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