Acinetobacter are a major concern because of their rapid development of resistance to a wide range of antimicrobials, and rapid profundity in transformation, surviving desiccation and persistinge in the environment for a very long time. The organisms are associated with bacteraemia, pulmonary infections, meningitis, diarrhea and notorious nosocomial infections with mortality rates of 20 to 60%. Transmission is via person-to-person contact, water and food contamination, and contaminated hospital equipment. The increasing virulence and rapid development of multidrug resistance by these organisms highlights the need to search for alternatives for chemotherapy. A poor understanding of the organisms and dearth of information about their occurrence especially in developing countries informed the need for this review paper.

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The seasonal change of bacterial community structure in biofilms on the surface of reed (Phragmites australis) was investigated for about three years (from 2005 June to 2008 March) in Lake Biwa by comparing it with that in surrounding lake water. The community structure in biofilms was different from that in the lake water throughout the seasons and years. The community structure in lake water was similar in the same seasons of different years, corresponding to similar environmental factors (i.e., temperature, dissolved oxygen, and light intensity) and nutrient ion concentrations at the same season. However, the community structure in the biofilms was not similar in the same season of different years. This seems to be due to the formation of new biofilms on sprouted reeds in every early summer and the high nutrient concentrations and bacterial density in subsequently formed biofilms. Although the community structure in the biofilms changed along with the seasonal change, the bacteria belonging to Bacillus and Paenibacillus were detected in any season. This study revealed the possibility that the bacterial community structure in the initial stage of the biofilm formation govern the subsequent seasonal change of the community structure in biofilms.

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The cyanobacterium Synechococcus leopoliensis CCAP1405/1 does not grow on common solid media made of agar, agarose HT, noble agar, gelrite and gelatin, although it grows in liquid media with the same components. The inoculation of S. leopoliensis CCAP1405/1 at a high initial cell density allowed it to grow on the agar media, and co-inoculation with one of the heterotrophic bacterial strains belonging to a wide range of phylogeny, showed the same effects even at a low initial cell density of S. leopoliensis CCAP1405/1. The addition of thiosulfate and high concentrations of vitamin B₁₂, biotin and thiamine also supported growth on solid media, but catalase had no effect. On inorganic solid media, the autotrophic cyanobacterial strain supported the growth of heterotrophic bacteria, suggesting mutual interaction.

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The relationship between the colonization of Lotus japonicus by plant growth-promoting fungi (PGPF) and biosynthesis of the isoflavonoid phytoalexin vestitol, a major defensive response of leguminous plants, was analyzed. When PGPF including Trichoderma koningi, Fusarium equiseti, and Penicillium simplicissimum were inoculated onto L. japonicus roots, only T. koningi colonized the roots long-term and increased plant dry weight (126%). Microscopic observations of transverse sections of roots colonized by T. koningi demonstrated intercellular hyphal growth and the formation of yeast-like cells. The induction of plant defenses by fungal infections was examined by Northern analysis of genes involved in vestitol biosynthesis and HPLC of vestitol production in L. japonicus. Inoculation with symbiotic Mesorhizobium loti did not induce any accumulation of the transcripts. T. koningi immediately suppressed transcript levels to those induced by M. loti. The vestitol transuded from roots by T. koningi was detected at a level equivalent to that transuded by M. loti. Other PGPF and Calonectoria ilicola pathogenic to soybean but not to L. japonicus, stimulated continuous expression of genes and exudation of vestitol. These PGPF resembled mycorrhizal fungi in the establishment of symbiotic associations rather than fungal parasites.

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Fluoroquinolone antibiotics (FQs) have been used worldwide for chemotherapy, animal husbandry, and aquaculture, and the occurrence of FQ-resistant (FQs^r) bacteria in natural environments has been reported. Plasmid-mediated transferable quinolone resistance (PMQR) genes are suspected to originate from the chromosomes of water-dwelling bacteria. However, the occurrence of and the potential reservoir of FQs^r bacteria and PMQR genes in aquatic environments have not been elucidated. In this study, we detected FQs^r bacteria and PMQR genes in aquatic environments in Thailand and Vietnam, and measured FQ contamination. Levels of contamination were greater Thailand (avg. 5130, max 46100 ng L^-1) than in Vietnam (avg. 235, max 1130 ng L^-1); however, the occurrence of FQs^r bacteria was higher in Vietnam (~15%) than in Thailand (~7.0%), suggesting that contamination by FQs is not directly linked to the development of FQs^r bacteria. Diverse taxonomic groups of FQs^r-bacteria were identified, and one of the PMQR genes, qnrB, was detected from bacteria of environmental origin, not enteric bacteria. This suggests that the environmental bacteria are a potential reservoir of antibiotic resistance determinants even at un-contaminated sites.

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N-Acylhomoserine lactones (AHLs) are used as quorum-sensing signaling molecules by many Gram-negative bacteria. Here, 413 bacterial strains were obtained from the roots of potato plants and screened for AHL-degrading bacteria using Chromobacterium violaceum reporter strains. Sixty one isolates degraded N-hexanoyl-L-homoserine lactone (C6-HSL) within 24 h. Based on their 16S rRNA gene sequences, most of these isolates were assigned to the genus Chryseobacterium and divided into eight groups. Most of the strains degraded N-decanoyl-L-homoserine lactone (C10-HSL) and were active against 3-oxo-substituted AHLs. Groups V and VI were more effective at inactivating AHLs. One group V strain, StRB402, showed putative AHL-lactonase activity. This is the first report of AHL-degrading Chryseobacterium strains.

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Microbial community composition in a pilot-scale microfiltration plant for drinking water treatment was investigated using high-throughput pyrosequencing technology. Sequences of 16S rRNA gene fragments were recovered from raw water, membrane tank particulate matter, and membrane biofilm, and used for taxonomic assignments, estimations of diversity, and the identification of potential pathogens. Greater bacterial diversity was observed in each sample (1,133-1,731 operational taxonomic units) than studies using conventional methods, primarily due to the large number (8,164-22,275) of sequences available for analysis and the identification of rare species. Betaproteobacteria predominated in the raw water (61.1%), while Alphaproteobacteria were predominant in the membrane tank particulate matter (42.4%) and membrane biofilm (32.8%). The bacterial community structure clearly differed for each sample at both the genus and species levels, suggesting that different environmental and growth conditions were generated during membrane filtration. Moreover, signatures of potential pathogens including Legionella, Pseudomonas, Aeromonas, and Chromobacterium were identified, and the proportions of Legionella and Chromobacterium were elevated in the membrane tank particulate matter, suggesting a potential threat to drinking water treated by membrane filtration.

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Rhizobia establish symbiosis with legumes. Bacteroids in indeterminate nodules of Inverted Repeat Lacking Clade (IRLC) legumes undergo terminal differentiation caused by Nodule-specific Cysteine-Rich peptides (NCRs). Microscopic observations of bacteroids and the detection of NCRs in indeterminate nodules of the non-IRLC legume Leucaena glauca were performed. A portion of the bacteroids showed moderate cell elongation, loss of membrane integrity, and multiple nucleoids. The symbiosome contained multiple bacteroids and NCR-like peptides were not detectable. These results indicate that bacteroid differentiation in L. glauca is different from that in IRLC legumes although both hosts form indeterminate nodules.

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Pseudomonas aeruginosa, an opportunistic pathogen, is known to mainly use N-acylhomoserine lactones (AHLs) as autoinducers. Recent progress in this field demonstrated that not only AHLs, but also their degradation products, tetramic acids, may have some biological activities. The present study examined the roles of Pseudomonas autoinducers and tetramic acids in bacterial survival and behavior in ecological niches. P. aeruginosa autoinducers and the tetramic acid derivatives were chemically synthesized, and the structure-activity correlation was investigated. Some tetramic acids derived from AHLs caused a significant reduction in the viability of P. aeruginosa in a concentration dependent manner (30-300 μM). The smaller the inoculum of bacteria, the stronger the bactericidal activity that was observed. The data from tetramic acid derivatives indicated the keto-enol structure of tetramic acid to be critical for the antibacterial activity. Exogenous tetramic acid did not induce significant changes in the formation of biofilm or production of exoproducts such as pyocyanin and elastase. Tetramic acid and disinfectants acted synergistically to kill P. aeruginosa. These results suggest the AHL-degradation product tetramic acid to be useful for bacterial control.

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Signature-tagged mutagenesis was applied to Mesorhizobium loti, a nitrogen-fixing root-nodule symbiont of the leguminous plant Lotus japonicus. We arranged 1,887 non-redundant mutant strains of M. loti into 75 sets, each consisting of 24 to 26 strains with a different tag in each strain. These sets were each inoculated en masse onto L. japonicus plants. Comparative analysis of total DNA extracted from inoculants and resulting nodules based on quantitative PCR led to the selection of 69 strains as being reduced in relative abundance during nodulation. Plant assays conducted with individual strains confirmed that 3 were defective in nodulation (Nod^-) and that 10 were Nod⁺ but defective in nitrogen fixation (Fix^-); in each case, the symbiosis deficiency could be attributed to the transposon insertion carried by that strain. Although the remaining 56 strains were Fix⁺, 33 of them showed significantly reduced competitiveness during nodulation. Among the mutants we identified are known genes that are diverse in predicted function as well as some genes of unknown function, which demonstrates the validity of this screening procedure for functional genomics in rhizobia.

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The aims of this study were to isolate Aurantimonas and Methylobacterium strains that responded to soybean nodulation phenotypes and nitrogen fertilization rates in a previous culture-independent analysis (Ikeda et al. ISME J. 4:315-326, 2010). Two strategies were adopted for isolation from enriched bacterial cells prepared from stems of field-grown, hypernodulated soybeans: PCR-assisted isolation for Aurantimonas and selective cultivation for Methylobacterium. Thirteen of 768 isolates cultivated on Nutrient Agar medium were identified as Aurantimonas by colony PCR specific for Aurantimonas and 16S rRNA gene sequencing. Meanwhile, among 187 isolates on methanol-containing agar media, 126 were identified by 16S rRNA gene sequences as Methylobacterium. A clustering analysis (>99% identity) of the 16S rRNA gene sequences for the combined datasets of the present and previous studies revealed 4 and 8 operational taxonomic units (OTUs) for Aurantimonas and Methylobacterium, respectively, and showed the successful isolation of target bacteria for these two groups. ERIC- and BOX-PCR showed the genomic uniformity of the target isolates. In addition, phylogenetic analyses of Aurantimonas revealed a phyllosphere-specific cluster in the genus. The isolates obtained in the present study will be useful for revealing unknown legume-microbe interactions in relation to the autoregulation of nodulation.

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PFGE has potential applications in the source tracking of fecal pollution in aquatic environments. We tried to distinguish the genotypes of Enterococcus faecium collected from fecal-contaminated water using PFGE. Well identified 115 strains of E. faecium were classified into 25 PFGE patterns, and characteristics distinctive to each genotype were recognized. Analysis of the characteristics of genotypes using PFGE can be used to track source of fecal pollution.

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Two-dimensional gel electrophoresis (2-DGE) mapping of genomic DNA and complementary DNA (cDNA) amplicons was attempted to analyze total and active bacterial populations within soil and activated sludge samples. Distinct differences in the number and species of bacterial populations and those that were metabolically active at the time of sampling were visually observed especially for the soil community. Statistical analyses and sequencing based on the 2-DGE data further revealed the relationships between total and active bacterial populations within each community. This high-resolution technique would be useful for obtaining a better understanding of bacterial population structures in the environment.

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