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Showing papers in "Current Microbiology in 2007"


Journal ArticleDOI
TL;DR: Investigating in vitro the protective effect of commercial probiotic strains alone and in combination on the adhesion of pathogenic strains as Salmonella, Clostridium, and Escherichia coli to pig intestinal mucus obtained from different intestinal regions revealed a better ability to inhibitAdhesion of all pathogens tested to pig intestine mucus than probiotics strains.
Abstract: The aim of this study was to investigate in vitro the protective effect of commercial probiotic strains (Bifidobacterium lactis Bb12 and Lactobacillus rhamnosus LGG) alone and in combination on the adhesion of pathogenic strains as Salmonella, Clostridium, and Escherichia coli to pig intestinal mucus obtained from different intestinal regions. In combination, probiotic strains enhanced each other's adhesion, mainly in large intestinal mucus. Treatment of intestinal mucus with Bb12 and LGG, alone or in combination, significantly reduced (P < 0.05) the adhesion of the tested pathogens. The ability to inhibit pathogen adhesion appears to depend on the specific probiotics and pathogens and on the mucosal site. B. lactis Bb12 and L. rhamnosus LGG in combination revealed a better ability to inhibit adhesion of all pathogens tested to pig intestinal mucus than probiotic strains. Probiotic combinations could be useful for counteracting disease-associated aberrations in intestinal microbiota. Specific protective probiotics could be selected for particular pig pathogens. Probiotic strains from human origin and intended for human use also adhere to pig intestinal mucus and are able to displace and inhibit pathogens.

178 citations


Journal ArticleDOI
TL;DR: The bacterial strain Bacillus subtilis UTM 126 produced antimicrobial activity against pathogenic Vibrio species, including V. alginolyticus, V. parahaemolyticUS, and V. harveyi in shrimp aquaculture.
Abstract: The bacterial strain Bacillus subtilis UTM 126 produced antimicrobial activity against pathogenic Vibrio species, including V. alginolyticus, V. parahaemolyticus, and V. harveyi. The probiotic effect of B. subtilis was tested by feeding juvenile shrimp (Litopenaeus vannamei) food supplemented with B. subtilis (10(5 )CFU/g) for 28 days before an immersion challenge with V. harveyi at 10(5 )CFU/mL for 24 h. The treatment with B. subtilis UTM 126 decreased final mortality to 18.25%, compared with 51.75% in the control group. Bacillus subtilis UTM 126 has potential applications for controlling pathogenic V. harveyi in shrimp aquaculture.

172 citations


Journal ArticleDOI
TL;DR: Striking discrepancies in supergroup designation between MLST and wsp inferences are reported, and a revision of current methods for Wolbachia supergroup typing is proposed.
Abstract: The obligate intracellular bacteria Wolbachia are taxonomically subdivided into eight supergroups (named A-H). Supergroup typing of strains has been mostly based on phylogenetic inference of the Wolbachia surface protein (wsp), a gene that recently has been shown to experience high rates of recombination. This brings into question its suitability not only for microtaxonomy, but also for supergroup classification of the genus. A Multilocus Sequence Typing (MLST) scheme for Wolbachia has recently been developed that types strains based on five conserved genes, thus providing a rigorous supergroup annotation of strains. Here we report striking discrepancies in supergroup designation between MLST and wsp inferences, and propose a revision of current methods for Wolbachia supergroup typing. Transfer of whole wsp gene sequences between supergroups A and B has occurred. Furthermore, as a result of intragenic recombination, wsp phylogeny creates spurious basal lineages that are not supported by MLST. For example, the proposed supergroup G, based upon wsp alone, likely represents only a wsp recombinant clade. Removal of supergroup G is advised until and unless the existence of this lineage is substantiated by other sequence information (e.g., MLST). We recommend a full characterization MLST for a correct strain typing, while, based on the current data set, use of a single MLST gene can be effective for supergroup designation of A and B strains. Finally, we note that the sharing of wsp sequences between A and B strains indicates a strong genetic cohesiveness of Wolbachia strains, supporting designation of these bacteria within the same species, W. pipientis.

170 citations


Journal ArticleDOI
TL;DR: Emulsification properties of the biosurfactant produced were compared to those of commercial emulsifiers and other microbial surfactants and found that ethyl acetate was able to extract crude surfactant material with high product recovery.
Abstract: Candida lipolytica synthesized a surfactant in a cultivation medium supplemented with canola oil and glucose as carbon sources. Measurements of biosurfactant production and surface tension indicated that the biosurfactant was produced at 48 h of fermentation. The surface-active species is constituted by the protein–lipid–polysaccharide complex in nature. The cell-free broth was particularly influenced by the addition of salt, the pH and temperature depending on the emulsified substrate (hexadecane or a vegetable oil). After comparison between ethyl acetate and mixtures of chloroform and methanol as solvent systems for surfactant recovery, it was found that ethyl acetate was able to extract crude surfactant material with high product recovery (8.0 g/L). The isolated biosurfactant decreased the surface tension to values of 30 mN/m at the critical micelle concentration. Emulsification properties of the biosurfactant produced were compared to those of commercial emulsifiers and other microbial surfactants.

146 citations


Journal ArticleDOI
TL;DR: The present observations demonstrated that the chromium-reducing, metal-solubilizing, and plant growth–promoting potentials of the Bacillus strains PSB1, PSB 7, and PSB10 were not adversely affected by thechromium application and, hence, may be applied for raising the productivity of crops under metal-contaminated soils.
Abstract: The plant growth-promoting potentials, production of siderophore and solubilization of insoluble phosphorus (P) and zinc and lead by the chromium (vi) -reducing Bacillus species, PSB 1, PSB 7, and PSB 10, was assessed both in the presence and absence of chromium under in vitro conditions. The Bacillus strains tolerated chromium up to the concentration of 500 (PSB1), 400 (PSB7), and 550 microg ml(-1) (PSB10), respectively, on nutrient agar plates. Bacillus sp. PSB 10 reduced Cr (vi) by 87% at pH 7, which was followed by Bacillus sp. PSB 1 (83%) and PSB 7 (74%) in nutrient broth after 120 h of incubation. A concentration of 50 microg ml(-1) of Cr (vi) was completely reduced by Bacillus sp. PSB 1 and PSB 10 (after 100 h) and PSB 7 (after 120 h). The Bacillus strains PSB 1, PSB 7, and PSB 10 produced 19.3, 17.7, and 17.4 microg ml(-1) of indole acetic acid, respectively, in luria bertani broth at 100 microg ml(-1) of tryptophan, which consistently decreased with an increase in chromium concentration. The Bacillus strains were positive for siderophore, HCN, and ammonia both in the absence and presence of chromium. The Bacillus strains solubilized 375 (PSB 1), 340 (PSB 7), and 379 (PSB 10) microg ml(-1) P, respectively, in Pikovskaya broth devoid of chromium. In contrast, chromium at 150 microg ml(-1) reduced the amount of P solubilized by 17 (PSB 1), 15 (PSB 7), and 9% (PSB 10) compared to control. The tested bacterial strains solubilized a considerable amount of zinc and lead in nutrient broth both in the absence and presence of chromium. Generally, the chromium reduction and the plant growth-promoting potentials of chromium-reducing Bacillus were strongly correlated at the tested concentration of chromium. The present observations demonstrated that the chromium-reducing, metal-solubilizing, and plant growth-promoting potentials of the Bacillus strains PSB1, PSB 7, and PSB10 were not adversely affected by the chromium application and, hence, may be applied for raising the productivity of crops under metal-contaminated soils.

121 citations


Journal ArticleDOI
TL;DR: Investigation of live and dead Spirulina sp.
Abstract: Cadmium is an important environmental pollutant and a potent toxicant to bacteria, algae, and fungi. Mechanisms of Cd+2 toxicity and resistance are variable, depending on the organism. The present work reports the use of live and dead Spirulina sp. for sorption of Cd+2. This investigation shows that this biomass takes up substantial amount of Cd+2 ions. IR spectroscopic study, kinetics models, Langmuir & Freundlich adsorption isotherms, scanning electron microscopic analysis of Spirulina sp., and the Spirulina sp. treated with different metal ions have been employed to understand the sorption mechanism. Infrared spectra of live Spirulina treated with Cd+2 ions for different lengths of time have been taken to understand the time dependency of metal interaction.

109 citations


Journal ArticleDOI
TL;DR: This present study is the first report of a novel bacteriocin, pumilicin 4, produced by B. pumilus that has potential for use as an alternative antibacterial agent for the treatment of infection with MRSA and VRE.
Abstract: A total of 34 bacterial strains with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity were isolated from 69 soil and water samples collected from four areas of Thailand. One strain, WAPB4 identified as Bacillus pumilus, showed remarkable antibacterial activity against MRSA, vancomycin-resistant Enterococcus faecalis (VRE), and several Gram-positive test bacteria. Bacteriocin produced by WAPB4 was designated as pumilicin 4. It was heat stable up to 121 degrees C, 15 min and active within the pH range of 3-9. Its activity disappeared when treated with pronase E, chymotrypsin, and trypsin, demonstrating its proteinaceous nature. At high dosage (80 AU mL(-1)), the effect of pumilicin 4 was bactericidal to both MRSA and VRE. Bacteriostasis was observed for a low dose of bacteriocin (20 AU mL(-1)). Purification of pumilicin 4 was performed by a three-step procedure, i.e., solvent extraction, solid phase extraction, and reversed-phase chromatography. The molecular mass of purified pumilicin 4 as determined by mass spectrometry was 1994.62 Dalton. This present study is the first report of a novel bacteriocin, pumilicin 4, produced by B. pumilus that has potential for use as an alternative antibacterial agent for the treatment of infection with MRSA and VRE.

107 citations


Journal ArticleDOI
TL;DR: Eight native strains tolerant to 1 M NaCl and displaying plant growth promotion and/or biocontrol features were selected for further characterization and showed antagonistic activity against phytopathogenic fungi belonging to Sclerotinia and Fusarium genus.
Abstract: A bacterial collection of approximately one thousand native strains, isolated from saline soils of Cordoba province (Argentina), was established. From this collection, a screening to identify those strains showing plant growth promotion and biocontrol activities, as well as salt tolerance, was performed. Eight native strains tolerant to 1 M NaCl and displaying plant growth promotion and/or biocontrol features were selected for further characterization. Strains MEP2 18, MRP2 26, MEP2 11a, MEP3 1, and MEP3 3b significantly increased the growth of maize seedlings under normal and saline conditions, whereas isolates ARP2 3, AEP1 5, and ARP2 6 were able to increase the root dry weight of agropyre under saline conditions. On the other hand, strains MEP2 18 and ARP2 3 showed antagonistic activity against phytopathogenic fungi belonging to Sclerotinia and Fusarium genus. Antifungal activity was found in cell-free supernatants, and it was heat and protease resistant. Strains MEP218 and ARP23 were identified as Bacillus sp. and strains MEP211a and MEP33b as Ochrobactrum sp. according to the sequence analysis of 16S rRNA gene.

104 citations


Journal ArticleDOI
TL;DR: Two main antifungal substances produced by this strain proved to be phenazine-1-carboxylic acid and 2-hydroxyphenazine with further purification and structure elucidation based on ultraviolet-absorbent spectrum scanning, atmospheric pressure chemical ionization–mass spectrometry (APCI-MS) spectrum, and 1H,13C nuclear magnetic resonance spectrums.
Abstract: A new Pseudomonas strain, designated GP72, was isolated from green pepper rhizosphere and identified as a member of species Pseudomonas chlororaphis based on morphology; conventional biochemical and physiologic tests; Biolog GN system (Biolog Inc., Hayward, CA); and 16S rDNA sequence analysis. The secondary metabolites produced by this strain have shown broad-spectrum antifungal activity against various phytopathogens of agricultural importance in vitro. Two main antifungal substances produced by this strain proved to be phenazine-1-carboxylic acid and 2-hydroxyphenazine with further purification and structure elucidation based on ultraviolet-absorbent spectrum scanning, atmospheric pressure chemical ionization-mass spectrometry (APCI-MS) spectrum, and (1)H,(13)C nuclear magnetic resonance spectrums. Strain GP72 could produce quorum-sensing signaling molecules of N-butanoyl-L-homoserine lactone and N-hexanoyl-L-homoserine lactone, which were found to accumulate with different quantities in King's medium B and pigment producing medium, respectively.

101 citations


Journal ArticleDOI
TL;DR: Three different species of hydrogen cyanide-producing rhizobacteria were tested for their potential to kill O. obesus and were found to be effective in killing the termites under in vitro conditions.
Abstract: The subterranean termite Odontotermes obesus is an important pest of the Indian subcontinent, causing extensive damage to major agricultural crops and forest plantation trees. Control of termites by strategies employing their parasites has limitations because they have evolved a complex social structure, immune responses, and adaptive behavior toward pathogen-infected individuals. Nonparasitic rhizobacteria that produce harmful metabolites might facilitate the biocontrol of termites. In the present investigation, three different species of hydrogen cyanide-producing rhizobacteria were tested for their potential to kill O. obesus. The three bacterial species were found to be effective in killing the termites under in vitro conditions.

94 citations


Journal ArticleDOI
TL;DR: The strain’s phenotypic and biochemical characteristics are consistent with its placement in the Rhizobium genus, and it is confirmed that R. radiobacter and R. rubi are its closest relatives as indicated by 16S rRNA gene–sequence alignments.
Abstract: A Gram-negative, nonpigmented bacterium designated strain B1 was isolated from a laboratory bioreactor that reduced selenate to elemental red selenium (Se0). 16S rRNA gene–sequence alignment identified the isolate as a Rhizobium sp. belonging to the Rhizobium clade, which includes R. daejeonense, R. giardinii, R. undicola, R. larrymoorei, R. radiobacter, R. rubi, and R. vitis.R. radiobacter and R. rubi are its closest relatives as indicated by 16S rRNA gene–sequence alignments, which differ from strain B1 by 2.6% and 2.8%, respectively. Within this group, strains that show variances > 0.8% to 2.2% have been classified as different species. The major cellular fatty acids present in the B1 strain were C16:0 (1.8%), C18:0 (3.38%), 18:0 3-OH (1.6%), 18:1 ω7c (86.8%), 19:0 cycloω8c (1.5%), and summed features 2 (3.8%) and 3 (1.2%). The large amount of 18:1 ω7c present is constant with members of this group of bacteria, but the small amounts of 16:0, 19:0 cycloω8c, and summed feature 3 shows variance from R. radiobacter and R. rubi. The strain’s phenotypic and biochemical characteristics are consistent with its placement in this genus.

Journal ArticleDOI
TL;DR: The construction of six novel plasmid-based IPTG-inducible expression vectors for Bacillus subtilis and related species are described and the versatility of all six vectors was demonstrated by the insertion of several reporter genes and by their regulated overexpression.
Abstract: We describe the construction of six novel plasmid-based IPTG-inducible expression vectors for Bacillus subtilis and related species. While one vector allows intracellular production of recombinant proteins, the second provides a strong secretion signal. The third vector allows addition of the c-Myc epitope tag, and the remaining three vectors provide the purification tags His and Strep. The versatility of all six vectors was demonstrated by the insertion of several reporter genes and by their regulated overexpression. Recombinant proteins with a His- or Strep-tag could be purified to near homogeneity in a single step.

Journal ArticleDOI
TL;DR: Biological pretreatment with the combined strain suspension after the liquid culture improved enzymatic hydrolysis of office paper from municipal wastes and crystallinity in X-ray diffraction patterns for the crystallinity of cellulose in office paper changed after biological pretreatment.
Abstract: The cellulose-hydrolyzing strains, Sphingomonas paucimobilis MK1 and Bacillus circulans MK2, were separated from soil and were grown together in a single culture plate. Growth B. circulans MK2 in liquid culture required symbiosis with S. paucimobilis MK1. Biological pretreatment with the combined strain suspension after the liquid culture improved enzymatic hydrolysis of office paper from municipal wastes. Sugar recovery by S. paucimobilis MK1 (51%) was 1.4 times higher than that of the untreated sample (30%) and in the strain combination with B. circulans MK2, recovery was further improved by 2.5 times (75%). The sugar recovery in maximum condition was enhanced up to 94% for office paper. Furthermore, biological pretreatment effects were confirmed for more than 1 day less time. In X-ray diffraction patterns for the crystallinity of cellulose in office paper changed after biological pretreatment, the crystallinity was increased in comparison to that in untreated paper. The mechanism of biological pretreatment effect was explained by the fact that the strain acted as an endoglucanase, which hydrolyzes amorphous areas randomly.

Journal ArticleDOI
TL;DR: Phylogenetic analyses indicate clustering in the supergroup F and a high genetic relatedness among all scorpion strains as a result of a potential transmission within the host genus, and an older radiation of F-strains with respect to A and B-str strains, followed by limited horizontal transmission across host genera and reduced genetic flux among strains.
Abstract: The presence and distribution of the intracellular bacteria Wolbachia in the arthropod subphylum Chelicerata (including class Arachnida) has not been extensively explored. Here we report the discovery of Wolbachia in scorpions. Five strains found in host species of the genus Opistophthalmus (Southern African burrowing scorpions) have been characterized by Multilocus Sequence Typing and by Wolbachia Surface Protein. Phylogenetic analyses indicate clustering in the supergroup F and a high genetic relatedness among all scorpion strains as a result of a potential transmission within the host genus. The F-group is an uncommon lineage compared to the A and B supergroups, although it is present in a broad range of hosts (including insects, filarial nematodes, and now arachnids) and across a large geographical area (e.g., North America, Africa, Europe, and Australia). It also shows no evidence of recombination and has a significantly higher genetic diversity than supergroup A and B. Overall, this pattern suggests an older radiation of F-strains with respect to A and B-strains, followed by limited horizontal transmission across host genera and reduced genetic flux among strains. A more extensive sampling of supergroup F-strains is required to confirm this scenario.

Journal ArticleDOI
TL;DR: Different carbon (C) sources, mainly carbohydrates and lipids, have been screened for their capacity to support growth and lipase production by Penicillium restrictum in submerged fermentation (SmF) and in solid-state fermentation (SSF).
Abstract: Different carbon (C) sources, mainly carbohydrates and lipids, have been screened for their capacity to support growth and lipase production by Penicillium restrictum in submerged fermentation (SmF) and in solid-state fermentation (SSF). Completely different physiological behaviors were observed after the addition of easily (oleic acid and glucose) and complex (olive oil and starch) assimilable C sources to the liquid and solid media. Maximal lipolytic activities (12.1 U/mL and 17.4 U/g) by P. restrictum were obtained with olive oil in SmF and in SSF, respectively. Biomass levels in SmF (12.2-14.1 mg/mL) and SSF (7.0-8.0 mg/g) did not varied greatly with the distinct C sources used. High lipase production (12.3 U/g) using glucose was only attained in SSF, perhaps due to the ability of this fermentation process to minimize catabolite repression.

Journal ArticleDOI
TL;DR: The vector pRAG5 is suitable for normalized measurements of promoter activities during the growth of bacterial batch cultures because estimation of the GFP-to-red fluorescent protein fluorescence ratio in strains carrying the plasmid pRAG 5 with the tested promoters upstream of gfpuv avoids the influence of plasmids copy number variations on the promoter activity assay.
Abstract: Novel shuttle promoter-probe vectors replicating in Escherichia coli, Corynebacterium glutamicum, and Rhodococcus erythropolis were constructed on the basis of the C. glutamicum plasmid pCG1. The vectors carry reporter genes coding for fluorescent proteins, which allow the measurement of promoter activities in vivo. The promoter-probe vector pPRE11 contains the rsgfp reporter gene, coding for a variant of green fluorescent protein (GFP) with a red-shifted excitation maximum. To ensure efficient expression of the gfp gene in R. erythropolis from the tested promoters, the promoterless gene gfpuv, with 5' end fusion with the initial six codons of the aph gene and upstream insertion of the aph Shine-Dalgarno sequence, was used as a reporter gene in the promoter-probe vector pEPR1. Insertion of the rfp reference gene, coding for a variant of the red fluorescent protein DsRed.T4 and cloned under the strong constitutive C. glutamicum promoter P-45, into the vector pEPR1 resulted in a new-generation promoter-probe vector (pRAG5). All vectors were tested using a set of mutant P-dapA promoters displaying various transcriptional activities. The vector pRAG5 is suitable for normalized measurements of promoter activities during the growth of bacterial batch cultures because estimation of the GFP-to-red fluorescent protein fluorescence ratio in strains carrying the plasmid pRAG5 with the tested promoters upstream of gfpuv avoids the influence of plasmid copy number variations on the promoter activity assay.

Journal ArticleDOI
TL;DR: The cellular response and proteomic analysis of Escherichia coli exposed to tea polyphenols extracted from Korean green tea and results provide clues for understanding the mechanism of TPP-induced stress and cytotoxicity on E. coli.
Abstract: The purpose of this study was to characterize the cellular response and proteomic analysis of Escherichia coli exposed to tea polyphenols (TPP) extracted from Korean green tea (Camellia sinensis L). TPP showed a dose-dependent bactericidal effect on E. coli. Analysis of cell-membrane fatty acids of E. coli cultures treated with TPP identified unique changes in saturated and unsaturated fatty acids, whereas scanning electron microscopic analysis demonstrated the presence of perforations and irregular rod forms with wrinkled surfaces in cells treated with TPP. Two-dimensional polyacrylamide gel electrophoresis of soluble protein fractions from E. coli cultures exposed to TPP showed 17 protein spots increased or decreased by TPP. Nine upregulated proteins were identified (including GroEL and proteins involved in cellular defense, such as GyrA, RpoS, SodC, and EmrK), whereas the expression of eight proteins was downregulated by exposure to TPP (including proteins involved in carbon and energy metabolism, such as Eno, SdhA, and UgpQ, as well as those involved in amino-acid biosynthesis, such as GltK and TyrB). These results provide clues for understanding the mechanism of TPP-induced stress and cytotoxicity on E. coli.

Journal ArticleDOI
TL;DR: According to the results, the Antarctic biosurfactant-producing strain Pantoea sp.
Abstract: The facultative anaerobe Pantoea sp. strain A-13, isolated from ornithogenic soil of Dewart Island (Frazier Islands), Antarctica, produced glycolipid biosurfactants when grown on n-paraffins or kerosene as the sole source of carbon and energy. Hemolysis of erythrocytes, growth inhibition of Bacillus subtilis, and thin-layer chromatography studies have suggested that the secreted glycolipids are rhamnolipids. Glycolipids produced by kerosene-grown cells decreased the surface tension at the air–water interface to 30 mN/m and possessed a low critical micelle concentration value of 40 mg/l, which indicated high surface activity. They efficiently emulsified aromatic hydrocarbons, kerosene, and n-paraffins. Biosurfactant production contributed to an increase in cell hydrophobicity, which correlated with increased growth of the strain on tested hydrocarbons. According to the results, the Antarctic biosurfactant-producing strain Pantoea sp. A-13 appears to be valuable source for application in accelerated environmental bioremediation.

Journal ArticleDOI
TL;DR: It is shown that strains producing multiple β-lactamases are also present in community-acquired bacterial isolates, indicating a common source of acquisition.
Abstract: The occurrence of extended-spectrum-β-lactamase (ESBL)-producing strains in the community was investigated in a private laboratory located in Juiz de Fora, Brazil. All enterobacterial isolates analysed were collected from urine of human patients between the years 2000 and 2002. ESBL production was confirmed by double disk screening, combination disk method, and Etest ESBL strip. The isoelectric point of each β-lactamase was determined in the crude extracts from each isolate. Detection of ESBL genes was performed by polymerase chain reaction and the genetic relatedness of the isolates determined by pulsed-field gel electrophoresis (PFGE). Of the 1,481 isolates, 22 (12 Klebsiella pneumoniae, 7 Escherichia coli, 1 Providencia stuartii, 1 Citrobacter freundii, and 1 Serratia marcescens) were identified as ESBL producers. The frequency of ESBL producers in the community was 1.48%. TEM-type enzymes were identified in 95.4% of the isolates, followed by the SHV type. Seven strains produced CTX-M–type enzymes. This study showed that strains producing multiple β-lactamases are also present in community-acquired bacterial isolates. Multiple strains exhibiting identical PFGE genotypes were found in individual patients indicating a common source of acquisition.

Journal ArticleDOI
TL;DR: It was concluded that the use of more than one screening method is necessary to detect all MRSA isolates in clinical settings, and cefoxitin disk diffusion was found to be the most specific.
Abstract: In the present study, several conventional methods to detect methicillin-resistant Staphylococcus aureus (MRSA) were compared with polymerase chain reaction (PCR) detection of mecA gene–positive isolates. Cefoxitin E-test was also evaluated as a possible phenotypic method of MRSA detection. Oxacillin agar screen and PBP2′ latex agglutination methods were found to be more sensitive than oxacillin and cefoxitin disk-diffusion methods. Cefoxitin disk diffusion was found to be the most specific. A combination of oxacillin agar screening with cefoxitin disk diffusion, or oxacillin disk diffusion with PBP2′, improved sensitivity and specificity. Cefoxitin E-test with the current break points had low sensitivity and specificity (33.3% and 75%, respectively) for the detection of MRSA. However, changing the break points to ≤ 4 μg/ml and to ≥ 6 μg/ml for sensitive and resistant, respectively, greatly improved both. Changing the 30-μg cefoxitin disk-diffusion break points to ≤ 21 mm for resistant slightly improved sensitivity but had no effect on specificity. It was therefore concluded that the use of more than one screening method is necessary to detect all MRSA isolates in clinical settings.

Journal ArticleDOI
TL;DR: The photosynthetic bacteria Rhodobacter capsulatus was screened and found to have strong ability to adsorb Au(III), which might be the mechanism of photosynthtic bacteria metal tolerance.
Abstract: Biosorption has been shown to be an eco-friendly approach to remove heavy metal ions. In this study, the photosynthetic bacteria Rhodobacter capsulatus was screened and found to have strong ability to adsorb Au(III). The maximum specific uptake of living cells was over 92.43 mg HAuCl4/g dry weight of cell in the logarithmic phase. Biosorpion ability would be enhanced by an acidic environment. As the main cations, during biosorption the quantity of Mg2+ exchanged was more than Na+. Biosorbed Au(III) could be reduced by carotenoid and enzymes embedded and/or excreted by R. capsulatus, which might be the mechanism of photosynthtic bacteria metal tolerance.

Journal ArticleDOI
TL;DR: The results highlighting the relevance of unexplored microbes from cold environments of Western Himalayas for the isolation of protease enzymes active at wide range of temperature and pH suggests that the protease isolated from psychrotrophic Exiguobacterium sp.
Abstract: Out of nine psychrotrophic bacterial strains isolated from cold environments of the Western Himalayas, SKPB5 was selected for protease purification and characterization because it had the largest zone of clearance on plate assay. On the basis of the phenotypic and biochemical characterization and 16S rRNA gene-sequencing studies, isolate was identified as Exiguobacterium sp. SKPB5. The protease was purified near to homogeneity with a purification fold of 7.1, and its molecular weight was determined to be 36 kDa. The enzyme exhibited maximum stability at 50 degrees C and an optimal pH of 8.0. Metal ions Mg2+, Ca2+, Zn2+, and Mn2+ enhanced the enzyme activity, whereas Cu2+ had no effect. Phenylmethanesulfonyl fluoride and ethylenediaminetetraacetic acid did not show any effect on the activity of the enzyme, whereas a 20% increase in activity was observed when it was incubated in presence of reducing agents such as beta-mercaptoethanol and dithiothreitol. This suggests that the protease isolated from psychrotrophic Exiguobacterium sp. SKPB5 belongs to the cysteine family. The results highlight the relevance of unexplored microbes from cold environments of Western Himalayas for the isolation of protease enzymes active at wide range of temperature and pH.

Journal ArticleDOI
TL;DR: Biosurfactants produced from cost-free by-products combines waste minimization with economic potential bioremediation process and did not have toxic effects over the microbial population.
Abstract: A crude biosurfactant solution was produced by Pseudomonas aeruginosa growing on agroindustrial wastes as the substrate and used to study its effect on hydrocarbon biodegradation by the indigenous soil microflora under laboratory conditions. Two concentrations were studied at first and 1 mg of biosurfactant/g of soil showed to be the most efficient for the total petroleum hydrocarbon reduction, which reached 85% at the first 20 days in soil microcosms. Respirometric and microbial analyses showed that the biosurfactant added did not have toxic effects over the microbial population. The use of a biosurfactant for bioremediation has been limited because of its high cost production. Biosurfactants produced from cost-free by-products combines waste minimization with economic potential bioremediation process.

Journal ArticleDOI
TL;DR: Four defective (AFM−) mutants of Paenibacillus sp.
Abstract: Four defective (AFM−) mutants of Paenibacillus sp. HKA-15 that no longer produced the peptide antifungal metabolites were developed through ethyl methane sulfonate (EMS) mutagenesis and used for in vivo experimentation. Reduced percentage of seed germination by mutants DM1 and DM2 (22.5% and 25%, respectively) and a high percent of disease incidence (69.3% and 67%, respectively) compared to wild-strain HKA-15 (80% seed germination and 27% disease incidence) indirectly indicated the role of peptide metabolite on disease suppression. Plants treated with AFM− clones showed stunted growth and the presence of pepperlike microsclerotia in the stem tissues. Light and scanning electron microscopic studies clearly showed the effect of peptide antibiosis on hyphal morphology. Exposure to crude extracts of antibiotics produced abnormal contraction of fungal cytoplasm, granulation, and fragmentation of hyphal mycelia and cell lysis. The presence of bacterial cells in the lumen of degrading fungal mycelium suggested a direct involvement of Paenibacillus sp. HKA-15 in the lysis of Rhizoctonia bataticola.

Journal ArticleDOI
TL;DR: Bacterial isolates screened for their mineral phosphate–solubilizing (MPS) ability on Pikovskaya and National Botanical Research Institute’s phosphate (NBRIP) agar emerged as the best solubilizer and would be a useful microbial inoculant in groundnut cultivation.
Abstract: Twenty-three bacterial isolates were screened for their mineral phosphate–solubilizing (MPS) ability on Pikovskaya and National Botanical Research Institute’s phosphate (NBRIP) agar. The majority of the isolates exhibited a strong ability to solubilize hydroxyapatite in both solid and liquid media. The solubilization in liquid medium corresponded with a decrease in the pH of the medium. Serratia marcescens GPS-5, known for its biocontrol of late leaf spot in groundnut, emerged as the best solubilizer. S. marcescens GPS-5 was subjected to ethyl methanesulfonate (EMS) mutagenesis, and a total of 1700 mutants, resulting after 45 minutes of exposure, were screened on buffered NBRIP medium for alterations in MPS ability compared with that of the wild type. Seven mutants with increased (increased-MPS mutants) and 6 mutants with decreased (decreased-MPS mutants) MPS ability were isolated. All seven increased-MPS mutants were efficient at solubilizing phosphate in both solid and liquid NBRIP medium. Among the increased-MPS mutants, EMS XVIII Sm-35 showed the maximum (40%) increase in the amount of phosphate released in liquid medium compared with wild-type S. marcescens GPS-5, therefore, it would be a useful microbial inoculant in groundnut cultivation. EMS III Sm W, a nonpigmented mutant, showed the lowest solubilization of phosphate among the 6 decreased-MPS mutants.

Journal ArticleDOI
TL;DR: The purified killer toxin actively hydrolyzed laminarin and killed the whole cells of the pathogenic yeast in crab and was strongly inhibited by phenylmethanesulphonyl fluoride, iodoacetic acid, ethylenediaminetetraacetic Acid, and 1,10-phenanthroline.
Abstract: The molecular mass of the purified killer toxin from the marine killer yeast YF07b was estimated to be 47.0 kDa. The optimal pH and temperature of the purified killer toxin were 4.5 and 40 degrees C, respectively. The toxin was activated by Ca(2+), K(+), Na(+), Mg(2+), Na(+), and Co(2+). However, Fe(2+), Fe(3+), Hg(2+), Cu(2+), Mn(2+), Zn(2+), and Ag(+) acted as inhibitors in decreasing activity of the toxin. The toxin was strongly inhibited by phenylmethanesulphonyl fluoride (PMSF), iodoacetic acid, ethylenediaminetetraacetic acid, and 1,10-phenanthroline. The Km of the toxin for laminarin was 1.17 g L(-1). The toxin also actively hydrolyzed laminarin and killed the whole cells of the pathogenic yeast in crab.

Journal ArticleDOI
TL;DR: The isolate isolated by conventional enrichment process from soils of Ocimum field showed potential to be a good candidate for biotechnological production of vanillin from isoeugenol and further studies for standardization and optimization for higher yield needs to be investigated.
Abstract: Vanillin is undoubtedly one of the most popular and widely used flavoring agents in the world. Taking into consideration the worldwide demand for natural vanillin and its limited supply, alternative routes for its production including biotransformation are being constantly explored. In this regard, a novel soil bacterium capable of converting isoeugenol to vanillin was isolated by conventional enrichment process from soils of Ocimum field. On the basis of morphological and physiochemical characteristics and 16S rRNA gene sequence analysis, the isolate was identified as Pseudomonas chlororaphis CDAE5 (EMBL # AM158279). Vanillin formation was analyzed by gas chromatography (GC), and its structure was confirmed by GC-mass spectrometry and nuclear magnetic resonance. After 24-h reaction, the vanillin concentration reached 1.2 g L(-1) from 10 g L(-1) isoeugenol in 20-mL reaction solution at 25 degrees C and 180 rpm. The strain showed potential to be a good candidate for biotechnological production of vanillin from isoeugenol. Further studies for standardization and optimization for higher yield of vanillin production needs to be investigated.

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TL;DR: It was concluded that M. aeruginosa requires higher iron concentration for physiological and biochemical processes compared with M. wesenbergii, but its tolerance against too high a concentration of iron is weaker than M. Wesenberg ii.
Abstract: Changes in growth, photosynthetic pigments, and photosystem II (PS II) photochemical efficiency as well as production of siderophores of Microcystis aeruginosa and Microcystis wesenbergii were determined in this experiment. Results showed growths of M. aeruginosa and M. wesenbergii, measured by means of optical density at 665 nm, were severely inhibited under an iron-limited condition, whereas they thrived under an iron-replete condition. The contents of chlorophyll-a, carotenoid, phycocyanin, and allophycocyanin under an iron-limited condition were lower than those under an iron-replete condition, and they all reached maximal contents on day 4 under the iron-limited condition. PS II photochemical efficiencies (maximal PS II quantum yield), saturating light levels (I-k ) and maximal electron transport rates (ETRmax) of M. aeruginosa and M. wesenbergii declined sharply under the iron-limited condition. The PS II photochemical efficiency and ETRmax of M. aeruginosa rose , whereas in the strain of M. wesenbergii, they declined gradually under the iron-replete condition. In addition, I-k of M. aeruginosa and M. wesenbergii under the iron-replete condition did not change obviously. Siderophore production of M. aeruginosa was higher than that of M. wesenbergii under the iron-limited condition. It was concluded that M. aeruginosa requires higher iron concentration for physiological and biochemical processes compared with M. wesenbergii, but its tolerance against too high a concentration of iron is weaker than M. wesenbergii.

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TL;DR: 16S rRNA gene-based analysis of the isolates showed that the bacteria with similar-resistance physiologies clustered together and belonged to related genera, indicating a divergent evolution of the salt tolerance and radiation resistance.
Abstract: Isolation of five ionizing radiation (IR)-resistant bacteria by screening of isolates from various habitats classified as common and stressed is reported. IR-resistant isolates exhibited varying degrees of resistance to γ-radiation and were classified as highly and moderately radiation resistant. Resistance to ultraviolet (UV) radiation correlated well with γ-radiation resistance, whereas a comparable desiccation resistance for all the highly and moderately radiation-resistant isolates was observed. However, salt tolerance failed to correlate with IR resistance, indicating a divergent evolution of the salt tolerance and radiation resistance. Characterization of isolates by the amplified rDNA restriction analysis profiling attested to the clustering of these isolates with their stress phenotype. 16S rRNA gene-based analysis of the isolates showed that the bacteria with similar-resistance physiologies clustered together and belonged to related genera. Hydrogen peroxide resistance and mitomycin survival patterns of the isolates indicated the roles of oxidative-stress tolerance in desiccation survival and recombination repair in higher radiation resistance, respectively.

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TL;DR: Analysis of peeling black crusts from modern and historic buildings in Campeche, Mexico, from a gravestone on the island of Dom Khon, Lao, and from the Anglican cathedral in Belize City demonstrate that, unlike chemically formed thickBlack crusts found in polluted atmospheres, thinblack crusts in clean environments may be predominantly composed of filamentous cyanobacteria.
Abstract: Samples of peeling black crusts from modern and historic buildings in Campeche, Mexico, from a gravestone on the island of Dom Khon, Lao, and from the Anglican cathedral in Belize City were analyzed microbiologically, by scanning electron microscopy plus electron dispersive spectroscopy (EDS) and for pigment composition. In all cases, the surface was covered by a thick mat of cyanobacteria with dark brown sheaths. These were filamentous organisms of the genera Scytonema or Fischerella/Mastigocladus, except for one sample, where coccoid cyanobacteria of Subsection II were predominant. Fungi were not present at all sites and, where seen, were not the major biomass. High scytonemin:chlorophyll a ratios correlated with the dark pigmentation of the cyanobacterial cells and indicated the stressful conditions under which these organisms were living (high temperatures and ultraviolet levels, frequent desiccation). The absence, or low levels, of sulfur in the biofilms confirmed that there was little urban pollution at the sites and the EDS analysis showed that the black coloration was caused solely by cell pigmentation; no dark-colored elements were present at high concentrations. These results demonstrate that, unlike chemically formed thick black crusts found in polluted atmospheres, thin black crusts (which could be called patinas) in clean environments may be predominantly composed of filamentous cyanobacteria.