ALBERT

Description: Quantitative real-time reverse transcription PCR (qRT-PCR) is a rapid, sensitive, and reliable technique for gene expression studies. The accuracy and reliability of qRT-PCR results depend on the stability of the reference genes used for gene normalization. Therefore, a systematic process of reference gene evaluation is needed. Ganoderma lucidum is a famous medicinal mushroom in East Asia. In the current study, 10 potential reference genes were selected from the G. lucidum genomic data. The sequences of these genes were manually curated, and primers were designed following strict criteria. The experiment was conducted using qRT-PCR, and the stability of each candidate gene was assessed using four commonly used statistical programs—geNorm, NormFinder, BestKeeper, and RefFinder. According to our results, PP2A was expressed at the most stable levels under different fermentation conditions, and RPL4 was the most stably expressed gene in different tissues. RPL4, PP2A, and β-tubulin are the most commonly recommended reference genes for normalizing gene expression in the entire sample set. The current study provides a foundation for the further use of qRT-PCR in G. lucidum gene analysis.

Description: The study aimed at optimization of DNA isolation from blood of representatives of four microbial groups causing sepsis, i.e., Gram negative: Escherichia coli , Gram positive: Staphylococcus aureus, yeast: Candida albicans , and filamentous fungus: Aspergillus fumigatus . Additionally, the five commercial kits for microbial DNA isolation from the blood were tested. The developed procedure of DNA isolation consisted of three consecutive steps, i.e., mechanical disruption, chemical lysis, and thermal lysis. Afterward, DNA was isolated from the previously prepared samples (erythrocyte lysis) with the use of five commercial kits for DNA isolation. They were compared paying heed to detection limit, concentration, DNA purity, and heme concentration in samples. The isolation of DNA without preliminary erythrocyte lysis resulted in far higher heme concentration than when lysis was applied. In the variant with erythrocyte lysis, two of the commercial kits were most effective in purifying the DNA extract from heme. Designed procedure allowed obtaining microbial DNA from all four groups of pathogens under study in the amount sufficient to conduct the rtPCR reaction, which aimed at detecting them in the blood.

Description: Culture-dependent evaluation of the bacteria was carried out on gastropods, such as Monodonta lineata, Gibbula umbilicalis, Nucella lapillus and Patella intermedia , and the environmental samples (biofilm and surrounding sea water) collected from six different locations of Northern Portugal coastal area to investigate the interactions between the microbes in the viscera of gastropods and in the environment. A total of 141 isolates and 39 operational taxonomic units were identified. Phylogenetic analysis based on the 16S rRNA gene showed that bacterial isolates are highly diverse and most of them were found in other marine environment. The observed bacterial diversity was distributed over five different classes (Gammaproteobacteria, Alphaproteobacteria, Flavobacteria, Bacilli and Actinobacteria) with the greatest number of 16S rRNA gene sequence derived from the Gammaproteobacteria (77 %). Vibrio is found to be the dominant one among the different bacterial species isolated. The results suggest that the microorganisms in the environment are maintained in the viscera of the gastropods which may have a key role in the metabolic functions.

Description: Two novel aerobic p - n -nonylphenol-degrading bacterial strains were isolated from seawater obtained from the coastal region of Ogasawara Islands, Japan. The 16S rRNA gene sequence analysis indicated that the strains are affiliated with the order Alteromonadales within the class Gammaproteobacteria . One isolate, strain KU41G2, is most closely related to Maricurvus nonylphenolicus (99.2 % similarity), and is tentatively identified as M. nonylphenolicus. The other isolate, strain KU41G T , is also most closely related to M. nonylphenolicus; however, the 16S rRNA gene sequence similarity was only 94.7 %. Cells of strain KU41G T are Gram-negative rods with a single polar flagellum. The predominant respiratory lipoquinone was ubiquinone-8, and the major cellular fatty acids were C 17:1 ω8c (24.2 %); C 15:0 iso 2-OH; and/or C 16:1 ω7c (16.3 %), C 15:0 (10.3 %), C 11:0 3-OH (9.5 %), C 9:0 3-OH (6.7 %), C 10:0 3-OH (6.4 %), and C 18:1 ω7c (5.5 %). The DNA G+C content was 53.3 mol%. On the basis of physiological, chemotaxonomic, and phylogenetic data, strain KU41G T is suggested to represent a novel species of a new genus, for which we propose the name Pseudomaricurvus alkylphenolicus gen. nov., sp. nov. The type strain of P. alkylphenolicus is KU41G T (=JCM 19135 T  = KCTC 32386 T ).

Description: MreB is a cytoskeletal protein, which is responsible for maintaining proper cellular morphology and is essential for cell survival. Likewise, penicillin-binding protein 5 (PBP5) helps in maintaining cell shape, though non-essential for survival. The contradicting feature of these two proteins paves the way for this study, wherein we attempt to draw a relation on the nature of distribution of MreB in PBP deletion mutants. The study revealed that the uniform MreB helices/patches were destabilized/disturbed at the zone of deformities of the PBP mutants, whereas the helical patterns were retained at the regions maintaining a rod shape. We interpret that MreB remains functional irrespective of its distribution being misguided by the aberrant shapes of PBP mutants.

Description: Lead is an important environmental pollutant. The role of vacuole, in Pb detoxification, was studied using a vacuolar protein sorting mutant strain ( vps16 Δ), belonging to class C mutants. Cells disrupted in VPS16 gene, did not display a detectable vacuolar-like structure. Based on the loss of cell proliferation capacity, it was found that cells from vps16 Δ mutant exhibited a hypersensitivity to Pb-induced toxicity, compared to wild type (WT) strain. The function of vacuolar H + -ATPase (V-ATPase), in Pb detoxification, was evaluated using mutants with structurally normal vacuoles but defective in subunits of catalytic ( vma1 Δ or vma2 Δ) or membrane domain ( vph1 Δ or vma3 Δ) of V-ATPase. All mutants tested, lacking a functional V-ATPase, displayed an increased susceptibility to Pb, comparatively to cells from WT strain. Modification of vacuolar morphology, in Pb-exposed cells, was visualized using a Vma2p-GFP strain. The treatment of yeast cells with Pb originated the fusion of the medium size vacuolar lobes into one enlarged vacuole. In conclusion, it was found that vacuole plays an important role in the detoxification of Pb in Saccharomyces cerevisiae ; in addition, a functional V-ATPase was required for Pb compartmentalization.

Description: Early studies on the coloured particles that fell as red rain over southern India identified them as unicellular eukaryotes such as members of the red algae or fungi; however, the results of the present investigation are not consistent with this designation. Using transmission electron microscopy, we have demonstrated significant differences in the ultrastructure when compared with representative species from these other groups. Most notably, the red rain cells show no evidence of typical eukaryotic internal structures such as mitochondria or endoplasmic reticulum. Furthermore, comparisons based on elemental composition using energy-dispersive X-ray analysis, as well as Raman spectral signatures demonstrate significant dissimilarities in their molecular composition. The identity and origins of the red rain cells remain an enigma; however, our findings are more consistent with an unidentified prokaryote, and thus suggest that previous attempts at their identification should be reappraised.

Description: This study provides a comprehensive picture of the C. neoformans/C. gattii molecular types most often associated with human cryptococcosis in Portugal and assesses the impact of C. gattii in these infections. One hundred and twenty-two clinical isolates, from distinct patients, were identified as C. neoformans and genotyped by URA5 -RFLP, with the molecular types VNI (45.5 %) and VNIII (30.9 %) being the most commonly found ones. The molecular types VNII (11.4 %) and VNIV (11.4 %) were less abundant. One patient was found to be infected with a VGII isolate. This patient exhibited unusual clinical symptoms of cryptococcosis, reinforcing the suspicion for the presence of a different genotypic pattern, as determined afterwards. This case was detected in 2007 and is the first report of a potential autochthonous C. gattii infection case in Portugal, as the patient revealed no historical record of travelling outside the country.

Description: Bacteriophage genes offer a potential resource for development of new antibiotics. Here, we identify at least six genes of Staphylococcus aureus phage Sb-1 that have bactericidal activity when expressed in Escherichia coli . Since the natural host is gram-positive, and E. coli is gram-negative, it is likely that a variety of quite different bacterial pathogens would be susceptible to each of these bactericidal activities, which therefore might serve as the basis for development of new wide-spectrum antibiotics. We show that two of these gene products target E. coli protein synthesis.

Description: The ESX-1 secretion system exports substrate proteins into host cells and is crucial for the pathogenesis of Mycobacterium tuberculosis . EspR is one of the characterized transcriptional regulators that modulates the ESX-1 system by binding the conserved EspR binding sites in the promoter of espA , the encoding gene of EspA, which is also a substrate protein of the ESX-1 system and is required for the ESX-1 activity. EspR is autoregulatory and conserved EspR binding sites are present upstream of espR . In this study, we showed that these EspR sites had varying affinities for EspR, with site B being the strongest one. Point mutations of the DNA sequence at site B abolished binding of EspR to oligonucleotides containing site B alone or with other sites, further suggesting that site B is a major binding site for EspR. Complementation studies showed that constructs containing espR , and the upstream intergenic region fully restored espR expression in a Δ espR mutant strain. Although recombinant strains with mutations at more than one EspR site showed minimal differences in espR expression, reduced expression of other EspR target genes was observed, suggesting that slight changes in EspR levels can have downstream regulatory effects. These findings contribute to our understanding of the regulation of the ESX-1 system.

Description: Artificial plasmid DNA transformation of Escherichia coli induced by calcium chloride is a routine technique in molecular biology and genetic engineering processes, but its mechanism has remained elusive. Because adenosine monophosphate (AMP) has been found to regulate natural transformation in Haemophilus influenza , we aimed to investigate the effects of AMP and its derivatives on E. coli transformation by treating competence with different concentrations of them. Analysis of the transformation efficiencies revealed that AMP inhibited the artificial plasmid DNA transformation of E. coli in a concentration- and time-dependent manner. Furthermore, we found that AMP had no effect on the expression of the transformed gene but that the intracellular AMP level of the competent cells rose after a 6 h treatment. These results suggested that the intracellular AMP level had an important role in E. coli transformation. And these have useful implications for the further investigation of the mechanism of E. coli transformation.

Description: Gcn5 is a well-established histone acetyltransferase involved in chromatin modification by catalyzing the acetylation of specific lysine residues within the N-terminal tails of the core histones. To assess the role of chromatin remodeling in the transcriptional response of cellulolytic Trichoderma reesei to the changes of environmental conditions, we identified the T. reesei ortholog of Saccharomyces cerevisiae Gcn5 by sequence alignment and functional analysis. Heterologous expression of TrGcn5 in S. cerevisiae gcn5 Δ strain restored the growth defect under nutrient limitation as well as stresses. In contrast, mutant TrGcn5 with site-directed changes of residues critical for Gcn5 histone acetyltransferase activity could not complement the growth defect. The T. reesei gcn5 Δ mutant strain displayed a strongly decreased growth rate and dramatic morphological changes including misshapen hyphal cells and abolished conidiation. Moreover, the induced expression of cellulase genes was severely impaired in the gcn5 Δ T. reesei with acetylation of K9 and K14 of histone H3 in the cellulase gene promoter dramatically affected in the absence of TrGcn5. The results indicate that TrGcn5 plays a critical role in filamentous growth, morphogenesis, and transcriptional activation of specific genes including cellulase encoding genes.

Description: An environmental freshwater bacterial isolate, DM104, appearing as Shigella -like colonies on selective agar plates was found to show strong and specific serological cross-reactivity with Shigella dysenteriae type 4. Biochemical identification according to the analytical profile index, molecular serotyping by restriction of the amplified O-antigen gene cluster ( rfb -RFLP), together with phylogenetic analysis of the 16S rRNA gene and multi-locus sequence analysis, identified the isolate as Escherichia albertii . rfb -RFLP of DM104, revealed a profile different from that of S. dysenteriae type 4. However, western blot analysis of extracted lipopolysaccharides demonstrated strong cross-reactivity with S. dysenteriae type 4 using specific monovalent antisera and a lipopolysaccharide gel banding profile similar to that of S. dysenteriae type 4. The observed O-antigen cross-reaction between an E. albertii isolate and S. dysenteriae extends our knowledge of the extent of O-antigen cross-reaction within the Escherichia/Shigella group of organisms, and offers the possibility of using DM104 and similar cross-reacting strains as shigellosis vaccine candidates.

Description: Most cases of fungal bloodstream infections (BIs) are attributed to Candida albicans ; however, non- Candida albicans Candida species have recently been identified as common pathogens. Although hemolytic factor is known to be putative virulence factor contributing to pathogenicity in Candida species, its production is poorly evaluated. The present study was undertaken to analyze the production of hemolytic factor by C. albicans (10), C. tropicalis (13), and C. parapsilosis (8) isolates associated with BIs. Data of hemolysis zones on plate assay revealed that the majority of C. albicans isolates produced mild hemolytic activity whereas the majority of C. tropicalis produced strong activity. None of the tested C. parapsilosis isolates exhibited hemolysis on plate assay. We also evaluated the hemolytic activity in the cell-free broth. There were no significant differences ( P  〉 0.05) in the secreted hemolytic activity among intra-species isolates. Different levels of secreted hemolytic factor were observed for Candida species, where C. tropicalis exhibited the highest production of hemolytic factor ( P  〈 0.05) followed by C. albicans and C. parapsilosis . Inhibition of hemolysis (up to 89.12 %) from culture supernatant, following incubation with the lectin Concanavalin A (Con A), was observed for all three Candida species. This finding suggests that the secreted hemolytic factor of C. tropicalis and C. parapsilosis may be a mannoprotein, similar to that described for C. albicans .

Description: Halophilic archaeal strain GX31 T was isolated from a marine solar saltern of China. The cells of the strain were rod-shaped and lysed in distilled water, stain Gram-negative and formed red-pigmented colonies. It was neutrophilic, and required at least 0.9 M NaCl and 0–1.0 M MgCl 2 for growth under the optimum growth temperature of 37 °C. The major polar lipids of the strain were phosphatidylglycerol (PG), PG phosphate methyl ester, PG sulphate, and two major glycolipids chromatographically identical to sulphated mannosyl glucosyl diether (S-DGD-1) and mannosyl glucosyl diether (DGD-1), respectively. Trace amounts of two unidentified lipids were also detected. On the basis of 16S rRNA gene sequence analysis, strain GX31 T was closely related to the members of Halobellus of the family Halobacteriaceae with similarities of 94.1–98.7 %. Strain GX31 T showed 89.8–95.4 % of the rpoB′ gene similarity to the members of Halobellus . The DNA G+C content of strain GX31 T was 66.8 mol%. Strain GX31 T showed low DNA–DNA relatedness with two most related members of the genus Halobellus . The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain GX31 T represent a novel species of the genus Halobellus , for which the name Halobellus litoreus sp. nov. is proposed. The type strain is GX31 T (=CGMCC 1.10387 T  = JCM 17118 T ).

Description: Streptomyces diastatochromogenes 1628, capable of producing toyocamycin (TM), has exhibited a potential biocontrol effect in inhibiting the development of phytopathogens in the agriculture field. In this study, an efficient transformation system was developed using the intergeneric conjugation. This was achieved by optimization of experimental parameters. Under optimal conditions, a maximal conjugation frequency of 4.1 × 10 −4 per recipient was obtained. In order to heterologously express the gene vgb encoding Vitreoscilla hemoglobin in S. diastatochromogenes 1628, we placed vgb under the control of the constitutive promoter Perm E * and constructed plasmid pIB139- vgb . This plasmid was integrated into the chromosome of S. diastatochromogenes 1628 using intergeneric conjugation established above. Finally, strain 1628-VHB-23 with the highest TM production was screened. Results indicated that expression of vgb gene had always significantly promoted the cell growth and TM production in S. diastatochromogenes 1628 under different dissolved oxygen conditions. In particular, under the limited aerobic condition, strain 1628-VHB-23 obtained 33.3 % more DCW and produced 210 % more TM in 7-l fermentor as compared with the wild-type strain.

Description: Many studies have demonstrated that the properties of enzymes expressed in eukaryotes can be affected by the position and extent of glycosylation on enzyme. In this study, two potential glycosylation sites (the 8th and the 58th asparagine) were identified and the effect of propeptide glycosylation on Rhizomucor miehei lipase (RML) expressed in Pichia pastoris was investigated. To better understand the effect of glycosylation on the activity of RML, three mutants (M1, generated by N8A; M2, generated by N58A; and M3, generated by N8A and N58A) were designed to generate deglycosylated enzymes. The results showed that deglycosylated RML exhibited a twofold higher activity compared to the wild type. However, it was also found that glycosylation on the propeptide was important for the removal of the propeptide by Kex2 protease and secretion of the enzyme. Thus, our study provided a further understanding into the role of glycosylation on enzyme function.

Description: Vancomycin-resistant Enterococci (VRE) is a serious concern for public health. Serious infections with VRE have very limited effective antimicrobial therapy, and alternative treatment approaches are highly desirable. One promising approach might be the photodynamic antimicrobial chemotherapy. In the present study, we investigated the photodynamic inactivation (PDI) of two VRE strains mediated by 5-aminolevulinic acid (5-ALA) and its derivative 5-ALA methyl ester (MAL). The photodynamic damages to bacteria on the level of genomic DNA, the leakage of cell components, and the changes of membrane structure were investigated. After treated with 10 mM 5-ALA and irradiated by the 633 ± 10 nm LED for 60 min, 5.37 and 5.22 log 10 reductions in bacterial survival were achieved for the clinical isolate of VRE and E. faecalis (ATCC 51299), respectively. After treated with 10 mM MAL and irradiated by the LED for 60 min, 5.02 and 4.91 log 10 reductions in bacterial survival were observed for the two VRE strains, respectively. In addition, the photocleavage on genomic DNA and the rapid release of intracellular biopolymers were detected in PDI-treated bacteria. The intensely denatured cytoplasm and the aggregated ribosomes were also found in PDI-treated bacteria by transmission electron microscopy. Although 5-ALA and MAL-mediated PDI could induce the photocleavage on genomic DNA, the PDI of the two VRE strains might be predominantly attributed to the envelope injury, the intracellular biopolymers leakage, and the cytoplasm denature.

Description: The biosynthesis of integric acid, a secondary metabolite of the wood-decay fungus Xylaria feejeensis strain 2FB-PPM08M, has been studied. Labeling experiments using [1- 13 C], [2- 13 C] and [1,2- 13 C 2 ] acetate and l -methionine (methyl- 13 C) were separately performed with fungal culture. The labeling patterns of these metabolites indicated the same origin, and determined that integric acid was formed through the condensation of a sesquiterpene and a polyketide. These experiments showed that side chain of compounds would be synthesized by the polyketide pathway, while the ring carbon indicated the biosynthesis of compounds via the mevalonate pathway.

Description: Two-component systems are important regulatory systems that allow bacteria to adjust to environmental conditions, and in some bacteria are used in pathogenesis. We identified a novel two-component system in Burkholderia cenocepacia , an opportunistic pathogen that causes pneumonia in cystic fibrosis (CF) patients. The putative operon encodes BceS, a sensor kinase, and BceR, a response regulator. Our studies indicated that the bceR mutant showed a statistically significant decrease in protease, swimming motility, and quorum sensing when compared to the wild-type, but there was no significant difference in phospholipase C activity, swarming, and biofilm formation. In addition, the mutant showed a statistically significant reduction in virulence compared to the wild-type using the alfalfa plant model. Examination of the Burkholderia cepacia complex (a group of organisms that are phenotypically similar, but genotypically distinct) revealed that this system is prevalent in B. ambifaria , B. multivorans , B. vietnamiensis and B. dolosa . Interestingly, all these organisms have been associated with CF patients. The collective results indicate that BceSR influences various activities important in Burkholderia physiology and possibly pathogenesis. This information could be important in the design of novel therapeutics for Burkholderia infections.

Description:    Fifty-five bacteriocinogenic lactic acid bacteria (LAB) isolated from seven different sources. Eight isolates were found to produce pediocin PA-1 like bacteriocin as detected by ped B gene PCR and dot-blot hybridization. The culture filtrate (CF) activity of these isolates exhibited strong antilisterial, antibacterial activity against tested food-borne pathogens and LAB. The identification and genetic diversity among the selected LAB was performed by conventional morphological and molecular tools like RFLP, RAPD, and 16S rDNA gene sequencing. The isolates were identified as, 1 each of Pediococcus acidilactici Cb1, Lactobacillus plantarum Acr2, and Streptococcus equinus AC1, 2 were of P. pentosaceus Cb4 and R38, and other 3 were Enterococcus faecium Acr4, BL1, V3. Partial characterization of the bacteriocins revealed that the peptide was heat-stable, active at acidic to alkaline pH, inactivated by proteolytic enzymes, and had molecular weight around 4.6 kDa and shared the properties of class IIa pediocin-family. The bacteriocin production at different temperatures, pH, and salt concentrations was studied to investigate the optimal condition for application of these isolates as a starter culture or as a biopreservative in either acidic or non-acidic foods. Content Type Journal Article Pages 1-5 DOI 10.1007/s00284-011-9963-8 Authors S. Manjulata Devi, Department of Food Microbiology, CFTRI, Mysore, India Prakash M. Halami, Department of Food Microbiology, CFTRI, Mysore, India Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The ability of the foodborne pathogen Listeria monocytogenes to develop biofilm in food-processing environment is a major concern for the food safety, because biofilms allow bacteria to better resist environmental stresses. PrfA is a key transcriptional activator that positively regulates most of the known listerial virulence gene expression. In order to explore the role of PrfA on Listeria biofilm development, we compared the abilities of biofilm formation by L. monocytogenes wild type strains (EGD and EGDe) and their prfA deletion mutants (EGD∆ prfA and EGDe∆ prfA ), nonpathogenic Listeria innocua , as well as the recombinant strains that express constitutively active mutant PrfA (PrfA*) in L. innocua (LI-pERL3- prfA *) and in EGDe∆ prfA (EGDe∆ prfA -pERL3- prfA *) at 37°C in brain heart infusion (BHI) medium using the polyvinyl chloride (PVC) microtiter plate assay and microscopic examination. Our results showed that the wild types of L. monocytogenes had strong abilities to develop biofilm with meshwork of bacterial aggregates, while biofilm with sparse small clumps were observed in L. innocua . The biofilm production of strains EGD∆ prfA and EGDe∆ prfA that lack funtional PrfA was reduced and could be recovered by the introduction of the PrfA*, however, the PrfA* had no impact on the biofilm forming ability of L. innocua . Our results suggest that PrfA plays a significant role in biofilm formation in L. monocytogenes but not in L. innocua , thus may reflect differences in the molecular mechanisms of biofilm formation by these two closely related species. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-011-9964-7 Authors Qingchun Zhou, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Central China Normal University, Luo Yu Road 152, Wuhan, 430079 People’s Republic of China Feifei Feng, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Central China Normal University, Luo Yu Road 152, Wuhan, 430079 People’s Republic of China Li Wang, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Central China Normal University, Luo Yu Road 152, Wuhan, 430079 People’s Republic of China Xiaoqin Feng, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Central China Normal University, Luo Yu Road 152, Wuhan, 430079 People’s Republic of China Xiaojiao Yin, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Central China Normal University, Luo Yu Road 152, Wuhan, 430079 People’s Republic of China Qin Luo, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Central China Normal University, Luo Yu Road 152, Wuhan, 430079 People’s Republic of China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Natural wild-type strains of Bacillus subtilis spore is regarded as a non-pathogenic for both human and animal, and has been classified as a novel food which is currently being used as probiotics added in the consumption. To identify B. subtilis spore proteins, we have accomplished a preliminary proteomic analysis of B. subtilis spore, with a combination of two-dimensional electrophoretic separations and matrix-assisted laser desorption ionization tandem time of flight mass spectrometry (MALDI–TOF–MS). In this article, we presented a reference map of 158 B. subtilis spore proteins with an isoelectric point (pI) between 4 and 7. Followed by mass spectrometry (MS) analysis, we identified 71 B. subtilis spore proteins with high level of confidence. Database searches, combined with hydropathy analysis and GO analysis revealed that most of the B. subtilis spore proteins were hydrophilic proteins related to catalytic function. These results should accelerate efforts to understand the resistance of spore to harsh conditions. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-011-9967-4 Authors Langyong Mao, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Shantong Jiang, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Bin Wang, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Liang Chen, School of Life Sciences, Sichuan University, 29# Wangjiang Road, Chengdu, 610064 Sichuan, People’s Republic of China Qin Yao, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Keping Chen, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    A fruity aroma-producing strain WG4 was isolated from a water sample collected from the Western Ghats, India. The 16S rRNA gene sequence analysis of strain WG4 indicated that Chryseobacterium indologenes , a member of the family ‘Flavobacteriaceae’ is the closest related species with a pair-wise sequence similarity of 98.6%. Strain WG4 produces a fruity aroma when grown on nutrient or trypticase soy agar plates. The fruity aroma is more when the strain WG4 is grown on agar plates compared to their growth in broth. The aromatic compounds produced by the strain WG4 were identified as ester compounds and were confirmed as ethyl-2-methylbutyrate and ethyl-3-methylbutyrate based on Gas Chromatography–Mass Spectrometry (GC–MS) analysis and using standard reference compounds. Even after repeated subcultures strain WG4 produced the same aroma in high intensity. Thus, strain WG4 could serve as a source for the production of these flavour compounds. Content Type Journal Article Pages 1-5 DOI 10.1007/s00284-011-9966-5 Authors P. Anil Kumar, Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, 500 007 India T. N. R. Srinivas, Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, 500 007 India A. R. Prasad, Indian Institute of Chemical Technology, Uppal Road, Hyderabad, 500 007 India S. Shivaji, Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, 500 007 India Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    This study reports the first detection of Wolbachia and yeast-like symbiont (YLS) harbored in Kerria lacca (Kerr), a scale insect, latter of which produces an economically important natural resin, known as lac. Wolbachia was detected using PCR amplification and sequencing of 16S rDNA; and further confirmation and phylogenetic analysis was carried out by fast evolving wsp gene. Neighbor-joining and maximum parsimonious (MP) analysis showed that this strain belongs to subgroup “ori” of Wolbachia super group B of arthropods. Wolbachia of K . lacca is hereby designated as “ w Kerlac” according to Wolbachia nomenclature system. Histological study revealed the presence of yeast-like endosymbiont, which was also confirmed by PCR amplification of 18S rDNA. Phylogenetic analysis revealed that YLS of K . lacca is quite distinct from YLS of aphid, planthoppers, and beetles. Putative roles of Wolbachia in lecanoid chromosome system of sex determination and in biased sex ratio of K . lacca populations; and YLS in nutritional supplementation and detoxifying substances which are deleterious to K . lacca , are hereby, suggested. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-011-9961-x Authors Amit Vashishtha, Department of Botany, University of Delhi, Delhi, 110007 India K. K. Sharama, Indian Institute of Natural Resin and Gum (IINRG), Namkum, Ranchi, India Suman Lakhanpaul, Department of Botany, University of Delhi, Delhi, 110007 India Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The genus Asaia (family Acetobacteraceae) was first introduced with a single species— Asaia bogorensis and later six more species were described namely A . siamensis , A . krungthepensis , A . lannaensis , A . platycodi , A . prunellae , and A . astilbes. Acetobacteraceae family has been divided into ten genera but, only three of them include nitrogen fixing species: Gluconacetobacter , Acetobacter , and Swaminathania . This article originated from our study primarily aimed to isolate new endosymbiotic nitrogen fixer among Acetobacteraceae during which we have isolated, for the first time in India, four different strains of Asaia sp. from three different sources: Michalia champaca flower, Anopheles mosquito, and ant Tetraponera rufonigra . All the endosymbiotic strains isolated possess the ability to fix nitrogen. Evidence for both nitrogenase activity and the presence of nifH gene in isolated Asaia sp. is presented. Asaia bogorensis (MTCC 4041 T ) and A . siamensis (MTCC 4042 T ), two of the validated type strains available from the repository, were tested positive for the presence of functional nitrogenase. The nifH gene sequences from these type strains were also confirmed and compared with other nitrogen fixing members of the family Acetobacteraceae. Our result corroborate with the previous reports that Asaia sp. are indeed widely distributed in nature but this is the first time demonstration of their functional nitrogenase activity. This study shows Asaia sp. as fourth genera of nitrogen fixing bacteria in the family Acetobacteraceae. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-011-9968-3 Authors Neeloy Samaddar, Department of Life Science & Biotechnology, Jadavpur University, Kolkata, India Arundhati Paul, Department of Life Science & Biotechnology, Jadavpur University, Kolkata, India Somnath Chakravorty, Department of Life Science & Biotechnology, Jadavpur University, Kolkata, India Writachit Chakraborty, Department of Life Science & Biotechnology, Jadavpur University, Kolkata, India Joydeep Mukherjee, School of Environmental Studies, Jadavpur University, Kolkata, India Debarati Chowdhuri, Department of Life Science & Biotechnology, Jadavpur University, Kolkata, India Ratan Gachhui, Department of Life Science & Biotechnology, Jadavpur University, Kolkata, India Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Leptothrix species in aquatic environments produce uniquely shaped hollow microtubules composed of aquatic inorganic and bacterium-derived organic hybrids. Our group termed this biologically derived iron oxide as “biogenous iron oxide (BIOX)”. The artificial synthesis of most industrial iron oxides requires massive energy and is costly while BIOX from natural environments is energy and cost effective. The BIOX microtubules could potentially be used as novel industrial functional resources for catalysts, adsorbents and pigments, among others if effective and efficient applications are developed. For these purposes, a reproducible system to regulate bacteria and their BIOX productivity must be established to supply a sufficient amount of BIOX upon industrial demand. However, the bacterial species and the mechanism of BIOX microtubule formation are currently poorly understood. In this study, a novel Leptothrix sp. strain designated OUMS1 was successfully isolated from ocherous deposits in groundwater by testing various culture media and conditions. Morphological and physiological characters and elemental composition were compared with those of the known strain L. cholodnii SP-6 and the differences between these two strains were shown. The successful isolation of OUMS1 led us to establish a basic system to accumulate biological knowledge of Leptothrix and to promote the understanding of the mechanism of microtubule formation. Additional geochemical studies of the OUMS1-related microstructures are expected provide an attractive approach to study the broad industrial application of bacteria-derived iron oxides. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-011-9957-6 Authors Michinori Sawayama, Department of Material Chemistry, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan Tomoko Suzuki, Department of Material Chemistry, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan Hideki Hashimoto, Department of Material Chemistry, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan Tomonari Kasai, Department of Material Chemistry, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan Mitsuaki Furutani, Department of Material Chemistry, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan Naoyuki Miyata, Department of Biological Environment, Akita Prefectural University, Akita, Japan Hitoshi Kunoh, Department of Material Chemistry, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan Jun Takada, Department of Material Chemistry, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    A Gram-negative, non-motile, catalase- and oxidase- positive, strictly aerobic, and short rod-shaped bacterium that was designated strain KOPRI 25157 T was isolated from coastal seawater sample in Antarctica. The temperature and pH ranges for growth on R2A agar were 10–20°C, and 5.0–10.0, respectively. Phylogenetic analyses of the 16S rRNA gene sequence of strain KOPRI 25157 T showed it to belong to the family Oxalobacteraceae of the class Betaproteobacteria , and it formed a distinct clade from other recognized members of the family. DNA G + C content was 65.9 mol%. Major ubiquinone was Q-8. Predominant cellular fatty acids were C 16:1 ω 7 c /15 iso 2OH (56.4%) and C 16:1 (30.5%). Major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, and unknown lipid. On the basis of these data, it is proposed that strain KOPRI 25157 T is the representative of a novel genus, for which the name Actimicrobium gen. nov. is proposed in the family Oxalobacteraceae. The type strain for Actimicrobium antarcticum sp. nov. is KOPRI 25157 T (=JCM 16673 T =KCTC 23040 T ). Content Type Journal Article Pages 1-5 DOI 10.1007/s00284-011-9962-9 Authors Eun Hye Kim, Division of Polar Life Sciences, Korea Polar Research Institute, Get-pearl Tower, 12 Gaetbeol-ro, Yeonsu-gu, Incheon, 406-840 Republic of Korea Hyun-Jeong Jeong, Division of Polar Life Sciences, Korea Polar Research Institute, Get-pearl Tower, 12 Gaetbeol-ro, Yeonsu-gu, Incheon, 406-840 Republic of Korea Yoo Kyoung Lee, Division of Polar Life Sciences, Korea Polar Research Institute, Get-pearl Tower, 12 Gaetbeol-ro, Yeonsu-gu, Incheon, 406-840 Republic of Korea Eun Young Moon, Institute of Microbiology, Seoul National University, Gwanak_1 Gwanak-ro, Gwanak-gu, Seoul, 151-742 Republic of Korea Jang-Cheon Cho, Division of Biology and Ocean Sciences, Inha University, 253 Yonghyun-dong, Nam-gu, Incheon, 402-751 Republic of Korea Hong Kum Lee, Division of Polar Life Sciences, Korea Polar Research Institute, Get-pearl Tower, 12 Gaetbeol-ro, Yeonsu-gu, Incheon, 406-840 Republic of Korea Soon Gyu Hong, Division of Polar Life Sciences, Korea Polar Research Institute, Get-pearl Tower, 12 Gaetbeol-ro, Yeonsu-gu, Incheon, 406-840 Republic of Korea Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Bromoxynil octanoate (BOO), the most widespread herbicide applied to maize, is potentially toxic to both animals and humans. In this article, a highly effective BOO-degrading bacterial strain, XB2, was isolated from the soil of a herbicide factory. The strain was identified as an Acinetobacter sp. based on its 16S rRNA gene sequence analysis, morphological, physiological, and biochemical properties. This strain could use BOO as its sole carbon source and could degrade 100 mg l −1 BOO to non-detectable levels in 72 h (h). The optimal pH and temperature for strain XB2’s growth and degradation of BOO in MSM are 7.0 and 30°C, respectively. We propose the following pathway of BOO degradation by strain XB2: the first step is the scission of the ester bond to form bromoxynil, bromoxynil then transformed to 3,5-dibromo-4-hydroxybenzoic acid due to the hydrolysis of nitriles, and debromination finally results in the formation of 3-bromo-4-hydroxybenzoic acid. Inoculating BOO-treated soil samples with strain XB2 resulted in a higher rate of BOO degradation than in non-inoculated soil, regardless of whether the soil had previously been sterilized. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-011-9965-6 Authors Tianming Cai, The College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, 210095 People’s Republic of China Liwei Chen, The College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, 210095 People’s Republic of China Jing Xu, The College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, 210095 People’s Republic of China Shu Cai, The College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, 210095 People’s Republic of China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The ability of Aspergillus fumigatus l -amino acid oxidase ( l -aao) to cause the resolution of racemic mixtures of dl -amino acids was investigated with dl -alanine, dl -phenylalanine, dl -tyrosine, and dl -aspartic acid. A chiral column, Crownpak CR+ was used for the analysis of the amino acids. The enzyme was able to cause the resolution of the three dl -amino acids resulting in the production of optically pure d -alanine (100% resolution), d -phenylalanine (80.2%), and d -tyrosine (84.1%), respectively. The optically pure d -amino acids have many uses and thus can be exploited industrially. This is the first report of the use of A. fumigatus l -amino acid oxidase for racemic resolution of dl -amino acids. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-011-9955-8 Authors Susmita Singh, Biotechnology Division, North East Institute of Science and Technology, Council of Scientific and Industrial Research, Jorhat, 785006 Assam, India Binod K. Gogoi, Biotechnology Division, North East Institute of Science and Technology, Council of Scientific and Industrial Research, Jorhat, 785006 Assam, India Rajib L. Bezbaruah, Biotechnology Division, North East Institute of Science and Technology, Council of Scientific and Industrial Research, Jorhat, 785006 Assam, India Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description: The endophytic actinomycete F4-20 was isolated from Tripterygium wilfordii Hook.f. and was confirmed to produce wilforgine, a secondary metabolite discovered in its host. F4-20 showed a close phylogenetic relationship to Streptomyces species. To seek elicitors that may enhance the production of wilforgine in F4-20, four plant stress molecules were applied to the in vitro liquid cultures. Results showed that methyl jasmonate (MeJA), salicylic acid (SA), and hydrogen peroxide (H 2 O 2 ) inhibited bacterial growth, whereas glutathione (GSH) treatment significantly increased bacterial growth. The wilforgine contents in the mycelia of F4-20 were reduced by MeJA and GSH but were induced by SA and H 2 O 2 . When added in the end of the culture period (7 day), 1 mM SA and 5 mM H 2 O 2 resulted in 69.35 ± 1.71 and 71.80 ± 3.35 µg/g DW of wilforgine production, 1.55 and 1.60 fold to that of control (44.83 ± 1.35 µg/g DW), respectively. Though this improved production was about 6.5 times lower than that of the natural root (454.00 µg/g dry root bark), it provided an alternative method for the production of valuable plant secondary metabolites.

Description: Magnetotactic bacteria synthesize intracellular magnetite and/or greigite magnetosome crystals. They play a significant role in both iron and sulfur cycles in sedimentary aquatic environments. To get insight into the bio-geochemical contribution of MTB, more studies concerning their ecology and their distribution in diverse habitats are necessary. The MTB community of an oil-industry polluted area of the French Mediterranean coast has been previously investigated. Here, we investigate the MTB community from coastal sediments of a Mediterranean pristine area using optical and transmission electron microscopy and phylogenetic analysis based on 16S rRNA gene sequences. A particularly high diversity of MTB was observed, with cocci phylogenetically distributed across the order Magnetococcales, including a novel cluster with sequences from the Mediterranean Sea designated as “Med group”, and novel morphotypes.

Description: A strictly aerobic, Gram-negative, beige-pigmented, short-rod-shaped, non-motile and chemoheterotrophic bacteria, designated K2-48 T was isolated from seawater collected in the Western North Pacific Ocean near Japan. Preliminary analysis based on the 16S rRNA gene sequence revealed that the novel isolate was affiliated with the family Oceanospirillaceae within the class Gammaproteobacteria and that it showed the highest sequence similarity (93.7 %) to Neptunomonas qingdaonensis P10-2-4 T . The strain could be differentiated phenotypically from recognized members of the family Oceanospirillaceae . The major fatty acids of strain K2-48 T were identified as summed feature 3 (C16:1 ω 7c and/or iso-C15:0 2-OH) and C16:0 as defined by the MIDI system. The DNA G+C content was determined to be 43.2 mol%, the major respiratory quinone was identified as ubiquinone 9 and a polar lipid profile was present consisting of phosphatidylethanolamine, a phosphatidylglycerol and an unidentified phospolipid. On the basis of polyphasic taxonomic studies, it was concluded that strain K2-48 T represents a novel genus sp. We propose the name Pelagitalea pacifica gen. nov., sp. nov. for this strain; its type strain is K2-48 T (=KCCM 90119 T ).

Description: Enterococcus faecalis has the ability to conjugate with the aid of aggregation substance (AS) and inducible sex pheromones to exchange genetic elements in food matrix. To evaluate the food safety condition and the transferable factor, 250 tetracycline-resistant food-borne E.   faecalis were collected in Korea. Among the isolates, a majority of tetracycline-resistant isolates (49.6 %) harbored both the tet (M) and tet (L) genes together, followed by tet (M) (19.6 %), and tet (L) (6.8 %) alone. Also, we found the combination of tet (L)/ tet (M)/ tet (O) or tet (M)/ tet (O). We identified two tet (S) genes including the isolate carrying tet (M) +  tet (S) genes. Additionally, most E. faecalis were positive for cpd and ccf (both 96.8 %) followed by cob (57.2 %). Through mating experiments, we confirmed E.   faecalis possessing the Int - Tn gene and/or any AS gene successfully transferred tet genes to JH2-2 E.   faecalis , whereas neither E.   faecalis carrying AS genes nor the Int - Tn gene showed the conjugation. Pulsed-field gel electrophoresis results supported a distinct pattern, implying transfer of genetic information. Our study revealed a high occurrence of tetracycline resistance genes in E.   faecalis from various foods. The widespread dissemination of tetracycline resistance genes would be promoted to transfer tetracycline resistance genes by pheromone-mediated conjugation systems.

Description: Successful colonization is the initial step for plant-bacteria interactions; therefore, the development of strategies to improve adherence to plant surfaces is critically important for environmental bacteria. Biofilm formation is thought to be one such strategy for bacteria to establish stable colonization on inert and living surfaces. Although biofilms play potential roles in enabling persistent bacterial colonization, little attention has been paid to biofilms formed by plant-associated bacteria. In this study, we characterized the biofilm-forming ability of 6 species of bacteria from the family Pseudomonadaceae: Pseudomonas protegens , Pseudomonas fluorescens , Pseudomonas putida , Pseudomonas stutzeri , Pseudomonas mendocina , and Pseudomonas syringae. These strains exhibit different degrees of biofilm formation depending on incubation time and nutrient availability. Distinct preferences for growth media were observed, as biofilms were formed by P. protegens with rich nutrients and by P. fluorescens and P. putida with poor nutrients. Likewise, P. stutzeri did not form biofilms with rich nutrients but did form biofilms under nutrient-poor conditions. These observations indicate that particular components in media may influence biofilm formation. P. putida , one of the strains with high biofilm-forming ability, showed the highest ability for initial attachment, which may be mediated by the hydrophobicity of its cell surface. P. mendocina also has high ability for initial attachment, and this strain produces cell surface-attached extracellular polysaccharides that promote cell aggregation. Thus, each strain possesses different properties that facilitate biofilm formation. Shedding light on bacterial strategies for colonization via biofilm formation would enable a better understanding of plant–bacteria interactions.

Description: The development of alternative energy sources by applying lignocellulose-based biofuel technology is critically important because of the depletion of fossil fuel resources, rising fossil fuel prices, security issues regarding the fossil fuel supply, and environmental issues. White-rot fungi have received much attention in recent years for their valuable enzyme systems that effectively degrade lignocellulosic biomasses. These fungi have powerful extracellular oxidative and hydrolytic enzymes that degrade lignin and cellulose biopolymers, respectively. Lignocellulosic biomasses from either agricultural or forestry wastes are abundant, low-cost feedstock alternatives in nature but require hydrolysis into simple sugars for biofuel production. This review provides a complete overview of the different lignocellulose biomasses and their chemical compositions. In addition, a complete list of the white-rot fungi-derived lignocellulolytic enzymes that have been identified and their molecular structures, mechanism of action in lignocellulose hydrolysis, and biochemical properties is summarized in detail. These enzymes include ligninolytic enzymes (laccase, manganese peroxidase, lignin peroxidase, and versatile peroxidase) and cellulolytic enzymes (endo-glucanase, cellobiohydrolase, and beta-glucosidase). The use of these fungi for low-cost lignocellulolytic enzyme production might be attractive for biofuel production.

Description: One of the issues that most concerns to both winemakers and producers of active dry yeasts is the stuck and sluggish fermentations of grape musts with high levels of sugar, reflecting the inability of inoculated yeast strain to complete the fermentation process. It is difficult to obtain a wine strain that possesses both adequate oenological and technological properties; thus, the correct approach to solving these problems is the application of breeding programs primarily focused on both properties. The first step toward this process is to characterize the phenotypic diversity between potential parental strains. In the present study, we have analyzed the fermentative behavior of 26 Saccharomyces cerevisiae wine strains in high-sugar conditions at 20 °C, using a range of tests, such as sporulation ability, spore viability, and tetrad analysis to determine the tolerance of these yeasts to several stress conditions. Most tested strains were homothallic and heterozygous for more than one character. Two auxotrophic derivatives with defects in amino acid or nucleic acid metabolism were obtained, and these strains could potentially be used for the development of hybridization techniques without using laboratory strains.

Description: Bacillus thuringiensis is a kind of insecticidal microorganism which can produce a variety of toxin proteins, it is particularly important to find an effective strategy to identify novel toxin proteins rapidly and comprehensively with the discovery of the wild-type strains. Multi-dimensional high-performance liquid chromatography combined with mass spectrometry has become one of the main methods to detect and identify toxin proteins and proteome of B. thuringiensis . In this study, protein samples from B. thuringiensis strain 4.0718 were analyzed on the basis of two-dimensional liquid chromatography–tandem mass spectrometry (2D-LC–MS/MS), and tryptic peptides of whole cell from the late sporulation phase were eluted at different concentration gradients of ammonium chloride and followed by secondary mass spectrum identification. 831 and 894 proteins were identified from two biological replicates, respectively, while 1,770 and 1,859 peptides were detected correspondingly. Among the identified proteins and peptides, 606 proteins and 1,259 peptides were detected in both replicates, which mean that 1,119 proteins and 2,370 peptides were unique to the proteome of this strain. A total of 15 toxins have been identified successfully, and seven of them were firstly discovered in B. thuringiensis strain 4.0718 that were Crystal protein (A1E259), pesticidal protein (U5KS09), Cry2Af1 (A4GVF0), Cry2Ad (Q9RM89), Cry1 (K4HMB5), Cry1Bc (Q45774), and Cry1Ga (Q45746). The proteomic strategy employed in the present study has provided quick and exhaustive identification of toxins produced by B. thuringiensis.

Description:    Bacterial FtsE gene codes for the ATP-binding protein, FtsE, which in complex with the transmembrane protein, FtsX, participates in diverse cellular processes. Therefore, regulated expression of FtsE and FtsX might be critical to the human pathogen, Mycobacterium tuberculosis , under stress conditions. Although ftsX gene of M. tuberculosis ( MtftsX ) is known to be transcribed from a promoter inside the upstream gene, ftsE , the transcriptional status of ftsE gene of M. tuberculosis ( MtftsE ) remains unknown. Therefore, the authors initiated transcriptional analyses of MtftsE , using total RNA from M. tuberculosis cells that were grown under stress conditions, which the pathogen is exposed to, in granuloma in tuberculosis patients. Primer extension experiments showed the presence of putative transcripts, T1, T2, T3, and T4. T1 originated from the intergenic region between the upstream gene, MRA_3135 , and MtftsE . T2 and T3 were found initiated from within MRA_3135 . T4 was transcribed from a region upstream of MRA_3135 . RT-PCR confirmed co-transcription of MRA_3135 and MtftsE . The cloned putative promoter regions for T1, T2, and T3 elicited transcriptional activity in Mycobacterium smegmatis transformants. T1, T2, and T3, but no new transcript, were present in the M. tuberculosis cells that were grown under the stress conditions, which the pathogen is exposed to in granuloma in tuberculosis patients. It showed lack of modulation of MtftsE transcripts under the stress conditions tested, indicating that ftsE may not have a stress response-specific function in M. tuberculosis . Content Type Journal Article Pages 1-9 DOI 10.1007/s00284-011-9897-1 Authors Sougata Roy, Indian Institute of Science, Microbiology and Cell Biology, Bangalore, Karnataka Srinivasan Vijay, Indian Institute of Science, Microbiology and Cell Biology, Bangalore, Karnataka Muthu Arumugam, Indian Institute of Science, Microbiology and Cell Biology, Bangalore, Karnataka Deepak Anand, Indian Institute of Science, Microbiology and Cell Biology, Bangalore, Karnataka Mushtaq Mir, Indian Institute of Science, Microbiology and Cell Biology, Bangalore, Karnataka Parthasarathi Ajitkumar, Indian Institute of Science, Microbiology and Cell Biology, Bangalore, Karnataka Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Cryptosporidium parvum , an intestinal apicomplexan parasite, is a significant cause of diarrheal diseases in both humans and animals. What is more, there is no promising strategy for controlling cryptosporidiosis. In this study, the P23 immunodominant surface protein of C. parvum sporozoites was stably expressed in the Lactobacillus casei Zhang strain and its immunogenicity was evaluated in a mouse model. The molecular weight (23 kDa) and immunogenicity of p23 gene expressed by L.   casei Zhang were similar to that of the native P23 protein. Oral immunization with control L.   casei Zhang and recombinant L. casei Zhang-p23 activated the mucosal immune system to elicit serum immunoglobulin G (IgG) and mucosal IgA in mice. Furthermore, the expression of cytokines such as IL-4, IL-6, and IFN-γ in splenocytes of mice was detected by real-time PCR after oral immunization. P23-specific immunocyte activation was also verified. These findings indicate that the live L.   casei Zhang vector may be a new tool for the production of mucosal vaccines against cryptosporidiosis in animals. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-011-9894-4 Authors Geriletu, Key Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, Inner Mongolia Agricultural University, 306 Zhaowuda Road, Hohhot, Inner Mongolia 010018, China Rihua Xu, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083 China Honglin Jia, National Research Center for Protozoan Disease, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokaido Japan Mohamad Alaa Terkawi, National Research Center for Protozoan Disease, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokaido Japan Xuenan Xuan, National Research Center for Protozoan Disease, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokaido Japan Heping Zhang, Key Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, Inner Mongolia Agricultural University, 306 Zhaowuda Road, Hohhot, Inner Mongolia 010018, China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Geldanamycin belongs to benzoquinone ansamycin antibiotic and has potent antitumor activities. In this study, a bacterial artificial chromosome (BAC) library with an average insert size of up to 150 kb was constructed from genomic DNA of Streptomyces autolyticus JX-47. A genetic-screening strategy was established using BAC end-sequencing and three pairs of primers designed to target the remote regions, gdmA1, gdmA3 and gdmRI, of the geldanamycin gene cluster. Three clones covering geldanamycin biosynthesis gene cluster were obtained, which together spanned a 250-kb genomic region, and a 150227-bp insert in the clone p4E9 was sequenced. Comparison with the reported geldanamycin gene cluster sequences from S. hygroscopicus revealed that it had the same gene arrangement and high gene homology in the polyketide synthase (PKS) region and its downstream with 84–100% DNA identity and 81–100% amino acid (AA) identity. Its DNA homology with the whole gene cluster sequence from S. hygroscopicus strain 17997 reached 99% identity. However, upstream of the PKS region exhibited great diversity, where only ORF16 was conserved, and the other genes including gdmL and gdmX were displaced. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-011-9940-2 Authors Shikun Dai, Key Laboratory of Marine Bio-Resources Sustainable Utilization, Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301 People’s Republic of China Yongchang Ouyang, Guangzhou Medical College, Guangzhou, 510182 People’s Republic of China Guanghua Wang, Key Laboratory of Marine Bio-Resources Sustainable Utilization, Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301 People’s Republic of China Xiang Li, Key Laboratory of Marine Bio-Resources Sustainable Utilization, Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301 People’s Republic of China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Phototrophic bacteria necessarily contain carotenoids for photosynthesis, and a few phototrophic purple bacteria accumulate unusual carotenoids. The carotenoids in the genera Phaeospirillum and Roseospira were identified using spectroscopic methods. All species of the genus Phaeospirillum contained characteristic polar carotenoids in addition to lycopene and hydroxylycopene (rhodopin); hydroxylycopene glucoside, dihydroxylycopene, and its mono- and/or diglucosides. From the structures of these carotenoids, their accumulation was suggested to be due to absence of CrtD (acyclic carotenoid C-3,4 desaturase) and to possession of glucosyltransferase. Species of the genus Roseospira have been reported to have unusual absorption spectra in acetone extract, and they were found to accumulate 3,4-didehydrorhodopin as a major carotenoid. This may be due to low activity of CrtF (acyclic 1-hydroxycarotenoid methyltransferase). The study concludes in identifying genus specific unusual carotenoids, which is probably due to characteristic nature of some carotenogenesis enzymes. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-011-9941-1 Authors Shinichi Takaichi, Department of Biology, Nippon Medical School, 297, Kosugi-cho 2, Nakahara, Kawasaki, 211-0063 Japan Takashi Maoka, Research Institute for Production Development, 15 Shimogamo Morimoto Cho, Sakyo ku, Kyoto, 606 0805 Japan Ch. Sasikala, Bacterial Discovery Laboratory, Centre for Environment, JNT University, Hyderabad, 500 085 India Ch. V. Ramana, Department of Plant Sciences, University of Hyderabad, Hyderabad, 500 046 India Keizo Shimada, Department of Biology, Tokyo Metropolitan University, Minami-ohsawa 1-1, Hachioji, Tokyo, 192-0397 Japan Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    A total of 136 samples of tap water were collected from state and municipal schools between March and November 2009. The samples were filtered through cellulose nitrate membranes that were seeded at non-nutrient agar 1.5% containing an overlayer of Escherichia coli suspension. Thirty-one (22.79%) tap water samples investigated were found positive for free-living amoebae (FLA). From these, 13 presented as FLA that seems to belong to the genus Acanthamoeba. All samples of FLA were cloned and identified as belonging to the genus Acanthamoeba by the morphology of cysts and trophozoites and by PCR using genus-specific primers that amplify the ASA.S1 region of 18S rDNA gene. Physiological tests of thermotolerance and osmotolerance were used to evaluate the pathogenicity of the isolates. The sequencing analysis by comparing the sequences submitted to GenBank, showed genotype distribution into groups T2, T2/T6, T6, and T4. In tests of thermotolerance and osmotolerance, 50% of the isolates had a low pathogenic potential. The results indicated the presence of Acanthamoeba in tap water in Rio Grande do Sul, Brazil, revealing its importance and the need for more epidemiological studies to determine their distribution in the environment and its pathogenic potential. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-011-0003-5 Authors Mari Aline Todero Winck, Departamento de Microbiologia, Imunologia e Parasitogia, Instituto de Ciências Básicas da Saúde, Setor de Parasitologia, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite, 500, Porto Alegre, RS 90050-170, Brazil Karin Caumo, Departamento de Microbiologia, Imunologia e Parasitogia, Instituto de Ciências Básicas da Saúde, Setor de Parasitologia, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite, 500, Porto Alegre, RS 90050-170, Brazil Marilise Brittes Rott, Departamento de Microbiologia, Imunologia e Parasitogia, Instituto de Ciências Básicas da Saúde, Setor de Parasitologia, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite, 500, Porto Alegre, RS 90050-170, Brazil Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The potential use of Brettanomyces anomalus PSY-001 as an additional starter culture for the production of Rice-steamed sponge cake (RSSC), a traditional fermented food in China, was investigated. Two productions of RSSC, each containing batches of experimental cakes with Brettanomyces added and reference cakes with the leavened liquid added were carried out. For both experimental and reference cakes, chemical analysis and sensory evaluation were carried out during the fermentation period. The results showed that experimental cakes had desirable aroma and taste. The observed differences indicate a positive contribution to the overall quality of RSSC by B. anomalus PSY-001. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-011-9997-y Authors Peng Wu, College of Food Science and Technology, Huazhong Agricultural University, 430070 Wuhan, China Xiaoyun Xu, College of Food Science and Technology, Huazhong Agricultural University, 430070 Wuhan, China Yongxia Xu, College of Food Science and Technology, Huazhong Agricultural University, 430070 Wuhan, China Qingchan Chen, College of Food Science and Technology, Huazhong Agricultural University, 430070 Wuhan, China Siyi Pan, College of Food Science and Technology, Huazhong Agricultural University, 430070 Wuhan, China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    To study the prevalence and isoforms of the pathogenicity island ETT2 among pathogenic Escherichia coli , as well as to determine the relationship between the ETT2 locus and other virulence factors, PCR amplifications target to the 35 ETT2-associated genes were established and used to investigate the presence of the ETT2 locus in 168 E. coli isolates from weaned piglets with edema and/or diarrhea or dairy cows with mastitis. The results showed that the ETT2 locus could be identified in the pathogenic E. coli isolates from colibacillosis in pigs and in the ones from mastitis in cows, but the presence of ETT2 among the isolates of porcine origin were significantly higher (85.87%) than that (47.37%) of bovine origin. Furthermore, 11 ETT2 isoforms were found in this research, including an intact form and 10 deletion types. The intact ETT2 was the prevalent form among the pathogenic E. coli isolates of porcine origin, and highly associated with the presence of shigatoxin type 2e (Stx2e), while the great majority isolates of bovine origin just carried various deletion types, and no distinct association with other virulence factors, e.g., the presence/absence of LT1, ST2, Cnf2, Tra, HPI, Hly, and F17a fimbriae. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-011-0032-0 Authors DaRong Cheng, College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009 China ShanYuan Zhu, Jiangsu Animal Husbandry and Veterinary College, Taizhou, 225300 China ZhiRui Su, College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009 China WeiYong Zuo, Jiangsu Animal Husbandry and Veterinary College, Taizhou, 225300 China Hui Lu, Jiangsu Animal Husbandry and Veterinary College, Taizhou, 225300 China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    A collection of 94 unusual members of the Enterobacteriaceae were screened for the presence of extended spectrum β-lactamases (ESBLs) using the MicroScan ESβL plus dried confirmation panel. Presumptively positive strains were then confirmed for the presence of an ESBL by double disk diffusion, E-test strips (AB Biodisk, Solna, Sweden) and PCR for SHV, TEM, and CTX-M2 genes. Of the 18 strains initially positive on the ESβL panel only three strains ( Leminorella grimontii , Klebsiella ozaenae , and Kluyvera ascorbata ) were positive by confirmation methods. These results suggest laboratories should be cautious regarding the methodology employed in screening for the presence of ESBLs in enteric bacteria. However, it should be noted that of the 94 strains, 29 were found to be resistant to two or more of the antibiotics present in the MicroScan ESβL plus panel indicating that there are potential treatment issues with these organisms despite their lack of ESBLs. Content Type Journal Article Pages 1-4 DOI 10.1007/s00284-011-0057-4 Authors Sharon L. Abbott, Microbial Diseases Laboratory, CA State Department of Public Health, Richmond, CA, USA Janice A. Lidgard, Microbial Diseases Laboratory, CA State Department of Public Health, Richmond, CA, USA Wendy K. W. Cheung, Microbial Diseases Laboratory, CA State Department of Public Health, Richmond, CA, USA Martha N. Obeso, Microbial Diseases Laboratory, CA State Department of Public Health, Richmond, CA, USA Zenda L. Berrada, Microbial Diseases Laboratory, CA State Department of Public Health, Richmond, CA, USA J. Michael Janda, Microbial Diseases Laboratory, CA State Department of Public Health, Richmond, CA, USA Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Pantoea agglomerans YS19 is a rice endophytic bacterium characterized to form multicellular biofilm-like structures called symplasmata. Phenotypic distinctions between symplasmata-forming cells and planktonic cells are crucial for understanding YS19’s survival strategies. In this study, a 43.1 kDa protein SPM43.1 was identified to show significant resistance to the aggregation effect caused by denaturing acidic conditions. MALDI-TOF analysis data indicated that it is a maltose-binding protein homolog while contains sequence homologous to the chaperone protein, ClpB. The purified SPM43.1 protein was detected to exhibit chaperone-like activities at acidic conditions, where its conformation transformed from an ordered to a globally less ordered structure as revealed by circular dichroism spectroscopy, showing a similar property to most chaperone proteins. The expression of SPM43.1 in YS19 is initiated when bacterial cells begin to aggregate, yet its amount in planktonic cells greatly exceeds that in symplasmata-forming cells, suggesting its crucial role to the survival of planktonic cells in experiencing environmental fluctuations. However, the bacterium prefers to form symplasmata, while not to express SPM43.1 proteins, for surviving the artificially set fluctuant (acid here) environments. This study provides valuable information on the life styles and survival strategies of microorganisms that forms multicellular aggregates at specific growth stages. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-011-0055-6 Authors Qianqian Li, School of Life Science, Beijing Institute of Technology, #5 Zhongguancun Nandajie, Beijing, 100081 People’s Republic of China Yuxuan Miao, School of Life Science, Beijing Institute of Technology, #5 Zhongguancun Nandajie, Beijing, 100081 People’s Republic of China Ting Yi, School of Life Science, Beijing Institute of Technology, #5 Zhongguancun Nandajie, Beijing, 100081 People’s Republic of China Jia Zhou, School of Life Science, Beijing Institute of Technology, #5 Zhongguancun Nandajie, Beijing, 100081 People’s Republic of China Zhenyue Lu, School of Life Science, Beijing Institute of Technology, #5 Zhongguancun Nandajie, Beijing, 100081 People’s Republic of China Yongjun Feng, School of Life Science, Beijing Institute of Technology, #5 Zhongguancun Nandajie, Beijing, 100081 People’s Republic of China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Sediments from Xuanwu Lake have been dredged in the past 3 years to improve the water quality, but methanogenesis should still exist in the newly settled sediment. Methane production, methanogens, and physiochemical parameters were detected in the surface sediments (0–5 cm) and/or vertical sediments (0–21 cm, segmented at interval of 3 cm). Methane flux at water–air interface varied among five detected sites. Principal component analysis showed that CH 4 flux, content of water and the concentration of total nitrogen (TN), CH 4 and organic matters (OM) weighed most heavily on the component I in surface sediments while different patterns were observed for vertical sediments. The copy number of the 16S rRNA gene for bacteria was lower in the surface sediment (0–6 cm) than that in deeper sediments (12–21 cm), while 16S rRNA genes of Archaea were almost evenly distributed in the vertical sediments. Representatives belonging to the orders Methanobacteriales , Methanomicrobiales , and Methanosarcinales were detected in all samples of the vertical sediments, except that no members of the Methanococcales were detected in the samples at 0–6 cm. The level of Methanobacteriales reached a highest density at 18.1 × 10 4  copies g −1  dry weight (dw) at 6–9 cm; for Methanosarcinales (76.89 × 10 6  copies g −1  dw) and Methanococcales (82.70 × 10 3  copies g −1  dw) at 12–15 cm, whereas for Methanomicrobiales (43.37 × 10 6  copies g −1  dw) at 9–12 cm. Methanosarcinaceae and Methanosaetaceae reached to their highest densities at 6–9 and 9–12 cm, respectively. These data provided useful information for better understanding the methanogenesis in the newly settled sediments of a recently dredged lake. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0103-x Authors Dong-Lin Zhu, The State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University, 163 Xianlin Avenue, Nanjing, 210046 Jiangsu, China Cheng Sun, The State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University, 163 Xianlin Avenue, Nanjing, 210046 Jiangsu, China Huan He, The State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University, 163 Xianlin Avenue, Nanjing, 210046 Jiangsu, China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Hexazinone, a triazine herbicide that is often detected as a ground and surface water contaminant, inhibits electron transport in photosynthetic organisms and is toxic to primary producers that serve as the base of the food chain. This laboratory study evaluated the ability of two types of microbial reactors, i.e., a vegetable oil-based nitrogen-limiting biobarrier and an aerobic slow sand filter, as methods for removing hexazinone from simulated groundwater. The N-limiting biobarriers degraded hexazinone, but did so with a 52 week incubation period and a removal efficiency that varied greatly among replicates, with one biobarrier showing a removal efficiency of ~95% and the other an efficiency of ~50%. More consistent degradation was obtained with the aerobic sand biobarriers. Four aerobic biobarriers were evaluated and all behaved in a similar manner degrading hexazinone with removal efficiencies of ~97%; challenging two of the aerobic biobarriers with large amounts of influent hexazinone showed that these barriers are capable of efficiently remediating large amounts (〉100 mg L −1 ) of hexazinone at high efficiency. The remediation process was due to biological degradation rather than abiotic processes. The long lag phase observed in both types of reactors suggests that an acclimation process, where microorganisms capable of degrading hexazinone increased in numbers, was required. Also, the isolation of bacteria that show a positive growth response to the presence of hexazinone in their growth media suggests biological degradation. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0086-7 Authors William J. Hunter, USDA–ARS, 2150-D Centre Avenue, Fort Collins, CO 80526-8119, USA Dale L. Shaner, USDA–ARS, 2150-D Centre Avenue, Fort Collins, CO 80526-8119, USA Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Little is known about the association among the transcription, post-transcription, and protein production of the fumA gene. This study demonstrates that increasing growth rate ( k ) from 0.24/h to 0.96/h causes a marked eightfold reduction in fumA transcription as assessed using the β-galactosidase activity from fumA promoter fused with a lacZ reporter. It was further confirmed using Northern blot analysis. Most interestingly, the FumA protein levels remained unchanged over the growth rate, as indicated by Western blot analysis. Therefore, whether the reduced fumA mRNA expression under the high growth rate can be overcome by increasing the stability of the fumA mRNA was tested. The half-life of fumA mRNA was established to significantly increase by fivefold when the growth rate was increased to 0.96/h. This finding suggests that the cells could turn down the expression of fumA mRNA because of increased stability of its mRNA under the high growth rate. This notion indicates that mRNA stability plays an essential role in maintaining a critical cellular level of a given protein when the mRNA transcript is downregulated by a metabolic event. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0087-6 Authors Hsiao-Hsien Lin, Department of Biological Science and Technology, College of Biological Science and Technology, National Chiao Tung University, 75 Po-Ai Street, Hsinchu, 30050 Taiwan, ROC Ching-Hsueh Lin, Department of Biological Science and Technology, College of Biological Science and Technology, National Chiao Tung University, 75 Po-Ai Street, Hsinchu, 30050 Taiwan, ROC Shiaw-Min Hwang, Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu, Taiwan, ROC Ching-Ping Tseng, Department of Biological Science and Technology, College of Biological Science and Technology, National Chiao Tung University, 75 Po-Ai Street, Hsinchu, 30050 Taiwan, ROC Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Mycoplasma mobile, a pathogen of freshwater fish, glides easily across surfaces, colonizes on the fish gill, and causes necrosis. The cell surface is differentiated into three parts: the head, neck, and body. Mobile variable surface proteins (Mvsps) localizing at each of these parts may be involved in surface variation including phase variation and antigenic variation, although no proof exists. In this study, we examined this possibility by focusing on MvspI, the largest Mvsp. Immunofluorescence microscopy showed that MvspI is expressed on the surfaces of all cells. When anti-MvspI antibody was added at concentrations over 0.8 nM, MvspI was observed to decrease over time. After 72 h of cultivation with the antibody, the fluorescence intensity and amount of MvspI decreased up to 13 and 39%, respectively, compared to those of cells grown without antibody. These changes were reversed by the removal of the antibody. Such effects were not observed when another antibody targeting other Mvsps was used, suggesting that the decrease is specific to the relationship between MvspI and the antibody. Cell growth was also inhibited by the antibody, but the decrease in MvspI could not be explained by the selective growth of MvspI-negative variants or by the inhibition of growth with other conditions. The decrease in MvspI caused by the antibody binding may suggest a novel type of surface variation, designated here as “mycoplasmal antigen modulation.” Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-012-0090-y Authors Heng Ning Wu, Department of Biology, Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka, 558-8585 Japan Chie Kawaguchi, Department of Biology, Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka, 558-8585 Japan Daisuke Nakane, Department of Biology, Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka, 558-8585 Japan Makoto Miyata, Department of Biology, Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka, 558-8585 Japan Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    A Gram-positive aerobic rod-shaped non-motile bacterium designated A23 T was isolated from bamboo extract that had been used to remove odor and was characterized to determine its taxonomic position. 16S rRNA gene sequence analysis revealed that strain A23 T belongs to the phylum Actinobacteria. The highest degree of sequence similarities was determined to be with Leucobacter salsicius M1-8 T (96.7%), Leucobacter exalbidus K-540B T (96.4%), Leucobacter chromiireducens subsp. chromiireducens L-1 T (96.4%), Leucobacter komagatae IFO 15245 T (96.4%) and Leucobacter aerolatus Sj10 T (96.4%). Chemotaxonomic data revealed that strain A23 T possesses menaquinone MK11, and its cell wall peptidoglycan contained 2,4-diaminobutyric acid, alanine, glycine, glutamic acid and γ-aminobutyric acid. The polar lipid profile of strain A23 T contained diphosphatidylglycerol, phosphatidylglycerol and an unknown glycolipid. The predominant fatty acids were iso-C 16:0 (31.5%), anteiso-C 15:0 (43.2%) and anteiso-C 17:0 (13.9%), all of which corroborated the assignment of the strain to the genus Leucobacter. Based on these data, A23 T (=KEMC 551-022 T  = JCM 17538 T ) should be classified as the type strain for a novel Leucobacter species, for which the name Leucobacter margaritiformis sp. nov. is proposed. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-012-0089-4 Authors Jin-Ha Lee, Division of Natural Science, Department of Bioengineering, Kyonggi University, 94-6 Iui-dong, Yeongtong-gu, Suwon, 433-760 Republic of Korea Sang-Seob Lee, Division of Natural Science, Department of Life Science, Kyonggi University, 94-6 Iui-dong, Yeongtong-gu, Suwon, 433-760 Republic of Korea Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    We report the characterization of a small cryptic plasmid unlike any previously described from Moraxella bovis ATCC 10900, a Gram-negative bacterium belonging to the family Moraxellaceae. The complete nucleotide sequence of the plasmid pMbo4.6 was determined. The plasmid was analyzed and found to be 4658 in size with a G+C content of 38.6 mol %. Computer analysis of the sequence data revealed four major open reading frames encoding putative proteins of 10.1 (ORF1), 64.2 (ORF2), 45.7 (ORF3), and 12.1 kDa (ORF4). ORF1 and ORF2 encode proteins that show a high level of amino acid sequence similarity (44 %) with some mobilization proteins. ORF3 encodes a protein showing a relatively high amino acid sequence similarity (about 40 %) with several plasmid replication initiator proteins. Upstream of ORF3, a 320-bp intergenic region, constituting the putative origin of replication that contained an AT-rich region followed by four direct repeats, was identified. This set of repeated sequences resembles iteron structures and plays an important role in the control of plasmid replication by providing a target site for the initiation of transcription and replication factors (IHF and RepA). Several palindromic sequences, inverted repeats, and hairpin-loop structures, which might confer regulatory effects on the replication of the plasmid, were also noted. ORF4 encodes an uncharacterized protein, conserved in bacteria, belonging to the DUF497 family. Sequence analysis and structural features indicate that pMbo4.6 replicates by a theta mechanism. Content Type Journal Article Pages 1-9 DOI 10.1007/s00284-012-0257-6 Authors Beata Furmanek-Blaszk, Department of Microbiology, University of Gdansk, Kladki 24, 80-822 Gdansk, Poland Natalia Kurpiewska, Department of Microbiology, University of Gdansk, Kladki 24, 80-822 Gdansk, Poland Robert Boratynski, Department of Microbiology, University of Gdansk, Kladki 24, 80-822 Gdansk, Poland Marian Sektas, Department of Microbiology, University of Gdansk, Kladki 24, 80-822 Gdansk, Poland Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Klebsiella bacteria have emerged as an increasingly important cause of community-acquired nosocomial infections. Extensive use of broad-spectrum antibiotics in hospitalised patients has led to both increased carriage of Klebsiella and the development of multidrug-resistant strains that frequently produce extended-spectrum β-lactamases and/or other defences against antibiotics. Many of these strains are highly virulent and exhibit a strong propensity to spread. In this study, six lytic Klebsiella bacteriophages were isolated from sewage-contaminated river water in Georgia and characterised as phage therapy candidates. Two of the phages were investigated in greater detail. Biological properties, including phage morphology, nucleic acid composition, host range, growth phenotype, and thermal and pH stability were studied for all six phages. Limited sample sequencing was performed to define the phylogeny of the K. pneumoniae - and K. oxytoca -specific bacteriophages vB_Klp_5 and vB_Klox_2, respectively. Both of the latter phages had large burst sizes, efficient rates of adsorption and were stable under different adverse conditions. Phages reported in this study are double-stranded DNA bacterial viruses belonging to the families Podoviridae and Siphoviridae . One or more of the six phages was capable of efficiently lysing ~63 % of Klebsiella strains comprising a collection of 123 clinical isolates from Georgia and the United Kingdom. These phages exhibit a number of properties indicative of potential utility in phage therapy cocktails. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-012-0264-7 Authors Natia Karumidze, Eliava Institute of Bacteriophages, Microbiology and Virology, Tbilisi, Georgia Ia Kusradze, Eliava Institute of Bacteriophages, Microbiology and Virology, Tbilisi, Georgia Sophio Rigvava, Eliava Institute of Bacteriophages, Microbiology and Virology, Tbilisi, Georgia Marine Goderdzishvili, Eliava Institute of Bacteriophages, Microbiology and Virology, Tbilisi, Georgia Kumar Rajakumar, Department of Infection, Immunity and Inflammation, University of Leicester, Leicester, LE1 9HN UK Zemphira Alavidze, Eliava Institute of Bacteriophages, Microbiology and Virology, Tbilisi, Georgia Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Currently the treatment of Mycobacterium tuberculosis (TB) infection is largely limited due to the prevalence of multidrug resistance strains. Over-expressing the efflux pumps such as the ATP-binding cassette (ABC) transporter has been reported to significantly contribute to its resistance to several antibiotics. This study investigated the expression profile of one important ABC efflux pump, Rv1217c–Rv1218c, by quantitative real-time PCR (RT-qPCR) in clinical isolates from China, which also revealed its association with the multidrug resistance of M. tuberculosis . Significantly increased expressions of Rv1217c and Rv1218c at transcriptional level have been observed in multidrug-resistant TB group (MDR-TB) compared to those of the drug-susceptible group ( P  〈 0.05), when H37Rv strain was used as the control. Furthermore, correlation analysis revealed that the over-expression of both Rv1217c and Rv1218c resulted in the higher minimum inhibition concentrations (MICs) of rifampicin (RIF) (OR = 1.01, P  〈 0.05 of Rv1217c; OR = 1.23, P  〈 0.05 of Rv1218c), while the over-expression of Rv1218c only led to the higher MICs of isoniazid (INH) (OR = 1.17, P  〈 0.05). Our findings contributed to the better understanding of the molecular mechanisms of ABC efflux pumps, in particular Rv1217c–Rv1218c, in M. tuberculosis and will assist in developing new antibiotic treatments for multidrug-resistant M. tuberculosis in the future. Content Type Journal Article Pages 1-5 DOI 10.1007/s00284-012-0215-3 Authors Ke Wang, Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, 214063 Jiangsu Province, China Hao Pei, Wuxi Hospital of Infectious Disease, Wuxi, 214005 Jiangsu Province, China Biao Huang, Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, 214063 Jiangsu Province, China Xue Zhu, Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, 214063 Jiangsu Province, China Jue Zhang, Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, 214063 Jiangsu Province, China Bin Zhou, Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, 214063 Jiangsu Province, China Lan Zhu, Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, 214063 Jiangsu Province, China Yi Zhang, Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, 214063 Jiangsu Province, China Fan-Fan Zhou, Faculty of Pharmacy, University of Sydney, NSW, 2006 Australia Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The potential for the transport and diffusion of some pathogenic microorganisms by migratory birds is of concern. Migratory birds may be involved in the dispersal of microorganisms and may play a role of mechanical and biological vectors. The efficiency of dispersal of pathogenic microorganisms depends on a wide range of biotic and abiotic factors that influence the survival or disappearance of a given agent in a geographical area. In the present study, 349 migratory birds were captured in four sites (Mazara del Vallo, Lampedusa, Ustica and Linosa), representing the main stop-over points during spring and autumnal migration, and analyzed for the presence of filamentous fungi. A total of 2,337 filamentous fungi were isolated from 216 birds and identified by a combined phenotypic-genotypic approach to species level. Twelve species were identified in the study, with Cladosporium cladosporioides , Alternaria alternata , and Aspergillus niger as the most abundant. The transport of these fungal species isolated in this study is of considerable importance because some of these species can create dangers to human health. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0262-9 Authors Antonio Alfonzo, DEMETRA Department, Università degli Studi di Palermo, Viale delle Scienze 4, 90128 Palermo, Italy Nicola Francesca, DEMETRA Department, Università degli Studi di Palermo, Viale delle Scienze 4, 90128 Palermo, Italy Ciro Sannino, DEMETRA Department, Università degli Studi di Palermo, Viale delle Scienze 4, 90128 Palermo, Italy Luca Settanni, DEMETRA Department, Università degli Studi di Palermo, Viale delle Scienze 4, 90128 Palermo, Italy Giancarlo Moschetti, DEMETRA Department, Università degli Studi di Palermo, Viale delle Scienze 4, 90128 Palermo, Italy Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    In this study, more than 150 bacteria showing antagonistic properties against bacterial and fungal pathogens of the tomato plant were isolated and characterized. The most efficient agents against these phytopathogenic microorganisms belong to the genus Bacillus : the best biocontrol isolates were representatives of Bacillus subtilis , B. mojavensis and B. amyloliquefaciens species. They intensively produced fengycin or/and surfactin depsipeptide antibiotics and also proved to be excellent protease secretors. It was proved, that the selected strains were able to use ethylenethiourea (ETU) as sole nitrogen source. These antagonistic and ETU-degrading Bacillus strains can be applied as biocontrol and also as bioremediation agents. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-012-0263-8 Authors Csaba Vágvölgyi, Department of Microbiology, Faculty of Science and Informatics, University of Szeged, Közép fasor 52, Szeged, 6726 Hungary Enikő Sajben-Nagy, Department of Microbiology, Faculty of Science and Informatics, University of Szeged, Közép fasor 52, Szeged, 6726 Hungary Bettina Bóka, Department of Microbiology, Faculty of Science and Informatics, University of Szeged, Közép fasor 52, Szeged, 6726 Hungary Mónika Vörös, Department of Microbiology, Faculty of Science and Informatics, University of Szeged, Közép fasor 52, Szeged, 6726 Hungary Adrienn Berki, Department of Microbiology, Faculty of Science and Informatics, University of Szeged, Közép fasor 52, Szeged, 6726 Hungary Andrea Palágyi, Department of Microbiology, Faculty of Science and Informatics, University of Szeged, Közép fasor 52, Szeged, 6726 Hungary Judit Krisch, Faculty of Engineering, Institute of Food Engineering, University of Szeged, Mars tér 7, Szeged, 6724 Hungary Biljana Škrbić, Faculty of Technology, University of Novi Sad, Bulevar cara Lazara 1, 21000 Novi Sad, Serbia N. Đurišić-Mladenović, Faculty of Technology, University of Novi Sad, Bulevar cara Lazara 1, 21000 Novi Sad, Serbia László Manczinger, Department of Microbiology, Faculty of Science and Informatics, University of Szeged, Közép fasor 52, Szeged, 6726 Hungary Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Heavy metal resistance microorganism plays an important role on polluted soil bioremediation. To obtain further knowledge of the resistant mechanism employed by cadmium-resistant bacteria, some gene expression profiles at transcription level were investigated in P. aeruginosa E 1 subjected to cadmium stress using real-time PCR. Exposure to cadmium for 1 h, the expression of czc A, czc B, and czc C all reached the peak of up-regulation 8.82-, 4.83-, and 7.43-fold, respectively. The response of czc D was earlier and stronger than czc ABC. Cys M contributed to cysteine synthesis kept up-regulation within the beginning 2 h. The expression of mgt AE genes related to Mg 2+ influx was up-regulated all the while, znuB responsible for Zn 2+ transportation kept up-regulation from 30 min to 4 h. The result support the two cadmium-resistance mechanisms including effluxing and inactive the heavy metal ions. The mechanism was brought that increase of Mg 2+ and Zn 2+ in cytoplasm would prevent Cd 2+ -binding enzymes to decrease the harm to cell. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0224-2 Authors Xiaoxi Zeng, Hunan Key Lab of Green Packaging and Biological Nanotechnology, Hunan University of Technology, Zhuzhou, 412007 People’s Republic of China Jianxin Tang, Hunan Key Lab of Green Packaging and Biological Nanotechnology, Hunan University of Technology, Zhuzhou, 412007 People’s Republic of China Xueduan Liu, School of Minerals Processing and Bioengineering, Central South University, Changsha, 410083 People’s Republic of China Pei Jiang, Hunan Key Lab of Green Packaging and Biological Nanotechnology, Hunan University of Technology, Zhuzhou, 412007 People’s Republic of China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Listeria monocytogenes is an intracellular bacterium responsible for listeriosis in both humans and animals. Infected livestock is believed to be one source of this pathogen. Vaccination is an optimal approach to control the occurrence of this disease in livestock. However, inactivated vaccines have been reported to be insufficient to offer immune protection against L. monocytogenes . Here we evaluated the immune protection capacity of a combination of recombinant p60 and LLO. Mice immunized with p60 and LLO generated a high level of anti- L. monocytogenes antibodies. In addition, the elevated levels of IFN-γ and the decreased levels of IL-4 were also observed in these treated mice. Consistent with the colonization of L. monocytogenes post infection, all mice in the control group died within 5 days after infection of L. monocytogenes , while 40, 40, 80, and 100 % of animals immunized with inactivated L. monocytogenes vaccine (ILMV), LLO + ILMV, p60 + ILMV, and p60 + LLO + ILMV, respectively, survived for 2 weeks. Collectively, the results presented in this study demonstrate the capacity of a combination of LLO and p60 to elicit high protective immune responses against L. monocytogenes infection. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0238-9 Authors Xuenong Luo, Key Laboratory of Zoonoses of CAAS, Key Laboratory of Veterinary Parasitology of Gansu Province, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, CAAS, Xujiaping 1, Yanchangbu, Lanzhou, 730046 Gansu, China Xuepeng Cai, Key Laboratory of Zoonoses of CAAS, Key Laboratory of Veterinary Parasitology of Gansu Province, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, CAAS, Xujiaping 1, Yanchangbu, Lanzhou, 730046 Gansu, China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Glycerol and glucose fermentation redox routes by Escherichia coli and their regulation by oxidizing and reducing reagents were investigated at different pHs. Cell growth was followed by decrease of pH and redox potential ( E h ). During glycerol utilization at pH 7.5 ∆pH, the difference between initial and end pH, was lower compared with glucose fermentation. After 8 h growth, during glycerol utilization E h dropped down to negative values (−150 mV) but during glucose fermentation it was positive (+50 mV). In case of glycerol H 2 was evolved at the middle log phase while during glucose fermentation H 2 was produced during early log phase. Furthermore, upon glycerol utilization, oxidizer potassium ferricyanide (1 mM) inhibited both cell growth and H 2 formation. Reducing reagents dl -dithiothreitol (3 mM) and dithionite (1 mM) inhibited growth but stimulated H 2 production. The findings point out the importance of reductive conditions for glycerol fermentation and H 2 production by E. coli . Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0240-2 Authors Anna Poladyan, Department of Biophysics, Biology Faculty, Yerevan State University, 1 A. Manoukian Street, 0025 Yerevan, Armenia Arev Avagyan, Department of Microbiology, Plants and Microbes Biotechnology, Biology Faculty, Yerevan State University, 1 A. Manoukian Street, 0025 Yerevan, Armenia Anait Vassilian, Department of Ecology and Nature Protection, Biology Faculty, Yerevan State University, 1 A. Manoukian Street, 0025 Yerevan, Armenia Armen Trchounian, Department of Biophysics, Biology Faculty, Yerevan State University, 1 A. Manoukian Street, 0025 Yerevan, Armenia Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Photorhabdus are motile Gram-negative bacteria that have a mutualistic association with Heterorhabditis nematodes (Heterorhabditidae). These bacteria possess peculiar biochemical characteristics such as inability to reduce nitrates, and the capacity to ferment only a limited number of carbohydrates. Heterorhabditis nematodes vector the bacteria from one insect host to another and also provide shelter to the bacteria from soil stressors and antagonists. Once inside the insect host, the bacterial symbionts are released and produce toxins and secondary metabolites and broad-spectrum antibiotics, which kill the host by septicemia within 48 h. At present, three Photorhabdus spp. have been identified: P. luminescens , P. temperata , and P. asymbiotica , and many subspecies have also been described. Characterization of new species and subspecies has been based on sequence data, mostly of the 16S rDNA, and also of a selection of protein coding genes. In addition to this, phenotypic traits including temperature growth, colony morphology, color, light production, carbohydrate response, and assimilation, among others, have been considered. In this study, we characterize the bacterial symbiont of Heterorbabditis sonorensis , a recently discovered entomopathogenic nematode species form the Sonoran desert in Arizona, USA. A selection of classic biochemical and molecular methods including sequence data of six genes: 16s rDNA, and four protein coding genes: gyr B, recA, glt X, and dna N were considered. Evolutionary relationships of this new Photorhabdus subsp. were inferred considering maximum parsimony and Bayesian analyses. Content Type Journal Article Pages 1-10 DOI 10.1007/s00284-012-0220-6 Authors Rousel A. Orozco, Department of Entomology, The University of Arizona, 1140 E. South Campus Dr., Tucson, AZ 85721-0036, USA Tara Hill, Department of Chemistry and Biochemistry, The University of Arizona, BSW 360, P.O. Box 210088, Tucson, AZ, USA S. Patricia Stock, Department of Entomology, The University of Arizona, 1140 E. South Campus Dr., Tucson, AZ 85721-0036, USA Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The mechanisms for the enhancement of pristinamycin production in the high-yielding recombinants of Streptomyces pristinaespiralis obtained by genome shuffling were investigated by quantitative real-time PCR (Q-PCR) and amplified fragment length polymorphism (AFLP) techniques. Q-PCR analysis showed that snaB and snbA involved, respectively, in the biosynthesis of pristinamycins II and I component had more extended high expression in the recombinant than that in the ancestor during fermentation process, indicating their expression changes might be key factors during the biosynthesis of the antibiotic. In addition, the antecedent establishment of the high self-resistance to pristinamycin, because ptr resistance gene started high-level expression ahead of the onset of the antibiotic production in the recombinant, might also lead to the increase of the antibiotics yield. AFLP analysis of these recombinants revealed genome variation of two novel genes, the homologs of AfsR regulatory gene and transposase gene, indicating these two gene variations were probably responsible for yield improvement of pristinamycin. This study provided several potential molecular clues for pristinamycin yield enhancement. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0233-1 Authors Qingchao Jin, Department of Biological and Pharmaceutical Engineering, Ningbo Institute of Technology, Zhejiang University, Ningbo, China Zhihua Jin, Department of Biological and Pharmaceutical Engineering, Ningbo Institute of Technology, Zhejiang University, Ningbo, China Lijing Zhang, Department of Biological and Pharmaceutical Engineering, Ningbo Institute of Technology, Zhejiang University, Ningbo, China Shanjing Yao, Institute of Bioengineering, College of Materials Science and Chemical Engineering, Zhejiang University, Hangzhou, China Fuyong Li, Department of Internal Medicine, Weifang Municipal Hospital, Weifang, China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    While much is known about the transcriptional regulation of the 12 gene alginate biosynthesis operon from the algD promoter in Pseudomonas aeruginosa , there has been little investigation into the possibility of other transcription starts within this operon, especially those genes dealing with the epimerization and acetylation of the alginate polymer. In this study, we utilized quantitative reverse transcription polymerase chain reaction, a β-galactosidase reporter assay and sequence scanning to identify two putative promoters within the alginate biosynthesis operon upstream of the alginate epimerase gene algG and the alginate acetylation gene algI . These data support the possibility of differential regulation within the operon to alter polymer structure under varying environmental conditions. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0228-y Authors Janice L. Paletta, Department of Microbiology and Immunology, Virginia Commonwealth University Medical Center, P.O. Box 980678, Richmond, VA 23298-0678, USA Dennis E. Ohman, Department of Microbiology and Immunology, Virginia Commonwealth University Medical Center, P.O. Box 980678, Richmond, VA 23298-0678, USA Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Helicobacter pylori able to express green fluorescent protein, as well as an ATCC strain, and a clinical isolate of this pathogen were evaluated for their ability to survive predation by Acanthamoeba polyphaga . Ingestion was evaluated by microscopic observation of the GFP- H. pylori and BacLight™-stained cells. Following phagocytosis, the fate of cells was assessed by fluorescent in situ hybridization with an oligonucleotide targeting H. pylori 16S rRNA and by quantitative-polymerase chain reaction (qPCR) tests with primers to 16S rDNA. Fluorescent in situ hybridization tests were inconclusive with only a small percentage of amoebae apparently containing active intracellular H. pylori . Furthermore, no increase in bacterial cells was detected by qPCR. Additional research is required to elucidate the mechanisms by which amoebae phagocytize this important bacterial pathogen. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-012-0232-2 Authors Charlotte D. Smith, Charlotte Smith & Associates, Inc., PO Box 629, Orinda, CA 94563, USA Nicholas J. Ashbolt, National Exposure Research Laboratory (MD-593), U. S. Environmental Protection Agency, 26 W. Martin Luther King Drive, Cincinnati, OH 45268, USA Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Wireworms, the polyphagous larvae of click beetles belonging to the genus Agriotes (Coleoptera: Elateridae) are severe and widespread agricultural pests that affect numerous crops globally. A new bacterial specimen identified in diseased wireworms had previously been shown by microscopy and 16S ribosomal RNA (rRNA) gene-based phylogenetic reconstruction to belong to the taxonomic genus Rickettsiella ( Gammaproteobacteria ) that comprises intracellular bacteria associated with and typically pathogenic for a wide range of arthropods. Going beyond these earlier results obtained from rRNA phylogenies, multilocus sequence analysis (MLSA) using a four marker scheme has been employed in the molecular taxonomic characterization of the new Rickettsiella pathotype, referred to as ‘ Rickettsiella agriotidis ’. In combination with likelihood-based significance testing, the MLSA approach demonstrated the close phylogenetic relationship of ‘ R. agriotidis ’ to the pathotypes ‘ Rickettsiella melolonthae ’ and ‘ Rickettsiella tipulae ’, i.e., subjective synonyms of the nomenclatural type species, Rickettsiella popilliae . ‘ R. agriotidis ’ forms, therefore, part of a Rickettsiella pathotype complex that most likely represents the species R. popilliae . As there are currently no genetic data available from the R. popilliae type strain, the respective assignment cannot be corroborated directly. However, an alternative taxonomic assignment to the species Rickettsiella grylli has been positively ruled out by significance testing. MLSA has been shown to provide a more powerful tool for taxonomic delineation within the genus Rickettsiella as compared to 16S rRNA phylogenetics. However, the limitations of the present MLSA scheme for the sub-species level classification of ‘ R. agriotidis ’ and further R. popilliae synonyms has been critically evaluated. Content Type Journal Article Pages 1-9 DOI 10.1007/s00284-012-0219-z Authors Christina Schuster, Institut für Biologischen Pflanzenschutz, Julius Kühn-Institut (JKI), Heinrichstraße 243, 64287 Darmstadt, Germany Regina G. Kleespies, Institut für Biologischen Pflanzenschutz, Julius Kühn-Institut (JKI), Heinrichstraße 243, 64287 Darmstadt, Germany Claudia Ritter, Centre for Field Vegetable Production (GKZ), State Institute for Agriculture and Fishing Research Mecklenburg-West Pomerania (LFA), Dorfplatz 1, 18276 Gülzow, Germany Simon Feiertag, Institut für Biologischen Pflanzenschutz, Julius Kühn-Institut (JKI), Heinrichstraße 243, 64287 Darmstadt, Germany Andreas Leclerque, Institut für Biologischen Pflanzenschutz, Julius Kühn-Institut (JKI), Heinrichstraße 243, 64287 Darmstadt, Germany Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Bacteria acquire new DNA in a process known as horizontal gene transfer (HGT). To investigate the evolutionary impact of this transfer of DNA, various methods have been developed to detect past HGT events. For example, codon usage-based methods detect the presence of transferred genes by identifying atypical patterns of codon usage. However, some inherited genes exhibit atypical codon usage and some transferred genes have codon usage patterns similar to those of the inherited genes. In this study, we used a comparative phylogenetic approach with Methylobacterium and Caulobacter species to demonstrate that even well-designed codon usage methods fail to detect many HGT events and generate a high rate of false positives (60–75 %) and false negatives (23–61 %). Therefore, we recommend caution when employing codon usage methods to identify transferred genes and suggest that the rapidly increasing availability of bacterial genome sequences makes the phylogenetic approach the method of choice. Content Type Journal Article Pages 639-642 DOI 10.1007/s00284-012-0205-5 Authors Robert Friedman, Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA Bert Ely, Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651 Journal Volume Volume 65 Journal Issue Volume 65, Number 5

Description:    The distribution and diversity of acyl homoserine lactone (AHL) producing bacteria in black, brown, red, and meadow soils were investigated using culture-dependent method. One out of seventy six and five out of fifty isolates from the black and the brown soil were AHL-biosensor active, respectively. They were affiliated to the genera Ensifer and Pseudomonas and the family Enterobacteriaceae . Thin layer chromatography showed that the most of them produced AHLs with acyl side chain of 6 or 8 carbons. No AHL-producer was found in red and meadow soils. A potential novel AHL-based quorum sensing and quenching system was identified by sequencing in the black soil isolate Ensifer adhaerens X097 that harbored two AHL synthetase-like proteins and one amidohydrolase-like protein. This is the first report of comparison of AHL-producers among different soils. Our data showed that composition of AHL-producers were niche specific and were not in proportion with community population. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0234-0 Authors Yili Huang, Zhejiang Provincial Key Laboratory of Organic Pollution Process and Control, Hangzhou, China Yanhua Zeng, Zhejiang Provincial Key Laboratory of Organic Pollution Process and Control, Hangzhou, China Zhiliang Yu, College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou, China Jing Zhang, Zhejiang Provincial Key Laboratory of Organic Pollution Process and Control, Hangzhou, China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    A Gram-negative, aerobic, rod-shaped, motile by gliding and yellow-pigmented bacterium, designated strain 6Alg 8 T , was isolated from the common Pacific green alga Ulva fenestrata . The phylogenetic analysis based on 16S rRNA gene sequence placed the novel strain within the genus Polaribacter , a member of the family Flavobacteriaceae , the phylum Bacteroidetes , with sequence similarities of 97.6 % to Polaribacter dokdonensis DSW-5 T and 92.8–96.1 % to other recognized Polaribacter species. The prevalent fatty acids of strain 6Alg 8 T were iso-C 15:0 , iso-C 15:1 , iso-C 15:0 2-OH, C 15:0 and C 15:1 ω6. The polar lipid profile consisted of the major lipids phosphatidylethanolamine, two unknown aminolipids and one unknown lipid. The DNA G+C content of the type strain is 31.6 mol%. The new isolate and the type strains of recognized species of the genus Polaribacter were readily distinguished based on a number of phenotypic characteristics. A combination of the genotypic and phenotypic data showed that the algal isolate represents a novel species of the genus Polaribacter , for which the name Polaribacter reichenbachii sp. nov. is proposed. The type strain is 6Alg 8 T (= KCTC 23969 T  = KMM 6386 T  = LMG 26443 T ). Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0200-x Authors Olga I. Nedashkovskaya, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far-Eastern Branch of the Russian Academy of Sciences, Pr. 100 Let Vladivostoku 159, 690022 Vladivostok, Russia Andrey D. Kukhlevskiy, A.V. Zhirmunsky Institute of Marine Biology, Far-Eastern Branch of the Russian Academy of Sciences, Pal’chevskogo St. 17, 690032 Vladivostok, Russia Natalia V. Zhukova, A.V. Zhirmunsky Institute of Marine Biology, Far-Eastern Branch of the Russian Academy of Sciences, Pal’chevskogo St. 17, 690032 Vladivostok, Russia Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Leptospirosis is a globally prevalent zoonosis caused by pathogenic Leptospira spp.; several serologic variants have reservoirs in synanthropic rodents. The capybara is the largest living rodent in the world, and it has a wide geographical distribution in Central and South America. This rodent is a significant source of Leptospira since the agent is shed via urine into the environment and is a potential public health threat. In this study, we isolated and identified by molecular techniques a pathogenic Leptospira from capybara in southern Brazil. The isolated strain was characterized by partial rpoB gene sequencing and variable-number tandem-repeats analysis as L. interrogans , serogroup Icterohaemorrhagiae. In addition, to confirm the expression of virulence factors, the bacterial immunoglobulin-like proteins A and B expression was detected by indirect immunofluorescence using leptospiral specific monoclonal antibodies. This report identifies capybaras as an important source of infection and provides insight into the epidemiology of leptospirosis. Content Type Journal Article Pages 1-4 DOI 10.1007/s00284-012-0169-5 Authors Sérgio Jorge, Laboratório de Vacinologia, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Brazil Leonardo G. Monte, Laboratório de Imunodiagnóstico, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Brazil Marco Antonio Coimbra, Núcleo de Reabilitação da Fauna Silvestre, Instituto de Biologia, Universidade Federal de Pelotas, Pelotas, Brazil Ana Paula Albano, Núcleo de Reabilitação da Fauna Silvestre, Instituto de Biologia, Universidade Federal de Pelotas, Pelotas, Brazil Daiane D. Hartwig, Laboratório de Vacinologia, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Brazil Caroline Lucas, Laboratório de Genômica Funcional, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Brazil Fabiana K. Seixas, Laboratório de Genômica Funcional, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Brazil Odir A. Dellagostin, Laboratório de Vacinologia, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Brazil Cláudia P. Hartleben, Laboratório de Imunodiagnóstico, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Brazil Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    In Pseudomonas aeruginosa , quorum sensing (QS) autoinducer known as acyl homoserine lactone (AHL) acts as a key regulator in the expression of pathogenic characters. In this work, the efficiency of phenylacetic acid (PAA) in reducing the production of AHL-dependent factors in P. aeruginosa PAO1 was studied. PAA at a concentration of 200 μg ml −1 displayed significant reduction in QS-dependent pyocyanin, exopolysaccharide, and protease and elastase production in PAO1. In swimming inhibition assay, PAA-treated PAO1 cells exhibited poor motility in swimming agar plate. In in vivo analysis, PAO1-preinfected Caenorhabditis elegans showed enhanced survival when treated with PAA. PAA at the QS inhibitory concentration showed no growth inhibitory activity on PAO1. Results of the present study revealed the potential of PAA as antipathogenic compound to prevent QS-dependent pathogenicity of P. aeruginosa . Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0181-9 Authors Khadar Syed Musthafa, Department of Biotechnology, Alagappa University, Karaikudi, 630 003 Tamil Nadu, India Bhagavathi Sundaram Sivamaruthi, Department of Biotechnology, Alagappa University, Karaikudi, 630 003 Tamil Nadu, India Shunmugiah Karutha Pandian, Department of Biotechnology, Alagappa University, Karaikudi, 630 003 Tamil Nadu, India Arumugam Veera Ravi, Department of Biotechnology, Alagappa University, Karaikudi, 630 003 Tamil Nadu, India Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description: Characterization of Plasmid pML21 of Enterococcus faecalis ML21 from Koumiss Content Type Journal Article Pages 1-3 DOI 10.1007/s00284-012-0255-8 Authors Fanglei Zuo, Key Laboratory of Functional Dairy Science of Chinese Ministry of Education and Municipal Government of Beijing, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083 People’s Republic of China Xiujuan Feng, Key Laboratory of Functional Dairy Science of Chinese Ministry of Education and Municipal Government of Beijing, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083 People’s Republic of China Xiaofei Sun, Key Laboratory of Functional Dairy Science of Chinese Ministry of Education and Municipal Government of Beijing, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083 People’s Republic of China Chao Du, Key Laboratory of Functional Dairy Science of Chinese Ministry of Education and Municipal Government of Beijing, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083 People’s Republic of China Shangwu Chen, Key Laboratory of Functional Dairy Science of Chinese Ministry of Education and Municipal Government of Beijing, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083 People’s Republic of China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    In Pseudomonas aeruginosa PAO1, the pvdQ gene has been shown to have at least two functions. It encodes the acylase enzyme and hydrolyzes 3-oxo-C12-HSL, the key signaling molecule of quorum sensing system. In addition, pvdQ is involved in swarming motility. It is required and up-regulated during swarming motility, which is triggered by high cell densities. As high density bacterial populations also display elevated antibiotics resistance, studies have demonstrated swarm-cell differentiation in P. aeruginosa promotes increased resistance to various antibiotics. PvdQ acts as a signal during swarm-cell differentiation, and thus may play a role in P. aeruginosa antibiotic resistance. The aim of this study was to examine whether pvdQ was involved in modifying antibiotic susceptibility during swarming conditions and to investigate the mechanism by which this occurred. We constructed the PAO1pME pvdQ strain, which overproduces PvdQ. PAO1pME pvdQ promotes swarming motility, while PAO1Δ pvdQ abolishes swarming motility. In addition, both PAO1 and PAO1pME pvdQ acquired resistance to ceftazidime, ciprofloxacin, meropenem, polymyxin B, and gentamicin, though PAO1pME pvdQ exhibited a twofold to eightfold increase in antibiotic resistance compared to PAO1. These results indicate that pvdQ plays an important role in elevating antibiotic resistance via swarm-cell differentiation and possibly other mechanisms as well. We analyzed outer membrane permeability. Our data also suggest that pvdQ decreases P. aeruginosa outer membrane permeability, thereby elevating antibiotic resistance under swarming conditions. Our results suggest new approaches for reducing P. aeruginosa resistance. Content Type Journal Article Pages 1-10 DOI 10.1007/s00284-012-0217-1 Authors Lili Wang, Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030 Hubei Province, China Chunling Zhang, Department of Respiratory Medicine, Central Hospital of Qingdao, Qingdao, 266042 Shandong Province, China Fengyun Gong, Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030 Hubei Province, China Hongtao Li, Department of Oncology, Affiliated People’s 6th Hospital, Shanghai Jiaotong University, Shanghai, 200233 China Xuhua Xie, Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030 Hubei Province, China Chao Xia, Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030 Hubei Province, China Jia Chen, Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030 Hubei Province, China Ying Song, Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030 Hubei Province, China Aixia Shen, Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030 Hubei Province, China Jianxin Song, Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030 Hubei Province, China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The bacteria Xenorhabdus spp. are entomopathogenic symbionts that can produce several toxic proteins that interfere the immune system of insects. We purified an insecticidal protein from Xenorhabdus ehlersii , and designated it as XeGroEL with an estimated molecular mass of ~58 kDa. Galleria mellonella larva injected with XeGroEL presented prophenoloxidase activation and hemocyte decrease. XeGroEL can kill G. mellonella larva in 48 h with an LD 50 of 0.76 ± 0.08 μg/larva. Our results demonstrate that X. ehlersii possesses a toxic XeGroEL protein acting as a potential factor to activate proPO in host insect, which also provides a meaningful hypothesis to understand the interaction between nematode-symbiotic bacteria and host. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0114-7 Authors Huaixing Shi, Key Laboratory of Integrated Pest Management in Crops, Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081 China Hongmei Zeng, Key Laboratory of Integrated Pest Management in Crops, Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081 China Xiufen Yang, Key Laboratory of Integrated Pest Management in Crops, Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081 China Jing Zhao, Key Laboratory of Integrated Pest Management in Crops, Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081 China Mingjia Chen, Key Laboratory of Integrated Pest Management in Crops, Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081 China Dewen Qiu, Key Laboratory of Integrated Pest Management in Crops, Ministry of Agriculture, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081 China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Anaerobic gram-negative oral bacteria such as Treponema denticola , Aggregatibacter actinomycetemcomitans , Porphyromonas gingivalis , Tannerella forsythia , Campylobacter rectus , and Fusobacterium nucleatum are closely associated with periodontal diseases. We measured the relative population (bacterial levels) of these oral pathogens in subgingival tissues of patients at different stages of Korean chronic periodontal diseases. We divided the individuals into those with chronic gingivitis (G), moderate periodontitis (P1), severe periodontitis (P2), and normal individuals (N) ( n  = 20 for each group) and subgingival tissue samples were collected. We used real-time PCR with TaqMan probes to evaluate the change of periodontal pathogens among different stages of periodontitis. Bacterial levels of A. actinomycetemcomitans and C. rectus are significantly increased in individuals with chronic gingivitis and moderate periodontitis, but unchanged in severe periodontitis patients. These results suggest that analyzing certain bacterial levels among total oral pathogens may facilitate understanding of the role of periodontal bacteria in the early stages of periodontitis. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0121-8 Authors Heon-Jin Lee, Department of Oral Microbiology, School of Dentistry, Kyungpook National University, 2-188-1 Samduk-dong, Jung-gu, Daegu, 700-412 South Korea Jin-Kyoung Kim, Department of Oral Microbiology, School of Dentistry, Kyungpook National University, 2-188-1 Samduk-dong, Jung-gu, Daegu, 700-412 South Korea Je-Yeol Cho, Department of Oral Biochemistry, School of Dentistry, Kyungpook National University, 2-188-1 Samduk-dong, Jung-gu, Daegu, 700-412 South Korea Jae-Mok Lee, Department of Periodontology, School of Dentistry, Kyungpook National University, 2-188-1 Samduk-dong, Jung-gu, Daegu, 700-412 South Korea Su-Hyung Hong, Department of Oral Microbiology, School of Dentistry, Kyungpook National University, 2-188-1 Samduk-dong, Jung-gu, Daegu, 700-412 South Korea Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Six structurally related 3-keto-substituted macrolide antibiotics (ketolides) were compared for concentration-dependent inhibitory effects on growth rate, viable cell number, and protein synthesis rates in Staphylococcus aureus cells. Inhibitory effects on 50S ribosomal subunit formation were also examined, as this is a second target for these antibiotics. A concentration range of 0.01 to 0.1 μg/ml was tested. An IC 50 for inhibition of translation and 50S synthesis was measured for each compound, to relate structural features to inhibitory activity. ABT-773 was the most effective of the six compounds tested with an IC 50 = 0.035 μg/ml. HMR 3004 was almost as effective with an IC 50 = 0.05 μg/ml. Two 2-fluoroketolides (HMR 3562 and HMR 3787) were equivalent in their inhibitory activity with an IC 50 = 0.06 μg/ml. Telithromycin (HMR 3647) had an IC 50 = 0.08 μg/ml, and HMR 3832 was least effective with an IC 50 = 0.11 μg/ml. Each antibiotic had an equivalent inhibitory effect on translation and 50S subunit formation. These results indicate specific structural features of these antimicrobial agents, which contribute to defined inhibitory activities against susceptible organisms. Content Type Journal Article Pages 203-210 DOI 10.1007/PL00021055 Authors W. Scott Champney, Department of Biochemistry and Molecular Biology, J.H. Quillen College of Medicine, East Tennessee State University,, Johnson City, TN 37614, USA, US Craig L. Tober, Department of Biochemistry and Molecular Biology, J.H. Quillen College of Medicine, East Tennessee State University,, Johnson City, TN 37614, USA, US Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651 Journal Volume Volume 42 Journal Issue Volume 42, Number 3

Description:    Human liver was closely associated with gut through various biological mechanisms, such as bacterium–gut interactions. Alterations of gut microbiota seemed to play an important role in induction and promotion of liver damage progression. The aim of this study was to characterize the gut microbiota in liver cirrhosis patients and assess whether there are alterations in the diversity and similarity of intestinal flora in cirrhotic patients when compared with healthy individuals. PCR-denaturing gradient gel electrophoresis (DGGE) with universal primers targeting V3 region of the 16S rRNA gene was employed to characterize the overall intestinal microbiota composition, and some excised gel bands were cloned for sequencing. Real-time PCR was further utilized to quantitatively analyze the subpopulation of microbiota using group-specific primers targeting the Enterobacteriaceae , Enterococcus and Bifidobacterium genus. The DGGE profiles of two groups demonstrated significant differences between cirrhotic and healthy groups ( P  〈 0.05). While real-time PCR revealed significant increase of Enterobacteriaceae and Enterococcus ( P  〈 0.05) in the cirrhotic group compared with the healthy group. The ratio of Bifidobacterium genus and Enterobacteriaceae decreased in the cirrhotic patients group, but no statistical significance. This study revealed strong relationship between alterations of gut microbiota and liver cirrhosis. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0105-8 Authors Jianjun Liu, Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian, China Dachang Wu, Department of Biotechnology, Dalian Medical University, Dalian, 116044 China Ayaz Ahmed, Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian, China Xinli Li, Department of Biotechnology, Dalian Medical University, Dalian, 116044 China Yufang Ma, Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian, China Li Tang, Department of Microecology, Dalian Medical University, Dalian, China Dianjun Mo, Department of Laboratory Medicine, The Affiliated Hospital of Chifeng Medical University, Chifeng, China Yue Ma, Department of Laboratory Medicine, The Second Affiliated Hospital of Dalian Medical University, Dalian, China Yi Xin, Department of Biotechnology, Dalian Medical University, Dalian, 116044 China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Mercury pollution is a major environmental problem that arises as a result of natural processes as well as from anthropogenic sources. In response to toxic mercury compounds, microbes have developed astonishing array of resistance systems to detoxify them. To address this challenge, this study was aimed in screening bacterial isolates for their tolerance against varied concentrations of phenylmercuric acetate. Mercury transformation by bacteria being sensitive to factors such as available carbon source, etc. that affect mer -mediated transformation, screened mercury tolerant bacteria were also studied for their tolerance to different antimicrobials and carbon sources, followed by identification using biochemical as well as 16S rRNA approach. Following identification, gene encoding organomercurial lyase catalyzing protonolytic cleavage of C–Hg bond of organic mercury was amplified using gene specific primers, cloned in pGEMT ® easy vector and sequenced. Microbe-based approach using organomercurial lyase encoded by merB gene being potentially economic, provides foundation to facilitate genetic manipulation of this environmentally important enzyme to remove high concentrations of obstinate mercury using holistic, multifaceted approach for use in bioremediation through generation of transgenics or as catalyst for use in bioreactors. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-012-0118-3 Authors Arif Tasleem Jan, Department of Biosciences, Jamia Millia Islamia, New Delhi, 110025 India Mudsser Azam, Department of Biosciences, Jamia Millia Islamia, New Delhi, 110025 India Arif Ali, Department of Biosciences, Jamia Millia Islamia, New Delhi, 110025 India Qazi Mohd. Rizwanul Haq, Department of Biosciences, Jamia Millia Islamia, New Delhi, 110025 India Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The contribution of RecA, Dps, and RpoS to survival of Escherichia coli O157:H7 during desiccation and osmotic stress was determined in Luria–Bertani broth with 12 % NaCl (LB-12) at 30 and 37 °C, on filter disks at 23 and 30 °C, and in sterile bovine feces at 30 °C. RecA did not significantly contribute to survival in any condition or temperature. The contribution of Dps to survival was only significant in LB-12 at 37 °C. RpoS was necessary for survival during desiccation and osmotic stress, and survival of the RpoS mutant was significantly less than the parent in all conditions and temperatures. The RpoS mutant survived up to 21 days in bovine feces, 〈4 days on filter disks, and 〉8 and 〈4 days in LB-12 at 30 and 37 °C, respectively. The parent, ΔrecA , dps , and dps / ΔrecA mutant strains survived 〉8 days in LB-12, 〉28 days on filter disks, and 〉28 days in bovine feces. Increased incubation temperatures were associated with decreased survival. E. coli O157:H7 can persist in desiccating and osmotically challenging environments, especially sterile feces, for an extended period time. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0210-8 Authors Andrew J. Stasic, Department of Bacteriology, Food Research Institute, University of Wisconsin-Madison, 1550 Linden Drive, Madison, WI 53706, USA Amy C. Lee Wong, Department of Bacteriology, Food Research Institute, University of Wisconsin-Madison, 1550 Linden Drive, Madison, WI 53706, USA Charles W. Kaspar, Department of Bacteriology, Food Research Institute, University of Wisconsin-Madison, 1550 Linden Drive, Madison, WI 53706, USA Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Unlike dairy lactic acid bacteria, Lactobacillus brevis cannot ferment milk. We characterized the lactose utilization by L. brevis KB290. In a carbohydrate fermentation assay using API 50 CHL, we showed during 7 days L. brevis did not ferment lactose. L. brevis grew to the stationary phase in 2 weeks in MRS broth containing lactose as the carbon source. L. brevis slowly consumed the lactose in the medium. L. brevis hydrolyzed lactose and a lactose analog, o -nitrophenyl-β- d -galactopyranoside (ONPGal). This β-galactosidase activity for ONPGal was not repressed by glucose, galactose, fructose, xylose, or maltose showing the microorganism may not have carbon catabolite repression. We purified the L. brevis β-galactosidase using ammonium sulfate precipitation and several chromatographies. The enzyme’s molecular weight is estimated at 72 and 37 kDa using SDS-PAGE analysis. The N-terminal amino acid sequence of the larger protein was 90 % similar to the sequence of the putative β-galactosidase (YP_796339) and the smaller protein was identical to the sequence of the putative β-galactosidase (YP_796338) in L. brevis ATCC367. This suggests the enzyme is a heterodimeric β-galactosidase. The specific activity of the purified enzyme for lactose is 55 U/mg. We speculate inhibition of lactose transport delays the lactose utilization in L. brevis KB290. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0216-2 Authors Hiroyuki Honda, Research Institute, Kagome Co., Ltd., 17 Nishitomiyama, Nasushiobara, 329-2762 Japan Nobuhiro Yajima, Research Institute, Kagome Co., Ltd., 17 Nishitomiyama, Nasushiobara, 329-2762 Japan Tadao Saito, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba, Sendai, 981-8555 Japan Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Aspergillus niger LOCK 62 produces an antifungal chitinase. Different sources of chitin in the medium were used to test the production of the chitinase. Chitinase production was most effective when colloidal chitin and shrimp shell were used as substrates. The optimum incubation period for chitinase production by Aspergillus niger LOCK 62 was 6 days. The chitinase was purified from the culture medium by fractionation with ammonium sulfate and affinity chromatography. The molecular mass of the purified enzyme was 43 kDa. The highest activity was obtained at 40 °C for both crude and purified enzymes. The crude chitinase activity was stable during 180 min incubation at 40 °C, but purified chitinase lost about 25 % of its activity under these conditions. Optimal pH for chitinase activity was pH 6–6.5. The activity of crude and purified enzyme was stabilized by Mg 2+ and Ca 2+ ions, but inhibited by Hg 2+ and Pb 2+ ions. Chitinase isolated from Aspergillus niger LOCK 62 inhibited the growth of the fungal phytopathogens: Fusarium culmorum , Fusarium solani and Rhizoctonia solani . The growth of Botrytis cinerea , Alternaria alternata , and Fusarium oxysporum was not affected. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0208-2 Authors Maria Swiontek Brzezinska, Department of Environmental Microbiology and Biotechnology, Institute of Ecology and Environmental Protection, Nicolaus Copernicus University, Gagarina 9, Toruń, Poland Urszula Jankiewicz, Department of Biochemistry, Warsaw University of Life Sciences, Nowoursynowska 159, Warsaw, Poland Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Wood-feeding termites live on cellulolytic materials that typically lack of nitrogen sources. It was reported that symbiotic microbes play important roles in the maintenance of a normal nitrogen contents in termite by different metabolisms including nitrogen fixation. In this study, the diversity of nitrogen-fixing organisms in the symbiotic intestinal microflora of Reticulitermes chinensis Snyder was investigated with culture independent method. Fragments of the nifH genes, which encode dinitrogenase reductase, were directly amplified from the DNA of the mixed microbial population in the termite gut with four sets of primers corresponding to the conserved regions of the genes. Clones were randomly selected and analyzed by RFLP. Sequence analysis revealed that a large number of nifH sequences retrieved from the termite gut were most closely related to strict anaerobic bacteria such as clostridia and spirochetes, some of the others were affiliated with proteobacteria, bacteroides, or methanogenic archaea. The results showed that there was a remarkable diversity of nitrogenase genes in the gut of Reticulitermes chinensis Snyder. Content Type Journal Article Pages 1-5 DOI 10.1007/s00284-012-0185-5 Authors Xin Du, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Sciences, Central China Normal University, 152 Luoyu Avenue, Wuhan, 430079 People’s Republic of China Xiaojuan Li, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Sciences, Central China Normal University, 152 Luoyu Avenue, Wuhan, 430079 People’s Republic of China Yin Wang, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Sciences, Central China Normal University, 152 Luoyu Avenue, Wuhan, 430079 People’s Republic of China Jianxin Peng, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Sciences, Central China Normal University, 152 Luoyu Avenue, Wuhan, 430079 People’s Republic of China Huazhu Hong, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Sciences, Central China Normal University, 152 Luoyu Avenue, Wuhan, 430079 People’s Republic of China Hong Yang, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Sciences, Central China Normal University, 152 Luoyu Avenue, Wuhan, 430079 People’s Republic of China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    In the last few years, numerous cases of multidrug-resistant Achromobacter xylosoxidans infections have been documented in immunocompromised and cystic fibrosis patients. To gain insights into the molecular mechanisms and mobile elements related to multidrug resistance in this bacterium, we studied 24 non-epidemiological A. xylosoxidans clinical isolates from Argentina. Specific primers for plasmids, transposons, insertion sequences, bla ampC , intI1 , and intI2 genes were used in PCR reactions. The obtained results showed the presence of wide host range IncP plasmids in ten isolates and a high dispersion of class 1 integrons ( n  = 10) and class 2 integrons ( n  = 3). Four arrays in the variable region (vr) of class 1 integrons were identified carrying different gene cassettes as the aminoglycoside resistance aac(6′) - Ib and aadA1 , the trimethoprim resistance dfrA1 and dfrA16 , and the β-lactamase bla OXA-2 . In only one of the class 2 integrons, a vr was amplified that includes sat2 - aadA1 . The bla ampC gene was found in all isolates, confirming its ubiquitous nature. Our results show that A. xylosoxidans clinical isolates contain a rich variety of genetic elements commonly associated with resistance genes and their dissemination. This supports the hypothesis that A. xylosoxidans is becoming a reservoir of horizontal genetic transfer elements commonly involved in spreading antibiotic resistance. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0213-5 Authors German Matías Traglia, Laboratorio de Investigaciones de los Mecanismos de Resistencia a Antibióticos, Instituto de Microbiología y Parasitología Médica (IMPaM, UBA-CONICET), Facultad de Medicina, Universidad de Buenos Aires, CONICET, Paraguay 2155 Piso 12, 1121 Buenos Aires, Argentina Marisa Almuzara, Laboratorio de Bacteriología Clínica, Departamento de Bioquímica Clínica, Instituto de Fisiopatología y Bioquímica Clínica, Hospital de Clínicas José de San Martín, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina Andrea Karina Merkier, Laboratorio de Investigaciones de los Mecanismos de Resistencia a Antibióticos, Instituto de Microbiología y Parasitología Médica (IMPaM, UBA-CONICET), Facultad de Medicina, Universidad de Buenos Aires, CONICET, Paraguay 2155 Piso 12, 1121 Buenos Aires, Argentina Christina Adams, Laboratorio de Investigaciones de los Mecanismos de Resistencia a Antibióticos, Instituto de Microbiología y Parasitología Médica (IMPaM, UBA-CONICET), Facultad de Medicina, Universidad de Buenos Aires, CONICET, Paraguay 2155 Piso 12, 1121 Buenos Aires, Argentina Laura Galanternik, Hospital de Niños Ricardo Gutiérrez, Buenos Aires, Argentina Carlos Vay, Laboratorio de Bacteriología Clínica, Departamento de Bioquímica Clínica, Instituto de Fisiopatología y Bioquímica Clínica, Hospital de Clínicas José de San Martín, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina Daniela Centrón, Laboratorio de Investigaciones de los Mecanismos de Resistencia a Antibióticos, Instituto de Microbiología y Parasitología Médica (IMPaM, UBA-CONICET), Facultad de Medicina, Universidad de Buenos Aires, CONICET, Paraguay 2155 Piso 12, 1121 Buenos Aires, Argentina María Soledad Ramírez, Laboratorio de Investigaciones de los Mecanismos de Resistencia a Antibióticos, Instituto de Microbiología y Parasitología Médica (IMPaM, UBA-CONICET), Facultad de Medicina, Universidad de Buenos Aires, CONICET, Paraguay 2155 Piso 12, 1121 Buenos Aires, Argentina Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The antimicrobial properties of methyl (MTS) and ethyl (ETS) esters of thiosulfonic acid alone and in combination with rhamnolipid-biosurfactant (RL) have been characterized for their ability to disrupt the normal physiological functions of living pathogens. Bactericidal and fungicidal activities of MTS and ETS and their combination with rhamnolipid were demonstrated on strains of Pseudomonas aeruginosa, Bacillus subtilis, Alcaligenes faecalis, and Rhizopus ngtricans . It was found that the combination of rhamnolipid and thiosulfonic esters has a synergistic effect leading to decreasing of bactericidal and fungicidal concentrations of MTS and ETS. More extensively was studied the effect of rhamnolipid on the lipid composition of B. subtilis bacterial membrane. To our knowledge, in this article is reported for the first time a remarkable increase of negatively charged phospholipid cardiolipin in the presence of rhamnolipid. The capacity of RL as a surface-active substance was confirmed by scanning electron microscopy (SEM). The occurrence of surface infolds and blebs on B. subtilis shown by SEM, was not accompanied by changes in membrane permeability tested by a live/dead viability staining for fluorescence microscopy. When RL was applied in combination with MTS, a dramatic permeability shift for propidium iodide was observed in vegetative cells. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-012-0191-7 Authors Anna Sotirova, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Acad. G. Bonchev St., Block 26, 1113 Sofia, Bulgaria Tatyana Avramova, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Acad. G. Bonchev St., Block 26, 1113 Sofia, Bulgaria Stoyanka Stoitsova, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Acad. G. Bonchev St., Block 26, 1113 Sofia, Bulgaria Irina Lazarkevich, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Acad. G. Bonchev St., Block 26, 1113 Sofia, Bulgaria Vera Lubenets, Institute of Physical Chemistry, Ukrainian Academy of Sciences, Bandera Str. 12, Lviv, Ukraine Elena Karpenko, Institute of Physical Chemistry, Ukrainian Academy of Sciences, Bandera Str. 12, Lviv, Ukraine Danka Galabova, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Acad. G. Bonchev St., Block 26, 1113 Sofia, Bulgaria Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The identification of Trichoderma genes whose expression is altered during early stages of interaction with developing roots of germinated seeds is an important step toward understanding the rhizosphere competency of Trichoderma spp. The potential of 13 Trichoderma strains to colonize tomato root and promote plant growth has been evaluated. All used strains successfully propagated in spermosphere and continued their growth in rhizoplane simultaneously root enlargement while the strains T6 and T7 were the most abundant in the apical segment of roots. Root colonization in most strains associated with promoting the roots and shoots growth while they significantly increased up to 43 and 40 % roots and shoots dry weights, respectively. Differential display reverse transcriptase-PCR (DDRT-PCR) has been developed to detect differentially expressed genes in the previously selected strain, Trichoderma harzianum T7, during colonization stages of tomato-germinating seeds and roots. Amplified DDRT-PCR products were analyzed on gel agarose and 62 differential bands excised, purified, cloned, and sequenced. Obtained ESTs were submit-queried to NCBI database by BLASTx search and gene ontology hierarchy. Most of transcripts (29 EST) corresponds to known and hypothetical proteins such as secretion-related small GTPase, 40S ribosomal protein S3a, 3-hydroxybutyryl-CoA dehydrogenase, DNA repair protein rad50, lipid phosphate phosphatase-related protein type 3, nuclear essential protein, phospholipase A2, fatty acid desaturase, nuclear pore complex subunit Nup133, ubiquitin-activating enzyme, and 60S ribosomal protein L40. Also, 13 of these sequences showed no homology ( E  〉 0.05) with public databases and considered as novel genes. Some of these ESTs corresponded to genes encodes enzymes potentially involved in nutritional support of microorganisms which have obvious importance in the establishment of Trichoderma in spermosphere and rhizosphere, via potentially functioning in acquisition of nutrients from energy-rich carbon compounds leaked from the germinating seeds and roots. Content Type Journal Article Pages 1-10 DOI 10.1007/s00284-012-0189-1 Authors Mehdi Mehrabi-Koushki, Department of Plant Protection, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran Hamid Rouhani, Department of Plant Protection, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran Esmat Mahdikhani-Moghaddam, Department of Plant Protection, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    This study describe the use of a combination of two recently proposed typing approaches, multiple amplification of prophage locus typing (MAPLT) and multiple-locus variable-number tandem-repeat analysis (MLVA) for subdividing within Salmonella enterica serovar Heidelberg ( S. Heidelberg). The combined typing method was compared with pulsed-field gel electrophoresis (PFGE) by Simpson’s index of diversity (DI). PFGE was shown to have a DI = 0.84 and was poor at differentiation of the predominant PT1 (Phage Type 1) phenotype. In comparison, the combined MAPLT/MLVA method comprising 3 MLVA and 9 MAPLT primer pairs provided a higher differentiating ability DI = 0.92. More importantly, the combined methodology was found to be superior in the differentiation of the predominant PT1 isolates. In conclusion, this study demonstrated the potential of the rapid and simple amalgamated MAPLT/MLVA approach in determining transmission of isolates of clonal phage type groups from various environmental sources to humans. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0179-3 Authors Chun-Chun Young, School of Molecular and Biomedical Science, University of Adelaide, North Terrace, Adelaide, SA 5005, Australia Ian L. Ross, Public Health Unit, Department of Microbiology and Infectious Diseases, SA Pathology (at Women’s and Children’s Hospital), 72 King William Road, North Adelaide, SA 5006, Australia Michael W. Heuzenroeder, School of Molecular and Biomedical Science, University of Adelaide, North Terrace, Adelaide, SA 5005, Australia Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Rapid and accurate identification of mycobacteria isolates from primary culture is important due to timely and appropriate antibiotic therapy. Conventional methods for identification of Mycobacterium species based on biochemical tests needs several weeks and may remain inconclusive. In this study, a novel multiplex real-time PCR was developed for rapid identification of Mycobacterium genus, Mycobacterium tuberculosis complex (MTC) and the most common non-tuberculosis mycobacteria species including M. abscessus , M. fortuitum , M. avium complex, M. kansasii , and the M. gordonae in three reaction tubes but under same PCR condition. Genetic targets for primer designing included the 16S rDNA gene, the dnaJ gene, the gyrB gene and internal transcribed spacer (ITS). Multiplex real-time PCR was setup with reference Mycobacterium strains and was subsequently tested with 66 clinical isolates. Results of multiplex real-time PCR were analyzed with melting curves and melting temperature ( T m ) of Mycobacterium genus, MTC, and each of non-tuberculosis Mycobacterium species were determined. Multiplex real-time PCR results were compared with amplification and sequencing of 16S-23S rDNA ITS for identification of Mycobacterium species. Sensitivity and specificity of designed primers were each 100 % for MTC, M. abscessus , M. fortuitum, M. avium complex, M. kansasii , and M. gordonae. Sensitivity and specificity of designed primer for genus Mycobacterium was 96 and 100 %, respectively. According to the obtained results, we conclude that this multiplex real-time PCR with melting curve analysis and these novel primers can be used for rapid and accurate identification of genus Mycobacterium , MTC, and the most common non-tuberculosis Mycobacterium species. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0188-2 Authors Bahram Nasr Esfahani, Department of Microbiology, Faculty of Medicine, Isfahan University of Medical Sciences, Esfahān, Iran Hadi Rezaei Yazdi, Department of Microbiology, Faculty of Medicine, Isfahan University of Medical Sciences, Esfahān, Iran Sharareh Moghim, Department of Microbiology, Faculty of Medicine, Isfahan University of Medical Sciences, Esfahān, Iran Hajieh Ghasemian Safaei, Department of Microbiology, Faculty of Medicine, Isfahan University of Medical Sciences, Esfahān, Iran Hamid Zarkesh Esfahani, Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Esfahān, Iran Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Two-hundred-and-thirty-six isolates were collected from fresh flowers, bees and bee-hives. Of these, 20 isolates preferred d -fructose as carbon source, produced lactic acid and acetic acid but trace amounts of ethanol and were classified as fructophilic. Poor growth was recorded when strains were incubated anaerobically in the presence of d -glucose as sole carbon source. Good growth was, however, recorded when d -glucose was metabolized in the presence of external electron acceptors such as fructose, pyruvate and oxygen. Nineteen of the strains were classified as Lactobacillus kunkeei and one as Lactobacillus brevis based on phenotypic characteristics, 16S rRNA sequences, recA sequences and DNA homology. This is the first description of a fructophilic strain of L. brevis . Content Type Journal Article Pages 1-9 DOI 10.1007/s00284-012-0186-4 Authors Deon P. Neveling, Department of Microbiology, University of Stellenbosch, Private Bag X1, Matieland, Stellenbosch, 7602 South Africa Akihito Endo, Department of Microbiology, University of Stellenbosch, Private Bag X1, Matieland, Stellenbosch, 7602 South Africa Leon M. T. Dicks, Department of Microbiology, University of Stellenbosch, Private Bag X1, Matieland, Stellenbosch, 7602 South Africa Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    This study sought to investigate the effect of sulfur levels on changes in the fungal community composition of arbuscular mycorrhizae (AM) at the pod-setting stage and the relationship between the amount of applied sulfur and AM fungal diversity in different soybean cultivars. The objective of the research was to determine the optimal sulfur application level for different soybean cultivars and to improve soybean yield and quality from the perspective of AM fungal diversity. Three soybean cultivars, Heinong 44, Heinong 48, and Heinong 37, were selected as study materials. In addition to 0.033 g each of N, P 2 O 5 and K 2 O per kg of soil, 0, 0.02, 0.04, or 0.06 g of elemental sulfur was applied to each kg of soil for the four treatment groups, S1, S2, S3, and S4, respectively. The AM fungal community structure was analyzed in the soil and root of different soybean cultivars using the PCR-DGGE technology. The results indicated a significant effect of sulfur on the AM fungal community structure in the roots and rhizospheric soil of different soybean cultivars. The three soybean cultivars in group S2 exhibited the highest diversity in AM fungus. Significant changes in the dominant fungal species were observed in the DGGE fingerprints of each sample, and Glomus , Funneliformis , Rhizophagus , and Claroideoglomus fungi were the dominant species of AM fungus in the roots and soil of soybean. The application of an appropriate amount of sulfur improved the diversity of AM fungi in roots and rhizospheric soil of different soybean cultivars. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0183-7 Authors Weiguang Jie, Key Laboratory of Microbiology, College of Life Sciences, Heilongjiang University, Harbin, China Baiyan Cai, Key Laboratory of Microbiology, College of Life Sciences, Heilongjiang University, Harbin, China Yong Zhang, Department of Food and Environment Engineering, Heilongjiang East University, Harbin, China Jin Li, Key Laboratory of Microbiology, College of Life Sciences, Heilongjiang University, Harbin, China Jingping Ge, Key Laboratory of Microbiology, College of Life Sciences, Heilongjiang University, Harbin, China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The ability of an antimicrobial, cationic polyethylenimine (PEI+) to induce the three known extracytoplasmic stress responses of Escherichia coli was quantified. Exposure of E . coli to PEI+ in solution revealed specific, concentration-dependent induction of the Cpx extracytoplasmic cellular stress response, ~2.0–2.5-fold at 320 μg/mL after 1.5 h without significant induction of the σ E or Bae stress responses. In comparison, exposure of E. coli to a non-antimicrobial polymer, poly(ethylene oxide) (PEO), resulted in no induction of the three stress responses. The antimicrobial small molecule vanillin, a known membrane pore-forming compound, was observed to cause specific, concentration-dependent induction of the σ E stress response, ~6-fold at 640 μg/mL after 1.5 h, without significant induction of the Cpx or Bae stress responses. The different stress response induction profiles of PEI+ and vanillin suggest that although both are antimicrobial compounds, they interact with the bacterial membrane and extracytoplasmic area by unique mechanisms. EPR studies of liposomes containing spin-labeled lipids exposed to PEI+, vanillin, and PEO reveal that PEI+ and PEO increased membrane stability, whereas vanillin was found to have no effect. Content Type Journal Article Pages 1-5 DOI 10.1007/s00284-012-0182-8 Authors Blaine A. Lander, Department of Chemistry, United States Naval Academy, 572 Holloway Rd, Annapolis, MD 21402, USA Kyle D. Checchi, Department of Chemistry, United States Naval Academy, 572 Holloway Rd, Annapolis, MD 21402, USA Stephen A. Koplin, Department of Chemistry, United States Naval Academy, 572 Holloway Rd, Annapolis, MD 21402, USA Virginia F. Smith, Department of Chemistry, United States Naval Academy, 572 Holloway Rd, Annapolis, MD 21402, USA Tammy L. Domanski, Department of Biology, Anne Arundel Community College, Arnold, MD 21012, USA Daniel D. Isaac, Department of Chemistry, United States Naval Academy, 572 Holloway Rd, Annapolis, MD 21402, USA Shirley Lin, Department of Chemistry, United States Naval Academy, 572 Holloway Rd, Annapolis, MD 21402, USA Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Wolbachia and Cardinium are maternally inherited intracellular bacteria that can manipulate the reproduction of their arthropod hosts, such as by inducing cytoplasmic incompatibility (CI). Although the reproductive alteration induced by Wolbachia or Cardinium have been well investigated, the effects of these two endosymbionts co-infecting the same host are poorly understood. We found that Tetranychus piercei McGregor is naturally infected with Wolbachia and Cardinium . We performed all possible crossing combinations using naturally infected and cured strains, and the results show that Wolbachia induced a weak level of CI, while Cardinium -infected and doubly infected males caused severe CI. Wolbachia and Cardinium could not rescue CI each other; however, Wolbachia boosted the expression of Cardinium -induced CI. Quantitative PCR results demonstrated that CI was associated with the infection density of Wolbachia and Cardinium . Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-012-0190-8 Authors Lu-Yu Zhu, Department of Entomology, Nanjing Agricultural University, Nanjing, 210095 Jiangsu, China Kai-Jun Zhang, Department of Entomology, Nanjing Agricultural University, Nanjing, 210095 Jiangsu, China Yan-Kai Zhang, Department of Entomology, Nanjing Agricultural University, Nanjing, 210095 Jiangsu, China Cheng Ge, Department of Entomology, Nanjing Agricultural University, Nanjing, 210095 Jiangsu, China Tetsuo Gotoh, Laboratory of Applied Entomology and Zoology, Faculty of Agriculture, Ibaraki University, Ami, Ibaraki 300-0393, Japan Xiao-Yue Hong, Department of Entomology, Nanjing Agricultural University, Nanjing, 210095 Jiangsu, China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Vegetative cells of an ascomycetous yeast, morphologically consistent with published descriptions of Cyniclomyces guttulatus , were observed in large numbers in the feces and stomach washes of three dogs with a recurrent medical history characterized by vomiting and diarrhea. Nucleotide sequence analysis of an approximately 600 base pair fragment of the variable D1/D2 domain of large subunit (26S) ribosomal DNA of a pure culture, isolated from a Siberian Husky, revealed 98–99 % homology to sequences deposited in the GenBank as C. guttulatus . These data represent the first observation of C. guttulatus in association with canine gastrointestinal illness in the southern hemisphere and add weight to the hypothesis that this yeast may act as an opportunistic pathogen of dogs. An extended examination of wet mounts and smears prepared from feces collected from 63 dogs with no clinical symptoms of gastrointestinal illness, identified C. gluttulatus in 14 (22.2 %) of the animals, albeit at lower numbers than in diseased dogs, indicating that this yeast species is widely distributed as a component of the normal microflora of the canine gastrointestinal tract. Content Type Journal Article Pages 1-5 DOI 10.1007/s00284-012-0184-6 Authors Gilberto Flausino, Animal Parasitology, Federal Rural University of Rio de Janeiro, Seropédica, Brazil Paulo D. S. Leal, Animal Parasitology, Federal Rural University of Rio de Janeiro, Seropédica, Brazil Douglas McIntosh, Animal Parasitology, Federal Rural University of Rio de Janeiro, Seropédica, Brazil Luciana G. Amaral, Animal Parasitology, Federal Rural University of Rio de Janeiro, Seropédica, Brazil Walter L. Teixeira Filho, Animal Parasitology, Federal Rural University of Rio de Janeiro, Seropédica, Brazil Walter Flausino, Animal Parasitology, Federal Rural University of Rio de Janeiro, Seropédica, Brazil Carlos W. G. Lopes, Animal Parasitology, Federal Rural University of Rio de Janeiro, Seropédica, Brazil Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Staphylococcus aureus is a leading cause of nosocomial infections due to its resistance to diverse antibiotics. This bacterium produces a large number of extracellular virulence factors that are closely associated with specific diseases. In this study, diverse plant flavonoids were investigated to identify a novel anti-virulence compound against two S. aureus strains. Flavone, a backbone compound of flavonoids, at subinhibitory concentration (50 μg/mL), markedly reduced the production of staphyloxanthin and α-hemolysin. This staphyloxanthin reduction rendered the S. aureus cells 100 times more vulnerable to hydrogen peroxide in the presence of flavone. In addition, flavone significantly decreased the hemolysis of human red blood by S. aureus , and the transcriptional level of α-hemolysin gene hla and a global regulator gene sae in S. aureus cells. This finding supported the usefulness of flavone as a potential antivirulence agent against antibiotic-resistant S. aureus . Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0229-x Authors Jin-Hyung Lee, School of Chemical Engineering, Yeungnam University, Gyeongsan, Republic of Korea Joo-Hyeon Park, School of Chemical Engineering, Yeungnam University, Gyeongsan, Republic of Korea Moo Hwan Cho, School of Chemical Engineering, Yeungnam University, Gyeongsan, Republic of Korea Jintae Lee, School of Chemical Engineering, Yeungnam University, Gyeongsan, Republic of Korea Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    In this study, effects of antimony Sb(V) on growth, pigments content, oxygen evolution, and photosystem II (PSII) activity of Microcystis aeruginosa were investigated. JIP-test, Q A − reoxidation kinetic test and S-state test were used in this study to study the energy distribution and electron transport in PSII. Treatment with Sb(V) at various concentrations ranging from 5 to 100 mg/l had long-term effects on growth, pigments content, and oxygen evolution of M . aeruginosa . Low concentration of Sb(V) had no significant inhibition of the biomass production and PSII activity but inhibited the pigment synthesis. Growth, pigments content, oxygen evolution, and PSII activity were seriously inhibited when treated by high concentration of Sb(V) (100 mg/l). The target sites of Sb(V) toxic effect on the PSII of M . aeruginosa were mainly on the donor side and the apparatus in the light-dependent reaction. The quantum yield for photochemistry, density of reaction centers and photosynthesis performance index decreased, whereas the dissipated energy increased. PSII activity of M . aeruginosa was promoted when exposure to 50 mg/l Sb(V) by increasing the density of active reaction centers and electron transport after Q A − . Content Type Journal Article Pages 1-9 DOI 10.1007/s00284-012-0221-5 Authors Shuzhi Wang, Graduate University of Chinese Academy of Sciences, Beijing, 100049 China Xiangliang Pan, State Key Laboratory of Desert and Oasis Ecology, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, No. 818, South Beijing Road, Urumqi, 830011 China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    This study was undertaken to determine whether cyclic AMP (cAMP) or cAMP-receptor protein (CRP) modulates the activity of the autoinducer (AI)-2-mediated quorum sensing (QS) system in response to glucose availability in Vibrio vulnificus. A mutation in crp impaired V. vulnificus growth, decreased AI-2 production, and repressed the expression of smcR encoding the master regulator SmcR (a Vibrio harveyi LuxR homolog) of the AI-2-QS system, and these changes were prevented by in trans complementation of wild-type crp . Furthermore, glucose repressed smcR expression in the presence of CRP but not in its absence. A mutation in cyaA encoding adenylate cyclase, which is required for cAMP synthesis, also impaired V. vulnificus growth and repressed smcR expression, and these changes were recovered by in trans complementation of wild-type cyaA . These results indicate that cAMP or CRP modulates the AI-2-QS system in response to glucose availability in V. vulnificus , demonstrating the presence of a connection between catabolite repression and quorum sensing in V. vulnificus . Content Type Journal Article Pages 1-10 DOI 10.1007/s00284-012-0218-0 Authors Sun-Pyo Kim, Department of Emergency Medicine, Chosun University Medical School, Gwangju, Republic of Korea Choon-Mee Kim, Research Center for Resistant Cells, Chosun University Medical School, Gwangju, Republic of Korea Sung-Heui Shin, Research Center for Resistant Cells, Chosun University Medical School, Gwangju, Republic of Korea Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Genus Deinococcus is characterized by an increased resistance toward reactive oxygen species (ROS). The chromosome of five strains belonging to this genus has been sequenced and the presence of a luxS -like gene was deduced from their genome sequences. The aim of this study was to assess if a complete QS circuit is present in Deinococcus sp. and if this QS is associated with ROS. Primers for searching luxS -like gene and the putative receptor gene, namely ai2R , were designed. AI-2 signal production was evaluated by luminescence analysis using Vibrio harveyi BB170 as reporter strain. AI-2 signal was also evaluated by competitive assays using cinnamaldehyde, ascorbic acid, and 3-mercaptopropionic acid as interfering molecules. Potassium tellurite and metronidazole were used as oxidative stressors. A luxS -like gene as well as an ai2R gene was detected in strain UDEC-P1 by PCR. Cell-free supernatant of strain UDEC-P1 culture induced luminescence in V. harveyi BB170, and this property was inhibited with the three interfering molecules. The oxidative stressors metronidazole and potassium tellurite decreased Deinococcus sp. viability, but increased luminescence of the reporter strain. The results demonstrate that both a functional luxS -like gene and a putative receptor for AI-2 signal are present in Deinococcus sp. strain UDEC-P1. This finding also suggests that a complete QS circuit is present in this genus, which could be related to oxidative stress. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0225-1 Authors G. Fernandez-Bunster, Departamento de Microbiología, Facultad de Ciencias Biológicas, Universidad de Concepción, Casilla 160-C, Concepción, Chile C. Gonzalez, Departamento de Microbiología, Facultad de Ciencias Biológicas, Universidad de Concepción, Casilla 160-C, Concepción, Chile J. Barros, Departamento de Microbiología, Facultad de Ciencias Biológicas, Universidad de Concepción, Casilla 160-C, Concepción, Chile M. Martinez, Departamento de Microbiología, Facultad de Ciencias Biológicas, Universidad de Concepción, Casilla 160-C, Concepción, Chile Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Heritable endosymbiotic bacteria associated with insects are ubiquitous and taxonomically diverse. Many of these endosymbionts influence the fitness of their hosts and/or manipulate their host reproduction. Exploiting the effects of endosymbionts on hosts for pest control is a growing research area, but requires knowledge of endosymbionts associated with the target pest population. In this study, we used molecular methods to screen southern Mexico populations of two species of tephritid fruit fly pests, Anastrepha ludens and A. striata , for heritable bacteria. The only heritable endosymbiont found was Wolbachia in A. striata . Based on multilocus sequence typing and phylogenetic analyses, this Wolbachia strain is new and belongs to the Wolbachia supergroup B. Wolbachia strains previously reported in members of the genus Anastrepha in South America belong to supergroup A. We discuss the potential implications for pest control of the presence of a different Wolbachia strain in southern Mexico. Content Type Journal Article Pages 1-8 DOI 10.1007/s00284-012-0223-3 Authors Humberto Martínez, Department of Wildlife and Fisheries Sciences, Texas A&M University, College Station, Texas, USA Jorge Toledo, El Colegio de la Frontera Sur, Tapachula, Chiapas, Mexico Pablo Liedo, El Colegio de la Frontera Sur, Tapachula, Chiapas, Mexico Mariana Mateos, Department of Wildlife and Fisheries Sciences, Texas A&M University, College Station, Texas, USA Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Bombyx mori Bidensovirus ( Bm BDV), a bipartite virus possesses two single-stranded linear DNAs (VD1 and VD2) and shows high pathogenic ability to Bombyx mori . Previous research found that the genes of nonstructural protein ns1 and ns2 were in the same transcript. To investigate the mechanism of transcriptional regulation of ns1 and ns2 genes, the 5′-flanking sequence (289 nt) of ns1 gene, encompasses the regions of the common terminal sequence (CTS) and the predicted P5 promoter from the 5′-terminus of the viral genome to the transcription initiation site of the ns1 gene was cloned and fused to the upstream of the luciferase reporter gene. The luciferase reporter assay showed that the 53 nt CTS of VD1 and VD2 can downregulate the activity of P5 by 13.3 %. The comparison in different cell lines showed that P5 possessed high promoter activity in Bm N and Hi5 cell lines. Interestingly, P5 also had high activity in Hela cells, a kind of cancer cell of human. Subsequent truncated promoter analysis showed that the 31 nt (−236 to −206 nt) sequence is very important to P5 for the activity down to 36.5 % after deletion of it. While the activity also remained 26.5 % after the deletion of the TATA box, suggesting that the promoter is TATA independent. Moreover, in order to further understand the activity intensity of P5, a comparison with other three promoters, B. mori actin3 ( Bm -actin3), B. mori nuclear polyhedrosis virus ( Bm NPV) immediate early 1 gene promoter ( Bm NPV-ie-1), and a synthetic promoter (3xP3) was carried out, the result indicated that the activity of P5 was weaker than that of anyone of them. Content Type Journal Article Pages 1-6 DOI 10.1007/s00284-012-0199-z Authors Shoulin Zhu, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Guohui Li, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Zhaoyang Hu, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Keping Chen, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Guangtian Li, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Xuli Guo, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Ying Ma, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Qin Yao, Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang, 212013 Jiangsu, People’s Republic of China Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    In the present study, using the murine monocyte/macrophage cell line RAW264.7 as a model system, we analyzed the phagocytosis rate and the bactericidal capacity of polyunsaturated fatty acids (PUFA)-enriched macrophages against Pseudomonas aeruginosa and Rhodococcus equi . The P. aeruginosa strain ATCC 10145, the virulent R. equi strain ATCC 33701, and the non-virulent R. equi strain ATCC 6939 were examined. Flow cytometric detection of intracellular microorganisms in combination with viability assays were used to determine the impact of PUFA on the number of engulfed, surviving as well as replicating bacteria. Macrophage enrichment with PUFA resulted in an increase of the internalization rate of the microorganisms by the immune cells. Moreover, an impeding action of the unsaturated fatty acids on the intracellular survival rates of the virulent strains P. aeruginosa ATCC 10145 and R. equi ATCC 33701 could be observed. The n-3 fatty acid docosahexaenoic acid (DHA) as well as the n-6 fatty acid arachidonic acid (AA) showed the most pronounced effects. Taken together, our data support the idea of supplementing PUFA to immunocompromised individuals as well as to people suffering from chronic infections with P. aeruginosa or R. equi to improve macrophage phagocytic and microbicidal activity. Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0207-3 Authors Stephanie Adolph, Faculty of Veterinary Medicine, Institute of Physiological Chemistry, University of Leipzig, An den Tierkliniken 1, 04103 Leipzig, Germany Herbert Fuhrmann, Faculty of Veterinary Medicine, Institute of Physiological Chemistry, University of Leipzig, An den Tierkliniken 1, 04103 Leipzig, Germany Julia Schumann, Faculty of Veterinary Medicine, Institute of Physiological Chemistry, University of Leipzig, An den Tierkliniken 1, 04103 Leipzig, Germany Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    Four antagonists bacteria namely, Bacillus megaterium MB3, B. subtilis MB14, B. subtilis MB99 and B. amyloliquefaciens MB101 were able to produce chitinase, β-1,3-glucanase and protease in different range with the presence of Rhizoctonia solani cell wall as a carbon source. Amplification of chitinase ( chiA ) gene of 270 bp and β-1, 3-glucanase gene of 415 bp was given supportive evidence at molecular level of antibiosis. After in vitro screening, all antagonists were tested against R. solani under greenhouse conditions. Root treatment of Bacillus strains showed superior defense during pathogen suppression in terms of chitinase, glucanase, peroxidase, poly phenol oxidase, phenylalanine ammonia-lyase activity and total phenolic content in leaves of tomato. All these enzymes accumulated high in tomato leaves as compared to roots. Pathogenesis-related proteins and defense-related enzymes accumulation was directly correlated with plant protection and greenhouse results indicated that B. amyloliquefaciens MB101- and B. subtilis MB14-treated plants offered 69.76 and 61.51 % disease reductions, respectively, over the infected control. These results established that these organisms have the potential to act as biocontrol agents. This study could be highlighted a mutual importance of liquid formulation of antagonistic Bacillus spp. against root associated sclerotia former pathogen R. solani . Content Type Journal Article Pages 1-7 DOI 10.1007/s00284-012-0160-1 Authors Manoj Kumar Solanki, National Bureau of Agriculturally Important Microorganisms, Kusmaur, Mau, 275101 Uttar Pradesh, India Amrita Shalini Robert, National Bureau of Agriculturally Important Microorganisms, Kusmaur, Mau, 275101 Uttar Pradesh, India Rajesh Kumar Singh, National Bureau of Agriculturally Important Microorganisms, Kusmaur, Mau, 275101 Uttar Pradesh, India Sudheer Kumar, National Bureau of Agriculturally Important Microorganisms, Kusmaur, Mau, 275101 Uttar Pradesh, India Akhilesh Kumar Pandey, Department of Bioscience, Rani Durgavati University, Jabalpur, 482 001 Madhya Pradesh, India Alok K. Srivastava, National Bureau of Agriculturally Important Microorganisms, Kusmaur, Mau, 275101 Uttar Pradesh, India Dilip K. Arora, National Bureau of Agriculturally Important Microorganisms, Kusmaur, Mau, 275101 Uttar Pradesh, India Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651

Description:    The properties of bacterial isolates from polluted environments which are characterized by increased levels of oxidative stress do not reflect only the level of contaminants, but also arise as a consequence of many permanently changed conditions. The survival rate of Comamonas terrigena N3H isolates from an environment with elevated levels of H 2 O 2 is correlated with stimulation of catalase. The response of bacterial catalase to the effect of phenol in exogenous conditions was affected by the presence of an additional contaminant, Cd 2+ . An isolate of Aspergillus niger selected from river sediment containing 363 mg/kg As, 93 mg/kg Sb at pH 5.2–4.8 grew on Czapek-Dox agar ~1.6 times faster than an isolate of the same species from coal dust sediment with approximately the same level of pollution (400 mg/kg As) but somewhat lower pH (3.3–2.8). It also exhibited differences in the microscopic characteristics of its mycelial structures. Both isolates exhibited a higher tolerance to the exogenic toxic effects of metals (As 5+ , Cd 2+ , and Cu 2+ at 5, 25, or 50 mg/L) than a control culture, but the differences in tolerance between them were only slight. These laboratory results suggest that there are complicated relationships which may exist in the “in situ” environment. Content Type Journal Article Pages 1-5 DOI 10.1007/s00284-012-0163-y Authors Bystrík Polek, Institute of Molecular Biology, Slovak Academy of Sciences, 845 51 Bratislava, Slovakia Jana Godočíková, Institute of Molecular Biology, Slovak Academy of Sciences, 845 51 Bratislava, Slovakia Journal Current Microbiology Online ISSN 1432-0991 Print ISSN 0343-8651