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Sample records for bacillus megaterium

  1. Systems biology of recombinant protein production using Bacillus megaterium.

    Science.gov (United States)

    Biedendieck, Rebekka; Borgmeier, Claudia; Bunk, Boyke; Stammen, Simon; Scherling, Christian; Meinhardt, Friedhelm; Wittmann, Christoph; Jahn, Dieter

    2011-01-01

    The Gram-negative bacterium Escherichia coli is the most widely used production host for recombinant proteins in both academia and industry. The Gram-positive bacterium Bacillus megaterium represents an increasingly used alternative for high yield intra- and extracellular protein synthesis. During the past two decades, multiple tools including gene expression plasmids and production strains have been developed. Introduction of free replicating and integrative plasmids into B. megaterium is possible via protoplasts transformation or transconjugation. Using His(6)- and StrepII affinity tags, the intra- or extracellular produced proteins can easily be purified in one-step procedures. Different gene expression systems based on the xylose controlled promoter P(xylA) and various phage RNA polymerase (T7, SP6, K1E) driven systems enable B. megaterium to produce up to 1.25g of recombinant protein per liter. Biomass concentrations of up to 80g/l can be achieved by high cell density cultivations in bioreactors. Gene knockouts and gene replacements in B. megaterium are possible via an optimized gene disruption system. For a safe application in industry, sporulation and protease-deficient as well as UV-sensitive mutants are available. With the help of the recently published B. megaterium genome sequence, it is possible to characterize bottle necks in the protein production process via systems biology approaches based on transcriptome, proteome, metabolome, and fluxome data. The bioinformatical platform (Megabac, http://www.megabac.tu-bs.de) integrates obtained theoretical and experimental data. PMID:21943898

  2. BIOACCUMULATION OF HEAVY METALS BY BACILLUS MEGATERIUM FROM PHOSPHOGYPSUM WASTE

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    IOANA ADRIANA STEFANESCU

    2015-05-01

    Full Text Available The aim of present study was to characterize the bioaccumulation capacity of heavy metals by Bacillus megaterium from phosphogypsum waste. The Bacillus megaterium strain (BM30 was isolated from soil near the phosphogypsum (PG dump. For the bioaccumulation quantification produced by BM30 strain were used three experimental treatments respectively with 2, 6 and 10 gL-1 PG. Cellular biomass samples were collected punctually at ages corresponding to the three stages of the development cycle of the microorganism: exponential phase, stationary phase and decline phase and the heavy metals concentrations were measured by atomic absorption spectroscopy. The bioaccumulation yields in cell biomass, relative to the total amount of analyte introduced in the reaction medium were between 20 - 80 %, the lowest value was recorded by Cu and highest by Mn. The study results indicated that the isolated strain near the dump PG, BM30, bioaccumulate heavy metals monitored in cell biomass in the order Cu > Fe > Zn = Mn.

  3. Production of recombinant antibody fragments in Bacillus megaterium

    OpenAIRE

    Jahn Dieter; Schirrmann Thomas; Biedendieck Rebekka; Roth Andreas; Hust Michael; Jordan Eva; Dübel Stefan

    2007-01-01

    Abstract Background Recombinant antibodies are essential reagents for research, diagnostics and therapy. The well established production host Escherichia coli relies on the secretion into the periplasmic space for antibody synthesis. Due to the outer membrane of Gram-negative bacteria, only a fraction of this material reaches the medium. Recently, the Gram-positive bacterium Bacillus megaterium was shown to efficiently secrete recombinant proteins into the growth medium. Here we evaluated B. ...

  4. Carbonate biomineralization induced by soil bacterium Bacillus megaterium

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    Lian, Bin; Hu, Qiaona; Chen, Jun; Ji, Junfeng; Teng, H. Henry

    2006-11-01

    Biogenic carbonates spawned from microbial activities are common occurrences in soils. Here, we investigate the carbonate biomineralization mediated by the bacterium Bacillus megaterium, a dominant strain separated from a loess profile in China. Upon completing bacterial cultivation, the ensuring products are centrifuged, and the resultant supernatant and the concentrated bacterial sludge as well as the un-separated culture are added separately into a Ca-CO 3 containing solution for crystallization experiments. Results of XRD and SEM analysis indicate that calcite is the dominant mineral phase formed when the bacteria are present. When the supernatant alone is used, however, a significant portion of vaterite is also precipitated. Experimental results further reveal that the bacteria have a strong tendency to colonize the center area of the calcite {1 0 1¯ 4} faces. Observed crystal morphology suggests that the bacterial colony may promote the growth normal to each individual {1 0 1¯ 4} face of calcite when the cell concentration is high, but may retard it or even cause dissolution of the immediate substrate surfaces when the concentration is low. SEM images taken at earlier stages of the crystallization experiments demonstrate the nucleation of calcite on the bacterial cell walls but do not show obvious morphological changes on the nanometer- to submicron-sized nuclei. δ 13C measurements unveil that the crystals grown in the presence of bacteria are further enriched in the heavy carbon isotope, implying that the bacterial metabolism may not be the carbon sources for the mineralization. Based upon these findings, we propose a mechanism for the B. megaterium mediated calcite mineralization and conclude that the whole process involves epi- and inter-cellular growth in the local microenvironments whose conditions may be controlled by cell sequestration and proton pumping during bacterial respiration.

  5. Isolation and genetic characterizations of Bacillus megaterium cobalamin biosynthesis-deficient mutants.

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    Wolf, J B; Brey, R N

    1986-01-01

    Ethanolamine is deaminated by the action of ethanolamine ammonia-lyase (EC 4.3.1.7), an adenosylcobalamin-dependent enzyme. Consequently, to grow on ethanolamine as a sole nitrogen source, Bacillus megaterium requires vitamin B12. Identification of B. megaterium mutants deficient for growth on ethanolamine as the sole nitrogen source yielded a total of 34 vitamin B12 auxotrophs. The vitamin B12 auxotrophs were divided into two major phenotypic groups: Cob mutants, which could use cobinamide o...

  6. Optimization of polyphosphate production by Bacillus megaterium strain G11

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    Giti Emtiazi

    2013-01-01

    Full Text Available Introduction: Polyphosphates, also called volutin granules, are linear polymers from orthophosphates linked by energy-rich phosphoanhydride bands that have been seen in bacteria, yeasts, fungi, plants and animals. These polymers are completely safe and nontoxic, and have numerous applications in food and drug industries.Materials and methods: Due to the great importance and wide range of the utilization of these polymers in various industries, several factors such as various carbon sources, carbon source concentration and phosphorus concentration were studied and optimized. In order to increase polyphosphate production in Bacillus megaterium strain G11. The optimization process was carried out with determination of the amount of polyphosphate accumulated in cell and phosphorus removed from the medium. One-way ANOVA and Tukey tests were used in order to determine whether there was a significant difference between data obtained in this research.Results: Growth of B. megaterium in the presence of sucrose (OD=3.026 was better than glucose (OD=2.616 whereas polyphosphate production and phosphorus removal from medium were higher in the presence of glucose (0.033 g g-1 dry cell weight and 1.61 g l-1, respectively. On the other hand, polyphosphate production and phosphorus removal from medium coordinately were decreased with increasing glucose concentration. Furthermore, in studying the effects of phosphorus, we faced two phases of rising and falling. Actually, the increase of phosphorus concentration (0.25-1 g l-1 in medium caused an increase in polyphosphate production and phosphorus removal from medium whereas both of them were decreased with a more increase in amount of phosphorus (1-4 g l-1. One-way ANOVA and Tukey tests showed that there was a significant difference (P<0.01 between data obtained at each optimization step and the best glucose and dipotassium phosphate concentrations for polyphosphate production were 5 and 0.5 g l-1 respectively

  7. Biocontrol of Aspergillus flavus on peanut kernels by use of a strain of marine Bacillus megaterium.

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    Kong, Qing; Shan, Shihua; Liu, Qizheng; Wang, Xiudan; Yu, Fangtang

    2010-04-30

    A strain of marine Bacillus megaterium isolated from the Yellow Sea of East China was evaluated for its activity in reducing postharvest decay of peanut kernels caused by Aspergillus flavus in in vitro and in vivo tests. The results showed that the concentrations of antagonist had a significant effect on biocontrol effectiveness in vivo: when the concentration of the washed bacteria cell suspension was used at 1x10(9)CFU/ml, the percentage rate of rot of peanut kernels was 31.67%+/-2.89%, which was markedly lower than that treated with water (the control) after 7days of incubation at 28 degrees C. The results also showed that unwashed cell culture of B. megaterium was as effective as the washed cell suspension, and better biocontrol was obtained when longer incubation time of B. megaterium was applied. When the incubation time of B. megaterium was 60-h, the rate of decay declined to 41.67%+/-2.89%. Furthermore, relative to the expression of 18S rRNA, the mRNA abundances of aflR gene and aflS gene in the experiment group were 0.28+/-0.03 and 0.024+/-0.005 respectively, indicating that this strain of B. megaterium could significantly reduce the biosynthesis of aflatoxins and expression of aflR gene and aflS gene (pagainst postharvest fungal disease caused by A. flavus. PMID:20156660

  8. Involvement of plant endogenous ABA in Bacillus megaterium PGPR activity in tomato plants

    OpenAIRE

    Porcel, Rosa; Zamarreño, Ángel M.; García-Mina, José M.; AROCA, RICARDO

    2014-01-01

    Abstract Background Plant growth-promoting rhizobacteria (PGPR) are naturally occurring soil bacteria which benefit plants by improving plant productivity and immunity. The mechanisms involved in these processes include the regulation of plant hormone levels such as ethylene and abscisic acid (ABA). The aim of the present study was to determine whether the activity of Bacillus megaterium PGPR is affected by the endogenous ABA content of the host plant. The ABA-deficient tomato mutants flacca ...

  9. Study on the Effect of Mutated Bacillus megaterium in Two-Stage Fermentation of Vitamin C

    Institute of Scientific and Technical Information of China (English)

    吕树娟; 王军; 姚建铭; 余增亮

    2003-01-01

    Bacillus megaterium as a companion strain in two-stage fermentation of vitamin Ccould secrete some active substances to spur growth of Gluconobacter oxydans to produce 2-KLG.In the fermenting system where Gluconobacter oxydans was combined with GB82-a mutatedstrain of B. megaterium by ion implantation, the amount of 2-KLG harvested was larger thanthat produced by the original B. megaterium BP52[1] being substituted for GB82. In this paper,we studied the effect of the active substances secreted by GB82 to enhance the capability ofGluconobacter oxydans to produce 2-KLG. The supernate of GB82 sampled at different cultivationtimes all had much more activity to spur Gluconobacter oxydans to yield 2-KLG than that of theoriginal B. megaterium, which might be due to the genetic changes in the active componentscaused by ion implantation. Furthermore, the active substances of GB82's supernate would losea part of its activity in extreme environments, which is typical of some proteins.

  10. Synthetic Quorum Sensing and Cell-Cell Communication in Gram-Positive Bacillus megaterium.

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    Marchand, Nicholas; Collins, Cynthia H

    2016-07-15

    The components of natural quorum-sensing (QS) systems can be used to engineer synthetic communication systems that regulate gene expression in response to chemical signals. We have used the machinery from the peptide-based agr QS system from Staphylococcus aureus to engineer a synthetic QS system in Bacillus megaterium to enable autoinduction of a target gene at high cell densities. Growth and gene expression from these synthetic QS cells were characterized in both complex and minimal media. We also split the signal production and sensing components between two strains of B. megaterium to produce sender and receiver cells and characterized the resulting communication in liquid media and on semisolid agar. The system described in this work represents the first synthetic QS and cell-cell communication system that has been engineered to function in a Gram-positive host, and it has the potential to enable the generation of dynamic gene regulatory networks in B. megaterium and other Gram-positive organisms. PMID:26203497

  11. A fuzzy logic algorithm for identification of the harvesting threshold during PGA production by Bacillus megaterium

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    E. R. Nucci

    2005-12-01

    Full Text Available Penicillin G acylase (PGA is an important enzyme used as biocatalyst in the production of semisynthetic beta-lactam antibiotics. Many microorganisms produce this enzyme and recombinant Escherichia coli has been preferred use for industrial applications. Bacillus megaterium is one of the microorganisms that excretes this enzyme into the medium. As a consequence, separation and purification steps are simplified. On-line measurement of enzyme activity during cultivation using in-situ sensors is a difficult task in the industrial environment due to the lack of robust and inexpensive instrumentation. This work presents the results of a fuzzy logic algorithm used to determine the moment of maximum enzyme concentration during Bacillus megaterium cultivations in an aerated and stirred, automated lab-scale bioreactor. The fuzzy algorithm was written in Fortran, compiled as a dynamic link library and implemented on a platform developed in MS-Visual Basic. Data were exchanged in real time between the platform and the supervisory system, which was coupled to the bioreactor. It was possible to determine the moment at which maximum enzyme activity was reached in several bioreactor assays. At this point, the end of the process was indicated to the operator. The results illustrate the importance of using reliable computational intelligence-based algorithms in biochemical reactions.

  12. Expression of recombinant Clostridium difficile toxin A and B in Bacillus megaterium

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    Nie Weijia

    2008-11-01

    Full Text Available Abstract Background Major Clostridium difficile virulence factors are the exotoxins TcdA and TcdB. Due to the large size and poor stability of the proteins, the active recombinant TcdA and TcdB have been difficult to produce. Results The toxin genes tcdA and tcdB were amplified by PCR using chromosomal DNA from a toxigenic strain as a template, and cloned into a shuttle vector pHis1522. The sequences of both tcdA and tcdB genes in the vector have been verified by DNA sequencing. The constructs were transformed into B. megaterium protoplasts and the protein expression was controlled under a xylose promoter. The recombinant toxins (rTcdA and rTcdB were purified from bacterial crude extracts. Approximately 5 – 10 mg of highly purified recombinant toxins were obtained from one liter of bacterial culture. The resulting rTcdA and rTcdB had similar molecular masses to the native toxins, and their biological activities were found to be similar to their native counterparts after an extensive examination. Conclusion We have generated the full length and active recombinant TcdA and TcdB in Bacillus megaterium.

  13. Preparation and evaluation of Bacillus megaterium-alginate microcapsules for control of rice sheath blight disease.

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    Wiwattanapatapee, R; Chumthong, A; Pengnoo, A; Kanjanamaneesathian, M

    2013-08-01

    Bacillus megaterium encapsulated in calcium alginate microcapsules was prepared and tested for its efficacy against sheath blight disease of rice. In laboratory conditions, the aqueous suspension (1:100, v/v in potato dextrose agar) of the bacterial microcapsules (10(10) spores/ml) inhibited mycelial growth of Rhizoctonia solani (>99 %) after the microcapsules were produced and stored for 12 months at room temperature (28 ± 2 °C). The survival of the bacterium in the microcapsules in response to ultraviolet (u.v.) irradiation and high temperature was investigated. The survivability of the bacterium in the encapsulated form was greater than that of the fresh cells when it was subjected to u.v. (20-W General electric u.v. lamp from a 25 cm distance for 48 h) and a high temperature treatment (80 °C for 48 h). Cells of the bacterium were detected by scanning electron microscope on both the leaf sheath and the leaf blade (in pot tests in a greenhouse) after spraying encapsulated product. The number of bacteria on the surface of both rice tissues (5 Log. number/g of plant) after spraying with encapsulated product was not significantly different from that after spraying with fresh cells onto the rice seedlings. Spraying the encapsulated B. megaterium on rice plants in the greenhouse was as effective as spraying a chemical fungicide for suppressing rice sheath blight disease. PMID:23508397

  14. Metabolic engineering of cobalamin (vitamin B12) production in Bacillus megaterium.

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    Biedendieck, Rebekka; Malten, Marco; Barg, Heiko; Bunk, Boyke; Martens, Jan-Henning; Deery, Evelyne; Leech, Helen; Warren, Martin J; Jahn, Dieter

    2010-01-01

    Cobalamin (vitamin B(12)) production in Bacillus megaterium has served as a model system for the systematic evaluation of single and multiple directed molecular and genetic optimization strategies. Plasmid and genome-based overexpression of genes involved in vitamin B(12) biosynthesis, including cbiX, sirA, modified hemA, the operons hemAXCDBL and cbiXJCDETLFGAcysG(A)cbiYbtuR, and the regulatory gene fnr, significantly increased cobalamin production. To reduce flux along the heme branch of the tetrapyrrole pathway, an antisense RNA strategy involving silencing of the hemZ gene encoding coproporphyrinogen III oxidase was successfully employed. Feedback inhibition of the initial enzyme of the tetrapyrrole biosynthesis, HemA, by heme was overcome by stabilized enzyme overproduction. Similarly, the removal of the B(12) riboswitch upstream of the cbiXJCDETLFGAcysG(A)cbiYbtuR operon and the recombinant production of three different vitamin B(12) binding proteins (glutamate mutase GlmS, ribonucleotide triphosphate reductase RtpR and methionine synthase MetH) partly abolished B(12)-dependent feedback inhibition. All these strategies increased cobalamin production in B. megaterium. Finally, combinations of these strategies enhanced the overall intracellular vitamin B(12) concentrations but also reduced the volumetric cellular amounts by placing the organism under metabolic stress. PMID:21255303

  15. Produksi Enzim Amilolitik dari Bacillus megaterium Menggunakan Variasi Kadar Pati Sagu (Metroxylon sp.

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    Sandra Madonna

    2016-03-01

    Full Text Available The application of enzymes as biocatalysts for the industries in Indonesia has increased. Among the enzymes that are needed in Indonesia, one of which amylolytic enzymes. Amylolytic enzymes constitute a group of enzymes that catalyze the hydrolysis of starch into simple sugars consisting of glucose units. In this study amylolytic enzyme isolated from the bacterium Bacillus megaterium. Enzyme production was submerged fermentation method for 14 hours using sagostarch Metroxylon sp. varies. Measurement of enzyme activity was determined by the method amylolytik Somogy-Nelson. Research results showed that2 % (w/v of sago starchis the optimum consentration in media with highest amylolytic enzyme activity that is equal to 0.076 units/ml and sugar medium formed by181.254ug/ml in the fermentation medium.

  16. Polyhydroxybutyrate production from oil palm empty fruit bunch using Bacillus megaterium R11.

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    Zhang, Youhong; Sun, Wandong; Wang, Hengwei; Geng, Anli

    2013-11-01

    Oil palm empty fruit bunch (OPEFB), contains abundant cellulose and hemicelluloses and can be used as a renewable resource for fuel and chemical production. This study, as the first attempt, aims to convert OPEFB derived sugars to polyhydroxybutyrate (PHB). OPEFB collected from a Malaysia palm oil refinery plant was chemically pretreated and enzymatically hydrolyzed by an in-house prepared cellulase cocktail. The PHB producer, Bacillus megaterium R11, was isolated in Singapore and could accumulate PHB up to 51.3% of its cell dry weight (CDW) from both glucose and xylose. Tryptone was identified as its best nitrogen source. PHB content and production reached 58.5% and 9.32 g/L, respectively, for an overall OPEFB sugar concentration of 45 g/L. These respectively reached 51.6% and 12.48 g/L for OPEFB hydrolysate containing 60 g/L sugar with a productivity of 0.260 g/L/h. PMID:24001560

  17. Utilization of Sparingly Soluble Phosphate by Red Clover in Association with Glomus mosseae and Bacillus megaterium

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A pot experiment was conducted to investigate the mobilization of sparingly soluble inorganic and organic sources of phosphorus (P) by red clover (Trifolium pratense L.) whose roots were colonized by the arbuscular mycorrhizal (AM) fungus Glomus mosseae and in association with the phosphate-solubilizing (PS) bacterium Bacillus megaterium ACCC10010. Phosphate-solubilizing bacteria and rock phosphate had a synergistic effect on the colonization of plant roots by the AM fungus. There was a positive interaction between the PS bacterium and the AM fungus in mobilization of rock phosphate, leading to improved plant P nutrition. In dual inoculation with the AM fungus and the PS bacterium, the main contribution to plant P nutrition was made by the AM fungus. Application of P to the low P soil increased phosphatase activity in the rhizosphere. Alkaline phosphatase activity was significantly promoted by inoculation with either the PS bacterium or the AM fungus.

  18. Purification and characterization of a Bacillus megaterium disulfide reductase specific for disulfides containing pantethine 4',4"-diphosphate.

    OpenAIRE

    Swerdlow, R D; Setlow, P

    1983-01-01

    An NADH-linked disulfide reductase specific for disulfides containing pantethine 4',4"-diphosphate moieties was purified 23,000-fold to homogeneity from spores of Bacillus megaterium. The enzyme had a native molecular weight of 122,000 with two apparently identical subunits, contained one molecule of flavin adenine dinucleotide per subunit, and was inhibited by the vicinal dithiol reagent arsenite. The enzyme was active only on disulfides containing pantethine 4',4"-diphosphate moieties, incl...

  19. [Interspecific protoplast fusion between Bacillus thuringensis Bt-3701 and Bacillus megaterium Bm-107].

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    Mu, G; Dong, Y; Huang, G

    1995-10-01

    The results of the interspecific protoplast fusion between B. thuringensis sub. kurstaki Bt-3701 which has pesticide ability, and B. megaterium var. phosphaticum Bm-107 which has decomposing phosphate activity, were reported. High frequency of protoplast formation and regeneration was obtained with 4h activated Bm-107 treated by 100 micrograms/ml lysozyme, and with 2h activated Bt-3701 treated by 3% glycin and mild temperature. Using 40% PEG and 5% nascent Ca2+ to treat the parential protoplast mixture for 3 min at 37 degrees C, 4 stable fusants were obtained. Biological tests show that they have both pesticide ability and decomposing phosphate activity, but which are weaker than that of parential strains. PMID:8701582

  20. Characterization of the penicillin G acylase from Bacillus megaterium ATCC 14945

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    Vanessa Ribeiro de Souza

    2005-06-01

    Full Text Available The purpose of this work was to characterize the enzyme penicillin G acylase (PGA produced by Bacillus megaterium. Purification of the enzyme by ultra/diafiltration did not allow the detection of the PGA band by SDS-PAGE electrophoresis due to the high content of remaining proteins. However, using the DNA of the microorganism, it was possible to replicate the genes of the two B. megaterium PGA reported in literature, showing that the enzyme consisted of two sub-units, having 245 and 537 amino acids each and an average molecular mass of 26950 and 59070 Da, respectively. The parameters studied were: 1 the influence of temperature in the 25-60(0C range, 2 pH in the 5-10 range and 3 substrate concentration, this was tested to obtain results on the Penicillin G hydrolysis reaction rate, using the initial velocities approach. The maximum hydrolysis rate was obtained at 37ºC and pH 8.0. The Michaelis-Menten model fitted well, resulting in estimated Km and Vmax parameters values of 1.83 mM and 0.165*10-3 mmol/min/UI, respectively.O objetivo deste trabalho foi caracterizar a enzima penicilina G acilase (PGA produzida por Bacillus megaterium, uma importante enzima industrial que catalisa a hidrólise de penicilina G, para produção de antibióticos semi-sintéticos. Purificação da enzima por ultra/diafiltração não permitiu detectar a banda de PGA por eletroforese SDS-PAGE devido ao elevado conteúdo de outras proteínas remanescentes. Contudo, utilizando DNA do microrganismo que vem sendo estudado, foi possível amplificar os genes das duas sub-unidades de PGA previstas na literatura, mostrando que a enzima em estudo é também constituída de duas sub-unidades, 245 e 537 aminoácidos cada, com massas moleculares médias de 26950 e 59070 Da, respectivamente. Foram estudadas as influências da temperatura 25-60(0C, pH 5-10, e concentração do substrato na velocidade da reação de hidrólise da penicilina G. A temperatura e pH ótimos foram de 37

  1. A Study of Pt4+-Adsorption and Its Reduction by Bacillus Megaterium D01

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The properties of Pt4+-adsorption and its reduction by Bacillus megaterium D01 were studied by means of ICP,anode-stripping voltammetry,TEM,IR and XPS.The results of ICP analyses showed that the Pt4+-adsorptive efficiency of the strain D01 was as high as 94.3% under the conditions of 100 mg Pt4+/L,1 g biomass/L,pH 3.5 and at 30 ℃ for 24 h.Moreover,it was confirmed from anode stripping voltammetry that the strain D01 possessed a strong reducibility.The TEM analysis indicated that the strain D01 was able to adsorb and reduce Pt4+ to Pt0,small particles.The XPS result further supported the reduction of Pt4+ to Pt2+,followed by the further recuction to Pt0.The IR spectrum implied that D01 biomass adsorption of Pt4+ may result in the complexation of the C=O bond to the Pt species.

  2. Improvement of Ni phytostabilization by inoculation of Ni resistant Bacillus megaterium SR28C.

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    Rajkumar, Mani; Ma, Ying; Freitas, Helena

    2013-10-15

    The use of metal tolerant plants for the phytostabilization of metal contaminated soil is an area of extensive research and development. In this study the effects of inoculation of Ni-resistant bacterial strains on phytostabilization potential of various plants, including Brassica juncea, Luffa cylindrica and Sorghum halepense, were studied. A Ni-resistant bacterial strain SR28C was isolated from a nickel rich serpentine soil and identified as Bacillus megaterium based on the morphological features, biochemical characteristics and partial 16S rDNA sequence analysis. The strain SR28C tolerated concentrations up to 1200 mg Ni L(-1) on a Luria-Bertani (LB) agar medium. Besides, it showed high degree of resistance to various metals (Cu, Zn, Cd, Pb and Cr) and antibiotics (ampicillin, tetracycline, streptomycin, chloramphenicol, penicillin and kanamycin) tested. In addition, the strain bound considerable amounts of Ni in their resting cells. Besides, the strain exhibited the plant growth promoting traits, such as solubilization of phosphate and production of indole-3-acetic acid (IAA) in modified Pikovskayas medium and LB medium, respectively in the absence and presence of Ni. Considering such potential, the effects of SR28C on the growth and Ni accumulation of B. juncea, L. cylindrica and S. halepense, were assessed with different concentrations of Ni in soil. Inoculation of SR28C stimulated the biomass of the test plants grown in both Ni contaminated and non-contaminated soils. Further, SR28C alleviated the detrimental effects of Ni by reducing its uptake and translocation to the plants. This study suggested that the PGPB inoculant due to its intrinsic abilities of growth promotion and attenuation of the toxic effects of Ni could be exploited for phytostabilization of Ni contaminated site.

  3. Use of Streptomyces fradiae and Bacillus megaterium as probiotics in the experimental culture of tiger shrimp Penaeus monodon (Crustacea, Penaeidae

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    Sheikh Aftabuddin

    2013-05-01

    Full Text Available Shrimp hatcheries often beset with diseases, mainly the bacterial infection and antibiotics arewidely used for prevention of disease. Presently, beneficial bacteria (probiotics are used to preventdiseases instead of antibiotics and increasing the production. In the present study, the two new microbialstrains Bacillus megaterium and Streptomyces fradiae isolated from mangrove sediments were applied(experimental culture for the post larval rearing of Penaeus monodon which is compared with controlculture tanks (without probiotics. The water quality condition such as temperature (27-29oC, salinity(26-28‰ and dissolved oxygen (4.7-5.0 mg L-1 of both control and experimental culture were more orless similar. Concentration of ammonia and pH were significantly different (p<0.05 between the controland experimental culture during the study period. The feed assimilation efficiency is higher (above 80%in experimental culture tank when compared to control tank (74.76%. The growth rate was higher -1.70 and 1.67 in S. fradiae, 1.66 and 1.63 in B. megaterium - through feed and water, respectively,while in control tank it was 1.4. The FCR values were 2.06 and 2.12 in S. fradiae treated tanks throughfeed and water, respectively, while 2.51 and 2.55 were observed in B. megaterium treated through feedand water respectively. The FCR value was found higher (4.02 in the control tank. The average totalheterotrophic bacteria (THB and total presumptive Vibrio bacteria both in culture water and post larvaewere lower during experimental culture in compared to control culture. The present study indicates thatthe probiotic treatment using two new microbial strains such as B. megaterium and S. fradiae would helpin better aquaculture production.

  4. Integral Use of Amaranth Starch to Obtain Cyclodextrin Glycosyltransferase, by Bacillus megaterium, to Produce β-Cyclodextrin

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    Arce-Vázquez, María Belem; Ponce-Alquicira, Edith; Delgado-Fornué, Ezequiel; Pedroza-Islas, Ruth; Díaz-Godínez, Gerardo; Soriano-Santos, J.

    2016-01-01

    Cyclodextrin glycosyltransferase (CGTase) is an enzyme that produces cyclodextrins (CDs) from starch and related carbohydrates, producing a mixture of α-, β-, and γ-CDs in different amounts. CGTase production, mainly by Bacillus sp., depends on fermentation conditions such as pH, temperature, concentration of nutrients, carbon and nitrogen sources, among others. Bacillus megaterium CGTase produces those three types of CDs, however, β-CD should prevail. Although, waxy corn starch (CS) is used industrially to obtain CGTase and CDs because of its high amylopectin content, alternative sources such as amaranth starch (AS) could be used to accomplish those purposes. AS has high susceptibility to the amylolytic activity of CGTase because of its 80% amylopectin content. Therefore, the aim of this work was evaluate the AS as carbon source for CGTase production by B. megaterium in a submerged fermentation. Afterwards, the CGTase was purified partially and its activity to synthesize α-, β-, and γ-CDs was evaluated using 1% AS as substrate. B. megaterium produced a 66 kDa CGTase (Topt = 50°C; pHopt = 8.0), from the early exponential growth phase which lasted 36 h. The maximum CGTase specific activity (106.62 ± 8.33 U/mg protein) was obtained after 36 h of culture. CGTase obtained with a Km = 0.152 mM and a Vmax = 13.4 μM/min yielded 40.47% total CDs using AS which was roughly twice as much as that of corn starch (CS; 24.48%). High costs to produce CDs in the pharmaceutical and food industries might be reduced by using AS because of its higher α-, β- and γ-CDs production (12.81, 17.94, and 9.92%, respectively) in a shorter time than that needed for CS. PMID:27721811

  5. CONSTRUCTION OF DOUBLE CHAMBERED MICROBIAL FUEL CELL (MFC USING HOUSEHOLD MATERIALS AND BACILLUS MEGATERIUM ISOLATE FROM TEA GARDEN SOIL

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    Debajit Borah

    2013-08-01

    Full Text Available The current study was carried out for the isolation and screening of potential bioelectricity generating bacteria from tea garden soil samples and also to construct an indigenous microbial fuel cell (MFC using house hold materials. Bacillus megaterium was found to the best isolate for the production of bioelectricity, out of a total of 25 bacterial isolates from soil samples of Lepetkata Tea Estate of Dibrugarh district of Assam. The isolate was identified on the basis of staining techniques and biochemical characteristics. Double chambered MFC was constructed by using two poly acrylic containers of 500 ml volume each. The two chambers were connected using an agar salt bridge and carbon rods were used as electrodes. The electricity generated by the isolate was compared using glucose and fructose as sole carbon source in minimal media. The maximum voltage was found to be 440 mV in presence of glucose as sole carbon source after 84 hrs of incubation at room temperature. The voltage was further increased up to 698 mV after the media was supplemented with 1.5 % (w/v yeast extract, which would have served as additional source of vitamin to the bacteria to proliferate. During the entire study, the experimental set up was allowed to incubate at room temperature and occasional shaking was done manually, hence no external electricity was required. With all the above features the isolate Bacillus megaterium was found to be a good source of bioelectricity.

  6. Decolorization of Textile Dye Vat Blue 66 by Pseudomonas desmolyticum NCIM 2112 and Bacillus megaterium NCIM 2087

    Directory of Open Access Journals (Sweden)

    Archana A. Gurav

    2011-07-01

    Full Text Available The main objective of this investigation is to detoxify the water insoluble dyes like Vat Blue 66 which is other very difficult to decolorize. Wastewater from the textile and other dyestuff industries contain significant amount of synthetic dyes that require prior treatment to prevent groundwater contamination upon being discharged. The microbial decolorization and degradation of dyes has been of considerable interest biological treatment of the wastewater containing dyes. Pseudomonas desmolyticum NCIM 2112 and Bacillus megatherium NCIM 2087 was able to decolorize indole based dye Vat Blue 66 at pH 9, at 25oC. The addition of glucose and yeast extract in the medium enhanced decolorization process. In this investigation, it has been found that the decolorization of this dye was 70% on an average by oxidative degradation by Pseudomonas desmolyticum NCIM 2112 and Bacillus megaterium NCIM 2087. The decolorization was measured as the decrease in absorbance maxima at 530 nm.

  7. Draft Genome Sequence of a Moderately Halophilic Bacillus megaterium Strain, MSP20.1, Isolated from a Saltern of the Little Rann of Kutch, India

    Science.gov (United States)

    Dey, Rinku; Sherathia, Dharmesh; Vanpariya, Sejal; Patel, Ilaxi; Dalsania, Trupti; Savsani, Kinjal; Sukhadiya, Bhoomika; Mandaliya, Mona; Thomas, Manesh; Ghorai, Sucheta; Rupapara, Rupal; Rawal, Priya; Shah, Abhi; Bhayani, Sharmila

    2014-01-01

    The 4.37-Mbp draft genome of a moderately halophilic Bacillus megaterium strain, MSP20.1, isolated from a saltern of the Little Rann of Kutch, India, is reported here. To understand the mechanism(s) of moderate halophilism and to isolate the gene(s) involved in osmotolerance and adaptation, the genome of MSP20.1 was sequenced. PMID:24407642

  8. Biotransformation of colchicinoids into their corresponding 3-O-glucosyl derivatives by selected strains of Bacillus megaterium.

    Science.gov (United States)

    Ponzone, Cesare; Berlanda, Davide; Donzelli, Fabio; Acquati, Valter; Ciulla, Rosalba; Negrini, Alberto; Rovati, Marco; Evangelista, Douglas; Fata, Emilio; Ciceri, Daniele; Perterlongo, Federico; Cabri, Walter

    2014-07-01

    Natural colchicinoids and their semisynthetic derivatives are important active ingredients for pharmaceutical applications. Thiocolchicoside (3-demethoxy-3-glucosyloxythiocolchicine) is used in several countries as standard therapy for the treatment of diseases of the muscle-skeletal system, due to its potent antiinflammatory and myorelaxant properties. Manufacturing of thiocolchicoside requires a key step, the regioselective demethylation and glucosylation of chemically derivative thiocolchicine. High selectivity and efficiency of this transformation cannot be achieved in a satisfactory way with a chemical approach. In particular, the chemical demethylation, a part from requiring toxic and aggressive reagents, generates a complex mixture of products with no industrial usefulness. We report herein an efficient, direct and green biotransformation of thiocolchicine into thiocolchicoside, performed by a specific strain of Bacillus megaterium. The same process, with minor modifications, can be used to convert the by-product 3-O-demethyl-thiocolchicine into thiocolchicoside. In addition, we describe the B. megaterium strain selection process and the best conditions for this effective double biotransformation. The final product has a pharmaceutical quality, and the process has been industrialised. PMID:24553816

  9. Debottlenecking recombinant protein production in Bacillus megaterium under large-scale conditions--targeted precursor feeding designed from metabolomics.

    Science.gov (United States)

    Korneli, Claudia; Bolten, Christoph Josef; Godard, Thibault; Franco-Lara, Ezequiel; Wittmann, Christoph

    2012-06-01

    In the present work the impact of large production scale was investigated for Bacillus megaterium expressing green fluorescent protein (GFP). Specifically designed scale-down studies, mimicking the intermittent and continuous nutrient supply of large- and small-scale processes, were carried out for this purpose. The recombinant strain revealed a 40% reduced GFP yield for the large-scale conditions. In line with extended carbon loss via formation of acetate and carbon dioxide, this indicated obvious limitations in the underlying metabolism of B. megaterium under the large-scale conditions. Quantitative analysis of intracellular amino acids via validated fast filtration protocols revealed that their level strongly differed between the two scenarios. During cultivation in large-scale set-up, the availability of most amino acids, serving as key building blocks of the recombinant protein, was substantially reduced. This was most pronounced for tryptophan, aspartate, histidine, glutamine, and lysine. In contrast alanine was increased, probably related to a bottleneck at the level of pyruvate which also triggered acetate overflow metabolism. The pre-cursor quantifications could then be exploited to verify the presumed bottlenecks and improve recombinant protein production under large-scale conditions. Addition of only 5 mM tryptophan, aspartate, histidine, glutamine, and lysine to the feed solution increased the GFP yield by 100%. This rational concept of driving the lab scale productivity of recombinant microorganisms under suboptimal feeding conditions emulating large scale can easily be extended to other processes and production hosts. PMID:22252649

  10. Development of a biologically based fertilizer, incorporating Bacillus megaterium A6, for improved phosphorus nutrition of oilseed rape.

    Science.gov (United States)

    Hu, Xiaojia; Roberts, Daniel P; Xie, Lihua; Maul, Jude E; Yu, Changbing; Li, Yinshui; Zhang, Shujie; Liao, Xing

    2013-04-01

    Sustainable methods with diminished impact on the environment need to be developed for the production of oilseed rape in China and other regions of the world. A biological fertilizer consisting of Bacillus megaterium A6 cultured on oilseed rape meal improved oilseed rape seed yield (P rape meal without strain A6 in 1 of 2 experiments, suggesting a role for strain A6 in improving yield. Strain A6 was capable of solubilizing phosphorus from rock phosphate in liquid culture and produced enzymes capable of mineralizing organic phosphorus (acid phosphatase, phytase) in liquid culture and in the biological fertilizer. The biologically based fertilizer, containing strain A6, improved plant phosphorus nutrition in greenhouse pot experiments resulting in significantly greater available phosphorus in natural soil and in significantly greater plant phosphorus content relative to the nontreated control. Seed yield and available phosphorus in natural soil were significantly greater with a synthetic chemical fertilizer treatment, reduced in phosphorus content, than the biological fertilizer treatment, but a treatment containing the biological fertilizer combined with the synthetic fertilizer provided the significantly greatest seed yield, available phosphorus in natural soil, and plant phosphorus content. These results suggest that the biological fertilizer was capable of improving oilseed rape seed yield, at least in part, through the phosphorus-solubilizing activity of B. megaterium A6. PMID:23586746

  11. Identification of lipopeptides in Bacillus megaterium by two-step ultrafiltration and LC-ESI-MS/MS.

    Science.gov (United States)

    Ma, Yunxiao; Kong, Qing; Qin, Chong; Chen, Yulin; Chen, Yujie; Lv, Ruihuan; Zhou, Guanghui

    2016-12-01

    A two-step ultrafiltration and liquid chromatography-electronic spray ionization-tandem mass spectrometry (LC-ESI-MS/MS) were applied to investigate the nature of lipopeptides in the methanol extract harvested from marine Bacillus megaterium. The structure of lipopeptide homologs were characterized by using collision-induced dissociation mass spectrometry analysis. Collectively, two kinds of linear fengycin A and B with a double bond were characterized for the first time besides the popular cyclic fengycin A and B with the difference in the amino acid at position 6 of the peptide moiety. It is worth noting that two kinds of linear isoforms of surfactins and two kinds of cyclic isoforms of esperin with a smaller peptide ring formed by the fifth L-Asp and β-hydroxy fatty acid were also separated and identified in addition to the all seven kinds of cyclic isoforms of surfactins. Only one analogue (bacillomycin D) of iturin family was identified in this research. To our best knowledge, this is the first report that more than 40 variants of lipopeptides from one strain of Bacillus were identified using our potent purification and identification methods. PMID:27639854

  12. Purification and characterization of a protease produced by Bacillus megaterium RRM2: application in detergent and dehairing industries.

    Science.gov (United States)

    Rajkumar, Renganathan; Jayappriyan, Kothilmozhian Ranishree; Rengasamy, Ramasamy

    2011-12-01

    An alkaline serine protease produced by Bacillus megaterium RRM2 isolated from the red alga, Kappaphycus alvarezii (Doty) Doty ex Silva was studied for the first time and the same analyzed for the production of protease in the present study. Identification of the bacterium was done on the basis of both biochemical analysis and by 16S rDNA sequence analysis. The extracellular protease obtained from B. megaterium RRM2 was purified by a three-step process involving ammonium sulphate precipitation, gel filtration (Sephadex G100) and Q-Sepharose column chromatography. The purity was found to be 30.6-fold with a specific activity of 3591.5 U/mg protein with a molecular weight of 27 kDa. The metal ions Ca(2+), Mg(2+), K(+) and Na(+) marginally enhanced the activity of the purified enzyme while Hg(2+), Cu(2+), Fe(2+), CO(2+) and Zn(2+), had reduced the activity. The enzyme was found to be active in the pH range of 9.0-10.0 and remained active up to 60 °C. Phenyl Methyl Sulfonyl Fluoride (PMSF) inhibited the enzyme activity, thus, confirming that this enzyme is an alkaline serine protease. Likewise, DTT also inhibited the enzyme thus confirming the disulfide nature of the enzyme. The enzyme exhibited a high degree of tolerance to Sodium Dodecyl Sulphate (SDS). The partially purified protease when used as an additive in the commercial detergents was found to be a suitable source for washing clothes especially those stained with blood. Further, it showed good dehairing activity within a short duration in goat skin without affecting its collagen component. PMID:21780140

  13. A native plant growth promoting bacterium, Bacillus megaterium B55, rescues growth performance of an ethylene insensitive plant genotype in nature

    OpenAIRE

    Dorothea Gertrud Meldau; Hoang Hoa Long; Ian Thomas Baldwin

    2012-01-01

    Many plants have intimate relationships with soil microbes that through a variety of mechanisms improve the plant’s growth and fitness. Bacillus megaterium is a natural endophyte isolated from Nicotiana attenuata plant roots growing in native soils. A particular isolate (B55), was found to have dramatic plant growth promoting (PGP) effects on wild type (WT) and transgenic plants impaired in ethylene (ET) perception (35S-etr1), the genotype from which this bacteria was first isolated. B55 not ...

  14. The Identification on a Strain of Bacillus Megaterium%一株巨大芽孢杆菌的分类鉴定

    Institute of Scientific and Technical Information of China (English)

    詹建立; 周娜

    2014-01-01

    基于16S rRNA序列分析和生理生化实验,对从伽师瓜病瓜上分离的一株细菌ZP-1进行了分类鉴定.结果表明,该分离菌株ZP-1与Bacillus megaterium strain PAB1C5(EU221343.1)具有99%的相似性.因此,初步推断ZP-1菌株隶属于巨大芽孢杆菌属(Bacillus magaterium).%Based on 16S rRNA sequence analysis and physiological and biochemical experiment, it isolated and i-dentified a strain of ZP-1bacterium from Jiashi muskmelon. It showed that the isolate shared the 99% similarity with Bacillus megaterium strain PAB1C5 (EU221343.1). And it was deduced that the isolate belonged to Bacillus magaterium. So, it would provide the strain resource and the theoretical data for the application of Bacillus maga-terium.

  15. Rhizobacterial Strain Bacillus megaterium BOFC15 Induces Cellular Polyamine Changes that Improve Plant Growth and Drought Resistance.

    Science.gov (United States)

    Zhou, Cheng; Ma, Zhongyou; Zhu, Lin; Xiao, Xin; Xie, Yue; Zhu, Jian; Wang, Jianfei

    2016-01-01

    Plant-growth-promoting rhizobacteria can improve plant growth, development, and stress adaptation. However, the underlying mechanisms are still largely unclear. We investigated the effects of Bacillus megaterium BOFC15 on Arabidopsis plants. BOFC15 produced and secreted spermidine (Spd), a type of polyamine (PA) that plays an important role in plant growth. Moreover, BOFC15 induced changes in the cellular PAs of plants that promoted an increase of free Spd and spermine levels. However, these effects were remarkably abolished by the addition of dicyclohexylamine (DCHA), a Spd biosynthetic inhibitor. Additionally, the inoculation with BOFC15 remarkably increased plant biomass, improved root system architecture, and augmented photosynthetic capacity. Inoculated plants also displayed stronger ability to tolerate drought stress than non-inoculated (control) plants. Abscisic acid (ABA) content was notably higher in the inoculated plants than in the control plants under drought stress and polyethylene glycol (PEG)-induced stress conditions. However, the BOFC15-induced ABA synthesis was markedly inhibited by DCHA. Thus, microbial Spd participated in the modulation of the ABA levels. The Spd-producing BOFC15 improved plant drought tolerance, which was associated with altered cellular ABA levels and activated adaptive responses. PMID:27338359

  16. Rhizobacterial Strain Bacillus megaterium BOFC15 Induces Cellular Polyamine Changes that Improve Plant Growth and Drought Resistance

    Directory of Open Access Journals (Sweden)

    Cheng Zhou

    2016-06-01

    Full Text Available Plant-growth-promoting rhizobacteria can improve plant growth, development, and stress adaptation. However, the underlying mechanisms are still largely unclear. We investigated the effects of Bacillus megaterium BOFC15 on Arabidopsis plants. BOFC15 produced and secreted spermidine (Spd, a type of polyamine (PA that plays an important role in plant growth. Moreover, BOFC15 induced changes in the cellular PAs of plants that promoted an increase of free Spd and spermine levels. However, these effects were remarkably abolished by the addition of dicyclohexylamine (DCHA, a Spd biosynthetic inhibitor. Additionally, the inoculation with BOFC15 remarkably increased plant biomass, improved root system architecture, and augmented photosynthetic capacity. Inoculated plants also displayed stronger ability to tolerate drought stress than non-inoculated (control plants. Abscisic acid (ABA content was notably higher in the inoculated plants than in the control plants under drought stress and polyethylene glycol (PEG-induced stress conditions. However, the BOFC15-induced ABA synthesis was markedly inhibited by DCHA. Thus, microbial Spd participated in the modulation of the ABA levels. The Spd-producing BOFC15 improved plant drought tolerance, which was associated with altered cellular ABA levels and activated adaptive responses.

  17. Rhizobacterial Strain Bacillus megaterium BOFC15 Induces Cellular Polyamine Changes that Improve Plant Growth and Drought Resistance

    Science.gov (United States)

    Zhou, Cheng; Ma, Zhongyou; Zhu, Lin; Xiao, Xin; Xie, Yue; Zhu, Jian; Wang, Jianfei

    2016-01-01

    Plant-growth-promoting rhizobacteria can improve plant growth, development, and stress adaptation. However, the underlying mechanisms are still largely unclear. We investigated the effects of Bacillus megaterium BOFC15 on Arabidopsis plants. BOFC15 produced and secreted spermidine (Spd), a type of polyamine (PA) that plays an important role in plant growth. Moreover, BOFC15 induced changes in the cellular PAs of plants that promoted an increase of free Spd and spermine levels. However, these effects were remarkably abolished by the addition of dicyclohexylamine (DCHA), a Spd biosynthetic inhibitor. Additionally, the inoculation with BOFC15 remarkably increased plant biomass, improved root system architecture, and augmented photosynthetic capacity. Inoculated plants also displayed stronger ability to tolerate drought stress than non-inoculated (control) plants. Abscisic acid (ABA) content was notably higher in the inoculated plants than in the control plants under drought stress and polyethylene glycol (PEG)-induced stress conditions. However, the BOFC15-induced ABA synthesis was markedly inhibited by DCHA. Thus, microbial Spd participated in the modulation of the ABA levels. The Spd-producing BOFC15 improved plant drought tolerance, which was associated with altered cellular ABA levels and activated adaptive responses. PMID:27338359

  18. Alleviating salt stress in tomato seedlings using Arthrobacter and Bacillus megaterium isolated from the rhizosphere of wild plants grown on saline-alkaline lands.

    Science.gov (United States)

    Fan, Pengfei; Chen, Daitao; He, Yanan; Zhou, Qingxia; Tian, Yongqiang; Gao, Lihong

    2016-11-01

    Salt-induced soil degradation is common in farmlands and limits the growth and development of numerous crop plants in the world. In this study, we isolated salt-tolerant bacteria from the rhizosphere of Tamarix chinensis, Suaeda salsa and Zoysia sinica, which are common wild plants grown on a saline-alkaline land, to test these bacteria's efficiency in alleviating salt stress in tomato plants. We screened out seven strains (TF1-7) that are efficient in reducing salt stress in tomato seedlings. The sequence data of 16S rRNA genes showed that these strains belong to Arthrobacter and Bacillus megaterium. All strains could hydrolyze casein and solubilize phosphate, and showed at least one plant growth promotion (PGP)-related gene, indicating their potential in promoting plant growth. The Arthrobacter strains TF1 and TF7 and the Bacillus megaterium strain TF2 and TF3 could produce indole acetic acid under salt stress, further demonstrating their PGP potential. Tomato seed germination, seedling length, vigor index, and plant fresh and dry weight were enhanced by inoculation of Arthrobacter and B. megaterium strains under salt stress. Our results demonstrated that salt-tolerant bacteria isolated from the rhizosphere of wild plants grown on saline-alkaline lands could be used for alleviating salt stress in crop plants. PMID:27196364

  19. Wheat bran as a substrate for thermo stable alpha-amylase production by gamma irradiated bacillus megaterium in solid state fermentation

    International Nuclear Information System (INIS)

    Thermo stable alpha-amylase (EC 3.2.1.1) production from cheap agriculture-industrial waste wheat bran (WB) medium by superior potent gamma irradiated locally isolated strain of Bacillus megaterium in solid state fermentation (SSF) was studied. A highly yielding, stable enhanced isolated strain of bacillus megaterium in solid state fermentation (SSF) was studied. A highly yielding stable enhanced isolate B. megaterium- gamma 21F derived from the 10 kGy, treatment, exhibited the highest alpha-amylase activity under SSF, with 2.8 fold more enzyme titer as compared to the unirradiated wild strain. A vancomycin (Vm) resistant gamma irradiated enhanced isolate B. megaterium-gamma 21F2 (which was selected throughout the subsequent work) secreted (1.27 and 3.58) folds superior titers of alpha-amylase than the gamma irradiated parent isolate (B.megaterium -gamma21F) and unirradiated wild strain, respectively under SSF process. The effects of various parameters, such as moistening agent, initial moisture content level, initial ph, incubation temperature, inoculum size and incubation time on thermo stable alpha-amylase production by B.megaterium-gamma 21F2 under SSF were studied. Maximum enzyme production was recorded in WB medium moistened with (1:2, w/v) distilled water at initial ph (7.0) and inoculated with (2.24 x 108 cells/g WB) after 48 h incubation at 40 C degree. Between different solvents used for enzyme extraction from fermented WB mass, distilled water at ph (7.0) was the superior efficient leaching solvent. The specific activity of the precipitated partially purified crude thermo stable enzyme was (258.7 U/mg protein) with ph optima (6.5-7.0), at optimal temperatures (65-70 c degree) and it retained about 53% of its maximum activity after 12 h incubation at 70 c degree. The partially purified crude enzyme was used for starch digestion (5%0 under optimized reaction conditions, wherein (98.2%) starch hydrolysis was attained after 6 h

  20. Isolation, purification and characterisation of an organic solvent-tolerant Ca2+-dependent protease from Bacillus megaterium AU02.

    Science.gov (United States)

    Priya, J Deepa Arul; Divakar, K; Prabha, M Suriya; Selvam, G Panneer; Gautam, Pennathur

    2014-01-01

    A new organic solvent-tolerant strain Bacillus megaterium AU02 which secretes an organic solvent-tolerant protease was isolated from milk industry waste. Statistical methods were employed to achieve optimum protease production of 43.6 U/ml in shake flask cultures. The productivity of the protease was increased to 53 U/ml when cultivated under controlled conditions in a 7-L fermentor. The protease was purified to homogeneity by a three-step process with 24 % yield and specific activity of 5,375 U/mg. The molecular mass of the protease was found to be 59 kDa. The enzyme was active over a wide range of pH (6.0–9.0), with an optimum activity at pH 7.0 and temperature from 40 to 70 °C having an optimum activity at 50 °C. The thermal stability of the enzyme increased significantly in the presence of CaCl2, and it retained 90 % activity at 50 °C for 3 h. The Km and Vmax values were determined as 0.722 mg/ml and 0.018 U/mg respectively. The metalloprotease exhibited significant stability in the presence of organic solvents with log P values more than 2.5, nonionic detergents and oxidising agent. An attempt was made to test the synthesis of aspartame precursor (Cbz-Asp-Phe-NH2) which was catalysed by AU02 protease in the presence of 50 % DMSO. These properties of AU02 protease make it an ideal choice for enzymatic peptide synthesis in organic media. PMID:24122712

  1. Microbial induced corrosion (MIC) on DHP copper by Desulfovibrio desulfuricans and Bacillus megaterium strains in media simulating heater waters

    International Nuclear Information System (INIS)

    The complexity and diversity of microbial populations in water heating systems of steam generators make it necessary to study the magnitude of the metabolic activity of bacteria and biofilm development that may lead to degradation of metal components through microbial induced corrosion (MIC). Electrolytes simulating the conditions found in heater water networks were used to induce biofilm formation on DHP copper coupons by Desulfovibrio desulfuricans DSMZ and Bacillus megaterium C10, a commercial strain and an isolate from these waters, respectively. In order to enhance their action, industrial waters enriched with the minimum nutrient content such as sodium lactate and sodium sulphite for the DSMZ strain and glucose, proteose peptone and starch for the C10 strain were employed. Biofilm formation was studied under controlled temperature, time, shaking, pH and concentrations of the media used in this study. Then, the samples were electrochemically tested in an artificial solution of sea water as control medium, based on the hypothesis that the action of an aggressive biofilm/electrolyte medium generates damaged and non-damaged areas on the metal surface, and assuming that the sea water trial can detect the latter. Hence, a higher anodic current was associated with a lower degradation of the metal surface by the action of one of the media under study. All these trials were performed along with bacterial count, scanning electron microscopy (SEM) and atomic absorption spectroscopy (AAS). Furthermore, it was possible to identify under which conditions MIC on DHP copper occurred and complex mechanisms from retention of cations to diffusion processes at the biofilm/tested media interface level were proposed. Surface corrosion by MIC took place on DHP copper; therefore, greater control on the treatment of industrial waters is highly desirable. (author)

  2. Microbial induced corrosion (MIC) on DHP copper by Desulfovibrio desulfuricans and Bacillus megaterium strains in media simulating heater waters

    Energy Technology Data Exchange (ETDEWEB)

    Zumelzu, E.; Cabezas, C.; Schoebitz, R.; Ugarte, R.; Rodriguez, E.D.; Rios, J. [Universidad Austral de Chile, Valdivia (Chile)

    2003-01-01

    The complexity and diversity of microbial populations in water heating systems of steam generators make it necessary to study the magnitude of the metabolic activity of bacteria and biofilm development that may lead to degradation of metal components through microbial induced corrosion (MIC). Electrolytes simulating the conditions found in heater water networks were used to induce biofilm formation on DHP copper coupons by Desulfovibrio desulfuricans DSMZ and Bacillus megaterium C10, a commercial strain and an isolate from these waters, respectively. In order to enhance their action, industrial waters enriched with the minimum nutrient content such as sodium lactate and sodium sulphite for the DSMZ strain and glucose, proteose peptone and starch for the C10 strain were employed. Biofilm formation was studied under controlled temperature, time, shaking, pH and concentrations of the media used in this study. Then, the samples were electrochemically tested in an artificial solution of sea water as control medium, based on the hypothesis that the action of an aggressive biofilm/electrolyte medium generates damaged and non-damaged areas on the metal surface, and assuming that the sea water trial can detect the latter. Hence, a higher anodic current was associated with a lower degradation of the metal surface by the action of one of the media under study. All these trials were performed along with bacterial count, scanning electron microscopy (SEM) and atomic absorption spectroscopy (AAS). Furthermore, it was possible to identify under which conditions MIC on DHP copper occurred and complex mechanisms from retention of cations to diffusion processes at the biofilm/tested media interface level were proposed. Surface corrosion by MIC took place on DHP copper; therefore, greater control on the treatment of industrial waters is highly desirable. (author)

  3. Evaluation of children as sources of bioaerosols in a climate chamber study. [Staphylococcus epidermidis; Saprophyticus; Bacillus spp. ; Bacillus megaterium; Acinetobacter spp

    Energy Technology Data Exchange (ETDEWEB)

    Lundqvist, G.R.; Aalykke, C.; Bonde, G.J. (Univ. of Aarhus (Denmark))

    1990-01-01

    Emissions of viable particles from a group of children were measured under controlled conditions in a climate chamber that simulated indoor environmental exposure in day-care institutions with tight building envelopes and outdoor air supply by natural infiltration only. Bioaerosol sampling was simultaneously applied with slit samplers and sediment plates. A total of 142 strains was identified. Most of these were from sediment plates (95%) as the colonies on the slit sampler were more crowded and too confluent for separation. On sediment plates, coryneform bacteria dominated (27-85%), followed in frequency by micrococci (4-50%), Staphylococcus epidermidis and saprophyticus (12-43%), Bacillus spp., most frequently B. megaterium (12-33%), and Acinetobacter spp. (11-14%). From the slit sampler plates, staphylococci dominated (67%), followed by coryneform species and micrococci (17%). Within the first hour after the group left the chamber, the number of colony forming units (CFU) suspended in the air decreased, corresponding to an equivalent dilution ventilation rate of 2.0 ACH (air changes per hour) for bacteria and 1.7 ACH for mold spores due to the catching of particles on surfaces and to die away of viable microorganisms. Accordingly, microbial surface contamination revealed an increase at the same time.

  4. The self-assembly of a cyclic lipopeptides mixture secreted by a B. megaterium strain and its implications on activity against a sensitive Bacillus species.

    Directory of Open Access Journals (Sweden)

    Manuel T Pueyo

    Full Text Available Cyclic lipopeptides are produced by a soil Bacillus megaterium strain and several other Bacillus species. In this work, they are detected both in the Bacillus intact cells and the cells culture medium by MALDI-TOF mass spectrometry. The cyclic lipopeptides self-assemble in water media producing negatively charged and large aggregates (300-800 nm of mean hydrodynamic radius as evaluated by dynamic light scattering and zeta-potential analysis. The aggregate size depends on pH and ionic strength. However, it is not affected by changes in the osmolarity of the outer medium suggesting the absence of an internal aqueous compartment despite the occurrence of low molecular weight phospholipids in their composition as determined from inorganic phosphorus analysis. The activity against a sensitive Bacillus cereus strain was evaluated from inhibition halos and B. cereus lysis. Essential features determining the antibiotic activity on susceptible Bacillus cereus cells are the preserved cyclic moiety conferring cyclic lipopeptides resistance to proteases and the medium pH. The aggregates are inactive per se at the pH of the culture medium which is around 6 or below. The knock out of the sensitive cells only takes place when the aggregates are disassembled due to a high negative charge at pH above 6.

  5. Purification and Properties of a New L-Sorbose Dehydrogenase Accelerative Protein from Bacillus megaterium Bred by Ion-Beam Implantation

    Institute of Scientific and Technical Information of China (English)

    ZHAO Shiguang; YAO Liming; SU Caixin; WANG Wao; WANG Jun; TANG Mingli; YU Zengliang

    2008-01-01

    Bacillus megaterium BM302 bred by ion-beam implantation produces L-sorbose dehydrogenase accelerative protein (SAP) to accelerate the activity of L-sorbose dehydrogenase (SDH) of Gluconobacter oxydans in the 2-keto-L-gulonic acid (2KLG) fermentation from L-sorbose by the mixed culture of B. megaterium BM302 and G. oxydans. The SAP purified by three chromatographic steps gave 35-fold purification with a yield of 13% and a specific activity of 5.21 units/mg protein. The molecular weight of the purified SAP was about 58 kDa. The SDH accelerative activity of SAP at pH 7 and 50℃ was the highest. Additionally, it retained 60% activity at a pH range of 6.5 ~ 10 and was stable at 20℃~ 60℃. After 0.32-unit SAP was added to the single cultured G. oxydans strains, the SDH activity was apparently accelerated and the 2KLG yield of GO29, GO112, G0and GI13 was enhanced 2.1, 3.3, 3.5 and 2.9 folds respectively over that of the strains without the addition of SAP.

  6. Application of Escherichia coli phage K1E DNA-dependent RNA polymerase for in vitro RNA synthesis and in vivo protein production in Bacillus megaterium.

    Science.gov (United States)

    Stammen, Simon; Schuller, Franziska; Dietrich, Sylvia; Gamer, Martin; Biedendieck, Rebekka; Jahn, Dieter

    2010-09-01

    Gene "7" of Escherichia coli phage K1E was proposed to encode a novel DNA-dependent RNA polymerase (RNAP). The corresponding protein was produced recombinantly, purified to apparent homogeneity via affinity chromatography, and successfully employed for in vitro RNA synthesis. Optimal assay conditions (pH 8, 37 degrees C, 10 mM magnesium chloride and 1.3 mM spermidine) were established. The corresponding promoter regions were identified on the phage genome and summarized in a sequence logo. Surprisingly, next to K1E promoters, the SP6 promoter was also recognized efficiently in vitro by K1E RNAP, while the T7 RNAP promoter was not recognized at all. Based on these results, a system for high-yield in vitro RNA synthesis using K1E RNAP was established. The template plasmid is a pUC18 derivative, which enables blue/white screening for positive cloning of the target DNA. Production of more than 5 microg of purified RNA per microgram plasmid DNA was achieved. Finally, in vivo protein production systems for Bacillus megaterium were established based on K1E and SP6 phage RNAP transcription. Up to 61.4 mg g (CDW) (-1) (K1E RNAP) of the reporter protein Gfp was produced in shaking flask cultures of B. megaterium. PMID:20596705

  7. Process development for the production of 15β-hydroxycyproterone acetate using Bacillus megaterium expressing CYP106A2 as whole-cell biocatalyst

    DEFF Research Database (Denmark)

    Kiss, Flora M.; Lundemo, Marie Therese; Zapp, Josef;

    2015-01-01

    for this biocatalyst. It is highly selective towards the 15β position, but the 6 β, 7 a/β, 9a, 11a and 15a positions have also been described as targets for hydroxylation. Based on the broad substrate spectrum and hydroxylating capacity, it is an excellent candidate for the production of human drug metabolites...... and drug precursors. Results: In this work, we demonstrate the conversion of a synthetic testosterone derivative, cyproterone acetate, by CYP106A2 under in vitro and in vivo conditions. Using a Bacillus megaterium whole-cell system overexpressing CYP106A2, sufficient amounts of product for structure...... titers for future industrial application, the main bottlenecks of the reaction were addressed. Using 2-hydroxypropyl-β-cyclodextrin, an effective bioconversion of 98% was achieved using 1 mM substrate concentration, corresponding to a product formation of 0.43 g/L, at a 400 mL scale. Conclusions: Here we...

  8. Mechanism of Solubilizing Phosphate by Bacillus megaterium%解磷巨大芽孢杆菌(Bacillus megaterium)的溶磷机理探讨

    Institute of Scientific and Technical Information of China (English)

    吴海燕; 金荣德; 范作伟; 胡会军; 孙甜田; 赵兰坡

    2014-01-01

    One strain of bacterium with strong phosphate solubilizing activity was isolated from phosphorus deficiency black soil samples in corn belt of the Songliao plain of Northeast China and identified as Bacil-lus megaterium by 1 6s rRNA gene sequencing .The soluble-P concentration continuously increased during the incubation periods and the total amount of soluble P released in culture filtrate was detected at 420 mg/kg after 4 days of inoculation .The particle size of calcium phosphorus surface was found to be destroyed and the size changed between 77.5-1 24.2 nm,while no changes in the surface and size were found in the control with the size of 1 32.7-1 81.7 nm .The metabolites of phosphate solubilizing bacteria fermentation broth were analyzed by high-performance liquid chromatograph (HPLC)and Fourier trans-form infrared spectrometer (FTIR)to determine the final fermentation products of phosphorus solubilizing bacteria for gluconic acid .%从松辽平原玉米带中部的公主岭市贫磷黑土中分离筛选了1株具有较强溶磷能力的菌株,并通过16S rRNA基因测序鉴定为解磷巨大芽孢杆菌(Bacillus megaterium)。测定了溶磷圈和发酵液有效磷质量分数,探讨了菌株溶磷机理。结果表明:该菌株在以磷酸钙为唯一磷源的培养基上,能够形成明显的溶磷圈;发酵培养到第4天,发酵液的有效磷质量分数达到420 mg/kg。通过粒子大小分析仪,测定了溶磷菌发酵液中的磷酸钙颗粒大小,对照处理的磷酸钙粒径为132.7~181.7 nm,溶磷菌液处理的磷酸钙粒径为77.5~124.2 nm,磷酸钙颗粒明显变小;通过高效液相色谱法(HPLC)和傅里叶变换红外光谱分析仪(FTIR),对溶磷菌发酵液代谢产物进行分析,确定了溶磷菌最终的发酵产物为葡萄糖酸。

  9. Evidence against the Bm1P1 protein as a positive transcription factor for barbiturate-mediated induction of cytochrome P450BM-1 in bacillus megaterium.

    Science.gov (United States)

    Shaw, G C; Sung, C C; Liu, C H; Lin, C H

    1998-04-01

    The Bm1P1 protein was previously proposed to act as a positive transcription factor involved in barbiturate-mediated induction of cytochrome P450BM-1 in Bacillus megaterium. We now report that the bm1P1 gene encodes a protein of 217 amino acids, rather than the 98 amino acids as reported previously. In vitro gel shift assays indicate that the Bm1P1 protein did not interact with probes comprising the regulatory regions of the P450BM-1 gene. Moreover, disruption of the bm1P1 gene did not markedly affect barbiturate induction of P450BM-1 expression. A multicopy plasmid harboring only the P450BM-1 promoter region could increase expression of the chromosome-encoded P450BM-1. The level of expression is comparable with that shown by a multicopy plasmid harboring the P450BM-1 promoter region along with the bm1P1 gene. These results strongly suggest that the Bm1P1 protein is unlikely to act as a positive regulator for barbiturate induction of P450BM-1 expression. Finally, deletion of the Barbie box did not markedly diminish the effect of pentobarbital on expression of a reporter gene transcriptionally fused to the P450BM-1 promoter. This suggests that the Barbie box is unlikely to be a key element in barbiturate-mediated induction of P450BM-1. PMID:9525898

  10. Effect of biosurfactant and fertilizer on biodegradation of crude oil by marine isolates of Bacillus megaterium, Corynebacterium kutscheri and Pseudomonas aeruginosa.

    Science.gov (United States)

    Thavasi, Rengathavasi; Jayalakshmi, Singaram; Banat, Ibrahim M

    2011-01-01

    This study was conducted to investigate the effects of fertilizers and biosurfactants on biodegradation of crude oil by three marine bacterial isolates; Bacillus megaterium, Corynebacterium kutscheri and Pseudomonas aeruginosa. Five sets of experiments were carried out in shake flask and microcosm conditions with crude oil as follows: Set 1-only bacterial cells added (no fertilizer and biosurfactant), Set 2-with additional fertilizer only, Set 3-with additional biosurfactant only, Set 4-with added biosurfactant+fertilizer, Set 5-with no bacterial cells added (control), all the above experimental sets were incubated for 168 h. The biosurfactant+fertilizer added Set 4, resulted in maximum crude oil degradation within shake flask and microcosm conditions. Among the three bacterial isolates, P. aeruginosa and biosurfactant produced by this strain resulted in maximum crude oil degradation compared to the other two bacterial strains investigated. Interestingly, when biosurfactant and bacterial cells were used (Set 3), significant oil biodegradation activity occurred and the difference between this treatment and that in Set 4 with added fertilizer+biosurfactant were only 4-5% higher degradation level in shake flask and 3.2-7% in microcosm experiments for all three bacterial strains used. It is concluded that, biosurfactants alone capable of promoting biodegradation to a large extent without added fertilizers, which will reduce the cost of bioremediation process and minimizes the dilution or wash away problems encountered when water soluble fertilizers used during bioremediation of aquatic environments. PMID:20863694

  11. Comparative proteomic analysis of high cell density cultivations with two recombinant Bacillus megaterium strains for the production of a heterologous dextransucrase

    Directory of Open Access Journals (Sweden)

    Deckwer Wolf-Dieter

    2006-10-01

    Full Text Available Abstract High cell density cultivations were performed under identical conditions for two Bacillus megaterium strains (MS941 and WH320, both carrying a heterologous dextransucrase (dsrS gene under the control of the xylA promoter. At characteristic points of the cultivations (end of batch, initial feeding, before and after induction the proteome was analyzed based on two dimensional gel electrophoresis and mass spectrometric protein identification using the protein database "bmegMEC.v2" recently made available. High expression but no secretion of DsrS was found for the chemical mutant WH320 whereas for MS 941, a defined protease deficient mutant of the same parent strain (DSM319, not even expression of DsrS could be detected. The proteomic analysis resulted in the identification of proteins involved in different cellular pathways such as in central carbon and overflow metabolism, in protein synthesis, protein secretion and degradation, in cell wall metabolism, in cell division and sporulation, in membrane transport and in stress responses. The two strains exhibited considerable variations in expression levels of specific proteins during the different phases of the cultivation process, whereas induction of DsrS production had, in general, little effect. The largely differing behaviour of the two strains with regard to DsrS expression can be attributed, at least in part, to changes observed in the proteome which predominantly concern biosynthetic enzymes and proteins belonging to the membrane translocation system, which were strongly down-regulated at high cell densities in MS941 compared with WH320. At the same time a cell envelope-associated quality control protease and two peptidoglycan-binding proteins related to cell wall turnover were strongly expressed in MS941 but not found in WH320. However, to further explain the very different physiological responses of the two strains to the same cultivation conditions, it is necessary to identify the

  12. 巨大芽孢杆菌Bacillus megaterium NCIB8508蔗糖磷酸化酶生物催化合成α-熊果苷的研究%Biocatalytic Synthesis of α-Arbutin by Bacillus Megaterium NCIB 8508 Sucrose Phosphorylase

    Institute of Scientific and Technical Information of China (English)

    李群良; 张欣英; 杨克迪; 姚评佳; 魏远安

    2011-01-01

    利用巨大芽孢杆菌Bacillus megaterium NCIB 8508蔗糖磷酸化醇生物催化合成α-熊果苷.考察了缓冲溶液pH值、缓冲溶液浓度、对苯二酚浓度、反应物摩尔比及反应时间等因素对反应的影响.结果表明,以湿菌体50 mg·mL-1破碎细胞粗酶液催化合成α-熊果苷,在pH值为6.5的30 mmol·L-1磷酸盐缓冲溶液中,温度为30℃、摇床转速为180 r·min-1、对苯二酚浓度为5 mmo1·L-1、对苯二酚与蔗糖摩尔比为1:5、反应时间为24 h的优化条件下,a-熊果苷含量达0.35 mg·mL-1、对苯二酚选择性为4.0%、对苯二酚转化率为74%.%The biocatalytic synthesis of α-arbutin by Bacillus megaterium NCIB 8508 sucrose phosphorylase was studied in this paper. The effects of pH value of buffer solution, buffer solution concentration, hydroquinone concentration, reactant molar ratio, and reaction time were investigated. The results showed that with crude enzyme from 50 mg·mL-1 wet cells, α-arbutin was synthesized under the optimized reaction conditions as follows:buffer solution pH value of 6.5, buffer solution concentration of 30 mmol · L-1, temperature of 30℃, shaking speed of 180 r · min-1 , hydroquinone concentration of 5 mmol·L-1, molar ratio of hydroquinone to sucrose of 1 : 5, and reaction time of 24 h, the α-arbutin content was 0.35 mg·mL-1 , the selectivity of hydroquinone was 4.0%, and the conversion rate of hydroquinone was 74%.

  13. The effects of inoculation with phosphate solubilizing bacteria Bacillus megaterium C4 in the AM fungal hyphosphere on soil organic phosphorus mineralization and plant uptake%菌丝室接种解磷细菌Bacillus megaterium C4对土壤有机磷矿化和植物吸收的影响

    Institute of Scientific and Technical Information of China (English)

    张林; 丁效东; 王菲; 田芷源; 冯固

    2012-01-01

    通过30μm尼龙网将根盒分成根室和菌丝室,菌丝室中的低磷土壤施加75 mg P/kg土壤的植酸钙,研究了菌丝室土壤中丛枝菌根(AM)真菌Glomus intraradices和解磷细菌Bacillus megaterium C4对有机磷的矿化和吸收.结果表明,在试验条件下,植酸钙的溶解性很低,对土壤溶液有机磷的贡献不大.接种解磷细菌C4提高了土壤中磷酸酶的活性,减少了土壤中有机磷的含量.但是,由于存在解磷细菌与AM真菌对磷的竞争,解磷细菌矿化出的磷大部分被自身利用,AM真菌的生长受到抑制,解磷细菌对植物磷营养的改善没有表现出显著的贡献.

  14. 基于文献挖掘的巨大芽胞杆菌代谢网络模型的构建与分析%Reconstruction and analysis of Bacillus megaterium metabolic model based on literature study

    Institute of Scientific and Technical Information of China (English)

    周冒达; 邹伟; 刘立明; 陈坚

    2012-01-01

    [ Objective] The aim of this study is to reconstruct the metabolic model of Bacillus megaterium by literature study, which would be used to elucidate physiological properties in detail and refine physiological functions. [Methods] We built a literature database by searching B. Megaterium related literatures from Web of Science, PubMed, United States Patent and Trademark Office ( USPTO) Patent Databases, Derwent Innovations Index and China National Knowledge Infrastructure ( CNKI). Depending on this database, we extracted the functional genes, enzymes, metabolites and metabolic reactions (including transport reactions) through literature studies. Then, we filled gaps through KECC Mapper and adding sink reactions. The operation of Matlab programs reconstructed the metabolic model ( in the form of Systems Biology Markup Language) of B. Megaterium. [Results] The refined metabolic network model contained 292 metabolic reactions, 378 metabolites, 220 enzymes and 217 genes. With the restrictive condition of 1. 62 mmol/g cell/h of glucose uptake rate, the simulated specific growth rate was 0. 089 h" , a little lower than the experimental value 0. 11 h " . In addition, the accuracy of the single gene deletion simulation in pyrimidine metabolism reached 90% . [ Conclusion] The final metabolic network model covered the biochemical information of citrate cycle, glycolysis, pentose phosphate pathway, fatty acid metabolism, vitamin Bl2 biosynthesis and amino acid biosynthesis, and reflected the effect of substrates and genes on the growth of the bacterium to a certain extent.%[目的]通过挖掘实验性文献,建立巨大芽胞杆菌事实型代谢网络模型,以详尽解析生理特性,优化其生理功能.[方法]从PubMed、Derwent Innovations Index、中国知网等公共文献(专利)数据库中获取与巨大芽胞杆菌(Bacillus megaterium)相关的实验性文献建立本地文献数据库.采用文献挖掘工具获取功能基因、酶、代谢物和生化反应

  15. 艰难梭状杆菌肠毒素B在巨大芽孢杆菌中的表达和鉴定%Expression and identification of recombinant Clostridium difficile toxin B using Bacillus megaterium system

    Institute of Scientific and Technical Information of China (English)

    杨桂林; 刘威龙; 姚红艳; 周伯平; 冯汉平

    2011-01-01

    目的 获得高纯度和具有生物活性的重组肠毒素B(rTcdB).方法 以艰难梭状芽孢杆菌染色体DNA为模板,通过PCR方法扩增得到全长TcdB基因并克隆至穿梭载体pHis1522.构建的质粒经直接测序验证无误后,转化巨大芽孢杆菌原生质体,在木糖的诱导作用下进行TcdB的表达、纯化及生物活性鉴定.结果 从细菌培养液中纯化得到rTcdB,浓度达到5~10 mg/L,其相对分子质量与天然的TcdB蛋白接近,生物活性与天然的TcdB蛋白相似.结论 在巨大芽孢杆菌中成功表达了具有完整结构和活性的艰难梭状芽孢杆菌TcdB重组蛋白.%Objective To express and purify recombinant and biologically active Clostridium difficile toxin B (rTcdB). Methods The genes of TcdB were amplified by polymerase chain reaction (PCR) using chromosomal DNA from a toxigenic strain, and cloned into a shuttle vector pHis1522.The sequences of TcdB genes in the vector were verified by DNA sequencing. The construction was transformed into Bacillus megaterium protoplasts and the protein expression was driven by a xylose promoter. The purified protein was tested for biological activity. Results rTcdB was successfully purified from bacterial crude extracts. Approximately 5-10 mg of highly purified recombinant toxin was obtained from one liter of bacterial culture. The expressed rTcdB had molecular mass similar to the native toxin, and its biological activity was proved to be similar to its native counterpart after an extensive examination. Conclusion rTcdB with biological activities is successfully expressed in Bacillus megaterium.

  16. 巨大芽孢杆菌MBFF6产絮凝剂培养条件的优化及絮凝剂的化学性质%Optimization of culture conditions for bioflocculant-producing Bacillus megaterium MBFF6 and its chemical characteristics

    Institute of Scientific and Technical Information of China (English)

    曹芳; 余秀梅; 郭娟利; 杨晓尧; 潘康成

    2012-01-01

    At present, flocculants are prevalent in the variety of industrial processes such as wastewater treatment, drinking water purification and downstream processing in fermentation processes. In general, flocculants are classified into three groups: inorganic flocculants, organic synthetic flocculants and naturally occurring flocculants. Bioflocculants have received considerable scientific and biotechnological attention in recent years because of their biodegradability, harmlessness and lack of secondary pollution of their degradation intermediates. Compared with inorganic flocculants and synthetic flocculants, bioflocculants have not yet been produced industrially because of their low flocculating capability and high cost. In order to reduce costs of production, research is needed to identify and select new bioflocculant-producing microorganisms and strains capable of utilizing low-cost substrates, and to learn how to optimize fermentation conditions to enhance bioflocculant activity. We had previously screened a bioflocculant-producing strain identified as Bacillus megaterium MBFF6, and the strain producing bioflocculant was named as BF6. In this study, Optimization for culture conditions of strain MBFF6 and chemical characteristics of bioflocculant BF6 were studied. The effect of culture time, carbon source, nitrogen source, C/N ratio, pH, metal ions, culture temperature and inoculum size on bioflocculant production by 5. megaterium MBFF6 was investigated by signal factor test. At the same time, thermal stability and chemical characteristics of bioflocculant were studied. The bioflocculant BF6 was purified from supernatant by ethanol precipitation. Whether containing α-amino acids and polysaccharide of the purified bioflocculant was analyzed by ninhydrin reaction and Molish and anthrone reaction. Proteins and nucleic acids were qualitatively determined by UV spectrophotometry. The chemical groups of purified bioflocculant were analyzed using a Fourier

  17. Effects of Bacillus megaterium 1259 Contrast with Yucca and Bacillus subtillis on Performance and Ammonia Nitrogen Content in Excrement of Laying Hens%巨大芽孢杆菌1259制剂对比丝兰素、枯草芽孢杆菌对产蛋鸡生产性能及排泄物中氨氮含量的影响

    Institute of Scientific and Technical Information of China (English)

    丁文骏; 王强; 戴美梅; 张为利; 张艳云

    2016-01-01

    本文旨在比较饲粮中添加巨大芽孢杆菌1259( BM1259)制剂与丝兰素、枯草芽孢杆菌对产蛋鸡生产性能及排泄物中氨氮含量的影响。选用384只300日龄健康、产蛋率相近的海兰褐蛋鸡,随机分为4组,每组6个重复,每个重复16只鸡。对照组饲喂基础饲粮,试验组分别在基础饲粮中添加100 mg/kg BM1259制剂、丝兰素和枯草芽孢杆菌。预试期14 d,正试期42 d。结果表明:1)与对照组相比,BM1259组极显著提高了总产蛋重、产蛋数、产蛋率和平均蛋重( P<0.01),极显著降低了料蛋比( P<0.01),其结果与枯草芽孢杆菌组相近,优于丝兰素组。2)与对照组相比,BM1259组极显著降低了蛋鸡排泄物中氨态氮、尿酸、尿素氮含量和脲酶活性( P<0.01),其结果与丝兰素组相近,优于枯草芽孢杆菌组。3)饲粮中添加BM1259制剂相对于添加丝兰素或枯草芽孢杆菌能够获得更大的经济效益。由此可见,在蛋鸡饲粮中添加BM1259制剂能提高生产性能、增加利润并具备较优的除氨除臭作用。%This research was carried out to investigate the effects of Bacillus megaterium 1259 ( BM1259) con⁃trast with Yucca and Bacillus subtillis on performance and the content of ammonia nitrogen in excrement of lay⁃ing hens. A total of three hundred and eighty four 300⁃day⁃old healthy Hy⁃line variety brown laying hens with similar laying rate were selected and randomly assigned into four groups with six replicates per group and six⁃teen hens per replicate. Birds in the control group were fed a basal diet, and the others in experimental groups were fed the basal diet supplemented with 100 mg/kg BM1259, Yucca and Bacillus subtillis, respectively. The preliminary period lasted for 14 days and the experiment lasted for 42 days. The results showed as follows:1) compared with the control group, the total egg mass, egg production, laying

  18. STUDY ON STRENGTH OF BACTERIAL CONCRETE IN BACILLUS MEGATERIUM

    OpenAIRE

    Deepika.B*, C.Savinth Kumar

    2016-01-01

    Concrete a strong, durable material composed of cement, aggregate and water is the most used building material in the world. Concrete has an ultimate load bearing capacity under compression but the material is weak in tension. The steel reinforced bars take the load when the concrete cracks in tension. To increase the strength and durability of the structure either crack that are formed should be repaired conventionally using epoxy injection or latex treatment or by providing extra reinforcem...

  19. Efficient transformation of Bacillus thuringiensis requires nonmethylated plasmid DNA.

    OpenAIRE

    Macaluso, A; Mettus, A M

    1991-01-01

    The transformation efficiency of Bacillus thuringiensis depends upon the source of plasmid DNA. DNA isolated from B. thuringiensis, Bacillus megaterium, or a Dam- Dcm- Escherichia coli strain efficiently transformed several B. thuringiensis strains, B. thuringiensis strains were grouped according to which B. thuringiensis backgrounds were suitable sources of DNA for transformation of other B. thuringiensis strains, suggesting that B. thuringiensis strains differ in DNA modification and restri...

  20. In vivo and in vitro digestibility of plant ingredients and diets by Bacillus phytases in tilapia, Oreochromis mossambicus

    Directory of Open Access Journals (Sweden)

    Rande B. Dechavez

    2012-12-01

    Full Text Available This study aimed to evaluate four Bacillus phytases for their efficacy in making plant-baseddiets bioavailable to tilapia (Oreochromis mossambicus using in vivo digestibility measurement and todetermine the in vitro level of dephosphorylation. The four Bacillus strains used were B. pumilus , B. megaterium , B. coagulans, and B. licheniformis. Phytase activities varied between bacterial sources aswell as between feed ingredients. For the cassava leaf meal, Pi released was highest in B. pumilus andwas not significantly different from those of B. megaterium and B. licheniformis. For the soybean meal, Pirelease was in this decreasing order: B. megaterium > B. pumilus > B. coagulans > B. licheniformisphytase. For the corn meal, addition of B. licheniformis phytase to the reaction mixture resulted insignificantly the highest Pi released followed by B. coagulans phytase which was not significantly differentfrom that of B. megaterium phytase which released the lowest Pi. Pi released by B. pumilus phytase fromcorn meal was not significantly different from the lowest Pi release of B. megaterium phytase. Theapparent digestibility coefficient (ADC values for the feed dry matter (DM ranged from 86.3 to 88.3%and were not significantly different from each other (p > 0.05.

  1. Enantioselective Hydrolysis of Phenyl Glycidyl Ether Catalyzed by Newly Isolated Bacillus Megaterium ECU1001

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@Microbial epoxide hydrolases from bacterial and fungal sources?1? are hi ghly versatile catalysts for the asymmetric hydrolysis of chiral epoxides which are extensively employed as useful building blocks for the synthesis of various biologically active products in the pharmaceutical and agrochemical industries. Microorganism means allows an unlimited supply of these enzymes for preparative -scale applications. Phenyl glycidyl ether (PGE), an aryl epoxide, is a potenti ally useful compound in the synthesis of chiral amino alcohols and bioactive com pounds such as ?blockers. No suitable biocatalyst with sufficiently high enan tioselectivity (E?20) for the kinetic resolution of this compound was previ ously found among bacteria and fungi. This prompted us to screen epoxide hydrola se-producing microorganisms with higher enantioselectivity toward phenyl glycid yl ether from soil samples.

  2. Genome Sequence of Bacillus endophyticus and Analysis of Its Companion Mechanism in the Ketogulonigenium vulgare-Bacillus Strain Consortium.

    Directory of Open Access Journals (Sweden)

    Nan Jia

    Full Text Available Bacillus strains have been widely used as the companion strain of Ketogulonigenium vulgare in the process of vitamin C fermentation. Different Bacillus strains generate different effects on the growth of K. vulgare and ultimately influence the productivity. First, we identified that Bacillus endophyticus Hbe603 was an appropriate strain to cooperate with K. vulgare and the product conversion rate exceeded 90% in industrial vitamin C fermentation. Here, we report the genome sequencing of the B. endophyticus Hbe603 industrial companion strain and speculate its possible advantage in the consortium. The circular chromosome of B. endophyticus Hbe603 has a size of 4.87 Mb with GC content of 36.64% and has the highest similarity with that of Bacillus megaterium among all the bacteria with complete genomes. By comparing the distribution of COGs with that of Bacillus thuringiensis, Bacillus cereus and B. megaterium, B. endophyticus has less genes related to cell envelope biogenesis and signal transduction mechanisms, and more genes related to carbohydrate transport and metabolism, energy production and conversion, as well as lipid transport and metabolism. Genome-based functional studies revealed the specific capability of B. endophyticus in sporulation, transcription regulation, environmental resistance, membrane transportation, extracellular proteins and nutrients synthesis, which would be beneficial for K. vulgare. In particular, B. endophyticus lacks the Rap-Phr signal cascade system and, in part, spore coat related proteins. In addition, it has specific pathways for vitamin B12 synthesis and sorbitol metabolism. The genome analysis of the industrial B. endophyticus will help us understand its cooperative mechanism in the K. vulgare-Bacillus strain consortium to improve the fermentation of vitamin C.

  3. Genome Sequence of Bacillus endophyticus and Analysis of Its Companion Mechanism in the Ketogulonigenium vulgare-Bacillus Strain Consortium.

    Science.gov (United States)

    Jia, Nan; Du, Jin; Ding, Ming-Zhu; Gao, Feng; Yuan, Ying-Jin

    2015-01-01

    Bacillus strains have been widely used as the companion strain of Ketogulonigenium vulgare in the process of vitamin C fermentation. Different Bacillus strains generate different effects on the growth of K. vulgare and ultimately influence the productivity. First, we identified that Bacillus endophyticus Hbe603 was an appropriate strain to cooperate with K. vulgare and the product conversion rate exceeded 90% in industrial vitamin C fermentation. Here, we report the genome sequencing of the B. endophyticus Hbe603 industrial companion strain and speculate its possible advantage in the consortium. The circular chromosome of B. endophyticus Hbe603 has a size of 4.87 Mb with GC content of 36.64% and has the highest similarity with that of Bacillus megaterium among all the bacteria with complete genomes. By comparing the distribution of COGs with that of Bacillus thuringiensis, Bacillus cereus and B. megaterium, B. endophyticus has less genes related to cell envelope biogenesis and signal transduction mechanisms, and more genes related to carbohydrate transport and metabolism, energy production and conversion, as well as lipid transport and metabolism. Genome-based functional studies revealed the specific capability of B. endophyticus in sporulation, transcription regulation, environmental resistance, membrane transportation, extracellular proteins and nutrients synthesis, which would be beneficial for K. vulgare. In particular, B. endophyticus lacks the Rap-Phr signal cascade system and, in part, spore coat related proteins. In addition, it has specific pathways for vitamin B12 synthesis and sorbitol metabolism. The genome analysis of the industrial B. endophyticus will help us understand its cooperative mechanism in the K. vulgare-Bacillus strain consortium to improve the fermentation of vitamin C.

  4. Investigation and Analysis of Bacillus in Yogurt Production Environment%酸奶生产环境中芽孢杆菌的调查与分析

    Institute of Scientific and Technical Information of China (English)

    康博燕; 牟光庆; 陈历俊; 姜铁民

    2013-01-01

    According to the studies on the microbial community of yogurt production environment,found that the Bacillus.sp was the majority bacteria.Base on physiological and biochemical identification and molecular identification on the bacillus isolate from sampling plots,found that Bacillus subtilis account for 30%,Bacillus licheniformis account for 19%,Bacillus megaterium account for 14%,The rest of the seven kinds of the bacillus account for 37%.By traceability analysis,found that there was cross contamination in the connected workshop.%通过对某厂酸奶生产环境微生物菌群分析,发现此环境中微生物以芽孢杆菌属(Bacillus.sp)的菌种居多.对各个采样点分离、纯化的芽孢杆菌进行生理生化和分子鉴定,结果为枯草芽孢杆菌(Bacillus subtilis)占所分离鉴定芽孢杆菌的30%,地衣芽孢杆菌(Bacillus licheniformis)占19%,巨大芽孢杆菌(Bacillus megaterium)占14%,其余7种菌共占37%.对它们进行溯源分析,发现有连通的车间存在交叉污染.

  5. [The flotation characteristics of Bacillus cells and spores].

    Science.gov (United States)

    Stabnikova, E V; Gregirchak, N N; Taranenko, T O

    1991-01-01

    Variations in hydrophobicity of the surface of bacillary cells and their capacity to flotation in the process of batch cultivation have been studied. It is shown that hydrophobicity of the cell surface increases in the course of batch cultivation of Bacillus thuringiensis, B. licheniformis and B. megaterium. Hydrophobicity of spores of the mentioned cultures is considerably higher than that of the vegetative cells. The increase of hydrophobicity of bacillary cells positively correlated with their capacity to flotation. That is why the use of flotation for the age fractionation of bacillary cells is possible: spores are concentrated in the foam while vegetative cells remain in the culture liquid. PMID:1779906

  6. Fungicin M4: a narrow spectrum peptide antibiotic from Bacillus licheniformis M-4.

    Science.gov (United States)

    Lebbadi, M; Gálvez, A; Maqueda, M; Martínez-Bueno, M; Valdivia, E

    1994-07-01

    The strain Bacillus licheniformis M-4 produces a 3.4 kDa hydrophilic peptide with antifungal activity, named fungicin M4. Analysis of the purified peptide shows that it contains the amino acids Glu (8), Arg (5), Pro (4), Tyr (8), Val (3), Met (2) and Orn (4). Its inhibitory spectrum is restricted to Microsporum canis CECT 2797, Mucor mucedo CECT 2653, Mucor plumbeus CCM 443, Sporothrix schenckii CECT 2799, Bacillus megaterium and Corynebacterium glutamicum CECT 78. Fungicin M4 exerts biocidal activity on liquid cultures of Sporothrix schenckii CECT 2799.

  7. Isolation and physico-chemical characterization of an antifungal and antibacterial peptide produced by Bacillus licheniformis A12.

    Science.gov (United States)

    Gálvez, A; Maqueda, M; Martínez-Bueno, M; Lebbadi, M; Valdivia, E

    1993-07-01

    An antifungal substance named peptide A12-C has been purified to homogeneity from supernatants of sporulated cultures of Bacillus licheniformis A12. It consists of a 0.77-kDa hydrophilic peptide containing two residues of Glu and one of Arg, Ala, Pro, Tyr and Orn. No fatty acids, phosphorus or carbohydrates have been detected. Peptide A12-C is active on several fungi (Microsporum canis CECT 2797, Mucor mucedo CECT 2653, M. plumbeus (CCM F 443, Sporothrix schenckii CECT 2799 and Trichophyton mentagrophytes CECT 2793) and bacteria (Bacillus megaterium, Corynebacterium glutamicum, Sarcina and Mycobacterium), although the latter are less sensitive.

  8. Physiology of Sporolactobacillus strains isolated from different habitats and the indication of in vitro antagonism against Bacillus species.

    Science.gov (United States)

    Holzapfel, W H; Botha, S J

    1988-10-01

    In an ecological study only low numbers of Sporolactobacillus were found in habitats such as the faeces of herbivores, the rumen of cattle and the final waste water of an abattoir. Their presence in the final waste water of an abattoir indicates their possible association with food, and, more specifically, with meat. Differences were found in some physiological characteristics. One isolate (L2404) differed from the authentic Sporolactobacillus ATCC 15538 by its inability to ferment inulin, its growth in presence of 6.5% NaCl and in 0.2% tellurite, by the isomer(s) of lactic acid produced and the mol% G + G in the DNA. One Sporolactobacillus isolate (L2407) showed antagonism against Bacillus cereus, Bacillus cereus var, mycoides, Bacillus megaterium and Bacillus subtilis. PMID:3275317

  9. Effect of Bacillus spp. on seed germination of selected species of the genus Cuscuta (Convolvulaceae

    Directory of Open Access Journals (Sweden)

    Fatemeh Hadizadeh

    2014-04-01

    Full Text Available Species of the genus Cuscuta are annual angiospermic rootless and leafless (achlorophyllous parasitic plants. Bacillus is an example of PGPR bacteria exhibiting plant growth promoting activity. In this study the effects of bacterial suspension on germination of dodder’s seed has been determinated. Seeds of three Cuscuta species were collected from field for evaluating effects of three different Bacillus on its germination. Results show that seed germination of the C. monogyna and C. campestris is inhibited by all three bacterial species. Based on Tukey analysis, the highest inhibitory activity on seed germination of C. monogyna was shown with B. pumilus (68.88%; as well as C. campestris with B. megaterium (95.76% and B. pumilus (91.53%, whilst seed germination of C. europaea was almost identically inhibited by all three bacterial species. This paper reports the variable effects of Bacillus species on the seed germination of selected Cuscuta species.

  10. Production and characterization of phytase from Bacillus spp. as feed additive in aquaculture

    Directory of Open Access Journals (Sweden)

    Rande B. Dechavez

    2011-07-01

    Full Text Available Phytases are phosphohydrolases that catalyze the release of phosphate from phytate (myo inositol hexakisphosphate, the major phosphorus (P form mostly occurring in animal feeds of plant origin. These enzymes can be supplemented in animal diets to reduce inorganic phosphorus supplementation and fecal phosphorus excretion. Four species of Bacillus namely, B. pumilus , B.megaterium , B. coagulans , and B. licheniformis were used to study the biochemical characteristics of their phytases. All the strains investigated were able to hydrolyze extracellular phytate. The activity of phytase increased markedly at the late stationary phase in all the species tested. Highest enzyme activity was found in phytase from B. megaterium after the 4th day of culture. The crude phytases from the different Bacillus strains were optimally active at pH values ranging 5.5 to 7.0 at 37 0 C and retained their activity at temperatures up to 80 0 C. The enzymes exhibited thermostability, retaining ~50 %activity at 70 0 C and were fairly stable up to pH 10. These properties indicate that the Bacillus phytases appear to be suitable for animal feed supplementation in aquaculture to improve the bioavailability of phosphorus.

  11. Phylogeny of marine Bacillus isolates from the Gulf of Mexico

    Science.gov (United States)

    Siefert, J. L.; Larios-Sanz, M.; Nakamura, L. K.; Slepecky, R. A.; Paul, J. H.; Moore, E. R.; Fox, G. E.; Jurtshuk, P. Jr

    2000-01-01

    The phylogeny of 11 pigmented, aerobic, spore-forming isolates from marine sources was studied. Forty-two biochemical characteristics were examined, and a 16S rDNA sequence was obtained for each isolate. In a phylogenetic tree based on 16S sequencing, four isolates (NRRL B-14850, NRRL B-14904, NRRL B-14907, and NRRL B-14908) clustered with B. subtilis and related organisms; NRRL B-14907 was closely related to B. amyloliquefaciens. NRRL B-14907 and NRRL B-14908 were phenotypically similar to B. amyloliquefaciens and B. pumilus, respectively. Three strains (NRRL B-14906, NRRL B-14910, and NRRL B-14911) clustered in a clade that included B. firmus, B. lentus, and B. megaterium. NRRL B-14910 was closely related phenotypically and phylogenetically to B. megaterium. NRRL B-14905 clustered with the mesophilic round spore-producing species, B. fusiformis and B. sphaericus; the isolate was more closely related to B. fusiformis. NRRL B-14905 displayed characteristics typical of the B. sphaericus-like organisms. NRRL B-14909 and NRRL B-14912 clustered with the Paenibacillus species and displayed characteristics typical of the genus. Only NRRL B-14851, an unusually thin rod that forms very small spores, may represent a new Bacillus species.

  12. Bacillus anthracis

    OpenAIRE

    2003-01-01

    The events of 11 September 2001 and the subsequent anthrax outbreaks have shown that the West needs to be prepared for an increasing number of terrorist attacks, which may include the use of biological warfare. Bacillus anthracis has long been considered a potential biological warfare agent, and this review will discuss the history of its use as such. It will also cover the biology of this organism and the clinical features of the three disease forms that it can produce: cutaneous, gastrointe...

  13. Bacillus anthracis

    OpenAIRE

    BOSERET, GÉRALDINE; Linden, Annick; Mainil, Jacques

    2002-01-01

    The literature describes several methods for detection of Bacillus anthracis based on application of specific bacteriophages. The following methods of pahoinpitely are used to identify the causative agent of anthrax: the reaction of bacteriophage titer growth (RBTG), the reaction of phage adsorption (RPA), fagoterapii method (FTM) and fluorescentserological method (FSM). The essence of RBTG consists in the following: if there is the researchform of bacteria presents in the test material, then...

  14. Rapid inactivation of seven Bacillus spp. under simulated Mars UV irradiation

    Science.gov (United States)

    Schuerger, Andrew C.; Richards, Jeff T.; Newcombe, David A.; Venkateswaran, Kasthuri

    2006-03-01

    Seven Bacillus spp. were exposed to simulations of Mars-normal UV fluence rates in order to study the effects of UV irradiation on microbial survival. A UV illumination system was calibrated to deliver 9.78 W m -2 (35.2 kJ m -2 h -1) of UVC + UVB irradiation (200-320 nm) to microbial samples, thus creating a clear-sky simulation (0.5 optical depth) of equatorial Mars. The Bacillus spp. studied were: B. licheniformis KL-196, B. megaterium KL-197, B. nealsonii FO-092, B. pumilus FO-36B, B. pumilus SAFR-032, B. subtilis 42HS1, and B. subtilis HA101. The bacteria were prepared as thin monolayers of endospores on aluminum coupons in order to simulate contaminated spacecraft surfaces. Bacterial monolayers were exposed to Mars UV irradiation for time-steps of 0, 0.25, 0.5, 1, 5, 15, 30, 60, 120, or 180 min. The surviving endospores were then assayed with a Most Probable Numbers (MPN) procedure and with a culture-based assay that utilized a bacillus spore germination medium. Results indicated that B. pumilus SAFR-032 was the most resistant, and B. subtilis 42HS-1 and B. megaterium were the most sensitive of the seven strains exposed to martian UV fluence rates. Bacillus subtilis 42HS1 and B. megaterium were inactivated after 30 min exposure to Mars UV, while B. pumilus SAFR-032 required 180 min for full inactivation in both assays. Spores of B. pumilus SAFR-032 exhibited significantly different inactivation kinetics suggesting that this wild type isolate also was more resistant than the standard dosimetric strain, B. subtilis HA101. Although the various Bacillus spp. exhibited diverse levels of UV resistance, none were immune to UV irradiation, and, thus, all species would be expected to be inactivated on Sun-exposed spacecraft surfaces within a few tens-of-minutes to a few hours on sol 1 under clear-sky conditions on equatorial Mars. The inactivation kinetics of all seven Bacillus spp. support the conclusion that significant levels of bioload reductions are possible on

  15. Phylogeny in aid of the present and novel microbial lineages: diversity in Bacillus.

    Directory of Open Access Journals (Sweden)

    Shalini Porwal

    Full Text Available Bacillus represents microbes of high economic, medical and biodefense importance. Bacillus strain identification based on 16S rRNA sequence analyses is invariably limited to species level. Secondly, certain discrepancies exist in the segregation of Bacillus subtilis strains. In the RDP/NCBI databases, out of a total of 2611 individual 16S rDNA sequences belonging to the 175 different species of the genus Bacillus, only 1586 have been identified up to species level. 16S rRNA sequences of Bacillus anthracis (153 strains, B. cereus (211 strains, B. thuringiensis (108 strains, B. subtilis (271 strains, B. licheniformis (131 strains, B. pumilus (83 strains, B. megaterium (47 strains, B. sphaericus (42 strains, B. clausii (39 strains and B. halodurans (36 strains were considered for generating species-specific framework and probes as tools for their rapid identification. Phylogenetic segregation of 1121, 16S rDNA sequences of 10 different Bacillus species in to 89 clusters enabled us to develop a phylogenetic frame work of 34 representative sequences. Using this phylogenetic framework, 305 out of 1025, 16S rDNA sequences presently classified as Bacillus sp. could be identified up to species level. This identification was supported by 20 to 30 nucleotides long signature sequences and in silico restriction enzyme analysis specific to the 10 Bacillus species. This integrated approach resulted in identifying around 30% of Bacillus sp. up to species level and revealed that B. subtilis strains can be segregated into two phylogenetically distinct groups, such that one of them may be renamed.

  16. Measurements of the Ultraviolet Fluorescence Cross Sections and Spectra of Bacillus Anthracis Simulants

    Energy Technology Data Exchange (ETDEWEB)

    Stephens, J.R.

    1998-09-01

    Measurements of the ultraviolet autofluorescence spectra and absolute cross sections of the Bacillus anthracis (Ba) simulants Bacillus globigii (Bg), Bacillus megaterium (Bm), Bacillus subtilis (Bs), and Bacillus cereus (Bc) were measured. Fluorescence spectra and cross sections of pine pollen (Pina echinata) were measured for comparison. Both dried vegetative cells and spores separated from the sporulated vegetative material were studied. The spectra were obtained by suspending a small number (<10) of particles in air in our Single Particle Spectroscopy Apparatus (SPSA), illuminating the particles with light from a spectrally filtered arc lamp, and measuring the fluorescence spectra of the particles. The illumination was 280 nm (20 nm FWHM) and the fluorescence spectra was measured between 300 and 450 nm. The fluorescence cross section of vegetative Bg peaks at 320 nm with a maximum cross section of 5 X 10{sup -14} cm{sup 2}/sr-nm-particle while the Bg spore fluorescence peaks at 310 nm with peak fluorescence of 8 X 10{sup -15} cm{sup 2}/sr-nm-particle. Pine pollen particles showed a higher fluorescence peaking at 355 nm with a cross section of 1.7 X 10{sup -13} cm{sup 2}/sr-nm-particle. Integrated cross sections ranged from 3.0 X 10{sup -13} for the Bg spores through 2.25 X 10{sup -12} (cm{sup 2}/sr-particle) for the vegetative cells.

  17. Cellular fatty acid profile and H(+)-ATPase activity to assess acid tolerance of Bacillus sp. for potential probiotic functional attributes.

    Science.gov (United States)

    Shobharani, P; Halami, Prakash M

    2014-11-01

    The present study has been focused widely on comparative account of probiotic qualities of Bacillus spp. for safer usage. Initially, 170 heat resistant flora were isolated and selected for non-pathogenic cultures devoid of cytK, hblD, and nhe1 virulence genes. Subsequently, through biochemical tests along with 16S rRNA gene sequencing and fatty acid profiling, the cultures were identified as Bacillus megaterium (AR-S4), Bacillus subtilis (HR-S1), Bacillus licheniformis (Csm1-1a and HN-S1), and Bacillus flexus (CDM4-3c and CDM3-1). The selected cultures showed 70-80 % survival under simulated gastrointestinal condition which was also confirmed through H(+)-ATPase production. The amount of H(+)-ATPase increased by more than 2-fold when grown at pH 2 which support for the acid tolerance ability of Bacillus isolates. The study also examined the influence of acidic pH on cellular fatty acid composition of Bacillus spp. A remarkable shift in the fatty acid profile was observed at acidic pH through an increased amount of even numbered fatty acid (C16 and C18) in comparison with odd numbered (C15 and C17). Additionally, the cultures exhibited various probiotic functional properties. Overall, the study increases our understanding of Bacillus spp. and will allow both industries and consumers to choose for well-defined probiotic with possible health benefits. PMID:25125040

  18. A method for the determination of bacterial spore DNA content based on isotopic labelling, spore germination and diphenylamine assay; ploidy of spores of several Bacillus species

    International Nuclear Information System (INIS)

    A reliable method for measuring the spore DNA content, based on radioactive DNA labelling, spore germination in absence of DNA replication and diphenylamine assay, was developed. The accuracy of the method, within 10 - 15%, is adequate for determining the number of chromosomes per spore, provided that the genome size is known. B subtilis spores were shown to be invariably monogenomic, while those of larger bacilli Bacillus megaterium, Bacillus cereus and Bacillus thuringiensis, often, if not invariably, contain two genomes. Attempts to modify the spore DNA content of B subtilis by altering the richness of the sporulation medium, the sporulation conditions (liquid or solid medium), or by mutation, were apparently unsuccessful. An increase of spore size with medium richness, not accompanied by an increase in DNA content, was observed. The implication of the apparently species-specific spore ploidy and the influence of the sporulation conditions on spore size and shape are discussed

  19. Diversity of bacteria of the genus Bacillus on board of international space station.

    Science.gov (United States)

    Alekhova, T A; Zakharchuk, L M; Tatarinova, N Yu; Kadnikov, V V; Mardanov, A V; Ravin, N V; Skryabin, K G

    2015-01-01

    From swabs of surfaces of equipment and air samples of the Russian segment of the International Space Station, nine strains of spore-forming bacteria of the genus Bacillus belonging to the species B. pumilus, B. licheniformis, B. subtilis, B. megaterium, and B. amyloliquefaciens were isolated. The last species of bacilli on the equipment of RS ISS was detected for the first time. For these species of bacilli, there are known strains that can be opportunistic to humans, and their metabolites can cause biodegradation of equipment and materials. B. pumilus found on ISS belongs to the group of bacteria that exhibits a particularly high resistance to adverse environmental conditions, such as dehydration, ultraviolet and gamma radiation, and chemical disinfection.

  20. Diversity of bacteria of the genus Bacillus on board of international space station.

    Science.gov (United States)

    Alekhova, T A; Zakharchuk, L M; Tatarinova, N Yu; Kadnikov, V V; Mardanov, A V; Ravin, N V; Skryabin, K G

    2015-01-01

    From swabs of surfaces of equipment and air samples of the Russian segment of the International Space Station, nine strains of spore-forming bacteria of the genus Bacillus belonging to the species B. pumilus, B. licheniformis, B. subtilis, B. megaterium, and B. amyloliquefaciens were isolated. The last species of bacilli on the equipment of RS ISS was detected for the first time. For these species of bacilli, there are known strains that can be opportunistic to humans, and their metabolites can cause biodegradation of equipment and materials. B. pumilus found on ISS belongs to the group of bacteria that exhibits a particularly high resistance to adverse environmental conditions, such as dehydration, ultraviolet and gamma radiation, and chemical disinfection. PMID:26728721

  1. BACILLUS MEGATERIUM 501RIF INTRODUCED INTO THE SOIL: FACTORS AFFECTING THE RATE OF SURVIVAL, SPORULATION AND DECOMPOSITION OF THE HERBICIDE PROMETRYN

    OpenAIRE

    YU, V; T.O. LISINA

    2014-01-01

    Introduction of microorganisms into the soil is a fundamental problem in application of microbial preparations in agriculture, crop production and ecology. Its effectiveness depends on many factors (i.e. the type and strain of microorganism, the physical and chemical properties of the soil, plant cover, climate and so on), which are not well understood, leading to a high variability of the results of application of microbial preparations. In this article, there is reported the first investiga...

  2. Inactivation of Spores of Bacillus anthracis Sterne, Bacillus cereus, and Bacillus thuringiensis subsp. israelensis by Chlorination

    OpenAIRE

    Rice, E W; Adcock, N. J.; Sivaganesan, M; Rose, L. J.

    2005-01-01

    Three species of Bacillus were evaluated as potential surrogates for Bacillus anthracis for determining the sporicidal activity of chlorination as commonly used in drinking water treatment. Spores of Bacillus thuringiensis subsp. israelensis were found to be an appropriate surrogate for spores of B. anthracis for use in chlorine inactivation studies.

  3. Isolation, screening and identifi cation of Bacillus spp. as direct-fed microbial candidates for aflatoxin B1 biodegradation

    Institute of Scientific and Technical Information of China (English)

    Rosario; Galarza-Seeber; Juan; David; Latorre; Xochitl; Hernandez-Velasco; Amanda; Drake; Wolfenden; Lisa; Renee; Bielke; Anita; Menconi; Billy; Marshall; Hargis; Guillermo; Tellez

    2015-01-01

    Objective: To evaluate the ability of Bacillus spp. as direct-fed microbials(DFM) to biodegrade al atoxin B1(AFB1) by using an in vitro digestive model simulating in vivo conditions.Methods: Sixty-nine Bacillus isolates were obtained from intestines, and soil samples were screened by using a selective media method against 0.25 and 1.00 μg/m L of AFB1 in modii ed Czapek-Dox medium. Plates were incubated at 37 °C and observed every two days for two weeks. Physiological properties of the three Bacillus spp. candidates were characterized biochemically and by 16 S r RNA sequence analyzes for identii cation. Tolerance to acidic p H, osmotic concentrations of Na Cl, bile salts were tested, and antimicrobial sensitivity proi les were also determined. Bacillus candidates were individually sporulated by using a solid fermentation method and combined. Spores were incorporated into 1 of 3 experimental feed groups: 1) Negative control group, with unmedicated starter broiler feed without AFB1; 2) Positive control group, with negative control feed contaminated with 0.01% AFB1; 3) DFM treated group, with positive control feed supplemented with 109 spores/g. After digestion time(3:15 h), supernatants and digesta were collected for high-performance liquid chromatography l uorescence detection analysis by triplicate.Results: Three out of those sixty-nine DFM candidates showed ability to biodegrade AFB1 in vitro based on growth as well as reduction of l uorescence and area of clearance around each colony in modii ed Czapek-Dox medium which was clearly visible under day light after 48 h of evaluation. Analysis of 16S-DNA identii ed the strains as Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus subtilis. The three Bacillus strains were tolerant to acidic conditions(p H 2.0), tolerant to a high osmotic pressure(Na Cl at 6.5%), and were able to tolerate 0.037% bile salts after 24 h of incubation. No signii cant dif erences(P > 0.05) were observed in the concentrations of

  4. Isolation, screening and identification of Bacillus spp. as direct-fed microbial candidates for aflatoxin B1 biodegradation

    Institute of Scientific and Technical Information of China (English)

    Rosario Galarza-Seeber; Juan David Latorre; Xochitl Hernandez-Velasco; Amanda Drake Wolfenden; Lisa Renee Bielke; Anita Menconi; Billy Marshall Hargis; Guillermo Tellez

    2015-01-01

    To evaluate the ability of Bacillus spp. as direct-fed microbials (DFM) to biodegrade aflatoxin B1 (AFB1) by using an in vitro digestive model simulating in vivo conditions. Methods: Sixty-nine Bacillus isolates were obtained from intestines, and soil samples were screened by using a selective media method against 0.25 and 1.00 µg/mL of AFB1 in modified Czapek-Dox medium. Plates were incubated at 37 °C and observed every two days for two weeks. Physiological properties of the three Bacillus spp. candidates were characterized biochemically and by 16S rRNA sequence analyzes for identification. Tolerance to acidic pH, osmotic concentrations of NaCl, bile salts were tested, and antimicrobial sensitivity profiles were also determined. Bacillus candidates were individually sporulated by using a solid fermentation method and combined. Spores were incorporated into 1 of 3 experimental feed groups: 1) Negative control group, with unmedicated starter broiler feed without AFB1; 2) Positive control group, with negative control feed contaminated with 0.01% AFB1; 3) DFM treated group, with positive control feed supplemented with 109 spores/g. After digestion time (3:15 h), supernatants and digesta were collected for high-performance liquid chromatography fluorescence detection analysis by triplicate. Results: Three out of those sixty-nine DFM candidates showed ability to biodegrade AFB1 in vitro based on growth as well as reduction of fluorescence and area of clearance around each colony in modified Czapek-Dox medium which was clearly visible under day light after 48 h of evaluation. Analysis of 16S-DNA identified the strains as Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus subtilis. The three Bacillus strains were tolerant to acidic conditions (pH 2.0), tolerant to a high osmotic pressure (NaCl at 6.5%), and were able to tolerate 0.037%bile salts after 24 h of incubation. No significant differences (P > 0.05) were observed in the concentrations of AFB1 in

  5. Comparison of Bacillus thuringiensis and Bacillus cereus

    International Nuclear Information System (INIS)

    Bacillus cereus and Bacillus thuringiensis are closely related, spore forming soil bacteria. B. thuringiensis produces insecticidal crystal proteins during sporulation and these toxins are the most important biopesticides in the world today. Genomes of the B. thuringiensis and B. cereus strains were analysed by pulsed field gel electrophoresis after treatment of the DNA with the restriction enzyme NotI. The NotI fingerprint patterns varied both within the B. thuringiensis and the B. cereus strains. The size of the fragments varied between 15 and 1350 kb. When physical maps of the B. thuringiensis and B. cereus strains were compared, B. thuringiensis appeared to be as closely related to B. cereus as the B. cereus strains were to each other. Nine out of 12 B. thuringiensis strains and 18 out of 25 B. cereus strains produced enterotoxins. The close relationship between B. thuringiensis and B. cereus should be taken into consideration when B. thuringiensis is used as a biopesticide. (author). 10 refs, 4 figs, 1 tab

  6. Characterization of Bacillus cereus

    NARCIS (Netherlands)

    Wijnands LM; Dufrenne JB; Leusden FM; MGB

    2002-01-01

    Bacillus cereus is a ubiquitary microorganism that may cause food borne disease. Pathogenicity, however, depends on various characteristics such as the ability to form (entero)-toxin(s) that can not be detected by microbiological methods. Further characterization of pathogenic properties is not only

  7. Biodiversity in Bacillus cereus

    NARCIS (Netherlands)

    Pielaat A; Fricker M; Nauta MJ; Leusden FM van; MGB

    2006-01-01

    Experiments have been performed by different partners to identify variability in properties of Bacillus cereus strains that contribute to the extent of their virulence as part of an EU project. To this end, 100 B. cereus strains were selected and screened for biological properties, such as toxin pro

  8. Essential Bacillus subtilis genes

    DEFF Research Database (Denmark)

    Kobayashi, K.; Ehrlich, S.D.; Albertini, A.;

    2003-01-01

    To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. Among approximate to4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. Another 79 genes we...

  9. Taxonomy Icon Data: Bacillus subtilis [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available g Bacillus_subtilis_S.png Bacillus_subtilis_NS.png http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Bacillus...+subtilis&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Bacillus+subtilis&t=NL http://biosciencedbc.jp/taxonomy..._icon/icon.cgi?i=Bacillus+subtilis&t=S http://biosciencedbc.jp/taxonomy..._icon/icon.cgi?i=Bacillus+subtilis&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=214 ...

  10. Identification and Properties of Amylase-producing Bacillus Isolated from Songhe Distiller's Yeast%宋河酒曲中产淀粉酶芽孢菌的分离鉴定及产酶特性

    Institute of Scientific and Technical Information of China (English)

    侯小歌; 王俊英; 张杰; 李学思; 李绍亮; 胡炳义

    2012-01-01

    为了有效控制宋河酒曲的制曲过程和发酵进程,对宋河酒曲中产淀粉酶芽孢菌进行了分离鉴定并研究筛选菌株的产淀粉酶特性.结果表明:从宋河酒曲中共分离到12株产淀粉酶芽孢菌,均属革兰氏阳性杆状菌,初步鉴定归为1个属5个种,即芽孢杆菌属(Bacillus)、坚硬芽孢杆菌(Bacillus firmus)、枯草芽孢杆菌( Bacillus subtilis)、地衣芽孢杆菌(Bacillus licheni formis)、凝结芽孢杆菌(Bacillus coagulans)、巨大芽孢杆菌(Bacillus megaterium),其中,坚硬芽孢杆菌是宋河大曲产淀粉酶芽孢菌的数量优势菌群,获得1株地衣芽孢杆菌SQ2为中温型高产淀粉酶菌株,其液态发酵产酶特性为37℃培养,前24 h产酶较弱,此后,酶活力迅速上升,72 h达到最大值为89 μg/(mL·min),产酶旺盛期发生在菌体成熟期和衰亡初期,产酶过程pH值先稍偏酸性后接近中性.%Amylase-producing bacilli isolated from Songhe Distiller's yeast were identified and their properties were studied to effectively control the starter-making process and fermentation process. The results showed that 12 bacillus strains which could produce amylase were isolated and identified as Bacillus firmus, Bacillus subtilis, Bacillus licheniformis, Bacillus coagulans and Bacillus megaterium , which belonged to genus of Bacillus. The B. firmus was the dominant amylase-producing bacillus in Songhe Daqu. Strain SQ2 was obtained as a high producing-amylase bacillus by determining transparent circle, its properties of producing amylase brothing under 37℃ were showed as follows: amylase-producing activity kept minimum in first fermenting 24 hours, gradually raised after 24 hours, reached primary stabilizing value to 72 hours, and decreased gently after fermenting 72 hours, the maximum value of amylase activity could reach 89 μg/(mL · min), the maximum amylase production occurred at the stage of strain maturity and decline growth. The pH value kept slightly acid

  11. Fluorescent Amplified Fragment Length Polymorphism Analysis of Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis Isolates

    OpenAIRE

    Hill, Karen K.; Ticknor, Lawrence O.; Okinaka, Richard T.; Asay, Michelle; Blair, Heather; Bliss, Katherine A.; Laker, Mariam; Pardington, Paige E.; Richardson, Amber P.; Tonks, Melinda; Beecher, Douglas J.; Kemp, John D.; Kolstø, Anne-Brit; Wong, Amy C. Lee; Keim, Paul

    2004-01-01

    DNA from over 300 Bacillus thuringiensis, Bacillus cereus, and Bacillus anthracis isolates was analyzed by fluorescent amplified fragment length polymorphism (AFLP). B. thuringiensis and B. cereus isolates were from diverse sources and locations, including soil, clinical isolates and food products causing diarrheal and emetic outbreaks, and type strains from the American Type Culture Collection, and over 200 B. thuringiensis isolates representing 36 serovars or subspecies were from the U.S. D...

  12. Genome analysis shows Bacillus axarquiensis is not a later heterotypic synonym of Bacillus mojavensis; reclassification of Bacillus malacitensis and Brevibacterium halotolerans as heterotypic synonyms of Bacillus axarquiensis.

    Science.gov (United States)

    Dunlap, Christopher A; Bowman, Michael J; Schisler, David A; Rooney, Alejandro P

    2016-06-01

    Bacillus axarquiensis and Bacillus malacitensis were previously reported to be later heterotypic synonyms of Bacillus mojavensis, based primarily on DNA-DNA relatedness values. We have sequenced draft genomes of Bacillus axarquiensis NRRL B-41617T and Bacillus malacitensis NRRL B-41618T. Comparative genomics and DNA-DNA relatedness calculations showed that while Bacillus axarquiensis and Bacillus malacitensis are synonymous with each other, they are not synonymous with Bacillus mojavensis. In addition, a draft genome was completed for Brevibacterium halotolerans, a strain long suspected of being a Bacillus subtilis group member based on 16S rRNA similarities (99.8 % with Bacillus mojavensis). Comparative genomics and DNA-DNA relatedness calculations showed that Brevibacterium halotolerans is synonymous with Bacillus axarquiensis and Bacillus malacitensis. The pairwise in silico DNA-DNA hybridization values calculated in comparisons between the three conspecific strains were all greater than 92 %, which is well above the standard species threshold of 70 %. While the pairwise in silico DNA-DNA hybridization values calculated in comparisons of the three conspecific strains with Bacillus mojavensis were all less than 65 %. The combined results of our genotype and phenotype studies showed that Bacillus axarquiensis, Bacillus malacitensis and Brevibacterium halotolerans are conspecific and distinct from Bacillus mojavensis. Because the valid publication of the name Bacillus axarquiensis predates the publication of the name Bacillus malacitensis, we propose that Bacillus malacitensis be reclassified as a synonym of Bacillus axarquiensis. In addition, we propose to reclassify Brevibacterium halotolerans as a synonym of Bacillus axarquiensis. An amended description of Bacillus axarquiensis is provided.

  13. Genome analysis shows Bacillus axarquiensis is not a later heterotypic synonym of Bacillus mojavensis; reclassification of Bacillus malacitensis and Brevibacterium halotolerans as heterotypic synonyms of Bacillus axarquiensis.

    Science.gov (United States)

    Dunlap, Christopher A; Bowman, Michael J; Schisler, David A; Rooney, Alejandro P

    2016-06-01

    Bacillus axarquiensis and Bacillus malacitensis were previously reported to be later heterotypic synonyms of Bacillus mojavensis, based primarily on DNA-DNA relatedness values. We have sequenced draft genomes of Bacillus axarquiensis NRRL B-41617T and Bacillus malacitensis NRRL B-41618T. Comparative genomics and DNA-DNA relatedness calculations showed that while Bacillus axarquiensis and Bacillus malacitensis are synonymous with each other, they are not synonymous with Bacillus mojavensis. In addition, a draft genome was completed for Brevibacterium halotolerans, a strain long suspected of being a Bacillus subtilis group member based on 16S rRNA similarities (99.8 % with Bacillus mojavensis). Comparative genomics and DNA-DNA relatedness calculations showed that Brevibacterium halotolerans is synonymous with Bacillus axarquiensis and Bacillus malacitensis. The pairwise in silico DNA-DNA hybridization values calculated in comparisons between the three conspecific strains were all greater than 92 %, which is well above the standard species threshold of 70 %. While the pairwise in silico DNA-DNA hybridization values calculated in comparisons of the three conspecific strains with Bacillus mojavensis were all less than 65 %. The combined results of our genotype and phenotype studies showed that Bacillus axarquiensis, Bacillus malacitensis and Brevibacterium halotolerans are conspecific and distinct from Bacillus mojavensis. Because the valid publication of the name Bacillus axarquiensis predates the publication of the name Bacillus malacitensis, we propose that Bacillus malacitensis be reclassified as a synonym of Bacillus axarquiensis. In addition, we propose to reclassify Brevibacterium halotolerans as a synonym of Bacillus axarquiensis. An amended description of Bacillus axarquiensis is provided. PMID:27030978

  14. Bacillus thuringiensis (Bt)

    Science.gov (United States)

    2004-01-01

    Bacillus thuringiensis (Bt), a natural bacteria found all over the Earth, has a fairly novel way of getting rid of unwanted insects. Bt forms a protein substance (shown on the right) that is not harmful to humans, birds, fish or other vertebrates. When eaten by insect larvae the protein causes a fatal loss of appetite. For over 25 years agricultural chemical companies have relied heavily upon safe Bt pesticides. New space based research promises to give the insecticide a new dimension in effectiveness and applicability. Researchers from the Consortium for Materials Development in Space along with industrial affiliates such as Abott Labs and Pern State University flew Bt on a Space Shuttle mission in the fall of 1996. Researchers expect that the Shuttle's microgravity environment will reveal new information about the protein that will make it more effective against a wider variety of pests.

  15. Production of Cellulases, Xylanase, Pectinase, alpha-amylase and Protease Enzymes Cocktail by Bacillus spp. and Their Mixed Cultures with Candida tropicalis and Rhodotorula glutinis under Solid State Fermentation

    International Nuclear Information System (INIS)

    A group of twelve locally isolated Bacillus species, B.megaterium (MAI and MA II), B.licheniformis (MLI and ML II); B. circulans, B. stearothermophilis, B.cereus, B.sphaericus, B. pumilus, B. laterosporus, B. coagulans and B. pantothenticus, were examined for the production of cellulases, xylanase, pectinase, alpha-amylase and protease enzymes cocktail on wheat bran under solid state fermentation (SSF). All species were found to be potent hydrolyzing enzymes producers and the superior producing species were B. megaterium MAI and B. licheniformis. On the other hand, both of them still produced highest enzyme titres when mixed with Candida tropicalis or Rhodotorula glutinis, yeast strains. The two superior bacterial strains produced the highest enzymatic activities when coculturing with C. tropicalis compared with coculturing with R. glutinis only or with both C. tropicalis and R. glutinis in combination. The inferior activities of cocultures (B. megaterinm MAI and R. glutinis) were enhanced in carboxymethyl cellulase (CMCase), filter paper cellulase (FPase), avecilase, xylanase, pectinase, -amylase and protease by gamma irradiation at dose 1.0 kGy with percent increase 8 %, 20 %, 10 %, 4 %, 31 %, 22 % and 34 %, respectively as compared with un-irradiated cocultures

  16. Genome Differences That Distinguish Bacillus anthracis from Bacillus cereus and Bacillus thuringiensis

    OpenAIRE

    Radnedge, Lyndsay; Agron, Peter G.; Hill, Karen K.; Jackson, Paul J.; Ticknor, Lawrence O; Keim, Paul; Andersen, Gary L.

    2003-01-01

    The three species of the group 1 bacilli, Bacillus anthracis, B. cereus, and B. thuringiensis, are genetically very closely related. All inhabit soil habitats but exhibit different phenotypes. B. anthracis is the causative agent of anthrax and is phylogenetically monomorphic, while B. cereus and B. thuringiensis are genetically more diverse. An amplified fragment length polymorphism analysis described here demonstrates genetic diversity among a collection of non-anthrax-causing Bacillus speci...

  17. N-terminal amino acid sequence of Bacillus licheniformis alpha-amylase: comparison with Bacillus amyloliquefaciens and Bacillus subtilis Enzymes.

    OpenAIRE

    Kuhn, H; Fietzek, P P; Lampen, J. O.

    1982-01-01

    The thermostable, liquefying alpha-amylase from Bacillus licheniformis was immunologically cross-reactive with the thermolabile, liquefying alpha-amylase from Bacillus amyloliquefaciens. Their N-terminal amino acid sequences showed extensive homology with each other, but not with the saccharifying alpha-amylases of Bacillus subtilis.

  18. Functional Feed Assessment on Litopenaeus vannamei Using 100% Fish Meal Replacement by Soybean Meal, High Levels of Complex Carbohydrates and Bacillus Probiotic Strains

    Directory of Open Access Journals (Sweden)

    Rosalia Contreras

    2011-06-01

    Full Text Available Functional feed supplemented with alternative-economic nutrient sources (protein, carbohydrates, lipids and probiotics are being considered in shrimp/fish aquaculture production systems as an option to increase yield and profits and to reduce water pollution. In this study the probiotic potential to formulate functional feeds have been evaluated using four dietary treatments: Treatment 1 (B + Bs; Bacillus subtilis potential probiotic strain was supplemented to a soybeanmeal (SBM—carbohydrates (CHO basal feed. Treatment 2 (B + Bm; Bacillus megaterium potential probiotic strain was supplemented to the same SBM-CHO basal feed. In Treatment 3 (B; SBM-CHO basal feed was not supplemented with probiotic strains. Treatment 4 (C; fishmeal commercial feed (FM was utilized as positive control. Feeding trials evaluated the survival, growth, and food conversion ratio and stress tolerance of juvenile Litopenaeus vannamei (Boone Pacific white shrimp. Best overall shrimp performance was observed for animals fed with Treatment 1 (B+Bs; additionally, stress tolerance and hemolymph metabolites also showed the best performance in this treatment. SBM-CHO basal feed not supplemented with probiotic strains (B presented smaller growth and lower feed conversion ratio (FCR. Shrimps fed with the fishmeal commercial feed (C presented the lowest stress tolerance to high ammonia and low oxygen levels. Specifically selected B. subtilis strains are recommended to formulate functional and economical feeds containing high levels of vegetable; protein and carbohydrates as main dietary sources in L. vannamei cultures.

  19. Isolation of Mexican Bacillus Species and Their Effects in Promoting Growth of Chili Pepper (Capsicum annuum L. cv Jalapeño).

    Science.gov (United States)

    Peña-Yam, Laura P; Ruíz-Sánchez, Esaú; Barboza-Corona, José E; Reyes-Ramírez, Arturo

    2016-09-01

    The purpose of this work was to isolate and identify native bacteria from plants collected in the State of Yucatán, México with the ability to promote growth of chili pepper (Capsicum annuum L. cv Jalapeño). We identified nine bacterial isolates that belong to five species of Bacillus (i.e. Bacillus subtilis, B. flexus, B. cereus, B. megaterium and B. endophyticus) that produced indoleacetic acid (4.0-24.3 µg/mL) with solubilization index of 1.3-1.6. All the bacterial isolates were evaluated based on their ability to promote growth of chili pepper. Plants inoculated with B. subtilis ITC-N67 showed an increase in stem diameter and root volume, whereas inoculation with B. cereus ITC-BL18 increased the number of flower buds, fresh biomass of roots and total fresh biomass. Conversely, B. flexus ITC-P4 and B. flexus ITC-P22 showed deleterious effect on root volume and total biomass. In summary, our data showed that native B. cereus TC-BL18 and B. subtilis ITC-N67 have potential to be used as growth promoting microorganism for chili pepper, particularly in the state of Yucatán, México. PMID:27407303

  20. Phages Preying on Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis: Past, Present and Future

    Directory of Open Access Journals (Sweden)

    Annika Gillis

    2014-07-01

    Full Text Available Many bacteriophages (phages have been widely studied due to their major role in virulence evolution of bacterial pathogens. However, less attention has been paid to phages preying on bacteria from the Bacillus cereus group and their contribution to the bacterial genetic pool has been disregarded. Therefore, this review brings together the main information for the B. cereus group phages, from their discovery to their modern biotechnological applications. A special focus is given to phages infecting Bacillus anthracis, B. cereus and Bacillus thuringiensis. These phages belong to the Myoviridae, Siphoviridae, Podoviridae and Tectiviridae families. For the sake of clarity, several phage categories have been made according to significant characteristics such as lifestyles and lysogenic states. The main categories comprise the transducing phages, phages with a chromosomal or plasmidial prophage state, γ-like phages and jumbo-phages. The current genomic characterization of some of these phages is also addressed throughout this work and some promising applications are discussed here.

  1. Phages preying on Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis: past, present and future.

    Science.gov (United States)

    Gillis, Annika; Mahillon, Jacques

    2014-07-01

    Many bacteriophages (phages) have been widely studied due to their major role in virulence evolution of bacterial pathogens. However, less attention has been paid to phages preying on bacteria from the Bacillus cereus group and their contribution to the bacterial genetic pool has been disregarded. Therefore, this review brings together the main information for the B. cereus group phages, from their discovery to their modern biotechnological applications. A special focus is given to phages infecting Bacillus anthracis, B. cereus and Bacillus thuringiensis. These phages belong to the Myoviridae, Siphoviridae, Podoviridae and Tectiviridae families. For the sake of clarity, several phage categories have been made according to significant characteristics such as lifestyles and lysogenic states. The main categories comprise the transducing phages, phages with a chromosomal or plasmidial prophage state, γ-like phages and jumbo-phages. The current genomic characterization of some of these phages is also addressed throughout this work and some promising applications are discussed here. PMID:25010767

  2. Environmental Persistence of Bacillus anthracis and Bacillus subtilis Spores

    Science.gov (United States)

    Wood, Joseph P.; Meyer, Kathryn M.; Kelly, Thomas J.; Choi, Young W.; Rogers, James V.; Riggs, Karen B.; Willenberg, Zachary J.

    2015-01-01

    There is a lack of data for how the viability of biological agents may degrade over time in different environments. In this study, experiments were conducted to determine the persistence of Bacillus anthracis and Bacillus subtilis spores on outdoor materials with and without exposure to simulated sunlight, using ultraviolet (UV)-A/B radiation. Spores were inoculated onto glass, wood, concrete, and topsoil and recovered after periods of 2, 14, 28, and 56 days. Recovery and inactivation kinetics for the two species were assessed for each surface material and UV exposure condition. Results suggest that with exposure to UV, decay of spore viability for both Bacillus species occurs in two phases, with an initial rapid decay, followed by a slower inactivation period. The exception was with topsoil, in which there was minimal loss of spore viability in soil over 56 days, with or without UV exposure. The greatest loss in viable spore recovery occurred on glass with UV exposure, with nearly a four log10 reduction after just two days. In most cases, B. subtilis had a slower rate of decay than B. anthracis, although less B. subtilis was recovered initially. PMID:26372011

  3. FORMALDEHYDE GAS INACTIVATION OF BACILLUS ANTHRACIS, BACILLUS SUBTILIS AND GEOBACILLUS STEAROTHERMOPHILUS SPORES ON INDOOR SURFACE MATERIALS.

    Science.gov (United States)

    Research evaluated the decontamination of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface material using formaldehyde gas. Spores were dried on seven types of indoor surfaces and exposed to 1100 ppm formaldehyde gas for 10 hr. Fo...

  4. Screening of Bacillus Species with Potentials of Antibiotics Production

    Directory of Open Access Journals (Sweden)

    Faruk Adamu KUTA

    2009-07-01

    Full Text Available Sixteen soil samples were collected from different refuse dump sites in Minna, the capital Niger State, and analysed for the presence of Bacillus species. Physical-chemical analysis of the soil samples revealed the followings: PH value 6.89-8.47; moisture content 1.58 – 21.21% and temperature 27-28ºC. Using both pour plate and streak method of inoculation, total bacterial count in the soil samples ranged from 3.8×104 cfu/g 16.0×104 cfu/g. The identified Bacillus species included: Bacillus cereus (30.8%, Bacillus brevis (1.9% Bacillus polymyxa (3.8%, Bacillus lichenifomis (13.5%, Bacillus spherericus (7.7%, Bacillus mycoides (13.5%, Bacillus pumilus (7.7%, Bacillus subtilis (3.8%, Bacillus alvei (1.9%, Bacillus laterosporous (1.9%, Bacillus firmus (9.6% and Bacillus circulars (3.8%. Antibiotic production tests indicated that nine Bacillus species out of twelve isolated in this study could be used to produce antibiotics that had effect on the test organisms. However, Bacillus polymyxa, Bacillus sphaericus and Bacillus laterosporous had little or no effect on the tested organisms. This study suggests that some Bacillus species have potential to produce high quality antibiotics that can be use to control microbial growth in future.

  5. Cyclic Lipopeptide Biosynthetic Genes and Products, and Inhibitory Activity of Plant-Associated Bacillus against Phytopathogenic Bacteria.

    Directory of Open Access Journals (Sweden)

    Isabel Mora

    Full Text Available The antibacterial activity against bacterial plant pathogens and its relationships with the presence of the cyclic lipopeptide (cLP biosynthetic genes ituC (iturin, bmyB (bacillomycin, fenD (fengycin and srfAA (surfactin, and their corresponding antimicrobial peptide products have been studied in a collection of 64 strains of Bacillus spp. isolated from plant environments. The most frequent antimicrobial peptide (AMP genes were bmyB, srfAA and fenD (34-50% of isolates. Most isolates (98.4% produced surfactin isoforms, 90.6% iturins and 79.7% fengycins. The antibacterial activity was very frequent and generally intense among the collection of strains because 75% of the isolates were active against at least 6 of the 8 bacterial plant pathogens tested. Hierarchical and correspondence analysis confirmed the presence of two clearly differentiated groups. One group consisted of Bacillus strains that showed a strong antibacterial activity, presented several cLPs genes and produced several isoforms of cLPs simultaneously, mainly composed of B. subtilis and B. amyloliquefaciens, although the last one was exclusive to this group. Another group was characterized by strains with very low or none antibacterial activity, that showed one or none of the cLP genes and produced a few or none of the corresponding cLPs, and was the most heterogenous group including B. subtilis, B. licheniformis, B. megaterium, B. pumilus, B. cereus and B. thuringiensis, although the last two were exclusive to this group. This work demonstrated that the antagonistic capacity of plant-associated Bacillus against plant pathogenic bacteria is related to the presence of cLP genes and to the production of the corresponding cLPs, and it is mainly associated to the species B. subtilis and B. amyloliquefaciens. Our findings would help to increase the yield and efficiency of screening methods to obtain candidate strains to biocontrol agents with a mechanism of action relaying on the

  6. Cyclic Lipopeptide Biosynthetic Genes and Products, and Inhibitory Activity of Plant-Associated Bacillus against Phytopathogenic Bacteria.

    Science.gov (United States)

    Mora, Isabel; Cabrefiga, Jordi; Montesinos, Emilio

    2015-01-01

    The antibacterial activity against bacterial plant pathogens and its relationships with the presence of the cyclic lipopeptide (cLP) biosynthetic genes ituC (iturin), bmyB (bacillomycin), fenD (fengycin) and srfAA (surfactin), and their corresponding antimicrobial peptide products have been studied in a collection of 64 strains of Bacillus spp. isolated from plant environments. The most frequent antimicrobial peptide (AMP) genes were bmyB, srfAA and fenD (34-50% of isolates). Most isolates (98.4%) produced surfactin isoforms, 90.6% iturins and 79.7% fengycins. The antibacterial activity was very frequent and generally intense among the collection of strains because 75% of the isolates were active against at least 6 of the 8 bacterial plant pathogens tested. Hierarchical and correspondence analysis confirmed the presence of two clearly differentiated groups. One group consisted of Bacillus strains that showed a strong antibacterial activity, presented several cLPs genes and produced several isoforms of cLPs simultaneously, mainly composed of B. subtilis and B. amyloliquefaciens, although the last one was exclusive to this group. Another group was characterized by strains with very low or none antibacterial activity, that showed one or none of the cLP genes and produced a few or none of the corresponding cLPs, and was the most heterogenous group including B. subtilis, B. licheniformis, B. megaterium, B. pumilus, B. cereus and B. thuringiensis, although the last two were exclusive to this group. This work demonstrated that the antagonistic capacity of plant-associated Bacillus against plant pathogenic bacteria is related to the presence of cLP genes and to the production of the corresponding cLPs, and it is mainly associated to the species B. subtilis and B. amyloliquefaciens. Our findings would help to increase the yield and efficiency of screening methods to obtain candidate strains to biocontrol agents with a mechanism of action relaying on the production of

  7. Genome-Based Genetic Tool Development for Bacillus methanolicus: Theta- and Rolling Circle-Replicating Plasmids for Inducible Gene Expression and Application to Methanol-Based Cadaverine Production

    Science.gov (United States)

    Irla, Marta; Heggeset, Tonje M. B.; Nærdal, Ingemar; Paul, Lidia; Haugen, Tone; Le, Simone B.; Brautaset, Trygve; Wendisch, Volker F.

    2016-01-01

    Bacillus methanolicus is a thermophilic methylotroph able to overproduce amino acids from methanol, a substrate not used for human or animal nutrition. Based on our previous RNA-seq analysis a mannitol inducible promoter and a putative mannitol activator gene mtlR were identified. The mannitol inducible promoter was applied for controlled gene expression using fluorescent reporter proteins and a flow cytometry analysis, and improved by changing the -35 promoter region and by co-expression of the mtlR regulator gene. For independent complementary gene expression control, the heterologous xylose-inducible system from B. megaterium was employed and a two-plasmid gene expression system was developed. Four different replicons for expression vectors were compared with respect to their copy number and stability. As an application example, methanol-based production of cadaverine was shown to be improved from 11.3 to 17.5 g/L when a heterologous lysine decarboxylase gene cadA was expressed from a theta-replicating rather than a rolling-circle replicating vector. The current work on inducible promoter systems and compatible theta- or rolling circle-replicating vectors is an important extension of the poorly developed B. methanolicus genetic toolbox, valuable for genetic engineering and further exploration of this bacterium.

  8. Triple fixation of Bacillus subtilis dormant spores.

    OpenAIRE

    Kozuka, S; Tochikubo, K

    1983-01-01

    A triple-fixation method with a sequential application of 5% glutaraldehyde, 1% osmium tetroxide, and 2% potassium permanganate gave superior preservation of the ultrastructure of Bacillus subtilis dormant spores with a thick spore coat.

  9. Narrow terahertz attenuation signatures in Bacillus thuringiensis.

    Science.gov (United States)

    Zhang, Weidong; Brown, Elliott R; Viveros, Leamon; Burris, Kellie P; Stewart, C Neal

    2014-10-01

    Terahertz absorption signatures from culture-cultivated Bacillus thuringiensis were measured with a THz photomixing spectrometer operating from 400 to 1200 GHz. We observe two distinct signatures centered at ∼955 and 1015 GHz, and attribute them to the optically coupled particle vibrational resonance (surface phonon-polariton) of Bacillus spores. This demonstrates the potential of the THz attenuation signatures as "fingerprints" for label-free biomolecular detection. PMID:23821459

  10. Narrow terahertz attenuation signatures in Bacillus thuringiensis.

    Science.gov (United States)

    Zhang, Weidong; Brown, Elliott R; Viveros, Leamon; Burris, Kellie P; Stewart, C Neal

    2014-10-01

    Terahertz absorption signatures from culture-cultivated Bacillus thuringiensis were measured with a THz photomixing spectrometer operating from 400 to 1200 GHz. We observe two distinct signatures centered at ∼955 and 1015 GHz, and attribute them to the optically coupled particle vibrational resonance (surface phonon-polariton) of Bacillus spores. This demonstrates the potential of the THz attenuation signatures as "fingerprints" for label-free biomolecular detection.

  11. Hydrazine vapor inactivates Bacillus spores

    Science.gov (United States)

    Schubert, Wayne W.; Engler, Diane L.; Beaudet, Robert A.

    2016-05-01

    NASA policy restricts the total number of bacterial spores that can remain on a spacecraft traveling to any planetary body which might harbor life or have evidence of past life. Hydrazine, N2H4, is commonly used as a propellant on spacecraft. Hydrazine as a liquid is known to inactivate bacterial spores. We have now verified that hydrazine vapor also inactivates bacterial spores. After Bacillus atrophaeus ATCC 9372 spores deposited on stainless steel coupons were exposed to saturated hydrazine vapor in closed containers, the spores were recovered from the coupons, serially diluted, pour plated and the surviving bacterial colonies were counted. The exposure times required to reduce the spore population by a factor of ten, known as the D-value, were 4.70 ± 0.50 h at 25 °C and 2.85 ± 0.13 h at 35 °C. These inactivation rates are short enough to ensure that the bioburden of the surfaces and volumes would be negligible after prolonged exposure to hydrazine vapor. Thus, all the propellant tubing and internal tank surfaces exposed to hydrazine vapor do not contribute to the total spore count.

  12. Production of amylolytic enzymes by bacillus spp

    International Nuclear Information System (INIS)

    Sixty six bacteria and twenty fungi were isolated from various sources. These varied from rotten fruites to local drinks and soil samples from different parts of Sudan. On the basis of index of amylolytic activity, forty one bacteria and twelve fungi were found to hydrolyse strach. The best ten strach hydrolysing isolates were identified all as bacilli (Bacillus licheniformis SUD-K1, SUD-K2, SUD-K4, SUD-O, SUD-SRW, SUD-BRW, SUD-By, Bacillus subtilis SUD-K3, and Bacillus circulans SUD-D and SUD-K7). Their amylase productivity was studied with respect to temperature and time. Amylolytic activity was measured by spectrophotometer, the highest activity was produced in around 24 hours of growth in all; six of which gave the highest amylase activity at 50 deg C and the rest at 45C. Based on the thermal production six isolates were chosen for further investigation. These were Bacillus licheniformis SUD-K1, SUD-K2, SUD-K4, SUD-O, Bacillus subtilis SUD-K3 and Bacillus circulans SUD-K7. The inclusion of strach and Mg++ ions in the culture medium gave the highest enzyme yield. The Ph 9.0 was found to be the optimum for amylase production for all isolates except Bacillus subtilis SUD-K3 which had an optimum at pH 7.0. Three isolates (Bacillus licheniformis SUD-K1, SUD-K4 and SUD-O recorded highestamylase production in a medium supplemented with peptone while the rest (Bacillus licheniformis SUD-K2, Bacillus subtilis SUD-K3 and Bacillus circulans SUD-K7) gave highest amylase productivity in a medium supplemented with malt extract. Four isolates (Bacillus licheniformis SUD-K1 and Bacillus subtilis SUD-K3 gave maximum amylase production in a medium containing 0.5% soluble strach while the rest (gave maximum amylase production at 2%. Soluble strach was found to be best substrate among the different carbon sources tested. The maximum temperature for amylase activity ranged from 60-70 deg C and 1% strach concentration was optimum for all isolates. Addition of different metal ions

  13. DECONTAMINATION ASSESSMENT OF BACILLUS ANTHRACIS, BACILLUS SUBTILIS, AND GEOBACILLUS STEAROTHERMOPHILUS SPORES ON INDOOR SURFACTS USING A HYDROGEN PERIOXIDE GAS GENERATOR

    Science.gov (United States)

    Aims: To evaluate the decontamination of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface materials using hydrogen peroxide gas. Methods and Results: B. anthracis, B. subtilis, and G. Stearothermophilus spores were dried on seven...

  14. Bacillus cereus as a nongastrointestinal pathogen

    Directory of Open Access Journals (Sweden)

    Pavani G.

    2014-02-01

    Full Text Available The potential of Bacillus cereus to cause systemic infections is of serious concern. Apart from Gastrointestinal infections, it causes respiratory tract infections, nosocomial infections, eye infections, CNS infections, cutaneous infections, endocarditis, osteomyelitis and urinary tract infections. The potential of this bacterium to cause life threatening infections has increased. Trauma is an important predisposing factor for Bacillus cereus infections. The maintenance of skin and mucous membrane integrity limits infection by this micro-organism. [Int J Res Med Sci 2014; 2(1.000: 28-30

  15. Comparative analysis of two-component signal transduction systems of Bacillus cereus, Bacillus thuringiensis and Bacillus anthracis

    NARCIS (Netherlands)

    Been, M.W.H.J. de; Francke, C.; Moezelaar, R.; Abee, T.; Siezen, R.J.

    2006-01-01

    Members of the Bacillus cereus group are ubiquitously present in the environment and can adapt to a wide range of environmental fluctuations. In bacteria, these adaptive responses are generally mediated by two-component signal transduction systems (TCSs), which consist of a histidine kinase (HK) and

  16. BOOK REVIEW – BACILLUS THURINGIENSIS: A CORNERSTONE OF MODERN AGRICULTURE BACILLUS THURINGIENSIS

    Science.gov (United States)

    Are you interested in the technical issues surrounding the use of Bacillus thuringiensis pesticidal traits as sprays and as plant incorporated protectants (transgenic crops)? Should the dimensions of human health, ecology, entomology, risk assessment, resistance management, and d...

  17. Differentiation of Bacillus anthracis from Bacillus cereus by gas chromatographic whole-cell fatty acid analysis.

    OpenAIRE

    Lawrence, D; Heitefuss, S; Seifert, H S

    1991-01-01

    Three strains of Bacillus anthracis and seven strains of Bacillus cereus were grown on complex medium and on synthetic medium. Gas chromatographic analysis of whole-cell fatty acids of strains grown on complex medium gave nearly identical fatty acid patterns. Fatty acid patterns of strains grown on synthetic medium showed a high content of branched-chain fatty acids. Significant differences between the fatty acid patterns of the two species were found. Odd iso/anteiso fatty acid ratios were a...

  18. Comparative genome analysis of Bacillus cereus group genomes withBacillus subtilis

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Iain; Sorokin, Alexei; Kapatral, Vinayak; Reznik, Gary; Bhattacharya, Anamitra; Mikhailova, Natalia; Burd, Henry; Joukov, Victor; Kaznadzey, Denis; Walunas, Theresa; D' Souza, Mark; Larsen, Niels; Pusch,Gordon; Liolios, Konstantinos; Grechkin, Yuri; Lapidus, Alla; Goltsman,Eugene; Chu, Lien; Fonstein, Michael; Ehrlich, S. Dusko; Overbeek, Ross; Kyrpides, Nikos; Ivanova, Natalia

    2005-09-14

    Genome features of the Bacillus cereus group genomes (representative strains of Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis sub spp israelensis) were analyzed and compared with the Bacillus subtilis genome. A core set of 1,381 protein families among the four Bacillus genomes, with an additional set of 933 families common to the B. cereus group, was identified. Differences in signal transduction pathways, membrane transporters, cell surface structures, cell wall, and S-layer proteins suggesting differences in their phenotype were identified. The B. cereus group has signal transduction systems including a tyrosine kinase related to two-component system histidine kinases from B. subtilis. A model for regulation of the stress responsive sigma factor sigmaB in the B. cereus group different from the well studied regulation in B. subtilis has been proposed. Despite a high degree of chromosomal synteny among these genomes, significant differences in cell wall and spore coat proteins that contribute to the survival and adaptation in specific hosts has been identified.

  19. Draft genome sequence of Bacillus endophyticus 2102.

    Science.gov (United States)

    Lee, Yong-Jik; Lee, Sang-Jae; Kim, Sun Hong; Lee, Sang Jun; Kim, Byoung-Chan; Lee, Han-Seung; Jeong, Haeyoung; Lee, Dong-Woo

    2012-10-01

    Bacillus endophyticus 2102 is an endospore-forming, plant growth-promoting rhizobacterium isolated from a hypersaline pond in South Korea. Here we present the draft sequence of B. endophyticus 2102, which is of interest because of its potential use in the industrial production of algaecides and bioplastics and for the treatment of industrial textile effluents. PMID:23012284

  20. Diagnostic Oligonucleotide Microarray Fingerprinting of Bacillus Isolates

    OpenAIRE

    Chandler, Darrell P.; Alferov, Oleg; Chernov, Boris; Daly, Don S; Golova, Julia; Perov, Alexander; Protic, Miroslava; Robison, Richard; Schipma, Matthew; White, Amanda; Willse, Alan

    2006-01-01

    A genome-independent microarray and new statistical techniques were used to genotype Bacillus strains and quantitatively compare DNA fingerprints with the known taxonomy of the genus. A synthetic DNA standard was used to understand process level variability and lead to recommended standard operating procedures for microbial forensics and clinical diagnostics.

  1. Complete Genome of Bacillus thuringiensis Myophage Spock

    OpenAIRE

    Maroun, Justin W.; Whitcher, Kelvin J.; Chamakura, Karthik R.; Kuty Everett, Gabriel F.

    2013-01-01

    Bacillus thuringiensis is a Gram-positive, sporulating soil microbe with valuable pesticide-producing properties. The study of bacteriophages of B. thuringiensis could provide new biotechnological tools for the use of this bacterium. Here, we present the complete annotated genome of Spock, a myophage of B. thuringiensis, and describe its features.

  2. Surfactin production by strains of Bacillus mojavensis

    Science.gov (United States)

    Bacillus mojavensis, RRC101 is an endophytic bacterium patented for control of fungal diseases in maize and other plants. DNA fingerprint analysis of the rep-PCR fragments of 35 B. mojavensis and 4 B. subtilis strains using the Diversilab genotyping system revealed genotypic distinctive strains alon...

  3. Protein-Tyrosine Phosphorylation in Bacillus subtilis

    DEFF Research Database (Denmark)

    Mijakovic, Ivan; Petranovic, Dina; Bottini, N.;

    2005-01-01

    phosphorylation, indicating that this post-translational modifi cation could regulate physiological processes ranging from stress response and exopolysaccharide synthesis to DNA metabolism. Some interesting work in this fi eld was done in Bacillus subtilis , and we here present the current state of knowledge...

  4. Phylogenomic analysis shows that ‘Bacillus vanillea’ is a later heterotypic synonym of Bacillus siamensis.

    Science.gov (United States)

    Dunlap, Christopher A

    2015-10-01

    Bacillus vanillea’ XY18 ( = CGMCC 8629 = NCCB 100507) was isolated from cured vanilla beans and involved in the formation of vanilla aroma compounds. A draft genome of this strain was assembled and yielded a length of 3.71 Mbp with a DNA G+C content of 46.3 mol%. Comparative genomic analysis with its nearest relatives showed only minor differences between this strain and the genome of the Bacillus siamensis KCTC 13613T ( = BCC 22614T = KACC 16244T), with a calculated DNA–DNA hybridization (DDH) value of 91.2 % and an average nucleotide identity (ANI) of 98.9 %. This DDH value is well above the recommended 70 % threshold for species delineation, as well as the ANI threshold of 95 %. In addition, the results of morphological, physiological, chemotaxonomic and phylogenetic analyses indicate that the type strains of these two taxa are highly similar with phenotype coherence. A core genome multi-locus sequencing analysis was conducted for the strains and the results show that ‘Bacillus vanillea’ XY18 clusters closely to the type strain of Bacillus siamensis. Therefore, it is proposed that the species ‘Bacillus vanillea’ XY18 ( = CGMCC 8629 = NCCB 100507) should be reclassified as a later heterotypic synonym of Bacillus siamensis KCTC 13613T ( = BCC 22614T = KACC 16244T). An emended description of Bacillus siamensis is provided. PMID:26296875

  5. Investigation of biosurfactant production by Bacillus pumilus 1529 and Bacillus subtilis WPI

    Directory of Open Access Journals (Sweden)

    shila khajavi shojaei

    2016-06-01

    Full Text Available Introduction: Biosurfactants are unique amphipathic molecules with extensive application in removing organic and metal contaminants. The purpose of this study was to investigate production of biosurfactant and determine optimal conditions to produce biosurfactant by Bacillus pumilus 1529 and Bacillus subtilis WPI. Materials and methods: In this study, effect of carbon source, temperature and incubation time on biosurfactant production was evaluated. Hemolytic activity, emulsification activity, oil spreading, drop collapse, cell hydrophobicity and measurement of surface tension were used to detect biosurfactant production. Then, according to the results, the optimal conditions for biosurfactant production by and Bacillus subtilis WPI was determined. Results: In this study, both bacteria were able to produce biosurfactant at an acceptable level. Glucose, kerosene, sugarcane molasses and phenanthrene used as a sole carbon source and energy for the mentioned bacteria. Bacillus subtilis WPI produced maximum biosurfactant in the medium containing kerosene and reduced surface tension of the medium to 33.1 mN/m after 156 hours of the cultivation at 37°C. Also, the highest surface tension reduction by Bacillus pumilus 1529 occurred in the medium containing sugarcane molasses and reduce the surface tension of culture medium after 156 hours at 37°C from 50.4 to 28.83 mN/m. Discussion and conclusion: Bacillus pumilus 1529 and Bacillus subtilis WPI had high potential in production of biosurfactant and degradation of petroleum hydrocarbons and Phenanthrene. Therefore, it could be said that these bacteria had a great potential for applications in bioremediation and other environmental process.

  6. Resistance to antimicrobials and acid and bile tolerance of Bacillus spp isolated from Bikalga, fermented seeds of Hibiscus sabdariffa

    DEFF Research Database (Denmark)

    Compaore, Clarisse S.; Jensen, Lars Bogø; Diawara, Brehima;

    2013-01-01

    In the aim of selecting starter cultures, thirteen species of Bacillus spp. including six Bacillus subtilis ssp. subtilis, four Bacillus licheniformis and three Bacillus amyloliquefaciens ssp. plantarum isolated from traditional Bikalga were investigated. The study included, for all isolates, gen...

  7. Bacillus caldolyticus prs gene encoding phosphoribosyldiphosphate synthase

    DEFF Research Database (Denmark)

    Krath, Britta N.; Hove-Jensen, Bjarne

    1996-01-01

    The prs gene, encoding phosphoribosyl-diphosphate (PRPP) synthase, as well as the flanking DNA sequences were cloned and sequenced from the Gram-positive thermophile, Bacillus caldolyticus. Comparison with the homologous sequences from the mesophile, Bacillus subtilis, revealed a gene (gca......D) encoding N-acetylglucosamine-l-phosphate uridyltransferase upstream of prs, and a gene homologous to ctc downstream of prs. cDNA synthesis with a B. caldolyticus gcaD-prs-ctc-specified mRNA as template, followed by amplification utilising the polymerase chain reaction indicated that the three genes are co......-transcribed. Comparison of amino acid sequences revealed a high similarity among PRPP synthases across a wide phylogenetic range. An E. coli strain harbouring the B. caldolyticus prs gene in a multicopy plasmid produced PRPP synthase activity 33-fold over the activity of a haploid B. caldolyticus strain. B. caldolyticus...

  8. Computational based functional analysis of Bacillus phytases.

    Science.gov (United States)

    Verma, Anukriti; Singh, Vinay Kumar; Gaur, Smriti

    2016-02-01

    Phytase is an enzyme which catalyzes the total hydrolysis of phytate to less phosphorylated myo-inositol derivatives and inorganic phosphate and digests the undigestable phytate part present in seeds and grains and therefore provides digestible phosphorus, calcium and other mineral nutrients. Phytases are frequently added to the feed of monogastric animals so that bioavailability of phytic acid-bound phosphate increases, ultimately enhancing the nutritional value of diets. The Bacillus phytase is very suitable to be used in animal feed because of its optimum pH with excellent thermal stability. Present study is aimed to perform an in silico comparative characterization and functional analysis of phytases from Bacillus amyloliquefaciens to explore physico-chemical properties using various bio-computational tools. All proteins are acidic and thermostable and can be used as suitable candidates in the feed industry. PMID:26672917

  9. Bacillus cereus Biofilms—Same, Only Different

    Science.gov (United States)

    Majed, Racha; Faille, Christine; Kallassy, Mireille; Gohar, Michel

    2016-01-01

    Bacillus cereus displays a high diversity of lifestyles and ecological niches and include beneficial as well as pathogenic strains. These strains are widespread in the environment, are found on inert as well as on living surfaces and contaminate persistently the production lines of the food industry. Biofilms are suspected to play a key role in this ubiquitous distribution and in this persistency. Indeed, B. cereus produces a variety of biofilms which differ in their architecture and mechanism of formation, possibly reflecting an adaptation to various environments. Depending on the strain, B. cereus has the ability to grow as immersed or floating biofilms, and to secrete within the biofilm a vast array of metabolites, surfactants, bacteriocins, enzymes, and toxins, all compounds susceptible to act on the biofilm itself and/or on its environment. Within the biofilm, B. cereus exists in different physiological states and is able to generate highly resistant and adhesive spores, which themselves will increase the resistance of the bacterium to antimicrobials or to cleaning procedures. Current researches show that, despite similarities with the regulation processes and effector molecules involved in the initiation and maturation of the extensively studied Bacillus subtilis biofilm, important differences exists between the two species. The present review summarizes the up to date knowledge on biofilms produced by B. cereus and by two closely related pathogens, Bacillus thuringiensis and Bacillus anthracis. Economic issues caused by B. cereus biofilms and management strategies implemented to control these biofilms are included in this review, which also discuss the ecological and functional roles of biofilms in the lifecycle of these bacterial species and explore future developments in this important research area. PMID:27458448

  10. Anthrose Biosynthetic Operon of Bacillus anthracis▿

    OpenAIRE

    Dong, Shengli; McPherson, Sylvia A.; Tan, Li; Chesnokova, Olga N.; Turnbough, Charles L.; Pritchard, David G.

    2008-01-01

    The exosporium of Bacillus anthracis spores consists of a basal layer and an external hair-like nap. The nap is composed primarily of the glycoprotein BclA, which contains a collagen-like region with multiple copies of a pentasaccharide side chain. This oligosaccharide possesses an unusual terminal sugar called anthrose, followed by three rhamnose residues and a protein-bound N-acetylgalactosamine. Based on the structure of anthrose, we proposed an enzymatic pathway for its biosynthesis. Exam...

  11. Antimicrobial Effects of Honey on Bacillus Cereus

    OpenAIRE

    This paper should be cited as: Javadzadeh M, Najafi M, Rezaei M, Dastoor M, Behzadi AS, Amiri A . [ Antimicrobial Effects of Honey on Bacillus Cereus ]. MLJ. 201 4 ; 8 ( 2 ): 55 - 61 [Article in Persian] Javadzadeh, M. (MSc; M Najafi; Rezaei, M. (MSc; Dastoor, M. (BSc; Behzadi, AS. (MSc; Amiri, A. (MSc

    2014-01-01

    Background and Objective: Honey is a healthy and nutritious food that has been used for a long time as a treatment for different diseases. One of the applied properties of honey is its antimicrobial effect, which differs between different types of honey due to variation of phenolic and antioxidant compositions. This study aimed to assess antimicrobial effect of honey on Bacillus cereus, considering its chemical properties. Material and Methods: Three samples of honey (A1 and A2 of Khorasan Ra...

  12. BACILLUS CEREUS: ISOLATION IN JENNET MILK

    Directory of Open Access Journals (Sweden)

    M.L. Scatassa

    2011-01-01

    Full Text Available Jennet milk as human food is hypoallergenic for patients affected by Cow Milk Protein Allergy and multiple food allergies. For these pathologies, jennet milk represents the best alternative to other types of milk. Therefore, jennet milk consumers are very sensible to the effects of pathogens' contaminations, and several hygienic practices during the milk production need to be adopted. During regular monitoring in one Sicilian jennet farm, Bacillus cereus in the milk was detected. In 3 bulk milk samples (maximum concentration: 1.2 x 103 ufc/ml, in 3 individual milk samples (10, 20 e 60 ufc/ml, in the milk filter (5 ufc/cm2, in the soil (maximum concentration: 1.5 x 103 ufc/g, on the hands and the gloves of two milkers, on the animal hide (from 1 to 3 ufc/cm2. No spores were detected. A total of 8 Bacillus cereus s.s. strains were analyzed for diarrhoic toxin, and 6 strains producing enterotoxins resulted. The improvement of environmental and milking hygienic conditions reduced Bacillus cereus concentration.

  13. Bioaccumulation of copper, zinc, cadmium and lead by Bacillus sp., Bacillus cereus, Bacillus sphaericus and Bacillus subtilis Bioacumulação de cobre, zinco, cádmio e chumbo por Bacillus sp., Bacillus cereus, Bacillus sphaericus e Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Antonio Carlos Augusto da Costa

    2001-03-01

    Full Text Available This work presents some results on the use of microbes from the genus Bacillus for uptake of cadmium, zinc, copper and lead ions. Maximum copper bioaccumulations were 5.6 mol/g biomass for B. sphaericus, 5.9 mol/g biomass for B. cereus and B. subtilis, and 6.4 mol/g biomass for Bacillus sp. Maximum zinc bioaccumulations were 4.3 mol/g biomass for B. sphaericus, 4.6 mol/g biomass for B. cereus, 4.8 mol/g biomass for Bacillus sp. and 5.0 mol/g biomass for B. subtilis. Maximum cadmium bioaccumulations were 8.0 mol/g biomass for B. cereus, 9.5 mol/g biomass for B. subtilis, 10.8 mol/g biomass for Bacillus sp. and 11.8 mol/g biomass for B. sphaericus. Maximum lead biomaccumulations were 0.7 mol/g biomass for B. sphaericus, 1.1 mol/g biomass for B. cereus, 1.4 mol/g biomass for Bacillus sp. and 1.8 mol/g biomass for B. subtilis. The different Bacillus strains tested presented distinct uptake capacities, and the best results were obtained for B. subtilis and B. cereus.Este trabalho apresenta resultados de acumulação dos íons metálicos cádmio, zinco, cobre e chumbo por bactérias do gênero Bacillus. A bioacumulação máxima de cobre foi 5,6 mol/g biomassa para B. sphaericus, 5,9 mol/g biomassa para B. cereus e B. subtilis, e 6,4 mol/g biomassa para Bacillus sp.. A bioacumulação máxima de zinco foi 4,3 mol/g biomassa para B. sphaericus, 4,6 mol/g biomassa para B. cereus, 4,8 mol/g biomassa para Bacillus sp. e 5,0 mol/g biomassa para B. subtilis. A bioacumulação máxima de cádmio foi 8,0 mol/g biomassa para B. cereus, 9,5 mol/g biomassa para B. subtilis, 10,8 mol/g biomassa para Bacillus sp. e 11,8 mol/g biomassa para B. sphaericus. A bioacumulação máxima de chumbo foi 0,7 mol/g biomassa para B. sphaericus, 1,1 mol/g biomassa para B. cereus, 1,4 mol/g biomassa para Bacillus sp. e 1,8 mol/g biomassa para B. subtilis. As distintas linhagens de Bacillus testadas apresentaram variáveis capacidades de carregamento de íons metálicos, sendo os

  14. Isolation of bacillus thuringiensis from different samples from Mansehra District

    International Nuclear Information System (INIS)

    The insecticidal activity of Bacillus thuringiensis has made it very interesting for the control of a variety of agricultural pests and human disease vectors. The present study is an attempt to explore the potential and diversity. of Bacillus thuringiensis. from the local environment for the control of cotton spotted bollworm (Earias sp.), a major pest of cotton. Two hundred and ninety eight samples of soil, grain dust, wild animal dung, birds dropping, decaying leaves and dead insects were collected from different ecological environments of Mansehra District yielding 438 Bacillus thuringiensis isolates that produce parasporal crystalline inclusions. In this study the soil samples were found to be the richest source for Bacillus thuringiensis. (author)

  15. Effect of oral administration of Bacillus coagulans B37 and Bacillus pumilus B9 strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model

    OpenAIRE

    Lopamudra Haldar; Gandhi, D.N.

    2016-01-01

    Aim: To investigate the effect of oral administration of two Bacillus strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model. Materials and Methods: An in vivo experiment was conducted for 49-day period on 36 adult male albino Wister rats divided equally into to four groups. After 7-day adaptation period, one group (T1) was fed on sterile skim milk along with basal diet for the next 28 days. Second (T2) and (T3) groups received spore biomass of Bacillus coagulans B...

  16. Emetic toxin-producing strains of Bacillus cereus show distinct characteristics within the Bacillus cereus group.

    NARCIS (Netherlands)

    Carlin, Frédéric; Fricker, Martina; Pielaat, Annemarie; Heisterkamp, Simon; Shaheen, Ranad; Salonen, Mirja Salkinoja; Svensson, Birgitta; Nguyen-the, Christophe; Ehling-Schulz, Monika

    2006-01-01

    One hundred representative strains of Bacillus cereus were selected from a total collection of 372 B. cereus strains using two typing methods (RAPD and FT-IR) to investigate if emetic toxin-producing hazardous B. cereus strains possess characteristic growth and heat resistance profiles. The strains

  17. Detection of Anthrax Simulants with Microcalorimetric Spectroscopy: Bacillus subtilis and Bacillus cereus Spores

    Science.gov (United States)

    Arakawa, Edward T.; Lavrik, Nickolay V.; Datskos, Panos G.

    2003-04-01

    Recent advances in the development of ultrasensitive micromechanical thermal detectors have led to the advent of novel subfemtojoule microcalorimetric spectroscopy (CalSpec). On the basis of principles of photothermal IR spectroscopy combined with efficient thermomechanical transduction, CalSpec provides acquisition of vibrational spectra of microscopic samples and absorbates. We use CalSpec as a method of identifying nanogram quantities of biological micro-organisms. Our studies focus on Bacillus subtilis and Bacillus cereus spores as simulants for Bacillus anthracis spores. Using CalSpec, we measured IR spectra of B. subtilis and B. cereus spores present on surfaces in nanogram quantities (approximately 100 -1000 spores). The spectra acquired in the wavelength range of 690 -4000 cm-1 (2.5 -14.5 μm) contain information-rich vibrational signatures that reflect the different ratios of biochemical makeup of the micro-organisms. The distinctive features in the spectra obtained for the two types of micro-organism can be used to distinguish between the spores of the Bacillus family. As compared with conventional IR and Fourier-transform IR microscopic spectroscopy techniques, the advantages of the present technique include significantly improved sensitivity (at least a full order of magnitude), absence of expensive IR detectors, and excellent potential for miniaturization.

  18. Complete Genome Sequence of Bacillus thuringiensis Bacteriophage Smudge.

    Science.gov (United States)

    Cornell, Jessica L; Breslin, Eileen; Schuhmacher, Zachary; Himelright, Madison; Berluti, Cassandra; Boyd, Charles; Carson, Rachel; Del Gallo, Elle; Giessler, Caris; Gilliam, Benjamin; Heatherly, Catherine; Nevin, Julius; Nguyen, Bryan; Nguyen, Justin; Parada, Jocelyn; Sutterfield, Blake; Tukruni, Muruj; Temple, Louise

    2016-01-01

    Smudge, a bacteriophage enriched from soil using Bacillus thuringiensis DSM-350 as the host, had its complete genome sequenced. Smudge is a myovirus with a genome consisting of 292 genes and was identified as belonging to the C1 cluster of Bacillus phages. PMID:27540049

  19. Complete Genome of Bacillus thuringiensis Myophage BigBertha

    OpenAIRE

    Ting, Jose H.; Smyth, Trinity B.; Chamakura, Karthik R.; Kuty Everett, Gabriel F.

    2013-01-01

    BigBertha is a myophage of Bacillus thuringiensis, a widely used biocontrol agent that is active against many insect pests of plants. Here, we present the complete annotated genome of BigBertha. The genome shares 85.9% sequence identity with Bacillus cereus phage B4.

  20. Non-peptide metabolites from the genus Bacillus.

    Science.gov (United States)

    Hamdache, Ahlem; Lamarti, Ahmed; Aleu, Josefina; Collado, Isidro G

    2011-04-25

    Bacillus species produce a number of non-peptide metabolites that display a broad spectrum of activity and structurally diverse bioactive chemical structures. Biosynthetic, biological, and structural studies of these metabolites isolated from Bacillus species are reviewed. This contribution also includes a detailed study of the activity of the metabolites described, especially their role in biological control mechanisms.

  1. Quantitative immunofluorescence studies of the serology of Bacillus anthracis spores.

    OpenAIRE

    Phillips, A. P.; Martin, K L

    1983-01-01

    A fluorescein-conjugated antibody against formalin-inactivated spores of Bacillus anthracis Vollum reacted only weakly with a variety of Bacillus species in microfluorometric immunofluorescence assays. A conjugated antibody against spores of B. anthracis Sterne showed little affinity for spores of several B. anthracis isolates including B. anthracis Vollum, indicating that more than one anthrax spore serotype exists.

  2. Complete Genome Sequence of Bacillus thuringiensis Bacteriophage Smudge

    Science.gov (United States)

    Cornell, Jessica L.; Breslin, Eileen; Schuhmacher, Zachary; Himelright, Madison; Berluti, Cassandra; Boyd, Charles; Carson, Rachel; Del Gallo, Elle; Giessler, Caris; Gilliam, Benjamin; Heatherly, Catherine; Nevin, Julius; Nguyen, Bryan; Nguyen, Justin; Parada, Jocelyn; Sutterfield, Blake; Tukruni, Muruj

    2016-01-01

    Smudge, a bacteriophage enriched from soil using Bacillus thuringiensis DSM-350 as the host, had its complete genome sequenced. Smudge is a myovirus with a genome consisting of 292 genes and was identified as belonging to the C1 cluster of Bacillus phages. PMID:27540049

  3. Complete Genome Sequence of Bacillus thuringiensis Bacteriophage Smudge.

    Science.gov (United States)

    Cornell, Jessica L; Breslin, Eileen; Schuhmacher, Zachary; Himelright, Madison; Berluti, Cassandra; Boyd, Charles; Carson, Rachel; Del Gallo, Elle; Giessler, Caris; Gilliam, Benjamin; Heatherly, Catherine; Nevin, Julius; Nguyen, Bryan; Nguyen, Justin; Parada, Jocelyn; Sutterfield, Blake; Tukruni, Muruj; Temple, Louise

    2016-08-18

    Smudge, a bacteriophage enriched from soil using Bacillus thuringiensis DSM-350 as the host, had its complete genome sequenced. Smudge is a myovirus with a genome consisting of 292 genes and was identified as belonging to the C1 cluster of Bacillus phages.

  4. Genetic map of the Bacillus stearothermophilus NUB36 chromosome

    Energy Technology Data Exchange (ETDEWEB)

    Vallier, H.; Welker, N.E. (Northwestern Univ., Evanston, IL (USA))

    1990-02-01

    A circular genetic map of Bacillus stearothermophilus NUB36 was constructed by transduction with bacteriophage TP-42C and protoplast fusion. Sixty-four genes were tentatively assigned a cognate Bacillus subtilis gene based on growth response to intermediates or end products of metabolism, cross-feeding, accumulation of intermediates, or their relative order in a linkage group. Although the relative position of many genes on the Bacillus subtilis genetic map appears to be similar, some differences were detected. The tentative order of the genes in the Bacillus stearothermophilus aro region is aspB-aroBAFEC-tyra-hisH-(trp), whereas it is aspB-aroE-tyrA-hisH-(trp)-aroHBF in Bacillus subtilis. The aroA, aroC, and aroG genes in Bacillus subtilis are located in another region. The tentative order of genes in the trp operon of Bacillus stearothermophilus is trpFCDABE, whereas it is trpABFCDE in Bacillus subtilis.

  5. Bacillus subtilis regulatory protein GerE

    OpenAIRE

    Ducros, V M A; Brannigan, J.A.; Lewis, R J; Wilkinson, A.J.

    1998-01-01

    GerE is the latest-acting of a series of factors which regulate gene expression in the mother cell during sporulation in Bacillus. The gene encoding GerE has been cloned from B. subtilis and overexpressed in Escherichia coli. Purified GerE has been crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. The small plate-like crystals belong to the monoclinic space group C2 and diffract beyond 2.2 Angstrom resolution with a synchrotron radiation X-ra...

  6. Enhanced transformation efficiency of recalcitrant Bacillus cereus and Bacillus weihenstephanensis isolates upon in vitro methylation of plasmid DNA

    NARCIS (Netherlands)

    Nierop Groot, M.N.; Nieboer, F.; Abee, T.

    2008-01-01

    Digestion patterns of chromosomal DNAs of Bacillus cereus and Bacillus weihenstephanensis strains suggest that Sau3AI-type restriction modification systems are widely present among the isolates tested. In vitro methylation of plasmid DNA was used to enhance poor plasmid transfer upon electroporation

  7. ABILITY OF BACTERIAL CONSORTIUM: Bacillus coagulans, Bacilus licheniformis, Bacillus pumilus, Bacillus subtilis, Nitrosomonas sp. and Pseudomonas putida IN BIOREMEDIATION OF WASTE WATER IN CISIRUNG WASTE WATER TREATMENT PLANT

    Directory of Open Access Journals (Sweden)

    Ratu SAFITRI

    2015-10-01

    Full Text Available This study was conducted in order to determine the ability of bacterial consortium: Bacillus coagulans, Bacilus licheniformis, Bacillus pumilus, Bacillus subtilis, Nitrosomonas sp., and Pseudomonas putida in bioremediation of wastewater origin Cisirung WWTP. This study uses an experimental method completely randomized design (CRD, which consists of two treatment factors (8x8 factorial design. The first factor is a consortium of bacteria (K, consisting of 8 level factors (k1, k2, k3, k4, k5, k6, k7, and k8. The second factor is the time (T, consisting of a 7 level factors (t0, t1, t2, t3, t4, t5, t6, and t7. Test parameters consist of BOD (Biochemical Oxygen Demand, COD (Chemical Oxygen Demand, TSS (Total Suspended Solid, Ammonia and Population of Microbes during bioremediation. Data were analyzed by ANOVA, followed by Duncan test. The results of this study showed that the consortium of Bacillus pumilus, Bacillus subtilis, Bacillus coagulans, Nitrosomonas sp., and Pseudomonas putida with inoculum concentration of 5% (k6 is a consortium of the most effective in reducing BOD 71.93%, 64.30% COD, TSS 94.85%, and 88.58% of ammonia.

  8. The Phylogeny of Bacillus cereus sensu lato.

    Science.gov (United States)

    Okinaka, Richard T; Keim, Paul

    2016-02-01

    The three main species of the Bacillus cereus sensu lato, B. cereus, B. thuringiensis, and B. anthracis, were recognized and established by the early 1900s because they each exhibited distinct phenotypic traits. B. thuringiensis isolates and their parasporal crystal proteins have long been established as a natural pesticide and insect pathogen. B. anthracis, the etiological agent for anthrax, was used by Robert Koch in the 19th century as a model to develop the germ theory of disease, and B. cereus, a common soil organism, is also an occasional opportunistic pathogen of humans. In addition to these three historical species designations, are three less-recognized and -understood species: B. mycoides, B. weihenstephanensis, and B. pseudomycoides. All of these "species" combined comprise the Bacillus cereus sensu lato group. Despite these apparently clear phenotypic definitions, early molecular approaches to separate the first three by various DNA hybridization and 16S/23S ribosomal sequence analyses led to some "confusion" because there were limited differences to differentiate between these species. These and other results have led to frequent suggestions that a taxonomic change was warranted to reclassify this group to a single species. But the pathogenic properties of B. anthracis and the biopesticide applications of B. thuringiensis appear to "have outweighed pure taxonomic considerations" and the separate species categories are still being maintained. B. cereus sensu lato represents a classic example of a now common bacterial species taxonomic quandary. PMID:26999390

  9. Fast Neutron Radiation Effects on Bacillus Subtili

    Institute of Scientific and Technical Information of China (English)

    CHEN Xiaoming; REN Zhenglong; ZHANG Jianguo; ZHENG Chun; TAN Bisheng; YANG Chengde; CHU Shijin

    2009-01-01

    To examine the sterilizing effect and mechanism of neutron radiation, Bacillus sub-tilis vat. niger, strain (ATCC 9372) spores were irradiated with the fast neutron from the Chinese fast burst reactor Ⅱ(CFBR-Ⅱ). The plate-count results indicated that the D10 value was 384.6 Gy with a neutron radiation dose rate of 7.4 Gy/min. The rudimental catalase activity of the spores declined obviously with the increase in the radiation dose. Meanwhile, under the scanning electron microscope, no visible influence of the neutron radiation on the spore configuration was detected even if the dose was increased to 4 kGy. The content and distribution of DNA double-strand breaks induced by neutron radiation at different doses were measured and quantified by pulsed-field gel electrophoresis (PFGE). Further analysis of the DNA release percentage (PR), the DNA breakage level (L), and the average molecular weight, indicated that DNA fragments were obvi-ously distributed around the 5 kb regions at different radiation doses, which suggests that some points in the DNA molecule were sensitive to neutron radiation. Both PR and L varied regularly to some extent with the increase in radiation dose. Thus neutron radiation has a high sterilization power, and can induce falling enzyme activity and DNA breakage in Bacillus subtilis spores

  10. ISOLATION AND CHARACTERIZATION OF CRUDE OIL DEGRADING BACILLUS SPP.

    Directory of Open Access Journals (Sweden)

    A. Akhavan Sepahi, I. Dejban Golpasha, M. Emami, A. M. Nakhoda

    2008-07-01

    Full Text Available Today, application of microorganisms for removing crude oil pollution from contaminated sites as bioremediation studies, was considered by scientists because other methods such as surfactant washing and incineration lead to production of more toxic compounds and they are non-economic. Fifteen crude oil degrading bacillus spp. were isolated from contaminated sites. Two isolated showed best growth in liquid media with 1-3% (v/v crude oil and mineral salt medium, then studied for enzymatic activities on tested media. The results showed maximal increase in optical densities and total viable count concomitant with decrease in pH on fifth day of experimental period for bacillus S6. Typical generation time on mineral salt with 1% crude oil is varying between 18-20h, 25-26h respectively for bacillus S6 and S35. Total protein was monitored at determined time intervals as biodegradation indices. Increasing of protein concentration during the incubation period reveals that isolated bacillus can degrade crude oil and increase microbial biomass. These bacillus spp. reduced surface tension from 60 (mN/m to 31 and 38 (mN/m, It means that these bacillus spp. can produce sufficient surfactant and have good potential of emulsification capacity. The results demonstrated that these bacillus spp. can utilize crude oil as a carbon and energy source.

  11. Occurrence and significance of Bacillus cereus and Bacillus thuringiensis in ready-to-eat food

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Ørum-Smidt, Lasse; Andersen, Sigrid R;

    2005-01-01

    had at least one gene or component involved in human diarrhoeal disease, while emetic toxin was related to only one B. cereus strain. A new observation was that 31 out of the 40 randomly selected B. cereus-like strains could be classified as Bacillus thuringiensis due to crystal production and......Among 48,901 samples of ready-to-eat food products at the Danish retail market, 0.5% had counts of Bacillus cereus-like bacteria above 10(4) cfu g(-1). The high counts were most frequently found in starchy, cooked products, but also in fresh cucumbers and tomatoes. Forty randomly selected strains....../or content of cry genes. Thus, a large proportion of the B. cereus-like organisms present in food may belong to B. thuringiensis....

  12. Expression, purification, and characterization of a thermophilic neutral protease from Bacillus stearothermophilus in Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The gene coding for a thermophilic neutral protease from Bacillus stearothermophilus was expressed in Bacillus subtilis DB104, under the control of the sacB gene promoter. This was followed by either the native signal peptide sequence of this protease or the signal peptide sequence of the sacB gene. The protease was purified 3.8-fold, with a specific activity of 16530 U mg-1. As analyzed by SDS-PAGE, the molecular mass of the expressed protease was about 35 kDa, and the optimal temperature and pH of the protease were 65℃ and 7.5, respectively. Moreover, it still had about 80% activity after 1 h reaction at 65℃.

  13. Expression, purification, and characterization of a thermophilic neutral protease from Bacillus stearothermophilus in Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The gene coding for a thermophilic neutral protease from Bacillus stearothermophilus was expressed in Bacillus subtilis DB104, under the control of the sacB gene promoter. This was followed by either the native signal peptide sequence of this protease or the signal peptide sequence of the sacB gene. The protease was purified 3.8-fold, with a specific activity of 16530 U mg-1. As analyzed by SDS-PAGE, the molecular mass of the expressed protease was about 35 kDa, and the optimal temperature and pH of the protease were 65℃ and 7.5, respectively. Moreover, it still had about 80% activity after 1 h reaction at 65 ℃ .

  14. An Optical Biosensor for Bacillus Cereus Spore Detection

    Science.gov (United States)

    Li, Chengquan; Tom, Harry W. K.

    2005-03-01

    We demonstrate a new transduction scheme for optical biosensing. Bacillus cereus is a pathogen that may be found in food and dairy products and is able to produce toxins and cause food poisoning. It is related to Bacillus anthracis (anthrax). A CCD array covered with micro-structured glass coverslip is used to detect the optical resonant shift due to the binding of the antigen (bacillus cereus spore) to the antibody (polyclonal antibody). This novel optical biosensor scheme has the potential for detecting 10˜100 bioagents in a single device as well as the potential to test for antigens with multiple antibody tests to avoid ``false positives.''

  15. Flow-cytometric Analysis of Bacillus anthracis Spores

    Directory of Open Access Journals (Sweden)

    D. V. Kamboj

    2006-11-01

    Full Text Available Flow-cytometric technique has been established as a powerful tool for detection andidentification of microbiological agents. Unambiguous and rapid detection of Bacillus anthracisspores has been reported using immunoglobulin G-fluorescein isothiocyanate conjugate againstlive spores. In addition to the high sensitivity, the present technique could differentiate betweenspores of closely related species, eg, Bacillus cereus and Bacillus subtilis using fluorescenceintensity. The technique can be used for detection of live as well as inactivated spores makingit more congenial for screening of suspected samples of bioterrorism.

  16. Genetic Differentiation between Sympatric Populations of Bacillus cereus and Bacillus thuringiensis

    Science.gov (United States)

    Vilas-Boas, Gislayne; Sanchis, Vincent; Lereclus, Didier; Lemos, Manoel Victor F.; Bourguet, Denis

    2002-01-01

    Little is known about genetic exchanges in natural populations of bacteria of the spore-forming Bacillus cereus group, because no population genetics studies have been performed with local sympatric populations. We isolated strains of Bacillus thuringiensis and B. cereus from small samples of soil collected at the same time from two separate geographical sites, one within the forest and the other at the edge of the forest. A total of 100 B. cereus and 98 B. thuringiensis strains were isolated and characterized by electrophoresis to determine allelic composition at nine enzymatic loci. We observed genetic differentiation between populations of B. cereus and B. thuringiensis. Populations of a given Bacillus species—B. thuringiensis or B. cereus—were genetically more similar to each other than to populations of the other Bacillus species. Hemolytic activity provided further evidence of this genetic divergence, which remained evident even if putative clones were removed from the data set. Our results suggest that the rate of gene flow was higher between strains of the same species, but that exchanges between B. cereus and B. thuringiensis were nonetheless possible. Linkage disequilibrium analysis revealed sufficient recombination for B. cereus populations to be considered panmictic units. In B. thuringiensis, the balance between clonal proliferation and recombination seemed to depend on location. Overall, our data indicate that it is not important for risk assessment purposes to determine whether B. cereus and B. thuringiensis belong to a single or two species. Assessment of the biosafety of pest control based on B. thuringiensis requires evaluation of the extent of genetic exchange between strains in realistic natural conditions. PMID:11872495

  17. Bacillus subtilis as potential producer for polyhydroxyalkanoates

    Directory of Open Access Journals (Sweden)

    Patel Sanjay KS

    2009-07-01

    Full Text Available Abstract Polyhydroxyalkanoates (PHAs are biodegradable polymers produced by microbes to overcome environmental stress. Commercial production of PHAs is limited by the high cost of production compared to conventional plastics. Another hindrance is the brittle nature and low strength of polyhydroxybutyrate (PHB, the most widely studied PHA. The needs are to produce PHAs, which have better elastomeric properties suitable for biomedical applications, preferably from inexpensive renewable sources to reduce cost. Certain unique properties of Bacillus subtilis such as lack of the toxic lipo-polysaccharides, expression of self-lysing genes on completion of PHA biosynthetic process – for easy and timely recovery, usage of biowastes as feed enable it to compete as potential candidate for commercial production of PHA.

  18. Bacillus circulans exopolysaccharide: Production, characterization and bioactivities.

    Science.gov (United States)

    Vidhyalakshmi, R; Valli, Nachiyar C; Narendra Kumar, G; Sunkar, Swetha

    2016-06-01

    A bacterium with the ability to produce appreciable amount of exopolysaccharide was isolated from slimy layer of coconut. 16S rDNA analysis identified the organism as Bacillus circulans. EPS production was observed at all stages of culture growth and reached maximum of 0.065mg/ml by 96h, which on further incubation started to decrease. Response Surface Methodology using Box Behnken design has shown the influence of sucrose which was found to be directly proportional to exopolysaccharide production with production reaching 1.09mg/ml. HPLC analysis identified the presence of glucose, mannose, fructose and verbascose and NMR analysis confirmed the presence of glucose, mannose and galactose. Even though the extracted B. circulans EPS did not show appreciable anti-bacterial or anti-fungal activity, it exhibited appreciable antioxidant, anti-inflammatory and anti-tumor activity. PMID:26902891

  19. Characterization of Bacillus anthracis persistence in vivo.

    Directory of Open Access Journals (Sweden)

    Sarah A Jenkins

    Full Text Available Pulmonary exposure to Bacillus anthracis spores initiates inhalational anthrax, a life-threatening infection. It is known that dormant spores can be recovered from the lungs of infected animals months after the initial spore exposure. Consequently, a 60-day course antibiotic treatment is recommended for exposed individuals. However, there has been little information regarding details or mechanisms of spore persistence in vivo. In this study, we investigated spore persistence in a mouse model. The results indicated that weeks after intranasal inoculation with B. anthracis spores, substantial amounts of spores could be recovered from the mouse lung. Moreover, spores of B. anthracis were significantly better at persisting in the lung than spores of a non-pathogenic Bacillus subtilis strain. The majority of B. anthracis spores in the lung were tightly associated with the lung tissue, as they could not be readily removed by lavage. Immunofluorescence staining of lung sections showed that spores associated with the alveolar and airway epithelium. Confocal analysis indicated that some of the spores were inside epithelial cells. This was further confirmed by differential immunofluorescence staining of lung cells harvested from the infected lungs, suggesting that association with lung epithelial cells may provide an advantage to spore persistence in the lung. There was no or very mild inflammation in the infected lungs. Furthermore, spores were present in the lung tissue as single spores rather than in clusters. We also showed that the anthrax toxins did not play a role in persistence. Together, the results suggest that B. anthracis spores have special properties that promote their persistence in the lung, and that there may be multiple mechanisms contributing to spore persistence.

  20. Genome Sequence of Bacillus thuringiensis subsp. kurstaki Strain HD-1

    OpenAIRE

    Day, Michael; Ibrahim, Mohamed; Dyer, David; Bulla, Lee

    2014-01-01

    We report here the complete genome sequence of Bacillus thuringiensis subsp. kurstaki strain HD-1, which serves as the primary U.S. reference standard for all commercial insecticidal formulations of B. thuringiensis manufactured around the world.

  1. Effects of probiotic Bacillus species in aquaculture – An overview

    Directory of Open Access Journals (Sweden)

    Cristian-Teodor BURUIANĂ

    2014-12-01

    Full Text Available The ingestion of a large amount of certain types of beneficial bacteria can reduce the multiplication and development of pathogenic bacteria in the gut. A “probiotic” is a product that contains live microorganisms which positively influence the host intestinal microbiota by preventing the proliferation of pathogenic bacteria and promoting the growth and development of beneficial bacteria. Bacillus spp. are Gram-positive endospore-forming bacteria with beneficial effects in aquaculture industry. The dietary supplementation of Bacillus spp. in fish culture improved especially growth performance, immune response and the disease resistance of fish against pathogenic bacterial infections. The objective of the current paper is to review the recent published investigations reported in the scientific literature on the use of probiotic Bacillus spp. in aquaculture, focusing on their beneficial effects on the host. This review includes the main effects of Bacillus spp. administration in shrimp culture, carp culture, tilapia culture, and other fish culture.

  2. Two Genes Encoding Uracil Phosphoribosyltransferase Are Present in Bacillus subtilis

    DEFF Research Database (Denmark)

    Martinussen, Jan; Glaser, Philippe; Andersen, Paal S.;

    1995-01-01

    Uracil phosphoribosyltransferase (UPRTase) catalyzes the key reaction in the salvage of uracil in many microorganisms. Surprisingly, two genes encoding UPRTase activity were cloned from Bacillus subtilis by complementation of an Escherichia coli mutant. The genes were sequenced, and the putative...

  3. The Complete Genome Sequence of Bacillus thuringiensis AlHakam

    Energy Technology Data Exchange (ETDEWEB)

    Challacombe, Jean F.; Altherr, Michael R.; Xie, Gary; Bhotika,Smriti S.; Brown, Nancy; Bruce, David; Campbell, Connie S.; Campbell,Mary L.; Chen, Jin; Chertkov, Olga; Cleland, Cathy; Dimitrijevic, Mira; Doggett, Norman A.; Fawcett, John J.; Glavina, Tijana; Goodwin, Lynne A.; Green, Lance D.; Han, Cliff S.; Hill, Karen K.; Hitchcock, Penny; Jackson, Paul J.; Keim, Paul; Kewalramani, Avinash Ramesh; Longmire, Jon; Lucas, Susan; Malfatti, Stephanie; Martinez, Diego; McMurry, Kim; Meincke, Linda J.; Misra, Monica; Moseman, Bernice L.; Mundt, Mark; Munk,A. Christine; Okinaka, Richard T.; Parson-Quintana, B.; Reilly, LeePhilip; Richardson, Paul; Robinson, Donna L.; Rubin, Eddy; Saunders,Elizabeth; Tapia, Roxanne; Tesmer, Judith G.; Thayer, Nina; Thompson,Linda S.; Tice, Hope; Ticknor, Lawrence O.; Wills, Patti L.; Gilna, Paul; Brettin, Thomas S.

    2007-04-01

    Bacillus thuringiensis is an insect pathogen that is widelyused as a biopesticide (3). Here we report the finished, annotated genomesequence of B. thuringiensis Al Hakam, which was collected in Iraq by theUnited Nations Special Commission (2).

  4. BOOK REVIEW: BACILLUS THURINGIENSIS: A CORNERSTONE OF MODERN AGRICULTURE

    Science.gov (United States)

    Are you interested in the technical issues surrounding the use of Bacillus thuringiensis pesticidal traits as sprays and as plant incorporated protectants (transgenic crops)? Should the dimensions of human health, ecology, entomology, risk assessment, resistance management, and d...

  5. Antifungal activity of Bacillus sp. isolated from compost.

    Science.gov (United States)

    Czaczyk, K; Stachowiak, B; Trojanowska, K; Gulewicz, K

    2000-01-01

    Four strains of Bacillus isolated from lupine compost exhibited an antifungal activity against six plant fungal pathogens (Rhizoctonia solani, Bipolaris sorokiniana, Sclerotinia sclerotiorum, Trichothecium roseum, Fusarium solani, Fusarium oxysporum). It was significantly influenced by the composition of the cultivation media.

  6. Selection of Bacillus subtilis mutants impaired in ammonia assimilation.

    OpenAIRE

    Dean, D R; Aronson, A I

    1980-01-01

    The selection of Bacillus subtilis mutants capable of using D-histidine to fulfill a requirement for L-histidine resulted in mutants with either no glutamate synthase activity or increased amounts of an altered glutamine synthetase.

  7. Protein engineering of cyclodextrin glycosyltransferase from Bacillus circulans strain 251

    NARCIS (Netherlands)

    Penninga, Dirk

    1996-01-01

    An enormous diversity of molecular functions in living organisms is carried out by proteins. Our studies have focussed on the functional analysis of a starch-converting enzyme, cyclodextrin glycosyltransferase (CGTase) from Bacillus circulans strain 251. Zie: Summary

  8. Regulation of cry Gene Expression in Bacillus thuringiensis

    OpenAIRE

    Chao Deng; Qi Peng; Fuping Song; Didier Lereclus

    2014-01-01

    Bacillus thuringiensis differs from the closely related Bacillus cereus group species by its ability to produce crystalline inclusions. The production of these crystals mainly results from the expression of the cry genes, from the stability of their transcripts and from the synthesis, accumulation and crystallization of large amounts of insecticidal Cry proteins. This process normally coincides with sporulation and is regulated by various factors operating at the transcriptional, post-transcr...

  9. Natural Dissemination of Bacillus anthracis Spores in Northern Canada

    OpenAIRE

    Dragon, D C; Bader, D. E.; Mitchell, J.; Woollen, N.

    2005-01-01

    Soil samples were collected from around fresh and year-old bison carcasses and areas not associated with known carcasses in Wood Buffalo National Park during an active anthrax outbreak in the summer of 2001. Sample selection with a grid provided the most complete coverage of a site. Soil samples were screened for viable Bacillus anthracis spores via selective culture, phenotypic analysis, and PCR. Bacillus anthracis spores were isolated from 28.4% of the samples. The highest concentrations of...

  10. Genetic analysis of petrobactin transport in Bacillus anthracis

    OpenAIRE

    Carlson, Paul E.; Dixon, Shandee D.; Janes, Brian K.; Carr, Katherine A.; Nusca, Tyler D.; Anderson, Erica C.; Keene, Sarra E.; Sherman, David H.; Hanna, Philip C.

    2010-01-01

    Iron acquisition mechanisms play an important role in the pathogenesis of many infectious microbes. In Bacillus anthracis, the siderophore petrobactin is required for both growth in iron depleted conditions and for full virulence of the bacterium. Here we demonstrate the roles of two putative petrobactin binding proteins FatB and FpuA (encoded by GBAA5330 and GBAA4766, respectively) in Bacillus anthracis iron acquisition and pathogenesis. Markerless deletion mutants were created using allelic...

  11. Expression of UGA-Containing Mycoplasma Genes in Bacillus subtilis

    OpenAIRE

    Kannan, T. R.; Baseman, Joel B.

    2000-01-01

    We used Bacillus subtilis to express UGA-containing Mycoplasma genes encoding the P30 adhesin (one UGA) of Mycoplasma pneumoniae and methionine sulfoxide reductase (two UGAs) of Mycoplasma genitalium. Due to natural UGA suppression, these Mycoplasma genes were expressed as full-length protein products, but at relatively low efficiency, in recombinant wild-type Bacillus. The B. subtilis-expressed Mycoplasma proteins appeared as single bands and not as multiple bands compared to expression in r...

  12. Laser-induced speckle scatter patterns in Bacillus colonies

    Directory of Open Access Journals (Sweden)

    Huisung eKim

    2014-10-01

    Full Text Available Label-free bacterial colony phenotyping technology called BARDOT (BActerial Rapid Detection using Optical scattering Technology provided successful classification of several different bacteria at the genus, species, and serovar level. Recent experiments with colonies of Bacillus species provided strikingly different characteristics of elastic light scatter (ELS patterns, which were comprised of random speckles compared to other bacteria, which are dominated by concentric rings and spokes. Since this laser-based optical sensor interrogates the whole volume of the colony, 3-D information of micro- and macro-structures are all encoded in the far-field scatter patterns. Here, we present a theoretical model explaining the underlying mechanism of the speckle formation by the colonies from Bacillus species. Except for Bacillus polymyxa, all Bacillus spp. produced random bright spots on the imaging plane, which presumably dependent on the cellular and molecular organization and content within the colony. Our scatter model-based analysis revealed that colony spread resulting in variable surface roughness can modify the wavefront of the scatter field. As the center diameter of the Bacillus spp. colony grew from 500 μm to 900 μm, average speckles area decreased 2-fold and the number of small speckles increased 7-fold. In conclusion, as Bacillus colony grows, the average speckle size in the scatter pattern decreases and the number of smaller speckle increases due to the swarming growth characteristics of bacteria within the colony.

  13. Transfer of the toxin protein genes of Bacillus sphaericus into Bacillus thuringiensis subsp. israelensis and their expression.

    OpenAIRE

    Bourgouin, C.; Delécluse, A; La Torre, F.; Szulmajster, J

    1990-01-01

    The genes encoding the toxic determinants of Bacillus sphaericus have been expressed in a nontoxic and a toxic strain of Bacillus thuringiensis subsp. israelensis. In both cases, the B. sphaericus toxin proteins were produced at a high level during sporulation of B. thuringiensis and accumulated as crystalline structures. B. thuringiensis transformants expressing B. sphaericus and B. thuringiensis subsp. israelensis toxins did not show a significant enhancement of toxicity against Aedes aegyp...

  14. Ground Anthrax Bacillus Refined Isolation (GABRI) method for analyzing environmental samples with low levels of Bacillus anthracis contamination

    OpenAIRE

    Fasanella, Antonio; Di Taranto, Pietro; Garofolo, Giuliano; Colao, Valeriana; Marino, Leonardo; Buonavoglia, Domenico; Pedarra, Carmine; Adone, Rosanna; Hugh-Jones, Martin

    2013-01-01

    Background In this work are reported the results of a qualitative analytical method capable of detecting Bacillus anthracis spores when they are present in very low concentration in the soil. The Ground Anthrax Bacillus Refined Isolation (GABRI) method, assessed in our laboratory, was compared with the classic method. The comparison involved artificially anthrax-contaminated soil samples (500 spores/7.5 grams soil) and naturally contaminated soil samples collected in Bangladesh during a field...

  15. Comparative sequence analyses on the 16S rRNA (rDNA) of Bacillus acidocaldarius, Bacillus acidoterrestris, and Bacillus cycloheptanicus and proposal for creation of a new genus, Alicyclobacillus gen. nov

    Science.gov (United States)

    Wisotzkey, J. D.; Jurtshuk, P. Jr; Fox, G. E.; Deinhard, G.; Poralla, K.

    1992-01-01

    Comparative 16S rRNA (rDNA) sequence analyses performed on the thermophilic Bacillus species Bacillus acidocaldarius, Bacillus acidoterrestris, and Bacillus cycloheptanicus revealed that these organisms are sufficiently different from the traditional Bacillus species to warrant reclassification in a new genus, Alicyclobacillus gen. nov. An analysis of 16S rRNA sequences established that these three thermoacidophiles cluster in a group that differs markedly from both the obligately thermophilic organisms Bacillus stearothermophilus and the facultatively thermophilic organism Bacillus coagulans, as well as many other common mesophilic and thermophilic Bacillus species. The thermoacidophilic Bacillus species B. acidocaldarius, B. acidoterrestris, and B. cycloheptanicus also are unique in that they possess omega-alicylic fatty acid as the major natural membranous lipid component, which is a rare phenotype that has not been found in any other Bacillus species characterized to date. This phenotype, along with the 16S rRNA sequence data, suggests that these thermoacidophiles are biochemically and genetically unique and supports the proposal that they should be reclassified in the new genus Alicyclobacillus.

  16. Inactivation of Bacillus atrophaeus by OH radicals

    Science.gov (United States)

    Ono, Ryo; Yonetamari, Kenta; Tokumitsu, Yusuke; Yonemori, Seiya; Yasuda, Hachiro; Mizuno, Akira

    2016-08-01

    The inactivation of Bacillus atrophaeus by OH radicals is measured. This study aims to evaluate the bactericidal effects of OH radicals produced by atmospheric-pressure nonthermal plasma widely used for plasma medicine; however, in this study, OH radicals are produced by vacuum ultraviolet (VUV) photolysis of water vapor instead of plasma to allow the production of OH radicals with almost no other reactive species. A 172 nm VUV light from a Xe2 excimer lamp irradiates a He–H2O mixture flowing in a quartz tube to photodissociate H2O to produce OH, H, O, HO2, H2O2, and O3. The produced reactive oxygen species (ROS) flow out of the quartz tube nozzle to the bacteria on an agar plate and cause inactivation. The inactivation by OH radicals among the six ROS is observed by properly setting the experimental conditions with the help of simulations calculating the ROS densities. A 30 s treatment with approximately 0.1 ppm OH radicals causes visible inactivation.

  17. Bacillus anthracis diversity in Kruger National Park.

    Science.gov (United States)

    Smith, K L; DeVos, V; Bryden, H; Price, L B; Hugh-Jones, M E; Keim, P

    2000-10-01

    The Kruger National Park (KNP), South Africa, has a recorded history of periodic anthrax epidemics causing widespread disease among wild animals. Bacillus anthracis is the causative agent of anthrax, a disease primarily affecting ungulate herbivores. Worldwide there is little diversity among B. anthracis isolates, but examination of variable-number tandem repeat (VNTR) loci has identified six major clones, with the most dissimilar types split into the A and B branches. Both the A and B types are found in southern Africa, giving this region the greatest genetic diversity of B. anthracis worldwide. Consequently, southern Africa has been hypothesized to be the geographic origin of B. anthracis. In this study, we identify the genotypic types of 98 KNP B. anthracis isolates using multiple-locus VNTR analysis. Two major types are evident, the A branch and the B branch. The spatial and temporal distribution of the different genotypes indicates that anthrax epidemic foci are independent, though correlated through environmental cues. Kruger B isolates were found on significantly higher-calcium and higher-pH soils than were Kruger type A. This relationship between genotype and soil chemistry may be due to adaptive differences among divergent anthrax strains. While this association may be simply fortuitous, adaptation of A types to diverse environmental conditions is consistent with their greater geographic dispersal and genetic dissimilarity.

  18. Cannibalism stress response in Bacillus subtilis.

    Science.gov (United States)

    Höfler, Carolin; Heckmann, Judith; Fritsch, Anne; Popp, Philipp; Gebhard, Susanne; Fritz, Georg; Mascher, Thorsten

    2016-01-01

    When faced with carbon source limitation, the Gram-positive soil organism Bacillus subtilis initiates a survival strategy called sporulation, which leads to the formation of highly resistant endospores that allow B. subtilis to survive even long periods of starvation. In order to avoid commitment to this energy-demanding and irreversible process, B. subtilis employs another strategy called 'cannibalism' to delay sporulation as long as possible. Cannibalism involves the production and secretion of two cannibalism toxins, sporulation delaying protein (SDP) and sporulation killing factor (SKF), which are able to lyse sensitive siblings. The lysed cells are thought to then provide nutrients for the cannibals to slow down or even prevent them from entering sporulation. In this study, we uncovered the role of the cell envelope stress response (CESR), especially the Bce-like antimicrobial peptide detoxification modules, in the cannibalism stress response during the stationary phase. SDP and SKF specifically induce Bce-like systems and some extracytoplasmic function σ factors in stationary-phase cultures, but only the latter provide some degree of protection. A full Bce response is only triggered by mature toxins, and not by toxin precursors. Our study provides insights into the close relationship between stationary-phase survival and the CESR of B. subtilis. PMID:26364265

  19. Comparison of different Bacillus subtilis expression systems.

    Science.gov (United States)

    Vavrová, Ludmila; Muchová, Katarína; Barák, Imrich

    2010-11-01

    Bacillus subtilis is considered to have great potential as a host for the production and secretion of recombinant proteins. Many different expression systems have been developed for B. subtilis. Here we compare two widely used expression systems, the IPTG-inducible derivative of spac system (hyper-spank) and the xylose-inducible (xyl) to the SURE (subtilin-regulated gene expression) system. Western blot analysis of the membrane protein SpoIISA together with its protein partner SpoIISB showed that the highest expression level of this complex is obtained using the SURE system. Measurement of β-galactosidase activities of the promoter-lacZ fusions in individual expression systems confirmed that the P(spaS) promoter of the SURE system is the strongest of those compared, although the induction/repression ratio reached only 1.84. Based on these results, we conclude that the SURE system is the most efficient of these three B. subtilis expression systems in terms of the amount of expressed product. Remarkably, the yield of the SpoIISA-SpoIISB complex obtained from B. subtilis was comparable to that normally obtained from the Escherichia coli arabinose-inducible expression system. PMID:20863884

  20. Bacillus thuringiensis Conjugation in Simulated Microgravity

    Science.gov (United States)

    Beuls, Elise; van Houdt, Rob; Leys, Natalie; Dijkstra, Camelia; Larkin, Oliver; Mahillon, Jacques

    2009-10-01

    Spaceflight experiments have suggested a possible effect of microgravity on the plasmid transfer among strains of the Gram-positive Bacillus thuringiensis, as opposed to no effect recorded for Gram-negative conjugation. To investigate these potential effects in a more affordable experimental setup, three ground-based microgravity simulators were tested: the Rotating Wall Vessel (RWV), the Random Positioning Machine (RPM), and a superconducting magnet. The bacterial conjugative system consisted in biparental matings between two B. thuringiensis strains, where the transfer frequencies of the conjugative plasmid pAW63 and its ability to mobilize the nonconjugative plasmid pUB110 were assessed. Specifically, potential plasmid transfers in a 0-g position (simulated microgravity) were compared to those obtained under 1-g (normal gravity) condition in each device. Statistical analyses revealed no significant difference in the conjugative and mobilizable transfer frequencies between the three different simulated microgravitational conditions and our standard laboratory condition. These important ground-based observations emphasize the fact that, though no stimulation of plasmid transfer was observed, no inhibition was observed either. In the case of Gram-positive bacteria, this ability to exchange plasmids in weightlessness, as occurs under Earth's conditions, should be seen as particularly relevant in the scope of spread of antibiotic resistances and bacterial virulence.

  1. A pangenomic study of Bacillus thuringiensis

    Institute of Scientific and Technical Information of China (English)

    Yongjun Fang; Songnian Hu; Jie Zhang; Ibrahim A1-Mssallem; Jun Yu; Zhaolong Li; Jiucheng Liu; Changlong Shu; Xumin Wang; Xiaowei Zhang; Xiaoguang Yu; Duojun Zhao; Guiming Liu

    2011-01-01

    Bacillus thuringiensis (B.thuringiensis) is a soil-dwelling Gram-positive bacterium and its plasmid-encoded toxins (Cry) are commonly used as biological alternatives to pesticides.In a pangenomic study,we sequenced seven B.thuringiensis isolates in both high coverage and base quality using the next-generation sequencing platform.The B.thuringiensis pangenome was extrapolated to have 4196 core genes and an asymptotic value of 558 unique genes when a new genome is added.Compared to the pangenomes of its closely related species of the same genus,B.thuringiensis pangenome shows an open characteristic,similar to B.cereus but not to B.anthracis; the latter has a closed pangenome.We also found extensive divergence among the seven B.thuringiensis genome assemblies,which harbor ample repeats and single nucleotide polymorphisms (SNPs).The identities among orthologous genes are greater than 84.5% and the hotspots for the genome variations were discovered in genomic regions of 2.3-2.8 Mb and 5.0-5.6 Mb.We concluded that high-coverage sequence assemblies from multiple strains,before all the gaps are closed,are very useful for pangenomic studies.

  2. Surface topography of the Bacillus stearothermophilus ribosome

    International Nuclear Information System (INIS)

    The surface topography of the intact 70S ribosome and free 30S and 50S subunits from Bacillus stearothermophilus strain 2,184 was investigated by lactoperoxidase-catalyzed iodination. Two-dimensional polyacrylamide gel electrophoresis was employed to separate ribosomal proteins for analysis of their reactivity. Free 50S subunits incorporated about 18% more 125I than did 50S subunits derived from 70S ribosomes, whereas free 30S subunits and 30S subunits derived from 70S ribosomes incorporated similar amounts of 125I. Iodinated 70S ribosomes and subunits retained 62-78% of the protein synthesis activity of untreated particles and sedimentation profiles showed no gross conformational changes due to iodination. The proteins most reactive to enzymatic iodination were S4, S7, S10 and Sa of the small subunit and L2, L4, L5/9, L6 and L36 of the large subunit. Proteins S2, S3, S7, S13, Sa, L5/9, L10, L11 and L24/25 were labeled substantially more in the free subunits than in the 70S ribosome. Other proteins, including S5, S9, S12, S15/16, S18 and L36 were more extensively iodinated in the 70S ribosome than in the free subunits. The locations of tyrosine residues in some homologus ribosomal proteins from B. stearothermophilus and E. coli are compared. (orig.)

  3. Extended genetic analysis of Brazilian isolates of Bacillus cereus and Bacillus thuringiensis

    Science.gov (United States)

    Zahner, Viviane; Silva, Ana Carolina Telles de Carvalho e; de Moraes, Gabriela Pinhel; McIntosh, Douglas; de Filippis, Ivano

    2013-01-01

    Multiple locus sequence typing (MLST) was undertaken to extend the genetic characterization of 29 isolates of Bacillus cereus and Bacillus thuringiensis previously characterized in terms of presence/absence of sequences encoding virulence factors and via variable number tandem repeat (VNTR). Additional analysis involved polymerase chain reaction for the presence of sequences (be, cytK, inA, pag, lef, cya and cap), encoding putative virulence factors, not investigated in the earlier study. MLST analysis ascribed novel and unique sequence types to each of the isolates. A phylogenetic tree was constructed from a single sequence of 2,838 bp of concatenated loci sequences. The strains were not monophyletic by analysis of any specific housekeeping gene or virulence characteristic. No clear association in relation to source of isolation or to genotypic profile based on the presence or absence of putative virulence genes could be identified. Comparison of VNTR profiling with MLST data suggested a correlation between these two methods of genetic analysis. In common with the majority of previous studies, MLST was unable to provide clarification of the basis for pathogenicity among members of the B. cereus complex. Nevertheless, our application of MLST served to reinforce the notion that B. cereus and B. thuringiensis should be considered as the same species. PMID:23440117

  4. Architecture and High-Resolution Structure of Bacillus thuringiensis and Bacillus cereus Spore Coat Surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Plomp, M; Leighton, T; Wheeler, K; Malkin, A

    2005-02-18

    We have utilized atomic force microscopy (AFM) to visualize the native surface topology and ultrastructure of Bacillus thuringiensis and Bacillus cereus spores in water and in air. AFM was able to resolve the nanostructure of the exosporium and three distinctive classes of appendages. Removal of the exosporium exposed either a hexagonal honeycomb layer (B. thuringiensis) or a rodlet outer spore coat layer (B. cereus). Removal of the rodlet structure from B. cereus spores revealed an underlying honeycomb layer similar to that observed with B. thuringiensis spores. The periodicity of the rodlet structure on the outer spore coat of B. cereus was {approx}8 nm, and the length of the rodlets was limited to the cross-patched domain structure of this layer to {approx}200 nm. The lattice constant of the honeycomb structures was {approx}9 nm for both B. cereus and B. thuringiensis spores. Both honeycomb structures were composed of multiple, disoriented domains with distinct boundaries. Our results demonstrate that variations in storage and preparation procedures result in architectural changes in individual spore surfaces, which establish AFM as a useful tool for evaluation of preparation and processing ''fingerprints'' of bacterial spores. These results establish that high-resolution AFM has the capacity to reveal species-specific assembly and nanometer scale structure of spore surfaces. These species-specific spore surface structural variations are correlated with sequence divergences in a spore core structural protein SspE.

  5. Occurrence of Toxigenic Bacillus cereus and Bacillus thuringiensis in Doenjang, a Korean Fermented Soybean Paste.

    Science.gov (United States)

    Park, Kyung Min; Kim, Hyun Jung; Jeong, Moon Cheol; Koo, Minseon

    2016-04-01

    This study determined the prevalence and toxin profile of Bacillus cereus and Bacillus thuringiensis in doenjang, a fermented soybean food, made using both traditional and commercial methods. The 51 doenjang samples tested were broadly contaminated with B. cereus; in contrast, only one sample was positive for B. thuringiensis. All B. cereus isolates from doenjang were positive for diarrheal toxin genes. The frequencies of nheABC and hblACD in traditional samples were 22.7 and 0%, respectively, whereas 5.1 and 5.1% of B. cereus isolates from commercial samples possessed nheABC and hblACD, respectively. The detection rate of ces gene was 10.8%. The predominant toxin profile among isolates from enterotoxigenic B. cereus in doenjang was profile 4 (entFM-bceT-cytK). The major enterotoxin genes in emetic B. cereus were cytK, entFM, and nheA genes. The B. thuringiensis isolate was of the diarrheagenic type. These results provide a better understanding of the epidemiology of the enterotoxigenic and emetic B. cereus groups in Korean fermented soybean products.

  6. Extended genetic analysis of Brazilian isolates of Bacillus cereus and Bacillus thuringiensis

    Directory of Open Access Journals (Sweden)

    Viviane Zahner

    2013-02-01

    Full Text Available Multiple locus sequence typing (MLST was undertaken to extend the genetic characterization of 29 isolates of Bacillus cereus and Bacillus thuringiensis previously characterized in terms of presence/absence of sequences encoding virulence factors and via variable number tandem repeat (VNTR. Additional analysis involved polymerase chain reaction for the presence of sequences (be, cytK, inA, pag, lef, cya and cap, encoding putative virulence factors, not investigated in the earlier study. MLST analysis ascribed novel and unique sequence types to each of the isolates. A phylogenetic tree was constructed from a single sequence of 2,838 bp of concatenated loci sequences. The strains were not monophyletic by analysis of any specific housekeeping gene or virulence characteristic. No clear association in relation to source of isolation or to genotypic profile based on the presence or absence of putative virulence genes could be identified. Comparison of VNTR profiling with MLST data suggested a correlation between these two methods of genetic analysis. In common with the majority of previous studies, MLST was unable to provide clarification of the basis for pathogenicity among members of the B. cereus complex. Nevertheless, our application of MLST served to reinforce the notion that B. cereus and B. thuringiensis should be considered as the same species.

  7. The Bacillus anthracis Exosporium: What's the Big "Hairy" Deal?

    Science.gov (United States)

    Bozue, Joel A; Welkos, Susan; Cote, Christopher K

    2015-10-01

    In some Bacillus species, including Bacillus subtilis, the coat is the outermost layer of the spore. In others, such as the Bacillus cereus family, there is an additional layer that envelops the coat, called the exosporium. In the case of Bacillus anthracis, a series of fine hair-like projections, also referred to as a "hairy" nap, extends from the exosporium basal layer. The exact role of the exosporium in B. anthracis, or for any of the Bacillus species possessing this structure, remains unclear. However, it has been assumed that the exosporium would play some role in infection for B. anthracis, because it is the outermost structure of the spore and would make initial contact with host and immune cells during infection. Therefore, the exosporium has been a topic of great interest, and over the past decade much progress has been made to understand its composition, biosynthesis, and potential roles. Several key aspects of this spore structure, however, are still debated and remain undetermined. Although insights have been gained on the interaction of exosporium with the host during infection, the exact role and significance of this complex structure remain to be determined. Furthermore, because the exosporium is a highly antigenic structure, future strategies for the next-generation anthrax vaccine should pursue its inclusion as a component to provide protection against the spore itself during the initial stages of anthrax. PMID:26542035

  8. Bacillus as a potential diagnostic marker for yellow tongue coating.

    Science.gov (United States)

    Ye, Juan; Cai, Xueting; Yang, Jie; Sun, Xiaoyan; Hu, Chunping; Xia, Junquan; Shen, Jianping; Su, Kelei; Yan, Huaijiang; Xu, Yuehua; Zhang, Yiyan; Zhang, Sujie; Yang, Lijun; Zhi, Hao; Gao, Sizhi Paul; Yu, Qiang; Hu, Jingqing; Cao, Peng

    2016-01-01

    Observation of tongue coating, a foundation for clinical diagnosis and treatment in traditional Chinese medicine (TCM), is a major indicator of the occurrence, development, and prognosis of disease. The biological basis of tongue diagnosis and relationship between the types and microorganisms of tongue coating remain elusive. Thirteen chronic erosive gastritis (CEG) patients with typical yellow tongue coating (YTC) and ten healthy volunteers with thin white tongue coating (WTC) were included in this study. Patients were provided a 2-course targeted treatment of a herbal medicine Ban Xia Xie Xin decoction, traditionally prescribed for CEG patients with YTC, to evaluate the relationship between tongue coating microbiota and diagnosis of CEG with typical YTC. The tongue coating segregation structure was determined using Illumina Miseq sequencing of the V4-V5 region of the 16S ribosomal RNA gene. Bacillus was significantly observed only in CEG patients with YTC, but not in patients who received the decoction. YTC (n = 22) and WTC (n = 29) samples were collected for bacterial culturing to illustrate the relationship between Bacillus and YTC. The Bacillus positivity rate of YTC samples was 72.7%; Bacillus was not observed in WTC samples. In conclusion, Bacillus was strongly associated with YTC. PMID:27578261

  9. Production of Alpha Amylase by Bacillus cereus in Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Helen H. Raplong

    2014-09-01

    Full Text Available Microorganisms have the ability to secrete enzymes when they are grown in the presence of certain substrates. Amylases are among the most important industrial enzymes and are of great significance in biotechnological studies. Bacteria belonging to the genus Bacillus were isolated using mannitol egg yolk polymyxin B (MYP agar a highly selective media for Bacillus cereus isolation. The isolates were tested for α-amylase production on nutrient agar supplemented with starch and in submerged fermentation. The bacteria isolated and identified (using the Microgen Bacillus identification kit were all Bacillus cereus and SB2 had the largest zone of hydrolysis of 12mm on nutrient agar supplemented with starch as well as the highest enzyme activity of 1.62U/ml. Amylase activity of 2.56U/ml was obtained after 24 hours incubation in submerged fermentation. When amylase enzyme production parameters where optimized, maximum amylase activity was obtained at a pH of 6.5, temperature of 350C, incubation time of 24 hours and 4% inoculums concentration. Bacillus cereus SB2 is a potential isolate for alpha-amylase production with soluble starch as the sole carbon source in submerged fermentation.

  10. UJI TOKSISITAS ISOLAT Bacillus thuringiensis dari Kabupaten Lahat, Palembang, Sumatera Selatan TERHADAP LARVA NYAMUK Culex sp.

    OpenAIRE

    Chandra, Welianto

    2016-01-01

    This study aims to determine the optimal concentration of isolates of Bacillus thuringiensis to control larvae of the mosquito Culex sp. The method used is the isolation of the bacterium Bacillus thuringiensis, then the inoculation of bacteria. Bacillus thuringiensis mud samples, as much as 25 grams, obtained in the area of Lahat, South Sumatra containing Bacillus thuringiensis which includes five districts, namely Sub Gumay Talang, Jaray, Kikim West, South Kikim, and Central Kikim. Gumay ...

  11. 40 CFR 180.1128 - Bacillus subtilis MBI 600; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus subtilis MBI 600; exemption... FOOD Exemptions From Tolerances § 180.1128 Bacillus subtilis MBI 600; exemption from the requirement of... biofungicide Bacillus subtilis MBI 600 in or on all food commodities, including residues resulting from...

  12. Evaluation of in situ valine production by Bacillus subtilis in young pigs

    DEFF Research Database (Denmark)

    Nørgaard, Jan Værum; Canibe, Nuria; Assadi Soumeh, Elham;

    2016-01-01

    Mutants of Bacillus subtilis can be developed to overproduce Val in vitro. It was hypothesized that addition of Bacillus subtilis mutants to pig diets can be a strategy to supply the animal with Val. The objective was to investigate the effect of Bacillus subtilis mutants on growth performance an...

  13. Genetic Characterization of Bacillus anthracis 17 JB strain

    Directory of Open Access Journals (Sweden)

    Sakineh Seyed-Mohamadi

    2015-11-01

    Full Text Available Background and Objectives: Bacillus anthracis is one of the most homogenous bacteria ever described. Bacillus anthracis 17JB is a laboratory strain. It is broadly used as a challenge strain in guinea pigs for potency test of anthrax vaccine.Material and Methods: This work describes genetic characterization of B. anthracis 17 JB strain using the SNPs and MLVA genotyping.Results and Conclusion: In SNPs typing, the originally French 17JB strain represented the A. Br. 008/009 subgroup. In Levy's genotyping method, 843, 451 and 864 bp long fragments were identified at AA03, AJ03 and AA07 loci, respectively. In the vaccine manufacturer perspective these findings are much valuable on their own account, but similar research is required to extend molecular knowledge of B. anthracis epidemiology in Persia.Keywords: Bacillus anthracis 17JB, Genetic characterization, SNPs typing

  14. Bacillus cereus food poisoning: international and Indian perspective.

    Science.gov (United States)

    Tewari, Anita; Abdullah, Swaid

    2015-05-01

    Food borne illnesses result from eating food or drinking beverages that are contaminated with chemical matter, heavy metals, parasites, fungi, viruses and Bacteria. Bacillus cereus is one of the food-borne disease causing Bacteria. Species of Bacillus and related genera have long been troublesome to food producers on account of their resistant endospores. Their spores may be present on various types of raw and cooked foods, and their ability to survive high cooking temperatures requires that cooked foods be served hot or cooled rapidly to prevent the growth of this bacteria. Bacillus cereus is well known as a cause of food poisoning, and much more is now known about the toxins produced by various strains of this species, so that its significance in such episodes are clearer. However, it is still unclear why such cases are so rarely reported worldwide.

  15. Genotyping of Bacillus cereus strains by microarray-based resequencing.

    Directory of Open Access Journals (Sweden)

    Michael E Zwick

    Full Text Available The ability to distinguish microbial pathogens from closely related but nonpathogenic strains is key to understanding the population biology of these organisms. In this regard, Bacillus anthracis, the bacterium that causes inhalational anthrax, is of interest because it is closely related and often difficult to distinguish from other members of the B. cereus group that can cause diverse diseases. We employed custom-designed resequencing arrays (RAs based on the genome sequence of Bacillus anthracis to generate 422 kb of genomic sequence from a panel of 41 Bacillus cereus sensu lato strains. Here we show that RAs represent a "one reaction" genotyping technology with the ability to discriminate between highly similar B. anthracis isolates and more divergent strains of the B. cereus s.l. Clade 1. Our data show that RAs can be an efficient genotyping technology for pre-screening the genetic diversity of large strain collections to selected the best candidates for whole genome sequencing.

  16. BACILLUS THURINGIENSIS ELASTASES WITH INSECTICIDE ACTIVITY

    Directory of Open Access Journals (Sweden)

    E. V. Matseliukh

    2015-10-01

    Full Text Available The purpose of the research was a screening of proteases with elastase activity among Bacillus thuringiensis strains, their isolation, partially purification, study of physicochemical properties and insecticide activity in relation to the larvae of the Colorado beetle. The objects of the investigation were 18 strains of B. thuringiensis, isolated from different sources: sea water, dry biological product "Bitoksibatsillin" and also from natural populations of Colorado beetles of the Crimea, Kherson, Odesa, Mykolaiv and Zaporizhiia regions of Ukraine. Purification of enzymes with elastase activity isolated from above mentioned strains was performed by gel-chromatography and insecticide activity was studied on the 3–4 larvae instar of Colorado beetle. The ability of a number of B. thuringiensis strains to synthesize the proteases with elastase activity has been established. The most active were enzymes obtained from strains IMV B-7465, IMV B-7324 isolated from sea water, and strains 9, 902, Bt-H and 0-239 isolated from Colorado beetles. The study of the physicochemical properties of the partially purified proteases of these strains showed that they belonged to enzymes of the serine type. Peptidases of a number of B. thuringiensis strains (IMV B-7324, IMV B-7465, 902, 0-239, 9 are metal-dependent enzymes. Optimal conditions of action of all tested enzymes are the neutral and alkaline рН values and the temperatures of 30–40 °С. The studies of influence of the complex enzyme preparations and partially purified ones of B. thuringiensis strains on the larvae instar of Colorado beetles indicated that enzymes with elastase activity could be responsible for insecticide action of the tested strains.

  17. Production and Characterization of Bacillus firmus pectinase

    Directory of Open Access Journals (Sweden)

    Anna Roosdiana

    2013-03-01

    Full Text Available Pectinase is enzyme which functions to hydrolyze pectin become D-galacturonic acid unit. This enzyme is potential in various industries, especially in fruit juice industry.  Pectinase can be derived from various microorganisms resulting in different pectinase character. The aims of this research were to determine the optimum condition of pectinase production and to characterize the resulted pectinase including optimum condition of pectinase activity and the influence of metal ion.  The optimum condition of pectinase production was carried out by growing Bacillus firmus on basal media containing pectin as inducer at various  pH (5, 6, 7, 8, 9, 10, temperature (30, 35, 40, 45, 50 oC and fermentation time (6, 12, 18, 24, 30, 36 hours. while the optimum pectinase activity was done at various pH ( 4, 6, 7, 8, 10 , temperature (30, 35, 40, 45, 50 oC and reaction time (10, 20, 30, 40, 50 minutes. The influence of Zn2+, Mg2+, K+ at 2-10 mM to pectinase activity were also investigated. The result showed that optimum condition of pectinase production occurred at pH7-8, temperature 40-50 oC and fermentation time 18hours, while the optimum condition of pectinase activity was pH 7, temperature 50 oC and reaction time 30 minutes. The existence of Zn2+, Mg2+, K+ ions  affected significantly to pectinase activity.  Mg2+ acted as non competitive inhibitor; however K+ and Zn2+ acted as un competitive inhibitor.

  18. Identification and Pathogenic Potential of Clinical Bacillus and Paenibacillus Isolates.

    Science.gov (United States)

    Celandroni, Francesco; Salvetti, Sara; Gueye, Sokhna Aissatou; Mazzantini, Diletta; Lupetti, Antonella; Senesi, Sonia; Ghelardi, Emilia

    2016-01-01

    The soil-related Bacillus and Paenibacillus species have increasingly been implicated in various human diseases. Nevertheless, their identification still poses problems in the clinical microbiology laboratory and, with the exception of Bacillus anthracis and Bacillus cereus, little is known on their pathogenicity for humans. In this study, we evaluated the use of matrix-assisted laser desorption-ionization time of flight mass spectrometry (MALDI-TOF MS) in the identification of clinical isolates of these genera and conducted genotypic and phenotypic analyses to highlight specific virulence properties. Seventy-five clinical isolates were subjected to biochemical and MALDI-TOF MS identification. 16S rDNA sequencing and supplemental tests were used to solve any discrepancies or failures in the identification results. MALDI-TOF MS significantly outperformed classical biochemical testing for correct species identification and no misidentification was obtained. One third of the collected strains belonged to the B. cereus species, but also Bacillus pumilus and Bacillus subtilis were isolated at high rate. Antimicrobial susceptibility testing showed that all the B. cereus, B. licheniformis, B. simplex, B. mycoides, Paenibacillus glucanolyticus and Paenibacillus lautus isolates are resistant to penicillin. The evaluation of toxin/enzyme secretion, toxin-encoding genes, motility, and biofilm formation revealed that B. cereus displays the highest virulence potential. However, although generally considered nonpathogenic, most of the other species were shown to swim, swarm, produce biofilms, and secrete proteases that can have a role in bacterial virulence. In conclusion, MALDI-TOF MS appears useful for fast and accurate identification of Bacillus and Paenibacillus strains whose virulence properties make them of increasing clinical relevance.

  19. Identification and Pathogenic Potential of Clinical Bacillus and Paenibacillus Isolates.

    Directory of Open Access Journals (Sweden)

    Francesco Celandroni

    Full Text Available The soil-related Bacillus and Paenibacillus species have increasingly been implicated in various human diseases. Nevertheless, their identification still poses problems in the clinical microbiology laboratory and, with the exception of Bacillus anthracis and Bacillus cereus, little is known on their pathogenicity for humans. In this study, we evaluated the use of matrix-assisted laser desorption-ionization time of flight mass spectrometry (MALDI-TOF MS in the identification of clinical isolates of these genera and conducted genotypic and phenotypic analyses to highlight specific virulence properties. Seventy-five clinical isolates were subjected to biochemical and MALDI-TOF MS identification. 16S rDNA sequencing and supplemental tests were used to solve any discrepancies or failures in the identification results. MALDI-TOF MS significantly outperformed classical biochemical testing for correct species identification and no misidentification was obtained. One third of the collected strains belonged to the B. cereus species, but also Bacillus pumilus and Bacillus subtilis were isolated at high rate. Antimicrobial susceptibility testing showed that all the B. cereus, B. licheniformis, B. simplex, B. mycoides, Paenibacillus glucanolyticus and Paenibacillus lautus isolates are resistant to penicillin. The evaluation of toxin/enzyme secretion, toxin-encoding genes, motility, and biofilm formation revealed that B. cereus displays the highest virulence potential. However, although generally considered nonpathogenic, most of the other species were shown to swim, swarm, produce biofilms, and secrete proteases that can have a role in bacterial virulence. In conclusion, MALDI-TOF MS appears useful for fast and accurate identification of Bacillus and Paenibacillus strains whose virulence properties make them of increasing clinical relevance.

  20. Developments in the use of Bacillus species for industrial production.

    Science.gov (United States)

    Schallmey, Marcus; Singh, Ajay; Ward, Owen P

    2004-01-01

    Bacillus species continue to be dominant bacterial workhorses in microbial fermentations. Bacillus subtilis (natto) is the key microbial participant in the ongoing production of the soya-based traditional natto fermentation, and some Bacillus species are on the Food and Drug Administration's GRAS (generally regarded as safe) list. The capacity of selected Bacillus strains to produce and secrete large quantities (20-25 g/L) of extracellular enzymes has placed them among the most important industrial enzyme producers. The ability of different species to ferment in the acid, neutral, and alkaline pH ranges, combined with the presence of thermophiles in the genus, has lead to the development of a variety of new commercial enzyme products with the desired temperature, pH activity, and stability properties to address a variety of specific applications. Classical mutation and (or) selection techniques, together with advanced cloning and protein engineering strategies, have been exploited to develop these products. Efforts to produce and secrete high yields of foreign recombinant proteins in Bacillus hosts initially appeared to be hampered by the degradation of the products by the host proteases. Recent studies have revealed that the slow folding of heterologous proteins at the membrane-cell wall interface of Gram-positive bacteria renders them vulnerable to attack by wall-associated proteases. In addition, the presence of thiol-disulphide oxidoreductases in B. subtilis may be beneficial in the secretion of disulphide-bond-containing proteins. Such developments from our understanding of the complex protein translocation machinery of Gram-positive bacteria should allow the resolution of current secretion challenges and make Bacillus species preeminent hosts for heterologous protein production. Bacillus strains have also been developed and engineered as industrial producers of nucleotides, the vitamin riboflavin, the flavor agent ribose, and the supplement poly

  1. Sigma A recognition sites in the Bacillus subtilis genome

    DEFF Research Database (Denmark)

    Jarmer, Hanne Østergaard; Larsen, Thomas Schou; Krogh, Anders Stærmose;

    2001-01-01

    A hidden Markov model of sigma (A) RNA polymerase cofactor recognition sites in Bacillus subtilis, containing either the common or the extended -10 motifs, has been constructed based on experimentally verified sigma (A) recognition sites. This work suggests that more information exists at the ini......A hidden Markov model of sigma (A) RNA polymerase cofactor recognition sites in Bacillus subtilis, containing either the common or the extended -10 motifs, has been constructed based on experimentally verified sigma (A) recognition sites. This work suggests that more information exists...

  2. Characterization of an L-arabinose isomerase from Bacillus subtilis

    DEFF Research Database (Denmark)

    Kim, Jin-Ha; Prabhu, Ponnandy; Jeya, Marimuthu;

    2010-01-01

    An isolated gene from Bacillus subtilis str. 168 encoding a putative isomerase was proposed as an L-arabinose isomerase (L-AI), cloned into Escherichia coli, and its nucleotide sequence was determined. DNA sequence analysis revealed an open reading frame of 1,491 bp, capable of encoding a polypep......An isolated gene from Bacillus subtilis str. 168 encoding a putative isomerase was proposed as an L-arabinose isomerase (L-AI), cloned into Escherichia coli, and its nucleotide sequence was determined. DNA sequence analysis revealed an open reading frame of 1,491 bp, capable of encoding...

  3. Analysis of myo-inositol hexakisphosphate hydrolysis by Bacillus phytase

    DEFF Research Database (Denmark)

    Kerovuo, J.; Rouvinen, J.; Hatzack, Frank-Andreas

    2000-01-01

    Phytic acid (myo-inositol hexakisphosphate, InsP(6)) hydrolysis by Bacillus phytase (PhyC) was studied. The enzyme hydrolyses only three phosphates from phytic acid. Moreover, the enzyme seems to prefer the hydrolysis of every second phosphate over that of adjacent ones. Furthermore, it is very...... a reaction mechanism different from that of other phytases. By combining the data presented in this study with (1) structural information obtained from the crystal structure of Bacillus amyloliquefaciens phytase [Ha, Oh, Shin, Kim, Oh, Kim, Choi and Oh (2000) Nat. Struct. Biol. 7, 147-153], and (2) computer...

  4. Aerobic granulation of pure bacterial strain Bacillus thuringiensis

    Institute of Scientific and Technical Information of China (English)

    Sunil S ADAV; Duu-Jong LEE

    2008-01-01

    The objective of this study is to cultivate aer-obic granules by pure bacterial strain, Bacillus thuringien-sis, in a sequencing batch reactor. Stable granules sized 2.0-2.2 mm were formed in the reactor after a five-week cultivation. These granules exhibited excellent settling attributes, and degraded phenol at rates of 1.49 and concentration, respectively. Confocal laser scanning microscopic test results show that Bacillus thuringiensis was distributed over the initial small aggregates, and the outer edge of the granule was away from the core regime in the following stage.

  5. Effect of oral administration of Bacillus coagulans B37 and Bacillus pumilus B9 strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model

    Directory of Open Access Journals (Sweden)

    Lopamudra Haldar

    2016-07-01

    Full Text Available Aim: To investigate the effect of oral administration of two Bacillus strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model. Materials and Methods: An in vivo experiment was conducted for 49-day period on 36 adult male albino Wister rats divided equally into to four groups. After 7-day adaptation period, one group (T1 was fed on sterile skim milk along with basal diet for the next 28 days. Second (T2 and (T3 groups received spore biomass of Bacillus coagulans B37 and Bacillus pumilus B9, respectively, suspended in sterilized skim milk at 8-9 log colony-forming units/ml plus basal diet for 28 days, while control group (T4 was supplied with clean water along with basal diet. There was a 14-day post-treatment period. A total of 288 fecal samples (8 fecal collections per rat were collected at every 7-day interval starting from 0 to 49 days and subjected to the enumeration of the counts of coliforms and lactobacilli and Bacillus spores using respective agar media. In vitro acid and bile tolerance tests on both the strains were performed. Results: The rats those (T2 and T3 received either B. coagulans B37 or B. pumilus B9 spore along with non-fermented skim milk showed decrease (p<0.01 in fecal coliform counts and increase (p<0.05 in both fecal lactobacilli and Bacillus spore counts as compared to the control group (T4 and the group fed only skim milk (T1. In vitro study indicated that both the strains were found to survive at pH 2.0 and 3.0 even up to 3 h and tolerate bile up to 2.0% concentration even after 12 h of exposure. Conclusions: This study revealed that oral administration of either B. coagulans B37 or B. pumilus B9 strains might be useful in reducing coliform counts accompanied by concurrent increase in lactobacilli counts in the intestinal flora in rats.

  6. Ecological aspects of Bacillus thuringiensis in an Oxisol Ecologia do Bacillus thuringiensis num Latossolo

    Directory of Open Access Journals (Sweden)

    Lessandra Heck Paes Leme Ferreira

    2003-02-01

    Full Text Available Bacillus thuringiensis is a Gram positive, sporangial bacterium, known for its insecticidal habilities. Survival and conjugation ability of B. thuringiensis strains were investigated; vegetative cells were evaluated in non-sterile soil. Vegetative cells decreased rapidly in number, and after 48 hours the population was predominantly spores. No plasmid transfer was observed in non-sterile soil, probably because the cells died and the remaining cells sporulated quickly. Soil is not a favorable environment for B. thuringiensis multiplication and conjugation. The fate of purified B. thuringiensis toxin was analyzed by extractable toxin quantification using ELISA. The extractable toxin probably declined due to binding on surface-active particles in the soil.O comportamento de células vegetativas do Bacillus thuringiensis foi estudado em solo não esterilizado. Após o inóculo grande parte das células morrem e o restante esporula em 24 horas. Não foi observada conjugação provavelmente porque poucas células sobrevivem no solo e rapidamente esporulam, mostrando que este não é o ambiente propício para a multiplicação e conjugação desta bactéria. A toxina purificada, portanto livre de células, diminui rapidamente sua quantidade em solo não esterilizado. Provavelmente a ligação da toxina na fração argilosa do solo é a principal responsável por este fenômeno.

  7. Novel giant siphovirus from Bacillus anthracis features unusual genome characteristics.

    Directory of Open Access Journals (Sweden)

    Holly H Ganz

    Full Text Available Here we present vB_BanS-Tsamsa, a novel temperate phage isolated from Bacillus anthracis, the agent responsible for anthrax infections in wildlife, livestock and humans. Tsamsa phage is a giant siphovirus (order Caudovirales, featuring a long, flexible and non-contractile tail of 440 nm (not including baseplate structure and an isometric head of 82 nm in diameter. We induced Tsamsa phage in samples from two different carcass sites in Etosha National Park, Namibia. The Tsamsa phage genome is the largest sequenced Bacillus siphovirus, containing 168,876 bp and 272 ORFs. The genome features an integrase/recombinase enzyme, indicative of a temperate lifestyle. Among bacterial strains tested, the phage infected only certain members of the Bacillus cereus sensu lato group (B. anthracis, B. cereus and B. thuringiensis and exhibited moderate specificity for B. anthracis. Tsamsa lysed seven out of 25 B. cereus strains, two out of five B. thuringiensis strains and six out of seven B. anthracis strains tested. It did not lyse B. anthracis PAK-1, an atypical strain that is also resistant to both gamma phage and cherry phage. The Tsamsa endolysin features a broader lytic spectrum than the phage host range, indicating possible use of the enzyme in Bacillus biocontrol.

  8. Live-imaging of Bacillus subtilis spore germination and outgrowth

    NARCIS (Netherlands)

    R. Pandey

    2014-01-01

    Spores of Gram-positive bacteria such as Bacillus and Clostridium cause huge economic losses to the food industry. In food products, spores survive under food preservation conditions and subsequent germination and outgrowth eventually causes food spoilage. Therefore efforts are being made to elimina

  9. Global network reorganization during dynamic adaptations of Bacillus subtilis metabolism

    DEFF Research Database (Denmark)

    Buescher, Joerg Martin; Liebermeister, Wolfram; Jules, Matthieu;

    2012-01-01

    Adaptation of cells to environmental changes requires dynamic interactions between metabolic and regulatory networks, but studies typically address only one or a few layers of regulation. For nutritional shifts between two preferred carbon sources of Bacillus subtilis, we combined statistical and...

  10. The fate of Bacillus cereus in the gastrointestinal tract

    NARCIS (Netherlands)

    Pielaat A; Wijnands LM; Takumi K; Nauta MJ; Leusden FM van; MGB

    2006-01-01

    This report presents a mathematical dynamical model for the behaviour of Bacillus cereus in the gastro-intestinal tract. Biological processes and system dynamics are simultaneously incorporated in this mechanistic model. Variability in growth characteristics and physical traits of different B. cereu

  11. Transformation of undomesticated strains of Bacillus subtilis by protoplast electroporation

    NARCIS (Netherlands)

    Romero, Diego; Perez-Garcia, Alejandro; Veening, Jan-Willem; de Vicente, Antonio; Kuipers, Oscar P.; de, Vicente A.

    2006-01-01

    A rapid method combining the use of protoplasts and electroporation was developed to transform recalcitrant wild strains of Bacillus subtilis. The method described here allows transformation with both replicative and integrative plasmids, as well as with chromosomal DNA, and provides a valuable tool

  12. Antimicrobial activity of carvacrol toward Bacillus cereus on rice

    NARCIS (Netherlands)

    Ultee, A.; Slump, R.A.; Steging, G.; Smid, E.J.

    2000-01-01

    The antimicrobial activity of carvacrol, a compound present in the essential oil fraction of oreganum and thyme, toward the foodborne pathogen Bacillus cereus on rice was studied. Carvacrol showed a dose-related inhibition of growth of the pathogen. Concentrations of 0.15 mg/g and higher inhibited t

  13. Bacillus cereus: emetic toxin production and gamma hypothesis for growth

    NARCIS (Netherlands)

    Biesta-Peters, E.G.

    2011-01-01

    Bacillus cereus is a food spoilage microorganism and a pathogen. Growth of B. cereus can be prevented or delayed by adding growth limiting compounds to the food product or by altered storage conditions. Combinations of growth limiting factors

  14. Binding Affinity of Glycoconjugates to BACILLUS Spores and Toxins

    Science.gov (United States)

    Rasol, Aveen; Eassa, Souzan; Tarasenko, Olga

    2010-04-01

    Early recognition of Bacillus cereus group species is important since they can cause food-borne illnesses and deadly diseases in humans. Glycoconjugates (GCs) are carbohydrates covalently linked to non-sugar moieties including lipids, proteins or other entities. GCs are involved in recognition and signaling processes intrinsic to biochemical functions in cells. They also stimulate cell-cell adhesion and subsequent recognition and activation of receptors. We have demonstrated that GCs are involved in Bacillus cereus spore recognition. In the present study, we have investigated whether GCs possess the ability to bind and recognize B. cereus spores and Bacillus anthracis recombinant single toxins (sTX) and complex toxins (cTX). The affinity of GCs to spores + sTX and spores + cTX toxins was studied in the binding essay. Our results demonstrated that GC9 and GC10 were able to selectively bind to B. cereus spores and B. anthracis toxins. Different binding affinities for GCs were found toward Bacillus cereus spores + sTX and spores + cTX. Dilution of GCs does not impede the recognition and binding. Developed method provides a tool for simultaneous recognition and targeting of spores, bacteria toxins, and/or other entities.

  15. Molecular physiology of weak organic acid stress in Bacillus subtilis

    OpenAIRE

    Brul, S.; Beilen, van, J.W.A.

    2013-01-01

    The mechanism by which weak organic acid (WOA) preservatives inhibit growth of microorganisms may differ between different WOAs and these differences are not well understood. The aim of this thesis has been to obtain a better understanding of the mode of action of these preservatives by which they inhibit the growth of spore-forming bacteria (more specifically Bacillus subtilis).

  16. Complete Genome Sequence of Bacillus thuringiensis Bacteriophage BMBtp2

    OpenAIRE

    Dong, Zhaoxia; Peng, Donghai; Wang, Yueying; Zhu, Lei; Ruan, Lifang; Sun, Ming

    2013-01-01

    Bacillus thuringiensis is an insect pathogen which has been widely used for biocontrol. During B. thuringiensis fermentation, lysogenic bacteriophages cause severe losses of yield. Here, we announce the complete genome sequence of a bacteriophage, BMBtp2, which is induced from B. thuringiensis strain YBT-1765, which may be helpful to clarify the mechanism involved in bacteriophage contamination.

  17. Characterization of the parasporal inclusion of Bacillus thuringiensis subsp. kyushuensis.

    OpenAIRE

    Held, G. A.; Kawanishi, C. Y.; Huang, Y. S.

    1990-01-01

    Electron microscopy of Bacillus thuringiensis subsp. kyushuensis revealed that the parasporal inclusions are composed of a homogeneous center surrounded by a thick, electron-dense coating. Antibodies directed against the 135- and 65-kilodalton B. thuringiensis subsp. israelensis peptides cross-reacted with the 70- and 26-kilodalton peptides, respectively, of B. thuringiensis subsp. kyushuensis.

  18. Structural relatedness between mosquitocidal endotoxins of Bacillus thuringiensis subsp. israelensis.

    OpenAIRE

    Garduno, F; Thorne, L.; Walfield, A M; Pollock, T J

    1988-01-01

    A mosquitocidal toxin gene, cloned from Bacillus thuringiensis subsp. israelensis, was introduced into mutant crystal-negative B. thuringiensis subsp. israelensis cells. Partial toxicity to mosquitos was restored. The 58-kilodalton cloned gene product is a minor protein component of B. thuringiensis subsp. israelensis crystals and is structurally related to a major, 135-kilodalton crystal toxin.

  19. Bacillus subtilis Vegetative Catalase Is an Extracellular Enzyme

    OpenAIRE

    Naclerio, G; Baccigalupi, L; Caruso, C; De Felice, M; Ricca, E

    1995-01-01

    Strong catalase activity was secreted by Bacillus subtilis cells during stationary growth phase in rich medium but not in sporulation-inducing medium. N-terminal sequencing indicated that the secreted activity was due to the vegetative catalase KatA, previously considered an endocellular enzyme. Extracellular catalase protected B. subtilis cells from oxidative assault.

  20. Manipulating the autolytic pathway of a Bacillus protease

    NARCIS (Netherlands)

    VandenBurg, B; Eijsink, VGH; Vriend, G; Veltman, OR; Venema, G; HopsuHavu, VK; Jarvinen, M; Kirschke, H

    1997-01-01

    Autolytic degradation of Bacillus subtilis thermolysin-like proteinase (TLP-sub) is responsible for the irreversible inactivation of the enzyme at elevated temperatures. Previously, we reported five autolysis sites in B. subtilis neutral protease (Van den Burg et al., 1990, Biochem. J. 272:93-97). I

  1. Draft Genome Sequence of Biocontrol Agent Bacillus cereus UW85.

    Science.gov (United States)

    Lozano, Gabriel L; Holt, Jonathan; Ravel, Jacques; Rasko, David A; Thomas, Michael G; Handelsman, Jo

    2016-01-01

    Bacillus cereus UW85 was isolated from a root of a field-grown alfalfa plant from Arlington, WI, and identified for its ability to suppress damping off, a disease caused by Phytophthora megasperma f. sp. medicaginis on alfalfa. Here, we report the draft genome sequence of B. cereus UW85, obtained by a combination of Sanger and Illumina sequencing. PMID:27587823

  2. The transcriptionally active regions in the genome of Bacillus subtilis

    DEFF Research Database (Denmark)

    Rasmussen, Simon; Nielsen, Henrik Bjørn; Jarmer, Hanne Østergaard

    2009-01-01

    The majority of all genes have so far been identified and annotated systematically through in silico gene finding. Here we report the finding of 3662 strand-specific transcriptionally active regions (TARs) in the genome of Bacillus subtilis by the use of tiling arrays. We have measured the genome...

  3. A New Saponin Transformed from Ginsenoside Rhl by Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    Guo Hong LI; Yue Mao SHEN; Ke Qin ZHANG

    2005-01-01

    A novel saponin was isolated from the transformed products of ginsenoside Rh1 by Bacillus subtilis. It's structure was determined to be 3-O-β-D-glucopyranosyl-6-O-β-D-glucopyranosyl-20 (S)-protopanaxatriol on the basis of the spectral data.

  4. Draft Genome Sequence of Bacillus subtilis strain KATMIRA1933

    OpenAIRE

    Karlyshev, Andrey V.; Melnikov, Vyacheslav G.; Chikindas, Michael L.

    2014-01-01

    In this report, we present a draft sequence of Bacillus subtilis KATMIRA1933. Previous studies demonstrated probiotic properties of this strain partially attributed to production of an antibacterial compound, subtilosin. Comparative analysis of this strain’s genome with that of a commercial probiotic strain, B. subtilis Natto, is presented.

  5. Genome Sequence of Bacillus cereus Group Phage SalinJah.

    Science.gov (United States)

    Erill, Ivan; Caruso, Steven M

    2016-01-01

    The double-stranded DNA (dsDNA) Myoviridae Bacillus cereus group bacteriophage SalinJah was isolated from soil collected in Gyeonggi-do, South Korea. SalinJah, a cluster C phage with a broad host range, suggests the need to create a new subcluster with SalinJah and Helga as founding members. PMID:27688335

  6. The impact of manganese on biofilm development of Bacillus subtilis

    NARCIS (Netherlands)

    Mhatre, Eisha; Troszok, Agnieszka; Gallegos-Monterrosa, Ramses; Lindstädt, Stefanie; Hölscher, Theresa; Kuipers, Oscar P.; Kovács, Ákos T.

    2016-01-01

    Bacterial biofilms are dynamic and structurally complex communities, involving cell-to-cell interactions. In recent years, various environmental signals were identified that induce the complex biofilm development of the Gram-positive bacterium Bacillus subtilis. These signaling molecules are often m

  7. Thermostable, Raw-Starch-Digesting Amylase from Bacillus stearothermophilus

    OpenAIRE

    Kim, Jaeyoung; Nanmori, Takashi; Shinke, Ryu

    1989-01-01

    An endospore-forming thermophilic bacterium, which produced amylase and was identified as Bacillus stearothermophilus, was isolated from soil. The amylase had an optimum temperature of 70°C and strongly degraded wheat starch granules (93%) and potato starch granules (80%) at 60°C.

  8. Fungicidal effect of bacteriocins harvested from Bacillus spp.

    Directory of Open Access Journals (Sweden)

    Adetunji, V. O.

    2013-01-01

    Full Text Available Aims: This study investigated the ability of bacteriocins isolated from Bacillus spp. (Bacillus species to inhibit fourdifferent yeast isolates obtained from common food products (nono, yoghurt, ogi and cheese commonly consumed byNigerians with minimal heat treatment.Methodology and results: Forty-five Bacillus spp. was isolated and identified from common food products usingcultural, morphological, physiological and biochemical characteristics. These isolates were tested for antimicrobialactivity against Salmonella enteritidis (3, Micrococcus luteus (1 and Staphylococcus aureus (2. Eight bacteriocinproducing strains were identified from an over- night broth culture centrifugated at 3500 revolutions for five minutes.Fungicidal effects of these bacteriocins were tested against four yeast strains using the Agar Well Diffusion method. Thebacteriocins produced wide zones of inhibition ranging from 5.9±0.000 to 24.00±0.000 mm against the 4 yeast strainstested. There was a significant difference (at p<0.05 between the yeast organisms and the bacteriocins from theBacillus spp.Conclusion, significance and impact of study: The study reveals the antifungal property of bacteriocins from Bacillusspp. and serves therefore as a base for further studies in its use in the control of diseases and extension of shelf-life ofproducts prone to fungi contamination.

  9. Complete Genome Sequences of Nine Bacillus cereus Group Phages

    Science.gov (United States)

    Foltz, Samantha

    2016-01-01

    We report the sequences of nine novel Bacillus cereus group bacteriophages: DIGNKC, Juglone, Nemo, Nigalana, NotTheCreek, Phrodo, SageFayge, Vinny, and Zuko. These bacteriophages are double-stranded DNA-containing Myoviridae isolated from soil samples using B. thuringiensis subsp. kurstaki as the host bacterium. PMID:27417827

  10. Fatal Sepsis by Bacillus Circulans in an Immunocompromised Patient

    Directory of Open Access Journals (Sweden)

    S Jahani Sherafat

    2011-12-01

    Full Text Available An immunosuppressed man was admitted to hospital with diarrhea and a history of urinary tract infection. He was subjected to treatment with antibiotics. The patient died of putative severe sepsis. The etiological agent was a carbapenemase producing isolate of Bacillus circulans with resistance to all prescribed antimicrobial agents.

  11. Linking Bacillus cereus genotypes and carbohydrate utilization capacity

    NARCIS (Netherlands)

    Warda, Alicja K.; Siezen, Roland J.; Boekhorst, Jos; Wells-Bennik, Marjon H.J.; Jong, de Anne; Kuipers, Oscar P.; Nierop Groot, Masja N.; Abee, Tjakko

    2016-01-01

    We characterised carbohydrate utilisation of 20 newly sequenced Bacillus cereus strains isolated from food products and food processing environments and two laboratory strains, B. cereus ATCC 10987 and B. cereus ATCC 14579. Subsequently, genome sequences of these strains were analysed together wi

  12. Linking Bacillus cereus Genotypes and Carbohydrate Utilization Capacity

    NARCIS (Netherlands)

    Warda, Alicja K.; Siezen, Roland J.; Boekhorst, Jos; Wells-Bennik, Marjon H.J.; Jong, de Anne; Kuipers, Oscar P.; Nierop Groot, Masja N.; Abee, Tjakko

    2016-01-01

    We characterised carbohydrate utilisation of 20 newly sequenced Bacillus cereus strains isolated from food products and food processing environments and two laboratory strains, B. cereus ATCC 10987 and B. cereus ATCC 14579. Subsequently, genome sequences of these strains were analysed together with

  13. Induction of natural competence in Bacillus cereus ATCC14579

    NARCIS (Netherlands)

    Mironczuk, Aleksandra M.; Kovács, Ákos T.; Kuipers, O.P.

    2008-01-01

    Natural competence is the ability of certain microbes to take up exogenous DNA from the environment and integrate it in their genome. Competence development has been described for a variety of bacteria, but has so far not been shown to occur in Bacillus cereus. However, orthologues of most proteins

  14. Factors affecting the solubility of Bacillus halmapalus alpha-amylase

    DEFF Research Database (Denmark)

    Faber, Cornelius; Hobley, Timothy John; Mollerup, Jørgen;

    2008-01-01

    A detailed study of the solubility of recombinant Bacillus halmapalus alpha-amylase has been conducted. A semi-purified preparation from a bulk crystallisation was chos en that contained six isoforms with pI-values of between 5.5 and 6.1. The solubility was strongly affected by pH and could...

  15. Progress in food-related research focussing on Bacillus cereus

    NARCIS (Netherlands)

    Vries, de Y.P.; Voort, van der M.; Schaik, van W.; Hornstra, L.M.; Vos, de W.M.; Abee, T.

    2004-01-01

    Bacillus cereus is a gram-positive, rod-shaped, endospore-forming bacterium that occurs ubiquitously and is frequently isolated from soil and food products. When B. cereus is present in foods, it can cause spoilage and poisoning. The work of our group is focussed on several properties of B. cereus t

  16. Isolation and identification of local Bacillus isolates for xylanase biosynthesis.

    Directory of Open Access Journals (Sweden)

    Hassan Ammoneh

    2014-04-01

    Full Text Available Bacillus species are attractive industrial organisms due to their rapid growth rates leading to a short fermentation cycle and for their capacity to secrete important enzymes and proteins such as xylanase into the extracellular medium. Considering the industrial importance of xylanase, in this current study, Bacillus spp. were isolated from different soils and were screened for their xylanase production.Bacillus isolates used in this study were obtained from a national screening program carried out during 2006-2007 in which soil samples that covered areas throughout the interior of Syria were collected. The prepared inoculum from each of Bacillus isolates was aliquoted onto xylan agar plates, incubated at 30°C for 72 h and screened for xylanase synthesis.Xylanolytic isolates were selected depending on the clear zones of xylan hydrolysis. Fifteen isolates having the highest clearing zone were determined and grown in a solid state fermentation. Of the 15 isolates, three bacilli namely SY30A, SY185C and SY190E that showed maximum xylanase production, were identified using the 16S rDNA sequencing method. According to 16S rDNA gene sequence data, the closest phylogenetic neighbor for SY30A was Bacillus pumilus and for SY185C and SY190E isolates was Bacillus subtilis. Optimal pH and temperature for xylanase activity was 7.0 and 55ºC for SY30A and 6.0 and 60ºC for SY185C and SY190E, respectively. Under these conditions, the following activities were found to be around 1157 ± 58, 916 ± 46 and 794 ± 39 (U/g for SY30A, SY185C and SY190E, respectivly.Selected local Bacillus isolates were found to be a potential source of xylanase which was proven to be quite suitable for multiple biotechnological applications. These isolates might after extensive optimization steps be an alternative to commercially available strains.

  17. Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium Is Domain-specific.

    Science.gov (United States)

    Maes, Emmanuel; Krzewinski, Frederic; Garenaux, Estelle; Lequette, Yannick; Coddeville, Bernadette; Trivelli, Xavier; Ronse, Annette; Faille, Christine; Guerardel, Yann

    2016-04-29

    The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. It is then followed by a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization. The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. In the present study, we have explored the species and domain specificity of BclA glycosylation within the B. cereus group. First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. PMID:26921321

  18. Phylogeny and Molecular Taxonomy of the Bacillus subtilis species Complex and the Description of Bacillus subtilis subsp. inaquosorum subsp. nov

    Science.gov (United States)

    The Bacillus subtilis species complex is a tight assemblage of closely related species. For many years, it has been recognized that these species cannot be differentiated on the basis of phenotypic characteristics. Recently, it has been shown that phylogenetic analysis of the 16S ribosomal RNA gen...

  19. Comparative genomics analysis of the companion mechanisms of Bacillus thuringiensis Bc601 and Bacillus endophyticus Hbe603 in bacterial consortium.

    Science.gov (United States)

    Jia, Nan; Ding, Ming-Zhu; Gao, Feng; Yuan, Ying-Jin

    2016-01-01

    Bacillus thuringiensis and Bacillus endophyticus both act as the companion bacteria, which cooperate with Ketogulonigenium vulgare in vitamin C two-step fermentation. Two Bacillus species have different morphologies, swarming motility and 2-keto-L-gulonic acid productivities when they co-culture with K. vulgare. Here, we report the complete genome sequencing of B. thuringiensis Bc601 and eight plasmids of B. endophyticus Hbe603, and carry out the comparative genomics analysis. Consequently, B. thuringiensis Bc601, with greater ability of response to the external environment, has been found more two-component system, sporulation coat and peptidoglycan biosynthesis related proteins than B. endophyticus Hbe603, and B. endophyticus Hbe603, with greater ability of nutrients biosynthesis, has been found more alpha-galactosidase, propanoate, glutathione and inositol phosphate metabolism, and amino acid degradation related proteins than B. thuringiensis Bc601. Different ability of swarming motility, response to the external environment and nutrients biosynthesis may reflect different companion mechanisms of two Bacillus species. Comparative genomic analysis of B. endophyticus and B. thuringiensis enables us to further understand the cooperative mechanism with K. vulgare, and facilitate the optimization of bacterial consortium. PMID:27353048

  20. Comparative genomics analysis of the companion mechanisms of Bacillus thuringiensis Bc601 and Bacillus endophyticus Hbe603 in bacterial consortium.

    Science.gov (United States)

    Jia, Nan; Ding, Ming-Zhu; Gao, Feng; Yuan, Ying-Jin

    2016-06-29

    Bacillus thuringiensis and Bacillus endophyticus both act as the companion bacteria, which cooperate with Ketogulonigenium vulgare in vitamin C two-step fermentation. Two Bacillus species have different morphologies, swarming motility and 2-keto-L-gulonic acid productivities when they co-culture with K. vulgare. Here, we report the complete genome sequencing of B. thuringiensis Bc601 and eight plasmids of B. endophyticus Hbe603, and carry out the comparative genomics analysis. Consequently, B. thuringiensis Bc601, with greater ability of response to the external environment, has been found more two-component system, sporulation coat and peptidoglycan biosynthesis related proteins than B. endophyticus Hbe603, and B. endophyticus Hbe603, with greater ability of nutrients biosynthesis, has been found more alpha-galactosidase, propanoate, glutathione and inositol phosphate metabolism, and amino acid degradation related proteins than B. thuringiensis Bc601. Different ability of swarming motility, response to the external environment and nutrients biosynthesis may reflect different companion mechanisms of two Bacillus species. Comparative genomic analysis of B. endophyticus and B. thuringiensis enables us to further understand the cooperative mechanism with K. vulgare, and facilitate the optimization of bacterial consortium.

  1. Bacillus thuringiensis: legado para el siglo XXI Bacillus thuringiensis: the legacy to the XXI century

    Directory of Open Access Journals (Sweden)

    Orduz S.

    1998-06-01

    Full Text Available

    Los insecticidas basados en la bacteria Bacillus thuringiensis son el principal renglón productivo del mercado mundial de biopesticidas. La investigación dedicada a esta área, promovida por la urgente necesidad de resolver problemas agrícolas y de salud pública, ha dado lugar a un conocimiento exhaustivo de su biología. La diversidad de cepas diferentes de B. thuringiensis ha permitido desarrollar productos principalmente, pero no exclusivamente, para el control de insectos. Con los nuevos desarrollos de la biología molecular, se ha logrado comprender su mecanismo de acción a nivel molecular y también se ha logrado extender sus capacidades entomopatógenas. Como producto de su amplio uso en muchos países, se han presentado casos de resistencia en poblaciones de insectos susceptibles. Con esta revisión se pretende elaborar un contexto teórico del estado actual de la investigación sobre B. thuringiensis, describiendo brevemente el conocimiento sobre esta bacteria, haciendo hincapié en los fenómenos biológicos que subyacen su actividad tóxica y la problemática que se avecina en el próximo siglo con los fenómenos de resistencia cada vez más comunes, todo esto analizado desde una perspectiva biotecnológica.

    Bacillus thuringiensis-based insecticides are the main production line of the biopesticides world market. The research devoted to this area, promoted by the necessity to solve problems in agriculture and public health has resulted in an exhaustive knowledge of its biology. The diversity of the B. thuringiensis strains has permitted to develop several products mainly, but not exclusively, for insect control. With the new developments in the field of molecular biology, it has been possible to understand the molecular basis of the mode of action and to increase the range of activity as well. As a result

  2. Isolation and characterization of protease from Bacillus subtilis 1012M15

    Directory of Open Access Journals (Sweden)

    ELFI SUSANTI

    2003-01-01

    Full Text Available A local strain of Bacillus sp. BAC4, is known to produce penicillin G acylase (PGA enzyme with relatively high activity. This strain secretes the PGA into the culture medium. However, it has been reported that PGA activity fall and rise during culture, and the activity plummets during storege at –200C, which probably due to usage protease activity of Bacillus sp. BAC4. To study the possible use of Bacillus subtilis 1012M15 as a host cell for cloning the pga gene from Bacillus sp. BAC4, the protease activity of Bacillus subtilis 1012M15 were studied. Protease activity was determined by Horikoshi method. In this experiment, maximum protease activity in Bacillus subtilis 1012M15 culture was obsereved after 8 hours. At this optimum condition, protease activity of Bacillus sp. BAC4 is five time higher than that of Bacillus subtilis 1012M15. This situation promised the possible usage of Bacillus subtilis 1012M15 as a host cell for pga expression. For protease characterization, the bacterial culture had been separated from the cell debris by centrifugation. The filtrate was concentrated by freeze drying, fractionated by ammonium sulphate, dialyzed in selovan tube, and then fractionated by ion exchance chromatography employing DEAE-cellulose. The five peaks resulted indicated the presence of five protease. Based on inhibitor and activator influence analysis, it could be concluded that proteases from Bacillus subtilis 1012M15 contained of serin protease as well as metalloprotease and serin protease mixture.

  3. Characterization and functional expression in Escherichia coli of the sodium/proton/glutamate symport proteins of Bacillus stearothermophilus and Bacillus caldotenax

    NARCIS (Netherlands)

    TOLNER, B; POOLMAN, B; KONINGS, WN

    1992-01-01

    The genes encoding the Na+/H+/L-glutamate symport proteins of the thermophilic organisms Bacillus stearothermophilus (gltT(Bs)) and Bacillus caldotenax (gltT(Bc)) were cloned by complementation of Escherichia coli JC5412 for growth on glutamate as sole source of carbon, energy and nitrogen. The nucl

  4. Genetic analysis of Bacillus stearothermophilus by protoplast fusion

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Z.; Wojcik, S.F.; Welker, N.E.

    1986-03-01

    Efficient and reliable protoplasting, regeneration, and fusion techniques were established for the prototrophic strain Bacillus stearothermophilus NUB36. Auxotrophic mutants were isolated, and protoplast fusion was used to construct isogenic mutant strains and for chromosomal mapping. Markers were mapped using two-, three-, and four-factor crosses. The order of the markers was hom-1-thr-1-his-1-(gly-1 or gly-2)-pur-1-pur-2. These markers may be analogous to hom, thrA, hisA, glyC, and purA markers on the Bacillus subtilis chromosome. No analogous pur-1 marker has been reported in B. subtilis. The relative order of three of the markers (hom-1-thr-1-gly-1) was independently confirmed by transduction.

  5. The Bacillus cereus spoIIS programmed cell death system

    Directory of Open Access Journals (Sweden)

    Jana eMelnicakova

    2015-08-01

    Full Text Available Programmed cell death in bacteria is generally associated with two¬ component toxin antitoxin systems. The SpoIIS toxin-antitoxin system, consisting of a membrane bound SpoIISA toxin and a small, cytosolic antitoxin SpoIISB, was originally identified in Bacillus subtilis. In this work we describe the Bacillus cereus SpoIIS system which is a three-component system, harbouring an additional gene spoIISC. Its protein product serves as an antitoxin, and similarly as SpoIISB, is able to bind SpoIISA and abolish its toxic effect. Our results indicate that SpoIISC seems to be present not only in B. cereus but also in other Bacilli containing a SpoIIS toxin antitoxin system. In addition, we show that B. cereus SpoIISA can form higher oligomers and we discuss the possible role of this multimerization for the protein’s toxic function.

  6. MOLECULAR PROFILING AND ANTIMICROBIAL ACTIVITY OF BACTERIOCIN FROM BACILLUS SUBTILIS

    Directory of Open Access Journals (Sweden)

    Berlina Dhas S

    2012-12-01

    Full Text Available Development of multi drug resistant organism has been high due to improper use of antibiotics. That made the necessity to develop new drug molecules. In this study an effort was made to find a new alternative. A wild type microorganism was isolated from soil and was identified as Bacillus and confirmed as Bacillus subtilis species by 16S r RNA sequencing. The strain was identified to have the ability to produce bacteriocin by stab overlay assay. Bacteriocin was produced in nutrient broth and that was extracted by organic solvent extraction using chloroform and further purification was carried out by HPLC and the molecular weight of the bacteriocin was analysed by SDSPAGE. Antimicrobial activity was analysed on four strains Pseudomonas sp, Staphylococcus sp, Klebsiella sp and Proteus sp. and was found to be sensitive towards the analyzed strains.

  7. ANOTHER NITROGEN-FIXING MICROORGANISM IN SUGARCANE STALKS: Bacillus brevis?

    Directory of Open Access Journals (Sweden)

    Lorelí de los A. Mirabal

    2000-01-01

    Full Text Available Con el objetivo de identificar un microorganismo formador de colonias blancas en medio de cultivo LGI, prove- niente de savia apoplástica del tallo de la caña de azúcar, se realizaron diferentes experimentos en los que se utilizó la variedad ML-318. De la misma se extrajo savia apoplástica, de la cual se aisló el microorganismo de interés, al que se le realizaron pruebas morfológicas, culturales y bioquímicas, las que revelaron un 65 % de probabilidad de que el microorganismo de interés se corresponde con Bacillus brevis. Se detectó actividad nitrogenasa por la reducción de acetileno a etileno. Además, los resultados indican interacción entre Acetobacter diazotrophicus y el microorganismo formador de colonias blancas identificado como Bacillus brevis.

  8. Production of Protocatechuic Acid in Bacillus Thuringiensis ATCC33679

    Directory of Open Access Journals (Sweden)

    Bianca L. Garner

    2012-03-01

    Full Text Available Protocatechuic acid, or 3,4-dihydroxybenzoic acid, is produced by both soil and marine bacteria in the free form and as the iron binding component of the siderophore petrobactin. The soil bacterium, Bacillus thuringiensis kurstaki ATCC 33679, contains the asb operon, but does not produce petrobactin. Iron restriction resulted in diminished B. thuringiensis kurstaki ATCC 33679 growth and the production of catechol(s. The gene product responsible for protocatechuic acid (asbF and its receptor (fatB were expressed during stationary phase growth. Gene expression varied with growth temperature, with optimum levels occurring well below the Bacillus anthracis virulence temperature of 37 °C. Regulation of protocatechuic acid suggests a possible role for this compound during soil growth cycles.

  9. Biodegradation of furfural by Bacillus subtilis strain DS3.

    Science.gov (United States)

    Zheng, Dan; Bao, Jianguo; Lu, Jueming; Lv, Quanxi

    2015-07-01

    An aerobic bacterial strain DS3, capable of growing on furfural as sole carbon source, was isolated from actived sludge of wastewater treatment plant in a diosgenin factory after enrichment. Based on morphological physiological tests as well as 16SrDNA sequence and Biolog analyses it was identified as Bacillus subtilis. The study revealed that strain DS3 utilized furfural, as analyzed by high-performance liquid chromatography (HPLC). Under following conditions: pH 8.0, temperature 35 degrees C, 150 rpm and 10% inoculum, strain DS3 showed 31.2% furfural degradation. Furthermore, DS3 strain was found to tolerate furfural concentration as high as 6000 mg(-1). The ability of Bacillus subtilis strain DS3 to degrade furfural has been demonstrated for the first time in the present study.

  10. Novel routes for improving biocontrol activity of Bacillus based bioinoculants

    Directory of Open Access Journals (Sweden)

    Liming eWu

    2015-12-01

    Full Text Available Biocontrol formulations prepared from plant-growth-promoting bacteria are increasingly applied in sustainable agriculture. Especially inoculants prepared from endospore-forming Bacillus strains have been proven as efficient and environmental-friendly alternative to chemical pesticides due to their long shelf life, which is comparable with that of agrochemicals. However, these formulations of the first generation are sometimes hampered in their action and do not fulfill in each case the expectations of the appliers. In this review we use the well-known plant-associated Bacillus amyloliquefaciens type strain FZB42 as example for the successful application of different techniques offered today by comparative, evolutionary and functional genomics, site-directed mutagenesis and strain construction including marker removal, for paving the way for preparing a novel generation of biocontrol agents.

  11. Optimization Conditions of Production Fibrinolytic Enzyme from Bacillus lichniformis B4 Local Isolate

    OpenAIRE

    Essam F. Al-Juamily; Bushra H. Al-Zaidy

    2012-01-01

    The study was conducted with the aim to found local isolate belongs to Bacillus lichniformis to produce fibrinolytic enzyme with highest activity under optimal conditions. Forty-five local isolates belongs to the genus Bacillus lichniformis were selected for production of fibrinolytic enzyme (E.C. 3.4.). The isolate Bacillus lichniformis B4 was selected due to its high productivity of fibrinolytic enzyme. The optimal conditions for fibrinolytic enzyme production were determined, using a solid...

  12. Pengaruh Sumber Karbon dan Nitrogen Terhadap Pertumbuhan dan Pembentukan Spora Bacillus sp. Bk17

    OpenAIRE

    Rachmi

    2014-01-01

    Study of the effect of carbon and nitrogen sources on the growth and sporulation of Bacillus sp. BK17 has been conducted. Bacillus sp. BK17 was cultured in broth medium containing different carbon and nitrogen sources, and incubated for 6 days. The highest growth of Bacillus sp. BK17 was obtained on molasses-tryptone medium incubated during 3 days and the lowest growth obtained on whey-sodium nitrate. The highest spore formation was also obtained from molasses-tryptone mediu...

  13. Bacillus anthracis infections – new possibilities of treatment

    OpenAIRE

    Dorota Żakowska; Michał Bartoszcze; Marcin Niemcewicz; Agata Bielawska-Drózd; Józef Knap; Piotr Cieślik; Krzysztof Chomiczewski; Janusz Kocik

    2015-01-01

    [b]Introduction and objective[/b]. [i]Bacillus anthracis[/i] is one of biological agents which may be used in bioterrorism attacks. The aim of this study a review of the new treatment possibilities of anthrax, with particular emphasis on the treatment of pulmonary anthrax. [b]Abbreviated description of the state of knowledge[/b]. Pulmonary anthrax, as the most dangerous clinical form of the disease, is also extremely difficult to treat. Recently, considerable progress in finding new dru...

  14. Immunological analysis of cell-associated antigens of Bacillus anthracis.

    OpenAIRE

    Ezzell, J W; Abshire, T. G.

    1988-01-01

    Sera from Hartley guinea pigs vaccinated with a veterinary live spore anthrax vaccine were compared with sera from guinea pigs vaccinated with the human anthrax vaccine, which consists of aluminum hydroxide-adsorbed culture proteins of Bacillus anthracis V770-NP-1R. Sera from animals vaccinated with the spore vaccine recognized two major B. anthracis vegetative cell-associated proteins that were either not recognized or poorly recognized by sera from animals that received the human vaccine. T...

  15. Resistance of Bacillus amyloliquefaciens spores to melt extrusion process conditions

    OpenAIRE

    Ciera, Lucy Wanjiru; Beladjal, Lynda; Almeras, Xavier; Gheysens, Tom; Nierstrasz, Vincent; Van Langenhove, Lieva; Mertens, Johan

    2014-01-01

    With the increasing demand for functionalised textile materials, industry is focusing on research that will add novel properties to textiles. Bioactive compounds and their benefits have been and are still considered as a possible source of unique functionalities to be explored. However, incorporating bioactive compounds into textiles and their resistance to textile process parameters has not yet been studied. In this study, we developed a system to study the resistance of Bacillus amyloliquef...

  16. Identification of Bacillus Strains for Biological Control of Catfish Pathogens

    OpenAIRE

    Ran, Chao; Carrias, Abel; Williams, Malachi A.; Capps, Nancy; Dan, Bui C. T.; Newton, Joseph C.; Joseph W Kloepper; Ooi, Ei L.; Browdy, Craig L.; Terhune, Jeffery S.; Liles, Mark R.

    2012-01-01

    Bacillus strains isolated from soil or channel catfish intestine were screened for their antagonism against Edwardsiella ictaluri and Aeromonas hydrophila, the causative agents of enteric septicemia of catfish (ESC) and motile aeromonad septicaemia (MAS), respectively. Twenty one strains were selected and their antagonistic activity against other aquatic pathogens was also tested. Each of the top 21 strains expressed antagonistic activity against multiple aquatic bacterial pathogens including...

  17. Respiratory systems of the Bacillus cereus mother cell and forespore.

    OpenAIRE

    Escamilla, J E; R. Ramírez; Del-Arenal, P; Aranda, A.

    1986-01-01

    The respiratory systems of the mother cells and forespores of Bacillus cereus were compared throughout the maturation stages (III to VI) of sporulation. The results indicated that both cell compartments contain the same assortment of oxidoreductases and cytochromes. However membrane fractions from young forespores were clearly distinct from those of the mother cell, i.e., lower content of cytochrome aa3, lower cytochrome c oxidase activity, higher concentration of cytochrome o, and a lower se...

  18. Biodegradation of Eugenol by Bacillus Cereus Strain PN24

    OpenAIRE

    Kadakol, Jagannath C.; Kamanavalli, Chandrappa M.

    2010-01-01

    Bacillus cereus PN24 was isolated from soil by a conventional enrichment culture method using eugenol as a sole source of carbon and energy. The organism also utilized eugenol, 4-vinyl guaiacol, vanillin, vanillic acid and protocatechuic acid as growth substrates. The organism degraded eugenol to protocatechuic acid, which was further metabolized by a β-ketoadipate pathway. On the other hand, the intermediate of the eugenol-degrading pathway, such as ferulic acid was not detected in the cultu...

  19. Water surface tension modulates the swarming mechanics of Bacillus subtilis

    OpenAIRE

    Ke, Wan-Ju; Hsueh, Yi-Huang; Cheng, Yu-Chieh; Wu, Chih-Ching; Liu, Shih-Tung

    2015-01-01

    Many Bacillus subtilis strains swarm, often forming colonies with tendrils on agar medium. It is known that B. subtilis swarming requires flagella and a biosurfactant, surfactin. In this study, we find that water surface tension plays a role in swarming dynamics. B. subtilis colonies were found to contain water, and when a low amount of surfactin is produced, the water surface tension of the colony restricts expansion, causing bacterial density to rise. The increased density induces a quorum ...

  20. Mechanism of Insect Resistance to the Microbial Insecticide Bacillus thuringiensis

    Science.gov (United States)

    van Rie, J.; McGaughey, W. H.; Johnson, D. E.; Barnett, B. D.; van Mellaert, H.

    1990-01-01

    Receptor binding studies show that resistance of a laboratory-selected Plodia interpunctella strain to a Bacillus thuringiensis insecticidal crystal protein (ICP) is correlated with a 50-fold reduction in affinity of the membrane receptor for this protein. The strain is sensitive to a second type of ICP that apparently recognizes a different receptor. Understanding the mechanism of resistance will provide strategies to prevent or delay resistance and hence prolong the usefulness of B. thuringiensis ICPs as environmentally safe insecticides.

  1. Live-imaging of Bacillus subtilis spore germination and outgrowth

    OpenAIRE

    Pandey, R

    2014-01-01

    Spores of Gram-positive bacteria such as Bacillus and Clostridium cause huge economic losses to the food industry. In food products, spores survive under food preservation conditions and subsequent germination and outgrowth eventually causes food spoilage. Therefore efforts are being made to eliminate or inactivate these bacterial spores in foods. In this regard food industry uses different preservation methods such as thermal-treatment, weak acids, antimicrobial compounds etc. Complete therm...

  2. Carbohydrate metabolism in the mosquito pathogen Bacillus sphaericus 2362.

    OpenAIRE

    Russell, B L; Jelley, S A; Yousten, A A

    1989-01-01

    Bacillus sphaericus 2362 is pathogenic for mosquito larvae and is being considered for large-scale production as a larvicide. The inability of the bacteria to metabolize carbohydrates requires that they be grown on proteinaceous media. This bacterium was found to be unable to transport glucose or sucrose into the cell, and it lacked glucokinase and hexokinase activity. In addition, it lacked phosphoglucose isomerase, phosphofructokinase, and glucose 6-phosphate dehydrogenase, which are early ...

  3. Monarch larvae sensitivity to Bacillus thuringiensis- purified proteins and pollen

    OpenAIRE

    Hellmich, Richard L; Blair D Siegfried; Sears, Mark K.; Stanley-Horn, Diane E.; Daniels, Michael J.; Mattila, Heather R.; Spencer, Terrence; Bidne, Keith G.; Lewis, Leslie C.

    2001-01-01

    Laboratory tests were conducted to establish the relative toxicity of Bacillus thuringiensis (Bt) toxins and pollen from Bt corn to monarch larvae. Toxins tested included Cry1Ab, Cry1Ac, Cry9C, and Cry1F. Three methods were used: (i) purified toxins incorporated into artificial diet, (ii) pollen collected from Bt corn hybrids applied directly to milkweed leaf discs, and (iii) Bt pollen contaminated with corn tassel material applied directly to milkweed leaf discs. Bioassays of purified Bt tox...

  4. The Phylloplane as a Source of Bacillus thuringiensis Variants

    OpenAIRE

    Smith, Robert A.; Couche, Graham A.

    1991-01-01

    Novel variants of Bacillus thuringiensis were isolated from the phylloplane of deciduous and conifer trees as well as of other plants. These isolates displayed a range of toxicity towards Trichoplusia ni. Immunoblot and toxin protein analysis indicate that these strains included representatives of the three principal B. thuringiensis pathotypes active against larvae of the orders Lepidoptera, Diptera, and Coleoptera. We propose that B. thuringiensis be considered part of the common leaf micro...

  5. Worldwide Abundance and Distribution of Bacillus thuringiensis Isolates

    OpenAIRE

    Martin, Phyllis A. W.; Travers, Russell S.

    1989-01-01

    We found the insect control agent Bacillus thuringiensis to be a ubiquitous soil microorganism. Using acetate selection to screen soil samples, we isolated B. thuringiensis in 785 of 1,115 soil samples. These samples were obtained in the United States and 29 other countries. A total of 48% of the B. thuringiensis isolates (8,916 isolates) fit the biochemical description of known varieties, while 52% represented undescribed B. thuringiensis types. Over 60% (1,052 isolates) of the isolates test...

  6. Functional Comparison of the Two Bacillus anthracis Glutamate Racemases▿

    OpenAIRE

    Dodd, Dylan; Reese, Joseph G.; Louer, Craig R.; Ballard, Jimmy D.; Spies, M. Ashley; Blanke, Steven R.

    2007-01-01

    Glutamate racemase activity in Bacillus anthracis is of significant interest with respect to chemotherapeutic drug design, because l-glutamate stereoisomerization to d-glutamate is predicted to be closely associated with peptidoglycan and capsule biosynthesis, which are important for growth and virulence, respectively. In contrast to most bacteria, which harbor a single glutamate racemase gene, the genomic sequence of B. anthracis predicts two genes encoding glutamate racemases, racE1 and rac...

  7. The Pore-Forming Haemolysins of Bacillus Cereus: A Review

    OpenAIRE

    Vincent Sanchis; Nalini Ramarao

    2013-01-01

    The Bacillus cereus sensu lato group contains diverse Gram-positive spore-forming bacteria that can cause gastrointestinal diseases and severe eye infections in humans. They have also been incriminated in a multitude of other severe, and frequently fatal, clinical infections, such as osteomyelitis, septicaemia, pneumonia, liver abscess and meningitis, particularly in immuno-compromised patients and preterm neonates. The pathogenic properties of this organism are mediated by the synergistic ef...

  8. [Depolymerization of chitosan by chinolytic complex from Bacillus sp. 739].

    Science.gov (United States)

    Il'ina, A V; Varlamov, V P; Melent'ev, A I; Aktuganov, G E

    2001-01-01

    Low-molecular-weight (3-6 kDa) water-soluble chitosan was obtained by enzymatic depolymerization. Hydrolysis of crab chitosan was induced by O-glycoside hydrolase (EC 3.2.1), an extracellular chitinolytic complex from Bacillus sp. 739. The optimum conditions for hydrolysis were found (sodium-acetate buffer, pH 5.2; 55 degrees C; an enzyme/substrate ratio 4 U/g chitosan; 1 h).

  9. Defensive strategies of Bacillus anthracis that promote a fatal disease

    OpenAIRE

    Mogridge, Jeremy

    2007-01-01

    Bacillus anthracis is a Gram-positive bacterium that causes anthrax. Bacterial spores that enter the host germinate into metabolically active bacilli that disseminate throughout the body and replicate to high numbers. Two virulence factors are essential for this unrestrained growth. The first is a weakly immunogenic poly γ-D-glutamic acid capsule that surrounds the bacilli and confers resistance to phagocytosis. The second virulence factor, anthrax toxin, disrupts multiple host functions to d...

  10. Molecular Epidemiology of Bacillus anthracis: Determining the Correct Origin▿

    OpenAIRE

    Pilo, Paola; Perreten, Vincent; Frey, Joachim

    2008-01-01

    We analyzed and compared strains of Bacillus anthracis isolated from husbandry and industrial anthrax cases in Switzerland between 1952 and 1981 with published data using multiple-locus variable-number tandem repeat analysis. Strains isolated from autochthonous cases of anthrax in cattle belong to genotype B2, together with strains from continental Europe, while human B. anthracis strains clustered with genotype A4. These strains could be traced back to outbreaks of human anthrax that occurre...

  11. Cytokine Response to Infection with Bacillus anthracis Spores

    OpenAIRE

    Pickering, Alison K.; Osorio, Manuel; Lee, Gloria M.; Grippe, Vanessa K.; Bray, Mechelle; Merkel, Tod J.

    2004-01-01

    Bacillus anthracis, the etiological agent of anthrax, is a gram-positive, spore-forming bacterium. The inhalational form of anthrax is the most severe and is associated with rapid progression of the disease and the outcome is frequently fatal. Transfer from the respiratory epithelium to regional lymph nodes appears to be an essential early step in the establishment of infection. This transfer is believed to occur by means of carriage within alveolar macrophages following phagocytosis. Therefo...

  12. Historical Distribution and Molecular Diversity of Bacillus anthracis, Kazakhstan

    OpenAIRE

    Aikembayev, Alim M.; Lukhnova, Larissa; Temiraliyeva, Gulnara; Meka-Mechenko, Tatyana; Pazylov, Yerlan; Zakaryan, Sarkis; Denissov, Georgiy; Easterday, W. Ryan; Matthew N. Van Ert; Keim, Paul; Francesconi, Stephen C.; Jason K Blackburn; Hugh-Jones, Martin; Hadfield, Ted

    2010-01-01

    To map the distribution of anthrax outbreaks and strain subtypes in Kazakhstan during 1937–2005, we combined geographic information system technology and genetic analysis by using archived cultures and data. Biochemical and genetic tests confirmed the identity of 93 archived cultures in the Kazakhstan National Culture Collection as Bacillus anthracis. Multilocus variable number tandem repeat analysis genotyping identified 12 genotypes. Cluster analysis comparing these genotypes with previousl...

  13. Production and purification of Bacillus anthracis protective antigen

    OpenAIRE

    2005-01-01

    Protective antigen (PA) plays crucial roles in the pathogenicity and virulence of Bacillus anthracis. Animals or human immunised with the protein acquire a complete protection against the disease. In addition to vaccine, PA can also be developed into a sensitive diagnostic test for anthrax. The purpose of this study was to produce PA using a culture medium easily obtained, and to develop a simple and effective technique for purification of the protein. To produce PA, B. anthracis Sterne 34F2 ...

  14. Bacillus anthracis IsdG, a Heme-Degrading Monooxygenase

    OpenAIRE

    Skaar, Eric P.; Gaspar, Andrew H.; Schneewind, Olaf

    2006-01-01

    Bacillus anthracis, the causative agent of anthrax, utilizes hemin and hemoglobin for growth in culture, suggesting that these host molecules serve as sources for the nutrient iron during bacterial infection. Bioinformatic analyses of the B. anthracis genome revealed genes with similarity to the iron-regulated surface determinant (isd) system responsible for heme uptake in Staphylococcus aureus. We show that the protein product of one of these genes, isdG, binds hemin in a manner resembling t...

  15. Glycerol Monolaurate Inhibits Virulence Factor Production in Bacillus anthracis

    OpenAIRE

    Vetter, Sara M; Schlievert, Patrick M.

    2005-01-01

    Anthrax, caused by Bacillus anthracis, has been brought to the public's attention because of the 2001 bioterrorism attacks. However, anthrax is a disease that poses agricultural threats in the United States as well as human populations in Europe, China, Africa, and Australia. Glycerol monolaurate (GML) is a compound that has been shown to inhibit exotoxin production by Staphylococcus aureus and other gram-positive bacteria. Here, we study the effects of GML on growth and toxin production in B...

  16. Biosynthetic Analysis of the Petrobactin Siderophore Pathway from Bacillus anthracis▿

    OpenAIRE

    Lee, Jung Yeop; Janes, Brian K.; Passalacqua, Karla D; Pfleger, Brian F.; Bergman, Nicholas H; Liu, Haichuan; Håkansson, Kristina; Somu, Ravindranadh V.; Aldrich, Courtney C.; Cendrowski, Stephen; Hanna, Philip C.; Sherman, David H.

    2006-01-01

    The asbABCDEF gene cluster from Bacillus anthracis is responsible for biosynthesis of petrobactin, a catecholate siderophore that functions in both iron acquisition and virulence in a murine model of anthrax. We initiated studies to determine the biosynthetic details of petrobactin assembly based on mutational analysis of the asb operon, identification of accumulated intermediates, and addition of exogenous siderophores to asb mutant strains. As a starting point, in-frame deletions of each of...

  17. Biocontrol: Bacillus penetrans and Related Parasites of Nematodes

    OpenAIRE

    Sayre, R. M.

    1980-01-01

    Bacillus penetrans Mankau, 1975, previously described as Duboscqia penetrans Thorne 1940, is a candidate agent for biocontrol of nematodes. This review considers the life stages of this bacterium: vegetative growth phase, colony fragmentation, sporogenesis, soil phase, spore attachment, and penetration into larvae of root-knot nematodes. The morphology of the microthallus colonies and the unusual external features of the spore are discussed. Taxonomic affinities with the actinomycetes, partic...

  18. The Regulation of Exosporium-Related Genes in Bacillus thuringiensis

    Science.gov (United States)

    Peng, Qi; Kao, Guiwei; Qu, Ning; Zhang, Jie; Li, Jie; Song, Fuping

    2016-01-01

    Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis (Bt) are spore-forming members of the Bacillus cereus group. Spores of B. cereus group species are encircled by exosporium, which is composed of an external hair-like nap and a paracrystalline basal layer. Despite the extensive studies on the structure of the exosporium-related proteins, little is known about the transcription and regulation of exosporium gene expression in the B. cereus group. Herein, we studied the regulation of several exosporium-related genes in Bt. A SigK consensus sequence is present upstream of genes encoding hair-like nap proteins (bclA and bclB), basal layer proteins (bxpA, bxpB, cotB, and exsY ), and inosine hydrolase (iunH). Mutation of sigK decreased the transcriptional activities of all these genes, indicating that the transcription of these genes is controlled by SigK. Furthermore, mutation of gerE decreased the transcriptional activities of bclB, bxpB, cotB, and iunH but increased the expression of bxpA, and GerE binds to the promoters of bclB, bxpB, cotB, bxpA, and iunH. These results suggest that GerE directly regulates the transcription of these genes, increasing the expression of bclB, bxpB, cotB, and iunH and decreasing that of bxpA. These findings provide insight into the exosporium assembly process at the transcriptional level. PMID:26805020

  19. Cytolytic Toxin and Related Genes in Bacillus thuringiensis

    Institute of Scientific and Technical Information of China (English)

    QI Dong-lai; LI Yi-dan; GAO Ji-guo

    2005-01-01

    Bacillus thuringiensis is a ubiquitous gram-positive, spore-forming bacterium that forms parasporal crystal during the stationary phase of its growth cycle. These crystal proteins, including Cry and Cyt protein, are toxic to certain insects. Lately, some problems about Cyt classification, structural characteristic, action mechanism and resistance to Cyt toxin are becoming new hotspots. We review the progress of above problems in several foreign labs.

  20. Expression of alpha-amylase in Bacillus licheniformis.

    OpenAIRE

    Rothstein, D. M.; Devlin, P E; Cate, R. L.

    1986-01-01

    In Bacillus licheniformis, alpha-amylase production varied more than 100-fold depending on the presence or absence of a catabolite-repressing carbon source in the growth medium. alpha-Amylase was produced during the growth phase and not at the onset of the stationary phase. Induction of alpha-amylase correlated with synthesis of mRNA initiating at the promoter of the alpha-amylase gene.

  1. Studies on the fermentation of bacillus thuringiensis var israelensis

    OpenAIRE

    Pearson, Dermot

    1985-01-01

    During this work the fermentation of Bacillus thuringiensis var israelensis under industrial conditions was studied with respect to the development of a process for the production of a mosquitocidal insecticide elaborated by this organism. This was done by the development of a two-stage inoculum protocol which produced a high biomass-containing inoculum of vegetative cells which were found to be preferable to free spores for use as an inoculum source. In order to optimize the production st...

  2. Characterization of Bacillus amyloliquefacien contaminating 75% alcohol disinfectant

    Institute of Scientific and Technical Information of China (English)

    Wanming ZHANG; Yuesha YUAN; Cangli BIAN; Wen ZHANG; Lan WANG; Xianyu TU; Huqiang HUANG

    2008-01-01

    The clinical characterization of Bacillus amy-loliquefacien contaminating 75% alcohol disinfectants were studied. The bacteria were cultured and observed by using bacterial examination under the hospital infec-tion monitor. According to the regulations, the resistance of bacterial to physical and chemical factors was tested. Drug sensitivity tests for 20 commonly used medicines were carried out using a K-B method. The bacterial plas-mids were analyzed using the Birnboim method. The bac-teria were found after being cultured in the clinically-used 75% alcohol disinfectant fluid. Their total number was more than 800 cfu/mL and they were identified as Bacillus amyloliquefacien. The bacteria were also found to be resistant to boiling for 5 min. It grew well in 95% alcohol disinfectant and was insensitive to 84 disinfectant fluids containing chlorine (1000 mg/L) and such disinfec-tants as ozone. They were able to be sterilized better through routine ultraviolet exposure for 30 min or gas pressure. The bacteria contained a 2.5 kb plasmid and were sensitive to 13 drugs and insensitive to 7 drugs of the 20 drugs tested. It was suggested that alcohol dis-infectant fluid was easily contaminated by Bacillus amy-loliquefacien, and the bacteria was resistant to disinfectant fluids such as alcohol and 84 disinfectants.

  3. Multigeneration Cross-Contamination of Mail with Bacillus anthracis Spores.

    Science.gov (United States)

    Edmonds, Jason; Lindquist, H D Alan; Sabol, Jonathan; Martinez, Kenneth; Shadomy, Sean; Cymet, Tyler; Emanuel, Peter

    2016-01-01

    The release of biological agents, including those which could be used in biowarfare or bioterrorism in large urban areas, has been a concern for governments for nearly three decades. Previous incidents from Sverdlosk and the postal anthrax attack of 2001 have raised questions on the mechanism of spread of Bacillus anthracis spores as an aerosol or contaminant. Prior studies have demonstrated that Bacillus atrophaeus is easily transferred through simulated mail handing, but no reports have demonstrated this ability with Bacillus anthracis spores, which have morphological differences that may affect adhesion properties between spore and formite. In this study, equipment developed to simulate interactions across three generations of envelopes subjected to tumbling and mixing was used to evaluate the potential for cross-contamination of B. anthracis spores in simulated mail handling. In these experiments, we found that the potential for cross-contamination through letter tumbling from one generation to the next varied between generations while the presence of a fluidizer had no statistical impact on the transfer of material. Likewise, the presence or absence of a fluidizer had no statistically significant impact on cross-contamination levels or reaerosolization from letter opening. PMID:27123934

  4. Multigeneration Cross-Contamination of Mail with Bacillus anthracis Spores.

    Directory of Open Access Journals (Sweden)

    Jason Edmonds

    Full Text Available The release of biological agents, including those which could be used in biowarfare or bioterrorism in large urban areas, has been a concern for governments for nearly three decades. Previous incidents from Sverdlosk and the postal anthrax attack of 2001 have raised questions on the mechanism of spread of Bacillus anthracis spores as an aerosol or contaminant. Prior studies have demonstrated that Bacillus atrophaeus is easily transferred through simulated mail handing, but no reports have demonstrated this ability with Bacillus anthracis spores, which have morphological differences that may affect adhesion properties between spore and formite. In this study, equipment developed to simulate interactions across three generations of envelopes subjected to tumbling and mixing was used to evaluate the potential for cross-contamination of B. anthracis spores in simulated mail handling. In these experiments, we found that the potential for cross-contamination through letter tumbling from one generation to the next varied between generations while the presence of a fluidizer had no statistical impact on the transfer of material. Likewise, the presence or absence of a fluidizer had no statistically significant impact on cross-contamination levels or reaerosolization from letter opening.

  5. Biodegradation of malathion by Bacillus licheniformis strain ML-1

    Directory of Open Access Journals (Sweden)

    Khan Sara

    2016-01-01

    Full Text Available Malathion, a well-known organophosphate pesticide, has been used in agriculture over the last two decades for controlling pests of economically important crops. In the present study, a single bacterium, ML-1, was isolated by soil-enrichment technique and identified as Bacillus licheniformis on the basis of the 16S rRNA technique. The bacterium was grown in carbon-free minimal salt medium (MSM and was found to be very efficient in utilizing malathion as the sole source of carbon. Biodegradation experiments were performed in MSM without carbon source to determine the malathion degradation by the selected strain, and the residues of malathion were determined quantitatively using HPLC techniques. Bacillus licheniformis showed very promising results and efficiently consumed malathion as the sole carbon source via malathion carboxylesterase (MCE, and about 78% malathion was degraded within 5 days. The carboxylesterase activity was determined by using crude extract while using malathion as substrate, and the residues were determined by HPLC. It has been found that the MCE hydrolyzed 87% malathion within 96 h of incubation. Characterization of crude MCE revealed that the enzyme is robust in nature in terms of organic solvents, as it was found to be stable in various concentrations of ethanol and acetonitrile. Similarly, and it can work in a wide pH and temperature range. The results of this study highlighted the potential of Bacillus licheniformis strain ML-1 as a biodegrader that can be used for the bioremediation of malathion-contaminated soil.

  6. Multigeneration Cross-Contamination of Mail with Bacillus anthracis Spores

    Science.gov (United States)

    Edmonds, Jason; Lindquist, H. D. Alan; Sabol, Jonathan; Martinez, Kenneth; Shadomy, Sean; Cymet, Tyler; Emanuel, Peter

    2016-01-01

    The release of biological agents, including those which could be used in biowarfare or bioterrorism in large urban areas, has been a concern for governments for nearly three decades. Previous incidents from Sverdlosk and the postal anthrax attack of 2001 have raised questions on the mechanism of spread of Bacillus anthracis spores as an aerosol or contaminant. Prior studies have demonstrated that Bacillus atrophaeus is easily transferred through simulated mail handing, but no reports have demonstrated this ability with Bacillus anthracis spores, which have morphological differences that may affect adhesion properties between spore and formite. In this study, equipment developed to simulate interactions across three generations of envelopes subjected to tumbling and mixing was used to evaluate the potential for cross-contamination of B. anthracis spores in simulated mail handling. In these experiments, we found that the potential for cross-contamination through letter tumbling from one generation to the next varied between generations while the presence of a fluidizer had no statistical impact on the transfer of material. Likewise, the presence or absence of a fluidizer had no statistically significant impact on cross-contamination levels or reaerosolization from letter opening. PMID:27123934

  7. Effect of supplemental Bacillus culture on rumen fermentation and performance in dairy cattle

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Two parts were involved in this experiment. In experiment 1, 32 Chinese Holstein cows with relatively similar body condition, lactation number and days in milk were selected. The cows were assigned in a randomized complete block design trial to determine the effect of supplemental Bacillus cultures to diet on production performance in dairy cattle. Four treatments, i.e., Bacillus licheniformis (strain number 1.813) group, Bacillus subtilis (strain number 1.1086) group, Bacillus cereus var. mycoides (strain number 1.260) group and control group. Each treatment had eight replicates, each replicate had one cow, 50 g per head per day. Results showed that Bacillus licheniformis group increased the milk yield (P0.05). In experiment 2, 3 Chinese Holstein cows with permanent fistulas were used. 3×3 Latin squares were assigned to three diets: Bacillus lincheniformis culture, Bacillus subtilis culture and control. Bacillus licheniformis culture increased total rumen microorganism (P0.05), increased the rate of acetic acid to propionic acid (P>0.05). Bacillus licheniformis culture decreased the methane production (P>0.05).

  8. Bacillus subtilis HJ18-4 from traditional fermented soybean food inhibits Bacillus cereus growth and toxin-related genes.

    Science.gov (United States)

    Eom, Jeong Seon; Lee, Sun Young; Choi, Hye Sun

    2014-11-01

    Bacillus subtilis HJ18-4 isolated from buckwheat sokseongjang, a traditional Korean fermented soybean food, exhibits broad-spectrum antimicrobial activity against foodborne pathogens, including Bacillus cereus. In this study, we investigated the antibacterial efficacy and regulation of toxin gene expression in B. cereus by B. subtilis HJ18-4. Expression of B. cereus toxin-related genes (groEL, nheA, nheC, and entFM) was downregulated by B. subtilis HJ18-4, which also exhibited strong antibacterial activity against B. cereus. We also found that water extracts of soy product fermented with B. subtilis HJ18-4 significantly inhibited the growth of B. cereus and toxin expression. These results indicate that B. subtilis HJ18-4 could be used as an antimicrobial agent to control B. cereus in the fermented soybean food industry. Our findings also provide an opportunity to develop an efficient biological control agent against B. cereus.

  9. Genetic relationships between sympatric populations of Bacillus cereus and Bacillus thuringiensis, as revealed by rep-PCR genomic fingerprinting

    Directory of Open Access Journals (Sweden)

    Ana Paula S Peruca

    2008-08-01

    Full Text Available The bacterial strain Bacillus cereus is closely related to Bacillus thuringiensis, although any genetic relationship between the two strains is still in debate. Using rep-PCR genomic fingerprinting, we established the genetic relationships between Brazilian sympatric populations of B. cereus and B. thuringiensis simultaneously collected from two geographically separate sites. We observed the formation of both B. thuringiensis and B. cereus clusters, as well as strains of B. cereus that are more closely related to B. thuringiensis than to other B. cereus strains. In addition, lower genetic variability was observed among B. thuringiensis clusters compared to B. cereus clusters, indicating that either the two species should be categorized as separate or that B. thuringiensis may represent a clone from a B. cereus background.

  10. Bacillus thuringiensis: general characteristics and fermentation
    Bacillus thuringiensis: características gerais e fermentação

    OpenAIRE

    Raúl Jorge Hernan Castro-Gómez; Gislayne Trindade Vilas-Bôas; Elisangela Andrade Angelo

    2010-01-01

    The insect control is carried out mostly by chemical products, whose cumulative effects cause serious losses to environmental and human health, highlighting rapid selection of resistant insects. Biological control by entomopathogenic bacteria is an efficient alternative, mainly due to high specificity, absence of resistance in the target insects and low environment residual effect. Bacillus thuringiensis is a Gram-positive spore-forming bacterium that produces a parasporal crystal protein tox...

  11. Bacillus anthracis Virulent Plasmid pX02 Genes Found in Large Plasmids of Two Other Bacillus Species

    OpenAIRE

    Luna, Vicki A.; King, Debra S.; Peak, K. Kealy; Reeves, Frank; Heberlein-Larson, Lea; Veguilla, William; Heller, L.; Duncan, Kathleen E; Cannons, Andrew C.; Amuso, Philip; Cattani, Jacqueline

    2006-01-01

    In order to cause the disease anthrax, Bacillus anthracis requires two plasmids, pX01 and pX02, which carry toxin and capsule genes, respectively, that are used as genetic targets in the laboratory detection of the bacterium. Clinical, forensic, and environmental samples that test positive by PCR protocols established by the Centers for Disease Control and Prevention for B. anthracis are considered to be potentially B. anthracis until confirmed by culture and a secondary battery of tests. We ...

  12. Effects of Electrolyzed Oxidizing Water on Inactivation of Bacillus subtilis and Bacillus cereus Spores in Suspension and on Carriers.

    Science.gov (United States)

    Zhang, Chunling; Li, Baoming; Jadeja, Ravirajsinh; Hung, Yen-Con

    2016-01-01

    Spores of some Bacillus species are responsible for food spoilage and foodborne disease. These spores are highly resistant to various interventions and cooking processes. In this study, the sporicidal efficacy of acidic electrolyzed oxidizing (EO) water (AEW) and slightly acidic EO water (SAEW) with available chlorine concentration (ACC) of 40, 60, 80, 100, and 120 mg/L and treatment time for 1, 2, 3, 4, 5, and 6 min were tested on Bacillus subtilis and Bacillus cereus spores in suspension and on carrier with or without organics. The reduction of spore significantly increased with increasing ACC and treatment time (P waters containing 120 mg/L ACC, while only SAEW at 120 mg/L and 2 min treatment achieved >6 log reductions of B. subtilis spore. Both types of EO water with ACC of 60 mg/L and 6 min treatment achieved a reduction of B. subtilis and B. cereus spores to nondetectable level. EO water with ACC of 80 mg/L and treatment time of 3 min on carrier test without organics addition resulted in reductions of B. subtilis spore to nondetectable level. But, addition of 0.3% organics on carrier decreased the inactivation effect of EO water. This study indicated that EO water was highly effective in inactivation of B. subtilis and B. cereus spores in suspension or on carrier, and therefore, rendered it as a promising disinfectant to be applied in food industry.

  13. Regulation of the tryptophan biosynthetic genes in Bacillus halodurans: common elements but different strategies than those used by Bacillus subtilis.

    Science.gov (United States)

    Szigeti, Reka; Milescu, Mirela; Gollnick, Paul

    2004-02-01

    In Bacillus subtilis, an RNA binding protein called TRAP regulates both transcription and translation of the tryptophan biosynthetic genes. Bacillus halodurans is an alkaliphilic Bacillus species that grows at high pHs. Previous studies of this bacterium have focused on mechanisms of adaptation for growth in alkaline environments. We have characterized the regulation of the tryptophan biosynthetic genes in B. halodurans and compared it to that in B. subtilis. B. halodurans encodes a TRAP protein with 71% sequence identity to the B. subtilis protein. Expression of anthranilate synthetase, the first enzyme in the pathway to tryptophan, is regulated significantly less in B. halodurans than in B. subtilis. Examination of the control of the B. halodurans trpEDCFBA operon both in vivo and in vitro shows that only transcription is regulated, whereas in B. subtilis both transcription of the operon and translation of trpE are controlled. The attenuation mechanism that controls transcription in B. halodurans is similar to that in B. subtilis, but there are some differences in the predicted RNA secondary structures in the B. halodurans trp leader region, including the presence of a potential anti-antiterminator structure. Translation of trpG, which is within the folate operon in both bacilli, is regulated similarly in the two species. PMID:14729709

  14. Genome Sequence of the Endophytic Bacterium Bacillus thuringiensis Strain KB1, a Potential Biocontrol Agent against Phytopathogens

    OpenAIRE

    Jeong, Haeyoung; Jo, Sung Hee; Hong, Chi Eun; Park, Jeong Mee

    2016-01-01

    Bacillus thuringiensis is the most widely known microbial pesticide used in agricultural applications. Herein, we report a draft genome sequence of the endophytic bacterium Bacillus thuringiensis strain KB1, which exhibits antagonism against phytopathogens.

  15. Genome Sequence of the Endophytic Bacterium Bacillus thuringiensis Strain KB1, a Potential Biocontrol Agent against Phytopathogens.

    Science.gov (United States)

    Jeong, Haeyoung; Jo, Sung Hee; Hong, Chi Eun; Park, Jeong Mee

    2016-04-21

    ITALIC! Bacillus thuringiensisis the most widely known microbial pesticide used in agricultural applications. Herein, we report a draft genome sequence of the endophytic bacterium ITALIC! Bacillus thuringiensisstrain KB1, which exhibits antagonism against phytopathogens.

  16. Two purine nucleoside phosphorylases in Bacillus subtilis. Purification and some properties of the adenosine-specific phosphorylase

    DEFF Research Database (Denmark)

    Jensen, Kaj Frank

    1978-01-01

    Two purine nucleoside phosphorylases (purine-nucleoside:orthophosphate ribosyltransferase, EC 2.4.2.1) were purified from vegetative Bacillus subtilis cells. One enzyme, inosine-guanosine phosphorylase, showed great similarity to the homologous enzyme of Bacillus cereus. It appeared...

  17. Resistance to antimicrobials and acid and bile tolerance of Bacillus spp isolated from Bikalga, fermented seeds of Hibiscus sabdariffa

    OpenAIRE

    Compaore, Clarisse S.; Jensen, Lars Bogø; Diawara, Brehima; Ouedraogo, Georges A.; Jakobsen, Mogens; Ouoba, Labia I. I.

    2013-01-01

    In the aim of selecting starter cultures, thirteen species of Bacillus spp. including six Bacillus subtilis ssp. subtilis, four Bacillus licheniformis and three Bacillus amyloliquefaciens ssp. plantarum isolated from traditional Bikalga were investigated. The study included, for all isolates, genes, determination of minimal inhibitory concentration (MIC) for 24 antimicrobials and detection of resistance by PCR using specific primers. The isolates were also examined for their resistance to pH ...

  18. Isolation and Identification of Bacillus Species From Soil and Evaluation of Their Antibacterial Properties

    Directory of Open Access Journals (Sweden)

    Amin

    2015-02-01

    Full Text Available Background Bacillus species are the predominant soil bacteria because of their resistant-endospore formation and production of essential antibiotics such as bacitracin. Objectives The aim of this study was to isolate Bacillus spp. from riverside soil and investigate their antimicrobial characteristics against some pathogenic bacteria. Materials and Methods Fifty soil samples were collected from different sites of Bahmanshir riverside in Abadan city, Iran, and analyzed for the presence of Bacillus species. The media used in this research were nutrient broth and agar. Bacillus species were identified by their phenotypic and biochemical characteristics. The antimicrobial effects of Bacillus extract against the target bacteria including Escherichia coli, Staphylococcus aureus, Salmonella typhi, Shigella dysenteriae and Corynebacterium diphtheriae were examined. Results The identified Bacillus species included B. cereus (86.6%, B. subtilis (6.6%, B. thuringiensis (3.3%, and B. pumilus (3.3%. Evaluation of the antimicrobial activity of the extracted compounds was carried out against five different bacteria. Antibiotic production tests indicated that two Bacillus strains belong to B. cereus, which showed antimicrobial properties. The minimum inhibitory concentrations (MICs of these compounds ranged between 8.34-33.34 mg/mL for the target bacteria. Conclusions This study indicated that some Bacillus species have the potential to produce antimicrobial compounds which can be used to control microbial infections.

  19. The structure-function relationship of the lipases from Pseudomonas aeruginosa and Bacillus subtilis

    NARCIS (Netherlands)

    Misset, Onno; Gerritse, Gijs; Jaeger, Karl-Erich; Winkler, Ulrich; Colson, Charles; Schanck, Karin; Lesuisse, Emmanuel; Dartois, Véronique; Blaauw, Mieke; Ransac, Stéphane; Dijkstra, Bauke W.

    1994-01-01

    Within the BRIDGE T-project on lipases we investigate the structure-function relationships of the lipases from Bacillus subtilis and Pseudomonas aeruginosa. Construction of an overproducing Bacillus strain allowed the purification of > 100 mg lipase from 30 I culture supernatant. After testing a lar

  20. Lead biotransformation potential of allochthonous Bacillus sp. SKK11 with sesame oil cake in mine soil

    Science.gov (United States)

    This study was aimed at assessing the potential of allochthonous Bacillus sp. SKK11 and sesame oil cake extract for transformation of Pb in mine soil. The bacteria were isolated from a brackish environment and identified as Bacillus sp. based on partial 16S rDNA sequences. The isolate SKK11 exhibite...

  1. PCR detection of cytK gene in Bacillus cereus group strains isolated from food samples.

    Science.gov (United States)

    Oltuszak-Walczak, Elzbieta; Walczak, Piotr

    2013-11-01

    A method for detection of the cytotoxin K cytK structural gene and its active promoter preceded by the PlcR-binding box, controlling the expression level of this enterotoxin, was developed. The method was applied for the purpose of the analysis of 47 bacterial strains belonging to the Bacillus cereus group isolated from different food products. It was found that the majority of the analyzed strains carried the fully functional cytK gene with its PlcR regulated promoter. The cytK gene was not detected in four emetic strains of Bacillus cereus carrying the cesB gene and potentially producing an emetic toxin - cereulide. The cytotoxin K gene was detected in 4 isolates classified as Bacillus mycoides and one reference strain B. mycoides PCM 2024. The promoter region and the N-terminal part of the cytK gene from two strains of B. mycoides (5D and 19E) showed similarities to the corresponding sequences of Bacillus cereus W23 and Bacillus thuringiensis HD-789, respectively. It was shown for the first time that the cytK gene promoter region from strains 5D and 19E of Bacillus mycoides had a similar arrangement to the corresponding sequence of Bacillus cereus ATCC 14579. The presence of the cytK gene in Bacillus mycoides shows that this species, widely recognized as nonpathogenic, may pose potential biohazard to human beings.

  2. Diversity of Secondary Metabolites from Marine Bacillus Species: Chemistry and Biological Activity

    OpenAIRE

    Hee Jae Shin; Muhammad Abdul Mojid Mondol; Mohammad Tofazzal Islam

    2013-01-01

    Marine Bacillus species produce versatile secondary metabolites including lipopeptides, polypeptides, macrolactones, fatty acids, polyketides, and isocoumarins. These structurally diverse compounds exhibit a wide range of biological activities, such as antimicrobial, anticancer, and antialgal activities. Some marine Bacillus strains can detoxify heavy metals through reduction processes and have the ability to produce carotenoids. The present article reviews the chemistry and biological activi...

  3. Transformation of Bacillus thuringiensis subsp. galleria protoplasts by plasmid pBC16.

    OpenAIRE

    Alikhanian, S. I.; Ryabchenko, N F; Bukanov, N O; Sakanyan, V A

    1981-01-01

    Protoplasts of the entomopathogenic bacterium Bacillus thuringiensis subsp. galleria were transformed by plasmid pBC16. The frequency of transformation was much lower than that of Bacillus subtilis. All isolated B. thuringiensis transformants were characterized by increased sensitivity to lysozyme as compared with the original strain.

  4. Macrophage-Enhanced Germination of Bacillus anthracis Endospores Requires gerS

    OpenAIRE

    Ireland, John A. W.; Hanna, Philip C.

    2002-01-01

    Germination of Bacillus anthracis Sterne and plasmidless Δ-Sterne endospores was dramatically enhanced in RAW264.7 macrophage-like cells, while germination of nonpathogenic Bacillus endospores was not. Elimination of gerS, a germinant receptor locus, caused a complete loss of cell-enhanced germination, implicating gerS in the breaking of endospore dormancy in vivo.

  5. Evaluation of bottlenecks in the late stages of protein secretion in Bacillus subtilis

    NARCIS (Netherlands)

    Bolhuis, A; Tjalsma, H; Smith, H.E; Meima, R.; Venema, G; Bron, S; van Dijl, J.M

    1999-01-01

    Despite a high capacity for secretion of homologous proteins, the secretion of heterologous proteins by Bacillus subtilis is frequently inefficient. In the present studies, we have investigated and compared bottlenecks in the secretion of four heterologous proteins: Bacillus lichenifomis alpha-amyla

  6. Characterization of Emetic Bacillus weihenstephanensis, a New Cereulide-Producing Bacterium

    DEFF Research Database (Denmark)

    Thorsen, Line; Munk Hansen, Bjarne; Nielsen, Kristian Fog;

    2006-01-01

    Cereulide production has until now been restricted to the species Bacillus cereus. Here we report on two psychrotolerant Bacillus weihenstephanensis strains, MC67 and MC118, that produce cereulide. The strains are atypical with regard to pheno- and genotypic characteristics normally used...

  7. Engineering of thermotolerant Bacillus coagulans for production of D(-)-lactic acid

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Qingzhao; Shanmugam, Keelnatham T; Ingram, Lonnie O

    2014-12-02

    Genetically modified microorganisms having the ability to produce D(-)-lactic acid at temperatures between 30.degree. C. and 55.degree. C. are provided. In various embodiments, the microorganisms may have the chromosomal lactate dehydrogenase (ldh) gene and/or the chromosomal acetolactate synthase (alsS) gene inactivated. Exemplary microorganisms for use in the disclosed methods are Bacillus spp., such as Bacillus coagulans.

  8. SR450 and Superhawk XP applications of Bacillus thuringiensis israelensis de Barjac against Culex quinquefasciatus Say

    Science.gov (United States)

    Sprayer comparisons and larval morality assays were conducted following SR450 backpack mist blower and Superhawk XP thermal fogger applications of Vectobac® WDG Bacillus thuringiensis israelensis (Bti) de Barjac against Culex quinquefasciatus Say. Bacillus thuringiensis israelensis was applied at m...

  9. [Characterization of crystal-forming bacteria Bacillus thuringiensis subsp. tohokuensis toxic to mosquitos].

    Science.gov (United States)

    Khodyrev, V P; Kalmykova, G V; Burtseva, L I; Glupov, V V

    2006-01-01

    Distribution study of Bacillus thuringiensis strains in Western Siberian soils allowed us to isolate crystal-forming bacteria assigned to a new pathovar of Bacillus thuringiensis ssp. tohokuensis with a toxic effect on mosquito larvae. A description of this bacterial pathovar is presented.

  10. Draft Whole-Genome Sequence of the Type Strain Bacillus horikoshii DSM 8719

    Science.gov (United States)

    Hernández-González, Ismael L.

    2016-01-01

    Members of the Bacillus genus have been extensively studied because of their ability to produce enzymes with high biotechnological value. Here, we report the draft of the whole-genome sequence of the type strain Bacillus horikoshii DSM 8719, an alkali-tolerant strain. PMID:27417833

  11. Role of enzymes of homologous recombination in illegitimate plasmid recombination in Bacillus subtilis

    NARCIS (Netherlands)

    Meima, R; Haijema, BJ; Haan, GJ; Venema, G; Bron, S

    1997-01-01

    The structural stability of plasmid pGP1, which encodes a fusion between the penicillinase gene (penP) of Bacillus licheniformis and the Escherichia coli lacZ gene, was investigated in Bacillus subtilis strains expressing mutated subunits of the ATP-dependent nuclease, AddAB, and strains lacking the

  12. The expression of a plasmid-specified exported protein causes structural plasmid instability in Bacillus subtilis

    NARCIS (Netherlands)

    Cordes, C.; Meima, R; Twiest, B; Kazemier, B; Venema, G; vanDijl, JM; Bron, S

    1996-01-01

    The rolling-circle plasmid pGP1 was used to study the effects of the expression of a plasmid-specified exported protein on structural plasmid stability in Bacillus subtilis. pGP1 contains a fusion between the Bacillus licheniformis penP gene, encoding a C-terminally truncated penicillinase, and the

  13. In Vitro Assessment of Marine Bacillus for Use as Livestock Probiotics

    Directory of Open Access Journals (Sweden)

    Maria Luz Prieto

    2014-04-01

    Full Text Available Six antimicrobial-producing seaweed-derived Bacillus strains were evaluated in vitro as animal probiotics, in comparison to two Bacillus from an EU-authorized animal probiotic product. Antimicrobial activity was demonstrated on solid media against porcine Salmonella and E. coli. The marine isolates were most active against the latter, had better activity than the commercial probiotics and Bacillus pumilus WIT 588 also reduced E. coli counts in broth. All of the marine Bacillus tolerated physiological concentrations of bile, with some as tolerant as one of the probiotics. Spore counts for all isolates remained almost constant during incubation in simulated gastric and ileum juices. All of the marine Bacillus grew anaerobically and the spores of all except one isolate germinated under anaerobic conditions. All were sensitive to a panel of antibiotics and none harbored Bacillus enterotoxin genes but all, except B. pumilus WIT 588, showed some degree of β-hemolysis. However, trypan blue dye exclusion and xCELLigence assays demonstrated a lack of toxicity in comparison to two pathogens; in fact, the commercial probiotics appeared more cytotoxic than the majority of the marine Bacillus. Overall, some of the marine-derived Bacillus, in particular B. pumilus WIT 588, demonstrate potential for use as livestock probiotics.

  14. Genome Sequence of the Plant Growth-Promoting Rhizobacterium Bacillus sp. Strain 916

    OpenAIRE

    Wang, Xiaoyu; Luo, Chuping; Chen, Zhiyi

    2012-01-01

    Bacillus sp. strain 916, isolated from the soil, showed strong activity against Rhizoctonia solani. Here, we present the high-quality draft genome sequence of Bacillus sp. strain 916. Its 3.9-Mb genome reveals a number of genes whose products are possibly involved in promotion of plant growth or antibiosis.

  15. Bacillus cereus in Brazilian Ultra High Temperature milk Bacillus cereus em leite UHT brasileiro

    Directory of Open Access Journals (Sweden)

    Cristiana de Paula Pacheco-Sanchez

    2007-01-01

    Full Text Available Brazilian Ultra High Temperature (UHT milk consumption has increased during the last decade from 187 to 4,200 million liters. In the continuous UHT process, milk is submitted for 2-4 s to 130-150ºC, in a continuous flow system with immediate refrigeration and aseptical packing in hermetic packages. This research had the purpose to verify the incidence of B. cereus species from the B. cereus group, in UHT milk. In 1998 high indexes of these organisms were reported, reaching 34.14% of the analyzed samples. Beyond this fact, there was the need to establish methods and processes adjusted for correct identification of B. cereus. Thus, commercial sterility tests of 6,500 UHT milk packages were investigated in two assays, after ten days incubation at 37ºC and 7ºC to germinate all possible spores and/or to recuperate injured vegetative cells followed by pH measurement. Samples (1,300 packages each from five Brazilian UHT plants of whole UHT milk processed by direct steam injection, packaged in carton were investigated for the presence of Bacillus cereus through phenotypic and genetic (PCR tests. Values of pH were different for the samples, ranging between 6.57 and 6.73. After storage of the samples, only four packages with pH measurement below the lower limit of 6.5 were found and analyzed for the presence of B. cereus. This organism was not detected in any of the samples indicating that the five Brazilian UHT milk processors control pathogenic microorganisms and it can be said that the consumption of UHT milk does not present safety problems to consumers. Fourier Transform Infrared Spectroscopy (FTIR and PCR tests were efficient and must be adopted to confirm the biochemical series for B. cereus.O consumo de leite ultra-alta temperatura (UHT brasileiro aumentou, durante a última década, de 187 milhões de litros para 4,200 milhões de litros. No processo contínuo de leite UHT o leite é submetido por 2-4 seg a 130-150ºC, em sistemas de

  16. Bacillus vanillea sp. nov., Isolated from the Cured Vanilla Bean.

    Science.gov (United States)

    Chen, Yong-gan; Gu, Feng-lin; Li, Ji-hua; Xu, Fei; He, Shu-zhen; Fang, Yi-ming

    2015-02-01

    A Gram-positive bacterium, designated strain XY18(T), was isolated from a cured vanilla bean in Hainan province, China. Cells were rod-shaped, endospore producing, and peritrichous flagella. Strain XY18(T) grew at salinities of 0-8 % (w/v) NaCl (optimally 1-4 %), pH 4.0-8.0 (optimally 5.0-7.0 %) and temperature range 20-45 °C (optimally 28-35 °C). The predominant menaquinone was MK-7. The major cellular fatty acids were anteiso-C15:0, iso-C15:0, anteiso-C17:0, and iso-C17:0. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain XY18(T) was a member of the genus Bacillus, and closely related to B. amyloliquefaciens NBRC 15535(T) and B. siamensis PD-A10(T), with 99.1 and 99.2 % sequence similarity, respectively. However, the DNA-DNA hybridization value between strain XY18(T) and B. amyloliquefaciens NBRC 15535(T) was 35.7 %. The genomic DNA G+C content of strain XY18(T) was 46.4 mol%, significantly differed from B. siamensis PD-A10(T) (41.4 %), which was higher than the range of 4 % indicative of species. On the basis of polyphasic taxonomic study, including phenotypic features, chemotaxonomy, and phylogenetic analyses, strain XY18(T) represents a novel species within the genus Bacillus, for which the name Bacillus vanillea sp. nov. is proposed. The type strain is XY18(T) (=CGMCC 8629 = NCCB 100507).

  17. Application of Bacillus sp. as a biopreservative for food preservation

    Directory of Open Access Journals (Sweden)

    S. Nath,

    2015-04-01

    Full Text Available Food preservation is enhancing shelf-life and food quality to eliminate food-related illness and product spoilage, especially by the use of food additives.The growing consumer demand for effective preservation of food without altering its nutritional quality and free of potential health risks andto find an attractive and alternative approach to chemical preservatives, have stimulated research in the field of biopreservation by the use of natural or controlled microbiota and/or their antimicrobial compounds including very recent innovation: Bacillus sp., the ubiquitous, Gram positive bacteria, producing inhibitory substances like cyclic peptides and bacteriocins, with a broad antimicrobial spectrum and a history of safe use in food. Bacillus spores are also being used extensively as probiotic food supplements where they are used in human as dietary supplements and in feed for livestock and aquaculture as growth promoters.A novel concept multi-target food preservation has emerged in relation to hurdle technology stating the microbial safety, stability, sensorial and nutritional qualities of foods are based on the application of combined preservative factors (called hurdles including Bacillus sp. that microorganisms present in the food are unable to overcome, thus leading to inhibition of microbial growth by disturbing their homeostasis and metabolic exhaustion and avoiding tress reaction by bacteria. Future exploration of the natural preservatives and/or their metabolites, in combination with advanced technologies could result in replacement of chemical preservatives, or could allow less severe processing (e.g. heat treatments, while still maintaining adequate microbiological safety and quality in foods.

  18. Bacillus vanillea sp. nov., Isolated from the Cured Vanilla Bean.

    Science.gov (United States)

    Chen, Yong-gan; Gu, Feng-lin; Li, Ji-hua; Xu, Fei; He, Shu-zhen; Fang, Yi-ming

    2015-02-01

    A Gram-positive bacterium, designated strain XY18(T), was isolated from a cured vanilla bean in Hainan province, China. Cells were rod-shaped, endospore producing, and peritrichous flagella. Strain XY18(T) grew at salinities of 0-8 % (w/v) NaCl (optimally 1-4 %), pH 4.0-8.0 (optimally 5.0-7.0 %) and temperature range 20-45 °C (optimally 28-35 °C). The predominant menaquinone was MK-7. The major cellular fatty acids were anteiso-C15:0, iso-C15:0, anteiso-C17:0, and iso-C17:0. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain XY18(T) was a member of the genus Bacillus, and closely related to B. amyloliquefaciens NBRC 15535(T) and B. siamensis PD-A10(T), with 99.1 and 99.2 % sequence similarity, respectively. However, the DNA-DNA hybridization value between strain XY18(T) and B. amyloliquefaciens NBRC 15535(T) was 35.7 %. The genomic DNA G+C content of strain XY18(T) was 46.4 mol%, significantly differed from B. siamensis PD-A10(T) (41.4 %), which was higher than the range of 4 % indicative of species. On the basis of polyphasic taxonomic study, including phenotypic features, chemotaxonomy, and phylogenetic analyses, strain XY18(T) represents a novel species within the genus Bacillus, for which the name Bacillus vanillea sp. nov. is proposed. The type strain is XY18(T) (=CGMCC 8629 = NCCB 100507). PMID:25292250

  19. Food – bacteria interplay: Pathometabolism of emetic Bacillus cereus

    Directory of Open Access Journals (Sweden)

    Monika eEhling-Schulz

    2015-07-01

    Full Text Available Bacillus cereus is a gram-positive endospore forming bacterium known for its wide spectrum of phenotypic traits, enabling it to occupy diverse ecological niches. Although the population structure of B. cereus is highly dynamic and rather panmictic, production of the emetic B. cereus toxin cereulide is restricted to strains with specific genotypic traits, associated with distinct environmental habitats. Cereulide is an ionophoric dodecadepsipeptide that is produced non-ribosomally by an enzyme complex with an unusual modular structure, named cereulide synthetase (Ces NRPS. The ces gene locus is encoded on a mega virulence plasmid related to the Bacillus anthracis toxin plasmid pXO1. Cereulide, a highly thermo- and pH- resistant molecule, is preformed in food, evokes vomiting a few hours after ingestion and was shown to be the direct cause of gastroenteritis symptoms; occasionally it is implicated in severe clinical manifestations including acute liver failures. Control of toxin gene expression in emetic Bacillus cereus involves central transcriptional regulators, such as CodY and AbrB, thereby inextricably linking toxin gene expression to life cycle phases and specific conditions, such as the nutrient supply encountered in food matrices. While in recent years considerable progress has been made in the molecular and biochemical characterization of cereulide toxin synthesis, far less is known about the embedment of toxin synthesis in the life cycle of B. cereus. Information about signals acting on toxin production in the food environment is literally lacking. We summarize the data available on the complex regulatory network controlling cereulide toxin synthesis, discuss the role of intrinsic and extrinsic factors acting on toxin biosynthesis in emetic B. cereus and stress how unraveling these processes can lead to the development of novel effective strategies to prevent toxin synthesis in the food production and processing chain.

  20. Application of Bacillus sphaericus in the control of Culex fatigans

    International Nuclear Information System (INIS)

    A terminal spore bearing bacteria strain (ISPC-5) was isolated from the diseased larvae of Culex fatigans and identified as Bacillus sphaericus (WHO 2173). It was of the phage type IV and serotype H-26a and 26b. The LC50 for C. fatigans 2nd instar larvae was 1.7x104 colony forming units (CFU)/mL. Comparative toxicity studies made on ISPC-5, as well as on 1593 and Bacillus thuringiensis var. israelensis (H-14), revealed that this organism was pathogenic to laboratory reared non-resistant and chemical insecticide resistant species of the Culex genus, C. fatigans in particular. However, this organism was found to be non-pathogenic to Anopheline and Aedine larval instars. The spore stage of this bacillus is affected after exposure to sunlight for 6 hours and to UV germicidal radiation for 120 minutes at a dose of 108x103J/m2, and tolerates heat treatment at 60 deg. C for 30 minutes only. In all cases the viability and toxicity are drastically affected. It is non-toxic to Gambusia affinis. Small and large scale laboratory trials with C. fatigans larval instars produced good results. The field trials conducted in the Bombay suburbs in septic tanks with a concentration of 105 CFU/mL proved encouraging. Spores of this organism have a good shelf-life in a cold room (-10 deg. C) or at room temperature as lyophilized material. This indigenously isolated B. sphaericus (WHO 2173) can successfully be used per se in controlling C. fatigans or in integrated vector control programmes. (author). 18 refs, 5 tabs

  1. Assessment of a new selective chromogenic Bacillus cereus group plating medium and use of enterobacterial autoinducer of growth for cultural identification of Bacillus species.

    Science.gov (United States)

    Reissbrodt, R; Rassbach, A; Burghardt, B; Rienäcker, I; Mietke, H; Schleif, J; Tschäpe, H; Lyte, M; Williams, P H

    2004-08-01

    A new chromogenic Bacillus cereus group plating medium permits differentiation of pathogenic Bacillus species by colony morphology and color. Probiotic B. cereus mutants were distinguished from wild-type strains by their susceptibilities to penicillin G or cefazolin. The enterobacterial autoinducer increased the sensitivity and the speed of enrichment of B. cereus and B. anthracis spores in serum-supplemented minimal salts medium (based on the standard American Petroleum Institute medium) and buffered peptone water. PMID:15297532

  2. Enterotoxins and emetic toxins production by Bacillus cereus and other species of Bacillus isolated from Soumbala and Bikalga, African alkaline fermented food condiments.

    Science.gov (United States)

    Ouoba, Labia Irene I; Thorsen, Line; Varnam, Alan H

    2008-06-10

    The ability of various species of Bacillus from fermented seeds of Parkia biglobosa known as African locust bean (Soumbala) and fermented seeds of Hibiscus sabdariffa (Bikalga) was investigated. The study included screening of the isolates by haemolysis on blood agar, detection of toxins in broth and during the fermentation of African locust bean using the Bacillus cereus Enterotoxin Reverse Passive Latex Agglutination test kit (BCET-RPLA) and the Bacillus Diarrhoeal Enterotoxin Visual Immunoassay (BDEVIA). Detection of genes encoding cytotoxin K (CytK), haemolysin BL (Hbl A, Hbl C, Hbl D), non-hemolytic enterotoxin (NheA, NheB, NheC) and EM1 specific of emetic toxin producers was also investigated using PCR with single pair and multiplex primers. Of 41 isolates, 29 Bacillus belonging to the species of B. cereus, Bacillus subtilis, Bacillus licheniformis and Bacillus pumilus showed haemolysis on blood agar. Using RPLA, enterotoxin production was detected for three isolates of B. cereus in broth and all B. cereus (9) in fermented seeds. Using BDEVIA, enterotoxin production was detected in broth as well as in fermented seeds for all B. cereus isolates. None of the isolates belonging to the other Bacillus species was able to produce enterotoxins either by RPLA or BDEVIA. Nhe genes were detected in all B. cereus while Hbl and CytK genes were detected respectively in five and six B. cereus strains. A weak presence of Hbl (A, D) and CytK genes was detected in two isolates of B. subtilis and one of B. licheniformis but results were inconsistent, especially for Hbl genes. The emetic specific gene fragment EM1 was not detected in any of the isolates studied.

  3. Effect of Bacillus thuringiensis on microbial functional groups in sorghum rhizosphere Efeito do Bacillus thuringiensis sobre grupos funcionais de microrganismos na rizosfera de sorgo

    OpenAIRE

    Carlos Brasil; Leopoldo Sussumu Matsumoto; Marco Antonio Nogueira; Flavia Regina Spago; Luís Gustavo Rampazo; Marcio Ferreira Cruz; Galdino Andrade

    2006-01-01

    The objective of this work was to assess the effect of two strains of Bacillus thuringiensis var. kurstaki on sorghum rhizosphere microorganisms. The strains were HD1, that produces the bioinsecticidal protein, and 407, that is a mutant non-producer. The strains do not influence microbial population, but reduce plant growth and improve mycorrhizal colonization and free living fixing N2 community.O objetivo deste trabalho foi avaliar o efeito de duas cepas de Bacillus thuringiensis var. kursta...

  4. Cosegregation of cell wall and DNA in Bacillus subtilis.

    OpenAIRE

    Schlaeppi, J M; Karamata, D

    1982-01-01

    Cosegregation of cell wall and DNA of a lysis-negative mutant of Bacillus subtilis was examined by continuously labeling (i) cell wall, (ii) DNA, and (iii) both cell wall and DNA. After four to five generations of chase in liquid media it was found by light microscope autoradiography that the numbers of wall segregation units per cell are 29 and 9 in rich and minimal medium, respectively. Under the same conditions the numbers of segregation units of DNA were almost 50% lower: 15 and 5, respec...

  5. Secretion Genes as Determinants of Bacillus anthracis Chain Length

    OpenAIRE

    Nguyen-Mau, Sao-Mai; Oh, So-Young; Kern, Valerie J.; Missiakas, Dominique M.; Schneewind, Olaf

    2012-01-01

    Bacillus anthracis grows in chains of rod-shaped cells, a trait that contributes to its escape from phagocytic clearance in host tissues. Using a genetic approach to search for determinants of B. anthracis chain length, we identified mutants with insertional lesions in secA2. All isolated secA2 mutants exhibited an exaggerated chain length, whereas the dimensions of individual cells were not changed. Complementation studies revealed that slaP (S-layer assembly protein), a gene immediately dow...

  6. Genetic analysis of Bacillus stearothermophilus by protoplast fusion.

    OpenAIRE

    Chen, Z F; Wojcik, S F; Welker, N E

    1986-01-01

    Efficient and reliable protoplasting, regeneration, and fusion techniques were established for the prototrophic strain Bacillus stearothermophilus NUB36. Auxotrophic mutants were isolated, and protoplast fusion was used to construct isogenic mutant strains and for chromosomal mapping. Markers were mapped using two-, three-, and four-factor crosses. The order of the markers was hom-1-thr-1-his-1-(gly-1 or gly-2)-pur-1-pur-2. These markers may be analogous to hom, thrA, hisA, glyC, and purA mar...

  7. Production, Secretion and Biological Activity of Bacillus cereus Enterotoxins

    Directory of Open Access Journals (Sweden)

    Sonia Senesi

    2010-06-01

    Full Text Available Bacillus cereus behaves as an opportunistic pathogen frequently causing gastrointestinal diseases, and it is increasingly recognized to be responsible for severe local or systemic infections. Pathogenicity of B. cereus mainly relies on the secretion of a wide array of toxins and enzymes and also on the ability to undergo swarming differentiation in response to surface-sensing. In this report, the pathogenicity exerted by B. cereus toxins is described with particular attention to the regulatory mechanisms of production and secretion of HBL, Nhe and CytK enterotoxins.

  8. Sternal Osteomyelitis after Bacillus Calmette-Guérin Vaccination

    Science.gov (United States)

    Selvestravičius, Rolandas; Sučilienė, Elena; Saniukas, Kęstutis; Bobelytė, Odeta; Usonis, Vytautas

    2016-01-01

    Presented here is the case of a nine-month-old boy with the osteomyelitis of the upper area sternum caused by bacillus Calmette-Guerin (BCG), the Danish 1331 strain vaccine against tuberculosis. Upon examination, a swelling of approximately 2×3 cm diameter was observed in the upper sternal area. The mass was hard, fixed and sensitive to palpation with no local skin hyperaemia. Chest X-rays revealed a round mass anterior to the sternum, suggesting a diagnosis of osteomyelitis. A consequent sternal biopsy was performed and Mycobacterium bovis BCG was identified by a positive growth culture. PMID:27777704

  9. Bacillus thuringiensis resistance in Plutella - too many trees?

    Science.gov (United States)

    Crickmore, Neil

    2016-06-01

    Plutella xylostella was the first insect for which resistance to Bacillus thuringiensis was reported in the field, yet despite many studies on the nature of this resistance phenotype its genetic and molecular basis remains elusive. Many different factors have been proposed as contributing to resistance, although in many cases it has not been possible to establish a causal link. Indeed, there are so many studies published that it has become very difficult to 'see the wood for the trees'. This article will attempt to clarify our current understanding of Bt resistance in P. xylostella and consider the criteria that are used when validating a particular model. PMID:27436736

  10. Isolation of Bacillus Cereus from wounds and burns

    Directory of Open Access Journals (Sweden)

    Behzadiannejhad Gh

    1997-09-01

    Full Text Available The culture results of 203 cases with different wounds were studies; 150 of the latter were burn cases (mainly second and third degree burns, and 53 were of other types (surgical, traumatic, ect. Four subtypes of Bacillus cereus were isolated upon culture, and the different toxins produced in DHT broth with 0.1% glucose were assessed. The lethal toxin was injected intravenously to Syrian rats, none of whom died. VPR factor was assessed in the 4 subtypes. Three subtypes produced VPR in significant amounts.

  11. Ultra-violet-resistant mutants of Bacillus thuringiensis

    International Nuclear Information System (INIS)

    One of the main disadvantages of using Bacillus thuringiensis as an insecticide is that the spore and crystal preparations applied to foliage are readily washed away by rain and are inactivated by sunlight. Spores from some strains of B. thuringiensis have been shown to be highly sensitive to u.v. light. This study has demonstrated how mutants with increased resistance to u.v., isolated by successive rounds of u.v. irradiation, and additionally with increased specific pathogenicity can be isolated. These techniques should be applied to strains that are frequently used in the industrial production of B.thuringiensis toxin. (author)

  12. Unhairing animal hides using probiotic Bacteria bacillus subtilis

    OpenAIRE

    Данилкович, Анатолій Григорович; Гвоздяк, Петро Ілліч; Романюк, Оксана Олександрівна; Ковтуненко, Ольга Василівна

    2013-01-01

    The most efficient technology of processing natural raw materials into skin and fur is the use of enzyme products for soaking and liming processes. Therefore, the use of bacterial products, which produce enzymes of various functional effects, is considered to be very promising for the above mentioned processes.Soaking and liming of flint-dried rabbit hides were carried out using probiotic bacreria Bacillus subtilis on 4 samples in a laboratory centrifuge at soaking temperature 36-38°С and wor...

  13. EXPERIMENTAL-INFECTION IN MICE WITH BACILLUS-LICHENIFORMIS

    DEFF Research Database (Denmark)

    Agerholm, J.S.; Jensen, H.E.; Jensen, N.E.

    1995-01-01

    The pathogenicity of Bacillus licheniformis was assessed in normal and immunodepressed BALB/c mice. The animals were challenged intravenously with 4 x 10(7) colony forming units of B, licheniformis (ATCC 14580) and both normal and immunodepressed mice were susceptible. However, the infection was...... more severe in the immunosuppressed animals. In normal mice, lesions were restricted to the liver and kidneys, while lesions also occurred in other organs of immunodepressed mice. By crossed immunoelectrophoresis it was shown that antigens of B. licheniformis are potent immunogens, and the bacteria...

  14. Detection of Bacillus anthracis DNA by LightCycler PCR

    OpenAIRE

    Bell, Constance A.; Uhl, James R.; Hadfield, Ted L.; David, John C.; Meyer, Richard F.; Smith, Thomas F.; Cockerill III, Franklin R.

    2002-01-01

    Anthrax is a zoonotic disease that is also well recognized as a potential agent of bioterrorism. Routine culture and biochemical testing methods are useful for the identification of Bacillus anthracis, but a definitive identification may take 24 to 48 h or longer and may require that specimens be referred to another laboratory. Virulent isolates of B. anthracis contain two plasmids (pX01 and pX02) with unique targets that allow the rapid and specific identification of B. anthracis by PCR. We ...

  15. TRANSGENIC PLANTS EXPRESSING BACILLUS THURINGIENSIS DELTA-ENDOTOXINS

    Institute of Scientific and Technical Information of China (English)

    Hua-rong,Li; BrendaOppert; KunYanZhu; RandallA.Higgins; Fang-nengHuang; LawrentL.Buschman

    2003-01-01

    Commercial varieties of transgenic Bacillus thuringiensis (Bt) plants have been developed in many countries to control target pests. Initially, the expression of native Bt genes in plants was low due to mRNA instability, improper splicing, and post-translation modifications. Subsequently, modifications of the native Bt genes greatly enhanced expression levels. This is a review of the developments that made modem high-expression transgenic Bt plants possible, with an emphasis on the reasons for the low-level expression of native Bt genes in plant systems, and the techniques that have been used to improve plant expression of Bt toxin genes.

  16. Bacillus Calmette-Guérin vaccination at birth

    DEFF Research Database (Denmark)

    Kjærgaard, Jesper; Stensballe, Lone Graff; Birk, Nina Marie;

    2016-01-01

    BACKGROUND: Bacillus Calmette-Guérin vaccine (BCG) induces a complex, pro-inflammatory immune response. Obesity is associated with low-grade inflammation. AIMS: The purpose of the study was to test whether BCG at birth has effects on infant growth and body composition. STUDY DESIGN, SUBJECTS......-upper-arm circumference, or triceps and subscapular skinfold at 13months. CONCLUSION: In this study, vaccination with BCG at birth did not have effects on child growth or body composition at 13months. TRIAL REGISTRATION: www.clinicaltrials.gov, registration number NCT01694108....

  17. Ultra-violet-resistant mutants of Bacillus thuringiensis

    Energy Technology Data Exchange (ETDEWEB)

    Jones, D.R.; Karunakaran, V. (Polytechnic of Central London (UK). Faculty of Engineering and Science, School of Biological and Health Sciences); Burges, H.D. (Institute of Horticultural Research, Littlehampton (UK)); Hacking, A.J. (Reading Univ. (UK). Dextra Labs.Ltd.)

    1991-06-01

    One of the main disadvantages of using Bacillus thuringiensis as an insecticide is that the spore and crystal preparations applied to foliage are readily washed away by rain and are inactivated by sunlight. Spores from some strains of B. thuringiensis have been shown to be highly sensitive to u.v. light. This study has demonstrated how mutants with increased resistance to u.v., isolated by successive rounds of u.v. irradiation, and additionally with increased specific pathogenicity can be isolated. These techniques should be applied to strains that are frequently used in the industrial production of B.thuringiensis toxin. (author).

  18. Occurrence and diversity of mosquitocidal strains of Bacillus thuringiensis

    Directory of Open Access Journals (Sweden)

    K. Balaraman

    2005-09-01

    Full Text Available Ever since the discovery of the first Bacillus thuringiensis strain capable of killing mosquito larvae,namely, B. thuringiensis var israelensis, there are several reports from different parts of the worldabout the occurrence of mosquitocidal strains belonging to different subspecies/serotypes numberingthirty-six. The main sources of these wild type strains are soils/sediments, plants, animal feces,sick/moribund insects and waters. The toxicity of the strains within a subspecies/serotype variedwidely. Some of the strains exhibited toxicity to mosquitoes as well as lepidopterans and dipterans(including mosquitoes as well as plant parasitic nematodes.

  19. Bacillus thuringiensis resistance in Plutella - too many trees?

    Science.gov (United States)

    Crickmore, Neil

    2016-06-01

    Plutella xylostella was the first insect for which resistance to Bacillus thuringiensis was reported in the field, yet despite many studies on the nature of this resistance phenotype its genetic and molecular basis remains elusive. Many different factors have been proposed as contributing to resistance, although in many cases it has not been possible to establish a causal link. Indeed, there are so many studies published that it has become very difficult to 'see the wood for the trees'. This article will attempt to clarify our current understanding of Bt resistance in P. xylostella and consider the criteria that are used when validating a particular model.

  20. Gramicidin S production by Bacillus brevis in simulated microgravity

    Science.gov (United States)

    Fang, A.; Pierson, D. L.; Mishra, S. K.; Koenig, D. W.; Demain, A. L.

    1997-01-01

    In a continuing study of microbial secondary metabolism in simulated microgravity, we have examined gramicidin S (GS) production by Bacillus brevis strain Nagano in NASA High Aspect Rotating Vessels (HARVs), which are designed to simulate some aspects of microgravity. Growth and GS production were found to occur under simulated microgravity. When performance under simulated microgravity was compared with that under normal gravity conditions in the bioreactors, GS production was found to be unaffected by simulated microgravity. The repressive effect of glycerol in flask fermentations was not observed in the HARV. Thus the negative effect of glycerol on specific GS formation is dependent on shear and/or vessel geometry, not gravity.

  1. Ecotoxicity studies of antifungal metabolites of Bacillus sp. IBA 33.

    OpenAIRE

    MA Gordillo; RL Gomez; AR Navarro; MC Maldonado

    2015-01-01

    El incremento de enfermedade s de plantas causado por la proliferación de patógenos resis tentes a fungicidas intensificó la investigación de nuevos metabo litos activos contra ellos. El es tudio de métodos biológicos como una a lternativa al control químico h a alcanzado relevancia en años r ecientes. Se ha sugerido que el u so de metabolitos de Bacillus , es una alternativa o un método suplementario a la protección química de las plantas, siempre y cuan...

  2. [Pulmonal bacillus Calmette-Guérin infection two years after intravesical bacillus Calmette-Guérin installation].

    Science.gov (United States)

    Harbjerg, Julie Lykke; Bjerre, Cathrine Collin; Lillebæk, Troels; Weinreich, Ulla Møller

    2014-12-15

    Major complications following installation of bacillus Calmette Guérin (BCG) in the bladder as treatment for early stage cancer are rare and there are only few reports of BCG isolated from patients with suspected "BCGitis". We report a case where sputum culture yielded a BCG strain identical to one used two years earlier to treat the patient's bladder cancer. Some aspects were challenging in differential diagnosis: the patient's prior history of pulmonary tuberculosis ten years earlier and a two-year interval since BCG installation. PCR analysis and culture finally provided the diagnosis disseminated BCG infection.

  3. PRODUKSI ANTIBIOTIKA OLEH Bacillus subtilis M10 DALAM MEDIA UREA-SORBITOL

    Directory of Open Access Journals (Sweden)

    Supartono Supartono

    2012-04-01

    Full Text Available PRODUCTION OF ANTIBIOTICS BY Bacillus subtilis M10 IN UREA-SORBITOL MEDIUM. Infection diseases still become the main health problems that suffered by people in Indonesia. Besides, there were many pathogen bacteria found to be resistant to the some antibiotics. Therefore, the efforts to get a new antibiotic require to be done continuously. A new local strain of Bacillus subtilis BAC4 has been known producing an antibiotic that inhibit Serratia marcescens ATCC 27117 growth. To make efficient the local strain, mutation on Bacillus subtilis BAC4 was done by using acridine orange and a mutant cell of Bacillus subtilis M10 that overproduction for producing antibiotic was obtained. Nevertheless, the production kinetics of antibiotic by this mutant has not been reported. The objective of this research was to study the production kinetics of antibiotic by Bacillus subtilis M10 mutant. The production of antibiotic was conducted using batch fermentation and antibiotic assay was performed with agar absorption method using Serratia marcescens ATCC 27117 as bacteria assay. Research result provided that Bacillus subtilis M10 mutant with overproduction of antibiotic produced an antibiotic since 8th hour’s fermentation and optimum of it production was at 14th hours after inoculation.  Penyakit infeksi masih menjadi masalah yang utama diderita oleh masyarakat Indonesia. Di samping itu, banyak bakteri patogen yang ditemukan resisten terhadap beberapa antibiotika. Oleh karena itu, upaya-upaya untuk mendapatkan antibiotika baru perlu dilakukan secara terus-menerus. Suatu galur lokal baru Bacillus subtilis BAC4 teridentifikasi memproduksi senyawa antibiotika yang menghambat pertumbuhan Serratia marcescens ATCC27117. Untuk memberdayakan galur tersebut, terhadap Bacillus subtilis BAC4 dilakukan mutasi dengan larutan akridin oranye dan diperoleh mutan Bacillus subtilis M10 yang memproduksi antibiotika berlebihan. Namun, kinetika produksi antibiotika oleh Bacillus

  4. Transport of Bacillus thuringiensis var. kurstaki via fomites.

    Science.gov (United States)

    Van Cuyk, Sheila; Veal, Lee Ann B; Simpson, Beverley; Omberg, Kristin M

    2011-09-01

    The intentional and controlled release of an aerosolized bacterium provides an opportunity to investigate the implications of a biological attack. Since 2006, Los Alamos National Laboratory has worked with several urban areas, including Fairfax County, VA, to design experiments to evaluate biodefense concepts of operations using routine spraying of Bacillus thuringiensis var. kurstaki (Btk). Btk is dispersed in large quantities as a slurry to control the gypsy moth, Lymantria dispar. Understanding whether personnel and equipment pick up residual contamination during sampling activities and transport it to other areas is critical for the formulation of appropriate response and recovery plans. While there is a growing body of literature surrounding the transmission of viral diseases via fomites, there is limited information on the transport of Bacillus species via this route. In 2008, LANL investigated whether field sampling activities conducted near sprayed areas, post-spray, resulted in measurable cross-contamination of sampling personnel, equipment, vehicles, and hotel rooms. Viable Btk was detected in all sample types, indicating transport of the agent occurred via fomites.

  5. Methodology for fast evaluation of Bacillus thuringiensis crystal protein content

    Directory of Open Access Journals (Sweden)

    Alves Lúcia M. Carareto

    2000-01-01

    Full Text Available The development of the production and use of Bacillus thuringiensis in Brazil at a commercial scale faces certain difficulties, among them the establishment of efficient methodologies for the quantitation of toxic products to be commercialized. Presently, the amount of toxin is given in percentage by analyzing the samples total protein content. Such methodology however, does not measure the actual amount of active protein present in the product, since most strains express different endotoxin genes and might even produce b-toxin. Since the various types of toxins exhibit different antigenic characteristics, this work has as objective the utilization of fast immunological techniques to quantify the level of crystal protein. Crystal protein produced by a subspecies of Bacillus thuringiensis var. israelensis was purified by ultracentrifugation and utilized to immunize rabbits and to produce hiperimmune sera. Such sera were latter used to evaluate the level of proteins on commercial bioinsecticide and on laboratory cultures of B. thuringiensis through the immunodot technique. The results were obtained by comparison of data obtained from reactions with known concentrations of crystal protein permitting to evaluate the level of such protein on various materials.

  6. Plasmid-associated sensitivity of Bacillus thuringiensis to UV light

    International Nuclear Information System (INIS)

    Spores and vegetative cells of Bacillus thuringiensis were more sensitive to UV light than were spores or cells of plasmid-cured B. thuringiensis strains or of the closely related Bacillus cereus. Introduction of B. thuringiensis plasmids into B. cereus by cell mating increased the UV sensitivity of the cells and spores. Protoxins encoded by one or more B. thuringiensis plasmids were not involved in spore sensitivity, since a B. thuringiensis strain conditional for protoxin accumulation was equally sensitive at the permissive and nonpermissive temperatures. In addition, introduction of either a cloned protoxin gene, the cloning vector, or another plasmid not containing a protoxin gene into a plasmid-cured strain of B. thuringiensis all increased the UV sensitivity of the spores. Although the variety of small, acid-soluble proteins was the same in the spores of all strains examined, the quantity of dipicolinic acid was about twice as high in the plasmid-containing strains, and this may account for the differences in UV sensitivity of the spores. The cells of some strains harboring only B. thuringiensis plasmids were much more sensitive than cells of any of the other strains, and the differences were much greater than observed with spores

  7. Application of gaseous ozone for inactivation of Bacillus subtilis spores.

    Science.gov (United States)

    Aydogan, Ahmet; Gurol, Mirat D

    2006-02-01

    The effectiveness of gaseous ozone (O3) as a disinfectant was tested on Bacillus subtilis spores, which share the same physiological characteristics as Bacillus anthracis spores that cause the anthrax disease. Spores dried on surfaces of different carrier material were exposed to O3 gas in the range of 500-5000 ppm and at relative humidity (RH) of 70-95%. Gaseous O3 was found to be very effective against the B. subtilis spores, and at O3 concentrations as low as 3 mg/L (1500 ppm), approximately 3-log inactivation was obtained within 4 hr of exposure. The inactivation curves consisted of a short lag phase followed by an exponential decrease in the number of surviving spores. Prehydration of the bacterial spores has eliminated the initial lag phase. The inactivation rate increased with increasing O3 concentration but not >3 mg/L. The inactivation rate also increased with increase in RH. Different survival curves were obtained for various surfaces used to carry spores. Inactivation rates of spores on glass, a vinyl floor tile, and office paper were nearly the same. Whereas cut pile carpet and hardwood flooring surfaces resulted in much lower inactivation rates, another type of carpet (loop pile) showed significant enhancement in the inactivation of the spores. PMID:16568801

  8. Plasmid-associated sensitivity of Bacillus thuringiensis to UV light

    Energy Technology Data Exchange (ETDEWEB)

    Benoit, T.G.; Wilson, G.R.; Bull, D.L.; Aronson, A.I. (Department of Agriculture, College Station, TX (USA))

    1990-08-01

    Spores and vegetative cells of Bacillus thuringiensis were more sensitive to UV light than were spores or cells of plasmid-cured B. thuringiensis strains or of the closely related Bacillus cereus. Introduction of B. thuringiensis plasmids into B. cereus by cell mating increased the UV sensitivity of the cells and spores. Protoxins encoded by one or more B. thuringiensis plasmids were not involved in spore sensitivity, since a B. thuringiensis strain conditional for protoxin accumulation was equally sensitive at the permissive and nonpermissive temperatures. In addition, introduction of either a cloned protoxin gene, the cloning vector, or another plasmid not containing a protoxin gene into a plasmid-cured strain of B. thuringiensis all increased the UV sensitivity of the spores. Although the variety of small, acid-soluble proteins was the same in the spores of all strains examined, the quantity of dipicolinic acid was about twice as high in the plasmid-containing strains, and this may account for the differences in UV sensitivity of the spores. The cells of some strains harboring only B. thuringiensis plasmids were much more sensitive than cells of any of the other strains, and the differences were much greater than observed with spores.

  9. Report: antibiotic production by thermophilic Bacillus specie SAT-4.

    Science.gov (United States)

    Muhammad, Syed Aun; Ahmad, Safia; Hameed, Abdul

    2009-07-01

    Production of antimicrobial compounds seems to be a general phenomenon for most bacteria. The prevalence of antimicrobial resistance among key microbial pathogens is increasing at an alarming rate worldwide. Current solutions involve development of a more rationale approach to antibiotic use and discover of new antimicrobials. Bacillus species produce a large number of biological compounds active against bacteria, fungi, protozoa and viruses. The process of production usually involves screening of wide range of microorganisms, testing and modification. Production is carried out using fermentation. Thermophilic spore-forming, gram positive, motile rod bacterial strains were isolated from the Thar Desserts, Sindh Province, Pakistan. These strains were screened and checked for antibacterial activity. The best activity was observed by SAT4 against Micrococcus luteus, Staphylococcus aureus and Pseudomonas aeroginosa. The activity was only observed against gram positive bacteria and no activity was seen against Pseudomonas aeroginosa. Thermophilic Bacillus specie SAT4 was found to be active in the fermentation process to produce the antimicrobial agents. Further optimizations of different conditions (time of incubation, media, pH, glucose concentrations, nitrogen concentrations, and temperature) for antimicrobial production by the selected bacterial strain was performed. Agar diffusion assay was performed to evaluate the antibacterial activity. Optimum conditions for the production of antimicrobials by selected isolate were observed to be 48 hour, pH 5, temperature 55 degrees C, 2% glucose and 1.5% nitrogen concentration. This newly isolated bacterial strain has great potential for antimicrobial production at industrial scale. PMID:19553186

  10. Characteristics of bacillus strains with antifungal activity against phytopathogens

    International Nuclear Information System (INIS)

    Four bacterial isolates that showed antifungal activity against Alternaria alternata and other phytopathogens were isolates from bean rhizosphere. 16S rDNA analysis and phylogenetic relationship indicated that these isolates belong to Genus Bacillus. Isolate A1 clustered with Bacillus licheniformis while other isolates A2, A3 and A4 clustered together with B.pumilus. n-Butanol extract of these isolates strongly inhibited the growth of A. alternata while, chloroform extract of isolate A2 and ethyl acetate extract of A1,A3, and A4 inhibited the test fungus partially. All the isolates except A4 produced chitinase enzyme. None of the isolates solubilized mineral phosphate. Radiation sensitivity of isolates A1, A2, A3 and A4 were assessed and the LD99 values are determined as 0.50, 6.69, 11,60, 1.53 kGy, respectively. Mutant libraries of each isolate were prepared by exposing them to gamma radiation at their respective LD99 dose. Crude metabolite caused drastic changes on A. alternata hyphal morphology. Appearance of shrunken and collapsed hyphae could be due to the leak of cell wall or changes in membrane permeability

  11. Bacteriocins synthesized by Bacillus thuringiensis: generalities and potential applications

    Science.gov (United States)

    Salazar-Marroquín, Elma Laura; Galán-Wong, Luis J.; Moreno-Medina, Víctor Ricardo; Reyes-López, Miguel Ángel; Pereyra-Alférez, Benito

    2016-01-01

    The members of the Bacillus thuringiensis group, commonly known as Bt, produce a huge number of metabolites, which show biocidal and antagonistic activity. B. thuringiensis is widely known for synthesizing Cry, Vip and Cyt proteins, active against insects and other parasporins with biocidal activity against certain types of cancerous cells. Nevertheless, B. thuringiensis also synthesizes compounds with antimicrobial activity, especially bacteriocins. Some B. thuringiensis bacteriocins resemble lantibiotics and other small linear peptides (class IIa) from the lactic acid bacteria bacteriocins classification system. Although many bacteriocins produced by Bt have been reported, there is no proper classification for them. In this work, we have grouped these based on molecular weight and functionality. Bacteriocins are small peptides synthesized by bacteria, presenting inhibitory activity against Gram-positive and Gram-negative bacteria and to a lesser extent against fungi. These molecules represent a good study model in the search for microbial control alternatives. Lactic acid bacteria produces a huge number of these types of molecules with great potential. Nonetheless, members of the Bacillus, cereus group, especially B. thuringiensis, emerge as an attractive alternative for obtaining bacteriocins showing novel activities. This review describes the potential applications of B. thuringiensis bacteriocins in the control of foodborne pathogens, environment and medical area. PMID:27340340

  12. Bacillus amyloliquefaciens SUBSP. plantarum PROBIOTIC STRAINS AS PROTEASE PRODUCERS

    Directory of Open Access Journals (Sweden)

    E. V. Маtseliukh

    2015-04-01

    Full Text Available Proteases from probiotic strains of the genus Bacillus, just like the antibiotics, bacteriocins and other hydrolytic enzymes, are one of the main factors that determine their biological activity. The aim of this work was to study the synthesis and biochemical properties of proteases from two strains Bacillus amyloliquefaciens subsp. plantarum UCM B-5139 and UCM B-5140 that included in the probiotic Endosporin. The cultivation of strains was carried out in flasks under rotating for two days. The influence of physico-chemical parameters of the reaction medium on proteolytic activity was studied on partially purified protease preparations. Lytic activity was determined by turbidimetric method. On the second day of cultivation B. amyloliquefaciens subsp. plantarum UCM В-5139 and UCM В-5140 synthesized the metal-dependent peptidase and serine protease, respectively. The optimum conditions of their action were the following: temperature 37–40 °C and pH 6.5–7.0. Isolated proteases are able to lyse the living cells of Staphylococcus aureus and Candida albicans. Thus we demonstrated that B. amyloliquefaciens subsp. plantarum UCM B-5140 and UCM B-5139, included in the probiotic veterinary preparation Endosporin, produced proteolytic enzymes that hydrolyze the native insoluble proteins (elastin, fibrin and collagen. These enzymes belong to the group of neutral metal-dependent and serine proteases. They are active under physiological conditions against gram-positive bacteria and yeasts. The application of these proteases in biotechnology is considered.

  13. Isolation and Characterization of Phages Infecting Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Anna Krasowska

    2015-01-01

    Full Text Available Bacteriophages have been suggested as an alternative approach to reduce the amount of pathogens in various applications. Bacteriophages of various specificity and virulence were isolated as a means of controlling food-borne pathogens. We studied the interaction of bacteriophages with Bacillus species, which are very often persistent in industrial applications such as food production due to their antibiotic resistance and spore formation. A comparative study using electron microscopy, PFGE, and SDS-PAGE as well as determination of host range, pH and temperature resistance, adsorption rate, latent time, and phage burst size was performed on three phages of the Myoviridae family and one phage of the Siphoviridae family which infected Bacillus subtilis strains. The phages are morphologically different and characterized by icosahedral heads and contractile (SIOΦ, SUBω, and SPOσ phages or noncontractile (ARπ phage tails. The genomes of SIOΦ and SUBω are composed of 154 kb. The capsid of SIOΦ is composed of four proteins. Bacteriophages SPOσ and ARπ have genome sizes of 25 kbp and 40 kbp, respectively. Both phages as well as SUBω phage have 14 proteins in their capsids. Phages SIOΦ and SPOσ are resistant to high temperatures and to the acid (4.0 and alkaline (9.0 and 10.0 pH.

  14. Efficiency of Intergeneric Recombinants Between Bacillus Thuringiensis and Bacillus Subtilis for Increasing Mortality Rate in Cotten Leaf Worm

    Science.gov (United States)

    AlOtaibi, Saad Aied

    2012-12-01

    In this study , two strains of Bacillus belonging to two serotypes and four of their transconjugants were screened with respect to their toxicity against lepidopterous cotton pest. . Bacterial transconjugants isolated from conjugation between both strains were evaluated for their transconjugant efficiency caused mortality in Spodoptera littoralis larvae . Two groups of bioinsecticides ; crystals , crystals and spores have been isolated from Bacillusstrains and their transconjugants . Insecticidal crystal protein ( ICP ) was specific for lepidopteran insects because of the toxin sufficient both for insect specificity and toxicity . The toxicities of these two groups against larvae of Spodoptera littoralis was expressed as transconjugant efficiency , which related to the mean number of larvae died expressed as mortality percentage . The results showed transconjugant efficiency in reducing the mean number of Spodoptera littoralis larvae feeding on leaves of Ricinus communis sprayed with bioinsecticides of Bt transconjugants. Most values of positive transconjugant efficiency related to increasing mortality percentage are due to toxicological effects appeared in response to the treatments with crystals + endospores than that of crystals alone .This indicated that crystals + endospores was more effective for increasing mortality percentage than that resulted by crystals . Higher positive transconjugant efficiency in relation to the mid parents and better parent was appeared at 168 h of treatment . The results indicated that recombinant Bacillus thuringiensis are important control agents for lepidopteran pests , as well as , susceptibility decreased with larval development . The results also suggested a potential for the deployment of these recominant entomopathogens in the management of Spodoptera. littoralis larvae .

  15. Isolation and Identification of the Antimicrobial Substance Produced by Bacillus subtilis fmbR%Bacillus subtilis fmbR抗菌物质的分离和鉴定

    Institute of Scientific and Technical Information of China (English)

    别小妹; 陆兆新; 吕凤霞; 赵海珍; 杨胜远; 孙力军

    2006-01-01

    [目的]对Bacillus subtilis fmbR产生的抗菌物质进行分离和鉴定研究,以确定抗菌物质的组成和结构.[方法]采用HPLC和TLC层析对Bacillus subtilis fmbR抗菌物质进行分离纯化,通过ESI-MS和MALDI-MS分析对抗菌物质的组成和结构进行初步鉴定.[结果]HPLC层析表明了Bacillus subtilis fmbR抗菌物质含有保留时间与surfactin相似的成分.TLC层析和原位酸解证明了Bacillus subtilis fmbR抗菌物质含有闭合肽键类的物质,其中之一为相对迁移率Rf与标样surfactin相近的组分.采用ESI-MS分析检测到Bacillus subtilis fmbR抗菌物质含有分子量与surfactinA相同的m/z1009.1、m/z1023.2 和m/z1037.0等3种同系物;通过MALDI-MS分析获得[M+H]+为m/z 3403.95抗菌物质,该物质分子量与Bacillus subtilis 168产生的细菌素subtilosin的m/z3403.3 相同.[结论]Bacillus subtilis fmbR抗菌物质由C13~C15的3种surfactinA同系物和一种羊毛硫抗生素subtilosin组成.

  16. Diversity of Secondary Metabolites from Marine Bacillus Species: Chemistry and Biological Activity

    Directory of Open Access Journals (Sweden)

    Hee Jae Shin

    2013-08-01

    Full Text Available Marine Bacillus species produce versatile secondary metabolites including lipopeptides, polypeptides, macrolactones, fatty acids, polyketides, and isocoumarins. These structurally diverse compounds exhibit a wide range of biological activities, such as antimicrobial, anticancer, and antialgal activities. Some marine Bacillus strains can detoxify heavy metals through reduction processes and have the ability to produce carotenoids. The present article reviews the chemistry and biological activities of secondary metabolites from marine isolates. Side by side, the potential for application of these novel natural products from marine Bacillus strains as drugs, pesticides, carotenoids, and tools for the bioremediation of heavy metal toxicity are also discussed.

  17. Sensitivity of the bacterium Bacillus Thuringiensis as an insect disease agent to gamma-rays

    International Nuclear Information System (INIS)

    The effect of gamma radiation on the viability of the entomopathogenic spore-forming bacterium, Bacillus thuringiensis, was tested. The different gamma doses varied much in their effect on such bacterium. All irradiated Bacillus suspensions with doses below 85 krad showed different degrees of inhibitory activity. However, bacterial suspensions irradiated at a dose of 90 krad. proved to promote spore germination. Changes in the physiological, and morphological characters of the irradiated Bacillus at these levels were detected. The new observed characters were induced at a particular dose level of 90 krad. These new characters are assumed to be due to genetic changes induced at this particular gamma dose

  18. Isolation and characterization of antagonistic Bacillus strains capable to degrade ethylenethiourea.

    Science.gov (United States)

    Vágvölgyi, Csaba; Sajben-Nagy, Enikő; Bóka, Bettina; Vörös, Mónika; Berki, Adrienn; Palágyi, Andrea; Krisch, Judit; Skrbić, Biljana; Durišić-Mladenović, N; Manczinger, László

    2013-03-01

    In this study, more than 150 bacteria showing antagonistic properties against bacterial and fungal pathogens of the tomato plant were isolated and characterized. The most efficient agents against these phytopathogenic microorganisms belong to the genus Bacillus: the best biocontrol isolates were representatives of Bacillus subtilis, B. mojavensis and B. amyloliquefaciens species. They intensively produced fengycin or/and surfactin depsipeptide antibiotics and also proved to be excellent protease secretors. It was proved, that the selected strains were able to use ethylenethiourea (ETU) as sole nitrogen source. These antagonistic and ETU-degrading Bacillus strains can be applied as biocontrol and also as bioremediation agents. PMID:23143288

  19. Activation of the latent PlcR regulon in Bacillus anthracis

    OpenAIRE

    Sastalla, Inka; Maltese, Lauren M.; Pomerantseva, Olga M.; Pomerantsev, Andrei P; Keane-Myers, Andrea; Stephen H Leppla

    2010-01-01

    Many genes in Bacillus cereus and Bacillus thuringiensis are under the control of the transcriptional regulator PlcR and its regulatory peptide, PapR. In Bacillus anthracis, the causative agent of anthrax, PlcR is inactivated by truncation, and consequently genes having PlcR binding sites are expressed at very low levels when compared with B. cereus. We found that activation of the PlcR regulon in B. anthracis by expression of a PlcR–PapR fusion protein does not alter sporulation in strains c...

  20. Suitable conditions for xylanases activities from Bacillus sp. GA2(1 and Bacillus sp. GA1(6 and their properties for agricultural residues hydrolysis

    Directory of Open Access Journals (Sweden)

    Sudathip Chantorn

    2016-04-01

    Full Text Available Bacillus sp. GA2(1 and Bacillus sp. GA1(6 were isolated from soybean field in Khon Kaen province, Thailand. Crude enzymes from both isolates showed the activities of cellulase, xylanase, and mannanase at 37°C for 24 h. The highest xylanase activities of Bacillus sp. GA2(1 and Bacillus sp. GA1(6 were 1.58±0.25 and 0.82±0.16 U/ml, respectively. The relative xylanase activities from both strains were more than 60% at pH 5.0 to 8.0. The optimum temperature of xylanases was 50°C in both strains. The residual xylanase activities from both strains were more than 70% at 60°C for 60 min. Five agricultural wastes (AWs, namely coffee residue, soybean meal, potato peel, sugarcane bagasse, and corn cobs, were used as substrates for hydrolysis properties. The highest reducing sugar content of 101±1.32 µg/ml was obtained from soybean meal hydrolysate produced by Bacillus sp. GA2(1 xylanase.

  1. Genotyping of starter cultures of Bacillus subtilis and Bacillus pumilus for fermentation of African locust bean (Parkia biglobosa) to produce Soumbala.

    Science.gov (United States)

    Ouoba, Labia Irène Ivette; Diawara, Bréhima; Amoa-Awua, Wisdom kofi; Traoré, Alfred Sababénedyo; Møller, Peter Lange

    2004-01-15

    Bacillus spp. are the predominant microorganisms in fermented African locust bean called Soumbala in Burkina Faso. Ten strains selected as potential starter cultures were characterised by PCR amplification of the16S-23S rDNA intergenic transcribed spacer (ITS-PCR), restriction fragment length polymorphism of the ITS-PCR (ITS-PCR RFLP), pulsed field gel electrophoresis (PFGE) and sequencing of the 968-1401 region of the 16S rDNA. In previous studies, the isolates were identified by phenotyping as Bacillus subtilis and Bacillus pumilus. The phenotyping was repeated as a reference in the present study. The ITS-PCR and ITS-PCR RLFP allowed a typing at species level. The PFGE was more discriminative and allowed a typing at strain level. Full agreement with the phenotyping was observed in all cases. The sequencing of the 16S rDNA allowed the identification at species level with an identity from 97% to 100% comparing the sequences to those from the GenBank databases. The desired cultures of B. subtilis and B. pumilus from African locust bean fermentation were distinguished by ITS-PCR and ITS-PCR RLFP from Bacillus cereus and Bacillus sphaericus which sometimes occur in the beginning of the fermentation.

  2. Bacillus anthracis Factors for Phagosomal Escape

    Directory of Open Access Journals (Sweden)

    Irene Zornetta

    2012-07-01

    Full Text Available The mechanism of phagosome escape by intracellular pathogens is an important step in the infectious cycle. During the establishment of anthrax, Bacillus anthracis undergoes a transient intracellular phase in which spores are engulfed by local phagocytes. Spores germinate inside phagosomes and grow to vegetative bacilli, which emerge from their resident intracellular compartments, replicate and eventually exit from the plasma membrane. During germination, B. anthracis secretes multiple factors that can help its resistance to the phagocytes. Here the possible role of B. anthracis toxins, phospholipases, antioxidant enzymes and capsules in the phagosomal escape and survival, is analyzed and compared with that of factors of other microbial pathogens involved in the same type of process.

  3. Cloning and Characterization of Gene Promoters from Bacillus pumilus

    Institute of Scientific and Technical Information of China (English)

    Pan Jiao(潘皎); Zhang Yizheng

    2004-01-01

    DNA fragments obtained from Sau3AI partially digested total DNA of Bacillus pumilus UN31-C-42 are first inserted into BamHI site of pSUPV4, a promoter-probe vector. The recombinant DNA molecules are transformed into Escherichia coli cells and eight-three Kanr clones (named pSUBp1- pSUBp83) are obtained. The inserted fragments in pSUBp53, pSUBp57, pSUBp21, which showed high level of kanamycin - resistance, are sequenced and analyzed, respectively. These fragments contain some conserved sequences of prokaryotic gene promoters, such as TATAAT and TTGACA box. The promoter fragment Bp53 could efficiently promote the alkaline protease gene of B.pumilus expression not only in E.coli but also in B.subtilis cells.

  4. A new alkaline elastase of an alkalophilic bacillus.

    Science.gov (United States)

    Tsai, Y C; Yamasaki, M; Yamamoto-Suzuki, Y; Tamura, G

    1983-11-01

    A new alkaline elastase was purified from the culture broth of an alkalophilic Bacillus sp. Ya-B. This was a serine proteinase. Molecular weight was 25,000. The optimum pH for elastin and casein was 11.75. The enzyme had very high specific activity, 12,400 units/mg protein for casein, and 2,440 units/mg protein for elastin at the optimum pH. It showed marked preference for elastin. The relative activity of elastin/casein of this enzyme was 17 and 6 times higher than those of subtilisin BPN' and subtilisin Carlsberg, respectively. This enzyme also had higher keratin and collagen hydrolyzing activity in comparison with subtilisin.

  5. Genetics of thermophilic bacteria. [Bacillus stearothermophilus:a2

    Energy Technology Data Exchange (ETDEWEB)

    Welker, N.E.

    1991-01-01

    Organisms adapted to high temperature have evolved a variety of unique solutions to the biochemical problems imposed by this environment. Adaptation is commonly used to describe the biochemical properties of organisms which have become adapted to their environment (genetic adaptation). It can also mean the direct response-at the cellular level-of an organism to changes in temperature (physiological adaptation). Thermophilic bacilli (strains of Bacillus stearothermophilus) can exhibit a variety of biochemical adaptations in response to changes in temperature. These include changes in the composition and stability of the membrane, metabolic potential, the transport of amino acids, regulatory mechanisms, ribose methylation of tRNA, protein thermostability, and nutritional requirements. The objectives of the research were to develop efficient and reliable genetic systems to analyze and manipulate B. Stearothermophilus, and to use these systems initiate a biochemical, molecular, and genetic investigations of genes that are required for growth at high temperature.

  6. Quorum Quenching Bacillus sonorensis Isolated from Soya Sauce Fermentation Brine

    Directory of Open Access Journals (Sweden)

    Kok-Gan Chan

    2012-03-01

    Full Text Available An N-acylhomoserine lactone (AHL-degrading bacterial strain, L62, was isolated from a sample of fermentation brine of Chinese soya sauce by using rich medium agar supplemented with soya sauce (10% v/v. L62, a rod-shaped Gram positive bacterium with amylolytic activity, was phylogentically related to Bacillus sonorensis by 16S ribosomal DNA and rpoB sequence analyses. B. sonorensis L62 efficiently degraded N-3-oxohexanoyl homoserine lactone and N-octanoylhomoserine lactone. However, the aiiA homologue, encoding an autoinducer inactivation enzyme catalyzing the degradation of AHLs, was not detected in L62, suggesting the presence of a different AHL-degrading gene in L62. To the best of our knowledge, this is the first report of AHL-degrading B. sonorensis from soya sauce liquid state fermentation.

  7. The respiratory arsenate reductase from Bacillus selenitireducens strain MLS10

    Science.gov (United States)

    Afkar, E.; Lisak, J.; Saltikov, C.; Basu, P.; Oremland, R.S.; Stolz, J.F.

    2003-01-01

    The respiratory arsenate reductase from the Gram-positive, haloalkaliphile, Bacillus selenitireducens strain MLS10 was purified and characterized. It is a membrane bound heterodimer (150 kDa) composed of two subunits ArrA (110 kDa) and ArrB (34 kDa), with an apparent Km for arsenate of 34 ??M and Vmax of 2.5 ??mol min-1 mg-1. Optimal activity occurred at pH 9.5 and 150 g l-1 of NaCl. Metal analysis (inductively coupled plasma mass spectrometry) of the holoenzyme and sequence analysis of the catalytic subunit (ArrA; the gene for which was cloned and sequenced) indicate it is a member of the DMSO reductase family of molybdoproteins. ?? 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

  8. Bacillus subtilis Hfq: A role in chemotaxis and motility

    Indian Academy of Sciences (India)

    CHANDRAKANT B JAGTAP; PRADEEP KUMAR; K KRISHNAMURTHY RAO

    2016-09-01

    Hfq is a global post-transcriptional regulator that modulates the translation and stability of target mRNAs and therebyregulates pleiotropic functions, such as growth, stress, virulence and motility, in many Gram-negative bacteria.However, comparatively little is known about the regulation and function(s) of Hfq in Gram-positive bacteria.Recently, in Bacillus subtilis, a role for Hfq in stationary phase survival has been suggested, although the possibilityof Hfq having an additional role(s) cannot be ruled out. In this study we show that an ortholog of Hfq in B. subtilis isregulated by the stress sigma factor, σB, in addition to the stationary phase sigma factor, σH. We further demonstratethat Hfq positively regulates the expression of flagellum and chemotaxis genes (fla/che) that control chemotaxis andmotility, thus assigning a new function for Hfq in B. subtilis.

  9. Nanoactivator mediated modifications in thermostable amylase from Bacillus licheniformis.

    Science.gov (United States)

    Khairnar, Rajendra S; Mahabole, Megha P; Pathak, Anupama P

    2012-12-01

    Gram-positive rod-shaped thermophilic bacteria were isolated using samples collected from terrestrial natural thermal spring located at Unkeshwar (Longitude 78.22 degree East to 78.34 degree East, Latitude 19 degree 34' North to 19 degree 40' North), District Nanded, Maharashtra State, India. The isolates were then cultivated using selective media and identified using culture-dependent techniques. One prominent isolate (UN1) exhibited high temperature stability and remarkable amylase production and was identified as Bacillus licheniformis. Amylase production was carried out in starch media and the enzyme was partially purified and characterized for optimization of pH and temperature. Amylolytic activity of the enzyme was determined. Nanoactivator-mediated modifications were carried out to enhance amylolytic activity of the partially purified amylase. Three-fold increase in catalytic efficiency of amylase was obtained after modification. PMID:23350283

  10. Stoichiometric growth model for riboflavin-producing Bacillus subtilis.

    Science.gov (United States)

    Dauner, M; Sauer, U

    2001-09-01

    Rate equations for measured extracellular rates and macromolecular composition data were combined with a stoichiometric model to describe riboflavin production with an industrial Bacillus subtilis strain using errors in variables regression analysis. On the basis of this combined stoichiometric growth model, we explored the topological features of the B. subtilis metabolic reaction network that was assembled from a large amount of literature. More specifically, we simulated maximum theoretical yields of biomass and riboflavin, including the associated flux regimes. Based on the developed model, the importance of experimental data on building block requirements for maximum yield and flux calculations were investigated. These analyses clearly show that verification of macromolecular composition data is important for optimum flux calculations. PMID:11505383

  11. Engineering of Bacillus subtilis 168 for increased nisin resistance

    DEFF Research Database (Denmark)

    Hansen, Mette; Wangari, Romilda; Hansen, Egon Bech;

    2009-01-01

    Nisin is a natural bacteriocin produced commercially by Lactococcus lactis and widely used in the food industry as a preservative because of its broad host spectrum. Despite the low productivity and troublesome fermentation of L. lactis, no alternative cost-effective host has yet been found....... Bacillus subtilis had been suggested as a potential host for the biosynthesis of nisin but was discarded due to its sensitivity to the lethal action of nisin. In this study, we have reevaluated the potential of B. subtilis as a host organism for the heterologous production of nisin. We applied...... transcriptome and proteome analyses of B. subtilis and identified eight genes upregulated in the presence of nisin. We demonstrated that the overexpression of some of these genes boosts the natural defenses of B. subtilis, which allows it to sustain higher levels of nisin in the medium. We also attempted...

  12. Bacillus anthracis secretes proteins that mediate heme acquisition from hemoglobin.

    Directory of Open Access Journals (Sweden)

    Anthony W Maresso

    Full Text Available Acquisition of iron is necessary for the replication of nearly all bacterial pathogens; however, iron of vertebrate hosts is mostly sequestered by heme and bound to hemoglobin within red blood cells. In Bacillus anthracis, the spore-forming agent of anthrax, the mechanisms of iron scavenging from hemoglobin are unknown. We report here that B. anthracis secretes IsdX1 and IsdX2, two NEAT domain proteins, to remove heme from hemoglobin, thereby retrieving iron for bacterial growth. Unlike other Gram-positive bacteria, which rely on cell wall anchored Isd proteins for heme scavenging, B. anthracis seems to have also evolved NEAT domain proteins in the extracellular milieu and in the bacterial envelope to provide for the passage of heme.

  13. Bacillus subtilis Hfq: A role in chemotaxis and motility.

    Science.gov (United States)

    Jagtap, Chandrakant B; Kumar, Pradeep; Rao, Krishnamurthy K

    2016-09-01

    Hfq is a global post-transcriptional regulator that modulates the translation and stability of target mRNAs and thereby regulates pleiotropic functions, such as growth, stress, virulence and motility, in many Gram-negative bacteria. However, comparatively little is known about the regulation and function(s) of Hfq in Gram-positive bacteria. Recently, in Bacillus subtilis, a role for Hfq in stationary phase survival has been suggested, although the possibility of Hfq having an additional role(s) cannot be ruled out. In this study we show that an ortholog of Hfq in B. subtilis is regulated by the stress sigma factor, sigma^B, in addition to the stationary phase sigma factor, sigma^H. We further demonstrate that Hfq positively regulates the expression of flagellum and chemotaxis genes (fla/che) that control chemotaxis and motility, thus assigning a new function for Hfq in B. subtilis. PMID:27581927

  14. Historical distribution and molecular diversity of Bacillus anthracis, Kazakhstan.

    Science.gov (United States)

    Aikembayev, Alim M; Lukhnova, Larissa; Temiraliyeva, Gulnara; Meka-Mechenko, Tatyana; Pazylov, Yerlan; Zakaryan, Sarkis; Denissov, Georgiy; Easterday, W Ryan; Van Ert, Matthew N; Keim, Paul; Francesconi, Stephen C; Blackburn, Jason K; Hugh-Jones, Martin; Hadfield, Ted

    2010-05-01

    To map the distribution of anthrax outbreaks and strain subtypes in Kazakhstan during 1937-2005, we combined geographic information system technology and genetic analysis by using archived cultures and data. Biochemical and genetic tests confirmed the identity of 93 archived cultures in the Kazakhstan National Culture Collection as Bacillus anthracis. Multilocus variable number tandem repeat analysis genotyping identified 12 genotypes. Cluster analysis comparing these genotypes with previously published genotypes indicated that most (n = 78) isolates belonged to the previously described A1.a genetic cluster, 6 isolates belonged to the A3.b cluster, and 2 belonged to the A4 cluster. Two genotypes in the collection appeared to represent novel genetic sublineages; 1 of these isolates was from Krygystan. Our data provide a description of the historical, geographic, and genetic diversity of B. anthracis in this Central Asian region. PMID:20409368

  15. Mode of action of mosquitocidal Bacillus thuringiensis toxins.

    Science.gov (United States)

    Soberón, Mario; Fernández, Luisa E; Pérez, Claudia; Gill, Sarjeet S; Bravo, Alejandra

    2007-04-01

    Cry toxins from Bacillus thuringiensis (Bt) are used for insect control. Their primary action is to lyse midgut epithelial cells. In lepidopteran insects, Cry1A monomeric toxins interact with a first receptor and this interaction triggers toxin oligomerization. The oligomeric structure interacts then with a second GPI-anchored receptor that induces insertion into membrane microdomains and larvae death. In the case of mosquitocidal Bt strains, two different toxins participate, Cry and Cyt. These toxins have a synergistic effect and Cyt1Aa overcomes Cry toxin-resistance. We will summarize recent findings on the identification of Cry receptors in mosquitoes and the mechanism of synergism: Cyt1Aa synergizes or suppresses resistance to Cry toxins by functioning as a Cry membrane-bound receptor. PMID:17145072

  16. Flexibility Analysis of Bacillus thuringiensis Cry1Aa

    Institute of Scientific and Technical Information of China (English)

    ZHAO Xin Min; XIA Li Qiu; YANG Xiao Ping; PENG Xiao Yun

    2015-01-01

    Objective To investigate the flexibility and mobility of the Bacillus thuringiensis toxin Cry1Aa. Methods The graph theory-based program Constraint Network Analysis and normal mode-based program NMsim were used to analyze the global and local flexibility indices as well as the fluctuation of individual residues in detail. Results The decrease in Cry1Aa network rigidity with the increase of temperature was evident. Two phase transition points in which the Cry1Aa structure lost rigidity during the thermal simulation were identified. Two rigid clusters were found in domains I and II. Weak spots were found in C-terminal domain III. Several flexible regions were found in all three domains;the largest residue fluctuation was present in the apical loop2 of domain II. Conclusion Although several flexible regions could be found in all the three domains, the most flexible regions were in the apical loops of domain II.

  17. Statistical analysis of cellulase production in Bacillus amyloliquefaciens UNPDV-22

    Directory of Open Access Journals (Sweden)

    Vasudeo Zambare

    2011-06-01

    Full Text Available The production of cellulase in Bacillus amyloliquefaciens UNPDV-22 was optimized usingresponse surface methodology (RSM. Central composite design (CCD was used to study the interactiveeffect of fermentation medium components (wheat bran, soybean meal, and malt dextrin on cellulaseactivity. Results suggested that wheat bran, soybean meal, and malt dextrin all have significant impacton cellulase production. The use of RSM resulted in a 70% increase in the cellulase activity over thecontrol of non-optimized basal medium. Optimum cellulase production of 11.23 U/mL was obtained in afermentation medium containing wheat bran (1.03%, w/v, soybean meal (2.43%, w/v, and maltdextrin (2.95%, w/v.

  18. Biosynthesis of isotopically labeled gramicidins and tyrocidins by Bacillus brevis

    Energy Technology Data Exchange (ETDEWEB)

    Vogt, T.C. Bas; Schinzel, Susan [Max-Planck-Institut fuer Biochemie (Germany); Bechinger, Burkhard [Universite Strasbourg, Faculte de chimie, Institut Le Bel (France)], E-mail: bechinger@chimie.u-strasbg.fr

    2003-05-15

    The three-dimensional structure of bilayer-associated gramicidin A is available from a structural data base. This and related peptides are, therefore, ideal model compounds to use during the implementation and development of new NMR techniques for the structural investigations of membrane proteins. As these methods rely on the isotopic labelling of single, selected or all sites, we have, investigated and optimised biochemical protocols using different strains of the Gram-positive bacterium Bacillus brevis. With newly developed schemes for isotopic labelling large amounts of gramicidin and tyrocidin enriched with stable isotopes such as {sup 15}N or {sup 15}N/{sup 13}C have been obtained at low cost. A variety of analytical and spectroscopic techniques, including HPLC, mass spectrometry and NMR spectroscopy are used to characterise the resulting products.

  19. Regulation of the anaerobic metabolism in Bacillus subtilis.

    Science.gov (United States)

    Härtig, Elisabeth; Jahn, Dieter

    2012-01-01

    The Gram-positive soil bacterium Bacillus subtilis encounters changing environmental conditions in its habitat. The access to oxygen determines the mode of energy generation. A complex regulatory network is employed to switch from oxygen respiration to nitrate respiration and various fermentative processes. During adaptation, oxygen depletion is sensed by the [4Fe-4S](2+) cluster containing Fnr and the two-component regulatory system ResDE consisting of the membrane-bound histidine kinase ResE and the cytoplasmic ResD regulator. Nitric oxide is the signal recognized by NsrR. Acetate formation and decreasing pH are measured via AlsR. Finally, Rex is responding to changes in the cellular NAD(+)/NADH ration. The fine-tuned interplay of these regulators at approximately 400 target gene promoters ensures efficient adaptation of the B. subtilis physiology. PMID:23046954

  20. Decontamination Options for Drinking Water Contaminated with Bacillus anthracis Spores

    Energy Technology Data Exchange (ETDEWEB)

    Raber, E; Burklund, A

    2010-02-16

    Five parameters were evaluated with surrogates of Bacillus anthracis spores to determine effective decontamination options for use in a contaminated drinking water supply. The parameters were: (1) type of Bacillus spore surrogate (B. thuringiensis or B. atrophaeus); (2) spore concentration in suspension (10{sup 2} to 10{sup 6} spores/ml); (3) chemical characteristics of decontaminant [sodium dicholor-s-triazinetrione dihydrate (Dichlor), hydrogen peroxide, potassium peroxymonosulfate (Oxone), sodium hypochlorite, and VirkonS{reg_sign}]; (4) decontaminant concentration (0.01% to 5%); and (5) decontaminant exposure time (10 min to 24 hr). Results from 162 suspension tests with appropriate controls are reported. Hydrogen peroxide at a concentration of 5%, and Dichlor and sodium hypochlorite at a concentration of 2%, were effective at spore inactivation regardless of spore type tested, spore exposure time, or spore concentration evaluated. This is the first reported study of Dichlor as an effective decontaminant for B. anthracis spore surrogates. Dichlor's desirable characteristics of high oxidation potential, high level of free chlorine, and more neutral pH than that of other oxidizers evaluated appear to make it an excellent alternative. All three oxidizers were effective against B. atrophaeus spores in meeting EPA's biocide standard of greater than a 6 log kill after a 10-minute exposure time and at lower concentrations than typically reported for biocide use. Solutions of 5% VirkonS{reg_sign} and Oxone were less effective decontaminants than other options evaluated in this study and did not meet the EPA's efficacy standard for biocides. Differences in methods and procedures reported by other investigators make quantitative comparisons among studies difficult.

  1. Nanoscale imaging of Bacillus thuringiensis flagella using atomic force microscopy

    Science.gov (United States)

    Gillis, Annika; Dupres, Vincent; Delestrait, Guillaume; Mahillon, Jacques; Dufrêne, Yves F.

    2012-02-01

    Because bacterial flagella play essential roles in various processes (motility, adhesion, host interactions, secretion), studying their expression in relation to function is an important challenge. Here, we use atomic force microscopy (AFM) to gain insight into the nanoscale surface properties of two wild-type and four mutant strains of Bacillus thuringiensis exhibiting various levels of flagellation. We show that, unlike AFM in liquid, AFM in air is a simple and reliable approach to observe the morphological details of the bacteria, and to quantify the density and dimensions of their flagella. We found that the amount of flagella expressed by the six strains, as observed at the nanoscale, correlates with their microscopic swarming motility. These observations provide novel information on flagella expression in Gram-positive bacteria and demonstrate the power of AFM in genetic studies for the fast assessment of the phenotypic characteristics of bacterial strains altered in cell surface appendages.Because bacterial flagella play essential roles in various processes (motility, adhesion, host interactions, secretion), studying their expression in relation to function is an important challenge. Here, we use atomic force microscopy (AFM) to gain insight into the nanoscale surface properties of two wild-type and four mutant strains of Bacillus thuringiensis exhibiting various levels of flagellation. We show that, unlike AFM in liquid, AFM in air is a simple and reliable approach to observe the morphological details of the bacteria, and to quantify the density and dimensions of their flagella. We found that the amount of flagella expressed by the six strains, as observed at the nanoscale, correlates with their microscopic swarming motility. These observations provide novel information on flagella expression in Gram-positive bacteria and demonstrate the power of AFM in genetic studies for the fast assessment of the phenotypic characteristics of bacterial strains altered in

  2. ENHANCED DEGRADATION OF CAPTAN BY IMMOBILIZED CELLS OF BACILLUS CIRCULANS

    Directory of Open Access Journals (Sweden)

    Veena More

    2014-10-01

    Full Text Available The possibility of using Bacillus circulans in degrading captan was evaluated by comparing the captan degradation rate by freely suspended and immobilized cells on agar, sodium alginate (SA, polyacrylamide (PA and polyurethane-foam (PUF in batch and repeated batch degradations. Under batch degradations, 50, 60, 72, and 88% of 0.1% captan was degraded by freely suspended cells, agar-, SA-, and PA-immobilized cells, respectively in 72 h; whereas 15, 47.5, 67.7 and 75% of 0.2% captan was degraded by freely suspended cells, agar-, SA-, and PA-immobilized cells, respectively in 72 h. However, 0.1 and 0.2% captan were completely degraded by PUF-immobilized cells in 48 and 72 h, respectively. Under repeated batch degradations, PUF-immobilized cells were reused more than 40 cycles for 72 h without losing the captan degradation ability, while the cells immobilized on agar, SA, and the PA could be reused for 15, 20, and 25 cycles, respectively. A significant 0.1% captan degradation by PUF-immobilized cells was observed at pH 4.0 - 10.0 and 20 - 40 ºC ranges. In contrast, freely suspended cells only degraded captan at optimum pH of 7.0 and 30 ºC. The PUF-immobilized cells were able to significantly degrade captan for 120 days at 4 ºC without losing the captan degradation ability; whereas this ability was lost in 120 days for freely suspended cells. Since the application of captan leads to pollution and reduces soil fertility, the use of immobilized cells of Bacillus circulans can thus be a better cost-effective strategy to decontaminate captan polluted sites.

  3. A Newly Isolated Thermostable Lipase from Bacillus sp.

    Directory of Open Access Journals (Sweden)

    Abu Bakar Salleh

    2011-05-01

    Full Text Available A thermophilic lipolytic bacterium identified as Bacillus sp. L2 via 16S rDNA was previously isolated from a hot spring in Perak, Malaysia. Bacillus sp. L2 was confirmed to be in Group 5 of bacterial classification, a phylogenically and phenotypically coherent group of thermophilic bacilli displaying very high similarity among their 16S rRNA sequences (98.5–99.2%. Polymerase chain reaction (PCR cloning of L2 lipase gene was conducted by using five different primers. Sequence analysis of the L2 lipase gene revealed an open reading frame (ORF of 1251 bp that codes for 417 amino acids. The signal peptides consist of 28 amino acids. The mature protein is made of 388 amino acid residues. Recombinant lipase was successfully overexpressed with a 178-fold increase in activity compared to crude native L2 lipase. The recombinant L2 lipase (43.2 kDa was purified to homogeneity in a single chromatography step. The purified lipase was found to be reactive at a temperature range of 55–80 °C and at a pH of 6–10. The L2 lipase had a melting temperature (Tm of 59.04 °C when analyzed by circular dichroism (CD spectroscopy studies. The optimum activity was found to be at 70 °C and pH 9. Lipase L2 was strongly inhibited by ethylenediaminetetraacetic acid (EDTA (100%, whereas phenylmethylsulfonyl fluoride (PMSF, pepstatin-A, 2-mercaptoethanol and dithiothreitol (DTT inhibited the enzyme by over 40%. The CD spectra of secondary structure analysis showed that the L2 lipase structure contained 38.6% α-helices, 2.2% ß-strands, 23.6% turns and 35.6% random conformations.

  4. Catalase activity as a biomarker for mild-stress-induced robustness in Bacillus weihenstephanensis

    NARCIS (Netherlands)

    Besten, den H.M.W.; Effraimidou, S.; Abee, T.

    2013-01-01

    Microorganisms are able to survive and grow in changing environments by activating stress adaptation mechanisms which may enhance bacterial robustness. Stress-induced enhanced robustness complicates the predictability of microbial inactivation. Using psychrotolerant Bacillus weihenstephanensis strai

  5. Biochemical and molecular characterizaion of Bacillus pumilus isolated from coastal environment in Cochin, India

    Digital Repository Service at National Institute of Oceanography (India)

    Parvathi, A.; Krishna, K.; Jose, J.; Joseph, N.; Nair, S.

    , since the organism is not considered infectious to humans and animals. However, some recent studies have revealed that several Bacillus species including B. pumilus can cause infections, ranging from skin infection to life threatening bacteremia...

  6. Draft Genome Sequence of Bacillus thuringiensis NBIN-866 with High Nematocidal Activity

    OpenAIRE

    Liu, Xiaoyan; Zhou, Ronghua; Fu, Guiping; Zhang, Wei; Min, Yong; Tian, Yuxi; Huang, Daye; Wang, Kaimei; Wan, Zhongyi; Yao, Jingwu; Yang, Ziwen

    2014-01-01

    Bacillus thuringiensis NBIN-866, a Gram-positive bacterium, was isolated from soil in China. We announce here the draft genome sequence of strain B. thuringiensis NBIN-866, which possesses highly nematocidal factors, such as proteins and small molecular peptides.

  7. Genome Sequence of Bacillus anthracis Strain Stendal, Isolated from an Anthrax Outbreak in Cattle in Germany

    OpenAIRE

    Antwerpen, Markus; Elschner, Mandy; Gaede, Wolfgang; Schliephake, Annette; Grass, Gregor; Tomaso, Herbert

    2016-01-01

    In July 2012, an anthrax outbreak occurred among cattle in northern Germany resulting in ten losses. Here, we report the draft genome sequence of Bacillus anthracis strain Stendal, isolated from one of the diseased cows.

  8. Genome Sequence of Bacillus anthracis Strain Stendal, Isolated from an Anthrax Outbreak in Cattle in Germany.

    Science.gov (United States)

    Antwerpen, Markus; Elschner, Mandy; Gaede, Wolfgang; Schliephake, Annette; Grass, Gregor; Tomaso, Herbert

    2016-01-01

    In July 2012, an anthrax outbreak occurred among cattle in northern Germany resulting in ten losses. Here, we report the draft genome sequence ofBacillus anthracisstrain Stendal, isolated from one of the diseased cows. PMID:27056225

  9. Genome Sequence of Bacillus glycinifermentans TH008, Isolated from Ohio Soil

    OpenAIRE

    Zeigler, Daniel R.

    2016-01-01

    The genome sequence of an Ohio soil isolate, TH008, was determined. The sequence reveals a close relationship between TH008 and domesticated Bacillus glycinifermentans strains found in a traditional Korean fermented soybean food.

  10. Invasive Bacillus cereus Infection in a Renal Transplant Patient: A Case Report and Review

    Directory of Open Access Journals (Sweden)

    Susan John

    2012-01-01

    Full Text Available Bacillus cereus is a common cause of gastrointestinal diseases. The majority of individuals with B cereus-related food poisoning recover without any specific treatment. It can, however, rarely cause invasive disease in immunocompromised patients.

  11. Regiospecific Addition of Uracil to Acrylates Catalyzed by Alkaline Protease from Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    Ying CAI; Jian Yi WU; Na WANG; Xiao Feng SUN; Xian Fu LIN

    2004-01-01

    Michael addition reactions of uracil to acrylates were catalyzed by an alkaline protease from Bacillus subtilis in dimethyl sulfoxide at 55 ℃ for 72 h. The adducts were determined by TLC, IR and 1H NMR.

  12. Study of Immunization against Anthrax with the Purified Recombinant Protective Antigen of Bacillus anthracis

    OpenAIRE

    Singh,Yogendra; Ivins, Bruce E.; Leppla, Stephen H.

    1998-01-01

    Protective antigen (PA) of anthrax toxin is the major component of human anthrax vaccine. Currently available human vaccines in the United States and Europe consist of alum-precipitated supernatant material from cultures of toxigenic, nonencapsulated strains of Bacillus anthracis. Immunization with these vaccines requires several boosters and occasionally causes local pain and edema. We previously described the biological activity of a nontoxic mutant of PA expressed in Bacillus subtilis. In ...

  13. Anthrax vaccine design: strategies to achieve comprehensive protection against spore, bacillus, and toxin

    OpenAIRE

    Roehrl, Michael H.; Wang, Jun-Xia

    2005-01-01

    The successful use of Bacillus anthracis as a lethal biological weapon has prompted renewed research interest in the development of more effective vaccines against anthrax. The disease consists of three critical components: spore, bacillus, and toxin, elimination of any of which confers at least partial protection against anthrax. Current remedies rely on postexposure antibiotics to eliminate bacilli and pre- and postexposure vaccination to target primarily toxins. Vaccines effective against ...

  14. Biocontrol and Plant Growth Promotion Characterization of Bacillus Species Isolated from Calendula officinalis Rhizosphere

    OpenAIRE

    Ait Kaki, A.; Kacem Chaouche, N.; Dehimat, L.; Milet, A.; Youcef-Ali, M.; Ongena, Marc; Thonart, Philippe

    2013-01-01

    The phenotypic and genotypic diversity of the plant growth promoting Bacillus genus have been widely investigated in the rhizosphere of various agricultural crops. However, to our knowledge this is the first report on the Bacillus species isolated from the rhizosphere of Calendula officinalis. 15 % of the isolated bacteria were screened for their important antifungal activity against Fusarium oxysporum, Botrytis cinerea, Aspergillus niger, Cladosporium cucumerinium and Alternaria alternata. T...

  15. Genome-wide Screening Reveals the Genetic Determinants of an Antibiotic Insecticide in Bacillus thuringiensis*

    OpenAIRE

    Liu, Xiao-Yan; Ruan, Li-Fang; Hu, Zhen-Fei; Peng, Dong-hai; Cao, Shi-Yun; Yu, Zi-Niu; Liu, Yao; Zheng, Jin-Shui; Sun, Ming

    2010-01-01

    Thuringiensin is a thermostable secondary metabolite in Bacillus thuringiensis and has insecticidal activity against a wide range of insects. Until now, the regulatory mechanisms and genetic determinants involved in thuringiensin production have remained unclear. Here, we successfully used heterologous expression-guided screening in an Escherichia coli–Bacillus thuringiensis shuttle bacterial artificial chromosome library, to clone the intact thuringiensin synthesis (thu) cluster. Then the th...

  16. Evaluation of the Sporicidal Activity of Ethanol Extract of Arctium lappa Root against Bacillus cereus

    OpenAIRE

    Vajihe Karbasizade; Arezoo Dabiri

    2014-01-01

    Background: Bacillus cereus is one of the most common causes of food spoilage, keratitis, endophthalmitis, and panophthalmitis. These bacteria produce spores which are resistant to chemical and physical agents. Nowadays, the sporicidal properties of plants have been considered as alternatives to chemical sporicidal agents. Materials and Methods: In this empirical-experimental study the effect of ethanol extract of edible burdock (Arctium lappa) root has been studied on Bacillus cereus spo...

  17. Biocontrol of Soil Fungi in Tomato with Microencapsulates Containing Bacillus subtilis

    OpenAIRE

    Marcela H. Suarez; Francisco D. Hernandez-Castillo; Gabriel Gallegos-Morales; R. H. Lira-Saldivar; Raul Rodriguez-Herrera; Aguilar, Cristobal N.

    2011-01-01

    Problem statement: An option to reduce pollution by synthetic agro-chemical in root plant disease management is the use of antagonist rhizobacteria belonging to Bacillus genus, because their inhibitory properties, stimulation of plant growth and crop yield increase. Approach: This study was carried out in order to evaluate if Bacillus subtilis strains could play an antagonists role of plant pathogens and if they can be microencapsulated inside a biopolymer matrix. It was adapted an equipment ...

  18. Isolation and Characterization of Coproporphyrin Produced by Four Subspecies of Bacillus thuringiensis

    OpenAIRE

    Harms, R. L.; Martinez, D. R.; Griego, V M

    1986-01-01

    It was found by using spectrophotometric, spectrofluorometric, and high-pressure liquid chromatography that four subspecies of Bacillus thuringiensis produce coproporphyrin. The porphyrin isomer was identified as coproporphyrin I for B. thuringiensis subsp. kurstaki (HD1). The porphyrin was isolated both from spores and from a variety of spent growth media. The quantity of porphyrin released by each Bacillus subspecies differed. The rank order of porphyrin production follows: B. thuringiensis...

  19. Bacillus thuringiensis in Fecal Samples from Greenhouse Workers after Exposure to B. thuringiensis-Based Pesticides

    OpenAIRE

    Jensen, Gert B.; Larsen, Preben; Jacobsen, Bodil L.; Madsen, Bodil; Smidt, Lasse; Andrup, Lars

    2002-01-01

    In a study of occupational exposure to Bacillus thuringiensis, 20 exposed greenhouse workers were examined for Bacillus cereus-like bacteria in fecal samples and on biomonitoring filters. Bacteria with the following characteristics were isolated from eight individuals: intracellular crystalline inclusions characteristic of B. thuringiensis, genes for and production of B. cereus enterotoxins, and positivity for cry11 as determined by PCR. DNA fingerprints of the fecal isolates were identical t...

  20. Complete Genome Sequence of the Endophytic Biocontrol Strain Bacillus velezensis CC09

    Science.gov (United States)

    Cai, Xunchao; Kang, Xingxing; Xi, Huan

    2016-01-01

    Bacillus velezensis is a heterotypic synonym of B. methylotrophicus, B. amyloliquefaciens subsp. plantarum, and Bacillus oryzicola, and has been used to control plant fungal diseases. In order to fully understand the genetic basis of antimicrobial capacities, we did a complete genome sequencing of the endophytic B. velezensis strain CC09. Genes tightly associated with biocontrol ability, including nonribosomal peptide synthetases, polyketide synthetases, iron acquisition, colonization, and volatile organic compound synthesis were identified in the genome. PMID:27688342

  1. Whole-Genome Sequences of 94 Environmental Isolates of Bacillus cereus Sensu Lato

    OpenAIRE

    Van der Auwera, Géraldine A.; Feldgarden, Michael; Kolter, Roberto; Mahillon, Jacques

    2013-01-01

    Bacillus cereus sensu lato is a species complex that includes the anthrax pathogen Bacillus anthracis and other bacterial species of medical, industrial, and ecological importance. Their phenotypes of interest are typically linked to large plasmids that are closely related to the anthrax plasmids pXO1 and pXO2. Here, we present the draft genome sequences of 94 isolates of B. cereus sensu lato, which were chosen for their plasmid content and environmental origins.

  2. Whole-Genome Sequences of 94 Environmental Isolates of Bacillus cereus Sensu Lato

    OpenAIRE

    Van der Auwera, Géraldine A.; Feldgarden, Michael; Kolter, Roberto; Mahillon, Jacques

    2013-01-01

    Bacillus cereus sensu lato is a species complex that includes the anthrax pathogen Bacillus anthracis and other bacterial species of medical, industrial, and ecological importance. Their phenotypes of interest are typically linked to large plasmids that are closely related to the anthrax plasmids pXO1 and pXO2. Here, we present the draft genome sequences of 94 isolates of B. cereus sensu lato, which were chosen for their plasmid content and environmental origins.

  3. Production, purification and characterization of a thermotolerant alkaline serine protease from a novel species Bacillus caseinilyticus

    OpenAIRE

    Mothe, Thirumala; Sultanpuram, Vishnuvardhan Reddy

    2016-01-01

    Alkaline proteases are important enzymes in many industrial applications, especially as additives in laundry detergent industry. Though there are a number of Bacillus species which are reported to be producing proteases, the efficiency of a protease produced by a novel strain has to be studied in comparison to the others. Hence, in this study, an alkaline serine protease produced by a novel species Bacillus caseinilyticus was purified and characterized for its possible usage in detergent indu...

  4. Decontamination Efficacy and Skin Toxicity of Two Decontaminants against Bacillus anthracis

    OpenAIRE

    Chad W Stratilo; Crichton, Melissa K. F.; Sawyer, Thomas W.

    2015-01-01

    Decontamination of bacterial endospores such as Bacillus anthracis has traditionally required the use of harsh or caustic chemicals. The aim of this study was to evaluate the efficacy of a chlorine dioxide decontaminant in killing Bacillus anthracis spores in solution and on a human skin simulant (porcine cadaver skin), compared to that of commonly used sodium hypochlorite or soapy water decontamination procedures. In addition, the relative toxicities of these decontaminants were compared in ...

  5. Molecular characterization of a proteolysis-resistant lipase from Bacillus pumilus SG2

    OpenAIRE

    Sangeetha, R.; Arulpandi, I.; A Geetha

    2014-01-01

    Proteolysis-resistant lipases can be well exploited by industrial processes which employ both lipase and protease as biocatalysts. A proteolysis resistant lipase from Bacillus pumilus SG2 was isolated, purified and characterized earlier. The lipase was resistant to native and commercial proteases. In the present work, we have characterized the lip gene which encodes the proteolysis-resistant lipase from Bacillus pumilus SG2. The parameters and structural details of lipase were analysed. The l...

  6. Modulation of the Bacillus anthracis Secretome by the Immune Inhibitor A1 Protease

    OpenAIRE

    Pflughoeft, Kathryn J.; Swick, Michelle C.; Engler, David A.; Yeo, Hye-Jeong; Koehler, Theresa M.

    2014-01-01

    The Bacillus anthracis secretome includes protective antigen, lethal factor, and edema factor, which are the components of anthrax toxin, and other proteins with known or potential roles in anthrax disease. Immune inhibitor A1 (InhA1) is a secreted metalloprotease that is unique to pathogenic members of the Bacillus genus and has been associated with cleavage of host proteins during infection. Here, we report the effect of InhA1 on the B. anthracis secretome. Differential in-gel electrophores...

  7. Phosphatidylcholine-Specific Phospholipase C and Sphingomyelinase Activities in Bacteria of the Bacillus cereus Group

    OpenAIRE

    Pomerantsev, A. P.; Kalnin, K. V.; Osorio, M.; Leppla, S H

    2003-01-01

    Bacillus anthracis is nonhemolytic, even though it is closely related to the highly hemolytic Bacillus cereus. Hemolysis by B. cereus results largely from the action of phosphatidylcholine-specific phospholipase C (PC-PLC) and sphingomyelinase (SPH), encoded by the plc and sph genes, respectively. In B. cereus, these genes are organized in an operon regulated by the global regulator PlcR. B. anthracis contains a highly similar cereolysin operon, but it is transcriptionally silent because the ...

  8. Complete Genome Sequence of the Endophytic Biocontrol Strain Bacillus velezensis CC09.

    Science.gov (United States)

    Cai, Xunchao; Kang, Xingxing; Xi, Huan; Liu, Changhong; Xue, Yarong

    2016-01-01

    Bacillus velezensis is a heterotypic synonym of B. methylotrophicus, B. amyloliquefaciens subsp. plantarum, and Bacillus oryzicola, and has been used to control plant fungal diseases. In order to fully understand the genetic basis of antimicrobial capacities, we did a complete genome sequencing of the endophytic B. velezensis strain CC09. Genes tightly associated with biocontrol ability, including nonribosomal peptide synthetases, polyketide synthetases, iron acquisition, colonization, and volatile organic compound synthesis were identified in the genome. PMID:27688342

  9. PGPR BACILLUS SPECIES ISOLATED FROM TOMATO PLANT –A COMPARATIVE STUDY ON COCONUT WATER ENRICHMENT

    OpenAIRE

    OS Aysha, P Vinothkumar*, S Vasuki, S Valli, P Nirmala, A Reena

    2012-01-01

    Plant growth promoting rhizobacteria (PGPR) are bacteria that colonize plant roots, they promote plant growth and reduce disease or insect damage. PGPR have been identified within many different bacterial taxa, most commercially developed PGPR for agricultural crops are species of Bacillus which form endospores that confer population stability during formulation and storage of products. Here the rhizobacteria Bacillus sp has been isolated from tomato plant and characterized with routine bioch...

  10. Characterization of AmiBA2446, a Novel Bacteriolytic Enzyme Active against Bacillus Species

    OpenAIRE

    Mehta, Krunal K.; Paskaleva, Elena E.; Azizi-Ghannad, Saba; Ley, Daniel J; Page, Martin A.; Dordick, Jonathan S.; Kane, Ravi S.

    2013-01-01

    There continues to be a need for developing efficient and environmentally friendly treatments for Bacillus anthracis, the causative agent of anthrax. One emerging approach for inactivation of vegetative B. anthracis is the use of bacteriophage endolysins or lytic enzymes encoded by bacterial genomes (autolysins) with highly evolved specificity toward bacterium-specific peptidoglycan cell walls. In this work, we performed in silico analysis of the genome of Bacillus anthracis strain Ames, usin...

  11. A strain-variable bacteriocin in Bacillus anthracis and Bacillus cereus with repeated Cys-Xaa-Xaa motifs

    Directory of Open Access Journals (Sweden)

    Haft Daniel H

    2009-04-01

    Full Text Available Abstract Bacteriocins are peptide antibiotics from ribosomally translated precursors, produced by bacteria often through extensive post-translational modification. Minimal sequence conservation, short gene lengths, and low complexity sequence can hinder bacteriocin identification, even during gene calling, so they are often discovered by proximity to accessory genes encoding maturation, immunity, and export functions. This work reports a new subfamily of putative thiazole-containing heterocyclic bacteriocins. It appears universal in all strains of Bacillus anthracis and B. cereus, but has gone unrecognized because it is always encoded far from its maturation protein operon. Patterns of insertions and deletions among twenty-four variants suggest a repeating functional unit of Cys-Xaa-Xaa. Reviewers This article was reviewed by Andrei Osterman and Lakshminarayan Iyer.

  12. Spore prevalence and toxigenicity of Bacillus cereus and Bacillus thuringiensis isolates from U.S. retail spices.

    Science.gov (United States)

    Hariram, Upasana; Labbé, Ronald

    2015-03-01

    Recent incidents of foodborne illness associated with spices as the vehicle of transmission prompted this examination of U.S. retail spices with regard to Bacillus cereus. This study focused on the levels of aerobic-mesophilic spore-forming bacteria and B cereus spores associated with 247 retail spices purchased from five states in the United States. Samples contained a wide range of aerobic-mesophilic bacterial spore counts (spices had high levels of aerobic spores (> 10(7) CFU/g). Using a novel chromogenic agar, B. cereus and B. thuringiensis spores were isolated from 77 (31%) and 11 (4%) samples, respectively. Levels of B. cereus were spice isolates to form spores, produce diarrheal toxins, and grow at moderately abusive temperatures makes retail spices an important potential vehicle for foodborne illness caused by B. cereus strains, in particular those that produce diarrheal toxins.

  13. Screening of Bacillus thuringiensis strains effective against mosquitoes Prospecção de estirpes de Bacillus thuringiensis efetivas contra mosquitos

    Directory of Open Access Journals (Sweden)

    Rose Gomes Monnerat

    2005-02-01

    Full Text Available The objective of this work was to evaluate 210 Bacillus thuringiensis strains against Aedes aegypti and Culex quinquefasciatus larvae to select the most effective. These strains were isolated from different regions of Brazil and are stored in a Bacillus spp. collection at Embrapa Recursos Genéticos e Biotecnologia, Brasília, Brazil. The selected strains were characterized by morphological (microscopy, biochemical (SDS-PAGE 10% and molecular (PCR methods. Six B. thuringiensis strains were identified as mosquito-toxic after the selective bioassays. None of the strains produced the expected PCR products for detection of cry4, cry11 and cyt1A genes. These results indicate that the activity of mosquitocidal Brazilian strains are not related with Cry4, Cry11 or Cyt proteins, so they could be used as an alternative bioinsecticide against mosquitoes.Neste trabalho foram realizados testes de patogenicidade com 210 estirpes de Bacillus thuringiensis contra larvas de Aedes aegypti e Culex quinquefasciatus, a fim de se determinar as mais eficazes. Estas estirpes foram isoladas de diversas regiões do Brasil e estão armazenadas na coleção de Bacillus spp. da Embrapa Recursos Genéticos e Biotecnologia. As estirpes selecionadas foram caracterizadas por métodos morfológicos (microscopia, bioquímicos (SDS-PAGE 10% e moleculares (Reação em Cadeia da Polimerase. Foram selecionadas seis estirpes entomopatogênicas de Bacillus thuringiensis. Nenhuma das estirpes de Bacillus thuringiensis apresentou produtos de PCR esperados para a detecção dos genes cry4, cry11 e cyt1A. A patogenicidade das estirpes não está associada à presença das toxinas Cry4, Cry11 ou Cyt, assim, essas estirpes poderão ser utilizadas para a formatação de um bioinseticida alternativo contra mosquitos.

  14. Genome characteristics of a novel phage from Bacillus thuringiensis showing high similarity with phage from Bacillus cereus.

    Directory of Open Access Journals (Sweden)

    Yihui Yuan

    Full Text Available Bacillus thuringiensis is an important entomopathogenic bacterium belongs to the Bacillus cereus group, which also includes B. anthracis and B. cereus. Several genomes of phages originating from this group had been sequenced, but no genome of Siphoviridae phage from B. thuringiensis has been reported. We recently sequenced and analyzed the genome of a novel phage, BtCS33, from a B. thuringiensis strain, subsp. kurstaki CS33, and compared the gneome of this phage to other phages of the B. cereus group. BtCS33 was the first Siphoviridae phage among the sequenced B. thuringiensis phages. It produced small, turbid plaques on bacterial plates and had a narrow host range. BtCS33 possessed a linear, double-stranded DNA genome of 41,992 bp with 57 putative open reading frames (ORFs. It had a typical genome structure consisting of three modules: the "late" region, the "lysogeny-lysis" region and the "early" region. BtCS33 exhibited high similarity with several phages, B. cereus phage Wβ and some variants of Wβ, in genome organization and the amino acid sequences of structural proteins. There were two ORFs, ORF22 and ORF35, in the genome of BtCS33 that were also found in the genomes of B. cereus phage Wβ and may be involved in regulating sporulation of the host cell. Based on these observations and analysis of phylogenetic trees, we deduced that B. thuringiensis phage BtCS33 and B. cereus phage Wβ may have a common distant ancestor.

  15. Effects of the electrolytic treatment on Bacillus subtilis Efeito do tratamento eletrolítico em Bacillus subtis

    Directory of Open Access Journals (Sweden)

    Rodolfo Tolentino-Bisneto

    2003-11-01

    Full Text Available Conventional processes of water disinfection can generate toxic composites. It is the case of the trihalomethanes (carcinogenic formed in the contact of chlorine with organic substances present in the water. The electrolytic treatment can be a substitute for the chlorination process without the need for addition of chemical substances to the process. The effect of the electrolytic treatment using carbon cathode on the viability of the microorganism Bacillus subtilis was tested to determine the death process. By means of electronic microscopy, it was observed that the main cause of the microorganism's death was the cellular lysis due to the electroporation in the cell membrane.Processos convencionais de desinfecção de águas podem gerar compostos tóxicos. Esse é o caso dos trialometanos formados na reação do cloro com compostos orgânicos presentes na água. O tratamento eletrolítico pode ser um substituto à cloração com vantagem de não requer a adição de nenhum composto na água. O efeito do tratamento eletrolítico, utilizando eletrodos de carbono, na viabilidade de Bacillus subtilis foi testado para se determinar o mecanismo de morte. Através de microscopia eletrônica, foi possível evidenciar que a morte do microrganismo se deu pela lise celular, provavelmente provocada pela eletroporação irreversível da membrana celular.

  16. Crystal structure of the Bacillus-conserved MazG protein, a nucleotide pyrophosphohydrolase.

    Science.gov (United States)

    Kim, Meong Il; Hong, Minsun

    2016-03-25

    BA1544 from Bacillus anthracis was previously annotated as a transcription factor for the gene cluster ba1554 - ba1558, but has not been experimentally characterized. B. anthracis is an obligate pathogen causing fatal inhalational anthrax, and BA1544 is absolutely conserved in Bacillus species, including Bacillus cereus, Bacillus thuringiensis and Bacillus mycoides, with 100% sequence identity. To address the function of BA1544, we performed structural and biochemical studies, which revealed that BA1544 is a MazG protein. Thus, herein, the protein is defined as Bacillus-conserved MazG (BcMazG). Like other MazG structures, BcMazG assembles into a tetrameric architecture. Each monomer adopts a four-α-helix bundle that accommodates a metal ion using four acidic residues, and presents one putative substrate-binding site. Enzymatic characterization demonstrated that BcMazG is a nucleoside triphosphate (NTP) pyrophosphohydrolase and prefers adenosine triphosphate as a substrate among canonical NTPs. Moreover, structural comparison of BcMazG with its homologues revealed a potential regulation mechanism whereby the enzymatic activity of BcMazG is regulated by its C-terminal region. PMID:26920050

  17. Isolation and Characterization of a New Heterotrophic Nitrifying Bacillus sp. Strain

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Objective To characterize the heterotrophic nitrifying bacteria. Methods The bacteria were isolated from membrane bioreactor for treating synthetic wastewater using the method newly introduced in this study. Fluorescence in situ hybridization (FISH) was used to validate the nonexistence of autotrophic ammonia oxidizers and nitrite oxidizers. Batch tests were carried out to investigate the capability of heterotrophic nitrification by the pure culture. Phylogenetic analysis of the pure culture was performed. Results A heterotrophic nitrifier, named Bacillus sp. LY, was newly isolated from the membrane bioreactor system in which the efficiency of TN removal was up to 80%. After 24-day, incubation, the removal efficiency of COD by Bacillus sp. LYwas 71.7%. The ammonium nitrogen removal rate after assimilation nearly ceased by Bacillus sp. LYwas 74.7%.The phylogenetic tree of Bacillus sp. LY and the neighbouring nitrifiers were given. Conclusions The batch test results indicate that Bacillus sp. LY can utilize the organic carbon as the source of assimilation when it grows on glucose and ammonium chloride medium accompanying the formation of oxidized-nitrogen. It also can denitrify nitrate while nitrifying. Bacillus sp. LY may become a new bacterial resource for heterotrophic nitrification and play a bioremediation role in nutrient removal.

  18. The studies on radiation mutation breeding of Bacillus subtilis with high-yield of amylase

    International Nuclear Information System (INIS)

    The mutagenesis effects on the yield of amylase have been investigated with Bacillus subtilis irradiated by γ-rays and fast neutrons in once or twice irradiation at various dose rates and total irradiation doses. Several parameters such as flat transparent circle, colony diameter, transparent circle diameter and the ratio of flat transparent circle to colony diameter (HC) are used to estimate the radiation mutation of Bacillus subtilis. A series of results has been obtained as (1) Irradiation both with neutrons and γ-rays could make Bacillus subtilis mutationed to produce high-yield amylase effectively. (2) The average colony diameter of Bacillus subtilis irradiated by γ-rays or fast neutrons is smaller than that of control group at various total doses and dose rates. And their colony diameter becomes smaller slightly with the increment of γ-rays irradiation dose. (3) After the second neutrons irradiation, the values of average colony diameter, the biggest colony diameter, average transparent circle diameter and the biggest transparent circle diameter of all mutationed Bacillus subtilis exceed that of original strains greatly. (4) Three kinds of mutationed Bacillus subtilis strains with high-yield amylase have been screened out, in which two strains can produce high-yield amylase steadily after 15 times breeding. Their biggest colony diameter, the biggest transparent circle diameter and the biggest HC value are up to 8.32 mm, 22.38 mm and 5.39 respectively. (authors)

  19. Contribution of Bacillus Isolates to the Flavor Profiles of Vanilla Beans Assessed through Aroma Analysis and Chemometrics

    OpenAIRE

    Fenglin Gu; Yonggan Chen; Yiming Fang; Guiping Wu; Lehe Tan

    2015-01-01

    Colonizing Bacillus in vanilla (Vanilla planifolia Andrews) beans is involved in glucovanillin hydrolysis and vanillin formation during conventional curing. The flavor profiles of vanilla beans under Bacillus-assisted curing were analyzed through gas chromatography-mass spectrometry, electronic nose, and quantitative sensory analysis. The flavor profiles were analytically compared among the vanilla beans under Bacillus-assisted curing, conventional curing, and non-microorganism-assisted curin...

  20. Siderophore-mediated iron acquisition systems in Bacillus cereus: identification of receptors for anthrax virulence-associated petrobactin†a

    OpenAIRE

    Zawadzka, Anna M.; Abergel, Rebecca J.; Nichiporuk, Rita; Andersen, Ulla N.; Raymond, Kenneth N.

    2009-01-01

    During growth under iron limitation, Bacillus cereus and Bacillus anthracis, two human pathogens from the Bacillus cereus group of Gram-positive bacteria, secrete two siderophores, bacillibactin (BB) and petrobactin (PB), for iron acquisition via membrane-associated substrate-binding proteins (SBPs) and other ABC transporter components. Since PB is associated with virulence traits in B. anthracis, the PB-mediated iron uptake system presents a potential target for antimicrobial therapies; its ...

  1. Ecotoxicity studies of antifungal metabolites of Bacillus sp. IBA 33.

    Directory of Open Access Journals (Sweden)

    MA Gordillo

    2015-01-01

    Full Text Available El incremento de enfermedade s de plantas causado por la proliferación de patógenos resis tentes a fungicidas intensificó la investigación de nuevos metabo litos activos contra ellos. El es tudio de métodos biológicos como una a lternativa al control químico h a alcanzado relevancia en años r ecientes. Se ha sugerido que el u so de metabolitos de Bacillus , es una alternativa o un método suplementario a la protección química de las plantas, siempre y cuando ellos no sean tóxicos para los consumidores y sean amiga bles para el medio ambiente. Se ha ev aluado la toxicidad de diferent es concentraciones de metabolito s antifúngicos producidos por Bacillus sp. IBA33. Se investigó su capacidad para inducir efectos citotóxicos mediante diferentes tests como hemólisis e inmovilización de Artemia salina y Allium cepa L. Con 2.56 mg/ml de metabolitos antifúngicos la hemólisis alcanzó 24.07%, para 1 .28 mg/ml fue 14%. La HC 50 fue 10.41 mg/ml. Nauplios de Artemia salina expuestos a 3.2; 1.6 y 0.8 mg/ml de metabolitos antifúngicos mostraron 80; 20 y 10% de morta lidad respectivamente después d e 24 h de tratamiento; la LD 50 fue 2.24 mg/ml. En el test de Allium cepa L después de 72 h de tratamien to, la longitud de las raíces fueron 8.75; 10.35 y 23.75 mm con 3.2; 1.6 y 0.8 mg/ml de metabolitos antifúngicos respectivamente, con una EC 50 de 0.078 mg/ml. Después de 144 h la EC 50 fue 4.11 mg/ml. Solamente con 3.2 mg/ml de metabolitos fue ron observadas aberraciones cromosómicas, vacualización y descentralización de los núcleos en las células de Allium cepa L. No se observaron micronúcleos. Bajo las condiciones experimentales, las concentraciones de metaboli tos antifúngicos ensayadas son consideradas no tóxicas.

  2. Challenges and advances in systems biology analysis of Bacillus spore physiology; molecular differences between an extreme heat resistant spore forming Bacillus subtilis food isolate and a laboratory strain

    NARCIS (Netherlands)

    Brul, S.; Beilen, J.van; Caspers, M.; O'Brien, A.; Koster, C.de; Oomes, S.; Smelt, J.; Kort, R.; Beek, A.ter

    2011-01-01

    Bacterial spore formers are prime organisms of concern in the food industry. Spores from the genus Bacillus are extremely stress resistant, most notably exemplified by high thermotolerance. This sometimes allows surviving spores to germinate and grow out to vegetative cells causing food spoilage and

  3. Spider mite infestations reduce Bacillus thuringiensis toxin concentration in corn leaves and predators avoid spider mites that have fed on Bacillus thuringiensis corn

    Science.gov (United States)

    Transgenic crops containing pyramid-stacked genes for Bacillus thuringiensis derived toxins for controlling coleopteran and lepidopteran pests are increasingly common. As part of environmental risk assessments, these crops are evaluated for toxicity against non-target organisms, and for their poten...

  4. Degradation of proteins during the fermentation of African locust bean (Parkia biglobosa) by strains of Bacillus subtilis and Bacillus pumilus for production of Soumbala

    DEFF Research Database (Denmark)

    Ouoba, L.I.I.; Rechinger, K.B.; Barkholt, Vibeke;

    2003-01-01

    Aims: To examine isolates of Bacillus subtilis and B. pumilus predominant in Soumbala for their ability to degrade African locust bean proteins (ALBP).Methods and Results: Agar diffusion test in casein and ALBP agar was used for screening of isolates. The profiles of water-soluble proteins and fr...

  5. Nutritional optimization for anaerobic growth of Bacillus steaothermophilus LLD-16

    Directory of Open Access Journals (Sweden)

    Muhammad Javed

    2016-04-01

    Full Text Available In this study, a range of nutritional supplements including twenty amino acids, major vitamins and four nucleic acid bases were exploited as added-value supplements for the growth of a lactate-minus (ldh mutant Bacillus stearothermophilus LLD-16 under anaerobic environment. The chemostat studies revealed that five amino acids that includes aspartate, glutamate, isoleucine, methionine, and serine were essential for persuaded growth of B. stearothermophilus LLD-16. The anaerobic batch studies showed that a number of nutritional supplements, such as, p-aminobenzoic acid (PABA, folic acid, pantothenic acid, adenine, glycine, leucine, tryptophan, proline, alanine and α-ketoglutarate, when added individually, improved the biomass levels. In contrast, the higher concentrations of cyanocobalamine or biotin, guanine, uracil and isoleucine were found inhibitory. Furthermore, the study explains why the highest biomass formation cannot necessarily be achieved on the richest mixture of amino acids, and the inadequacy of the biosynthetic machinery is very much dependent on the growth conditions of the microorganism.

  6. Inhibition of Bacillus subtilis growth and sporulation by threonine.

    Science.gov (United States)

    Lamb, D H; Bott, K F

    1979-01-01

    A 1-mg/ml amount of threonine (8.4 mM) inhibited growth and sporulation of Bacillus subtilis 168. Inhibition of sporulation was efficiently reversed by valine and less efficiently by pyruvate, arginine, glutamine, and isoleucine. Inhibition of vegetative growth was reversed by asparate and glutamate as well as by valine, arginine, or glutamine. Cells in minimal growth medium were inhibited only transiently by very high concentrations of threonine, whereas inhibition of sporulation was permanent. Addition of threonine prevented the normal increase in alkaline phosphatase and reduced the production of extracellular protease by about 50%, suggesting that threonine blocked the sporulation process relatively early. 2-Ketobutyrate was able to mimic the effect of threonine on sporulation. Sporulation in a strain selected for resistance to azaleucine was partially resistant. Seventy-five percent of the mutants selected for the ability to grow vegetatively in the presence of high threonine concentrations were found to be simultaneously isoleucine auxotrophs. In at least one of these mutants, the threonine resistance phenotpye could not be dissociated from the isoleucine requirement by transformation. This mutation was closely linked to a known ilvA mutation (recombination index, 0.16). This strain also had reduced intracellular threonine deaminase activity. These results suggest that threonine inhibits B. subtilis by causing valine starvation.

  7. Conversion and degradation of crude oil by Bacillus SP3

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The objective of this study is to demonstrate the basic characteristics of Bacillus SP3 and evaluate its effect on different crude oils. Strain SP3 is a motile, gram-positive, spore-producing rod that was isolated from a reservoir of the Shengli oil field in East China. The cells of strain SP3 grew at high temperatures up to 58℃ at the pH range of 5.5-8.5. Strain SP3 grew facultatively and could use different organic substrates, and produce some metabolites such as 4-hydroxy-4-methyl-2-pentanone, methyl-2-nitrogen benzene and 1, 2-benzenedicarboxylic acid bis ester. Laboratory studies have demonstrated that the strain converted and degraded different components and changed the physical and chemical properties of crude oils. Strain SP3 degraded crude oil and the growth of bacteria on crude oil resulted in loss of aromatic hydrocarbons, resins and asphaltenes. The bioconversion of crude oils would lead to an enrichment in lighter hydrocarbons and an overall redistribution of saturate hydrocarbons. The interactions of microorganisms with crude oils are variable, depending on the microbial species and the chemical compositions of crude oils.

  8. The ecological roles of Bacillus thuringiensis within phyllosphere environments.

    Science.gov (United States)

    Wang, Xiaoxian; Xue, Yarong; Han, Meizhe; Bu, Yuanqing; Liu, Changhong

    2014-08-01

    Bacillus thuringiensis (Bt) is one of the most used bio-control agents to control plant insects, but little is known about its effect on the microbial population and communities on plant leaves. With the culture dependent method, it has been observed that the dynamics of Bt within the phyllosphere varied dependent on both the doses of Bt sprayed on the leaves and the plant species, however, Bt's population size kept stable at about 1000 cfu g(-1) after 15 d since inoculation. By comparing the bacterial abundances and community structures within the phyllosphere of three plant species, we confirmed that Bt at the doses of 1.5×10(7) and 1.5×10(9) cfu mL(-1) respectively did not significantly influence the natural bacterial population size on the leaf surfaces based on culture dependent assay. However, based on culture independent denaturing gradient gel electrophoresis (DGGE), Shannon-Wiener index (H') and Unweighted Pair Group Method with Arithmetic Mean (UPGMA) analysis, Bt has a significant influence on the bacterial communities within the phyllosphere of amaranth and cotton, but not rice. These results indicate that Bt exhibits different behaviors and ecological roles on the microbial diversity within the phyllosphere, and its environmental safety has to be concerned and evaluated in the future. PMID:24534157

  9. Fitness Trade-Offs in Competence Differentiation of Bacillus subtilis

    Science.gov (United States)

    Yüksel, Melih; Power, Jeffrey J.; Ribbe, Jan; Volkmann, Thorsten; Maier, Berenike

    2016-01-01

    In the stationary phase, Bacillus subtilis differentiates stochastically and transiently into the state of competence for transformation (K-state). The latter is associated with growth arrest, and it is unclear how the ability to develop competence is stably maintained, despite its cost. To quantify the effect differentiation has on the competitive fitness of B. subtilis, we characterized the competition dynamics between strains with different probabilities of entering the K-state. The relative fitness decreased with increasing differentiation probability both during the stationary phase and during outgrowth. When exposed to antibiotics inhibiting cell wall synthesis, transcription, and translation, cells that differentiated into the K-state showed a selective advantage compared to differentiation-deficient bacteria; this benefit did not require transformation. Although beneficial, the K-state was not induced by sub-MIC concentrations of antibiotics. Increasing the differentiation probability beyond the wt level did not significantly affect the competition dynamics with transient antibiotic exposure. We conclude that the competition dynamics are very sensitive to the fraction of competent cells under benign conditions but less sensitive during antibiotic exposure, supporting the picture of stochastic differentiation as a fitness trade-off. PMID:27375604

  10. Probing phenotypic growth in expanding Bacillus subtilis biofilms.

    Science.gov (United States)

    Wang, Xiaoling; Koehler, Stephan A; Wilking, James N; Sinha, Naveen N; Cabeen, Matthew T; Srinivasan, Siddarth; Seminara, Agnese; Rubinstein, Shmuel; Sun, Qingping; Brenner, Michael P; Weitz, David A

    2016-05-01

    We develop an optical imaging technique for spatially and temporally tracking biofilm growth and the distribution of the main phenotypes of a Bacillus subtilis strain with a triple-fluorescent reporter for motility, matrix production, and sporulation. We develop a calibration procedure for determining the biofilm thickness from the transmission images, which is based on Beer-Lambert's law and involves cross-sectioning of biofilms. To obtain the phenotype distribution, we assume a linear relationship between the number of cells and their fluorescence and determine the best combination of calibration coefficients that matches the total number of cells for all three phenotypes and with the total number of cells from the transmission images. Based on this analysis, we resolve the composition of the biofilm in terms of motile, matrix-producing, sporulating cells and low-fluorescent materials which includes matrix and cells that are dead or have low fluorescent gene expression. We take advantage of the circular growth to make kymograph plots of all three phenotypes and the dominant phenotype in terms of radial distance and time. To visualize the nonlocal character of biofilm growth, we also make kymographs using the local colonization time. Our technique is suitable for real-time, noninvasive, quantitative studies of the growth and phenotype distribution of biofilms which are either exposed to different conditions such as biocides, nutrient depletion, dehydration, or waste accumulation. PMID:27003268

  11. Glyconanobiotics: Novel carbohydrated nanoparticle antibiotics for MRSA and Bacillus anthracis.

    Science.gov (United States)

    Abeylath, Sampath C; Turos, Edward; Dickey, Sonja; Lim, Daniel V

    2008-03-01

    This report describes the synthesis and evaluation of glycosylated polyacrylate nanoparticles that have covalently-bound antibiotics within their framework. The requisite glycosylated drug monomers were prepared from one of three known antibiotics, an N-sec-butylthio beta-lactam, ciprofloxacin, and a penicillin, by acylation with 3-O-acryloyl-1,2-O-isopropylidene-5,6 bis((chlorosuccinyl)oxy)-d-glucofuranose (7) or 6-O-acetyl-3-O-acryloyl-1,2-O-isopropylidene-5-(chlorosuccinyl)oxy-alpha-d-glucofuranose (10). These acrylated monomers were subjected to emulsion polymerization in a 7:3 (w:w) mixture of butyl acrylate-styrene in the presence of sodium dodecyl sulfate as surfactant (3 weight %) and potassium persulfate as a radical initiator (1 weight %). The resulting nanoparticle emulsions were characterized by dynamic light scattering and found to have similar diameters ( approximately 40 nm) and size distributions to those of our previously studied systems. Microbiological testing showed that the N-sec-butylthio beta-lactam and ciprofloxacin nanoparticles both have powerful in vitro activities against methicillin-resistant Staphylococcus aureus and Bacillus anthracis, while the penicillin-bound nanoparticles have no antimicrobial activity. This indicates the need for matching a suitable antibiotic with the nanoparticle carrier. Overall, the study shows that even relatively large, polar acrylate monomers (MW>1000 amu) can be efficiently incorporated into the nanoparticle matrix by emulsion polymerization, providing opportunities for further advances in nanomedicine. PMID:18063370

  12. Impact of Gastrointestinal Bacillus anthracis Infection on Hepatic B Cells

    Directory of Open Access Journals (Sweden)

    Natacha Colliou

    2015-09-01

    Full Text Available Ingestion of Bacillus anthracis results in rapid gastrointestinal (GI infection, known as GI anthrax. We previously showed that during GI anthrax, there is swift deterioration of intestinal barrier function leading to translocation of gut-associated bacteria into systemic circulation. Additionally, we described dysfunction in colonic B cells. In concordance with our previous studies, here, we report early migration of the Sterne strain of B. anthracis along with other gut-resident bacteria into the infected murine liver. Additionally, despite a global decrease in the B cell population, we observed an increase in both B-1a and marginal zone (MZ-like B cells. Both of these cell types are capable of producing immunoglobulins against common pathogens and commensals, which act as a general antibody barrier before an antigen-specific antibody response. Accumulation of these cells in the liver was associated with an increase in chemokine expression. These data suggest that the presence of Sterne and other commensals in the liver trigger migration of MZ-like B cells from the spleen to the liver to neutralize systemic spread. Further research is required to evaluate the possible cause of their failure to clear the infection within the liver, including the potential role of dysfunctional mitogen-activated protein kinase (MAPK signaling.

  13. Linking Bacillus cereus Genotypes and Carbohydrate Utilization Capacity.

    Science.gov (United States)

    Warda, Alicja K; Siezen, Roland J; Boekhorst, Jos; Wells-Bennik, Marjon H J; de Jong, Anne; Kuipers, Oscar P; Nierop Groot, Masja N; Abee, Tjakko

    2016-01-01

    We characterised carbohydrate utilisation of 20 newly sequenced Bacillus cereus strains isolated from food products and food processing environments and two laboratory strains, B. cereus ATCC 10987 and B. cereus ATCC 14579. Subsequently, genome sequences of these strains were analysed together with 11 additional B. cereus reference genomes to provide an overview of the different types of carbohydrate transporters and utilization systems found in B. cereus strains. The combined application of API tests, defined growth media experiments and comparative genomics enabled us to link the carbohydrate utilisation capacity of 22 B. cereus strains with their genome content and in some cases to the panC phylogenetic grouping. A core set of carbohydrates including glucose, fructose, maltose, trehalose, N-acetyl-glucosamine, and ribose could be used by all strains, whereas utilisation of other carbohydrates like xylose, galactose, and lactose, and typical host-derived carbohydrates such as fucose, mannose, N-acetyl-galactosamine and inositol is limited to a subset of strains. Finally, the roles of selected carbohydrate transporters and utilisation systems in specific niches such as soil, foods and the human host are discussed. PMID:27272929

  14. Screening of Bacillus subtilis transposon mutants with altered riboflavin production.

    Science.gov (United States)

    Tännler, Simon; Zamboni, Nicola; Kiraly, Csilla; Aymerich, Stéphane; Sauer, Uwe

    2008-09-01

    To identify novel targets for metabolic engineering of riboflavin production, we generated about 10,000 random, transposon-tagged mutants of an industrial, riboflavin-producing strain of Bacillus subtilis. Process-relevant screening conditions were established by developing a 96-deep-well plate method with raffinose as the carbon source, which mimics, to some extent, carbon limitation in fed batch cultures. Screening in raffinose and complex LB medium identified more efficiently riboflavin overproducing and underproducing mutants, respectively. As expected for a "loss of function" analysis, most identified mutants were underproducers. Insertion mutants in two genes with yet unknown function, however, were found to attain significantly improved riboflavin titers and yields. These genes and possibly further ones that are related to them are promising candidates for metabolic engineering. While causal links to riboflavin production were not obvious for most underproducers, we demonstrated for the gluconeogenic glyceraldehyde-3-phosphate dehydrogenase GapB how a novel, non-obvious metabolic engineering strategy can be derived from such underproduction mutations. Specifically, we improved riboflavin production on various substrates significantly by deregulating expression of the gluconeogenic genes gapB and pckA through knockout of their genetic repressor CcpN. This improvement was also verified under the more process-relevant conditions of a glucose-limited fed-batch culture. PMID:18582593

  15. The Pore-Forming Haemolysins of Bacillus Cereus: A Review

    Directory of Open Access Journals (Sweden)

    Vincent Sanchis

    2013-06-01

    Full Text Available The Bacillus cereus sensu lato group contains diverse Gram-positive spore-forming bacteria that can cause gastrointestinal diseases and severe eye infections in humans. They have also been incriminated in a multitude of other severe, and frequently fatal, clinical infections, such as osteomyelitis, septicaemia, pneumonia, liver abscess and meningitis, particularly in immuno-compromised patients and preterm neonates. The pathogenic properties of this organism are mediated by the synergistic effects of a number of virulence products that promote intestinal cell destruction and/or resistance to the host immune system. This review focuses on the pore-forming haemolysins produced by B. cereus: haemolysin I (cereolysin O, haemolysin II, haemolysin III and haemolysin IV (CytK. Haemolysin I belongs to the cholesterol-dependent cytolysin (CDC family whose best known members are listeriolysin O and perfringolysin O, produced by L. monocytogenes and C. perfringens respectively. HlyII and CytK are oligomeric ß-barrel pore-forming toxins related to the α-toxin of S. aureus or the ß-toxin of C. perfringens. The structure of haemolysin III, the least characterized haemolytic toxin from the B. cereus, group has not yet been determined.

  16. Chemodiversity of cereulide, the emetic toxin of Bacillus cereus.

    Science.gov (United States)

    Marxen, Sandra; Stark, Timo D; Frenzel, Elrike; Rütschle, Andrea; Lücking, Genia; Pürstinger, Gabriel; Pohl, Elena E; Scherer, Siegfried; Ehling-Schulz, Monika; Hofmann, Thomas

    2015-03-01

    Food-borne intoxications are increasingly caused by the dodecadepsipeptide cereulide, the emetic toxin produced by Bacillus cereus. As such intoxications pose a health risk to humans, a more detailed understanding on the chemodiversity of this toxin is mandatory for the reliable risk assessment of B. cereus toxins in foods. Mass spectrometric screening now shows a series of at least 18 cereulide variants, among which the previously unknown isocereulides A-G were determined for the first time by means of UPLC-TOF MS and ion-trap MS(n) sequencing, (13)C-labeling experiments, and post-hydrolytic dipeptide and enantioselective amino acid analysis. The data demonstrate a high microheterogeneity in cereulide and show evidence for a relaxed proof reading function of the non-ribosomal cereulide peptide synthetase complex giving rise to an enhanced cereulide chemodiversity. Most intriguingly, the isocereulides were found to differ widely in their cell toxicity correlating with their ionophoric properties (e.g., purified isocereulide A showed about 8-fold higher cytotoxicity than purified cereulide in the HEp-2 assay and induced an immediate breakdown of bilayer membranes). These findings provide a substantial contribution to the knowledge-based risk assessment of B. cereus toxins in foods, representing a still unsolved challenge in the field of food intoxications.

  17. Bacillus thuringiensis membrane-damaging toxins acting on mammalian cells.

    Science.gov (United States)

    Celandroni, Francesco; Salvetti, Sara; Senesi, Sonia; Ghelardi, Emilia

    2014-12-01

    Bacillus thuringiensis is widely used as a biopesticide in forestry and agriculture, being able to produce potent species-specific insecticidal toxins and considered nonpathogenic to other animals. More recently, however, repeated observations are documenting the association of this microorganism with various infectious diseases in humans, such as food-poisoning-associated diarrheas, periodontitis, bacteremia, as well as ocular, burn, and wound infections. Similar to B. cereus, B. thuringiensis produces an array of virulence factors acting against mammalian cells, such as phosphatidylcholine- and phosphatidylinositol-specific phospholipase C (PC-PLC and PI-PLC), hemolysins, in particular hemolysin BL (HBL), and various enterotoxins. The contribution of some of these toxins to B. thuringiensis pathogenicity has been studied in animal models of infection, following intravitreous, intranasal, or intratracheal inoculation. These studies lead to the speculation that the activities of PC-PLC, PI-PLC, and HBL are responsible for most of the pathogenic properties of B. thuringiensis in nongastrointestinal infections in mammals. This review summarizes data regarding the biological activity, the genetic basis, and the structural features of these membrane-damaging toxins.

  18. Bacillus thuringiensis: fermentation process and risk assessment: a short review

    Directory of Open Access Journals (Sweden)

    Deise M. F Capalbo

    1995-02-01

    Full Text Available Several factors make the local production of Bacillus thuringiensis (Bt highly appropriate for pest control in developing nations. Bt can be cheaply produced on a wide variety of low cost, organic substrates. Local production results in considerable savings in hard currency which otherwise would be spent on importation of chemical and biological insecticides. The use of Bt in Brazil has been limited in comparison with chemical insecticides. Although Bt is imported, some Brazilian researchers have been working on its development and production. Fermentation processes (submerged and semi-solid were applied, using by-products from agro-industries. As the semi-solid fermentation process demonstrated to be interesting for Bt endotoxins production, it could be adopted for small scale local production. Although promising results had been achieved, national products have not been registered due to the absence of a specific legislation for biological products. Effective actions are being developed in order to solve this gap. Regardless of the biocontrol agents being considered atoxic and harmless to the environment, information related to direct and indirect effects of microbials are still insufficient in many cases. The risk analysis of the use of microbial control agents is of upmost importance nowadays, and is also discussed.

  19. A biochemically active MCM-like helicase in Bacillus cereus

    Science.gov (United States)

    Samuels, Martin; Gulati, Gaurav; Shin, Jae-Ho; Opara, Rejoice; McSweeney, Elizabeth; Sekedat, Matt; Long, Stephen; Kelman, Zvi; Jeruzalmi, David

    2009-01-01

    The mini-chromosome maintenance (MCM) proteins serve as the replicative helicases in archaea and eukaryotes. Interestingly, an MCM homolog was identified, by BLAST analysis, within a phage integrated in the bacterium Bacillus cereus (Bc). BcMCM is only related to the AAA region of MCM-helicases; the typical amino-terminus is missing and is replaced by a segment with weak homology to primases. We show that BcMCM displays 3′→5′ helicase and ssDNA-stimulated ATPase activity, properties that arise from its conserved AAA domain. Isolated BcMCM is a monomer in solution but likely forms the functional oligomer in vivo. We found that the BcMCM amino-terminus can bind ssDNA and harbors a zinc atom, both hallmarks of the typical MCM amino-terminus. No BcMCM-catalyzed primase activity could be detected. We propose that the divergent amino-terminus of BcMCM is a paralog of the corresponding region of MCM-helicases. A divergent amino terminus makes BcMCM a useful model for typical MCM-helicases since it accomplishes the same function using an apparently unrelated structure. PMID:19474351

  20. Monarch larvae sensitivity to Bacillus thuringiensis- purified proteins and pollen.

    Science.gov (United States)

    Hellmich, R L; Siegfried, B D; Sears, M K; Stanley-Horn, D E; Daniels, M J; Mattila, H R; Spencer, T; Bidne, K G; Lewis, L C

    2001-10-01

    Laboratory tests were conducted to establish the relative toxicity of Bacillus thuringiensis (Bt) toxins and pollen from Bt corn to monarch larvae. Toxins tested included Cry1Ab, Cry1Ac, Cry9C, and Cry1F. Three methods were used: (i) purified toxins incorporated into artificial diet, (ii) pollen collected from Bt corn hybrids applied directly to milkweed leaf discs, and (iii) Bt pollen contaminated with corn tassel material applied directly to milkweed leaf discs. Bioassays of purified Bt toxins indicate that Cry9C and Cry1F proteins are relatively nontoxic to monarch first instars, whereas first instars are sensitive to Cry1Ab and Cry1Ac proteins. Older instars were 12 to 23 times less susceptible to Cry1Ab toxin compared with first instars. Pollen bioassays suggest that pollen contaminants, an artifact of pollen processing, can dramatically influence larval survival and weight gains and produce spurious results. The only transgenic corn pollen that consistently affected monarch larvae was from Cry1Ab event 176 hybrids, currently corn planted and for which re-registration has not been applied. Results from the other types of Bt corn suggest that pollen from the Cry1Ab (events Bt11 and Mon810) and Cry1F, and experimental Cry9C hybrids, will have no acute effects on monarch butterfly larvae in field settings. PMID:11559841

  1. Process development and intensification for enhanced production of Bacillus lipopeptides.

    Science.gov (United States)

    Rangarajan, Vivek; Clarke, Kim G

    2015-01-01

    The growing interest in Bacillus lipopeptides for high-value applications has driven process design, development and optimization for enhanced lipopeptide production. Traditional optimization approaches have been directed towards improving the overall titres by modification of media components and environmental parameters, almost exclusively in submerged cultures. Carbon and nitrogen sources, trace elements and oxygen availability have all been demonstrated to exhibit significant influences on lipopeptide yield, productivity and selectivity. This insight into process-linked kinetics, especially selectivity, has led to the introduction of novel process intensification and integration strategies which further promote process efficiency, and which include foam fractionation, inverse fluidization, rotating disc contacting and microfiltration with recycle. These strategies have not only transformed the production capabilities, but have also successfully integrated upstream production with downstream purification through cell retention and in situ product removal. This review analyses and critically discusses the impact of process conditions and process optimization strategies for improving lipopeptide production kinetics, specifically highlighting the emerging trend of process intensification and integration strategies and further, proposes a heuristic route to enhance lipopeptide production.

  2. Microbial forensics: fiber optic microarray subtyping of Bacillus anthracis

    Science.gov (United States)

    Shepard, Jason R. E.

    2009-05-01

    The past decade has seen increased development and subsequent adoption of rapid molecular techniques involving DNA analysis for detection of pathogenic microorganisms, also termed microbial forensics. The continued accumulation of microbial sequence information in genomic databases now better positions the field of high-throughput DNA analysis to proceed in a more manageable fashion. The potential to build off of these databases exists as technology continues to develop, which will enable more rapid, cost effective analyses. This wealth of genetic information, along with new technologies, has the potential to better address some of the current problems and solve the key issues involved in DNA analysis of pathogenic microorganisms. To this end, a high density fiber optic microarray has been employed, housing numerous DNA sequences simultaneously for detection of various pathogenic microorganisms, including Bacillus anthracis, among others. Each organism is analyzed with multiple sequences and can be sub-typed against other closely related organisms. For public health labs, real-time PCR methods have been developed as an initial preliminary screen, but culture and growth are still considered the gold standard. Technologies employing higher throughput than these standard methods are better suited to capitalize on the limitless potential garnered from the sequence information. Microarray analyses are one such format positioned to exploit this potential, and our array platform is reusable, allowing repetitive tests on a single array, providing an increase in throughput and decrease in cost, along with a certainty of detection, down to the individual strain level.

  3. Nutrient depletion in Bacillus subtilis biofilms triggers matrix production

    International Nuclear Information System (INIS)

    Many types of bacteria form colonies that grow into physically robust and strongly adhesive aggregates known as biofilms. A distinguishing characteristic of bacterial biofilms is an extracellular polymeric substance (EPS) matrix that encases the cells and provides physical integrity to the colony. The EPS matrix consists of a large amount of polysaccharide, as well as protein filaments, DNA and degraded cellular materials. The genetic pathways that control the transformation of a colony into a biofilm have been widely studied, and yield a spatiotemporal heterogeneity in EPS production. Spatial gradients in metabolites parallel this heterogeneity in EPS, but nutrient concentration as an underlying physiological initiator of EPS production has not been explored. Here, we study the role of nutrient depletion in EPS production in Bacillus subtilis biofilms. By monitoring simultaneously biofilm size and matrix production, we find that EPS production increases at a critical colony thickness that depends on the initial amount of carbon sources in the medium. Through studies of individual cells in liquid culture we find that EPS production can be triggered at the single-cell level by reducing nutrient concentration. To connect the single-cell assays with conditions in the biofilm, we calculate carbon concentration with a model for the reaction and diffusion of nutrients in the biofilm. This model predicts the relationship between the initial concentration of carbon and the thickness of the colony at the point of internal nutrient deprivation. (paper)

  4. Photoprotection of Bacillus thuringiensis kurstaki from ultraviolet irradiation

    International Nuclear Information System (INIS)

    Irradiation of Bacillus thuringiensis var. kurstaki HD1 at 300-350 nm for up to 12 hr using a photochemical reactor results in a rapid loss of its toxicity to larvae of Heliothis armigera. Photoprotection of the toxic component was obtained by adsorption of cationic chromophores such as acriflavin (AF), methyl green, and rhodamine B to B. thuringiensis. AF gave the best photoprotection and a level of 0.42 mmol/g dye absorbed per gram of B. thuringiensis was highly toxic even after 12 hr of ultraviolet (uv) irradiation as compared to the control (77.5 and 5% of insect mortality, respectively). Ultraviolet and Fourier-transform infrared spectroscopic studies indicate molecular interactions between B. thuringiensis and AF. The nature of these interactions and energy or charge transfer as possible mechanisms of photoprotection are discussed. It is speculated that tryptophan residues are essential for the toxic effect of B. thuringiensis. It is suggested that photoprotection is attained as energy is transferred from the excited tryptophan moieties to the chromophore molecules

  5. Setting risk-informed environmental standards for Bacillus anthracis spores.

    Science.gov (United States)

    Hong, Tao; Gurian, Patrick L; Ward, Nicholas F Dudley

    2010-10-01

    In many cases, human health risk from biological agents is associated with aerosol exposures. Because air concentrations decline rapidly after a release, it may be necessary to use concentrations found in other environmental media to infer future or past aerosol exposures. This article presents an approach for linking environmental concentrations of Bacillus. anthracis (B. anthracis) spores on walls, floors, ventilation system filters, and in human nasal passages with human health risk from exposure to B. anthracis spores. This approach is then used to calculate example values of risk-informed concentration standards for both retrospective risk mitigation (e.g., prophylactic antibiotics) and prospective risk mitigation (e.g., environmental clean up and reoccupancy). A large number of assumptions are required to calculate these values, and the resulting values have large uncertainties associated with them. The values calculated here suggest that documenting compliance with risks in the range of 10(-4) to 10(-6) would be challenging for small diameter (respirable) spore particles. For less stringent risk targets and for releases of larger diameter particles (which are less respirable and hence less hazardous), environmental sampling would be more promising.

  6. Metabolic engineering of Bacillus subtilis for terpenoid production.

    Science.gov (United States)

    Guan, Zheng; Xue, Dan; Abdallah, Ingy I; Dijkshoorn, Linda; Setroikromo, Rita; Lv, Guiyuan; Quax, Wim J

    2015-11-01

    Terpenoids are the largest group of small-molecule natural products, with more than 60,000 compounds made from isopentenyl diphosphate (IPP) and its isomer dimethylallyl diphosphate (DMAPP). As the most diverse group of small-molecule natural products, terpenoids play an important role in the pharmaceutical, food, and cosmetic industries. For decades, Escherichia coli (E. coli) and Saccharomyces cerevisiae (S. cerevisiae) were extensively studied to biosynthesize terpenoids, because they are both fully amenable to genetic modifications and have vast molecular resources. On the other hand, our literature survey (20 years) revealed that terpenoids are naturally more widespread in Bacillales. In the mid-1990s, an inherent methylerythritol phosphate (MEP) pathway was discovered in Bacillus subtilis (B. subtilis). Since B. subtilis is a generally recognized as safe (GRAS) organism and has long been used for the industrial production of proteins, attempts to biosynthesize terpenoids in this bacterium have aroused much interest in the scientific community. This review discusses metabolic engineering of B. subtilis for terpenoid production, and encountered challenges will be discussed. We will summarize some major advances and outline future directions for exploiting the potential of B. subtilis as a desired "cell factory" to produce terpenoids.

  7. Bacillus anthracis genome organization in light of whole transcriptome sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Martin, Jeffrey; Zhu, Wenhan; Passalacqua, Karla D.; Bergman, Nicholas; Borodovsky, Mark

    2010-03-22

    Emerging knowledge of whole prokaryotic transcriptomes could validate a number of theoretical concepts introduced in the early days of genomics. What are the rules connecting gene expression levels with sequence determinants such as quantitative scores of promoters and terminators? Are translation efficiency measures, e.g. codon adaptation index and RBS score related to gene expression? We used the whole transcriptome shotgun sequencing of a bacterial pathogen Bacillus anthracis to assess correlation of gene expression level with promoter, terminator and RBS scores, codon adaptation index, as well as with a new measure of gene translational efficiency, average translation speed. We compared computational predictions of operon topologies with the transcript borders inferred from RNA-Seq reads. Transcriptome mapping may also improve existing gene annotation. Upon assessment of accuracy of current annotation of protein-coding genes in the B. anthracis genome we have shown that the transcriptome data indicate existence of more than a hundred genes missing in the annotation though predicted by an ab initio gene finder. Interestingly, we observed that many pseudogenes possess not only a sequence with detectable coding potential but also promoters that maintain transcriptional activity.

  8. Mutagenesis of Bacillus subtilis spores exposed to simulated space environment

    Science.gov (United States)

    Munakata, N.; Natsume, T.; Takahashi, K.; Hieda, K.; Panitz, C.; Horneck, G.

    Bacterial spores can endure in a variety of extreme earthly environments. However, some conditions encountered during the space flight could be detrimental to DNA in the spore, delimiting the possibility of transpermia. We investigate the genetic consequences of the exposure to space environments in a series of preflight simulation project of EXPOSE. Using Bacillus subtilis spores of repair-proficient HA101 and repair-deficient TKJ6312 strains, the mutations conferring resistance to rifampicin were detected, isolated and sequenced. Most of the mutations were located in a N-terminal region of the rpoB gene encoding RNA polymerase beta-subunit. Among several potentially mutagenic factors, high vacuum, UV radiation, heat, and accelerated heavy ions induced mutations with varying efficiencies. A majority of mutations induced by vacuum exposure carried a tandem double-base change (CA to TT) at a unique sequence context of TCAGC. Results indicate that the vacuum and high temperature may act synergistically for the induction of mutations.

  9. Linking Bacillus cereus Genotypes and Carbohydrate Utilization Capacity

    Science.gov (United States)

    Warda, Alicja K.; Siezen, Roland J.; Boekhorst, Jos; Wells-Bennik, Marjon H. J.; de Jong, Anne; Kuipers, Oscar P.; Nierop Groot, Masja N.; Abee, Tjakko

    2016-01-01

    We characterised carbohydrate utilisation of 20 newly sequenced Bacillus cereus strains isolated from food products and food processing environments and two laboratory strains, B. cereus ATCC 10987 and B. cereus ATCC 14579. Subsequently, genome sequences of these strains were analysed together with 11 additional B. cereus reference genomes to provide an overview of the different types of carbohydrate transporters and utilization systems found in B. cereus strains. The combined application of API tests, defined growth media experiments and comparative genomics enabled us to link the carbohydrate utilisation capacity of 22 B. cereus strains with their genome content and in some cases to the panC phylogenetic grouping. A core set of carbohydrates including glucose, fructose, maltose, trehalose, N-acetyl-glucosamine, and ribose could be used by all strains, whereas utilisation of other carbohydrates like xylose, galactose, and lactose, and typical host-derived carbohydrates such as fucose, mannose, N-acetyl-galactosamine and inositol is limited to a subset of strains. Finally, the roles of selected carbohydrate transporters and utilisation systems in specific niches such as soil, foods and the human host are discussed. PMID:27272929

  10. Inactivation of Bacillus Subtilis by Atomic Oxygen Radical Anion

    Institute of Scientific and Technical Information of China (English)

    LI Longchun; WANG Lian; YU Zhou; LV Xuanzhong; LI Quanxin

    2007-01-01

    UAtomic oxygen radical anion (O- ) is one of the most active oxygen species, and has extremely high oxidation ability toward small-molecules of hydrocarbons. However, to our knowledge, little is known about the effects of O- on cells of micro-organisms. This work showed that O- could quickly react with the Bacillus subtilis cells and seriously damage the cell walls a s well as their other contents, leading to a fast and irreversible inactivation. SEM micrographs revealed that the cell structures were dramatically destroyed by their exposure to O-. The inactivation efficiencies of B. subtilis depend on the O-- intensity, the initial population of cells and the treatment temperature, but not on the pH in the range of our investigation. For a cell concentration of 106 cfu/ml, the number of survived cells dropped from 106 cfu/ml to 103 cfu/ml after about five-minute irradiation by an O- flux in an intensity of 233 nA/cm2 under a dry argon environment (30 ℃, 1 atm, exposed size: 1.8 cm2). The inactivation mechanism of micro-organisms induced by O- is also discussed.

  11. Dynamics of Aerial Tower Formation in Bacillus subtilis Biofilms

    Science.gov (United States)

    Sinha, Naveen; Seminara, Agnese; Wilking, James; Brenner, Michael; Weitz, Dave

    2012-02-01

    Biofilms are highly-organized colonies of bacteria that form on surfaces. These colonies form sophisticated structures which make them robust and difficult to remove from environments such as catheters, where they pose serious infection problems. Previous work has shown that sub-mm sized aerial towers form on the surface of Bacillus subtilis colony biofilms. Spore-formation is located preferentially at the tops of these towers, known as fruiting bodies, which aid in the dispersal and propagation of the colony to new sites. The formation of towers is strongly affected by the quorum-sensing molecule surfactin and the cannibalism pathway of the bacteria. In the present work, we use confocal fluorescence microscopy to study the development of individual fruiting bodies, allowing us to visualize the time-dependent spatial distribution of matrix-forming and sporulating bacteria within the towers. With this information, we investigate the physical mechanisms, such as surface tension and polymer concentration gradients, that drive the formation of these structures.

  12. Biocalcifying Bacillus subtilis cells effectively consolidate deteriorated Globigerina limestone.

    Science.gov (United States)

    Micallef, Roderick; Vella, Daniel; Sinagra, Emmanuel; Zammit, Gabrielle

    2016-07-01

    Microbially induced calcite precipitation occurs naturally on ancient limestone surfaces in Maltese hypogea. We exploited this phenomenon and treated deteriorated limestone with biocalcifying bacteria. The limestone was subjected to various mechanical and physical tests to present a statistically robust data set to prove that treatment was indeed effective. Bacillus subtilis conferred uniform bioconsolidation to a depth of 30 mm. Drilling resistance values were similar to those obtained for freshly quarried limestone (9 N) and increased up to 15 N. Treatment resulted in a high resistance to salt deterioration and a slow rate of water absorption. The overall percentage porosity of treated limestone varied by ±6 %, thus the pore network was preserved. We report an eco-friendly treatment that closely resembles the mineral composition of limestone and that penetrates into the porous structure without affecting the limestones' natural properties. The treatment is of industrial relevance since it compares well with stone consolidants available commercially. PMID:27072564

  13. Molecular characterization of the circulating Bacillus anthracis in Jordan.

    Science.gov (United States)

    Aqel, Amin Abdelfattah; Hailat, Ekhlas; Serrecchia, Luigina; Aqel, Suad; Campese, Emanuele; Vicari, Nadia; Fasanella, Antonio

    2015-12-01

    To understand the biomolecular charcteristics of Bacillus anthracis in Jordan, 20 blood smear slides from dead animals with suspected anthrax were analyzed using conventional and molecular approaches. All slides were positive for B. anthracis by conventional staining but no growth of the organism on selective media was detected. However, of the 20 samples, 16 were B. anthracis DNA-positive using polymerase chain reaction (PCR). Seven samples provided enough quantity and quality of DNA, and their multilocus variable tandem repeat analysis (MLVA)-15 loci analysis revealed two different genotypes. All genotypes were belonging to A.B..r. 008/009 which is very common in Asia and Europe. Single nucleotide repeat (SNR) analysis revealed that there were no sub genotypes. Molecular diagnosis of animal anthrax in Jordan is not used routinely; henceforth, official diagnosis of anthrax is based on the observation of the slides by optical microscope and this can often cause reading errors. Therefore, the prevalence of the disease in Jordan might be slightly lower than that reported by the official bodies. PMID:26156620

  14. Crystal structure of Bacillus anthracis transpeptidase enzyme CapD.

    Energy Technology Data Exchange (ETDEWEB)

    Wu, R.; Richter, S.; Zhang, R.; Anderson, V. J.; Missiakas, D.; Joachimiak, A.; Biosciences Division; Univ. of Chicago

    2009-09-04

    Bacillus anthracis elaborates a poly-{gamma}-d-glutamic acid capsule that protects bacilli from phagocytic killing during infection. The enzyme CapD generates amide bonds with peptidoglycan cross-bridges to anchor capsular material within the cell wall envelope of B. anthracis. The capsular biosynthetic pathway is essential for virulence during anthrax infections and can be targeted for anti-infective inhibition with small molecules. Here, we present the crystal structures of the {gamma}-glutamyltranspeptidase CapD with and without {alpha}-l-Glu-l-Glu dipeptide, a non-hydrolyzable analog of poly-{gamma}-d-glutamic acid, in the active site. Purified CapD displays transpeptidation activity in vitro, and its structure reveals an active site broadly accessible for poly-{gamma}-glutamate binding and processing. Using structural and biochemical information, we derive a mechanistic model for CapD catalysis whereby Pro{sup 427}, Gly{sup 428}, and Gly{sup 429} activate the catalytic residue of the enzyme, Thr{sup 352}, and stabilize an oxyanion hole via main chain amide hydrogen bonds.

  15. Production of Bioactive Compounds by Bacillus subtilis against Sclerotium rolfsii

    Directory of Open Access Journals (Sweden)

    Nalisha, I.

    2006-01-01

    Full Text Available This study aims to investigate the characteristic of bioactive compound produced by Bacillus subtilis against Sclerotium rolfsii and the influence of additive supplements on the antagonistic activity of B. subtilis. The fact that B. subtilis produced an antifungal substance which has inhibitory effect on wide range of fungi, including S. rolfsii, is well known. To learn the effect of pH, temperature and light condition on the production of antifungal compound, B. subtilis was inoculated in Potato Dextrose Broth at various initial pH, temperatures and light conditions, respectively. This antagonist was found to produce antifungal compound that stable at 80C with 58.3 % inhibition on S. rolfsii. The activity was constant within a wide range of pH (3–11. However, treatment with pH11 lead to higher antifungal activity (31.57 % inhibition and it was also found to produce substance that can endure dark condition (46.24 % inhibition with fungicidal effect on S. rolfsii. A series of experiments also been carried out to enhance the antifungal production by supplementing different carbon source preparation into bacterial liquid culture. B. subtilis were grown in minimal medium containing 1 % of oil palm root, Ganoderma lucidum or chitin, respectively prior to bioassay. Crude culture from oil palm root supplemented culture shown significantly reduction in S. rolfsii growth compared to other carbon source crude culture or the antagonism alone, suggesting that this approach may provide improved biocontrol efficiency.

  16. Loop grafting of Bacillus subtilis lipase A: inversion of enantioselectivity.

    Science.gov (United States)

    Boersma, Ykelien L; Pijning, Tjaard; Bosma, Margriet S; van der Sloot, Almer M; Godinho, Luís F; Dröge, Melloney J; Winter, Remko T; van Pouderoyen, Gertie; Dijkstra, Bauke W; Quax, Wim J

    2008-08-25

    Lipases are successfully applied in enantioselective biocatalysis. Most lipases contain a lid domain controlling access to the active site, but Bacillus subtilis Lipase A (LipA) is a notable exception: its active site is solvent exposed. To improve the enantioselectivity of LipA in the kinetic resolution of 1,2-O-isopropylidene-sn-glycerol (IPG) esters, we replaced a loop near the active-site entrance by longer loops originating from Fusarium solani cutinase and Penicillium purpurogenum acetylxylan esterase, thereby aiming to increase the interaction surface for the substrate. The resulting loop hybrids showed enantioselectivities inverted toward the desired enantiomer of IPG. The acetylxylan esterase-derived variant showed an inversion in enantiomeric excess (ee) from -12.9% to +6.0%, whereas the cutinase-derived variant was improved to an ee of +26.5%. The enantioselectivity of the cutinase-derived variant was further improved by directed evolution to an ee of +57.4%. PMID:18721749

  17. Genetic analysis of petrobactin transport in Bacillus anthracis.

    Science.gov (United States)

    Carlson, Paul E; Dixon, Shandee D; Janes, Brian K; Carr, Katherine A; Nusca, Tyler D; Anderson, Erica C; Keene, Sarra E; Sherman, David H; Hanna, Philip C

    2010-02-01

    Iron acquisition mechanisms play an important role in the pathogenesis of many infectious microbes. In Bacillus anthracis, the siderophore petrobactin is required for both growth in iron-depleted conditions and for full virulence of the bacterium. Here we demonstrate the roles of two putative petrobactin binding proteins FatB and FpuA (encoded by GBAA5330 and GBAA4766 respectively) in B. anthracis iron acquisition and pathogenesis. Markerless deletion mutants were created using allelic exchange. The Delta fatB strain was capable of wild-type levels of growth in iron-depleted conditions, indicating that FatB does not play an essential role in petrobactin uptake. In contrast, Delta fpuA bacteria exhibited a significant decrease in growth under low-iron conditions when compared with wild-type bacteria. This mutant could not be rescued by the addition of exogenous purified petrobactin. Further examination of this strain demonstrated increased levels of petrobactin accumulation in the culture supernatants, suggesting no defect in siderophore synthesis or export but, instead, an inability of Delta fpuA to import this siderophore. Delta fpuA spores were also significantly attenuated in a murine model of inhalational anthrax. These results provide the first genetic evidence demonstrating the role of FpuA in petrobactin uptake. PMID:20487286

  18. Thermo Stability Study of Crude Amylase from Bacillus Isolate

    Directory of Open Access Journals (Sweden)

    Moti Lal

    2015-09-01

    Full Text Available An amylolytic strain was collected from rotten potato and its activity evaluated. The isolated strain was cultivated for amylase production in shake flasks at 35±2oC and the fermentation pattern was studied. Optimum temperature for maximum enzyme synthesis was observed at 35°C, when initial pH of fermentation medium was adjusted to 5.5. Maximum extracellular amylase activity of 7.9 U/mL and the maximum intracellular activity of 320 U/mL was recorded. Although maximum biomass was present at 12.6 g/L but highest growth rate was observed between 08 to 40h with maximum at 36h. The extracellular amylase present in the broth was partially purified with an overall yield of 44% through purification procedure of ammonium sulphate precipitation. After completed extraction and partial purification and stabilization, the stability of enzyme was observed in a range of temperature and pH between 60°C-90°C and 2-8 pH respectively. Maximum enzyme activity was demonstrated at 90°C, and pH of 5.5 and 6.5. The thermo stability of the amylases of this Bacillus species was comparable to that of amylases from other microbial sources.

  19. [Asymmetric biosynthesis of d-pseudoephedrine by recombinant Bacillus subtilis].

    Science.gov (United States)

    Peng, Yanhong; Zhang, Liang; Ding, Zhongyang; Wang, Zhengxiang; Shi, Guiyang

    2011-07-01

    In order to successfully express the carbonyl reductase gene mldh in Bacillus subtilis and complete coenzyme regeneration by B. subtilis glucose dehydrogenase, the promoter PrpsD and the terminator TrpsD from B. subtilis rpsD gene were used as the expression cassette to be a recombinant plasmid pHY300plk-PrpsD-TrpsD. After that, the carbonyl reductase gene mldh was inserted into the previous plasmid and a plasmid pHY300plk-PrpsD-mldh-TrpsD was achieved, followed by transformed into B. subtilis Wb600 to obtain a recombinant B. subtilis Wb600 (pHY300plk-PrpsD-mldh-TrpsD). Subsequently, the results for whole-cell biotransformation from recombinant B. subtilis showed that it could be used to catalyze MAK (1-phenyl- 1-keto-2-methylaminopropane) to d-pseudoephedrine in the presence of glucose. The yield of d-pseudoephedrine could be up to 97.5 mg/L and the conversion rate of MAK was 24.1%. This study indicates the possibility of biotransformation production of d-pseudoephedrine from recombinant B. subtilis.

  20. Quantum dot incorporated Bacillus spore as nanosensor for viral infection.

    Science.gov (United States)

    Zhang, Xinya; Zhou, Qian; Shen, Zhongfeng; Li, Zheng; Fei, Ruihua; Ji, Eoon Hye; Hu, Shen; Hu, Yonggang

    2015-12-15

    In this paper, we report a high-throughput biological method to prepare spore-based monodisperse microparticles (SMMs) and then form the nanocomposites of CdTe quantum dot (QD)-loaded SMMs by utilizing the endogenous functional groups from Bacillus spores. The SMMs and QD-incorporated spore microspheres (QDSMs) were characterized by using transmission electron microscopy, high-resolution transmission electron microscopy, fluorescence microscopy, fluorescence and UV-visible absorption spectroscopy, zeta potential analysis, Fourier-transform infrared spectroscopy, potentiometric titrations, X-ray photo-electron spectroscopy. The thermodynamics of QD/SMM interaction and antigen/QDSM interaction was also investigated by isothermal titration microcalorimetry (ITC). Fluorescent QDSMs coded either with a single luminescence color or with multiple colors of controlled emission intensity ratios were obtained. Green QDSMs were used as a model system to detect porcine parvovirus antibody in swine sera via flow cytometry, and the results demonstrated a great potential of QDSMs in high-throughput immunoassays. Due to the advantages such as simplicity, low cost, high throughput and eco-friendliness, our developed platform may find wide applications in disease detection, food safety evaluation and environmental assessment.

  1. Comparison of hand hygiene procedures for removing Bacillus cereus spores.

    Science.gov (United States)

    Sasahara, Teppei; Hayashi, Shunji; Hosoda, Kouichi; Morisawa, Yuji; Hirai, Yoshikazu

    2014-01-01

    Bacillus cereus is a spore-forming bacterium. B. cereus occasionally causes nosocomial infections, in which hand contamination with the spores plays an important role. Therefore, hand hygiene is the most important practice for controlling nosocomial B. cereus infections. This study aimed to determine the appropriate hand hygiene procedure for removing B. cereus spores. Thirty volunteers' hands were experimentally contaminated with B. cereus spores, after which they performed 6 different hand hygiene procedures. We compared the efficacy of the procedures in removing the spores from hands. The alcohol-based hand-rubbing procedures scarcely removed them. The soap washing procedures reduced the number of spores by more than 2 log10. Extending the washing time increased the spore-removing efficacy of the washing procedures. There was no significant difference in efficacy between the use of plain soap and antiseptic soap. Handwashing with soap is appropriate for removing B. cereus spores from hands. Alcohol-based hand-rubbing is not effective. PMID:25252644

  2. Continuous evolution of Bacillus thuringiensis toxins overcomes insect resistance.

    Science.gov (United States)

    Badran, Ahmed H; Guzov, Victor M; Huai, Qing; Kemp, Melissa M; Vishwanath, Prashanth; Kain, Wendy; Nance, Autumn M; Evdokimov, Artem; Moshiri, Farhad; Turner, Keith H; Wang, Ping; Malvar, Thomas; Liu, David R

    2016-05-01

    The Bacillus thuringiensis δ-endotoxins (Bt toxins) are widely used insecticidal proteins in engineered crops that provide agricultural, economic, and environmental benefits. The development of insect resistance to Bt toxins endangers their long-term effectiveness. Here we have developed a phage-assisted continuous evolution selection that rapidly evolves high-affinity protein-protein interactions, and applied this system to evolve variants of the Bt toxin Cry1Ac that bind a cadherin-like receptor from the insect pest Trichoplusia ni (TnCAD) that is not natively bound by wild-type Cry1Ac. The resulting evolved Cry1Ac variants bind TnCAD with high affinity (dissociation constant Kd = 11-41 nM), kill TnCAD-expressing insect cells that are not susceptible to wild-type Cry1Ac, and kill Cry1Ac-resistant T. ni insects up to 335-fold more potently than wild-type Cry1Ac. Our findings establish that the evolution of Bt toxins with novel insect cell receptor affinity can overcome insect Bt toxin resistance and confer lethality approaching that of the wild-type Bt toxin against non-resistant insects. PMID:27120167

  3. Role of receptors in Bacillus thuringiensis crystal toxin activity.

    Science.gov (United States)

    Pigott, Craig R; Ellar, David J

    2007-06-01

    Bacillus thuringiensis produces crystalline protein inclusions with insecticidal or nematocidal properties. These crystal (Cry) proteins determine a particular strain's toxicity profile. Transgenic crops expressing one or more recombinant Cry toxins have become agriculturally important. Individual Cry toxins are usually toxic to only a few species within an order, and receptors on midgut epithelial cells have been shown to be critical determinants of Cry specificity. The best characterized of these receptors have been identified for lepidopterans, and two major receptor classes have emerged: the aminopeptidase N (APN) receptors and the cadherin-like receptors. Currently, 38 different APNs have been reported for 12 different lepidopterans. Each APN belongs to one of five groups that have unique structural features and Cry-binding properties. While 17 different APNs have been reported to bind to Cry toxins, only 2 have been shown to mediate toxin susceptibly in vivo. In contrast, several cadherin-like proteins bind to Cry toxins and confer toxin susceptibility in vitro, and disruption of the cadherin gene has been associated with toxin resistance. Nonetheless, only a small subset of the lepidopteran-specific Cry toxins has been shown to interact with cadherin-like proteins. This review analyzes the interactions between Cry toxins and their receptors, focusing on the identification and validation of receptors, the molecular basis for receptor recognition, the role of the receptor in resistant insects, and proposed models to explain the sequence of events at the cell surface by which receptor binding leads to cell death. PMID:17554045

  4. Fitness Trade-Offs in Competence Differentiation of Bacillus subtilis.

    Science.gov (United States)

    Yüksel, Melih; Power, Jeffrey J; Ribbe, Jan; Volkmann, Thorsten; Maier, Berenike

    2016-01-01

    In the stationary phase, Bacillus subtilis differentiates stochastically and transiently into the state of competence for transformation (K-state). The latter is associated with growth arrest, and it is unclear how the ability to develop competence is stably maintained, despite its cost. To quantify the effect differentiation has on the competitive fitness of B. subtilis, we characterized the competition dynamics between strains with different probabilities of entering the K-state. The relative fitness decreased with increasing differentiation probability both during the stationary phase and during outgrowth. When exposed to antibiotics inhibiting cell wall synthesis, transcription, and translation, cells that differentiated into the K-state showed a selective advantage compared to differentiation-deficient bacteria; this benefit did not require transformation. Although beneficial, the K-state was not induced by sub-MIC concentrations of antibiotics. Increasing the differentiation probability beyond the wt level did not significantly affect the competition dynamics with transient antibiotic exposure. We conclude that the competition dynamics are very sensitive to the fraction of competent cells under benign conditions but less sensitive during antibiotic exposure, supporting the picture of stochastic differentiation as a fitness trade-off. PMID:27375604

  5. Continuous evolution of Bacillus thuringiensis toxins overcomes insect resistance.

    Science.gov (United States)

    Badran, Ahmed H; Guzov, Victor M; Huai, Qing; Kemp, Melissa M; Vishwanath, Prashanth; Kain, Wendy; Nance, Autumn M; Evdokimov, Artem; Moshiri, Farhad; Turner, Keith H; Wang, Ping; Malvar, Thomas; Liu, David R

    2016-05-01

    The Bacillus thuringiensis δ-endotoxins (Bt toxins) are widely used insecticidal proteins in engineered crops that provide agricultural, economic, and environmental benefits. The development of insect resistance to Bt toxins endangers their long-term effectiveness. Here we have developed a phage-assisted continuous evolution selection that rapidly evolves high-affinity protein-protein interactions, and applied this system to evolve variants of the Bt toxin Cry1Ac that bind a cadherin-like receptor from the insect pest Trichoplusia ni (TnCAD) that is not natively bound by wild-type Cry1Ac. The resulting evolved Cry1Ac variants bind TnCAD with high affinity (dissociation constant Kd = 11-41 nM), kill TnCAD-expressing insect cells that are not susceptible to wild-type Cry1Ac, and kill Cry1Ac-resistant T. ni insects up to 335-fold more potently than wild-type Cry1Ac. Our findings establish that the evolution of Bt toxins with novel insect cell receptor affinity can overcome insect Bt toxin resistance and confer lethality approaching that of the wild-type Bt toxin against non-resistant insects.

  6. Bacillus thuringiensis Toxins: An Overview of Their Biocidal Activity

    Science.gov (United States)

    Palma, Leopoldo; Muñoz, Delia; Berry, Colin; Murillo, Jesús; Caballero, Primitivo

    2014-01-01

    Bacillus thuringiensis (Bt) is a Gram positive, spore-forming bacterium that synthesizes parasporal crystalline inclusions containing Cry and Cyt proteins, some of which are toxic against a wide range of insect orders, nematodes and human-cancer cells. These toxins have been successfully used as bioinsecticides against caterpillars, beetles, and flies, including mosquitoes and blackflies. Bt also synthesizes insecticidal proteins during the vegetative growth phase, which are subsequently secreted into the growth medium. These proteins are commonly known as vegetative insecticidal proteins (Vips) and hold insecticidal activity against lepidopteran, coleopteran and some homopteran pests. A less well characterized secretory protein with no amino acid similarity to Vip proteins has shown insecticidal activity against coleopteran pests and is termed Sip (secreted insecticidal protein). Bin-like and ETX_MTX2-family proteins (Pfam PF03318), which share amino acid similarities with mosquitocidal binary (Bin) and Mtx2 toxins, respectively, from Lysinibacillus sphaericus, are also produced by some Bt strains. In addition, vast numbers of Bt isolates naturally present in the soil and the phylloplane also synthesize crystal proteins whose biological activity is still unknown. In this review, we provide an updated overview of the known active Bt toxins to date and discuss their activities. PMID:25514092

  7. Raman spectroscopy of Bacillus thuringiensis physiology and inactivation

    Science.gov (United States)

    Morrow, J. B.; Almeida, J.; Cole, K. D.; Reipa, V.

    2012-12-01

    The ability to detect spore contamination and inactivation is relevant to developing and determining decontamination strategy success for food and water safety. This study was conducted to develop a systematic comparison of nondestructive vibrational spectroscopy techniques (Surface-Enhanced Raman Spectroscopy, SERS, and normal Raman) to determine indicators of Bacillus thuringiensis physiology (spore, vegetative, outgrown, germinated and inactivated spore forms). SERS was found to provide better resolution of commonly utilized signatures of spore physiology (dipicolinic acid at 1006 cm-1 and 1387 cm-1) compared to normal Raman and native fluorescence indigenous to vegetative and outgrown cell samples was quenched in SERS experiment. New features including carotenoid pigments (Raman features at 1142 cm-1, 1512 cm-1) were identified for spore cell forms. Pronounced changes in the low frequency region (300 cm-1 to 500 cm-1) in spore spectra occurred upon germination and inactivation (with both free chlorine and by autoclaving) which is relevant to guiding decontamination and detection strategies using Raman techniques.

  8. Adhesion of Spores of Bacillus thuringiensis on a Planar Surface

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Eunhyea [Georgia Institute of Technology; Kweon, Hyojin [Georgia Institute of Technology; Yiacoumi, Sotira [Georgia Institute of Technology; Lee, Ida [University of Tennessee, Knoxville (UTK); Joy, David Charles [ORNL; Palumbo, Anthony Vito [ORNL; Tsouris, Costas [ORNL

    2010-01-01

    Adhesion of spores of Bacillus thuringiensis (Bt) and spherical silica particles on surfaces was experimentally and theoretically investigated in this study. Topography analysis via atomic force microscopy (AFM) and electron microscopy indicates that Bt spores are rod shaped, {approx}1.3 {mu}m in length and {approx}0.8 {mu}m in diameter. The adhesion force of Bt spores and silica particles on gold-coated glass was measured at various relative humidity (RH) levels by AFM. It was expected that the adhesion force would vary with RH because the individual force components contributing to the adhesion force depend on RH. The adhesion force between a particle and a planar surface in atmospheric environments was modeled as the contribution of three major force components: capillary, van der Waals, and electrostatic interaction forces. Adhesion force measurements for Bt spore (silica particle) and the gold surface system were comparable with calculations. Modeling results show that there is a critical RH value, which depends on the hydrophobicity of the materials involved, below which the water meniscus does not form and the contribution of the capillary force is zero. As RH increases, the van der Waals force decreases while the capillary force increases to a maximum value.

  9. Photoprotection of Bacillus thuringiensis kurstaki from ultraviolet irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Cohen, E.; Rozen, H.; Joseph, T.; Braun, S.; Margulies, L. (Department of Entomology, Faculty of Agriculture, Hebrew University of Jerusalem, Rehovot (Israel))

    1991-05-01

    Irradiation of Bacillus thuringiensis var. kurstaki HD1 at 300-350 nm for up to 12 hr using a photochemical reactor results in a rapid loss of its toxicity to larvae of Heliothis armigera. Photoprotection of the toxic component was obtained by adsorption of cationic chromophores such as acriflavin (AF), methyl green, and rhodamine B to B. thuringiensis. AF gave the best photoprotection and a level of 0.42 mmol/g dye absorbed per gram of B. thuringiensis was highly toxic even after 12 hr of ultraviolet (uv) irradiation as compared to the control (77.5 and 5% of insect mortality, respectively). Ultraviolet and Fourier-transform infrared spectroscopic studies indicate molecular interactions between B. thuringiensis and AF. The nature of these interactions and energy or charge transfer as possible mechanisms of photoprotection are discussed. It is speculated that tryptophan residues are essential for the toxic effect of B. thuringiensis. It is suggested that photoprotection is attained as energy is transferred from the excited tryptophan moieties to the chromophore molecules.

  10. Midgut microbiota and host immunocompetence underlie Bacillus thuringiensis killing mechanism.

    Science.gov (United States)

    Caccia, Silvia; Di Lelio, Ilaria; La Storia, Antonietta; Marinelli, Adriana; Varricchio, Paola; Franzetti, Eleonora; Banyuls, Núria; Tettamanti, Gianluca; Casartelli, Morena; Giordana, Barbara; Ferré, Juan; Gigliotti, Silvia; Ercolini, Danilo; Pennacchio, Francesco

    2016-08-23

    Bacillus thuringiensis is a widely used bacterial entomopathogen producing insecticidal toxins, some of which are expressed in insect-resistant transgenic crops. Surprisingly, the killing mechanism of B. thuringiensis remains controversial. In particular, the importance of the septicemia induced by the host midgut microbiota is still debated as a result of the lack of experimental evidence obtained without drastic manipulation of the midgut and its content. Here this key issue is addressed by RNAi-mediated silencing of an immune gene in a lepidopteran host Spodoptera littoralis, leaving the midgut microbiota unaltered. The resulting cellular immunosuppression was characterized by a reduced nodulation response, which was associated with a significant enhancement of host larvae mortality triggered by B. thuringiensis and a Cry toxin. This was determined by an uncontrolled proliferation of midgut bacteria, after entering the body cavity through toxin-induced epithelial lesions. Consequently, the hemolymphatic microbiota dramatically changed upon treatment with Cry1Ca toxin, showing a remarkable predominance of Serratia and Clostridium species, which switched from asymptomatic gut symbionts to hemocoelic pathogens. These experimental results demonstrate the important contribution of host enteric flora in B. thuringiensis-killing activity and provide a sound foundation for developing new insect control strategies aimed at enhancing the impact of biocontrol agents by reducing the immunocompetence of the host.

  11. SinR controls enterotoxin expression in Bacillus thuringiensis biofilms.

    Directory of Open Access Journals (Sweden)

    Annette Fagerlund

    Full Text Available The entomopathogen Bacillus thuringiensis produces dense biofilms under various conditions. Here, we report that the transition phase regulators Spo0A, AbrB and SinR control biofilm formation and swimming motility in B. thuringiensis, just as they control biofilm formation and swarming motility in the closely related saprophyte species B. subtilis. However, microarray analysis indicated that in B. thuringiensis, in contrast to B. subtilis, SinR does not control an eps operon involved in exopolysaccharides production, but regulates genes involved in the biosynthesis of the lipopeptide kurstakin. This lipopeptide is required for biofilm formation and was previously shown to be important for survival in the host cadaver (necrotrophism. Microarray analysis also revealed that the SinR regulon contains genes coding for the Hbl enterotoxin. Transcriptional fusion assays, Western blots and hemolysis assays confirmed that SinR controls Hbl expression, together with PlcR, the main virulence regulator in B. thuringiensis. We show that Hbl is expressed in a sustained way in a small subpopulation of the biofilm, whereas almost all the planktonic population transiently expresses Hbl. The gene coding for SinI, an antagonist of SinR, is expressed in the same biofilm subpopulation as hbl, suggesting that hbl transcription heterogeneity is SinI-dependent. B. thuringiensis and B. cereus are enteric bacteria which possibly form biofilms lining the host intestinal epithelium. Toxins produced in biofilms could therefore be delivered directly to the target tissue.

  12. PROFILIN ACTIVATES BACILLUS THURINGIENSIS PHOSPHOINOSITIDE SPECIFIC PHOSPHOLIPASE C

    Directory of Open Access Journals (Sweden)

    Sandeepta Burgula

    2012-08-01

    Full Text Available Many extracellular signaling molecules including hormones, growth factors, neurotransmitters andimmunoglobulins elicit intracellular responses by activating phosphatidylinositol-specific phospholipase C (PI-PLCupon binding to their cell surface receptors. Activated PLC catalyses the hydrolysis of Phosphotidylinositol 4,5-bisphosphate (PIP2 to generate DAG and IP3 , which act as signaling molecules that control various cellular processes.Exploring the mechanism of regulation of PLC activity may lead to understanding various signaling events thatregulate cell growth and differentiation. One of the dramatic effects of profilin is inhibition of PIP2 hydrolysis by PLC-γ in eukaryotic cells. In the present study, the effect of profilin on Phosphotidylinositol specific phospholipase C (PI-PLC purified from Bacillus thuringiensis (Bt was examined. Assay of PI-PLC activity indicated that Bovine profilinactivated the hydrolysis of phosphotidylinositol (PI by BtPI-PLC in a concentration dependent manner under in vitroconditions. A 250 % increase in activity was noted in the presence of profilin but not in presence of phosphoprofilin. Inthe presence of profilin more proteins are observed in the soluble fraction. In conclusion it can be stated that thatprofilin activates bacterial PLC activity towards PI hydrolysis

  13. Degradation of ochratoxin A by Bacillus amyloliquefaciens ASAG1.

    Science.gov (United States)

    Chang, Xiaojiao; Wu, Zidan; Wu, Songling; Dai, Yanshi; Sun, Changpo

    2015-01-01

    Ochratoxin A (OTA) is widely found in food and feed products as a mycotoxin contaminant. It is produced by Penicillium species and several Aspergillus species. The identification OTA detoxification microorganisms is believed to be the best approach for decontamination. In this study, we isolated ASAG1, a bacterium with the ability to degrade OTA effectively, from grain depot-stored maize. A 16S rDNA sequencing approach was used to identify this strain as Bacillus amyloliquefaciens ASAG1. The degradation of OTA was detected in both medium and cell-free extracts after incubation with a culture of B. amyloliquefaciens ASAG1 cells. Subsequently, a hydrolysed enzyme (carboxypeptidase) related to the enzymatic conversion of OTA was cloned from the B. amyloliquefaciens ASAG1 genome. Using the Escherichia coli Expression System, we successfully expressed and purified this carboxypeptidase. When this enzyme was incubated with the engineered recombinant E. coli cells, the concentration of OTA was dramatically degraded. Our data therefore indicate that the carboxypeptidase produced by B. amyloliquefaciens ASAG1 is likely responsible for the biodegradation of OTA.

  14. A novel antifungal protein of Bacillus subtilis B25.

    Science.gov (United States)

    Tan, Zhiqiong; Lin, Baoying; Zhang, Rongyi

    2013-01-01

    Bacillus subtilis B25 was isolated from banana rhizosphere soil. It has been confirmed for B25 to have stronger antagonism against Fusarium oxysporum f.sp.cubense, Additionally B25 has good inhibitory to plant pathogens, including Corynespora cassiicola, Alternaria solani, Botrytis cinerea and Colletotrichum gloeosporioides on potato dextrose agar (PDA) plates. The antagonistic substance can be extracted from cell-free culture broth supernatants by 70% (w/v) (NH4)2 SO4 saturation. Clear blank band was observed between the protein and a pathogen. The examination of antagonistic mechanism under light microscope showed that the antifungal protein of B25 appeared to inhibit pathogens by leading to mycelium and spores tumescence, distortion, abnormality. The isolation procedure comprised ion exchange chromatography on DEAE-Sephadex Fast Flow and gel filtration chromatography on SephadexG-100. The purified antifungal fraction showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The active fraction was identified by NanoLC-ESI-MS/MS The amino acid sequences of 17 peptides segments were obtained. The analysis of the protein suggested that it was a hypothetical protein (gi154685475), with a relative molecular mass of 38708.67 Da and isoelectric point (pI) of 5.63. PMID:24255843

  15. Linking Bacillus cereus Genotypes and Carbohydrate Utilization Capacity.

    Directory of Open Access Journals (Sweden)

    Alicja K Warda

    Full Text Available We characterised carbohydrate utilisation of 20 newly sequenced Bacillus cereus strains isolated from food products and food processing environments and two laboratory strains, B. cereus ATCC 10987 and B. cereus ATCC 14579. Subsequently, genome sequences of these strains were analysed together with 11 additional B. cereus reference genomes to provide an overview of the different types of carbohydrate transporters and utilization systems found in B. cereus strains. The combined application of API tests, defined growth media experiments and comparative genomics enabled us to link the carbohydrate utilisation capacity of 22 B. cereus strains with their genome content and in some cases to the panC phylogenetic grouping. A core set of carbohydrates including glucose, fructose, maltose, trehalose, N-acetyl-glucosamine, and ribose could be used by all strains, whereas utilisation of other carbohydrates like xylose, galactose, and lactose, and typical host-derived carbohydrates such as fucose, mannose, N-acetyl-galactosamine and inositol is limited to a subset of strains. Finally, the roles of selected carbohydrate transporters and utilisation systems in specific niches such as soil, foods and the human host are discussed.

  16. Bacillus pumilus Cyanide Dihydratase Mutants with Higher Catalytic Activity.

    Science.gov (United States)

    Crum, Mary A; Sewell, B Trevor; Benedik, Michael J

    2016-01-01

    Cyanide degrading nitrilases are noted for their potential to detoxify industrial wastewater contaminated with cyanide. However, such application would benefit from an improvement to characteristics such as their catalytic activity and stability. Following error-prone PCR for random mutagenesis, several cyanide dihydratase mutants from Bacillus pumilus were isolated based on improved catalysis. Four point mutations, K93R, D172N, A202T, and E327K were characterized and their effects on kinetics, thermostability and pH tolerance were studied. K93R and D172N increased the enzyme's thermostability whereas E327K mutation had a less pronounced effect on stability. The D172N mutation also increased the affinity of the enzyme for its substrate at pH 7.7 but lowered its k cat. However, the A202T mutation, located in the dimerization or the A surface, destabilized the protein and abolished its activity. No significant effect on activity at alkaline pH was observed for any of the purified mutants. These mutations help confirm the model of CynD and are discussed in the context of the protein-protein interfaces leading to the protein quaternary structure. PMID:27570524

  17. Scalable purification of Bacillus anthracis protective antigen from Escherichia coli.

    Science.gov (United States)

    Gwinn, William; Zhang, Mei; Mon, Sandii; Sampey, Darryl; Zukauskas, David; Kassebaum, Corby; Zmuda, Jonathan F; Tsai, Amos; Laird, Michael W

    2006-01-01

    The anthrax toxin consists of three proteins, protective antigen (PA), lethal factor, and edema factor that are produced by the Gram-positive bacterium, Bacillus anthracis. Current vaccines against anthrax use PA as their primary component. In this study, we developed a scalable process to produce and purify multi-gram quantities of highly pure, recombinant PA (rPA) from Escherichia coli. The rPA protein was produced in a 50-L fermentor and purified to >99% purity using anion-exchange, hydrophobic interaction, and hydroxyapatite chromatography. The final yield of purified rPA from medium cell density fermentations resulted in approximately 2.7 g of rPA per kg of cell paste (approximately 270 mg/L) of highly pure, biologically active rPA protein. The results presented here exhibit the ability to generate multi-gram quantities of rPA from E. coli that may be used for the development of new anthrax vaccines and anthrax therapeutics. PMID:15935696

  18. A part toolbox to tune genetic expression in Bacillus subtilis.

    Science.gov (United States)

    Guiziou, Sarah; Sauveplane, Vincent; Chang, Hung-Ju; Clerté, Caroline; Declerck, Nathalie; Jules, Matthieu; Bonnet, Jerome

    2016-09-01

    Libraries of well-characterised components regulating gene expression levels are essential to many synthetic biology applications. While widely available for the Gram-negative model bacterium Escherichia coli, such libraries are lacking for the Gram-positive model Bacillus subtilis, a key organism for basic research and biotechnological applications. Here, we engineered a genetic toolbox comprising libraries of promoters, Ribosome Binding Sites (RBS), and protein degradation tags to precisely tune gene expression in B. subtilis We first designed a modular Expression Operating Unit (EOU) facilitating parts assembly and modifications and providing a standard genetic context for gene circuits implementation. We then selected native, constitutive promoters of B. subtilis and efficient RBS sequences from which we engineered three promoters and three RBS sequence libraries exhibiting ∼14 000-fold dynamic range in gene expression levels. We also designed a collection of SsrA proteolysis tags of variable strength. Finally, by using fluorescence fluctuation methods coupled with two-photon microscopy, we quantified the absolute concentration of GFP in a subset of strains from the library. Our complete promoters and RBS sequences library comprising over 135 constructs enables tuning of GFP concentration over five orders of magnitude, from 0.05 to 700 μM. This toolbox of regulatory components will support many research and engineering applications in B. subtilis. PMID:27402159

  19. Midgut microbiota and host immunocompetence underlie Bacillus thuringiensis killing mechanism.

    Science.gov (United States)

    Caccia, Silvia; Di Lelio, Ilaria; La Storia, Antonietta; Marinelli, Adriana; Varricchio, Paola; Franzetti, Eleonora; Banyuls, Núria; Tettamanti, Gianluca; Casartelli, Morena; Giordana, Barbara; Ferré, Juan; Gigliotti, Silvia; Ercolini, Danilo; Pennacchio, Francesco

    2016-08-23

    Bacillus thuringiensis is a widely used bacterial entomopathogen producing insecticidal toxins, some of which are expressed in insect-resistant transgenic crops. Surprisingly, the killing mechanism of B. thuringiensis remains controversial. In particular, the importance of the septicemia induced by the host midgut microbiota is still debated as a result of the lack of experimental evidence obtained without drastic manipulation of the midgut and its content. Here this key issue is addressed by RNAi-mediated silencing of an immune gene in a lepidopteran host Spodoptera littoralis, leaving the midgut microbiota unaltered. The resulting cellular immunosuppression was characterized by a reduced nodulation response, which was associated with a significant enhancement of host larvae mortality triggered by B. thuringiensis and a Cry toxin. This was determined by an uncontrolled proliferation of midgut bacteria, after entering the body cavity through toxin-induced epithelial lesions. Consequently, the hemolymphatic microbiota dramatically changed upon treatment with Cry1Ca toxin, showing a remarkable predominance of Serratia and Clostridium species, which switched from asymptomatic gut symbionts to hemocoelic pathogens. These experimental results demonstrate the important contribution of host enteric flora in B. thuringiensis-killing activity and provide a sound foundation for developing new insect control strategies aimed at enhancing the impact of biocontrol agents by reducing the immunocompetence of the host. PMID:27506800

  20. The impact of secondary pests on Bacillus thuringiensis (Bt) crops.

    Science.gov (United States)

    Catarino, Rui; Ceddia, Graziano; Areal, Francisco J; Park, Julian

    2015-06-01

    The intensification of agriculture and the development of synthetic insecticides enabled worldwide grain production to more than double in the last third of the 20th century. However, the heavy dependence and, in some cases, overuse of insecticides has been responsible for negative environmental and ecological impacts across the globe, such as a reduction in biodiversity, insect resistance to insecticides, negative effects on nontarget species (e.g. natural enemies) and the development of secondary pests. The use of recombinant DNA technology to develop genetically engineered insect-resistant crops could mitigate many of the negative side effects of insecticides. One such genetic alteration enables crops to express toxic crystalline (Cry) proteins from the soil bacteria Bacillus thuringiensis (Bt). Despite the widespread adoption of Bt crops, there are still a range of unanswered questions concerning longer term agro-ecosystem interactions. For instance, insect species that are not susceptible to the expressed toxin can develop into secondary pests and cause significant damage to the crop. Here, we review the main causes surrounding secondary pest dynamics in Bt crops and the impact of such outbreaks. Regardless of the causes, if nonsusceptible secondary pest populations exceed economic thresholds, insecticide spraying could become the immediate solution at farmers' disposal, and the sustainable use of this genetic modification technology may be in jeopardy. Based on the literature, recommendations for future research are outlined that will help to improve the knowledge of the possible long-term ecological trophic interactions of employing this technology. PMID:25832330

  1. Fitness trade-offs in competence differentiation of Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Melih Yüksel

    2016-06-01

    Full Text Available In the stationary phase, Bacillus subtilis differentiates stochastically and transiently into the state of competence for transformation (K-state. The latter is associated with growth arrest, and it is unclear how the ability to develop competence is stably maintained, despite its cost. To quantify the effect differentiation has on the competitive fitness of B. subtilis, we characterized the competition dynamics between strains with different probabilities of entering the K-state. The relative fitness decreased with increasing differentiation probability both during the stationary phase and during outgrowth. When exposed to antibiotics inhibiting cell wall synthesis, transcription, and translation, cells that differentiated into the K-state showed a selective advantage compared to differentiation-deficient bacteria; this benefit did not require transformation. Although beneficial, the K-state was not induced by sub-MIC concentrations of antibiotics. Increasing the differentiation probability beyond the wt level did not significantly affect the competition dynamics with transient antibiotic exposure. We conclude that the competition dynamics are very sensitive to the fraction of competent cells under benign conditions but less sensitive during antibiotic exposure, supporting the picture of stochastic differentiation as a fitness trade-off.

  2. Degradation of ochratoxin A by Bacillus amyloliquefaciens ASAG1.

    Science.gov (United States)

    Chang, Xiaojiao; Wu, Zidan; Wu, Songling; Dai, Yanshi; Sun, Changpo

    2015-01-01

    Ochratoxin A (OTA) is widely found in food and feed products as a mycotoxin contaminant. It is produced by Penicillium species and several Aspergillus species. The identification OTA detoxification microorganisms is believed to be the best approach for decontamination. In this study, we isolated ASAG1, a bacterium with the ability to degrade OTA effectively, from grain depot-stored maize. A 16S rDNA sequencing approach was used to identify this strain as Bacillus amyloliquefaciens ASAG1. The degradation of OTA was detected in both medium and cell-free extracts after incubation with a culture of B. amyloliquefaciens ASAG1 cells. Subsequently, a hydrolysed enzyme (carboxypeptidase) related to the enzymatic conversion of OTA was cloned from the B. amyloliquefaciens ASAG1 genome. Using the Escherichia coli Expression System, we successfully expressed and purified this carboxypeptidase. When this enzyme was incubated with the engineered recombinant E. coli cells, the concentration of OTA was dramatically degraded. Our data therefore indicate that the carboxypeptidase produced by B. amyloliquefaciens ASAG1 is likely responsible for the biodegradation of OTA. PMID:25517039

  3. Vacuum Distillation Residue Upgrading by an Indigenous Bacillus Cereus

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    Mitra Sadat Tabatabaee

    2013-07-01

    Full Text Available Background:Biological processing of heavy fractions of crude oils offers less severe process conditions and higher selectivity for refining. Biochemical Processes are expected to be low demand energy processes and certainly ecofriendly.Results:A strain of biosurfactant producing bacterium was isolated from an oil contaminated soil at Tehran refinery distillation unit. Based on selected phenotypic and genotypic characteristic including morphology, biochemical proprety, and 16 SrRNA sequencing identified as a novel strain of Bacillus cereus (JQ178332. This bacterium endures a wide range of pH, salinity and temperature. This specific strain utilizes both paraffin and anthracene as samples of aliphatic and polycyclic aromatic hydrocarbons. The ability of this bacterium to acquire all its energy and chemical requirements from Vacuum Distillation Residue (VR, as a net sample of problematic hydrocarbons in refineries, was studied. SARA test ASTM D4124-01 revealed 65.5% decrease in asphaltenic, 22.1% in aliphatics and 30.3% in Aromatics content of the VR in MSM medium. Further results with 0.9% saline showed 55% decrease in asphaltene content and 2.1% Aromatics respectively.Conclusion:Remarkable abilities of this microorganism propose its application in an ecofriendly technology to upgrade heavy crude oils.

  4. A Low Dimensional Approximation For Competence In Bacillus Subtilis.

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    Nguyen, An; Prugel-Bennett, Adam; Dasmahapatra, Srinandan

    2016-01-01

    The behaviour of a high dimensional stochastic system described by a chemical master equation (CME) depends on many parameters, rendering explicit simulation an inefficient method for exploring the properties of such models. Capturing their behaviour by low-dimensional models makes analysis of system behaviour tractable. In this paper, we present low dimensional models for the noise-induced excitable dynamics in Bacillus subtilis, whereby a key protein ComK, which drives a complex chain of reactions leading to bacterial competence, gets expressed rapidly in large quantities (competent state) before subsiding to low levels of expression (vegetative state). These rapid reactions suggest the application of an adiabatic approximation of the dynamics of the regulatory model that, however, lead to competence durations that are incorrect by a factor of 2. We apply a modified version of an iterative functional procedure that faithfully approximates the time-course of the trajectories in terms of a two-dimensional model involving proteins ComK and ComS. Furthermore, in order to describe the bimodal bivariate marginal probability distribution obtained from the Gillespie simulations of the CME, we introduce a tunable multiplicative noise term in a two-dimensional Langevin model whose stationary state is described by the time-independent solution of the corresponding Fokker-Planck equation. PMID:27045827

  5. Laundry detergent compatibility of the alkaline protease from Bacillus cereus.

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    Banik, Rathindra Mohan; Prakash, Monika

    2004-01-01

    The endogenous protease activity in various commercially available laundry detergents of international companies was studied. The maximum protease activity was found at 50 degrees C in pH range 10.5-11.0 in all the tested laundry detergents. The endogenous protease activity in the tested detergents retained up to 70% on incubation at 40 degrees C for 1 h, whereas less than 30% activity was only found on incubation at 50 degrees C for 1 h. The alkaline protease from an alkalophilic strain of Bacillus cereus was studied for its compatibility in commercial detergents. The cell free fermented broth from shake flask culture of the organism showed maximum activity at pH 10.5 and 50 degrees C. The protease from B. cereus showed much higher residual activity (more than 80%) on incubation with laundry detergents at 50 degrees C for 1 h or longer. The protease enzyme from B. cereus was found to be superior over the endogenous proteases present in the tested commercial laundry detergents in comparison to the enzyme stability during the washing at higher temperature, e.g., 40-50 degrees C. PMID:15293947

  6. Complexity in regulation of tryptophan biosynthesis in Bacillus subtilis.

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    Gollnick, Paul; Babitzke, Paul; Antson, Alfred; Yanofsky, Charles

    2005-01-01

    Bacillus subtilis uses novel regulatory mechanisms in controlling expression of its genes of tryptophan synthesis and transport. These mechanisms respond to changes in the intracellular concentrations of free tryptophan and uncharged tRNA(Trp). The major B. subtilis protein that regulates tryptophan biosynthesis is the tryptophan-activated RNA-binding attenuation protein, TRAP. TRAP is a ring-shaped molecule composed of 11 identical subunits. Active TRAP binds to unique RNA segments containing multiple trinucleotide (NAG) repeats. Binding regulates both transcription termination and translation in the trp operon, and translation of other coding regions relevant to tryptophan metabolism. When there is a deficiency of charged tRNA(Trp), B. subtilis forms an anti-TRAP protein, AT. AT antagonizes TRAP function, thereby increasing expression of all the genes regulated by TRAP. Thus B. subtilis and Escherichia coli respond to identical regulatory signals, tryptophan and uncharged tRNA(Trp), yet they employ different mechanisms in regulating trp gene expression. PMID:16285852

  7. Initiation of decay of Bacillus subtilis trp leader RNA.

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    Deikus, Gintaras; Bechhofer, David H

    2007-07-13

    Transcription termination in the leader region of the Bacillus subtilis trp operon is regulated by binding of the 11-mer TRAP complex to nascent trp RNA, which results in formation of a terminator structure. Rapid decay of trp leader RNA, which is required to release the TRAP complex and maintain a sufficient supply of free TRAP, is mediated by polynucleotide phosphorylase (PNPase). Using purified B. subtilis PNPase, we showed that, when TRAP was present, PNPase binding to the 3' end of trp leader RNA and PNPase digestion of trp leader RNA from the 3' end were inefficient. These results suggested that initiation of trp leader RNA may begin with an endonuclease cleavage upstream of the transcription terminator structure. Such cleavage was observed in vivo. Mutagenesis of nucleotides at the cleavage site abolished processing and resulted in a 4-fold increase in trp leader RNA half-life. This is the first mapping of a decay-initiating endonuclease cleavage site on a native B. subtilis RNA. PMID:17507374

  8. Hepatitis in Disseminated Bacillus Calmette-Guérin Infection

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    Markus U Göttke

    2000-01-01

    Full Text Available Local immunotherapy with an attenuated live strain of Mycobacterium bovis, bacillus Calmette-Guérin (BCG, is an effective and frequently used treatment for in situ transitional cell carcinoma (TCC of the bladder. Success rates are high, and serious side effects are infrequent but can affect every organ system. A 79-year-old patient with recently diagnosed TCC who was treated with intravesical BCG for a recurrence after initial surgical treatment is reported. After unsuccessful attempts at bladder catheterization with the creation of a false passage for his third treatment, BCG was instilled via a suprapubic catheter the same day and again a week later. Two weeks after the third BCG instillation, the patient presented with profound lethargy and weakness to the point of not being able to get up out of a chair. He was febrile, anorexic, icteric and had hepatosplenomegaly. Disseminated BCG infection was suspected on the basis of history, clinical examination and a liver biopsy that showed noncaseating granulomatous hepatitis. Empirical treatment was started with antituberculous combination therapy. A short course of an oral corticosteroid was given. Clinical improvement was marked and sustained so that the patient could be discharged home for the full six-month course of his treatment. Disseminated BCG infection with granulomatous hepatitis can be severe and life-threatening in cases where a large intravascular inoculum of BCG may have been given inadvertently.

  9. Properties of an immobilized lipase of Bacillus coagulans BTS-1.

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    Kanwari, S S; Srivastava, M; Chimni, S S; Ghazi, I A; Kaushal, R K; Joshi, G K

    2004-01-01

    Lipase (EC 3.1.1.3) is a tri-acylglycerol ester hydrolase, catalysing the hydrolysis of tri-, di-, and mono-acylglycerols to glycerol and fatty acids. To study the effect of adsorption of a lipase obtained from Bacillus coagulans BTS-1, its lipase was immobilized on native and activated (alkylated) matrices, i.e. silica and celite. The effect of pH, temperature, detergents, substrates, alcohols, organic solvent etc. on the stability of the immobilized enzyme was evaluated. The gluteraldahyde or formaldehyde (at 1% and 2% concentration, v/v) activated matrix was exposed to the Tris buffered lipase. The enzyme was adsorbed/entrapped more rapidly on to the activated silica than on the activated celite. The immobilized lipase showed optimal activity at 50 degrees C following one-hour incubation. The lipase was specifically more hydrolytic to the medium C-length ester (p-nitro phenyl caprylate than p-nitro phenyl laurate). The immobilization/entrapment enhanced the stability of the lipase at a relatively higher temperature (50 degrees C) and also promoted enzyme activity at an acidic pH (pH 5.5). Moreover, the immobilized lipase was quite resistant to the denaturing effect of SDS.

  10. Heterologous expression, biochemical characterization, and overproduction of alkaline α-amylase from Bacillus alcalophilus in Bacillus subtilis

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    Li Jianghua

    2011-10-01

    Full Text Available Abstract Background Alkaline α-amylases have potential applications for hydrolyzing starch under high pH conditions in the starch and textile industries and as ingredients in detergents for automatic dishwashers and laundries. While the alkaline α-amylase gains increased industrial interest, the yield of alkaline α-amylases from wild-type microbes is low, and the combination of genetic engineering and process optimization is necessary to achieve the overproduction of alkaline α-amylase. Results The alkaline α-amylase gene from Bacillus alcalophilus JN21 (CCTCC NO. M 2011229 was cloned and expressed in Bacillus subtilis strain WB600 with vector pMA5. The recombinant alkaline α-amylase was stable at pH from 7.0 to 11.0 and temperature below 40°C. The optimum pH and temperature of alkaline α-amylase was 9.0 and 50°C, respectively. Using soluble starch as the substrate, the Km and Vmax of alkaline α-amylase were 9.64 g/L and 0.80 g/(L·min, respectively. The effects of medium compositions (starch, peptone, and soybean meal and temperature on the recombinant production of alkaline α-amylase in B. subtilis were investigated. Under the optimal conditions (starch concentration 0.6% (w/v, peptone concentration 1.45% (w/v, soybean meal concentration 1.3% (w/v, and temperature 37°C, the highest yield of alkaline α-amylase reached 415 U/mL. The yield of alkaline α-amylase in a 3-L fermentor reached 441 U/mL, which was 79 times that of native alkaline α-amylase from B. alcalophilus JN21. Conclusions This is the first report concerning the heterologous expression of alkaline α-amylase in B. subtilis, and the obtained results make it feasible to achieve the industrial production of alkaline α-amylase with the recombinant B. subtilis.

  11. A Study on Effect of different culture media on amylase enzyme production by a native strain of Bacillus subtilis

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    ziba Akbari

    2015-12-01

    Full Text Available Introduction: Amylases are among the most important enzymes and have great significance in present-day biotechnology. Amylase with commercial applications is mainly derived from the genus Bacillus. The main purpose of this study is identification and isolatation amylase enzyme producer Bacillus, determining the amylase enzyme activity and affecting a number of culture medium on amylase enzyme production. Materials and methods: Soil, water and wastewater samples were collected from agricultural area, choghakhor lake in chahar mahal e bakhtiari province and from food factory in Esfahan. Bacillus isolates were screened for amylolytic properties by starch hydrolysis test on starch agar plate. Amylase producing Bacillus were identified biochemical tests and molecular experiments. Amylase enzyme activity of isolates was measured using di-nitro salicylic acid (DNS method. Enzyme production was studied in variose medium culture TSB, NB, Yeast extract, molases and milk medium. Results: The enzyme amylase-producing strains, one sample showed was the highest amylase activity. The Bacillus has been detected as a member of Bacillus subtilis according to Bergey's Manual of Systematic Bacteriology and molecular recognition. The enzyme activity of Bacillus subtilis was measured 7/21 (U/ml in production media. Trough medium culture maximum amylase production for Bacillus subtilis was achieved in molases medium. Discussion and conclusion: In this study, Bacillus subtilis strains isolated from wastewater of a significant amount of enzyme producing 7/21 (U/ml as indicated. Among the medium-amylase from Bacillus subtilis highest enzyme activity was observed in beet molasses. According to this study, the use of Bacillus strains is an efficient way to achieve the amylase enzyme.

  12. Involvement of Colonizing Bacillus Isolates in Glucovanillin Hydrolysis during the Curing of Vanilla planifolia Andrews.

    Science.gov (United States)

    Chen, Yonggan; Gu, Fenglin; Li, Jihua; He, Shuzhen; Xu, Fei; Fang, Yiming

    2015-08-01

    Vanilla beans were analyzed using biochemical methods, which revealed that glucovanillin disperses from the inner part to the outer part of the vanilla bean during the curing process and is simultaneously hydrolyzed by β-d-glucosidase. Enzymatic hydrolysis was found to occur on the surface of the vanilla beans. Transcripts of the β-d-glucosidase gene (bgl) of colonizing microorganisms were detected. The results directly indicate that colonizing microorganisms are involved in glucovanillin hydrolysis. Phylogenetic analysis based on 16S rRNA gene sequences showed that the colonizing microorganisms mainly belonged to the Bacillus genus. bgl was detected in all the isolates and presented clustering similar to that of the isolate taxonomy. Furthermore, inoculation of green fluorescent protein-tagged isolates showed that the Bacillus isolates can colonize vanilla beans. Glucovanillin was metabolized as the sole source of carbon in a culture of the isolates within 24 h. These isolates presented unique glucovanillin degradation capabilities. Vanillin was the major volatile compound in the culture. Other compounds, such as α-cubebene, β-pinene, and guaiacol, were detected in some isolate cultures. Colonizing Bacillus isolates were found to hydrolyze glucovanillin in culture, indirectly demonstrating the involvement of colonizing Bacillus isolates in glucovanillin hydrolysis during the vanilla curing process. Based on these results, we conclude that colonizing Bacillus isolates produce β-d-glucosidase, which mediates glucovanillin hydrolysis and influences flavor formation.

  13. Bacillus lonarensis sp. nov., an alkalitolerant bacterium isolated from a soda lake.

    Science.gov (United States)

    Reddy, Sultanpuram Vishnuvardhan; Thirumala, Mothe; Farooq, Mohammed; Sasikala, Chintalapati; Ramana, Chintalapati Venkata

    2015-01-01

    A novel Gram-stain-positive, rod-shaped, motile and endospore-forming novel bacterial strain 25nlg(T) was isolated from Lonar soda lake, in India. Based on the 16S rRNA gene sequence analysis, it was identified as a member of Firmicutes, being most closely related to Bacillus patagoniensis PAT 05(T) (96.6 %) and other members in the genus Bacillus (APL2) and three unknown lipids (L2-4). The predominant isoprenoid quinone was MK-7. iso-C15:0 (41.7 %) was the predominant fatty acid, and significant proportions of anteiso-C15:0 (20.8 %), C12:0 (5.5 %), anteiso-C17:0 (4.9 %), iso-C17:0 (4.5 %) were also detected in the strain 25nlg(T). The DNA G+C content of the strain 25nlg(T) was 40.5 mol%. The results of molecular, physiological and biochemical tests allowed a clear phenotypic differentiation of strain 25nlg(T) from all other members of the genus Bacillus. Strain 25nlg(T) represents a novel member of the genus Bacillus, for which the name Bacillus lonarensis sp. nov. is proposed. The type strain is 25nlg(T) (=KCTC 33413(T) = LMG 27974(T) = CGMCC = 1.12817(T)). PMID:25294189

  14. Involvement of Colonizing Bacillus Isolates in Glucovanillin Hydrolysis during the Curing of Vanilla planifolia Andrews.

    Science.gov (United States)

    Chen, Yonggan; Gu, Fenglin; Li, Jihua; He, Shuzhen; Xu, Fei; Fang, Yiming

    2015-08-01

    Vanilla beans were analyzed using biochemical methods, which revealed that glucovanillin disperses from the inner part to the outer part of the vanilla bean during the curing process and is simultaneously hydrolyzed by β-d-glucosidase. Enzymatic hydrolysis was found to occur on the surface of the vanilla beans. Transcripts of the β-d-glucosidase gene (bgl) of colonizing microorganisms were detected. The results directly indicate that colonizing microorganisms are involved in glucovanillin hydrolysis. Phylogenetic analysis based on 16S rRNA gene sequences showed that the colonizing microorganisms mainly belonged to the Bacillus genus. bgl was detected in all the isolates and presented clustering similar to that of the isolate taxonomy. Furthermore, inoculation of green fluorescent protein-tagged isolates showed that the Bacillus isolates can colonize vanilla beans. Glucovanillin was metabolized as the sole source of carbon in a culture of the isolates within 24 h. These isolates presented unique glucovanillin degradation capabilities. Vanillin was the major volatile compound in the culture. Other compounds, such as α-cubebene, β-pinene, and guaiacol, were detected in some isolate cultures. Colonizing Bacillus isolates were found to hydrolyze glucovanillin in culture, indirectly demonstrating the involvement of colonizing Bacillus isolates in glucovanillin hydrolysis during the vanilla curing process. Based on these results, we conclude that colonizing Bacillus isolates produce β-d-glucosidase, which mediates glucovanillin hydrolysis and influences flavor formation. PMID:25979899

  15. Bacillus 'next generation' diagnostics: Moving from detection towards sub-typing and risk related strain profiling

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    Monika eEhling-Schulz

    2013-02-01

    Full Text Available The highly heterogeneous genus Bacillus comprises the largest species group of endospore forming bacteria. Because of their ubiquitous nature, Bacillus spores can enter food production at several stages resulting in significant economic losses and posing a potential risk to consumers due the capacity of certain Bacillus strains for toxin production. In the past, food microbiological diagnostics was focused on the determination of species using conventional culture based methods, which are still widely used. However, due to the extreme intraspecies diversity found in the genus Bacillus, DNA based identification and typing methods are gaining increasing importance in routine diagnostics. Several studies showed that certain characteristics are rather strain dependent than species specific. Therefore, the challenge for current and future Bacillus diagnostics is not only the efficient and accurate identification on species level but also the development of rapid methods to identify strains with specific characteristics (such as stress resistance or spoilage potential, trace contamination sources, and last but not least discriminate potential hazardous strains from non-toxic strains.

  16. An exogenous surfactant-producing Bacillus subtilis facilitates indigenous microbial enhanced oil recovery

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    Peike eGao

    2016-02-01

    Full Text Available This study used an exogenous lipopeptide-producing Bacillus subtilis to strengthen the indigenous microbial enhanced oil recovery (IMEOR process in a water-flooded reservoir in the laboratory. The microbial processes and driving mechanisms were investigated in terms of the changes in oil properties and the interplay between the exogenous Bacillus subtilis and indigenous microbial populations. The exogenous Bacillus subtilis is a lipopeptide producer, with a short growth cycle and no oil-degrading ability. The Bacillus subtilis facilitates the IMEOR process through improving oil emulsification and accelerating microbial growth with oil as the carbon source. Microbial community studies using quantitative PCR and high-throughput sequencing revealed that the exogenous Bacillus subtilis could live together with reservoir microbial populations, and did not exert an observable inhibitory effect on the indigenous microbial populations during nutrient stimulation. Core-flooding tests showed that the combined exogenous and indigenous microbial flooding increased oil displacement efficiency by 16.71%, compared with 7.59% in the control where only nutrients were added, demonstrating the application potential in enhanced oil recovery in water-flooded reservoirs, in particular, for reservoirs where IMEOR treatment cannot effectively improve oil recovery.

  17. Mosquitocidal properties of Bacillus species isolated from mangroves of Vellar estuary, Southeast coast of India.

    Science.gov (United States)

    Balakrishnan, S; Indira, K; Srinivasan, M

    2015-09-01

    Samples collected from the mangroves of Vellar estuary yielded a mosquitocidal bacterium, whose secondary metabolites exhibited mosquito larvicidal and pupicidal activity. The bacterium was isolated using standard microbiological methods and identified using classical biochemical tests. The mosquitocidal bacterium was identified as Bacillus subtilis, Bacillus thuringiensis, Bacillus sphaericus and Bacillus cereus. Mosquitocidal metabolite(s) was separated from the culture supernatant of the bacterium and its efficacy was against the larval and pupal stages of two different species of mosquitoes and determined in terms of LC50 and LC90. Mosquito larvicidal activity in terms of LC50 against Anopheleus stephensi and Aedes aegypti was 4.374 and 7.406 μl/ml and its pupicidal activity was 4.928 and 9.865 μl/ml, respectively. The present study proved that the mosquitocidal properties of the Bacillus species isolated from mangroves of Vellar estuary was evaluated as target species of mosquito vectors. This is an ideal eco-friendly approach for the vector control programs.

  18. Decontamination Efficacy and Skin Toxicity of Two Decontaminants against Bacillus anthracis.

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    Chad W Stratilo

    Full Text Available Decontamination of bacterial endospores such as Bacillus anthracis has traditionally required the use of harsh or caustic chemicals. The aim of this study was to evaluate the efficacy of a chlorine dioxide decontaminant in killing Bacillus anthracis spores in solution and on a human skin simulant (porcine cadaver skin, compared to that of commonly used sodium hypochlorite or soapy water decontamination procedures. In addition, the relative toxicities of these decontaminants were compared in human skin keratinocyte primary cultures. The chlorine dioxide decontaminant was similarly effective to sodium hypochlorite in reducing spore numbers of Bacillus anthracis Ames in liquid suspension after a 10 minute exposure. After five minutes, the chlorine dioxide product was significantly more efficacious. Decontamination of isolated swine skin contaminated with Bacillus anthracis Sterne with the chlorine dioxide product resulted in no viable spores sampled. The toxicity of the chlorine dioxide decontaminant was up to two orders of magnitude less than that of sodium hypochlorite in human skin keratinocyte cultures. In summary, the chlorine dioxide based decontaminant efficiently killed Bacillus anthracis spores in liquid suspension, as well as on isolated swine skin, and was less toxic than sodium hypochlorite in cultures of human skin keratinocytes.

  19. Antagonizing Aspergillus parasiticus and Promoting Peanut Growth of Bacillus Isolated from Peanut Geocarposphere Soil

    Institute of Scientific and Technical Information of China (English)

    XIAO Wei; YAN Pei-sheng; WU Han-qi; LIN Feng

    2014-01-01

    A set of 38 Bacillus strains isolated from peanut geocarposphere soil were screened as potential biological control agent anti-Aspergillus parasiticus. Tip-culture method with rapid and quantitative characteristics was used to determine anti-A. parasiticus activity and the process of isolation could be accelerated with this method. 12 out of 38 Bacillus strains showed high anti-alfatoxin production activity. These 12 Bacillus strains were selected to identify the characteristics of promoting peanuts germination rate. Pot experiment in greenhouse was carried out by using these strains which can promote peanuts germination. Phytohormones in the fermentation broth were also detected as another important reference factor to select the isolates as biological control agent with PGPR features. These Bacillus strains isolated from peanut geocarposphere soil not only had high ability anti-Aspergillus parasiticus, but also promoted peanut growth. Therefore, these Bacillus strains were well adapted to peanut production in the ifeld as biological control agent with plant growth promoting rhizobacteria (PGPR) features.

  20. Plasmid Mediated Antibiotic and Heavy Metal Resistance in Bacillus Strains Isolated From Soils in Rize, Turkey

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    Elif SEVİM

    2015-09-01

    Full Text Available Fifteen Bacillus strains which were isolated from soil samples were examined for resistance to 17 different antibiotics (ampicillin, methicillin, erythromycin, norfloxacin, cephalotine, gentamycin, ciprofloxacin, streptomycin, tobramycin, chloramphenicol, trimethoprim-sulfamethoxazole, tetracycline, vancomycin, oxacilin, neomycin, kanamycin and, novabiocin and to 10 different heavy metals (copper, lead, cobalt, chrome, iron, mercury, zinc, nickel, manganese and, cadmium and for the presence of plasmid DNA. A total of eleven strains (67% were resistant to at least one antibiotic. The most common resistance was observed against methicillin and oxacillin. The most resistance strains were found as Bacillus sp. B3 and Bacillus sp. B11. High heavy metal resistance against copper, chromium, zinc, iron and nickel was detected, but mercury and cobalt resistance was not detected, except for 3 strains (B3, B11, and B12 which showed mercury resistance. It has been determined that seven Bacillus strains have plasmids. The isolated plasmids were transformed into the Bacillus subtilis W168 and it was shown that heavy metal and antibiotic resistance determinants were carried on these plasmids. These results showed that there was a correlation between plasmid content and resistance for both antibiotic and heavy metal resistance