl) ethyl) propionamide (1) and a known compound phenazine-1-carboxylic acid (2) were purified from the culture broth. Compound 2 showed moderate antibacterial activity against Escherichia coli and Pseudoaltermonas piscida，and exhibited strong antilarval settlement activity towards Bugula neritina larva.［Conclusion］ Bioactive and new compounds can be achieved from the secondary metabolites of gorgonian-derived microorganisms from South China Sea.

Abstract:Abstract:［Objective］To determine blue-light induced expression of S-adenosyl-L-homocysteine hydrolase-like (sahhl) gene in fungus Mucor amphibiorum RCS1.［Methods］In the random process of PCR，a sequence of 555 bp was obtained from M. amphibiorum RCS1. The 555 bp sequence was labeled with digoxin to prepare the probe for northern hybridization. By northern hybridization，the transcription of sahhl gene was analyzed in M. amphibiorum RCS1 mycelia culture process from darkness to blue light to darkness. Simultaneously real-time PCR method was used to the sahhl gene expression analysis. ［Results］Compared with the sequence of sahh gene from Homo sapiens，Mus musculus and some fungi species，a high homology of the 555 bp sequence was confirmed． Therefore，the preliminary confirmation has supported that the 555 bp sequence should be sahhl gene from M． amphibiorum RCS1． Under the dark pre-culture in 24 h，a large amounts of transcript of sahhl gene in the mycelia can be detected by northern hybridization and real-time PCR in the condition of 24 h blue light． But a large amounts of transcript of sahhl gene were not found in other detection for the dark pre-culture of 48 h，even though M. amphibiorum RCS1 mycelia were induced by blue light. ［Conclusion］Blue light can induce the expression of sahhl gene in the vigorous growth of M. amphibiorum RCS1 mycelia.