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Sample records for bacillus mojavensis strain

  1. 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...

  2. DEVELOPMENT OF IMPROVED ANAEROBIC GROWTH OF BACILLUS MOJAVENSIS STRAIN JF-2 FOR THE PURPOSE OF IMPROVED ANAEROBIC BIOSURFACTANT PRODUCTION FOR ENHANCED OIL RECOVERY

    Energy Technology Data Exchange (ETDEWEB)

    M.J. McInerney; M. Folmsbee; D. Nagle

    2004-05-31

    Our work focuses on the use of microorganisms to recover petroleum hydrocarbons that remain entrapped after current recovery technologies reach their economic limit. Capillary forces between the hydrocarbon and aqueous phases are largely responsible for trapping the hydrocarbons in the pores of the rock and large reductions in the interfacial tension between the hydrocarbon and aqueous phases are needed for hydrocarbon mobilization (1-3, 10, 11). Microorganisms produce a variety of biosurfactants (4), several of which generate the ultra low interfacial tensions needed for hydrocarbon mobilization (4, 5, 8). In particular, the lipopeptide biosurfactant produced by Bacillus mojavensis strain JF-2 reduces the interfacial tension between hydrocarbon and aqueous phases to very low levels (<0.016 mN/m) (8) (9). B. mojavensis JF-2 grows under the environmental conditions found in many oil reservoirs, i. e., anaerobic, NaCl concentrations up to 80 g l{sup -1}, and temperatures up to 45 C (6, 7), making it ideally suited for in situ applications. However, anaerobic growth of B. mojavensis JF-2 was inconsistent and difficult to replicate, which limited its use for in situ applications. Our initial studies revealed that enzymatic digests, such as Proteose Peptone, were required for anaerobic growth of Bacillus mojavensis JF-2. Subsequent purification of the growth-enhancing factor in Proteose Peptone resulted in the identification of the growth-enhancing factor as DNA or deoxyribonucleosides. The addition of salmon sperm DNA, herring sperm DNA, E. coli DNA or synthetic DNA (single or double stranded) to Medium E all supported anaerobic growth of JF-2. Further, we found that JF-2 required all four deoxyribonucleosides (deoxyadeonosine, deoxyguanosine, deoxycytidine and thymidine) for growth under strict anaerobic conditions. The requirement for the deoxyribonucleosides did not occur under aerobic growth conditions. DNA was not used as a sole energy source; sucrose was required

  3. 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.

  4. 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)

    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-41617**T and Bacillus malacitensis NRRL B-41618**T. Compara...

  5. Tolerance and bioaccumulation of U(VI) by Bacillus mojavensis and its solid phase preconcentration by Bacillus mojavensis immobilized multiwalled carbon nanotube.

    Science.gov (United States)

    Özdemir, Sadin; Oduncu, M Kadir; Kilinc, Ersin; Soylak, Mustafa

    2017-02-01

    In this study, uranium(VI) tolerance and bioaccumulation were investigated by using thermo -tolerant Bacillus mojavensis. The level of U(VI) was measured by UV-VIS spectrophotometry. The minimum inhibition concentration (MIC) value of U(VI) was experimented. Bacterial growth was not affected in the presence of 1.0 and 2.5 mg/L U(VI) at 36 h and the growth was partially affected in the presence of 5 mg/L U(VI) at 24 h. What was obtained from this study is that there was diversity in the various periods of the growth phases of metal bioaccumulation capacity, which was shown by B. mojavensis. The maximum bioaccumulation capacities were found to be 12.8, 22.7, and 48.2 mg/g dried bacteria, at 24th hours at concentration of 1.0, 2.5 and 5 mg/L U(VI), respectively. In addition to these, U(VI) has been preconcentrated on B. mojavensis immobilized MWCNT. Several factors such as pH, flow rate of solution, amount of biosorbent and support materials, eluent type, concentration and volume, the matrix interference effect on retention have been studied, and extraction conditions were optimized. Preconcentration factor was achieved as 60. Under the optimized conditions, the limit of detection (LOD) and quantification (LOQ) were calculated as 0.74 and 2.47 μg/L. The biosorption capacity of immobilized B. mojavensis was calculated for U(VI) as 25.8 mg/g. The results demonstrated that the immobilized biosorbent column could be reused at least 30 cycles of biosorption and desorption with the higher than 95% recovery. FT-IR and SEM analysis were performed to understand the surface properties of B. mojavensis.

  6. He said, she said: mRNA sequencing identifies specificity in metabolic response to Bacillus mojavensis lipopeptides in Fusarium verticillioides

    Science.gov (United States)

    Fusarium verticillioides is a mycotoxigenic fungus capable of both pathogenic and asymptomatic endophytic lifestyles in maize; such intimate association renders efficient chemical control cost-prohibitive. Bacillus mojavensis RRC101 is a maize endophyte demonstrating both in vitro antagonism of F. v...

  7. 一株抗马铃薯坏疽病莫海威芽孢杆菌(Bacillus mojavensis ZA1)培养条件的优化%Optimization of culture conditions of a biocontrol agent Bacillus mojavensis ZA1 to antagonize Phoma foveata

    Institute of Scientific and Technical Information of China (English)

    冯中红; 畅涛; 杨成德; 薛莉; 李旭; 杨小利

    2016-01-01

    为了提高生防菌株莫海威芽孢杆菌(Bacillus mojavensis ZA1)液体发酵的生物量,利用单因素试验设计与响应曲面试验设计相结合的方法对 ZA1摇瓶发酵条件进行了优化。结果表明,ZA1的最佳培养基配比为氯化铵14.25 g、玉米粉19 g、马铃薯237 g、水1000 mL,最佳发酵条件为 pH 7.7、培养温度28℃、转速180 r/min 及发酵时间36 h,ZA1优化后活菌数为4.12×1010 CFU/mL。通过中心组合试验设计确定 ZA1在10 L 发酵罐中的最佳溶氧量为60%和转速为180 r/min;最优条件下,发酵 ZA1的放罐时间确定为36 h,活菌数达到1.59×1011 CFU/mL。该结果为利用 ZA1开发生防制剂奠定了基础。%This work aimed to identify optimum fermentation shaker-flask culture conditions for Bacillus mo-javensis strain ZA1.Various single culture factors were combined according to a Box-Behnken design.The op-timum ratio of ingredients for the culture medium for ZA1was found to be:14.25 g ammonium chloride,19 g corn flour,and 237 g potato,in 1000 mL distilled water.The optimum cultivation conditions were pH 7.7, temperature 28℃,and shaken at 180 r/min for 36 h.Under these conditions,the viable count of B .mojavens-is strain ZA1 was 4.12×10 10 colony-forming units per mL (CFU/mL).Further testing using the central com-posite design method in a 10 L fermentation tank found the optimum for dissolved oxygen level and shaker rota-tion speed to be 60% saturation and 180 r/min respectively.Under these conditions,when fermentation time was 36 h in the fermentation tank,the viable count of B .mojavensis strain ZA1 reached 1.59×10 11 CFU/mL. The results of this study lay a solid foundation for the development of methodology for large scale production of biocontrol agents.

  8. Production and characterization of Iturinic lipopeptides as antifungal agents and biosurfactants produced by a marine pinctada martensii-derived Bacillus mojavensis B0621A.

    Science.gov (United States)

    Ma, Zongwang; Hu, Jiangchun

    2014-06-01

    Bacillus mojavensis B0621A was isolated from a pearl oyster Pinctada martensii collected from South China Sea. While screening for cyclic lipopeptides potentially useful as lead compounds for biological control against soil-bone fungal plant pathogens, three lipopeptides were isolated and purified from the fermentation broth of B. mojavensis B0621A via vacuum flash chromatography coupled with reversed-phase high performance liquid chromatography (RP-HPLC). The structural characterization and identification of these cyclic lipopeptides were performed by tandem mass spectrometry (MS/MS) combined with gas chromatography-mass spectrometry (GC-MS) analysis as well as chemical degradation. These lipopeptides were finally characterized as homologues of mojavensins, which contained identical amino acids back bones of asparagine1, tyrosine2, asparagine3, glutamine4, proline5, asparagine6, and asparagine7 and differed from each other by their saturated β-amino fatty acid chain residues, namely, iso-C14 mojavensin, iso-C16 mojavensin, and anteiso-C17 mojavensin, respectively. All lipopeptide isomers, especially iso-C16 mojavensin and anteiso-C17 mojavensin, displayed moderate antagonism and dose-dependent activity against several formae speciales of Fusarium oxysporum and presented surface tension activities. These properties demonstrated that the lipopeptides produced by B. mojavensis B0621A may be useful as biological control agent to fungal plant pathogens.

  9. Assessment of pectinase production by Bacillus mojavensis I4 using an economical substrate and its potential application in oil sesame extraction.

    Science.gov (United States)

    Ghazala, Imen; Sayari, Nadhem; Romdhane, Molka Ben; Ellouz-Chaabouni, Semia; Haddar, Anissa

    2015-12-01

    Carrot (Daucus carota) peels, local agricultural waste product, is rich in lignocellulolytic material, including pectin which can act as an inducer of pectinase production. Pectinolytic enzymes production by Bacillus mojavensis I4 was studied in liquid state fermentation using carrot peel as a substrate. Medium composition and culture conditions for the pectinase production by I4 were optimized using two statistical methods: Taguchi design was applied to find the key ingredients and conditions for the best yield of enzyme production and The Box-Behnken design was used to optimize the value of the four significant variables: carrot peels powder, NH4Cl, inoculum size and incubation time. The optimal conditions for higher production of pectinase were carrot peels powder 6.5 %, NH4Cl 0.3 %, inoculum level 3 % and cultivation time 32 h. Under these conditions, the pectinase experimental yield (64.8 U/ml) closely matched the yield predicted by the statistical model (63.55 U/ml) with R (2) = 0.963. The best pectinase activity was observed at the temperature of 60 °C and at pH 8.0. The enzyme retained more than 90 % of its activity after 24 h at pH ranging from 6.0 to 10.0. The enzyme preserved more than 85 % of its initial activity after 60 min of pre-incubation at 30-40 °C and more than 67 % at 50 °C. The extracellular juice of I4 was applied in the process of sesame seeds oil extraction. An improvement of 3 % on the oil yield was obtained. The findings demonstrated that the B. mojavensis I4 has a promising potential for future use in a wide range of industrial and biotechnological applications.

  10. Characterization of Bacillus spp. strains for use as probiotic additives in pig feed

    DEFF Research Database (Denmark)

    Larsen, Nadja; Thorsen, Line; Kpikpi, Elmer Nayra

    2014-01-01

    for use as probiotic additives in pig feed. A total of 245 bacterial isolates derived from African fermented food, feces and soil were identified by 16S rRNA gene sequencing and screened for antimicrobial activity and growth in the presence of antibiotics, bile salts and at pH 4.0. Thirty-three Bacillus....... Isolates of B. amyloliquefaciens, B. subtilis and B. mojavensis, showed the best overall characteristics and, therefore, potential for usage as probiotic additives in feed. A large number of taxonomically diverse strains made it possible to reveal species and subspecies-specific trends, contributing to our...

  11. Draft Genome Sequences of Three Alkaliphilic Bacillus Strains, Bacillus wakoensis JCM 9140T, Bacillus akibai JCM 9157T, and Bacillus hemicellulosilyticus JCM 9152T

    OpenAIRE

    Yuki, Masahiro; Oshima, Kenshiro; Suda, Wataru; OSHIDA, Yumi; Kitamura, Keiko; Iida, Toshiya; Hattori, Masahira; Ohkuma, Moriya

    2014-01-01

    Here, we report the draft genome sequences of the type strains of three cellulolytic or hemicellulolytic alkaliphilic Bacillus species: Bacillus wakoensis, Bacillus akibai, and Bacillus hemicellulosilyticus. The genome information for these three strains will be useful for studies of alkaliphilic Bacillus species, their evolution, and biotechnological applications for their enzymes.

  12. Specific identification of Bacillus anthracis strains

    Science.gov (United States)

    Krishnamurthy, Thaiya; Deshpande, Samir; Hewel, Johannes; Liu, Hongbin; Wick, Charles H.; Yates, John R., III

    2007-01-01

    Accurate identification of human pathogens is the initial vital step in treating the civilian terrorism victims and military personnel afflicted in biological threat situations. We have applied a powerful multi-dimensional protein identification technology (MudPIT) along with newly generated software termed Profiler to identify the sequences of specific proteins observed for few strains of Bacillus anthracis, a human pathogen. Software termed Profiler was created to initially screen the MudPIT data of B. anthracis strains and establish the observed proteins specific for its strains. A database was also generated using Profiler containing marker proteins of B. anthracis and its strains, which in turn could be used for detecting the organism and its corresponding strains in samples. Analysis of the unknowns by our methodology, combining MudPIT and Profiler, led to the accurate identification of the anthracis strains present in samples. Thus, a new approach for the identification of B. anthracis strains in unknown samples, based on the molecular mass and sequences of marker proteins, has been ascertained.

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

    Science.gov (United States)

    Romero, Diego; Pérez-García, Alejandro; Veening, Jan-Willem; de Vicente, Antonio; Kuipers, Oscar P

    2006-09-01

    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 for molecular genetic analysis of interesting Bacillus strains, which are hard to transform by conventional methods.

  14. beta-Amylase production by some Bacillus cereus, Bacillus megaterium and Bacillus polymyxa [correction of polymaxa] strains.

    Science.gov (United States)

    Niziołek, S

    1997-01-01

    The production of extracellular beta-amylase by some Bacillus cereus, Bacillus megaterium and Bacillus polymyxa [corrected] strains was investigated, and the maximal yields of the enzyme were 3.6; 9.3 and 20.4 U/mL of the culture fluid, respectively (U, 1 mumol of maltose equivalent per min at 30 degrees C). Several cultivation media were used for beta-amylase production. Bacillus cereus and some strains of Bacillus megaterium gave good yields of beta-amylase only in medium with the addition of nutrient broth. However, beta-amylase produced during growth in protein rich medium (nutrient broth) was highly unstable, probably due to inactivation by proteolytic enzymes co-existing in the culture fluid. Bacillus polymyxa [corrected] strains can produce good yields of beta-amylase on a semi-synthetic medium consisting of inorganic salts, potato starch and inexpensive soybean extract instead of costly peptone and meat extract. The most potential beta-amylase producer was the strain Bacillus polymyxa [corrected] NCIB 8524. The tested Bacillus megaterium and Bacillus polymyxa [corrected] strains were apparently differentiated by temperature cultivation (30 and 37 degrees C) suitable for beta-amylase amylase yield.

  15. Identification of Bacillus strains for biological control of catfish pathogens.

    Science.gov (United States)

    Ran, Chao; Carrias, Abel; Williams, Malachi A; Capps, Nancy; Dan, Bui C T; Newton, Joseph C; Kloepper, Joseph W; 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 Edwardsiella tarda, Streptococcus iniae, Yersinia ruckeri, Flavobacterium columnare, and/or the oomycete Saprolegnia ferax. Survival of the 21 Bacillus strains in the intestine of catfish was determined as Bacillus CFU/g of intestinal tissue of catfish after feeding Bacillus spore-supplemented feed for seven days followed by normal feed for three days. Five Bacillus strains that showed good antimicrobial activity and intestinal survival were incorporated into feed in spore form at a dose of 8×10(7) CFU/g and fed to channel catfish for 14 days before they were challenged by E. ictaluri in replicate. Two Bacillus subtilis strains conferred significant benefit in reducing catfish mortality (Pbiological control in vivo was also investigated in terms of whether the strains contain plasmids or express resistance to clinically important antibiotics. The Bacillus strains identified from this study have good potential to mediate disease control as probiotic feed additives for catfish aquaculture.

  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. Identification of Bacillus strains for biological control of catfish pathogens.

    Directory of Open Access Journals (Sweden)

    Chao Ran

    Full Text Available 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 Edwardsiella tarda, Streptococcus iniae, Yersinia ruckeri, Flavobacterium columnare, and/or the oomycete Saprolegnia ferax. Survival of the 21 Bacillus strains in the intestine of catfish was determined as Bacillus CFU/g of intestinal tissue of catfish after feeding Bacillus spore-supplemented feed for seven days followed by normal feed for three days. Five Bacillus strains that showed good antimicrobial activity and intestinal survival were incorporated into feed in spore form at a dose of 8×10(7 CFU/g and fed to channel catfish for 14 days before they were challenged by E. ictaluri in replicate. Two Bacillus subtilis strains conferred significant benefit in reducing catfish mortality (P<0.05. A similar challenge experiment conducted in Vietnam with four of the five Bacillus strains also showed protective effects against E. ictaluri in striped catfish. Safety of the four strains exhibiting the strongest biological control in vivo was also investigated in terms of whether the strains contain plasmids or express resistance to clinically important antibiotics. The Bacillus strains identified from this study have good potential to mediate disease control as probiotic feed additives for catfish aquaculture.

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

    OpenAIRE

    Romero, Diego; Perez-Garcia, Alejandro; Veening, Jan-Willem; Vicente, Antonio; Oscar P. Kuipers; 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 for molecular genetic analysis of interesting Bacillus strains, which are hard to transform by conventional methods. (c) 2006 Elsevier B.V. All rights reserved.

  19. Construction of acetoin high-producing Bacillus subtilis strain

    Directory of Open Access Journals (Sweden)

    Yanjun Tian

    2016-07-01

    Full Text Available This paper describes the construction and selection of a high-producing mutant, Bacillus subtilis HB-32, with enhanced acetoin yield and productivity. The mutant was obtained by the protoplast fusion of a Bacillus subtilis mutant TH-49 (Val− producing acetoin and Bacillus licheniformis AD-30 producing α-acetolactate decarboxylase, with the fusogen polyethylene glycol and after the regeneration and selection, etc. of the fusant. The acetoin production reached 49.64 g/L, which is an increase of 61.8% compared to that of B. subtilis strain TH-49. Random amplified polymorphic DNA analysis was performed to determine the mutagenic and protoplast fusion effects and the genomic changes in the acetoin high-producing strain compared to the parent strains at the molecular level. The constructed strain was shown to be promising for large-scale acetoin production. Future studies should focus on the application of the mutant strain in practice.

  20. 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

  1. 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.

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

    Science.gov (United States)

    Zwick, Michael E; Kiley, Maureen P; Stewart, Andrew C; Mateczun, Alfred; Read, Timothy D

    2008-07-02

    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.

  3. Classification of Strains of Bacillus sphaericus by Different Statistical Methods

    OpenAIRE

    MERCAN, Nazime

    2003-01-01

    In this study, the sensitivies of 21 strains of Bacillus sphaericus to 20 different antibiotics were determined with the antibiotic disc susceptibility test. By measuring the diameters of the inhibition zones in terms of millimetres, the obtained values were analysed using the SPSS for Windows package and classifications were made. With these analyses, 21 strains were clustered into 5 groups. In our study, strains IAB 59 and 2297 were placed into a separate class. In addition, those antibioti...

  4. 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.

  5. 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.

  6. 77 FR 19109 - Bacillus Pumilus Strain GHA 180; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2012-03-30

    ... AGENCY 40 CFR Part 180 Bacillus Pumilus Strain GHA 180; Exemption From the Requirement of a Tolerance... an exemption from the requirement of a tolerance for residues of Bacillus pumilus strain GHA 180 in... permissible level for residues of Bacillus pumilus strain GHA 180. DATES: This regulation is effective...

  7. 77 FR 2910 - Bacillus Amyloliquefaciens Strain D747; Exemption From the Requirement of a Tolerance; Technical...

    Science.gov (United States)

    2012-01-20

    ... AGENCY 40 CFR Part 180 Bacillus Amyloliquefaciens Strain D747; Exemption From the Requirement of a... establishment of an exemption from the requirement of a tolerance for residues of Bacillus amyloliquefaciens strain D747 (formerly known as Bacillus subtilis variant amyloliquefaciens strain D747). This document...

  8. 77 FR 745 - Bacillus Amyloliquefaciens Strain D747; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2012-01-06

    ... AGENCY 40 CFR Part 180 Bacillus Amyloliquefaciens Strain D747; Exemption From the Requirement of a... establishes an exemption from the requirement of a tolerance for residues of the Bacillus amyloliquefaciens strain D747 (formerly known as Bacillus subtilis variant amyloliquefaciens strain D747) in or on all...

  9. 78 FR 35147 - Bacillus pumilus Strain BU F-33; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2013-06-12

    ... AGENCY 40 CFR Part 180 Bacillus pumilus Strain BU F-33; Exemption From the Requirement of a Tolerance... an exemption from the requirement of a tolerance for residues of Bacillus pumilus strain BU F-33 in... residues of Bacillus pumilus strain BU F-33 under the FFDCA. DATES: This regulation is effective June...

  10. Complete Genome Sequence of Bacillus thuringiensis Strain 407 Cry-

    OpenAIRE

    Poehlein, Anja; Liesegang, Heiko

    2013-01-01

    Bacillus thuringiensis is an insect pathogen that has been used widely as a biopesticide. Here, we report the genome sequence of strain 407 Cry-, which is used to study the genetic determinants of pathogenicity. The genome consists of a 5.5-Mb chromosome and nine plasmids, including a novel 502-kb megaplasmid.

  11. Bacillus nakamurai sp. nov., a black pigment producing strain

    Science.gov (United States)

    Two isolates of a Gram-positive, strictly aerobic, motile, rod-shaped, endospore-forming bacterium were identified during a survey of the Bacillus diversity of the Agriculture Research Service Culture Collection. These strains were originally isolated from soil and have a phenotype of producing a da...

  12. 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

  13. Genetic diversity among Bacillus anthracis, Bacillus cereus and Bacillus thuringiensis strains using repetitive element polymorphism-PCR.

    Science.gov (United States)

    Brumlik, Michael J; Bielawska-Drózd, Agata; Zakowska, Dorota; Liang, Xudong; Spalletta, Ronald A; Patra, Guy; Delvecchio, Vito G

    2004-01-01

    Repetitive element polymorphism-PCR (REP-PCR) is one of the tools that has been used to elucidate genetic diversity of related microorganisms. Using the MB1 primer, REP-PCR fingerprints from 110 Bacillus strains within the "B. cereus group" have identified eighteen distinct categories, while other more distantly related bacterial species fell within six additional categories. All Bacillus anthracis strains tested were found to be monomorphic by fluorophore-enhanced REP-PCR (FERP) fingerprinting using the MB1 primer. In contrast, other non- B. anthracis isolates displayed a high degree of polymorphism. Dendrogramic analysis revealed that the non- B. anthracis strains possessing the Ba813 chromosomal marker were divided into two clusters. One of the clusters shared identity with the B. cereus strains examined.

  14. Antimicrobial susceptibility of Bacillus anthracis strains from Hungary.

    Science.gov (United States)

    Kreizinger, Zsuzsa; Sulyok, Kinga Mária; Makrai, László; Rónai, Zsuzsanna; Fodor, László; Jánosi, Szilárd; Gyuranecz, Miklós

    2016-06-01

    The susceptibility of 29 Bacillus anthracis strains, collected in Hungary between 1933 and 2014, was tested to 10 antibiotics with commercially available minimum inhibitory concentration (MIC) test strips. All strains were susceptible to amoxicillin, ciprofloxacin, clindamycin, doxycycline, gentamicin, penicillin, rifampicin, and vancomycin. Intermediate susceptibility to erythromycin and cefotaxime was detected in 17.2% (5/29) and 58.6% (17/29) of the strains, respectively. Correlations were not observed between the isolation date, location, host species, genotype, and antibiotic susceptibility profile of strains.

  15. 77 FR 73934 - Bacillus subtilis Strain QST 713 Variant Soil; Amendment to an Exemption From the Requirement of...

    Science.gov (United States)

    2012-12-12

    ... AGENCY 40 CFR Part 180 Bacillus subtilis Strain QST 713 Variant Soil; Amendment to an Exemption From the Requirement of a Tolerance for Bacillus subtilis Strain QST 713 To Include Residues of Bacillus subtilis... Bacillus subtilis strain QST 713 in or on all food commodities by including residues of Bacillus...

  16. 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.

  17. 77 FR 1633 - Bacillus Subtilis Strain CX-9060; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2012-01-11

    ... AGENCY 40 CFR Part 180 Bacillus Subtilis Strain CX-9060; Exemption From the Requirement of a Tolerance... an exemption from the requirement of a tolerance for residues of the microbial pesticide Bacillus... eliminates the need to establish a maximum permissible level for residues of Bacillus subtilis strain...

  18. 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.

  19. Molecular characterization of Bacillus thuringiensis strains from Argentina.

    Science.gov (United States)

    Franco-Rivera, Alejandro; Benintende, Graciela; Cozzi, Jorge; Baizabal-Aguirre, Victor Manuel; Valdez-Alarcón, Juan José; López-Meza, Joel Edmundo

    2004-07-01

    Bacillus thuringiensis INTA 7-3, INTA 51-3, INTA Mo9-5 and INTA Mo14-4 strains were obtained from Argentina and characterized by determination of serotype, toxicity, plasmid composition, insecticidal gene content ( cry and vip ) and the cloning of the single- vip3A gene of the INTA Mo9-5 strain. The serotype analysis identified the serovars tohokuensis and darmstadiensis for the INTA 51-3 and INTA Mo14-4 strains, respectively, whereas the INTA Mo9-5 strain was classified as "autoagglutinated". In contrast to the plasmid patterns of INTA 7-3, INTA 51-3 and INTA Mo9-5 (which were similar to B. thuringiensis HD-1 strain), strain INTA Mo14-4 showed a unique plasmid array. PCR analysis of the four strains revealed the presence of cry genes and vip3A genes. Interestingly, it was found that B. thuringiensis 4Q7 strain, which is a plasmid cured strain, contained vip3A genes indicating the presence of these insecticidal genes in the chromosome. Bioassays towards various lepidopteran species revealed that B. thuringiensis INTA Mo9-5 and INTA 7-3 strains were highly active. In particular, the mean LC(50) obtained against A. gemmatalis larvae with the INTA Mo9-5 and INTA 7-3 strains were 7 (5.7-8.6) and 6.7 (5.6-8.0) ppm, respectively. The INTA Mo14-4 strain was non-toxic and strain INTA 51-3 showed only a weak larvicidal activity.

  20. Antibiotic Sensitivity of Bacillus clausii Strains in Commercial Preparation

    Science.gov (United States)

    Abbrescia, A.; Palese, L.L.; Papa, S.; Gaballo, A.; Alifano, P.; Sardanelli, Anna M.

    2014-01-01

    Recently has been acknowledged the healthy use of Bacillus and related bacteria as probiotics. A mixture reported to contain four probiotic strains of Bacillus clausii is marketed as an OTC (Over The Counter) medicinal supplement for human use. Their poliantibiotic resistant property, useful for restoring the gut microbiota during antibiotic treatment, raises the question about the risk of resistance transfer. In order to better assess the risk-benefit ratio it is important to always monitoring the pattern and stability of resistance spectra in these bacteria. In this work, we have extensively redefined the antibiotic susceptibility profile of these four probiotic strains. Resistance phenotype has been determined by screening a large number of antibiotics, including natural products (such as penicillin, vancomycin and erythromycin), and completely synthetic molecules (such as fluoroquinolones). Extensive comparison with a wild type strain belonging to the normal intestinal microbiome was carried out. The molecular basis of some resistances was determined. Observed antibiotic resistances were correlated with previous and new data in safety evaluations of these strains for human use.

  1. Environment driven cereulide production by emetic strains of Bacillus cereus.

    Science.gov (United States)

    Apetroaie-Constantin, Camelia; Shaheen, Ranad; Andrup, Lars; Smidt, Lasse; Rita, Hannu; Salkinoja-Salonen, Mirja

    2008-09-30

    The impacts of growth media and temperature on production of cereulide, the emetic toxin of Bacillus cereus, were measured for seven well characterised strains selected for diversity of biochemical and genetic properties and sources of origin. All strains carried cereulide synthase gene, ces, on a megaplasmid of ca. 200 kb and all grew up to 48-50 degrees C, but produced cereulide only up to 39 degrees C. On tryptic soy agar five strains, originating from foods, food poisonings and environment, produced highest amounts of cereulide at 23 to 28 degrees C, whereas two strains, from human faeces, produced cereulide similarly from 23 to 39 degrees C, with no clear temperature trend. These two strains differed from the others also by producing more cereulide on tryptic soy agar if supplemented with 5 vol.% of blood, whereas the other five strains produced similarly, independent on the presence of blood. On oatmeal agar only one strain produced major amounts of cereulide. On skim milk agar, raw milk agar, and MacConkey agar most strains grew well but produced only low amounts of cereulide. Three media components, the ratio [K+]:[Na+], contents of glycine and [Na+], appeared of significance for predicting cereulide production. Increase of [K+]:[Na+] (focal variable) predicted (P cereus in a complex manner. The relevance of the findings to production of cereulide in the gut and to the safety of amino acids as additives in foods containing live toxinogenic organisms is discussed.

  2. 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.

  3. Physiologic and genetic characterization of Bacillus sphaericus native strains

    Directory of Open Access Journals (Sweden)

    Jenny Dussen Garzón

    2012-06-01

    Full Text Available Eighteen pathogenic native strains of Bacillus sphaericus that were pathogens to mosquito larvae were isolated from different Colombia regions. The objective of this study was to evaluate at physiological and molecular level pathogenic strains and to compare them with the reference 2362 one. Cellular growth, sporulation percentage, pathogenic activity against third instar larvae of Culex quinquejasciaius, the presence of toxigenic proteins, the size of native plasmids and the genetic polymorphism among pathogenic and non pathogenic isolations was evaluated. The evaluated strains of Bs presented a latency stage ranging from 2 to 3 h and one logarithmic phase of 7 h; the sporulation in BHI was lower than 1% to 40 h of incubation, in NYSM medium was obtained ten times more production of biomass and spores, 26% of the population showed sporulation percentage higher than 90%; the isolations were classified in three groups of pathogenicity with [Formula] and [Formula]. Punctual mutations on the genes which encoding for native toxins were detected and was found a exclusive protein of 30 kDa in pathogenic native strains. The isolations of Bs presented a plasmid of 118Kb that was not related to the toxicity; pathogenic strains are a homogenous group with a similarity between 90 to 100%, whereas non-pathogenic ones are genetically heterogeneous group and conform a cluster aside. The native strains have a great potential in biological control of mosquito larvae that transmitting diseases such as dengue, malaria, encephalitis and filariasis among others.

  4. Complete Genome Sequence of Bacillus subtilis subsp. subtilis Strain 3NA

    OpenAIRE

    Reuß, Daniel R.; Schuldes, Jörg; Daniel, Rolf; Altenbuchner, Josef

    2015-01-01

    Bacillus subtilis 3NA reaches high cell densities during fed-batch fermentation and is an interesting target for further optimization as a production strain. Here, we announce the full genome of B. subtilis 3NA. The presence of specific Bacillus subtilis 168 and W23 genetic features suggests that 3NA is a hybrid of these strains.

  5. Antifungal activity and gene expression of lipopeptide antibiotics in strains of genus Bacillus

    OpenAIRE

    Grabova A. Yu.; Dragovoz I. V.; Zelena L. B.; Tkachuk D. M.; Avdeeva L. V.

    2016-01-01

    Aim. To research the antifungal activity and gene expression of lipopeptide antibiotics in strains of genus Bacillus. Methods. Deferred antagonism method, PCR, qRT-PCR, MALDI-TOF mass spectrometry. Results. It was revealed that Bacillus sp. strains C6 and Lg37s out of five tested strains had the highest antifungal activity. Based on the molecular genetic methods, it was shown that the expression of genes of lipopeptide antibiotics, related to the fengycin family, occurred in all these strains...

  6. Genome Sequences of Bacillus thuringiensis Serovar kurstaki Strain BP865 and B. thuringiensis Serovar aizawai Strain HD-133

    Science.gov (United States)

    Jeong, Haeyoung

    2017-01-01

    ABSTRACT We report the draft genome sequences of two insecticidal strains against lepidopteran pests, Bacillus thuringiensis serovar kurstaki strain BP865, an isolate from the South Korean phylloplane, and strain HD-133, a reference strain of B. thuringiensis serovar aizawai. PMID:28153898

  7. Comparative proteome analysis of two antagonist Bacillus subtilis strains.

    Science.gov (United States)

    Zhang, C X; Zhao, X; Han, F; Yang, M F; Chen, H; Chida, T; Shen, S H

    2009-04-01

    Natural wild-type strains of Bacillus subtilis are extensively used in agriculture as biocontrol agents for plants. This study examined two antagonist B. subtilis strains, KB-1111 and KB-1122, and the results illustrated that KB-1122 was a more potent inhibitor of the indicator pathogen than KB- 1111. Thus, to investigate the intrinsic differences between the two antagonist strains under normal culture conditions, samples of KB-1111 and KB-1122 were analyzed using MALDI-TOF-MS. The main differences were related to 20 abundant intracellular and 17 extracellular proteins. When searching the NCBI database, a number of the differentially expressed proteins were identified, including 11 cellular proteins and 10 secretory proteins. Among these proteins, class III stress-response-related ATPase, aconitate hydratase, alpha-amylase precursor, and a secretory protein, endo-1, 4-beta-glucanase, were differentially expressed by the two strains. These results are useful to comprehend the intrinsic differences between the antagonism of KB-1111 and KB-1122.

  8. Antifungal activity and gene expression of lipopeptide antibiotics in strains of genus Bacillus

    Directory of Open Access Journals (Sweden)

    Grabova A. Yu.

    2016-02-01

    Full Text Available Aim. To research the antifungal activity and gene expression of lipopeptide antibiotics in strains of genus Bacillus. Methods. Deferred antagonism method, PCR, qRT-PCR, MALDI-TOF mass spectrometry. Results. It was revealed that Bacillus sp. strains C6 and Lg37s out of five tested strains had the highest antifungal activity. Based on the molecular genetic methods, it was shown that the expression of genes of lipopeptide antibiotics, related to the fengycin family, occurred in all these strains. At the same time, the gene expression of cyclolipopeptide iturin was found in the Bacillus sp. strains C6 and Lg37s. It was determined that Bacillus sp. C6 strain had the highest level of expression of the fengycin operon`s genes, whereas the lowest level was observed in Bacillus sp. C10 strain. By means of MALDI-TOF mass spectrometry, the presence of fengycins in the cell-free cultural fluid of Bacillus sp. C6 strain was detected. Conclusion. The direct correlation between the level of antifungal activity and the fengycin synthetases expression has not been disclosed. A higher level of antagonism detected for two Bacillus strains is more likely associated with the expression and subsequent synthesis of fengycin and iturin.

  9. 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.

  10. 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.

  11. 40 CFR 180.1209 - Bacillus subtilis strain QST 713; 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 strain QST 713... RESIDUES IN FOOD Exemptions From Tolerances § 180.1209 Bacillus subtilis strain QST 713; exemption from the... the microbial pesticide Bacillus subtilis strain QST 713 when used in or on all food commodities....

  12. 40 CFR 180.1255 - Bacillus pumilus strain QST 2808; 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 pumilus strain QST 2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1255 Bacillus pumilus strain QST 2808; exemption from the... the microbial pesticide Bacillus pumilus strain QST 2808 when used in or on all...

  13. 40 CFR 180.1226 - Bacillus pumilus strain QST2808; temporary 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 pumilus strain QST2808... RESIDUES IN FOOD Exemptions From Tolerances § 180.1226 Bacillus pumilus strain QST2808; temporary exemption... established for residues of the microbial pesticide Bacillus pumilus strain QST2808 when used in or on...

  14. [Features of Bacillus subtilis IMB B-7023 and its streptomycin-resistant strain].

    Science.gov (United States)

    Roĭ, A A; Iatsenko, I P; Gordienko, A S; Kurdish, I K

    2011-01-01

    Features of phosphate-mobilizing bacteria Bacillus subtilis IMB B-7023 and its streptomycin-resistant strain were investigated. While cultivated in medium with glucose and glycerophosphate, the growth rate of the antibiotic-marked strain was approximately similar to this parameter for Bacillus subtilis IMB B-7023 but cell sizes were 1.3-fold less. Both strains significantly stimulated the germinating of plant seeds, attached to their roots, and insignificantly differed in antagonistic activity toward phytopathogens and quantitative content of cell fatty acids and phosphatase activity. Streptomycin-resistant strain may be used for monitoring of Bacillus subtilis introduced to agroecosystem.

  15. [BACILLUS STRAINS'S SCREENING--ACTIVE ANTAGONISTS OF BACTERIAL AND FUNGAL PHYTOPATHOGENS].

    Science.gov (United States)

    Grabova, A Yu; Dragovoz, I V; Kruchkova, L A; Pasichnik, L A; Avdeeva, L V

    2015-01-01

    Antagonistic activity 100 strains of Bacillus bacteria towards to museum and actual strains of phytopathogenic bacteria and fungy was defined. Relation between level of antagonistic activity to phytopathogenic bacteria and genus accessory of the last was shown. The medium level of antagonism to fungal phytopathogens at 30% of the studied strains of Bacillus bacteria was shown. 5 strains of Bacillus sp. with high and medium levels of antagonism to phytopathogens bacterial and fungy nature was selected and considered as perspective for creation of biological preparations for plant protection.

  16. 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.

  17. 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.

  18. Multilocus sequence typing reveals that Bacillus cereus strains isolated from clinical infections have distinct phylogenetic origins.

    Science.gov (United States)

    Barker, Margaret; Thakker, Bishan; Priest, Fergus G

    2005-04-01

    Eight strains of Bacillus cereus isolated from bacteremia and soft tissue infections were assigned to seven sequence types (STs) by multilocus sequence typing (MLST). Two strains from different locations had identical STs. The concatenated sequences of the seven STs were aligned with 65 concatenated sequences from reference STs and a neighbor-joining tree was constructed. Two strains were distantly related to all reference STs. Three strains were recovered in a clade that included Bacillus anthracis, B. cereus and rare Bacillus thuringiensis strains while the other three strains were assigned to two STs that were more closely affiliated to most of the B. thuringiensis STs. We conclude that invasive B. cereus strains do not form a single clone or clonal complex of highly virulent strains.

  19. 40 CFR 180.1181 - Bacillus cereus strain BPO1; 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 cereus strain BPO1; exemption... FOOD Exemptions From Tolerances § 180.1181 Bacillus cereus strain BPO1; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance for residues of the Bacillus...

  20. Isolation and characterisation of Bacillus spp. antagonistic to Vibrio parahaemolyticus for use as probiotics in aquaculture.

    Science.gov (United States)

    Liu, Xue-Fei; Li, Ya; Li, Jian-Rong; Cai, Lu-Yun; Li, Xiu-Xia; Chen, Jin-Ru; Lyu, Shu-Xia

    2015-05-01

    Acute gastroenteritis caused by pathogenic Vibrio parahaemolyticus is one of the major factors affecting the development of aquaculture and the safety of seafood. Using the antagonism of probiotics against pathogens is an alternative strategy to antibiotics and a common trend to control food-borne pathogenic bacteria. In this study, a total of 249 isolates were isolated from four types of seafood (Litopenaeus vannamei, Oratosquilla oratoria, Mactra veneriformis and Portunus trituberculatus) and coastal sediment from Liaodong Bay in the Bohai Sea, China with five different separation agars. The most isolates came from the sample of coastal sediment and on agar of 2216E, which accounted for 36.14 and 54.62 % respectively. Twenty-four among 249 isolates displayed direct antimicrobial activity to V. parahaemolyticus with spot inoculation. Sixteen active isolates were selected for extracellular antimicrobial activity using the Oxford cup method. Only strains of B16 and J7 showed extracellular antimicrobial activity and were identified as Bacillus pumilus and Bacillus mojavensis respectively based on the physiological identification and 16S rRNA sequence analysis. Both of the strains B16 and J7 exhibited extracellular hydrolytic enzyme activity and antagonism against more than one indicator bacteria in vitro, which indicates that the two strains have broad potential application as suitable probiotic candidates in aquaculture while B. mojavensis was first reported to inhibit pathogenic Vibrio spp. in vitro. There is no particular trait as to antagonism of B. pumilus B16 or B. mojavensis J7 to Gram-positive or Gram-negative indicator bacteria.

  1. [Joint cultivation of Bacillus subtilis and Escherichia coli strains promising for obtaining complex probiotic].

    Science.gov (United States)

    Tsaruk'ianova, I G; Osadchaia, A I

    2007-01-01

    The ability of joint cultivation of Bacillus subtilis UCM B-5007 and Escherichia coli M-17 in subsurface conditions has been studied. These strains are available for creation of a new complex probiotic. Symbiotic relationships between these microorganisms were proved. Bacillus subtilis and Escherichia coli strains use different growth "strategy". The most optimum ratio of cultures (1:1) for growth, biomass accumulation, and for antagonism to test-cultures has been chosen.

  2. 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.

  3. A novel neutral protease from thermophilic Bacillus strain HUTBS62

    Directory of Open Access Journals (Sweden)

    HAZEM AQEL

    2012-01-01

    Full Text Available A novel neutral highly thermostable protease was detected in the culture medium of thermophilic Bacillus strain HUTBS62 isolated from hot-spring located near to the Dead Sea, Jordan. The enzyme was purified by precipitation with 55-60% ammonium sulfate, gel filtration on Sephadex G-100 and DEAE ion exchange chromatography. The enzyme was purified 53-fold with 2% yield. The optimum pH and temperature for catalytic activity of protease was pH 6.8 and 80ºC, respectively, and 31% activity of protease remained even after heat treatment at 100ºC for 60 min. The relative activity of the enzyme was highly stable (90% at 50ºC for 2 h. The half-life of the enzyme at 90ºC, 80ºC and 70ºC was estimated to be 3, 4 and 6 h, respectively. The activation energy of denaturation of purified enzyme was 21.7 kJmol-1. Iron, sodium, calcium, and manganese increased protease activity. On the other hand, magnesium, cobalt and zinc variably decreased the residual activity. But cadmium and copper drastically inhibited the enzyme activity. The enzymatic activity was highly stable in the presence of 1 and 2 mM EDTA at pH 6.8 and 80ºC. The neutral protease therefore could be defined as a highly thermostable with new properties make the present enzyme applicable for many biotechnological purposes.

  4. Expression of the neutral protease gene from a thermophilic Bacillus sp BT1 strain in Bacillus subtilis and its natural host : Identification of a functional promoter

    NARCIS (Netherlands)

    Vecerek, B; Venema, G

    2000-01-01

    The expression of the neutral protease gene (npr) from the thermophilic Bacillus sp. BT1 strain was studied in its natural host and in mesophilic Bacillus subtilis. In the thermophilic BT1 strain, the transcription of the protease gene is initiated from its own promoter, just 5' to the gene. In cont

  5. Draft Genome Sequence of Bacillus tequilensis Strain FJAT-14262a

    OpenAIRE

    Chen, Qian-Qian; Liu, Bo; Liu, Guo-hong; Wang, Jie-ping; Che, Jian-Mei

    2015-01-01

    Bacillus tequilensis FJAT-14262a is a Gram-positive rod-shaped bacterium. Here, we report the 4,038,551-bp genome sequence of B. tequilensis FJAT-14262a, which will provide useful information for genomic taxonomy and phylogenomics of Bacillus.

  6. Draft Genome Sequence of Bacillus tequilensis Strain FJAT-14262a.

    Science.gov (United States)

    Chen, Qian-Qian; Liu, Bo; Liu, Guo-Hong; Wang, Jie-Ping; Che, Jian-Mei

    2015-11-12

    Bacillus tequilensis FJAT-14262a is a Gram-positive rod-shaped bacterium. Here, we report the 4,038,551-bp genome sequence of B. tequilensis FJAT-14262a, which will provide useful information for genomic taxonomy and phylogenomics of Bacillus.

  7. Differentiation of strains from the Bacillus cereus group by RFLP-PFGE genomic fingerprinting.

    Science.gov (United States)

    Otlewska, Anna; Oltuszak-Walczak, Elzbieta; Walczak, Piotr

    2013-11-01

    Bacillus mycoides, Bacillus pseudomycoides, Bacillus weihenstephanensis, Bacillus anthracis, Bacillus thuringiensis, and Bacillus cereus belong to the B. cereus group. The last three species are characterized by different phenotype features and pathogenicity spectrum, but it has been shown that these species are genetically closely related. The macrorestriction analysis of the genomic DNA with the NotI enzyme was used to generate polymorphism of restriction profiles for 39 food-borne isolates (B. cereus, B. mycoides) and seven reference strains (B. mycoides, B. thuringiensis, B. weihenstephanensis, and B. cereus). The PFGE method was applied to differentiate the examined strains of the B. cereus group. On the basis of the unweighted pair group method with the arithmetic mean method and Dice coefficient, the strains were divided into five clusters (types A-E), and the most numerous group was group A (25 strains). A total of 21 distinct pulsotypes were observed. The RFLP-PFGE analysis was successfully used for the differentiation and characterization of B. cereus and B. mycoides strains isolated from different food products.

  8. DnaJ sequences of Bacillus cereus strains isolated from outbreaks of hospital infection are highly similar to Bacillus anthracis.

    Science.gov (United States)

    Zhang, Jiwei; van Hung, Pham; Hayashi, Masahiro; Yoshida, Shigeru; Ohkusu, Kiyofumi; Ezaki, Takayuki

    2011-07-01

    Bacillus cereus is becoming an important nomosomial pathogen because of frequent isolation from blood cultures and from severe systemic infections. To differentiate highly pathogenic outbreak strain of B. cereus from other sources of the Bacillus cereus, we attempted to analyze their dnaJ sequences. Assays indicated that dnaJ sequence similarity of all of 52 blood culture isolates of B. cereus ranged from 92.8% to 100%. The distance between B. anthracis and B. cereus except six outbreak isolates ranged from 3.8% to 6.4%. The dnaJ sequences of six outbreak strains of B. cereus (GTC 02891, GTC 02896, GTC 02916, GTC 02917, GTC 03221, and GTC 03222) were closely related to those of B. anthracis (99.2%-99.5% sequence similarity). Ba813 sequences were only found in the six outbreak strains of B. cereus. The other pathogenic factors of B. anthracis were not found in these six outbreak strains, with the exception of GTC 02891 (cap-positive). The six outbreak strains formed clear β-hemolytic colonies on a sheep blood agar plate. Our findings suggest that outbreak strains of B. cereus isolated from blood cultures are likely to have the risk of causing serious infection, and dnaJ and Ba813 are important markers to identify such strains. Phylogenetic analysis of dnaJ and MLST revealed that the six outbreak strains of B. cereus are closely related to B. anthracis.

  9. Genotype Analysis of Bacillus anthracis Strains Circulating in Bangladesh.

    Science.gov (United States)

    Rume, Farzana Islam; Affuso, Alessia; Serrecchia, Luigina; Rondinone, Valeria; Manzulli, Viviana; Campese, Emanuele; Di Taranto, Pietro; Biswas, Paritosh Kumar; Ahsan, Chowdhury Rafiqul; Yasmin, Mahmuda; Fasanella, Antonio; Hugh-Jones, Martin

    2016-01-01

    In Bangladesh, anthrax, caused by the bacterium Bacillus anthracis, is considered an endemic disease affecting ruminants with sporadic zoonotic occurrences in humans. Due to the lack of knowledge about risks from an incorrect removal of infected carcasses, the disease is not properly monitored, and because of the socio-economic conditions, the situation is under-reported and under-diagnosed. For sensitive species, anthrax represents a fatal outcome with sudden death and sometimes bleeding from natural orifices. The most common source of infection for ruminants is ingestion of spores during grazing in contaminated pastures or through grass and water contaminated with anthrax spores. Domestic cattle, sheep and goats can also become infected through contaminated bone meal (used as feed) originating from anthrax-infected carcasses. The present investigation was conducted to isolate B. anthracis organisms from 169 samples (73 soil, 1 tissue, 4 bone and 91 bone meal samples) collected from 12 different districts of Bangladesh. The sampling was carried out from 2012 to 2015. Twelve samples resulted positive for B. anthracis. Biomolecular analyses were conducted starting from the Canonical Single Nucleotide Polymorphism (CanSNP) to analyze the phylogenetic origin of strains. The analysis of genotype, obtained through the Multiple Locus Variable Number Tandem Repeat Analysis (MLVA) with the analysis of 15 Variable Number Tandem Repeats (VNTR), demonstrated four different genotypes: two of them were previously identified in the district of Sirajganj. The sub-genotyping, conducted with Single Nucleotide Repeats analysis, revealed the presence of eight subgenotypes. The data of the present study concluded that there was no observed correlation between imported cattle feed and anthrax occurrence in Bangladesh and that the remarkable genetic variations of B. anthracis were found in the soil of numerous outbreaks in this country.

  10. Flavins mediate extracellular electron transfer in Gram-positive Bacillus megaterium strain LLD-1

    DEFF Research Database (Denmark)

    Xiao, Yong; Liu, Lidan; You, Lexing;

    electrochemically active strain of Bacillus megatherium strain LLD-1, and its extracellular electron transfer mechanism was demonstrated by cyclic voltammetry (CV), differential pulse voltammetry (DPV), HPLC, and chronoamperometric. The CV and DPV showed that a redox peaks ascribing to membrane proteins was found...

  11. Metabolic capacity of Bacillus cereus strains ATCC 14579 and ATCC 10987 interlinked with comparative genomics.

    NARCIS (Netherlands)

    Mols, M.; Been, M.W.H.J. de; Zwietering, M.H.; Moezelaar, R.; Abee, T.

    2007-01-01

    Bacillus cereus is an important food-borne pathogen and spoilage organism. In this study, numerous phenotypes and the genomes of B.?cereus strains ATCC 14579 and ATCC 10987 were analysed to compare their metabolic capacity and stress resistance potential. The growth performance of the two strains wa

  12. Discovery of secondary metabolites from Bacillus spp. biocontrol strains using genome mining and mass spectroscopy

    Science.gov (United States)

    Genome sequencing, data mining and mass spectrometry were used to identify secondary metabolites produced by several Bacillus spp. biocontrol strains. These biocontrol strains have shown promise in managing Fusarium head blight in wheat. Draft genomes were produced and screened in silico using genom...

  13. Genome Sequence of the Soviet/Russian Bacillus anthracis Vaccine Strain 55-VNIIVViM

    Science.gov (United States)

    Kotorashvili, Adam

    2016-01-01

    Bacillus anthracis strain 55-VNIIVViM is a live-attenuated nonencapsulated Soviet/Russian veterinary anthrax vaccine strain. We report here the genome of 55-VNIIVViM and confirm its phylogenetic placement in the global population structure of B. anthracis. PMID:28007853

  14. Development of an efficient electroporation method for rhizobacterial Bacillus mycoides strains

    NARCIS (Netherlands)

    Yi, Yanglei; Kuipers, Oscar P

    2016-01-01

    In order to develop a method for electroporation of environmental Bacillus mycoides strains, we optimized several conditions that affect the electroporation efficiency of this bacterium. By combining the optimized conditions, the electroporation efficiency of strain EC18 was improved to (1.3 ± 0.6)

  15. CHARACTERIZATION OF A NEW BACILLUS-STEAROTHERMOPHILUS ISOLATE - A HIGHLY THERMOSTABLE ALPHA-AMYLASE-PRODUCING STRAIN

    NARCIS (Netherlands)

    WIND, RD; BUITELAAR, RM; EGGINK, G; HUIZING, HJ; DIJKHUIZEN, L

    1994-01-01

    A novel strain of Bacillus stearothermophilus was isolated from samples of a potato-processing industry. Compared to known alpha-amylases from other B. stearothermophilus strains, the isolate was found to produce a highly thermostable alpha-amylase. The half-time of inactivation of this alpha-amylas

  16. Characterization of a new Bacillus stearothermophilus isolate : a highly thermostable α-amylase-producing strain

    NARCIS (Netherlands)

    Wind, R.D.; Buitelaar, R.M.; Eggink, G.; Huizing, H.J.; Dijkhuizen, L.

    1994-01-01

    A novel strain of Bacillus stearothermophilus was isolated from samples of a potato-processing industry. Compared to known α-amylases from other B. stearothermophilus strains, the isolate was found to produce a highly thermostable α-amylase. The half-time of inactivation of this α-amylase was 5.1 h

  17. Identification of "Bacillus cellulasensis" strain NIO-1130(T) as a member of Bacillus altitudinis and emendation of the latter.

    Science.gov (United States)

    Liu, Yang; Lai, Qiliang; Shao, Zongze

    2016-10-01

    In the study by Mawlankar et al. in Arch Microbiol 198:83-89 (2016), the phylogenetic position of strain "Bacillus cellulasensis" NIO-1130(T) based on 16S rRNA and gyrB genes was inconsistent. Therefore, the aim of this study is to re-determine its taxonomic status using diverse genotypic approaches including single gene analysis, multilocus sequence analysis, and genomic analyses. The reconstructed phylogenetic trees based on 16S rRNA gene and six concatenated genes showed that "B. cellulasensis" NIO-1130(T) (=NCIM 5461(T) = CCTCC AB 2011126(T)) revealed the closest genetic relationship with type strain Bacillus altitudinis 41KF2b(T), with 98.6-100 % similarities of 16S rRNA gene, gyrB, pycA, pyrE, mutL, aroE, trpB, and six concatenated housekeeping genes. The high similarities for gene(s) sequences between "B. cellulasensis" NIO-1130(T) and B. altitudinis 41KF2b(T) indicated that they should be conspecific. The DNA G+C content for strain NIO-1130(T) was determined to be 41.3 mol% and identical to that of B. altitudinis 41KF2b(T). Moreover, 88.4 % of digital DNA-DNA hybridization and 98.7 % of average nucleotide identity values between two strains were much higher than the standard criteria for delineation of bacterial species, suggesting that they belonged to the same species. Therefore, the data from the combined genotypic analyses suggest that "Bacillus cellulasensis" should be classified as a member of Bacillus altitudinis.

  18. Genome sequence of the aerobic bacterium Bacillus sp. strain FJAT-13831.

    Science.gov (United States)

    Liu, Guohong; Liu, Bo; Lin, Naiquan; Tang, Weiqi; Tang, Jianyang; Lin, Yingzhi

    2012-12-01

    Bacillus sp. strain FJAT-13831 was isolated from the no. 1 pit soil of Emperor Qin's Terracotta Warriors in Xi'an City, People's Republic of China. The isolate showed a close relationship to the Bacillus cereus group. The draft genome sequence of Bacillus sp. FJAT-13831 was 4,425,198 bp in size and consisted of 5,567 genes (protein-coding sequences [CDS]) with an average length of 782 bp and a G+C value of 36.36%.

  19. Plasmid Mediated Antibiotic and Heavy Metal Resistance in Bacillus Strains Isolated From Soils in Rize, Turkey

    Directory of Open Access Journals (Sweden)

    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

  20. Bacillus 'next generation' diagnostics: Moving from detection towards sub-typing and risk related strain profiling

    Directory of Open Access Journals (Sweden)

    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.

  1. Optimization of polyphosphate production by Bacillus megaterium strain G11

    Directory of Open Access Journals (Sweden)

    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

  2. The Genetically Remote Pathogenic Strain NVH391-98 of the Bacillus cereus Group Represents the Cluster of Thermophilic Strains

    Energy Technology Data Exchange (ETDEWEB)

    Auger, Sandrine; Galleron, Nathalie; Bidnenko, Elena; Ehrlich, S. Dusko; Lapidus, Alla; Sorokin, Alexei

    2007-10-02

    Bacteria of the Bacillus cereus group are known to cause food poisoning. A rare phylogenetically remote strain, NVH391-98, was recently characterized to encode a particularly efficient cytotoxin K presumably responsible for food poisoning. This pathogenic strain and its close relatives can be phenotypically distinguished from other strains of the B. cereus group by the inability to grow at temperatures below 17 degrees C and by the ability to grow at temperatures from 48 to 53 degrees C. A temperate phage, phBC391A2, residing in the genome of NVH391-98 allows us to distinguish the three known members of this thermophilic strain cluster.

  3. 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.

  4. Environmental regulation of alcohol metabolism in thermotolerant methylotrophic Bacillus strains

    NARCIS (Netherlands)

    Arfman, N.; Moezelaar, H.R.; Attwood, M.M.; Robinson, G.K.; Geel, M. van; Dijkhuizen, L.

    1992-01-01

    The thermotolerant methylotroph Bacillus sp. C1 possesses a novel NAD-dependent methanol dehydrogenase (MDH), with distinct structural and mechanistic properties. During growth on methanol and ethanol, MDH was responsible for the oxidation of both these substrates. MDH activity in cells grown on met

  5. Molecular and toxigenic characterization of Bacillus cereus and Bacillus thuringiensis strains isolated from commercial ground roasted coffee.

    Science.gov (United States)

    Chaves, Jeane Quintanilha; Cavados, Clara de Fátima Gomes; Vivoni, Adriana Marcos

    2012-03-01

    Thirty samples of roasted ground coffee beans from 10 different commercial brands were analyzed to investigate the occurrence and levels of Bacillus cereus and Bacillus thuringiensis strains. Strains were evaluated for their genetic diversity by repetitive element sequence polymorphism PCR (Rep-PCR) and for their toxigenic profiles, i.e., the presence of hblA, hblC, hblD, nheA, nheB, nheC, cytK, ces, and entFM. Survival and multiplication of B. cereus sensu lato in the ready-to-drink coffee was determined to evaluate this beverage as a possible vehicle for B. cereus infection. B. cereus was detected in 17 (56.7%) of the 30 samples, and B. thuringiensis was detected in 8 (26.7%) of the 30 samples. Five samples did not produce any characteristic growth. The most common gene, entFM, was detected in 23 strains (92%). The NHE complex (nheA, nheB, and nheC genes) was found in 19 strains (76%). The HBL complex (hblA, hblC, and hblD) was found in 16 strains (64%). All strains were negative for ces. The cytK gene was found in 16 strains (64%). The computer-assisted cluster analysis of Rep-PCR profiles using a clustering criterion of 80% similarity revealed four main clusters. Cluster 1 was the predominant and comprised three B. thuringiensis strains with 100% similarity, cluster 2 comprised two B. cereus strains (100% similarity), cluster 3 comprised two B. thuringiensis strains (90% similarity), and cluster 4 comprised one B. thuringiensis strain and one B. cereus strain (85% similarity). The cluster analysis of fingerprints generated by Rep-PCR revealed a high genetic diversity among the B. cereus strains, suggesting that the contamination could have originated from different sources. In our experiments, when sugar was added and the beverage was kept in thermic bottles there was a significant increase in B. cereus sensu lato levels, which may increase the risk of food poisoning. These results highlight the need for additional studies on this subject to better evaluate

  6. Inhibition of mycotoxin-producing fungi by Bacillus strains isolated from fish intestines.

    Science.gov (United States)

    Veras, Flávio Fonseca; Correa, Ana Paula Folmer; Welke, Juliane Elisa; Brandelli, Adriano

    2016-12-05

    Bacillus strains isolated from the aquatic environment of the Brazilian Amazon region were tested for their activity against mycotoxigenic fungi. All tested bacteria showed antifungal activity, inhibiting at least 7 indicator fungi. Four Bacillus strains showing promising antifungal results were subsequently evaluated for their activity in reducing mycelial growth rate, sporulation, spore germination percentage, and mycotoxin production. Bacillus sp. P1 and Bacillus sp. P11 had a remarkable antifungal effect on toxigenic fungi. Washed bacterial cell suspension of strains P1 and P11 (10(7)CFU/ml) reduced by >70% the fungal colony diameters, including a complete inhibition of ochratoxin A (OTA) producing Aspergillus spp. Significant reduction of growth rate, sporulation and spore germination were also observed. The bacteria influenced the production of mycotoxins, causing a reduction around 99 and 97% in AFB1 and OTA concentration, respectively. Chromatographic analysis revealed the presence of lipopeptides (iturin A and surfactin isomers) in butanol extracts of cell-free supernatants and cell pellets of strains P1 and P11. Furthermore, antifungal activity of these extracts was confirmed against A. flavus A12 and A. carbonarius ITAL293, producers of AFB1 and OTA, respectively. These bacterial strains could be promising biocontrol agents against toxigenic fungi.

  7. Production of lipopeptides among Bacillus strains showing growth inhibition of phytopathogenic fungi.

    Science.gov (United States)

    Velho, R V; Medina, L F C; Segalin, J; Brandelli, A

    2011-07-01

    The biological activity and the presence of genes sfp and ituD (surfactin and iturin A) among Bacillus strains isolated from the Amazon basin were determined. Bacillus spp. were tested for hemolytic activity and inhibition of fungal growth by agar plate assays in parallel with PCR for identification of sfp and ituD genes. All strains tested produced surface-active compounds, giving evidence by lysis of erythrocytes and emulsifying activity on mineral oil and soybean oil. These strains of Bacillus caused growth inhibition of several phytopathogenic fungi, including Fusarium spp., Aspergillus spp., and Bipolaris sorokiniana. The presence of genes ituD and sfp was confirmed by PCR and sequence analysis. The only exception was Bacillus sp. P34 that lacks sfp gene. Lipopeptides were isolated from culture supernatants and analyzed by mass spectrometry. Characteristic m/z peaks for surfactin and iturin were observed, and some strains also produced fengycin and bacillomycin. The remarkable antifungal activity showed by the strains could be associated with the co-production of three or more lipopeptide antibiotics. Screening for novel bacteria producing useful biosurfactants or biocontrol agents for agriculture is a topic of greatest importance to eliminate chemical pollutants.

  8. Antimicrobial activities of rhizobacterial strains of Pseudomonas and Bacillus strains isolated from rhizosphere soil of carnation (Dianthus caryophyllus cv. Sunrise).

    Science.gov (United States)

    Sharma, Sapna; Kaur, Mohinder

    2010-06-01

    Under the present study, an attempt was made to characterize rhizobacteria i.e. Pseudomonas and Bacillus species isolated from rhizosphere of carnation to evaluate their growth promoting effect on carnation so as to select and develop more efficient indigenous plant growth promoting and disease suppressing bioagents of specific soil type and specific plant type. Maximum strains of Pseudomonas and Bacillus sp. showed significant antimicrobial activities against most of the microorganisms tested. On the basis of in vitro antagonistic activities, the best strains were selected and used in field trial to study the influence of these strains on the growth of carnation. Results have shown marked effect on growth parameters and disease incidence has also been reduced significantly.

  9. Production of Enzymes from Agroindustrial Wastes by Biosurfactant-Producing Strains of Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Francisco Fábio Cavalcante Barros

    2013-01-01

    Full Text Available Bacteria in the genus Bacillus are the source of several enzymes of current industrial interest. Hydrolases, such as amylases, proteases, and lipases, are the main enzymes consumed worldwide and have applications in a wide range of products and industrial processes. Fermentation processes by Bacillus subtilis using cassava wastewater as a substrate are reported in the technical literature; however, the same combination of microorganisms and this culture medium is limited or nonexistent. In this paper, the amylase, protease, and lipase production of ten Bacillus subtilis strains previously identified as biosurfactant producers in cassava wastewater was evaluated. The LB1a and LB5a strains were selected for analysis using a synthetic medium and cassava wastewater and were identified as good enzyme producers, especially of amylases and proteases. In addition, the enzymatic activity results indicate that cassava wastewater was better than the synthetic medium for the induction of these enzymes.

  10. Biological characteristics of Bacillus thuringiensis strain Btll and identification of its cry-type genes

    Institute of Scientific and Technical Information of China (English)

    Tinghui LIU; Wei GUO; Weiming SUN; Yongxiang SUN

    2009-01-01

    A novel strain of Bacillus thuringiensis Bt11, isolated from soil samples in China, was classified and characterized in terms of its crystal proteins, cry genes content. The Bt11 strain showed high toxicity against Spodoptera exigua and Helicoverpa armigera neonates. Btll strain shares morphological and biochemical characteristics with the previously described Bacillus thuringiensis subsp. kurstaki. SDS-polyacrylamide gel electrophoresis revealed that crystals were composed of several polypeptides ranging from 20 to 130 kDa, of which the 35, 80, and 130 kDa proteins were the major components. PCR-RFLP with total DNA from strain Btll and specific primers for cryl, cry2, cry3, cry4/10, cry7, cry8, cry9, and cryll genes revealed that crylAa, crylAb, crylla, and cry9Ea genes were present.

  11. Complete Genome Sequence of Bacillus amyloliquefaciens Strain BH072, Isolated from Honey

    NARCIS (Netherlands)

    Zhao, Xin; de Jong, Anne; Zhou, Zhijiang; Kuipers, Oscar P

    2015-01-01

    The genome of Bacillus amyloliquefaciens strain BH072, isolated from a honey sample and showing strong antimicrobial activity against plant pathogens, is 4.07 Mb and harbors 3,785 coding sequences (CDS). Several gene clusters for nonribosomal synthesis of antimicrobial peptides and a complete gene c

  12. Genome Sequence of Bacillus thuringiensis Strain Btm27, an Egyptian Isolate Highly Toxic to Cotton Leafworm

    Science.gov (United States)

    Rusconi, Brigida; Chen, Yue; Koenig, Sara S. K.; El-Helow, Ehab R.

    2015-01-01

    Bacillus thuringiensis is a potent microbial control agent against insect pests. Here, we present the draft genome of the Egyptian strain Btm27 that shows high toxicity toward the cotton leafworm. The genome contains three insecticidal genes cry1Ac9, cry2Ab1, and vip3V that have been implicated in conferring toxicity toward lepidoptera. PMID:25977430

  13. Complete Genome Sequence of Bacillus thuringiensis Serovar Tolworthi Strain Pasteur Institute Standard

    Science.gov (United States)

    Kanda, Kohzo; Nakashima, Kaede

    2015-01-01

    The genome sequence of Bacillus thuringiensis serovar tolworthi strain Pasteur Institute Standard was determined. The genome consists of a 5.9-Mb chromosome and eight plasmids, one of which is linear. The second largest plasmid (293 kb) carries the genes encoding insecticidal proteins. PMID:26139717

  14. Study on Attenuation Characteristics of Biocontrol Strain Anti-8098A, Bacillus cereus, against Ralstoniasolanacearum

    Institute of Scientific and Technical Information of China (English)

    BoLIU; Ying-ZhiLIN; Yu-JingZHU; Ci-BinGE; YiCAO

    2004-01-01

    The present study dealt with the attenuation characteristics of bacterial-wilt-disease biocontrol strain Anti-8098A, Bacillus cereus, againstpathogeny Ralstonia solanacearum (RS). In order to distinguish the pathogenicity of RS, the attenuation index (radius of the center red ring/radius of the whole mycelium ring, on TTC culture medium) was established (Hayward, 1976), companying with the mortality of tomato

  15. Tryptophan provision by dietary supplementation of a Bacillus subtilis mutant strain in piglets

    DEFF Research Database (Denmark)

    Torres-Pitarch, A; Nielsen, B.; Canibe, Nuria

    2015-01-01

    Supplementing Bacillus (B.) subtilis mutants selected to overproduce a specific amino acid (AA) may be an alternative method to provide essential AA in pig diets. Two experiments on a B. subtilis strain selected to overproduce Trp were conducted using 8-kg pigs fed Trp-deficient diets for 20 d. B...

  16. Use of Long-Range Repetitive Element Polymorphism-PCR To Differentiate Bacillus anthracis Strains

    OpenAIRE

    Brumlik, Michael J.; Szymajda, Urszula; Zakowska, Dorota; Liang, Xudong; Redkar, Rajendra J.; Patra, Guy; Del Vecchio, Vito G.

    2001-01-01

    The genome of Bacillus anthracis is extremely monomorphic, and thus individual strains have often proven to be recalcitrant to differentiation at the molecular level. Long-range repetitive element polymorphism-PCR (LR REP-PCR) was used to differentiate various B. anthracis strains. A single PCR primer derived from a repetitive DNA element was able to amplify variable segments of a bacterial genome as large as 10 kb. We were able to characterize five genetically distinct groups by examining 10...

  17. Exoproteome analysis of a novel strain of Bacillus cereus implicated in disease resembling cutaneous anthrax.

    Science.gov (United States)

    Ghosh, Neha; Goel, Ajay Kumar; Alam, Syed Imteyaz

    2014-03-01

    Bacillus cereus belongs to B. cereus sensu lato group, shared by six other related species including Bacillus anthracis. B. anthracis is the causative agent for serious illness affecting a wide range of animals as well as humans and is a category A Biological and Toxin Warfare (BTW) agent. Recent studies indicate that a Bacillus species other than B. anthracis can cause anthrax-like disease and role of anthrax virulence plasmids (pXO1 and pXO2) on the pathogenicity of B. cereus has been documented. B. cereus strain TF5 was isolated from the tissue fluid of cutaneous anthrax-like skin lesions of a human patient from an anthrax endemic area in India. The strain harboured a PA gene, however, presence of pXO1 or pXO2-like plasmids could not be ascertained using reported primers. Abundant exoproteome of the strain in the early stationary phase was elucidated using a 2-DE MS approach and compared with that from a reference B. cereus strain. Analysis of proteins showing qualitative and quantitative differences between the two strains indicated an altered regulatory mechanism and putative role of S-layer protein and sphingomyelinase in the pathogenesis of strain TF5. Phylogenetic analysis of the S-layer protein indicated close affiliation of the strain with anthracis-like B. cereus strains such as B. cereus var. anthracis strain CI; whereas sphingomyelinase exhibited specific relationship with all the strains of B. anthracis apart from that with anthracis-like B. cereus strains.

  18. Physiological Properties and Salmonella Growth Inhibition of Probiotic Bacillus Strains Isolated from Environmental and Poultry Sources.

    Science.gov (United States)

    Menconi, Anita; Morgan, Marion J; Pumford, Neil R; Hargis, Billy M; Tellez, Guillermo

    2013-01-01

    The objective of the present study was to describe the physiological properties of seven potential probiotic strains of Bacillus spp. Isolates were characterized morphologically, biochemically, and by 16S rRNA sequence analyses for identification. Tolerance to acidic pH, high osmotic concentrations of NaCl, and bile salts were tested. Isolates were also evaluated for their ability to metabolize different carbohydrates sources. The antimicrobial sensitivity profiles were determined. Inhibition of gastrointestinal Salmonella colonization in an avian model was also evaluated. Five strains of Bacillus were tolerant to acidic conditions (pH 2.0) and all strains were tolerant to a high osmotic pressure (NaCl at 6.5%). Moreover, all strains were able to tolerate concentration of 0.037% bile salts after 24 h of incubation. Three strains were able to significantly reduce Salmonella Typhimurium levels in the crop and in the ceca of broiler-type chickens. Among the 12 antibiotics tested for antibiotic resistance, all strains were resistant to bacitracin and susceptible to gentamycin, neomycin, ormethoprim, triple sulfa, and spectinomycin. Bacterial spore formers have been shown to prevent gastrointestinal diseases in animals and humans. The results obtained in this study show important characteristics to be evaluated when selecting Bacillus spp. candidates to be used as probiotics.

  19. Physiological Properties and Salmonella Growth Inhibition of Probiotic Bacillus Strains Isolated from Environmental and Poultry Sources

    Directory of Open Access Journals (Sweden)

    Anita Menconi

    2013-01-01

    Full Text Available The objective of the present study was to describe the physiological properties of seven potential probiotic strains of Bacillus spp. Isolates were characterized morphologically, biochemically, and by 16S rRNA sequence analyses for identification. Tolerance to acidic pH, high osmotic concentrations of NaCl, and bile salts were tested. Isolates were also evaluated for their ability to metabolize different carbohydrates sources. The antimicrobial sensitivity profiles were determined. Inhibition of gastrointestinal Salmonella colonization in an avian model was also evaluated. Five strains of Bacillus were tolerant to acidic conditions (pH 2.0 and all strains were tolerant to a high osmotic pressure (NaCl at 6.5%. Moreover, all strains were able to tolerate concentration of 0.037% bile salts after 24 h of incubation. Three strains were able to significantly reduce Salmonella Typhimurium levels in the crop and in the ceca of broiler-type chickens. Among the 12 antibiotics tested for antibiotic resistance, all strains were resistant to bacitracin and susceptible to gentamycin, neomycin, ormethoprim, triple sulfa, and spectinomycin. Bacterial spore formers have been shown to prevent gastrointestinal diseases in animals and humans. The results obtained in this study show important characteristics to be evaluated when selecting Bacillus spp. candidates to be used as probiotics.

  20. Degradation of polyester polyurethane by a newly isolated soil bacterium, Bacillus subtilis strain MZA-75.

    Science.gov (United States)

    Shah, Ziaullah; Krumholz, Lee; Aktas, Deniz Fulya; Hasan, Fariha; Khattak, Mutiullah; Shah, Aamer Ali

    2013-11-01

    A polyurethane (PU) degrading bacterial strain MZA-75 was isolated from soil through enrichment technique. The bacterium was identified through 16S rRNA gene sequencing, the phylogenetic analysis indicated the strain MZA-75 belonged to genus Bacillus having maximum similarity with Bacillus subtilis strain JBE0016. The degradation of PU films by strain MZA-75 in mineral salt medium (MSM) was analyzed by scanning electron microscopy (SEM), fourier transform infra-red spectroscopy (FT-IR) and gel permeation chromatography (GPC). SEM revealed the appearance of widespread cracks on the surface. FTIR spectrum showed decrease in ester functional group. Increase in polydispersity index was observed in GPC, which indicates chain scission as a result of microbial treatment. CO2 evolution and cell growth increased when PU was used as carbon source in MSM in Sturm test. Increase in both cell associated and extracellular esterases was observed in the presence of PU indicated by p-Nitrophenyl acetate (pNPA) hydrolysis assay. Analysis of cell free supernatant by gas chromatography-mass spectrometry (GC-MS) revealed that 1,4-butanediol and adipic acid monomers were produced. Bacillus subtilis strain MZA-75 can degrade the soft segment of polyester polyurethane, unfortunately no information about the fate of hard segment could be obtained. Growth of strain MZA-75 in the presence of these metabolites indicated mineralization of ester hydrolysis products into CO2 and H2O.

  1. High-quality genome sequence and description of Bacillus ndiopicus strain FF3T sp. nov.

    Directory of Open Access Journals (Sweden)

    C.I. Lo

    2015-11-01

    Full Text Available Strain FF3T was isolated from the skin-flora of a 39-year-old healthy Senegalese man. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry did not allow any identification. This strain exhibited a 16S rRNA sequence similarity of 96.8% with Bacillus massiliensis, the phylogenetically closest species with standing nomenclature. Using a polyphasic study made of phenotypic and genomic analyses, strain FF3T was Gram-positive, aeroanaerobic and rod shaped and exhibited a genome of 4 068 720 bp with a G+C content of 37.03% that coded 3982 protein-coding and 67 RNA genes (including four rRNA operons. On the basis of these data, we propose the creation of Bacillus ndiopicus sp. nov.

  2. Bacillus sp. strain DJ-1, potent arsenic hypertolerant bacterium isolated from the industrial effluent of India.

    Science.gov (United States)

    Joshi, Dhaval N; Flora, S J S; Kalia, Kiran

    2009-07-30

    Arsenic hypertolerant bacterial cells were isolated from the common industrial effluent treatment plant, Vapi, India. Strain DJ-1 sustaining 400 mM, As (V) out of 16 bacterial strains was identified as Bacillus sp. strain DJ-1 through 16S rRNA ribotyping. The maximum arsenic accumulation of 9.8+/-0.5 mg g(-1) (dry weight) was observed during stationary phase of growth. Intracellular compartmentalization has shown 80% of arsenic accumulation in cytoplasm. The lack of arsC gene and arsenate reductase activity indicated that Bacillus sp. strain DJ-1 may lack classical ars operon and detoxification may be mediated through some novel mechanism. The arsenite binding protein was purified by affinity chromatography and characterized as DNA protection during starvation (DPS) protein by electrospray ionization mass spectrometry. The induction of DPS showed the adaptation of bacteria in arsenic stress condition and/or in detoxification mechanism, relies on its ability to bind with arsenic. These results indicate the hypertolerance with higher intracellular accumulation of arsenic by Bacillus sp. strain DJ-1, which could be mediated by DPS protein thus signifying this organism is a potential candidate for the removal of arsenic from industrial wastewater, which needs further study.

  3. Probiotic Bacillus cereus Strains, a Potential Risk for Public Health in China.

    Science.gov (United States)

    Zhu, Kui; Hölzel, Christina S; Cui, Yifang; Mayer, Ricarda; Wang, Yang; Dietrich, Richard; Didier, Andrea; Bassitta, Rupert; Märtlbauer, Erwin; Ding, Shuangyang

    2016-01-01

    Bacillus cereus is an important cause of foodborne infectious disease and food poisoning. However, B. cereus has also been used as a probiotic in human medicine and livestock production, with low standards of safety assessment. In this study, we evaluated the safety of 15 commercial probiotic B. cereus preparations from China in terms of mislabeling, toxin production, and transferable antimicrobial resistance. Most preparations were incorrectly labeled, as they contained additional bacterial species; one product did not contain viable B. cereus at all. In total, 18 B. cereus group strains-specifically B. cereus and Bacillus thuringiensis-were isolated. Enterotoxin genes nhe, hbl, and cytK1, as well as the ces-gene were assessed by PCR. Enterotoxin production and cytotoxicity were confirmed by ELISA and cell culture assays, respectively. All isolated B. cereus group strains produced the enterotoxin Nhe; 15 strains additionally produced Hbl. Antimicrobial resistance was assessed by microdilution; resistance genes were detected by PCR and further characterized by sequencing, transformation and conjugation assays. Nearly half of the strains harbored the antimicrobial resistance gene tet(45). In one strain, tet(45) was situated on a mobile genetic element-encoding a site-specific recombination mechanism-and was transferable to Staphylococcus aureus and Bacillus subtilis by electro-transformation. In view of the wide and uncontrolled use of these products, stricter regulations for safety assessment, including determination of virulence factors and transferable antimicrobial resistance genes, are urgently needed.

  4. Degradation characteristics of two Bacillus strains on the Microcystis aeruginosa

    Institute of Scientific and Technical Information of China (English)

    PEI Hai-yan; HU Wen-rong; QU Yin-bo; MU Rui-min; LI Xiao-cai

    2005-01-01

    The degradation kinetics of strains P05 and P07 and the degradation effects of mixed strain on Microcystis aeruginosa were studied. The results showed that: ( 1 ) The degradation processes of strains P05 and P07 on Microcystis aeruginosa accorded with the first-order reaction model when the range of Chl- a concentration was from 0 to 1500 μg/L. (2) The initial bacterium densities had a strong influence on the degradation velocity. The greater the initial bacterium density was, the faster the degradation was. The degradation velocity constants of P05 were 0.1913, 0.2175 and 0.3092 respectively, when bacterium densities were 4.8×105 , 4.8 × 106, 2.4 × 107 cells/ml. For strain P07, they were 0.1509, 0.1647 and 0.2708. The degradation velocity constant of strain P05 was higher than that of P07 when the bacterium density was under 4.8 × 105 cells/ml, but the constant increasing of P07 was quicker than that of P05. (3) The degradation effects of P05 and P07 strains did not antagonize. When the concentration of Chl-a was high, the degradation effects of mixed strain excelled that of any single strains. But with the decrease of the Chl-a concentration, this advantage was not clear. When the concentration was less than 180 μg/L, the degradation effects of mixed were consistent with that of strain P07.

  5. Complete Genome Sequence of Bacillus thuringiensis subsp. chinensis Strain CT-43▿

    Science.gov (United States)

    He, Jin; Wang, Jieping; Yin, Wen; Shao, Xiaohu; Zheng, Huajun; Li, Mingshun; Zhao, Youwen; Sun, Ming; Wang, Shengyue; Yu, Ziniu

    2011-01-01

    Bacillus thuringiensis has been widely used as an agricultural biopesticide for a long time. As a producing strain, B. thuringiensis subsp. chinensis strain CT-43 is highly toxic to lepidopterous and dipterous insects. It can form various parasporal crystals consisting of Cry1Aa3, Cry1Ba1, Cry1Ia14, Cry2Aa9, and Cry2Ab1. During fermentation, it simultaneously generates vegetative insecticidal protein Vip3Aa10 and the insecticidal nucleotide analogue thuringiensin. Here, we report the finished, annotated genome sequence of B. thuringiensis strain CT-43. PMID:21551307

  6. Complete genome sequence of Bacillus thuringiensis subsp. chinensis strain CT-43.

    Science.gov (United States)

    He, Jin; Wang, Jieping; Yin, Wen; Shao, Xiaohu; Zheng, Huajun; Li, Mingshun; Zhao, Youwen; Sun, Ming; Wang, Shengyue; Yu, Ziniu

    2011-07-01

    Bacillus thuringiensis has been widely used as an agricultural biopesticide for a long time. As a producing strain, B. thuringiensis subsp. chinensis strain CT-43 is highly toxic to lepidopterous and dipterous insects. It can form various parasporal crystals consisting of Cry1Aa3, Cry1Ba1, Cry1Ia14, Cry2Aa9, and Cry2Ab1. During fermentation, it simultaneously generates vegetative insecticidal protein Vip3Aa10 and the insecticidal nucleotide analogue thuringiensin. Here, we report the finished, annotated genome sequence of B. thuringiensis strain CT-43.

  7. Complete genome sequence of Bacillus thuringiensis serovar galleriae strain HD-29, a typical strain of commercial biopesticide.

    Science.gov (United States)

    Zhu, Lei; Tian, Long-Jun; Zheng, Jinshui; Gao, Qiu-Ling; Wang, Yue-Ying; Peng, Dong-Hai; Ruan, Li-Fang; Sun, Ming

    2015-02-10

    Bacillus thuringiensis serovar galleriae is highly toxic to Lepidoptera insect pests, and has been widely used as Bt biopesticide in many countries. Here we reported the complete genome of strain HD-29, a standard serotype strain in galleriae serovariety. More than previous work reported, it harbors ten plasmids, and three large ones carry eight insecticidal protein genes (cry1Aa, cry1Ac, cry1Ca, cry1Da, cry1Ia, cry2Ab, cry9Ea and vip3Aa) and an intact zwittermicin A biosynthetic gene cluster.

  8. A novel multiplex PCR discriminates Bacillus anthracis and its genetically related strains from other Bacillus cereus group species.

    Directory of Open Access Journals (Sweden)

    Hirohito Ogawa

    Full Text Available Anthrax is an important zoonotic disease worldwide that is caused by Bacillus anthracis, a spore-forming pathogenic bacterium. A rapid and sensitive method to detect B. anthracis is important for anthrax risk management and control in animal cases to address public health issues. However, it has recently become difficult to identify B. anthracis by using previously reported molecular-based methods because of the emergence of B. cereus, which causes severe extra-intestinal infection, as well as the human pathogenic B. thuringiensis, both of which are genetically related to B. anthracis. The close genetic relation of chromosomal backgrounds has led to complexity of molecular-based diagnosis. In this study, we established a B. anthracis multiplex PCR that can screen for the presence of B. anthracis virulent plasmids and differentiate B. anthracis and its genetically related strains from other B. cereus group species. Six sets of primers targeting a chromosome of B. anthracis and B. anthracis-like strains, two virulent plasmids, pXO1 and pXO2, a bacterial gene, 16S rRNA gene, and a mammalian gene, actin-beta gene, were designed. The multiplex PCR detected approximately 3.0 CFU of B. anthracis DNA per PCR reaction and was sensitive to B. anthracis. The internal control primers also detected all bacterial and mammalian DNAs examined, indicating the practical applicability of this assay as it enables monitoring of appropriate amplification. The assay was also applied for detection of clinical strains genetically related to B. anthracis, which were B. cereus strains isolated from outbreaks of hospital infections in Japan, and field strains isolated in Zambia, and the assay differentiated B. anthracis and its genetically related strains from other B. cereus group strains. Taken together, the results indicate that the newly developed multiplex PCR is a sensitive and practical method for detecting B. anthracis.

  9. A novel multiplex PCR discriminates Bacillus anthracis and its genetically related strains from other Bacillus cereus group species.

    Science.gov (United States)

    Ogawa, Hirohito; Fujikura, Daisuke; Ohnuma, Miyuki; Ohnishi, Naomi; Hang'ombe, Bernard M; Mimuro, Hitomi; Ezaki, Takayuki; Mweene, Aaron S; Higashi, Hideaki

    2015-01-01

    Anthrax is an important zoonotic disease worldwide that is caused by Bacillus anthracis, a spore-forming pathogenic bacterium. A rapid and sensitive method to detect B. anthracis is important for anthrax risk management and control in animal cases to address public health issues. However, it has recently become difficult to identify B. anthracis by using previously reported molecular-based methods because of the emergence of B. cereus, which causes severe extra-intestinal infection, as well as the human pathogenic B. thuringiensis, both of which are genetically related to B. anthracis. The close genetic relation of chromosomal backgrounds has led to complexity of molecular-based diagnosis. In this study, we established a B. anthracis multiplex PCR that can screen for the presence of B. anthracis virulent plasmids and differentiate B. anthracis and its genetically related strains from other B. cereus group species. Six sets of primers targeting a chromosome of B. anthracis and B. anthracis-like strains, two virulent plasmids, pXO1 and pXO2, a bacterial gene, 16S rRNA gene, and a mammalian gene, actin-beta gene, were designed. The multiplex PCR detected approximately 3.0 CFU of B. anthracis DNA per PCR reaction and was sensitive to B. anthracis. The internal control primers also detected all bacterial and mammalian DNAs examined, indicating the practical applicability of this assay as it enables monitoring of appropriate amplification. The assay was also applied for detection of clinical strains genetically related to B. anthracis, which were B. cereus strains isolated from outbreaks of hospital infections in Japan, and field strains isolated in Zambia, and the assay differentiated B. anthracis and its genetically related strains from other B. cereus group strains. Taken together, the results indicate that the newly developed multiplex PCR is a sensitive and practical method for detecting B. anthracis.

  10. A case of intoxication due to a highly cytotoxic Bacillus cereus strain isolated from cooked chicken.

    Science.gov (United States)

    López, Ana C; Minnaard, Jessica; Pérez, Pablo F; Alippi, Adriana M

    2015-04-01

    Outbreaks of Bacillus cereus infection/intoxication are not commonly reported because symptoms are often mild, and the disease is self-limiting. However, hypervirulent strains increase health risks. We report a case, which occurred in Argentina, of severe food poisoning illness on a healthy adult woman associated to B. cereus strain MVL2011. The studied strain was highly cytotoxic, showed high ability to detach Caco-2 cells and was positive for the hblA, hblB, and hblC genes of the hbl complex, bceT, entS and ces. As it is considered that B. cereus emetic cluster evolved from a panmictic population of diarrheal strains, B. cereus MVL2011 could constitute an intermediate strain between diarrheal and emetic strains.

  11. Diversity of mercury resistance determinants among Bacillus strains isolated from sediment of Minamata Bay.

    Science.gov (United States)

    Narita, Masaru; Chiba, Kazuyuki; Nishizawa, Hiroshi; Ishii, Hidenori; Huang, Chieh-Chen; Kawabata, Zen'ichiro; Silver, Simon; Endo, Ginro

    2003-06-06

    Thirty mercury-resistant (Hg R) Bacillus strains were isolated from mercury-polluted sediment of Minamata Bay, Japan. Mercury resistance phenotypes were classified into broad-spectrum (resistant to inorganic Hg(2+) and organomercurials) and narrow-spectrum (resistant to inorganic Hg(2+) and sensitive to organomercurials) groups. Polymerase chain reaction (PCR) product sizes and the restriction nuclease site maps of mer operon regions from all broad-spectrum Hg R Bacillus were identical to that of Bacillus megaterium MB1. On the other hand, the PCR products of the targeted merP (extracellular mercury-binding protein gene) and merA (intracellular mercury reductase protein gene) regions from the narrow-spectrum Hg R Bacillus were generally smaller than those of the B. megaterium MB1 mer determinant. Diversity of gene structure configurations was also observed by restriction fragment length polymorphism (RFLP) profiles of the merA PCR products from the narrow-spectrum Hg R Bacillus. The genetic diversity of narrow-spectrum mer operons was greater than that of broad-spectrum ones.

  12. Complete Genome Sequence of a Potential Novel Bacillus sp. Strain, FJAT-18017, Isolated from a Potato Field

    Science.gov (United States)

    Liu, Guo-Hong; Wang, Jie-Ping; Che, Jian-Mei; Chen, Qian-Qian

    2017-01-01

    ABSTRACT Bacillus sp. strain FJAT-18017 was isolated from a potato field in Xinjiang, China. This paper is the first report, to our knowledge, to demonstrate the fully sequenced and completely annotated genome of Bacillus sp. FJAT-18017. The genome size is 5,265,521 bp. The average G+C content was 42.42%. PMID:28104649

  13. Identification of Bacillus strains by MALDI TOF MS using geometric approach

    Science.gov (United States)

    Starostin, Konstantin V.; Demidov, Evgeny A.; Bryanskaya, Alla V.; Efimov, Vadim M.; Rozanov, Alexey S.; Peltek, Sergey E.

    2015-01-01

    Microorganism identification by MALDI TOF mass-spectrometry is based on the comparison of the mass spectrum of the studied organism with those of reference strains. It is a rapid and reliable method. However, commercial databases and programs are mostly designed for identification of clinically important strains and can be used only for particular mass spectrometer models. The need for open platforms and reference databases is obvious. In this study we describe a geometric approach for microorganism identification by mass spectra and demonstrate its capabilities by analyzing 24 strains belonging to the Bacillus pumilus group. This method is based on representing mass spectra as points on a multidimensional space, which allows us to use geometric distances to compare the spectra. Delimitation of microorganisms performed by geometric approach correlates well with the results of molecular phylogenetic analysis and clustering using Biotyper 3.1. All three methods used allowed us to reliably divide the strains into two groups corresponding to closely related species, Bacillus pumilus and Bacillus altitudinis. The method developed by us will be implemented in a Web interface designed for using open reference databases for microorganism identification. The data is available at http://www.bionet.nsc.ru/mbl/database/database.html. PMID:26592761

  14. Identification of Bacillus strains by MALDI TOF MS using geometric approach

    Science.gov (United States)

    Starostin, Konstantin V.; Demidov, Evgeny A.; Bryanskaya, Alla V.; Efimov, Vadim M.; Rozanov, Alexey S.; Peltek, Sergey E.

    2015-11-01

    Microorganism identification by MALDI TOF mass-spectrometry is based on the comparison of the mass spectrum of the studied organism with those of reference strains. It is a rapid and reliable method. However, commercial databases and programs are mostly designed for identification of clinically important strains and can be used only for particular mass spectrometer models. The need for open platforms and reference databases is obvious. In this study we describe a geometric approach for microorganism identification by mass spectra and demonstrate its capabilities by analyzing 24 strains belonging to the Bacillus pumilus group. This method is based on representing mass spectra as points on a multidimensional space, which allows us to use geometric distances to compare the spectra. Delimitation of microorganisms performed by geometric approach correlates well with the results of molecular phylogenetic analysis and clustering using Biotyper 3.1. All three methods used allowed us to reliably divide the strains into two groups corresponding to closely related species, Bacillus pumilus and Bacillus altitudinis. The method developed by us will be implemented in a Web interface designed for using open reference databases for microorganism identification. The data is available at http://www.bionet.nsc.ru/mbl/database/database.html.

  15. Mechanisms of microbial oil recovery by Clostridium acetobutylicum and Bacillus strain JF-2

    Energy Technology Data Exchange (ETDEWEB)

    Marsh, T.L.; Zhang, X.; Knapp, R.M.; McInerney, M.J.; Sharma, P.K.; Jackson, B.E.

    1995-12-31

    Core displacement experiments at elevated pressures were conducted to determine whether microbial processes are effective under conditions that simulate those found in an actual oil reservoir. The in-situ growth of Clostridium acetobutylicum and Bacillus strain JF-2 resulted in the recovery of residual oil. About 21 and 23% of the residual oil was recovered by C. acetobutylicum and Bacillus strain JF-2, respectively. Flooding cores with cell-free culture fluids of C. acetobutylicum with and without the addition of 50 mM acetone and 100 mM butanol did not result in the recovery of residual oil. Mathematical simulations showed that the amount of gas produced by the clostridial fermentation was not showed that the amount of gas produced by the clostridial fermentation was not sufficient to recover residual oil. Oil recovery by Bacillus strain JF-2 was highly correlated to surfactant production. A biosurfactant-deficient mutant of strain JF-2 was not capable of recovering residual oil. These data show that surfactant production is an important mechanism for microbially enhanced oil recovery. The mechanism for oil recovery by C. acetobutylicum is not understood at this time, but the production of acids, solvents, or gases alone cannot explain the observed increases in oil recovery by this organism.

  16. Selection of a Bacillus pumilus strain highly active against Ceratitis capitata (Wiedemann) larvae.

    Science.gov (United States)

    Molina, C Alfonso; Caña-Roca, Juan F; Osuna, Antonio; Vilchez, Susana

    2010-03-01

    Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), the Mediterranean fruit fly (medfly), is one of the most important fruit pests worldwide. The medfly is a polyphagous species that causes losses in many crops, which leads to huge economic losses. Entomopathogenic bacteria belonging to the genus Bacillus have been proven to be safe, environmentally friendly, and cost-effective tools to control pest populations. As no control method for C. capitata based on these bacteria has been developed, isolation of novel strains is needed. Here, we report the isolation of 115 bacterial strains and the results of toxicity screening with adults and larvae of C. capitata. As a result of this analysis, we obtained a novel Bacillus pumilus strain, strain 15.1, that is highly toxic to C. capitata larvae. The toxicity of this strain for C. capitata was related to the sporulation process and was observed only when cultures were incubated at low temperatures before they were used in a bioassay. The mortality rate for C. capitata larvae ranged from 68 to 94% depending on the conditions under which the culture was kept before the bioassay. Toxicity was proven to be a special characteristic of the newly isolated strain, since other B. pumilus strains did not have a toxic effect on C. capitata larvae. The results of the present study suggest that B. pumilus 15.1 could be considered a strong candidate for developing strategies for biological control of C. capitata.

  17. Construction of an environmental safe Bacillus thuringiensis engineered strain against Coleoptera.

    Science.gov (United States)

    Yu, Yajun; Yuan, Yihui; Gao, Meiying

    2016-05-01

    Cloning of new toxic genes from Bacillus thuringiensis (Bt) and construction of Bt engineered strains are two key strategies for bio-control of coleopteran pests in agriculture and forestry. In this study, we cloned a new cry3Aa-type gene, cry3Aa8, from wild Bt strain YC-03 against coleopteran, and constructed a Bt engineered strain, ACE-38, containing insecticidal protein-encoding gene cry3Aa8. The engineered strain, with almost four times of Cry3Aa yield compared with strain YC-03, was an antibiotic marker-free strain. Though no selective pressure was presented in the medium, cry3Aa8 in the engineered strain ACE-38 remained stable. The yield of Cry3Aa by strain ACE-38 reached 2.09 mg/ml in the optimized fermentation medium. The activity of strain ACE-38 against Plagiodera versicolora was tested, and the LC50 of ACE-38 cultures in the optimized fermentation medium was 1.13 μl/ml. Strain ACE-38 is a non-antibiotic Bt engineered strain with high Chrysomelidae toxicity and exhibits good fermentation property. The modified indigenous site-specific recombination system constructed in this study might be useful for the construction of Bt engineered strains containing genes that cannot be expressed in the indigenous site-specific recombination system using plasmid pBMB1205R.

  18. The structure of the transposable genetic element ISBsu2 from the cryptic plasmid p1516 of a soil Bacillus subtilis strain and the presence of homologues of this element in the chromosomes of various Bacillus subtilis strains

    NARCIS (Netherlands)

    Holsappel, S; Gagarina, EY; Poluektova, EU; Nezametdinova, VZ; Gel'fand, MS; Prozorov, AA; Bron, S

    2003-01-01

    A cryptic plasmid from a soil strain of Bacillus subtilis was found to contain a sequence having features of an IS element. Homologous sequences were also found in the chromosome of this strain and in the chromosomes of some other B. subtilis strains.

  19. Spores from mesophilic Bacillus cereus strains germinate better and grow faster in simulated gastro-intestinal conditions than spores from psychrotrophic strains.

    NARCIS (Netherlands)

    Wijnands, L M; Dufrenne, J B; Zwietering, M H; Leusden, F M van

    2006-01-01

    The species Bacillus cereus, known for its ability to cause food borne disease, consists of a large variety of strains. An important property for discrimination of strains is their growth temperature range. Psychrotrophic strains can grow well at refrigerator temperatures but grow at 37 degrees C wi

  20. Spores from mesophilic Bacillus cereus strains germinate better and grow faster in simulated gastro-intestinal conditions than spores from psychrotrophic strains

    NARCIS (Netherlands)

    Wijnands, L.M.; Dufrenne, J.B.; Zwietering, M.H.; Leusden, van F.M.

    2006-01-01

    The species Bacillus cereus, known for its ability to cause food borne disease, consists of a large variety of strains. An important property for discrimination of strains is their growth temperature range. Psychrotrophic strains can grow well at refrigerator temperatures but grow at 37 °C with diff

  1. Toxigenic genes, spoilage potential, and antimicrobial resistance of Bacillus cereus group strains from ice cream.

    Science.gov (United States)

    Arslan, Seza; Eyi, Ayla; Küçüksarı, Rümeysa

    2014-02-01

    Bacillus spp. can be recovered from almost every environment. It is also found readily in foods, where it may cause food spoilage and/or food poisoning due to its toxigenic and pathogenic nature, and extracellular enzymes. In this study, 29 Bacillus cereus group strains from ice cream were examined for the presence of following virulence genes hblC, nheA, cytK and ces genes, and tested for a range of the extracellular enzymes, and antimicrobial susceptibility. The strains were found to produce extracellular enzymes: proteolytic and lipolytic activity, gelatin hydrolysis and lecithinase production (100%), DNase production (93.1%) and amylase activity (93.1%). Of 29 strains examined, 24 (82.8%) showed hemolytic activity on blood agar. Beta-lactamase enzyme was only produced by 20.7% of B. cereus group. Among 29 B. cereus group from ice cream, nheA was the most common virulence gene detected in 44.8% of the strains, followed by hblC gene with 17.2%. Four (13.8%) of the 29 strains were positive for both hblC gene and nheA gene. Contrarily, cytK and ces genes were not detected in any of the strains. Antimicrobial susceptibility of ice cream isolates was tested to 14 different antimicrobial agents using the disc diffusion method. We detected resistance to penicillin and ampicillin with the same rate of 89.7%. Thirty-one percent of the strains were multiresistant to three or more antibiotics. This study emphasizes that the presence of natural isolates of Bacillus spp. harboring one or more enterotoxin genes, producing extracellular enzymes which may cause spoilage and acquiring antibiotic resistance might hold crucial importance in the food safety and quality.

  2. 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.

  3. Survival and conjugal transfer between Bacillus thuringiensis strains in aquatic environment

    OpenAIRE

    Furlaneto Luciana; Saridakis Halha Ostrensky; Arantes Olívia Márcia Nagy

    2000-01-01

    Field and laboratory studies were conducted to assess the survival of cells and spores and plasmid transfer between Bacillus thuringienis strains in aquatic environment. Results indicated that cells and spores of B. thuringiensis can survive for 10 days in water, without altering their number. The sporulation process began after 12-15 hours of inoculation of water. B. thuringiensis was able to transfer conjugative plasmids in the aquatic environment.

  4. Prediction of insecticidal activity of Bacillus thuringiensis strains by polymerase chain reaction product profiles.

    OpenAIRE

    Carozzi, N B; Kramer, V C; Warren, G W; Evola, S; Koziel, M G

    1991-01-01

    A rapid analysis of Bacillus thuringiensis strains predictive of insecticidal activity was established by using polymerase chain reaction (PCR) technology. Primers specific to regions of high homology within genes encoding three major classes of B. thuringiensis crystal proteins were used to generate a PCR product profile characteristic of each insecticidal class. Predictions of insecticidal activity were made on the basis of the electrophoretic patterns of the PCR products. Included in the s...

  5. 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.

  6. Study on the Flocculability of Metal Ions by Bacillus Mucilaginosus GY03 Strain

    Institute of Scientific and Technical Information of China (English)

    连宾; 陈烨; 袁生; 朱立军; 刘丛强

    2004-01-01

    This study deals with the flocculability of two types of metal ions by Bacillus mucilaginosus GY03 Strain, with an emphasis on the influence of pH conditions, volume and time of flocculants produced by GY03 Strain on the adsorption of metal irons such as Pb+2 and Mn+2 and the capabilities of flocculants to adsorb metal ions of different concentrations. The results showed that microbial flocculants produced by Bacillus mucilaginosus GY03 Strain are highly capable of flocculating metal ions, but show different effectiveness with respect to the adsorption of Pb+2 and Mn+2. In accordance with the experimental data and actual waste-water treatment conditions, the relevant regression equation of flocculation has been deduced, which has found some application in practice. The experimental results of this study demonstrated that microbial flocculants produced by Bacillus mucilaginosus can be used to treat metallic ion-containing waste water. In practical application the volume of microbial flocculants required and flocculation conditions should be taken into comprehensive consideration in accordance with the properties of metal ions, the composition of anions and the solubility of other metals, in combination with the cost and effectiveness of flocculants to be used. Flocculant used in this experiment has the advantages of being applied over a wide range of pH values, small flocculant volume, and rapid speed of flocculation. So this kind of flocculant is within excellent prospect of application.

  7. A Study on Effect of different culture media on amylase enzyme production by a native strain of Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    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.

  8. Bacillus flexus strain As-12, a new arsenic transformer bacterium isolated from contaminated water resources.

    Science.gov (United States)

    Jebeli, Mohammad Ahmadi; Maleki, Afshin; Amoozegar, Mohammad Ali; Kalantar, Enayatollah; Izanloo, Hassan; Gharibi, Fardin

    2017-02-01

    A total of 14 arsenic-resistant bacteria were isolated from an arsenic-contaminated travertine spring water in the central district of Qorveh county, Kurdistan Province, Iran. One of strains designated As-12 was selected for further investigation because of its ability to transform arsenic. The strain was identified by cultural, morphological and biochemical tests, and 16S rRNA gene sequencing. Finally, the growth characteristics of the isolate were investigated in a chemically defined medium which included varied ranges of environmental factors such as pH, temperature and salinity. Moreover, the resistance of this strain to some heavy metals was evaluated. The bacterium was a Gram-positive, endospore-forming with all other characteristics of the genus Bacillus. It revealed maximum similarity at the 16S rRNA gene level with Bacillus flexus. The optimum growth of the strain was observed at 38 °C, pH 9 and 2% salinity. This strain was resistant to heavy metals such as zinc, chromium, lead, nickel, copper, mercuric and cadmium at concentrations of 15 mM, 15.5 mM, 11.5 mM, 12 mM, 11 mM, 5.5 mM, and 1 mM, respectively. The isolated bacterium was able to reduce As (V) to As (III) (about 28%) and oxidize As (III) to As (V) (about 45%) after 48 h of incubation at 37 °C. In conclusion, Bacillus flexus strain As-12, was identified as an arsenic transformer, for the first time.

  9. Characterization of Cry34/Cry35 Binary Insecticidal Proteins from Diverse Bacillus thuringiensis Strain Collections

    OpenAIRE

    Schnepf, H. Ernest; Lee, Stacey; Dojillo, JoAnna; Burmeister, Paula; Fencil, Kristin; Morera, Lisa; Nygaard, Linda; Narva, Kenneth E.; Wolt, Jeff D.

    2005-01-01

    Bacillus thuringiensis crystal proteins of the Cry34 and Cry35 classes function as binary toxins showing activity on the western corn rootworm, Diabrotica virgifera virgifera LeConte. We surveyed 6,499 B. thuringiensis isolates by hybridization for sequences related to cry35A genes, identifying 78 strains. Proteins of the appropriate molecular mass (ca. 44 kDa) for Cry35 were observed in 42 of the strains. Full-length, or nearly full-length, sequences of 34 cry34 genes and 16 cry35 genes were...

  10. Complete Genome Sequence of Bacillus thuringiensis subsp. chinensis Strain CT-43▿

    OpenAIRE

    He, Jin; Wang, Jieping; Yin, Wen; Shao, Xiaohu; Zheng, Huajun; Li, Mingshun; Zhao, Youwen; Sun, Ming; Wang, Shengyue; Yu, Ziniu

    2011-01-01

    Bacillus thuringiensis has been widely used as an agricultural biopesticide for a long time. As a producing strain, B. thuringiensis subsp. chinensis strain CT-43 is highly toxic to lepidopterous and dipterous insects. It can form various parasporal crystals consisting of Cry1Aa3, Cry1Ba1, Cry1Ia14, Cry2Aa9, and Cry2Ab1. During fermentation, it simultaneously generates vegetative insecticidal protein Vip3Aa10 and the insecticidal nucleotide analogue thuringiensin. Here, we report the finished...

  11. Bacillus rubiinfantis sp. nov. strain mt2T, a new bacterial species isolated from human gut

    Directory of Open Access Journals (Sweden)

    M. Tidjiani Alou

    2015-11-01

    Full Text Available Bacillus rubiinfantis sp. nov. strain mt2T is the type strain of B. rubiinfantis sp. nov., isolated from the fecal flora of a child with kwashiorkor in Niger. It is Gram-positive facultative anaerobic rod belonging to the Bacillaceae family. We describe the features of this organism alongside the complete genome sequence and annotation. The 4 311 083 bp long genome (one chromosome but no plasmid contains 4028 protein-coding gene and 121 RNA genes including nine rRNA genes.

  12. Biocontrol of geosmin-producing Streptomyces spp. by two Bacillus strains from Chinese liquor.

    Science.gov (United States)

    Zhi, Yan; Wu, Qun; Du, Hai; Xu, Yan

    2016-08-16

    Streptomyces spp. producing geosmin have been regarded as the most frequent and serious microbial contamination causing earthy off-flavor in Chinese liquor. It is therefore necessary to control the Streptomyces community during liquor fermentation. Biological control, using the native microbiota present in liquor making, appears to be a better solution than chemical methods. The objective of this study was to isolate native microbiota antagonistic toward Streptomyces spp. and then to evaluate the possible action mode of the antagonists. Fourteen Bacillus strains isolated from different Daqu (the fermentation starter) showed antagonistic activity against Streptomyces sampsonii, which is one of the dominant geosmin producers. Bacillus subtilis 2-16 and Bacillus amyloliquefaciens 1-45 from Maotai Daqu significantly inhibited the growth of S. sampsonii by 57.8% and 84.3% respectively, and effectively prevented the geosmin production in the simulated fermentation experiments (inoculation ratio 1:1). To probe the biocontrol mode, the ability of strain 2-16 and 1-45 to produce antimicrobial metabolites and to reduce geosmin in the fermentation system was investigated. Antimicrobial substances were identified as lipopeptides by ultra-performance liquid chromatography tandem electrospray ionization/quadrupole-time-of-flight mass spectrometry (UPLC-ESI/Q-TOF MS) and in vitro antibiotic assay. In addition, strains 2-16 and 1-45 were able to remove 45% and 15% of the geosmin respectively in the simulated solid-state fermentation. This study highlighted the potential of biocontrol, and how the use of native Bacillus species in Daqu could provide an eco-friendly method to prevent growth of Streptomyces spp. and geosmin contamination in Chinese liquor fermentation.

  13. Antagonism of Bacillus spp. isolated from marine biofilms against terrestrial phytopathogenic fungi.

    Science.gov (United States)

    Ortega-Morales, B O; Ortega-Morales, F N; Lara-Reyna, J; De la Rosa-García, S C; Martínez-Hernández, A; Montero-M, Jorge

    2009-01-01

    We aimed at determining the antagonistic behavior of bacteria derived from marine biofilms against terrestrial phytopathogenic fungi. Some bacteria closely related to Bacillus mojavensis (three isolates) and Bacillus firmus (one isolate) displayed antagonistic activity against Colletotrichum gloeosporioides ATCC 42374, selected as first screen organism. The four isolates were further quantitatively tested against C. gloeosporioides, Colletotrichum fragariae, and Fusarium oxysporum on two culture media, potato dextrose agar (PDA) and a marine medium-based agar [yeast extract agar (YEA)] at different times of growth of the antagonists (early, co-inoculation with the pathogen and late). Overall antagonistic assays showed differential susceptibility among the pathogens as a function of the type of culture media and time of colonization (P Bacillus sp. MC3B-22 displayed a greater antagonistic effect than the commercial biocontrol strain Bacillus subtilis G03 (Kodiak). Further incubation studies and scanning electronic microscopy revealed that Bacillus sp. MC3B-22 was able to colonize, multiply, and inhibit C. gloeosporioides ATCC 42374 when tested in a mango leaf assay, showing its potential for fungal biocontrol. Additional studies are required to definitively identify the active isolates and to determine their mode of antifungal action, safety, and biocompatibility.

  14. Characteristics and antimicrobial activity of Bacillus subtilis strains isolated from soil.

    Science.gov (United States)

    Todorova, Sevdalina; Kozhuharova, Lubka

    2010-07-01

    Antagonistic Bacillus strains were isolated from soil and analyzed for the purpose of determining whether they could be used as natural biological agents. Primary in vitro screening for antagonism of the isolates was performed against five phytopathogenic mould fungi. Strains TS 01 and ZR 02 exhibited the most pronounced inhibitory effects. They were identified as Bacillus subtilis on the basis of their morphological, cultural and physiology-biochemical properties as well as their hierarchical cluster analysis conducted by means of computer program SPSS. The antimicrobial activity of the strains from cultural medium and sterile filtrate were determined in vitro against a great number of predominantly phytopathogenic fungi and bacteria. TS 01 and ZR 02 strains exhibited very broad and at the same time degree varying antibiotic spectra of activities against both Gram-positive and Gram-negative microorganisms. Many of them were tested against sensitivity to the antimicrobial action of B. subtilis for the very first time. B. subtilis TS 01 and ZR 02 showed highest antifungal activity (sterile zone in diameter over 37 mm) against Alternaria solani, Botrytis cinerea, Monilia linhartiana 869, Phytophthora cryptogea 759/1 and Rhizoctonia sp. The most sensitive bacterial species were found to be Pseudomonas syringae pv. tomato Ro and Xanthomonas campestris with sterile zones 48.0 and 50.0 mm in diameter, respectively. The latter draws a conclusion that the isolated and identified Bacillus subtilis strains are promising natural biocontrol agents and should be further studied and tested for control of numerous plant diseases.

  15. Genomic analysis of thermophilic Bacillus coagulans strains: efficient producers for platform bio-chemicals.

    Science.gov (United States)

    Su, Fei; Xu, Ping

    2014-01-29

    Microbial strains with high substrate efficiency and excellent environmental tolerance are urgently needed for the production of platform bio-chemicals. Bacillus coagulans has these merits; however, little genetic information is available about this species. Here, we determined the genome sequences of five B. coagulans strains, and used a comparative genomic approach to reconstruct the central carbon metabolism of this species to explain their fermentation features. A novel xylose isomerase in the xylose utilization pathway was identified in these strains. Based on a genome-wide positive selection scan, the selection pressure on amino acid metabolism may have played a significant role in the thermal adaptation. We also researched the immune systems of B. coagulans strains, which provide them with acquired resistance to phages and mobile genetic elements. Our genomic analysis provides comprehensive insights into the genetic characteristics of B. coagulans and paves the way for improving and extending the uses of this species.

  16. Use of long-range repetitive element polymorphism-PCR to differentiate Bacillus anthracis strains.

    Science.gov (United States)

    Brumlik, M J; Szymajda, U; Zakowska, D; Liang, X; Redkar, R J; Patra, G; Del Vecchio, V G

    2001-07-01

    The genome of Bacillus anthracis is extremely monomorphic, and thus individual strains have often proven to be recalcitrant to differentiation at the molecular level. Long-range repetitive element polymorphism-PCR (LR REP-PCR) was used to differentiate various B. anthracis strains. A single PCR primer derived from a repetitive DNA element was able to amplify variable segments of a bacterial genome as large as 10 kb. We were able to characterize five genetically distinct groups by examining 105 B. anthracis strains of diverse geographical origins. All B. anthracis strains produced fingerprints comprising seven to eight bands, referred to as "skeleton" bands, while one to three "diagnostic" bands differentiated between B. anthracis strains. LR REP-PCR fingerprints of B. anthracis strains showed very little in common with those of other closely related species such as B. cereus, B. thuringiensis, and B. mycoides, suggesting relative heterogeneity among the non-B. anthracis strains. Fingerprints from transitional non-B. anthracis strains, which possessed the B. anthracis chromosomal marker Ba813, scarcely resembled those observed for any of the five distinct B. anthracis groups that we have identified. The LR REP-PCR method described in this report provides a simple means of differentiating B. anthracis strains.

  17. Cyclic lipopeptide profile of three Bacillus subtilis strains; antagonists of Fusarium head blight.

    Science.gov (United States)

    Dunlap, Christopher A; Schisler, David A; Price, Neil P; Vaughn, Steven F

    2011-08-01

    The objective of the study was to identify the lipopetides associated with three Bacillus subtilis strains. The strains are antagonists of Gibberella zeae, and have been shown to be effective in reducing Fusarium head blight in wheat. The lipopeptide profile of three B. subtilis strains (AS43.3, AS43.4, and OH131.1) was determined using mass spectroscopy. Strains AS43.3 and AS43.4 produced the anti-fungal lipopeptides from the iturin and fengycin family during the stationary growth phase. All three strains produced the lipopeptide surfactin at different growth times. Strain OH131.1 only produced surfactin under these conditions. The antifungal activity of the culture supernatant and individual lipopeptides was determined by the inhibition of G. zeae. Cell-free supernatant from strains AS43.3 and AS43.4 demonstrated strong antibiosis of G. zeae, while strain OH131.1 had no antibiosis activity. These results suggest a different mechanism of antagonism for strain OH131.1, relative to AS43.3 and AS43.4.

  18. In Vitro Screening for Abiotic Stress Tolerance in Potent Biocontrol and Plant Growth Promoting Strains of Pseudomonas and Bacillus spp.

    Directory of Open Access Journals (Sweden)

    G. Praveen Kumar

    2014-01-01

    Full Text Available Plant growth promoting rhizobacteria (PGPR has been identified as a group of microbes that are used for plant growth enhancement and biocontrol for management of plant diseases. The inconsistency in performance of these bacteria from laboratory to field conditions is compounded due to the prevailing abiotic stresses in the field. Therefore, selection of bacterial strains with tolerance to abiotic stresses would benefit the end-user by successful establishment of the strain for showing desired effects. In this study we attempted to isolate and identify strains of Bacillus and Pseudomonas spp. with stress tolerance and proven ability to inhibit the growth of potential phytopathogenic fungi. Screening of bacterial strains for high temperature (50°C, salinity (7% NaCl, and drought (−1.2 MPa showed that stress tolerance was pronounced less in Pseudomonas isolates than in Bacillus strains. The reason behind this could be the formation of endospores by Bacillus isolates. Tolerance to drought was high in Pseudomonas strains than the other two stresses. Three strains, P8, P20 and P21 showed both salinity and temperature tolerance. P59 strain possessed promising antagonistic activity and drought tolerance. The magnitude of antagonism shown by Bacillus isolates was also higher when compared to Pseudomonas strains. To conclude, identification of microbial candidate strains with stress tolerance and other added characteristic features would help the end-user obtain the desired beneficial effects.

  19. In Vitro Screening for Abiotic Stress Tolerance in Potent Biocontrol and Plant Growth Promoting Strains of Pseudomonas and Bacillus spp.

    Science.gov (United States)

    Praveen Kumar, G; Mir Hassan Ahmed, S K; Desai, Suseelendra; Leo Daniel Amalraj, E; Rasul, Abdul

    2014-01-01

    Plant growth promoting rhizobacteria (PGPR) has been identified as a group of microbes that are used for plant growth enhancement and biocontrol for management of plant diseases. The inconsistency in performance of these bacteria from laboratory to field conditions is compounded due to the prevailing abiotic stresses in the field. Therefore, selection of bacterial strains with tolerance to abiotic stresses would benefit the end-user by successful establishment of the strain for showing desired effects. In this study we attempted to isolate and identify strains of Bacillus and Pseudomonas spp. with stress tolerance and proven ability to inhibit the growth of potential phytopathogenic fungi. Screening of bacterial strains for high temperature (50°C), salinity (7% NaCl), and drought (-1.2 MPa) showed that stress tolerance was pronounced less in Pseudomonas isolates than in Bacillus strains. The reason behind this could be the formation of endospores by Bacillus isolates. Tolerance to drought was high in Pseudomonas strains than the other two stresses. Three strains, P8, P20 and P21 showed both salinity and temperature tolerance. P59 strain possessed promising antagonistic activity and drought tolerance. The magnitude of antagonism shown by Bacillus isolates was also higher when compared to Pseudomonas strains. To conclude, identification of microbial candidate strains with stress tolerance and other added characteristic features would help the end-user obtain the desired beneficial effects.

  20. Genomic signatures of strain selection and enhancement in Bacillus atrophaeus var. globigii, a historical biowarfare simulant.

    Directory of Open Access Journals (Sweden)

    Henry S Gibbons

    Full Text Available BACKGROUND: Despite the decades-long use of Bacillus atrophaeus var. globigii (BG as a simulant for biological warfare (BW agents, knowledge of its genome composition is limited. Furthermore, the ability to differentiate signatures of deliberate adaptation and selection from natural variation is lacking for most bacterial agents. We characterized a lineage of BGwith a long history of use as a simulant for BW operations, focusing on classical bacteriological markers, metabolic profiling and whole-genome shotgun sequencing (WGS. RESULTS: Archival strains and two "present day" type strains were compared to simulant strains on different laboratory media. Several of the samples produced multiple colony morphotypes that differed from that of an archival isolate. To trace the microevolutionary history of these isolates, we obtained WGS data for several archival and present-day strains and morphotypes. Bacillus-wide phylogenetic analysis identified B. subtilis as the nearest neighbor to B. atrophaeus. The genome of B. atrophaeus is, on average, 86% identical to B. subtilis on the nucleotide level. WGS of variants revealed that several strains were mixed but highly related populations and uncovered a progressive accumulation of mutations among the "military" isolates. Metabolic profiling and microscopic examination of bacterial cultures revealed enhanced growth of "military" isolates on lactate-containing media, and showed that the "military" strains exhibited a hypersporulating phenotype. CONCLUSIONS: Our analysis revealed the genomic and phenotypic signatures of strain adaptation and deliberate selection for traits that were desirable in a simulant organism. Together, these results demonstrate the power of whole-genome and modern systems-level approaches to characterize microbial lineages to develop and validate forensic markers for strain discrimination and reveal signatures of deliberate adaptation.

  1. Isolation and characterization of atrazine mineralizing Bacillus subtilis strain HB-6.

    Directory of Open Access Journals (Sweden)

    Jinhua Wang

    Full Text Available Atrazine is a widely used herbicide with great environmental concern due to its high potential to contaminate soil and waters. An atrazine-degrading bacterial strain HB-6 was isolated from industrial wastewater and the 16S rRNA gene sequencing identified HB-6 as a Bacillus subtilis. PCR assays indicated that HB-6 contained atrazine-degrading genes trzN, atzB and atzC. The strain HB-6 was capable of utilizing atrazine and cyanuric acid as a sole nitrogen source for growth and even cleaved the s-triazine ring and mineralized atrazine. The strain demonstrated a very high efficiency of atrazine biodegradation with a broad optimum pH and temperature ranges and could be enhanced by cooperating with other bacteria, suggesting its huge potential for remediation of atrazine-contaminated sites. To our knowledge, there are few Bacillus subtilis strains reported that can mineralize atrazine, therefore, the present work might provide some new insights on atrazine remediation.

  2. Isolation and characterization of atrazine mineralizing Bacillus subtilis strain HB-6.

    Science.gov (United States)

    Wang, Jinhua; Zhu, Lusheng; Wang, Qi; Wang, Jun; Xie, Hui

    2014-01-01

    Atrazine is a widely used herbicide with great environmental concern due to its high potential to contaminate soil and waters. An atrazine-degrading bacterial strain HB-6 was isolated from industrial wastewater and the 16S rRNA gene sequencing identified HB-6 as a Bacillus subtilis. PCR assays indicated that HB-6 contained atrazine-degrading genes trzN, atzB and atzC. The strain HB-6 was capable of utilizing atrazine and cyanuric acid as a sole nitrogen source for growth and even cleaved the s-triazine ring and mineralized atrazine. The strain demonstrated a very high efficiency of atrazine biodegradation with a broad optimum pH and temperature ranges and could be enhanced by cooperating with other bacteria, suggesting its huge potential for remediation of atrazine-contaminated sites. To our knowledge, there are few Bacillus subtilis strains reported that can mineralize atrazine, therefore, the present work might provide some new insights on atrazine remediation.

  3. Characterization of melanin produced by a wild-type strain of Bacillus cereus

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jianping; CAI Jun; DENG Yinyue; CHEN Yuehua; REN Gaixin

    2007-01-01

    Bacillus cereus 58 (Bc58)is a UV-resistant wild type strain that has an ability to produce a sorrel pigment induced by L-tyrosine.The Fourier-transform infrared (FT-IR)spectra and chemical tests of its pigment are similar to that of the standard melanin (Sigma).A bioassay shows that the LC50 of a Bacillus thuringiensis (Bt)formulation added with the melanin of Bc58 and exposed to UV for 5 h is 16.1 μg/ml,which is similar to that of the Bt formulation without UV treatment,however,it is almost double that of the Bt formulation exposed to UV without the melanin of Bc58.The result of SDS-PAGE indicates that the melanin of Bc58 can protect the insecticidal crystal proteins from degradation.This suggests that it is an excellent UV protective agent for the insecticidal crystal proteins of the Bt formulation.

  4. [Isolation and Identification of Petroleum Degradation Bacteria and Interspecific Interactions Among Four Bacillus Strains].

    Science.gov (United States)

    Wang, Jia-nan; Shi, Yan-yun; Zheng, Li-yan; Wang, Zhe; Cai, Zhang; Liu, Jie

    2015-06-01

    Six petroleum-degrading strains were isolated from oil-contaminated soil at Dagang oil field and oil sewage on Bohai offshore drilling platform in Tianjin using enrichment culture and isolation method. The physiological biochemical test together with 16S rDNA sequencing analysis indicated that they belonged to Bacillus (S1, S2, S3, S4), Pseudomonas (W1) and Ochrobactrum (W2), respectively. The strain S3 had the maximum degradation rate of alkane (41.3%) and aromatic hydrocarbon (30.9%) among all isolated strains showing the better degradation efficiency by endogenous bacteria when compared to that by the exogenous bacteria. The four Bacillus strains were used to construct microbiome, thereafter subjected to petroleum degradation efficiency test and analyzed. The results showed that microbiome F3 consisting of S1 and S4 had the maximum degradation rates of alkane (50.5%) and aromatic hydrocarbon (54.0%), which were 69.9% and 156.1% higher than those by single bacterium, respectively. Furthermore, they were 22.1% and 74.6% respectively higher than those by the most optimal degradation bacterium S3. Microbiome F4 consisting of S2 and S3 had the minimum degradation rates of alkane (18.5%) and aromatic hydrocarbon (18.9%) which were 55.3% and 39.0% lower than the degradation rates of single bacterium, respectively. The results also demonstrated that there were both microbial synergy promotion and antagonism inhibition among bacteria of the same genus in the petroleum degradation period. Bacteria with close affinity in Bacillus genus displayed mainly promoted petroleum degradation effect.

  5. An Ultra-Violet Tolerant Wild-Type Strain of Melanin-Producing Bacillus thuringiensis

    Science.gov (United States)

    Sansinenea, Estibaliz; Salazar, Francisco; Ramirez, Melanie; Ortiz, Aurelio

    2015-01-01

    Background: Bacillus thuringiensis is the most successful biological control agent used in agriculture, forestry and mosquito control. However, the insecticidal activity of the B. thuringiensis formulation is not very stable and rapidly loses its biological activity under field conditions, due to the ultraviolet radiation in sunlight. Melanin is known to absorb radiation therefore photo protection of B. thuringiensis based on melanin has been extensively studied. Objectives: The aim of this study was to find a wild type strain of naturally melanin-producing B. thuringiensis to avoid any mutation or manipulation that can affect the Cry protein content. Materials and Methods: Bacillus thuringiensis strains were isolated from soils of different States of Mexico and pigment extraction was followed by lowering the pH to 2 using 1N HCl. Pigment was characterized by some chemical tests based on its solubility, bleaching by H2O2 and flocculation with FeCl3, and using an Infrared (IR) spectrum. Ultraviolet (UV) irradiation experiment was performed to probe the melanin efficacy. Results: ELI52 strain of B. thuringiensis was confirmed to naturally produce melanin. The Cry protein analysis suggested that ELI52 is probably a B. thuringiensis subsp. israelensis strain with toxic activity against the Diptera order of insects. Ultra Violet protection efficacy of melanin was probed counting total viable colonies after UV radiation and comparing the results with the non-producing melanin strain L-DOPA (L-3, 4-dihydroxyphenylalanine) was also detected in the culture. ELI52 strain showed an antagonistic effect over some common bacteria from the environment. Conclusions: ELI52 wild-type strain of B. thuringiensis is a good bio-insecticide that produces melanin with UV-resistance that is probably toxic against the Diptera order of insects and can inhibit the growth of other environmental bacteria. PMID:26421136

  6. Invertebrate pathogenicity and toxin-producing potential of strains of Bacillus thuringiensis endemic to Antarctica.

    Science.gov (United States)

    Prabhakar, A; Bishop, A H

    2011-06-01

    Several strains of Bacillus thuringiensis were previously isolated from soil in Antarctica and appeared to have physiological adaptations to this cold, nutrient-poor environment. In spite of this they could produce abnormally large, parasporal crystals under laboratory conditions. Here, they have been further characterised for toxin genes and invertebrate pathogenicity. All of the strains were positive in PCR assays for the cry1Aa and cry2 genes. This was confirmed by sequence analysis and the parasporal crystals of all strains contained polypeptides of about 130kDa. This potential for lepidopteran toxicity was borne out in bioassays of purified δ-endotoxins against larvae of Pieris brassicae: the LD(50) values of B2408 (288μg) were comparable to that of the reference strain, HD-12 (201μg). There was no activity against the nematode Caenorhabditis elegans in spite of the fact that all strains appeared to possess the cry6 gene. PCR screening for genes encoding other nematode-toxic classes of toxins (Cry5, 4 and 21) was negative. B. thuringiensis has never previously been shown to be toxic to Collembola (springtails) but the purified δ-endotoxins of one of the Antarctic strains showed some activity against Folsomia candida and Seira domestica (224μg and 238μg, respectively). It seems unlikely that the level of toxicity demonstrated against springtails would support a pathogenic life-style in nature. All of the strains were positive for genes encoding Bacillus cereus-type enterotoxins. In the absence of higher insects and mammals the ecological value of retaining the toxic capability demonstrated here is uncertain.

  7. Use of bacillus subtilis strains to inhibit postharvest pathogenic fungi; Attivita` antagonista di alcuni ceppi di bacillus subtilis nei confronti di funghi patogeni

    Energy Technology Data Exchange (ETDEWEB)

    Arras, G.; Gambella, F.; Demontis, S.; Petretto, A.

    1995-09-01

    An isolate (87) of the bacillus subtilis strains isolated from cold stored citrus fruit 13 proved to inhibit the growth in vitro of the penicillium italicum used in the experiment (from 50.6% to 92.2%) and to inhibit botrytis cinerea (from 65.3% to 95.9%). A further test, superimposing on plates containing PDA strains Nos. 13, 173, and 160, totally inhibited the fungi. Tested in vivo on artificially bruised oranges, they significantly inhibited two fungi.

  8. Effects of nitrogen and carbon sources on the production of inulinase from strain Bacillus sp. SG113

    Science.gov (United States)

    Gavrailov, Simeon; Ivanova, Viara

    2016-03-01

    The effects of the carbon and nitrogen substrates on the growth of Bacillus sp. SG113 strain were studied. The use of organic nitrogen sources (peptone, beef extract, yeast extract, casein) leads to rapid cellular growth and the best results for the Bacillus strain were obtained with casein hydrolysate. From the inorganic nitrogen sources studied, the (NH4) 2SO4 proved to be the best nitrogen source. Casein hydrolysate and (NH4) 2SO4 stimulated the invertase synthesis. In the presence of Jerusalem artichoke, onion and garlic extracts as carbon sources the strain synthesized from 6 to 10 times more inulinase.

  9. Screening of bacterial strains for pectinolytic activity: characterization of the polygalacturonase produced by Bacillus sp

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    Soares Márcia M.C.N.

    1999-01-01

    Full Text Available One hundred sixty eight bacterial strains, isolated from soil and samples of vegetable in decomposition, were screened for the use of citrus pectin as the sole carbon source. 102 were positive for pectinase depolymerization in assay plates as evidenced by clear hydrolization halos. Among them, 30% presented considerable pectinolytic activity. The cultivation of these strains by submerged and semi-solid fermentation for polygalacturonase production indicated that five strains of Bacillus sp produced high quantities of the enzyme. The physico-chemical characteristics, such as optimum pH of 6.0 - 7.0, optimum temperatures between 45oC and 55oC, stability at temperatures above 40oC and in neutral and alkaline pH, were determined.

  10. Development of an efficient electroporation method for rhizobacterial Bacillus mycoides strains.

    Science.gov (United States)

    Yi, Yanglei; Kuipers, Oscar P

    2017-02-01

    In order to develop a method for electroporation of environmental Bacillus mycoides strains, we optimized several conditions that affect the electroporation efficiency of this bacterium. By combining the optimized conditions, the electroporation efficiency of strain EC18 was improved to (1.3±0.6)×10(5)cfu/μg DNA, which is about 10(3)-fold increase in comparison with a previously reported value. The method was further validated on various B. mycoides strains, yielding reasonable transformation efficiencies. Furthermore, we confirmed that restriction/modification is the main barrier for electroporation of this bacterium. To the best of our knowledge, this is the first systematic investigation of various parameters of electroporation of B. mycoides. The electroporation method reported will allow for efficient genetic manipulation of this bacterium.

  11. The ability of the biological control agent Bacillus subtilis, strain BB, to colonise vegetable brassicas endophytically following seed inoculation

    NARCIS (Netherlands)

    Wulff, E.G.; Vuurde, van J.W.L.; Hockenhull, J.

    2003-01-01

    The ability of Bacillus subtilis, strain BB, to colonise cabbage seedlings endophytically was examined following seed inoculation. Strain BB was recovered from different plant parts including leaves (cotyledons), stem (hypocotyl) and roots. While high bacterial populations persisted in the roots and

  12. Comparative analysis of biofilm formation by Bacillus cereus reference strains and undomesticated food isolates and the effect of free iron

    NARCIS (Netherlands)

    Hayrapetyan, H.; Muller, L.K.; Tempelaars, M.H.; Abee, T.; Nierop Groot, M.N.

    2015-01-01

    Biofilm formation of Bacillus cereus reference strains ATCC 14579 and ATCC 10987 and 21 undomesticated food isolates was studied on polystyrene and stainless steel as contact surfaces. For all strains, the biofilm forming capacity was significantly enhanced when in contact with stainless steel (SS)

  13. Selection and Molecular Biological Identification of a Strain of Bacillus sp. Inhibiting the Growth of Saprolegnia ferax

    Institute of Scientific and Technical Information of China (English)

    Song; Zengfu; Fan; Bin; She; Linrong; Tang; Lei; Zhao; Shilin; Lv; Liqun; Yang; Xianle

    2014-01-01

    Based on the theory of biological control of Saprolegnia ferax,antagonism test of nine strains of Bacillus sp. to S. ferax JL was carried out. Bacillus sp.BA1 was screened to have significantly inhibitory effects on the growth of S. ferax JL( P < 0. 05). Then,the effects of Bacillus sp. BA1 on different sources of S. ferax were carried out. Results showed that BA1 also had significantly inhibitory effects on S. ferax 6#,10# and S2( P < 0. 05). Sequence of 16 S r DNA of BA1 was analyzed; and homologous alignment analysis showed that BA1 had more than 99% similarity with Bacillus cereus. Therefore,it could be concluded that strain BA1 was B. cereus,which significantly inhibited the growth of S. ferax and could be used as the biological control agent for S. ferax diseases in aquaculture.

  14. [Extracellular hydrolases of strain Bacillus sp. 739 and their involvement in the lysis of micromycete cell walls].

    Science.gov (United States)

    Aktuganov, G E; Galimzianova, N F; Melent'ev, A I; Kuz'mina, L Iu

    2007-01-01

    The mycolytic bacterial strain Bacillus sp. 739 produces extracellular enzymes which degrade in vitro the cell walls of a number of phytopathogenic and saprophytic fungi. When Bacillus sp. 739 was cultivated with Bipolaris sorokiniana, a cereal root-rot pathogen, the fungus degradation process correlated with the levels of the beta-1,3-glucanase and protease activity. The comparative characteristic of Bacillus sp. 739 enzymatic preparations showed that efficient hydrolysis of the fungus cell walls was the result of the action of the complex of enzymes produced by the strain when grown on chitin-containing media. Among the enzymes of this complex, chitinases and beta-1,3-glucanases hydrolyzed most actively the disintegrated cell walls of B. sorokiniana. However, only beta-1,3-glucanases were able to degrade the cell walls of native fungal mycelium in the absence of other hydrolases, which is indicative of their key role in the mycolytic activity of Bacillus sp. 739.

  15. Bioremediation potential of a newly isolate solvent tolerant strain Bacillus thermophilus PS11

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    PAYEL SARKAR

    2012-01-01

    Full Text Available The increased generation of solvent waste has been stated as one of the most critical environmental problems. Though microbial bioremediation has been widely used for waste treatment but their application in solvent waste treatment is limited since the solvents have toxic effects on the microbial cells. A solvent tolerant strain of Bacillus thermophilus PS11 was isolated from soil by cyclohexane enrichment. Transmission electron micrograph of PS11 showed convoluted cell membrane and accumulation of solvents in the cytoplasm, indicating the adaptation of the bacterial strain to the solvent after 48h of incubation. The strain was also capable of growing in presence of wide range of other hydrophobic solvents with log P-values below 3.5. The isolate could uptake 50 ng/ml of uranium in its initial 12h of growth, exhibiting both solvent tolerance and metal resistance property. This combination of solvent tolerance and metal resistance will make the isolated Bacillus thermophilus PS11 a potential tool for metal bioremediation in solvent rich wastewaters.

  16. Screening of Bacterial Strains for Polygalacturonase Activity: Its Production by Bacillus sphaericus (MTCC 7542

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    Ranveer Singh Jayani

    2010-01-01

    Full Text Available At present almost all the pectinolytic enzymes used for industrial applications are produced by fungi. There are a few reports of pectinase production by bacterial strains. Therefore, in the present study, seventy-four bacterial strains, isolated from soil and rotten vegetable samples, were screened for polygalacturonase production. The strain PG-31, which gave maximum activity, was identified as Bacillus sphaericus (MTCC 7542. Maximal quantities of polygalacturonase were produced when a 16-hours-old inoculum was used at 7.5% (v/v in production medium and incubated in shaking conditions (160 rpm for 72 hours. The optimal temperature and pH for bacterial growth and polygalacturonase production were found to be 30∘C and 6.8, respectively. Maximum enzyme production resulted when citrus pectin was used as the carbon source at a concentration of 1.25% (w/v, whereas other carbon sources led to a decrease (30%–70% in enzyme production. Casein hydrolysate and yeast extract used together as organic nitrogen source gave best results, and ammonium chloride was found to be the most suitable inorganic nitrogen source. The supplementation of media with 0.9% (w/v D-galacturonic acid led to a 23% increase in activity. Bacillus sphaericus, a bacterium isolated from soil, produced good amount of polygalacturonase activity at neutral pH; hence, it would be potentially useful to increase the yield of banana, grape, or apple juice.

  17. Investigation of the Antimicrobial Activity of Bacillus licheniformis Strains Isolated from Retail Powdered Infant Milk Formulae.

    Science.gov (United States)

    Alvarez-Ordóñez, Avelino; Begley, Máire; Clifford, Tanya; Deasy, Thérèse; Considine, Kiera; O'Connor, Paula; Ross, R Paul; Hill, Colin

    2014-03-01

    This study investigated the potential antimicrobial activity of ten Bacillus licheniformis strains isolated from retail infant milk formulae against a range of indicator (Lactococcus lactis, Lactobacillus bulgaricus and Listeria innocua) and clinically relevant (Listeria monocytogenes, Staphylococcus aureus, Streptococcus agalactiae, Salmonella Typhimurium and Escherichia coli) microorganisms. Deferred antagonism assays confirmed that all B. licheniformis isolates show antimicrobial activity against the Gram-positive target organisms. PCR and matrix-assisted laser desorption ionization time-of-flight mass spectrometry analyses indicated that four of the B. licheniformis isolates produce the bacteriocin lichenicidin. The remaining six isolates demonstrated a higher antimicrobial potency than lichenicidin-producing strains. Further analyses identified a peptide of ~1,422 Da as the most likely bioactive responsible for the antibacterial activity of these six isolates. N-terminal sequencing of the ~1,422 Da peptide from one strain identified it as ILPEITXIFHD. This peptide shows a high homology to the non-ribosomal peptides bacitracin and subpeptin, known to be produced by Bacillus spp. Subsequent PCR analyses demonstrated that the six B. licheniformis isolates may harbor the genetic machinery needed for the synthesis of a non-ribosomal peptide synthetase similar to those involved in production of subpeptin and bacitracin, which suggests that the ~1,422 Da peptide might be a variant of subpeptin and bacitracin.

  18. The Construction of the Probe for Amylase Ⅱ Gene Cloning from Bacillus halodurans Strain 38C1-1

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Primers and probes were established according to the sequences of the alpha-amylase genes of Bacillus. halodurans C-125, Thermus sp. IM6501, B. stearothermophilus ET-1, and B. acidopullulytics. Primers were designed and a 0.2 kb DNA fragment was amplified, the fragment was successfully used for the detection of the amylase Ⅱ gene in a 2 842 bp region from Bacillus halodurans strain 38C1-1.

  19. Bacillus cereus strain isolated from Demodex folliculorum in patients with topical steroid-induced rosaceiform facial dermatitis.

    Science.gov (United States)

    Tatu, Alin Laurentiu; Ionescu, Marius Anton; Clatici, Victor Gabriel; Cristea, Violeta Corina

    2016-01-01

    The aim of the study was to identify Bacillus species from the Demodex folliculorum of patients with topical steroidinduced facial rosaceiform dermatitis. Of the 75 patients examined, 20% had clinical spinulosis, while 18.66% had dermoscopic features of Demodex: follicular plugs and tails. Of the 17.33% positive patients identified upon microscopy for Demodex, samples for bacterial culture were plated on trypticase soy Colombia agar. Identification was performed by microorganisms grown method mass spectrometry. We identified a strain of Bacillus cereus.

  20. Draft genome sequence of Bacillus thuringiensis 147, a Brazilian strain with high insecticidal activity

    Science.gov (United States)

    Barbosa, Luiz Carlos Bertucci; Farias, Débora Lopes; Silva, Isabella de Moraes Guimarães; Melo, Fernando Lucas; Ribeiro, Bergmann Morais; Aguiar, Raimundo Wagner de Souza

    2015-01-01

    Bacillus thuringiensis is a ubiquitous Gram-positive and sporulating bacterium. Its crystals and secreted toxins are useful tools against larvae of diverse insect orders and, as a consequence, an alternative to recalcitrant chemical insecticides. We report here the draft genome sequence ofB. thuringiensis 147, a strain isolated from Brazil and with high insecticidal activity. The assembled genome contained 6,167,994 bp and was distributed in seven replicons (a chromosome and 6 plasmids). We identified 12 coding regions, located in two plasmids, which encode insecticidal proteins. PMID:26517667

  1. A Soluble Aggregated Thermophile Metalloaminopeptidase Produced by an Alcalophile Strain of Bacillus halodurans

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    S. Dabonné

    2011-01-01

    Full Text Available H4 strain isolated from Lake Bogoria was found to be Bacillus halodurans. The Bacteria produced an extracellular peptidase activity toward substrates Ile-pNA, Met-pNA and Val-pNA. It also hydrolyzed small peptides. A purification procedure including ion-exchange chromatography ion exchange DEAE and sizeexclusion chromatography followed by Sodium dodecyl sulphate-polyacrymalide gel electrophoresis (SDSPAGE revealed the aggregated form of the enzyme. The three substrates are hydrolyzed by a single catalytic site. The enzyme inactivated by bestatin, and 1,10-phenanthroline is a metalloaminopeptidase whose activity is maximal at pH 9.0 and 65ºC.

  2. Population transcriptomics of cactus host shifts in Drosophila mojavensis.

    Science.gov (United States)

    Matzkin, Luciano M

    2012-05-01

    In the presence of environmental change, natural selection can shape the transcriptome. Under a scenario of environmental change, genotypes that are better able to modulate gene expression to maximize fitness will tend to be favoured. Therefore, it is important to examine gene expression at the population level to distinguish random or neutral gene expression variation from the pattern produced by natural selection. This study investigates the natural variation in transcriptional response to a cactus host shift utilizing the mainland Sonora population of Drosophila mojavensis. Drosophila mojavensis is a cactophilic species composed of four cactus host populations endemic to the deserts of North America. Overall, the change in cactus host was associated with a significant reduction in larval viability as well as the differential expression of 21% of the genome (3109 genes). Among the genes identified were a set of genes previously known to be involved in xenobiotic metabolism, as well as genes involved in cellular energy production, oxidoreductase/carbohydrate metabolism, structural components and mRNA binding. Interestingly, of the 3109 genes whose expression was affected by host use, there was a significant overrepresentation of genes that lacked an orthologous call to the D. melanogaster genome, suggesting the possibility of an accelerated rate of evolution in these genes. Of the genes with a significant cactus effect, the majority, 2264 genes, did not exhibit a significant cactus-by-line interaction. This population-level approach facilitated the identification of genes involved in past cactus host shifts.

  3. Optimal conditions for production of extracellular protease from newly isolated Bacillus cereus strain CA15

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    Fikret Uyar

    2011-02-01

    Full Text Available An alkaline protease producer Bacillus sp. strain CA15 was isolated from soil. The microorganism was found to be closely related to Bacillus cereus based on 16S ribosomal DNA sequencing. The culture conditions for higher protease production were optimized with respect to carbon and nitrogen sources, metal ions, pH and temperature. Maximum protease production was obtained in the medium supplemented with 1% skim milk, 1% starch and 0.6% MgSO4.7H2O, initial pH 8.0 at 35oC. The best enzyme production was obtained during the stationary phase in which the cell density reached to 1.8x108 cells/mL. The level of protease was found to be low in the presence of inorganic nitrogen sources. The protease production was diminished in the presence of sucrose and lactose. The extreme stability towards Triton X-100, Tween 20 and SDS was observed by Bacillus sp. CA15 alkaline protease. The enzyme activity was inhibited by PMSF suggested that presence of serine residues at the active sites.

  4. Characterization of a highly toxic strain of Bacillus thuringiensis serovar kurstaki very similar to the HD-73 strain.

    Science.gov (United States)

    Reinoso-Pozo, Yaritza; Del Rincón-Castro, Ma Cristina; Ibarra, Jorge E

    2016-09-01

    The LBIT-1200 strain of Bacillus thuringiensis was recently isolated from soil, and showed a 6.4 and 9.5 increase in toxicity, against Manduca sexta and Trichoplusia ni, respectively, compared to HD-73. However, LBIT-1200 was still highly similar to HD-73, including the production of bipyramidal crystals containing only one protein of ∼130 000 kDa, its flagellin gene sequence related to the kurstaki serotype, plasmid and RepPCR patterns similar to HD-73, no production of β-exotoxin and no presence of VIP genes. Sequencing of its cry gene showed the presence of a cry1Ac-type gene with four amino acid differences, including two amino acid replacements in domain III, compared to Cry1Ac1, which may explain its higher toxicity. In conclusion, the LBIT-1200 strain is a variant of the HD-73 strain but shows a much higher toxicity, which makes this new strain an important candidate to be developed as a bioinsecticide, once it passes other tests, throughout its biotechnological development.

  5. Anthelmintic Effect of Bacillus thuringiensis Strains against the Gill Fish Trematode Centrocestus formosanus

    Science.gov (United States)

    Mendoza-Estrada, Luis Javier; Hernández-Velázquez, Víctor Manuel; Arenas-Sosa, Iván; Flores-Pérez, Fernando Iván; Morales-Montor, Jorge; Peña-Chora, Guadalupe

    2016-01-01

    Parasitic agents, such as helminths, are the most important biotic factors affecting aquaculture, and the fluke Centrocestus formosanus is considered to be highly pathogenic in various fish species. There have been efforts to control this parasite with chemical helminthicides, but these efforts have had unsuccessful results. We evaluated the anthelmintic effect of 37 strains of Bacillus thuringiensis against C. formosanus metacercariae in vitro using two concentrations of total protein, and only six strains produced high mortality. The virulence (CL50) on matacercariae of three strains was obtained: the GP308, GP526, and ME1 strains exhibited a LC50 of 146.2 μg/mL, 289.2 μg/mL, and 1721.9 μg/mL, respectively. Additionally, these six B. thuringiensis strains were evaluated against the cercariae of C. formosanus; the LC50 obtained from the GP526 strain with solubilized protein was 83.8 μg/mL, and it could be considered as an alternative control of the metacercariae and cercariae of this parasite in the productivity systems of ornamental fishes. PMID:27294137

  6. Isolation and Identification of a new Bacillus cereus strain and Characterization of its Neopullulanase

    Directory of Open Access Journals (Sweden)

    Soheila Davaeifar

    2015-03-01

    Full Text Available Identification and use of more efficient enzymes in the food and pharmaceutical industries is the focus of many researchers. The aim of this study was to search for a new bacterial strain capable of producing high levels of pullulanase applicable to biotechnology, the starch bioprocessing and food industries. A new pullulan hydrolyzing Bacillus strain was isolated and designated SDK2. Morphological and biochemical tests identified the strain as a putative Bacillus cereus strain, which was further characterized and confirmed through 16s rRNA sequencing, and was submitted to GeneBank, under the accession number FR6864500. Quantative analysis of the strain’s pullulanase activity was carried out by the Dintrosalicyclic (DNS acid-based assay. Thin layer chromatography (TLC of the culture supernatant, identified the extracellular pullulanase as neopullulanase. Effects of temperature and pH on pullulanase activity were also studied. The optimum conditions for enzyme activity, as represented by 60o C and a pH of 7, resulted in an activity of 13.43 U/ml, which is much higher than some of the previously reported activities. However, growth of B. cereus SDK2 was also observed at a pH range of 5 to 10, and temperatures of 30 oC to 50 oC. The effect of metal ions and reagents, such as Mg+2, Ca+2, Zn+2, Cu+2, Fe+2, Ni+2 on enzyme activity showed that Ca+2 ions increased pullulan activity, whereas the other ions and reagents inhibited pullulanase activity. The ability of B. cereus SDK2 to produce high levels of neopullulanase stable at 60 oC that can generate panose from pullulan, make this newly isolated strain a valuable source of debranching enzyme for biotechnology, the starch bioprocess and medical industries.

  7. Toxin production in a rare and genetically remote cluster of strains of the Bacillus cereus group

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    Granum Per

    2007-05-01

    Full Text Available Abstract Background Three enterotoxins are implicated in diarrhoeal food poisoning due to Bacillus cereus: Haemolysin BL (Hbl, Non-haemolytic enterotoxin (Nhe, and Cytotoxin K (CytK. Toxin gene profiling and assays for detection of toxin-producing stains have been used in attempts to evaluate the enterotoxic potential of B. cereus group strains. B. cereus strain NVH 391/98, isolated from a case of fatal enteritis, was genetically remote from other B. cereus group strains. This strain lacked the genes encoding Hbl and Nhe, but contains CytK-1. The high virulence of this strain is thought to be due to the greater cytotoxic activity of CytK-1 compared to CytK-2, and to a high level of cytK expression. To date, only three strains containing cytK-1 have been identified; B. cereus strains NVH 391/98, NVH 883/00, and INRA AF2. Results A novel gene variant encoding Nhe was identified in these three strains, which had an average of 80% identity in protein sequence with previously identified Nhe toxins. While culture supernatants containing CytK and Nhe from NVH 391/98 and INRA AF2 were highly cytotoxic, NVH 883/00 expressed little or no CytK and Nhe and was non-cytotoxic. Comparative sequence and expression studies indicated that neither the PlcR/PapR quorum sensing system, nor theYvrGH and YvfTU two-component systems, were responsible for the observed difference in toxin production. Additionally, phylogenetic analysis of 13 genes showed that NVH 391/98, NVH 883/00, and INRA AF2 comprise a novel cluster of strains genetically distant from other B. cereus group strains. Conclusion Due to its divergent sequence, the novel nhe operon had previously not been detected in NVH 391/98 using PCR and several monoclonal antibodies. Thus, toxigenic profiling based on the original nhe sequence will fail to detect the toxin in this group of strains. The observation that strain NVH 883/00 carries cytK-1 but is non-cytotoxic indicates that the detection of this gene

  8. Phylogenetic distribution of extracellular guanyl-preferring ribonucleases renews taxonomic status of two Bacillus strains.

    Science.gov (United States)

    Ulyanova, Vera; Shah Mahmud, Raihan; Dudkina, Elena; Vershinina, Valentina; Domann, Eugen; Ilinskaya, Olga

    2016-09-12

    The potential of microbial ribonucleases as promising antitumor and antiviral agents, determines today's directions of their study. One direction is connected with biodiversity of RNases. We have analyzed completed and drafted Bacillus genomes deposited in GenBank for the presence of coding regions similar to the gene of an extracellular guanyl-preferring RNase of Bacillus amyloliquefaciens (barnase). Orthologues of the barnase gene were detected in 9 species out of 83. All of these belong to "B. subtilis" group within the genus. B. subtilis itself, as well as some other species within this group, lack such types of RNases. RNases similar to barnase were also found in species of "B. cereus" group as a part of plasmid-encoded S-layer toxins. It was also found that taxonomic states of culture collection strains, which were initially described based on a limited set of phenotypic characteristics, can be misleading and need to be confirmed. Using several approaches such as matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), sequencing of genes for 16S ribosomal RNA and RNA polymerase subunit beta followed by reconstruction of phylogenetic trees, we have re-identified two RNase-secreting Bacillus strains: B. thuringiensis B-388 which should be assigned as B. altitudinis B388 and B. intermedius 7P which should be renamed as B. pumilus 7P. Therefore, small secreted guanyl-preferring RNases are the feature of "B. subtilis" group only, which is characterized by distinctive lifestyle and adaptation strategies to environment.

  9. Whole genome assembly of a natto production strain Bacillus subtilis natto from very short read data

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    Fujiyama Asao

    2010-04-01

    Full Text Available Abstract Background Bacillus subtilis natto is closely related to the laboratory standard strain B. subtilis Marburg 168, and functions as a starter for the production of the traditional Japanese food "natto" made from soybeans. Although re-sequencing whole genomes of several laboratory domesticated B. subtilis 168 derivatives has already been attempted using short read sequencing data, the assembly of the whole genome sequence of a closely related strain, B. subtilis natto, from very short read data is more challenging, particularly with our aim to assemble one fully connected scaffold from short reads around 35 bp in length. Results We applied a comparative genome assembly method, which combines de novo assembly and reference guided assembly, to one of the B. subtilis natto strains. We successfully assembled 28 scaffolds and managed to avoid substantial fragmentation. Completion of the assembly through long PCR experiments resulted in one connected scaffold for B. subtilis natto. Based on the assembled genome sequence, our orthologous gene analysis between natto BEST195 and Marburg 168 revealed that 82.4% of 4375 predicted genes in BEST195 are one-to-one orthologous to genes in 168, with two genes in-paralog, 3.2% are deleted in 168, 14.3% are inserted in BEST195, and 5.9% of genes present in 168 are deleted in BEST195. The natto genome contains the same alleles in the promoter region of degQ and the coding region of swrAA as the wild strain, RO-FF-1. These are specific for γ-PGA production ability, which is related to natto production. Further, the B. subtilis natto strain completely lacked a polyketide synthesis operon, disrupted the plipastatin production operon, and possesses previously unidentified transposases. Conclusions The determination of the whole genome sequence of Bacillus subtilis natto provided detailed analyses of a set of genes related to natto production, demonstrating the number and locations of insertion sequences that B

  10. Isolation of a new Mexican strain of Bacillus subtilis with antifungal and antibacterial activities.

    Science.gov (United States)

    Basurto-Cadena, M G L; Vázquez-Arista, M; García-Jiménez, J; Salcedo-Hernández, R; Bideshi, D K; Barboza-Corona, J E

    2012-01-01

    Although several strains of B. subtilis with antifungal activity have been isolated worldwide, to date there are no published reports regarding the isolation of a native B. subtilis strain from strawberry plants in Mexico. A native bacterium (Bacillus subtilis 21) demonstrated in vitro antagonistic activity against different plant pathogenic fungi. Under greenhouse conditions, it was shown that plants infected with Rhizoctonia solani and Fusarium verticillioides and treated with B. subtilis 21 produced augment in the number of leaves per plant and an increment in the length of healthy leaves in comparison with untreated plants. In addition, B. subtilis 21 showed activity against pathogenic bacteria. Secreted proteins by B. subtilis 21 were studied, detecting the presence of proteases and bacteriocin-like inhibitor substances that could be implicated in its antagonistic activity. Chitinases and zwittermicin production could not be detected. Then, B. subtilis 21 could potentially be used to control phytopathogenic fungi that infect strawberry plants.

  11. Isolation of a New Mexican Strain of Bacillus subtilis with Antifungal and Antibacterial Activities

    Directory of Open Access Journals (Sweden)

    M. G. L. Basurto-Cadena

    2012-01-01

    Full Text Available Although several strains of B. subtilis with antifungal activity have been isolated worldwide, to date there are no published reports regarding the isolation of a native B. subtilis strain from strawberry plants in Mexico. A native bacterium (Bacillus subtilis 21 demonstrated in vitro antagonistic activity against different plant pathogenic fungi. Under greenhouse conditions, it was shown that plants infected with Rhizoctonia solani and Fusarium verticillioides and treated with B. subtilis 21 produced augment in the number of leaves per plant and an increment in the length of healthy leaves in comparison with untreated plants. In addition, B. subtilis 21 showed activity against pathogenic bacteria. Secreted proteins by B. subtilis 21 were studied, detecting the presence of proteases and bacteriocin-like inhibitor substances that could be implicated in its antagonistic activity. Chitinases and zwittermicin production could not be detected. Then, B. subtilis 21 could potentially be used to control phytopathogenic fungi that infect strawberry plants.

  12. Complete Genome Sequence of a thermotolerant sporogenic lactic acid bacterium, Bacillus coagulans strain 36D1

    Energy Technology Data Exchange (ETDEWEB)

    Rhee, Mun Su [University of Florida, Gainesville; Moritz, Brelan E. [University of Florida, Gainesville; Xie, Gary [Los Alamos National Laboratory (LANL); Glavina Del Rio, Tijana [U.S. Department of Energy, Joint Genome Institute; Dalin, Eileen [U.S. Department of Energy, Joint Genome Institute; Tice, Hope [U.S. Department of Energy, Joint Genome Institute; Bruce, David [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Chertkov, Olga [Los Alamos National Laboratory (LANL); Brettin, Thomas S [ORNL; Han, Cliff [Los Alamos National Laboratory (LANL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Land, Miriam L [ORNL; Patel, Milind [University of Florida, Gainesville; Ou, Mark [University of Florida, Gainesville; Harbrucker, Roberta [University of Florida, Gainesville; Ingram, Lonnie O. [University of Florida; Shanmugam, Keelnathan T. [University of Florida

    2011-01-01

    Bacillus coagulans is a ubiquitous soil bacterium that grows at 50-55 C and pH 5.0 and fer- ments various sugars that constitute plant biomass to L (+)-lactic acid. The ability of this spo- rogenic lactic acid bacterium to grow at 50-55 C and pH 5.0 makes this organism an attrac- tive microbial biocatalyst for production of optically pure lactic acid at industrial scale not only from glucose derived from cellulose but also from xylose, a major constituent of hemi- cellulose. This bacterium is also considered as a potential probiotic. Complete genome se- quence of a representative strain, B. coagulans strain 36D1, is presented and discussed.

  13. Complete Genome Sequence of a thermotolerant sporogenic lactic acid bacterium, Bacillus coagulans strain 36D1

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Gary [Los Alamos National Laboratory (LANL); Dalin, Eileen [U.S. Department of Energy, Joint Genome Institute; Tice, Hope [U.S. Department of Energy, Joint Genome Institute; Chertkov, Olga [Los Alamos National Laboratory (LANL); Land, Miriam L [ORNL

    2011-01-01

    Bacillus coagulans is a ubiquitous soil bacterium that grows at 50-55 C and pH 5.0 and fer-ments various sugars that constitute plant biomass to L (+)-lactic acid. The ability of this sporogenic lactic acid bacterium to grow at 50-55 C and pH 5.0 makes this organism an attractive microbial biocatalyst for production of optically pure lactic acid at industrial scale not only from glucose derived from cellulose but also from xylose, a major constituent of hemi-cellulose. This bacterium is also considered as a potential probiotic. Complete genome squence of a representative strain, B. coagulans strain 36D1, is presented and discussed.

  14. The function of PlcR in Bacillus anthracis vaccine strain A16R.

    Science.gov (United States)

    Xiaolin, Jia; Dongshu, Wang; Zhiqi, Gao; Erling, Feng; Jiping, Zheng; Hengliang, Wang; Guiying, Guo; Xiankai, Liu

    2015-05-01

    Bacillus anthracis, B. thuringiensis and B. cereus are members of the B. cereus group. They share high genetic similarity. Whereas plcR (Phospholipase C regulator) usually encodes a functional pleiotropic activator protein in B. cereus and B. thuringiensis isolates, a characteristic nonsense mutation is found in all B. anthracis strains investigated, making the gene dysfunctional. To study the function of PlcR in B. anthracis, we used the B. cereus CMCC63301 genome as a template and constructed a recombinant expression plasmid pBE2A-plcR, and introduced it into the B. anthracis vaccine strain A16R, and then analyzed the activity of the hemolysin and sphingomyelinase. The results showed that transformation of B. anthracis with plasmid pBE2A-plcR carrying the native B. cereus plcR gene active the expression of sphingomyelinase gene, but did not activate expression of hemolysin genes of B. anthracis A16R.

  15. Construction and application of recombinant strain for the production of an alkaline protease from Bacillus licheniformis.

    Science.gov (United States)

    Lin, Songyi; Zhang, Meishuo; Liu, Jingbo; Jones, Gregory S

    2015-03-01

    The alkaline protease gene, Apr, from Bacillus licheniformis 2709 was cloned into an expression vector pET - 28b (+), to yield the recombinant plasmid pET-28b (+) - Apr. The pET-28b (+) - Apr was expressed in a high expression strain E. coli BL21. The amino acid sequence deduced from the DNA sequence analysis revealed a 98% identity to that of Bacillus licheniformis 2709. Sodium salt-Polyacrylamide gel electrophoresis (SDS-PAGE) was used to access the protein expression. SDS-PAGE analysis indicated a protein of Mr of 38.8 kDa. The medium components and condition of incubation were optimized for the growth state of a recombinant strain. The optimal composition of production medium was composed of glucose 8 g/L, peptone 8 g/L and salt solution 10 mL. The samples were incubated on a rotary shaker of 180 r/min at 37°C for 24 h.

  16. PENGENDALIAN JENTIK NYAMUK VEKTOR DEMAM BERDARAH, MALARIA DAN FILARIASIS MENGGUNAKAN STRAIN LOKAL BACILLUS THURINGIENSIS H-14

    Directory of Open Access Journals (Sweden)

    Blondie Ch. P.

    2012-09-01

    Full Text Available A study was conducted anticipating vector control for the control of Dengue Haemmoraghic Fever (Aedes aegypti, malaria (Anopheles aconitus and filariasis (Culex quinguefasciatus using a local strain of Bacillus thuringiensis H-14 grown in local media (coconut water and soybean infusion. Tryptose Phosphate Broth chemical media (standard media was used as a comparison to the media under investigation. Good growth was obtained in all media (local media and standard media and local strain of B. thuringiensis H-14 was effective against the three mosquito larvae. The local strain of Bacillus thuringiensis H-14 cultured in coconut water media, killed 50% of the third instar larvae of Ae. aegypti, An. aconitus and Cx. quinquefasciatus at concentrations of 13 x 10'5, 31 x 10-5 and 10 x 10-5 concentrations for 24 hours of exposure respectively and 13 x 10-5, 9 x 10-5 and 7 x 10-5 at 48 hours exposure. Meanwhile when B. thurigiensis H-14 were cultured in soybean infusion media, they killed 50% of the third instar larvae of Ae. aegypti, An. aconitus and Cx. quinquefasciatus at concentrations of 5 x 10-5, 7 x 10-5 and 4 x 10-5 for 24 hours of exposure respectively and 3 x 10-5, 5 x 10-5 and 3 x 10-5 at 48 hours exposure. However when they were cultured in TPB media, they killed 50% of the third instar larvae of Ae. aegypti, An. aconitus and Cx. quinquefasciatus at concentrations of 12 x 10-5, 15 x 10-5 and 3 x 10-5 for 24 hours of exposure respectively and 12 x 10-5, 15 x 10-5 and 2 x 10-5 at 48 hours exposure. This investigation shows that the local strain of B. thuringiensis H-14 has potential as bioinsecticide agent.

  17. Novel toxicity of Bacillus thuringiensis strains against the melon fruit fly, Bactrocera cucurbitae (Diptera: Tephritidae).

    Science.gov (United States)

    Shishir, Md Asaduzzaman; Akter, Asma; Bodiuzzaman, Md; Hossain, M Aftab; Alam, Md Musfiqul; Khan, Shakil Ahmed; Khan, Shakila Nargis; Hoq, M Mozammel

    2015-01-01

    Bactrocera cucurbitae (melon fruit fly) is one of the most detrimental vegetable-damaging pests in Bangladesh. The toxicity of Bacillus thuringiensis (Bt) has been reported against a few genera of Bactrocera in addition to numerous other insect species. Bt strains, harbouring cry1A-type genes were, therefore, assayed in vivo against the 3(rd) instar larvae of B. cucurbitae in this study. The biotype-based prevalence of cry1 and cry1A genes was calculated to be 30.8% and 11.16%, respectively, of the test strains (n=224) while their prevalence was greatest in biotype kurstaki. Though three indigenous Bt strains from biotype kurstaki with close genetic relationship exhibited higher toxicity, maximum mortalities were recorded for Btk HD-73 (96%) and the indigenous Bt JSc1 (93%). LC50 and LC99 values were determined to be 6.81 and 8.32 for Bt JSc1, 7.30 and 7.92 for Bt SSc2, and 6.99 and 7.67 for Btk HD-73, respectively. The cause of toxicity and its variation among the strains was found to be correlated with the synergistic toxic effects of cry1, cry2, cry3 and cry9 gene products, i.e. relevant Cry proteins. The novel toxicity of the B. thuringiensis strains against B. cucurbitae revealed in the present study thus will help in developing efficient and eco-friendly control measures such as Bt biopesticides and transgenic Bt cucurbits.

  18. Antimicrobial activity of a biosurfactant produced by Bacillus licheniformis strain M104 grown on whey

    Directory of Open Access Journals (Sweden)

    Eman Zakaria Gomaa

    2013-04-01

    Full Text Available The aim of the present study was to investigate the antimicrobial effect of the lipopeptide biosurfactants produced by Bacillus licheniformis strain M104 grown on whey. The biosurfactant was investigated for potential antimicrobial activity by using the disc-diffusion method against different Gram-positive bacteria {B subtilis, B. thuringiensis (two strains, B. cereus, Staphylococcus aureus (two strains and Listeria monocytogenes}, Gram-negative bacteria {(Pseudomonas aeruginosa, Escherichia coli (two strains, Salmonella typhimurium, Proteous vulgaris and Klebsiella pneumoniae and a yeast (Candida albicans}. The biosurfactant showed profoundly distinct antibacterial activity toward tested bacteria and displayed an antifungal activity against the tested yeast. Maximum antimicrobial activity of the biosurfactant was shown against S. aureus ATCC 25928. The biosurfactant had a broad inhibition effect on intracellular components of S. aureus ATCC 25928. The antimicrobial effect of lipopeptide biosurfactant produced by B. licheniformis strain M104 was time and concentration dependent. When biosurfactant was added to S. aureus medium in a concentration of (48 μg / ml, the maximum reduction of acid soluble phosphorous (53.06 %, total lipid (90.47 % total proteins (53.43%, RNA (83.29 % and DNA (48.50% were recorded after 12 h of incubation period. From the preliminary characterization results, it could be concluded that biosurfactants were a suitable alternative in potential applications of medical fields.

  19. Probiotic Bacillus cereus strains, a potential risk for public health in China

    Directory of Open Access Journals (Sweden)

    Kui eZhu

    2016-05-01

    Full Text Available Bacillus cereus is an important cause of foodborne infectious disease and food poisoning. However, B. cereus has also been used as a probiotic in human medicine and livestock production, with low standards of safety assessment. In this study, we evaluated the safety of 15 commercial probiotic B. cereus preparations from China in terms of mislabeling, toxin production, and transferable antimicrobial resistance. Most preparations were incorrectly labeled, as they contained additional bacterial species; one product did not contain viable B. cereus at all. In total, 18 B. cereus group strains – specifically B. cereus and B. thuringiensis – were isolated. Enterotoxin genes nhe, hbl, and cytK1, as well as the ces-gene were assessed by PCR. Enterotoxin production and cytotoxicity were confirmed by ELISA and cell culture assays, respectively. All isolated B. cereus group strains produced the enterotoxin Nhe; 15 strains additionally produced Hbl. Antimicrobial resistance was assessed by microdilution; resistance genes were detected by PCR and further characterized by sequencing, transformation and conjugation assays. Nearly half of the strains harbored the antimicrobial resistance gene tet(45. In one strain, tet(45 was situated on a mobile genetic element – encoding a site specific recombination mechanism – and was transferable to Staphylococcus aureus and B. subtilis by electro-transformation. In view of the wide and uncontrolled use of these products, stricter regulations for safety assessment, including determination of virulence factors and transferable antimicrobial resistance genes, are urgently needed.

  20. [Probiotic features of carotene producing strains Bacillus sp. 1.1 and B. amyloliquefaciens UCM B-5113].

    Science.gov (United States)

    Avdeeva, L V; Nechypurenko, O O; Kharhota, M A

    2015-01-01

    Researched probiotic properties of carotinproducing strains Bacillus sp. 1.1 and B. amyloliquefaciens UCM B-5113. It was established that Bacillus sp. 1.1 characterized by high and middle antagonistic activity against museums and actual test cultures and B. amyloliquefaciens UCM B-5113 shown middle and low activity. They grew up and formed a pigment at pH 6.0 in the presence of 0.4% bile. Bacillus sp. 1.1 and B. amyloliquefaciens UCM B-5113 were avirulent, had low antagonistic activity and characterized by susceptibility to antimicrobial agents, excluding colistin. The results suggested the possibility to create based on Bacillus sp. 1.1 and B. amyloliquefaciens UCM B-5113 probiotic preparation.

  1. Characterization of antagonistic-potential of two Bacillus strains and their biocontrol activity against Rhizoctonia solani in tomato.

    Science.gov (United States)

    Solanki, Manoj Kumar; Singh, Rajesh Kumar; Srivastava, Supriya; Kumar, Sudheer; Kashyap, Prem Lal; Srivastava, Alok K

    2015-01-01

    To investigate the biocontrol mechanism of two antagonistic Bacillus strains (Bacillus subtilis MB14 and Bacillus amyloliquefaciens MB101), three in vitro antagonism assays were screened and the results were concluded that both strains inhibited Rhizoctonia solani growth in a similar manner by dual culture assay, but the maximum percent of inhibition only resulted with MB101 by volatile and diffusible metabolite assays. Moreover, cell free supernatant (CFS) of MB101 also showed significant (p > 0.05) growth inhibition as compared to MB14, when 10 and 20% CFS mix with the growth medium of R. solani. After in vitro-validation, both strains were evaluated under greenhouse and the results concluded that strain MB101 had significant biocontrol potential as compared to MB14. Strain MB101 was enhanced the plant height, biomass and chlorophyll content of tomato plant through a higher degree of root colonization. In field trials, strain MB101 showed higher lessening in root rot symptoms with significant fruit yield as compare to strain MB14 and infected control. Next to the field study, the presence of four antibiotic genes (srfAA, fenD, ituC, and bmyB) also concluded the antifungal nature of both Bacillus strains. Phylogenetic analysis of protein sequences revealed a close relatedness of three genes (srfAA, fenD, and ituC) with earlier reported sequences of B. subtilis and B. amyloliquefaciens. However, bmyB showed heterogeneity in among both strains (MB14 and MB101) and it may be concluded that higher degree of antagonism, root colonization and different antibiotic producing genes may play an important role in biocontrol mechanism of strain MB101.

  2. Bacillus thuringiensis monogenic strains: screening and interactions with insecticides used against rice pests

    Directory of Open Access Journals (Sweden)

    Laura M.N. Pinto

    2012-06-01

    Full Text Available The screening of Bacillus thuringiensis (Bt Cry proteins with high potential to control insect pests has been the goal of numerous research groups. In this study, we evaluated six monogenic Bt strains (Bt dendrolimus HD-37, Bt kurstaki HD-1, Bt kurstaki HD-73, Bt thuringiensis 4412, Bt kurstaki NRD-12 and Bt entomocidus 60.5, which codify the cry1Aa, cry1Ab, cry1Ac, cry1Ba, cry1C, cry2A genes respectively as potential insecticides for the most important insect pests of irrigated rice: Spodoptera frugiperda, Diatraea saccharalis, Oryzophagus oryzae, Oebalus poecilus and Tibraca limbativentris. We also analyzed their compatibility with chemical insecticides (thiamethoxam, labdacyhalothrin, malathion and fipronil, which are extensively used in rice crops. The bioassay results showed that Bt thuringiensis 4412 and Bt entomocidus 60.5 were the most toxic for the lepidopterans, with a 93% and 82% mortality rate for S. frugiperda and D. saccharalis, respectively. For O. oryzae, the Bt kurstaki NRD-12 (64% and Bt dendrolimus HD-37 (62% strains were the most toxic. The Bt dendrolimus HD-37 strain also caused high mortality (82% to O. poecilus, however the strains assessed to T. limbativentris caused a maximum rate of 5%. The assays for the Bt strains interaction with insecticides revealed the compatibility of the six strains with the four insecticides tested. The results from this study showed the high potential of cry1Aa and cry1Ba genes for genetic engineering of rice plants or the strains to biopesticide formulations.

  3. Alveolar macrophages infected with Ames or Sterne strain of Bacillus anthracis elicit differential molecular expression patterns.

    Directory of Open Access Journals (Sweden)

    Felicia D Langel

    Full Text Available Alveolar macrophages (AMs phagocytose Bacillus anthracis following inhalation and induce the production of pro-inflammatory cytokines and chemokines to mediate the activation of innate immunity. Ames, the virulent strain of B. anthracis, contains two plasmids that encode the antiphagocytic poly-γ-d-glutamic acid capsule and the lethal toxin. The attenuated Sterne strain of B. anthracis, which lacks the plasmid encoding capsule, is widely adapted as a vaccine strain. Although differences in the outcome of infection with the two strains may have originated from the presence or absence of an anti-phagocytic capsule, the disease pathogenesis following infection will be manifested via the host responses, which is not well understood. To gain understanding of the host responses at cellular level, a microarray analysis was performed using primary rhesus macaque AMs infected with either Ames or Sterne spores. Notably, 528 human orthologs were identified to be differentially expressed in AMs infected with either strain of the B. anthracis. Meta-analyses revealed genes differentially expressed in response to B. anthracis infection were also induced upon infections with multiple pathogens such as Francisella Novicida or Staphylococcus aureus. This suggests the existence of a common molecular signature in response to pathogen infections. Importantly, the microarray and protein expression data for certain cytokines, chemokines and host factors provide further insights on how cellular processes such as innate immune sensing pathways, anti-apoptosis versus apoptosis may be differentially modulated in response to the virulent or vaccine strain of B. anthracis. The reported differences may account for the marked difference in pathogenicity between these two strains.

  4. [Screening of Bacillus thuringiensis strains containing vip3A genes and analysis of gene conservation].

    Science.gov (United States)

    Chen, Jian-Wu; Tang, Li-Xia; Song, Shao-Yun; Yuan, Mei-Jin; Pang, Yi

    2003-09-01

    Vip3A, a novel insecticidal protein, is secreted by Bacillus thuringiensis (Bt) during vegetative growth. Vip3A protein possesses insecticidal activity against a wild spectrum of lepidopteran insect larvae. Since the first cloning of vip3A gene from Bt, many other vip3A genes have been isolated. To investigate vip3A genes contribution to Bt and reflect the revolution relationships, the strains containing vip3A genes were screened and gene similarity was analyzed. 114 wild-type Bacillus thuringiensis (Bt) strains isolated from different regions and 41 standard Bt strains from the Institute of Pasteur were screened for the vip3A genes using PCR amplification. 39 strains including B. thuringiensis subsp. kurstaki (Btk) HD-1 were found to contain the vip3A genes. Because acrystallerous strain Cry- B derived from Btk HD-1 was proved not to contain vip3A gene, it suppose that the vip3A gene may be located at the plasmids. Vip3A proteins expressed in these strains were detected with polyclonal antibody by Western blot and 4 strains among them were shown not to express the Vip3A proteins. The vip3A genes amplified from wild-type Bacillus thuringiensis strains S101 and 611 with different levels of activity against lepidopteran insect larvae were cloned into pGEM-T Easy vector. Alignment of these 2 putative Vip3A proteins with 6 others (Vip3A (a), Vip3A(b), Vip3A-S, Vip3A-S184, Vip83 and Vip3V) in the GenBank data base and 2 reported Vip3A proteins (Vip14 and Vip15) showed that vip3A genes are highly conservative. The plasmids pOTP-S101 and pOTP-611 were constructed by in- serting 2 vip3A genes (vip3A-S101 and vip3A-611) into the expression vector pQE30 respectively and were transformed into E. coli M15. E. coli M15 cells harboring the pOTP plasmids were induced with 1 mmol/L IPTG to express 89 kDa protein. Experiments showed that the level of soluble proteins of Vip3A-S101 in E. coli M15[pOTP-S101] and Vip3A-611 in E. coli M15 [pOTP-611] were about 48% and 35% respectively

  5. Draft Genome Sequences of Type Strains Bacillus drentensis DSM 15600T and Bacillus novalis DSM 15603T.

    Science.gov (United States)

    Liu, Bo; Liu, Guo-Hong; Zhu, Yu-Jing; Wang, Jie-Ping; Che, Jian-Mei; Chen, Qian-Qian; Chen, Zheng

    2016-12-15

    Here, we report the draft genome sequences of Bacillus drentensis DSM 15600(T) and Bacillus novalis DSM 15603(T) with 5,305,306 bp and 5,667,584 bp, respectively, which will provide useful information for the functional gene mining and application of these two species. The average DNA G+C contents were 38.91% and 40.01%, respectively.

  6. ISOLATION AND CHARACTERIZATION OF BIFENTHRIN CATABOLIZING BACTERIAL STRAIN BACILLUS CIBI FROM SOIL FOR PYRETHROIDS BIODEGRADATION

    Directory of Open Access Journals (Sweden)

    Preeti Pandey

    2014-01-01

    Full Text Available Pyrethroids are commonly used in most parts of the world and are reported to have potential health risks. Bifenthrin, a third generation pyrethroid used as insecticide has caused potential effect on aquatic life and human health. Bioremediation is a practical approach to reduce pesticide in the environment and reports of microbial degradation of bifenthrin are meagre. This study was aimed at isolating and characterizing bacterial isolates for the efficient removal of bifenthrin residues in the environment. A bacterial strain PGS-4 isolated from sewage of pesticide industry was tested for growth at higher concentration of bifenthrin (800 mg L-1 and the optimum pH and temperature were determined. The strain utilized bifenthrin as sole carbon source for growth over a wide range of pH (4.0-9.0 and temperatures (16-37°C. On the basis of growth kinetics studies, the optimal conditions were determined to be pH 7.0-8.0 and 30°C. 16S rRNA gene sequence analysis showed that strain PGS-4 forms a distinct phylogenetic lineage within the evolutionary radiation encompassed by the genus Bacillus and showed 99% similarity to that of Bacillus cibi. This study depicts the ability of B. cibi to utilize bifenthrin at higher concentration under in vitro thereby can be used in eliminating bifenthrin from contaminated soils as a practical approach to reduce pyrethroid toxicity in the environment.

  7. Pathway and kinetics of cyhalothrin biodegradation by Bacillus thuringiensis strain ZS-19

    Science.gov (United States)

    Chen, Shaohua; Deng, Yinyue; Chang, Changqing; Lee, Jasmine; Cheng, Yingying; Cui, Zining; Zhou, Jianuan; He, Fei; Hu, Meiying; Zhang, Lian-Hui

    2015-01-01

    Cyhalothrin is a common environmental pollutant which poses increased risks to non-target organisms including human beings. This study reported for the first time a newly isolated strain, Bacillus thuringiensis ZS-19 completely degraded cyhalothrin in minimal medium within 72 h. The bacterium transformed cyhalothrin by cleavage of both the ester linkage and diaryl bond to yield six intermediate products. Moreover, a novel degradation pathway of cyhalothrin in strain ZS-19 was proposed on the basis of the identified metabolites. In addition to degradation of cyhalothrin, this strain was found to be capable of degrading 3-phenoxybenzoic acid, a common metabolite of pyrethroids. Furthermore, strain ZS-19 participated in efficient degradation of a wide range of pyrethroids including cyhalothrin, fenpropathrinn, deltamethrin, beta-cypermethrin, cyfluthrin and bifenthrin. Taken together, our results provide insights into the mechanism of cyhalothrin degradation and also highlight the promising potentials of B.thuringiensis ZS-19 in bioremediation of pyrethroid-contaminated environment. This is the first report of (i) degradation of cyhalothrin and other pyrethroids by B.thuringiensis, (ii) identification of 3-phenoxyphenyl acetonitrile and N-(2-isoproxy-phenyl)-4-phenoxy-benzamide as the metabolites in the degradation pathway of pyrethroids, and (iii) a pathway of degradation of cyhalothrin by cleavage of both the ester linkage and diaryl bond in a microorganism. PMID:25740758

  8. Pathway and kinetics of cyhalothrin biodegradation by Bacillus thuringiensis strain ZS-19.

    Science.gov (United States)

    Chen, Shaohua; Deng, Yinyue; Chang, Changqing; Lee, Jasmine; Cheng, Yingying; Cui, Zining; Zhou, Jianuan; He, Fei; Hu, Meiying; Zhang, Lian-Hui

    2015-03-05

    Cyhalothrin is a common environmental pollutant which poses increased risks to non-target organisms including human beings. This study reported for the first time a newly isolated strain, Bacillus thuringiensis ZS-19 completely degraded cyhalothrin in minimal medium within 72 h. The bacterium transformed cyhalothrin by cleavage of both the ester linkage and diaryl bond to yield six intermediate products. Moreover, a novel degradation pathway of cyhalothrin in strain ZS-19 was proposed on the basis of the identified metabolites. In addition to degradation of cyhalothrin, this strain was found to be capable of degrading 3-phenoxybenzoic acid, a common metabolite of pyrethroids. Furthermore, strain ZS-19 participated in efficient degradation of a wide range of pyrethroids including cyhalothrin, fenpropathrinn, deltamethrin, beta-cypermethrin, cyfluthrin and bifenthrin. Taken together, our results provide insights into the mechanism of cyhalothrin degradation and also highlight the promising potentials of B.thuringiensis ZS-19 in bioremediation of pyrethroid-contaminated environment. This is the first report of (i) degradation of cyhalothrin and other pyrethroids by B.thuringiensis, (ii) identification of 3-phenoxyphenyl acetonitrile and N-(2-isoproxy-phenyl)-4-phenoxy-benzamide as the metabolites in the degradation pathway of pyrethroids, and (iii) a pathway of degradation of cyhalothrin by cleavage of both the ester linkage and diaryl bond in a microorganism.

  9. Mining rare and ubiquitous toxin genes from a large collection of Bacillus thuringiensis strains.

    Science.gov (United States)

    Li, Ying; Shu, Changlong; Zhang, Xuewen; Crickmore, Neil; Liang, Gemei; Jiang, Xingfu; Liu, Rongmei; Song, Fuping; Zhang, Jie

    2014-10-01

    There has been considerable effort made in recent years for research groups and other organizations to build up large collections of strains of Bacillus thuringiensis in the search for genes encoding novel insecticidal toxins, or encoding novel metabolic pathways. Whilst next generation sequencing allows the detailed genetic characterization of a bacterial strain with relative ease it is still not practicable for large strain collections. In this work we assess the practicability of mining a mixture of genomic DNA from a two thousand strain collection for particular genes. Using PCR the collection was screened for both a rare (cry15) toxin gene as well as a more commonly found gene (vip3A). The method was successful in identifying both a cry15 gene and multiple examples of the vip3A gene family including a novel member of this family (vip3Aj). A number of variants of vip3Ag were cloned and expressed, and differences in toxicity observed despite extremely high sequence similarity.

  10. Keratinase production and keratin degradation by a mutant strain of Bacillus subtilis

    Institute of Scientific and Technical Information of China (English)

    Cheng-gang CAI; Bing-gan LOU; Xiao-dong ZHENG

    2008-01-01

    A new feather-degrading bacterium was isolated from a local feather waste site and identified as Bacillus subtilis based on morphological, physiochemical, and phylogenetic characteristics. Screening for mutants with elevated keratinolytic activity using N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis resulted in a mutant strain KD-N2 producing keratinolytic activity about 2.5 times that of the wild-type strain. The mutant strain produced inducible keratinase in different substrates of feathers, hair, wool and silk under submerged cultivation. Scanning electron microscopy studies showed the degradation of feathers, hair and silk by the keratinase. The optimal conditions for keratinase production include initial pH of 7.5, inoculum size of 2% (v/v), age of inoculum of 16 h, and cultivation at 23 ℃. The maximum keratinolytic activity of KD-N2 was achieved after 30 h. Essential amino acids like threonine, valine, methionine as well as ammonia were produced when feathers were used as substrates. Strain KD-N2,therefore, shows great promise of finding potential applications in keratin hydrolysis and keratinase production.

  11. Genomic and chemical insights into biosurfactant production by the mangrove-derived strain Bacillus safensis CCMA-560.

    Science.gov (United States)

    Domingos, Daniela Ferreira; de Faria, Andreia Fonseca; de Souza Galaverna, Renan; Eberlin, Marcos Nogueira; Greenfield, Paul; Zucchi, Tiago Domingues; Melo, Itamar Soares; Tran-Dinh, Nai; Midgley, David; de Oliveira, Valéria Maia

    2015-04-01

    Many Bacillus species can produce biosurfactant, although most of the studies on lipopeptide production by this genus have been focused on Bacillus subtilis. Surfactants are broadly used in pharmaceutical, food and petroleum industry, and biological surfactant shows some advantages over the chemical surfactants, such as less toxicity, production from renewable, cheaper feedstocks and development of novel recombinant hyperproducer strains. This study is aimed to unveil the biosurfactant metabolic pathway and chemical composition in Bacillus safensis strain CCMA-560. The whole genome of the CCMA-560 strain was previously sequenced, and with the aid of bioinformatics tools, its biosurfactant metabolic pathway was compared to other pathways of closely related species. Fourier transform infrared (FTIR) and high-resolution TOF mass spectrometry (MS) were used to characterize the biosurfactant molecule. B. safensis CCMA-560 metabolic pathway is similar to other Bacillus species; however, some differences in amino acid incorporation were observed, and chemical analyses corroborated the genetic results. The strain CCMA-560 harbours two genes flanked by srfAC and srfAD not present in other Bacillus spp., which can be involved in the production of the analogue gramicidin. FTIR and MS showed that B. safensis CCMA-560 produces a mixture of at least four lipopeptides with seven amino acids incorporated and a fatty acid chain with 14 carbons, which makes this molecule similar to the biosurfactant of Bacillus pumilus, namely, pumilacidin. This is the first report on the biosurfactant production by B. safensis, encompassing the investigation of the metabolic pathway and chemical characterization of the biosurfactant molecule.

  12. Augmentation of tribenuron methyl removal from polluted soil with Bacillus sp.strain BS2 and indigenous earthworms

    Institute of Scientific and Technical Information of China (English)

    Qiang Tang; Zhiping Zhao; Yajun Liu; Nanxi Wang; Baojun Wang; Yanan Wang; Ningyi Zhou; Shuangjiang Liu

    2012-01-01

    Tribenuron methyl(TBM)is a member of the sulfonylurea herbicide family and is widely used worldwide.In this study,TBMdegrading bacteria were enriched with TBM as potential carbon,nitrogen or sulfur source,and 44 bacterial isolates were obtained.These isolates were phylogenetically diverse,and were grouped into 25 operational taxonomic units and 14 currently known genera.Three representatives,Bacillus sp.strain BS2,Microbacterium sp.strain BS3,and Cellulosimicrobium sp.strain BS 11,were selected,and their growth and TBM removal from culture broth were investigated.In addition,indigenous earthworms were collected and applied to augment TBM degradation in lab-scale soil column experiments.Results demonstrated that Bacillus sp.strain BS2 and earthworms significantly increased TBM removal during soil column experiments.

  13. Biodegradation of hexavalent chromium (Cr+6) in wastewater using Pseudomonas sp. and Bacillus sp. bacterial strains

    Energy Technology Data Exchange (ETDEWEB)

    Qasim, Muhammad [Department of Chemical Engineering, American University of Sharjah (United Arab Emirates)

    2013-07-01

    The recovery of toxic metal compounds is a deep concern in all industries. Hexavalent chromium is particularly worrying because of its toxic influence on human health. In this paper, biodegradation of hexavalent chromium (Cr+6) present in wastewater has been studied using two different bacterial strains; Pseudomonas sp. and Bacillus sp. A chemostat (with and without recycle of cells) with 10 L liquid culture volume was used to study the substrate and the biomass cell concentrations with time. Also, the degree of substrate conversion was studied by the varying the dilution rate as an independent parameter. The dilution rate (ratio of feed flow rate to the culture volume) was varied by varying the feed volumetric rate from 110-170 mL/h for inlet hexavalent chromium concentrations of 70 mg/dm3. The results show that a chemostat with recycle gives a better performance in terms of substrate conversion than a chemostat without a recycle. Moreover, the degree of substrate conversion decreases as the dilution rate is increased. Also, Bacillus sp. was found to give higher conversions compared to pseudomonas sp.

  14. Genome Sequence of Bacillus sp. Strain UMTAT18 Isolated from the Dinoflagellate Alexandrium tamiyavanichii Found in the Straits of Malacca

    Science.gov (United States)

    Ming, Gan Han; Mohd Noor, Mohd Ezhar; Sung, Yeong Yik; Usup, Gires

    2016-01-01

    Bacillus sp. strain UMTAT18 was isolated from the harmful dinoflagellate Alexandrium tamiyavanichii. Its genome consists of 5,479,367 bp with 5,546 open reading frames, 102 tRNAs, and 29 rRNAs. Gene clusters for biosynthesis of nonribosomal peptides, bacteriocin, and lantipeptide were identified. It also contains siderophore and genes related to stress tolerance.

  15. Complete genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium of Calendula officinalis

    Energy Technology Data Exchange (ETDEWEB)

    Koeberl, Martina; White, Richard A.; Erschen, Sabine; Spanberger, Nora; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele

    2015-08-13

    The genome sequence of Bacillus amyloliquefaciens strain Co1-6, a plant growth-promoting rhizobacterium (PGPR) with broad-spectrum antagonistic activities against plant pathogenic fungi, bacteria and nematodes, consists of a single 3.9 Mb circular chromosome. The genome reveals genes putatively responsible for its promising biocontrol and PGP properties.

  16. Analysis of germination capacity and germinant receptor (sub)clusters of genomesequenced Bacillus cereus environmental isolates and model strains

    NARCIS (Netherlands)

    Warda, Alicja K.; Xiao, Yinghua; Boekhorst, Jos; Wells-Bennik, Marjon H.J.; Nierop Groot, Masja N.; Abee, Tjakko

    2017-01-01

    Spore germination of 17 Bacillus cereus food isolates and reference strains was evaluated using flow cytometry analysis in combination with fluorescent staining at a single-spore level. This approach allowed for rapid collection of germination data under more than 20 conditions, including heat ac

  17. Draft Genome Sequence of Bacillus licheniformis Strain YNP1-TSU Isolated from Whiterock Springs in Yellowstone National Park.

    Science.gov (United States)

    O'Hair, Joshua A; Li, Hui; Thapa, Santosh; Scholz, Matthew B; Zhou, Suping

    2017-03-02

    Novel cellulolytic microorganisms can potentially influence second-generation biofuel production. This paper reports the draft genome sequence of Bacillus licheniformis strain YNP1-TSU, isolated from hydrothermal-vegetative microbiomes inside Yellowstone National Park. The assembled sequence contigs predicted 4,230 coding genes, 66 tRNAs, and 10 rRNAs through automated annotation.

  18. Genome Sequence of Historical Bacillus anthracis Strain Tyrol 4675 Isolated from a Bovine Anthrax Case in Austria

    Science.gov (United States)

    Antwerpen, Markus; Wölfel, Roman

    2017-01-01

    ABSTRACT In 1988, an outbreak of anthrax occurred among cattle in the Austrian state of Tyrol. Since then, Austria has been declared anthrax-free. Here, we report the draft genome sequence of one of these last outbreak strains, Bacillus anthracis Tyrol 4675, isolated from a diseased cow. PMID:28280006

  19. Finished Genome Sequence of Bacillus cereus Strain 03BB87, a Clinical Isolate with B. anthracis Virulence Genes.

    Science.gov (United States)

    Johnson, Shannon L; Minogue, Timothy D; Teshima, Hazuki; Davenport, Karen W; Shea, April A; Miner, Haven L; Wolcott, Mark J; Chain, Patrick S G

    2015-01-15

    Bacillus cereus strain 03BB87, a blood culture isolate, originated in a 56-year-old male muller operator with a fatal case of pneumonia in 2003. Here we present the finished genome sequence of that pathogen, including a 5.46-Mb chromosome and two plasmids (209 and 52 Kb, respectively).

  20. Draft Genome Sequence of Bacillus licheniformis Strain GB2, a Hydrocarbon-Degrading and Plant Growth-Promoting Soil Bacterium.

    Science.gov (United States)

    Gkorezis, Panagiotis; Van Hamme, Jonathan; Bottos, Eric; Thijs, Sofie; Balseiro-Romero, Maria; Monterroso, Carmela; Kidd, Petra Suzan; Rineau, Francois; Weyens, Nele; Sillen, Wouter; Vangronsveld, Jaco

    2016-06-23

    We report the 4.39 Mb draft genome of Bacillus licheniformis GB2, a hydrocarbonoclastic Gram-positive bacterium of the family Bacillaceae, isolated from diesel-contaminated soil at the Ford Motor Company site in Genk, Belgium. Strain GB2 is an effective plant-growth promoter useful for diesel fuel remediation applications based on plant-bacterium associations.

  1. Draft Genome Sequence of Bacillus licheniformis Strain YNP1-TSU Isolated from Whiterock Springs in Yellowstone National Park

    Science.gov (United States)

    O'Hair, Joshua A.; Li, Hui; Thapa, Santosh; Scholz, Matthew B.

    2017-01-01

    ABSTRACT Novel cellulolytic microorganisms can potentially influence second-generation biofuel production. This paper reports the draft genome sequence of Bacillus licheniformis strain YNP1-TSU, isolated from hydrothermal-vegetative microbiomes inside Yellowstone National Park. The assembled sequence contigs predicted 4,230 coding genes, 66 tRNAs, and 10 rRNAs through automated annotation. PMID:28254968

  2. Preliminary Study and Improve the Production of Metabolites with Antifungal Activity by A Bacillus Sp Strain IBA 33

    Directory of Open Access Journals (Sweden)

    María Antonieta Gordillo

    2009-04-01

    Full Text Available Bacillus sp strain IBA 33 metabolites, isolated from decaying lemon fruits, were evaluated for the control of pathogenic and non-pathogenic fungi (Penicillium digitatum, Geotrichum candidum, Penicillium expansum, Aspergillus clavatus, Aspergillus flavus, Aspergillus niger, and Fusarium moniliforme. These metabolites were recovered from Landy medium (LM without aminoacids. In order to optimize metabolites production the LM was modified by adding different concentrations and sources of amino acids and carbohydrates at different culture conditions. Bacillus sp strain IBA 33 metabolites efficacy to control fungi were evaluated with in vitro and in vivo assays. A. flavus growth inhibition was 52% with the metabolites of Bacillus sp strain IBA 33 recovered from LM (MBLM in vitro assays. MBLM supplemented with 0.5% glutamic acid, inhibited the growth of P. digitatum, G. candidum, A. clavatus, A. niger and F. moniliforme by 65%, 88.44%, 84%, 34% and 92% respectively. The highest inhibition of P. expansum was 45% with MBLM supplemented with 0.5% aspartic acid. Similar results were obtained in vivo assays. These results showed that Bacillus sp strain IBA 33 metabolites specificity against fungi depended on the composition of the LM.

  3. Preliminary Study and Improve the Production of Metabolites with Antifungal Activity by A Bacillus Sp Strain IBA 33

    Directory of Open Access Journals (Sweden)

    María Antonieta Gordillo

    2009-01-01

    Full Text Available Bacillus sp strain IBA 33 metabolites, isolated from decaying lemon fruits, were evaluated for the control of pathogenic and non-pathogenic fungi (Penicillium digitatum, Geotrichum candidum, Penicillium expansum, Aspergillus clavatus, Aspergillus flavus, Aspergillus niger, and Fusarium moniliforme. These metabolites were recovered from Landy medium (LM without aminoacids. In order to optimize metabolites production the LM was modified by adding different concentrations and sources of amino acids and carbohydrates at different culture conditions.Bacillus sp strain IBA 33 metabolites efficacy to control fungi were evaluated with in vitro and in vivo assays. A. flavus growth inhibition was 52% with the metabolites of Bacillus sp strain IBA 33 recovered from LM (MBLM in vitro assays. MBLM supplemented with 0.5% glutamic acid, inhibited the growth of P. digitatum, G. candidum, A. clavatus, A. niger and F. moniliforme by 65%, 88.44%, 84%, 34% and 92% respectively. The highest inhibition of P. expansum was 45% with MBLM supplemented with 0.5% aspartic acid. Similar results were obtained in vivo assays. These results showed that Bacillus sp strain IBA 33 metabolites specificity against fungi depended on the composition of the LM.

  4. Modified Bacillus thuringiensis toxins and a hybrid B. thuringiensis strain counter greenhouse-selected resistance in Trichoplusia ni.

    Science.gov (United States)

    Franklin, Michelle T; Nieman, Christal L; Janmaat, Alida F; Soberón, Mario; Bravo, Alejandra; Tabashnik, Bruce E; Myers, Judith H

    2009-09-01

    Resistance of greenhouse-selected strains of the cabbage looper, Trichoplusia ni, to Bacillus thuringiensis subsp. kurstaki was countered by a hybrid strain of B. thuringiensis and genetically modified toxins Cry1AbMod and Cry1AcMod, which lack helix alpha-1. Resistance to Cry1AbMod and Cry1AcMod was >100-fold less than resistance to native toxins Cry1Ab and Cry1Ac.

  5. Draft Genome Sequence of Bacillus thuringiensis Serovar Tolworthi Strain Na205-3, an Isolate Toxic for Helicoverpa armigera

    Science.gov (United States)

    Palma, Leopoldo; Muñoz, Delia; Murillo, Jesús

    2014-01-01

    We report here the complete annotated 6,510,053-bp draft genome sequence of Bacillus thuringiensis serovar tolworthi strain Na205-3, which is toxic for Helicoverpa armigera. This strain potentially contains nine insecticidal toxin genes homologous to cry1Aa12, cry1Ab1, cry1Ab8, cry1Ba1, cry1Af1, cry1Ia10, vip1Bb1, vip2Ba2, and vip3Aa6. PMID:24625875

  6. Purification and characterization of a moderately thermostable xylanase from Bacillus sp. strain SPS-0.

    Science.gov (United States)

    Bataillon; Nunes Cardinali A; Castillon; Duchiron

    2000-02-01

    A Bacillus spp. strain SPS-0, isolated from a hot spring in Portugal, produced an extracellular xylanase upon growth on wheat bran arabinoxylan. The enzyme was purified to homogeneity by ammonium sulfate precipitation, anion exchange, gel filtration, and affinity chromatography. The optimum temperature and pH for activity was 75 degrees C and 6.0. Xylanase was stable up to 70 degrees C for 4 h at pH 6.0 in the presence of xylane. Xylanase was completely inhibited by the Hg(2+) ions. beta-Mercaptoethanol, dithiothreitol, and Mn(2+) stimulated the xylanase activity. The products of birchwood xylan hydrolysis were xylose, xylobiose, xylotriose, and xylotetraose. Kinetic experiments at 60 degrees C and pH 6.0 gave V(max) and K(m)values of 2420 nkat/mg and 0.7 mg/ml.

  7. Metabolic flexibility of d-ribose producer strain of Bacillus pumilus under environmental perturbations

    DEFF Research Database (Denmark)

    Srivastava, Rajesh K.; Maiti, Soumen K.; Das, Debasish;

    2012-01-01

    The metabolic reaction rate vector is a bridge that links gene and protein expression alterations to the phenotypic endpoint. We present a simple approach for the estimation of flux distribution at key branch points in the metabolic network by using substrate uptake, metabolite secretion rate......, and biomass growth rate for transketolase (tkt) deficient Bacillus pumilus ATCC 21951. We find that the glucose-6-phosphate (G6P) and pseudo catabolic/anabolic branch points are flexible in the d-ribose-producing tkt deficient strain of B. pumilus. The normalized flux through the pentose phosphate pathway...... (PPP) varied from 1.5 to 86 % under different growth conditions, thereby enabling substantial extracellular accumulation of d-ribose under certain conditions. Interestingly, the flux through PPP was affected by the extracellular phosphate concentration and dissolved oxygen concentration. This metabolic...

  8. Biochemical characterisation of lipase from a new strain of Bacillus sp. ITP-001

    Directory of Open Access Journals (Sweden)

    José Murillo P. Barbosa

    2012-01-01

    Full Text Available Lipases are characterised mainly by catalytic versatility and application in different industrial segments. The aim of this study was to biochemically characterise a lipase from a new strain of Bacillus sp. ITP-001. The isoelectric point and molecular mass were 3.12 and 54 kDa, respectively. The optima lipase activity was 276 U g-1 at pH 7.0 and a temperature of 80 ºC, showing greater stability at pH 5.0 and 37 ºC. Enzymatic activity was stimulated by various ions and pyridine, and inhibited by Cu+ and ethanol. The values of Km and v max were 105.26 mmol and 0.116 mmol min-1 g-1, respectively determined by the Eadie-Scatchard method.

  9. Screening of Bacillus coagulans strains in lignin supplemented minimal medium with high throughput turbidity measurements

    Directory of Open Access Journals (Sweden)

    Robert Glaser

    2014-12-01

    Full Text Available The aim of this study was to extend the options for screening and characterization of microorganism through kinetic growth parameters. In order to obtain data, automated turbidimetric measurements were accomplished to observe the response of strains of Bacillus coagulans. For the characterization, it was decided to examine the influence of varying concentrations of lignin with respect to bacterial growth. Different mathematical models are used for comparison: logistic, Gompertz, Baranyi and Richards and Stannard. The growth response was characterized by parameters like maximum growth rate, maximum population, and the lag time. In this short analysis we present a mathematical approach towards a comparison of different microorganisms. Furthermore, it can be demonstrated that lignin in low concentrations can have a positive influence on the growth of B. coagulans.

  10. Isolation of Bacillus strains from the rhizosphere of cereals and in vitro screening for antagonism against phytopathogenic, food-borne pathogenic and spoilage micro-organisms.

    Science.gov (United States)

    Földes, T; Bánhegyi, I; Herpai, Z; Varga, L; Szigeti, J

    2000-11-01

    Bacillus strains were isolated from the rhizosphere of cereals in order to be used as natural biocontrol agents (BCAs). They were screened for antagonism in vitro against various test micro-organisms. The isolates showing antagonism were identified to species level. A combination of techniques was employed for the isolation of Bacillus species. Using the direct method, only one of the 25 isolates screened showed antagonistic properties. This strain (IFS-01) was identified by means of API test strips and the ATB Plus computer programme. It proved to be Bacillus subtilis and consequently has been designated as Bacillus subtilis IFS-01. This strain produced either a broad spectrum antimicrobial compound or several compounds with different activities. The fungi and Gram-positive bacteria were more sensitive to the antagonistic isolate than the Gram-negative bacteria. A Bacillus strain producing BCAs which can be used as biopesticides or organic preservatives has been isolated and identified.

  11. The characteristics of a novel heterotrophic nitrification-aerobic denitrification bacterium, Bacillus methylotrophicus strain L7.

    Science.gov (United States)

    Zhang, Qing-Ling; Liu, Ying; Ai, Guo-Min; Miao, Li-Li; Zheng, Hai-Yan; Liu, Zhi-Pei

    2012-03-01

    Bacillus methylotrophicus strain L7, exhibited efficient heterotrophic nitrification-aerobic denitrification ability, with maximum NH(4)(+)-N and NO(2)(-)-N removal rate of 51.58 mg/L/d and 5.81 mg/L/d, respectively. Strain L7 showed different gaseous emitting patterns from those strains ever described. When (15)NH(4)Cl, or Na(15)NO(2), or K(15)NO(3) was used, results of GC-MS indicated that N(2)O was emitted as the intermediate of heterotrophic nitrification or aerobic denitrification, while GC-IRMS results showed that N(2) was produced as end product when nitrite was used. Single factor experiments suggested that the optimal conditions for heterotrophic nitrification were sodium succinate as carbon source, C/N 6, pH 7-8, 0 g/L NaCl, 37 °C and a wide range of NH(4)(+)-N from 80 to 1000 mg/L. Orthogonal tests showed that the optimal conditions for aerobic denitrification were C/N 20, pH 7-8, 10 g/L NaCl and DO 4.82 mg/L (shaking speed 50 r/min) when nitrite was served as substrate.

  12. Lipopeptides from Bacillus strain AR2 inhibits biofilm formation by Candida albicans.

    Science.gov (United States)

    Rautela, Ria; Singh, Anil Kumar; Shukla, Abha; Cameotra, Swaranjit Singh

    2014-05-01

    The ability of the human fungal pathogen Candida albicans to reversibly switch between different morphological forms and establish biofilms is crucial for establishing infection. Targeting phenotypic plasticity and biofilm formation in C. albicans represents a new concept for antifungal drug discovery. The present study evaluated the influence of cyclic lipopeptide biosurfactant produced by Bacillus amyloliquefaciens strain AR2 on C. albicans biofilms. The biosurfactant was characterized as a mixture of iturin and fengycin by MALDI-TOF and amino acid analysis. The biosurfactant exhibited concentration dependent growth inhibition and fungicidal activity. The biosurfactant at sub-minimum growth inhibition concentration decreased cell surface hydrophobicity, hindered germ tube formation and reduced the mRNA expression of hyphae-specific gene HWP1 and ALS3 without exhibiting significant growth inhibition. The biosurfactants inhibited biofilm formation in the range of 46-100 % depending upon the concentration and Candida strains. The biosurfactant treatment dislodged 25-100 % of preformed biofilm from polystyrene plates. The biosurfactant retained its antifungal and antibiofilm activity even after exposure to extreme temperature. By virtue of the ability to inhibit germ tube and biofilm formation, two important traits of C. albicans involved in establishing infection, lipopeptides from strain AR2 may represent a potential candidate for developing heat stable anti-Candida drugs.

  13. Isolation and characterization of different strains of Bacillus licheniformis for the production of commercially significant enzymes.

    Science.gov (United States)

    Ghani, Maria; Ansari, Asma; Aman, Afsheen; Zohra, Rashida Rahmat; Siddiqui, Nadir Naveed; Qader, Shah Ali Ul

    2013-07-01

    Utilization of highly specific enzymes for various industrial processes and applications has gained huge momentum in the field of white biotechnology. Selection of a strain by efficient plate-screening method for a specific purpose has also favored and boosted the isolation of several industrially feasible microorganisms and screening of a large number of microorganisms is an important step in selecting a potent culture for multipurpose usage. Five new bacterial isolates of Bacillus licheniformis were discovered from indigenous sources and characterized on the basis of phylogeny using 16S rDNA gene analysis. Studies on morphological and physiological characteristics showed that these isolates can easily be cultivated at different temperatures ranging from 30°C to 55°C with a wide pH values from 3.0 to 11.0 All these 05 isolates are salt tolerant and can grow even in the presences of high salt concentration ranging from 7.0 to 12.0%. All these predominant isolates of B. licheniformis strains showed significant capability of producing some of the major industrially important extracellular hydrolytic enzymes including α-amylase, glucoamylase, protease, pectinase and cellulase in varying titers. All these isolates hold great potential as commercial strains when provided with optimum fermentation conditions.

  14. Lactic Acid Production from Pretreated Hydrolysates of Corn Stover by a Newly Developed Bacillus coagulans Strain.

    Directory of Open Access Journals (Sweden)

    Ting Jiang

    Full Text Available An inhibitor-tolerance strain, Bacillus coagulans GKN316, was developed through atmospheric and room temperature plasma (ARTP mutation and evolution experiment in condensed dilute-acid hydrolysate (CDH of corn stover. The fermentabilities of other hydrolysates with B. coagulans GKN316 and the parental strain B. coagulans NL01 were assessed. When using condensed acid-catalyzed steam-exploded hydrolysate (CASEH, condensed acid-catalyzed liquid hot water hydrolysate (CALH and condensed acid-catalyzed sulfite hydrolysate (CASH as substrates, the concentration of lactic acid reached 45.39, 16.83, and 18.71 g/L by B. coagulans GKN316, respectively. But for B. coagulans NL01, only CASEH could be directly fermented to produce 15.47 g/L lactic acid. The individual inhibitory effect of furfural, 5-hydroxymethylfurfural (HMF, vanillin, syringaldehyde and p-hydroxybenzaldehyde (pHBal on xylose utilization by B. coagulans GKN316 was also studied. The strain B. coagulans GKN316 could effectively convert these toxic inhibitors to the less toxic corresponding alcohols in situ. These results suggested that B. coagulans GKN316 was well suited to production of lactic acid from undetoxified lignocellulosic hydrolysates.

  15. Altered protoxin activation by midgut enzymes from a Bacillus thuringiensis resistant strain of Plodia interpunctella.

    Science.gov (United States)

    Oppert, B; Kramer, K J; Johnson, D E; MacIntosh, S C; McGaughey, W H

    1994-02-15

    Processing of Bacillus thuringiensis protoxins to toxins by midgut proteinases from a strain of the Indianmeal moth, Plodia interpunctella (Hubner), resistant to B. thuringiensis subspecies entomocidus (HD-198) was slower than that by midgut proteinases from the susceptible parent strain or a strain resistant to B. thuringiensis subspecies kurstaki (HD-1, Dipel). Midgut extracts from entomocidus-resistant insects exhibited five-fold lower activity toward the synthetic substrate alpha-N-benzoyl-DL-arginine rho-nitroanilide than extracts from susceptible or kurstaki-resistant insects. Midgut enzymes from susceptible or kurstaki-resistant insects converted the 133 kDa CryIA(c) protoxin to 61-63 kDa proteins, while incubations with entomocidus-resistant enzymes resulted in predominantly products of intermediate size, even with increased amounts of midgut extract. The 61-63 kDa proteins were only produced by entomocidus-resistant midgut extracts after long term incubations with the protoxin. The data suggest that altered protoxin activation by midgut proteinases is involved in some types of insect resistance to B. thuringiensis.

  16. Evaluation of genetic and phenotypic consistency of Bacillus coagulans MTCC 5856: a commercial probiotic strain.

    Science.gov (United States)

    Majeed, Muhammed; Nagabhushanam, Kalyanam; Natarajan, Sankaran; Sivakumar, Arumugam; Eshuis-de Ruiter, Talitha; Booij-Veurink, Janine; de Vries, Ynte P; Ali, Furqan

    2016-04-01

    Commercial probiotics preparation containing Bacillus coagulans have been sold in the market for several decades. Due to its high intra-species genomic diversity, it is very likely that B. coagulans strain may alter in different ways over multiple years of production. Therefore, the present study focuses to evaluate the genetic consistency and probiotic potential of B. coagulans MTCC 5856. Phenotypic and genotypic techniques including biochemical profiling, 16S rRNA sequencing, GTG 5″, BOX PCR fingerprinting, and Multi-Locus-Sequence typing (MLST) were carried out to evaluate the identity and consistency of the B. coagulans MTCC 5856. Further, in vitro probiotic potential, safety and stability at ambient temperature conditions of B. coagulans MTCC 5856 were evaluated. All the samples were identified as B. coagulans by biochemical profiling and 16S rRNA sequencing. GTG 5″, BOX PCR fingerprints and MLST studies revealed that the same strain was present over 3 years of commercial production. B. coagulans MTCC 5856 showed resistance to gastric acid, bile salt and exhibited antimicrobial activity in in-vitro studies. Additionally, B. coagulans MTCC 5856 was found to be non-mutagenic, non-cytotoxic, negative for enterotoxin genes and stable at ambient temperature (25 ± 2 °C) for 36 months. The data of the study verified that the same strain of B. coagulans MTCC 5856 was present in commercial preparation over multiple years of production.

  17. Isolation and characterization of a furfural-degrading bacterium Bacillus cereus sp. strain DS1.

    Science.gov (United States)

    Zheng, Dan; Bao, Jianguo; Lu, Jueming; Gao, Chunlei

    2015-02-01

    Furfural was found to be the main organic pollutant in the wastewater coming from the Diosgenin factory. This substance is derived from acidic pentosan in Dioscorea zingiberensis and is also found in a variety of agricultural byproducts, including corncobs, oat, wheat bran, and sawdust. It is regarded as a toxicant and an inhibitor to the growth of microorganism in both sewage disposal and biological fermentation. A furfural-degrading strain (DS1) was isolated from activated sludge of wastewater treatment plant in a diosgenin factory by continuous enrichment culture. The strain was identified as Bacillus cereus based on morphological, physiological tests, as well as on 16S rDNA sequence and Biolog analyses. The capacity of this strain to grow on a mineral salt medium, utilizing furfural as the sole carbon and energy source to degrade furfural, was investigated in this study. Under the condition of pH 9.0, temperature 35 °C, with rotating speed of 150 rpm, and an inoculum of 6 %, the strain showed that the furfural degradation capacity reaches 35 % in 7 days, as measured by high-performance liquid chromatography. The addition of inorganic carbon sources could bring down the biodegradation efficiency of the furfural. The strain DS1 showed better furfural removal capacity, as compared to other inorganic carbon sources in the media. Furthermore, a furfural concentration of as high as 4,000 mg L(-1) was tolerated by the culture. The capacity to degrade furfural was demonstrated for the first time by using the genus B. cereus. This study suggests the possible application in biodegradation strategies.

  18. PCR screening for the surfactin (sfp) gene in marine Bacillus strains and its molecular characterization from Bacillus tequilensis NIOS11

    OpenAIRE

    POROB, Seema; NAYAK, Sagar; FERNANDES, Areena; PADMANABHAN, Priyanka

    2013-01-01

    The sfp gene responsible for surfactin production was screened from the DNA extracts of 37 Bacillus spp. whose identity was confirmed by 16S rRNA gene sequence analyses. PCR screening revealed amplification of sfp gene fragments in a total of 25 isolates. Several isolates belonging to Bacillus tequilensis were found to be positive for this gene. A gene fragment coding for the sfp gene was amplified and cloned from genomic DNA of the isolate B. tequilensis NIOS11. The cloned gene has an open r...

  19. Characterization of cry2-type genes of Bacillus thuringiensis strains from soil-isolated of Sichuan basin, China

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    Hongxia Liang

    2011-03-01

    Full Text Available Sichuan basin, situated in the west of China, is the fourth biggest basin in China. In order to describe a systematic study of the cry2-type genes resources from Bacillus thuringiensis strains of Sichuan basin, a total of 791 Bacillus thuringiensis strains have been screened from 2650 soil samples in different ecological regions. The method of PCR-restriction fragment length polymorphism (PCR-RFLP was used to identify the type of cry2 genes. The results showed that 322 Bacillus thuringiensis strains harbored cry2-type genes and four different RFLP patterns were found. The combination of cry2Aa/cry2Ab genes was the most frequent (90.4%, followed by cry2Aa (6.8% and cry2Ab alone (2.5%, and only one novel type of cry2 gene was cloned from one isolate (JF19-2. The full-length of this novel gene was obtained by the method of thermal asymmetric interlaced PCR (Tail-PCR, which was designated as cry2Ag1 (GenBank No. ACH91610 by the Bt Pesticide Crystal Protein Nomenclature Committee. In addition, the result of scanning electron microscopic (SEM observation showed that these strains had erose, spherical, bipyramidal, and square crystal. And the results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE demonstrated that these strains harbored about one to three major proteins. These strains exhibited a wide range of insecticidal spectrum toxic to Aedes aegypti (Diptera and Pieris rapae Linnaeus, 1758 (Lepidoptera. Particularly, JF19-2 contained cry2Ag gene had the highest insecticidal activity. All these researches mentioned above revealed the diversity and particularity of cry2-type gene resources from Bacillus thuringiensis strains in Sichuan basin.

  20. Deciphering the conserved genetic loci implicated in plant disease control through comparative genomics of Bacillus amyloliquefaciens subsp. plantarum strains

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    Mohammad J Hossain

    2015-08-01

    Full Text Available To understand the growth-promoting and disease-inhibiting activities of plant growth-promoting rhizobacteria (PGPR strains, the genomes of 12 Bacillus subtilis group strains with PGPR activity were sequenced and analyzed. These B. subtilis strains exhibited high genomic diversity, whereas the genomes of B. amyloliquefaciens strains (a member of the B. subtilis group are highly conserved. A pairwise BLASTp matrix revealed that gene family similarity among Bacillus genomes ranges from 32- 90%, with 2,839 genes within the core genome of B. amyloliquefaciens subsp. plantarum. Comparative genomic analyses of B. amyloliquefaciens strains identified genes that are linked with biological control and colonization of roots and/or leaves, including 73 genes uniquely associated with subsp. plantarum strains that have predicted functions related to signaling, transportation, secondary metabolite production, and carbon source utilization. Although B. amyloliquefaciens subsp. plantarum strains contain gene clusters that encode many different secondary metabolites, only polyketide biosynthetic clusters that encode difficidin and macrolactin are conserved within this subspecies. To evaluate their role in plant pathogen biocontrol, genes involved in secondary metabolite biosynthesis were deleted in B. amyloliquefaciens subsp. plantarum strain, revealing that difficidin expression is critical in reducing the severity of disease, caused by Xanthomonas axonopodis pv. vesicatoria in tomato plants. This study defines genomic features of PGPR strains and links them with biocontrol activity and with host colonization.

  1. Bacillus cereus strain isolated from Demodex folliculorum in patients with topical steroid-induced rosaceiform facial dermatitis*

    Science.gov (United States)

    Tatu, Alin Laurentiu; Ionescu, Marius Anton; Clatici, Victor Gabriel; Cristea, Violeta Corina

    2016-01-01

    The aim of the study was to identify Bacillus species from the Demodex folliculorum of patients with topical steroidinduced facial rosaceiform dermatitis. Of the 75 patients examined, 20% had clinical spinulosis, while 18.66% had dermoscopic features of Demodex: follicular plugs and tails. Of the 17.33% positive patients identified upon microscopy for Demodex, samples for bacterial culture were plated on trypticase soy Colombia agar. Identification was performed by microorganisms grown method mass spectrometry. We identified a strain of Bacillus cereus. PMID:27828651

  2. PCR screening for the surfactin (sfp) gene in marine Bacillus strains and its molecular characterization from Bacillus tequilensis NIOS11

    Digital Repository Service at National Institute of Oceanography (India)

    Porob, S.; Nayak, S.; Fernandes, Areena; Padmanabhan, P.; Patil, B.A.; Meena, R.M.; Ramaiah, N.

    The sfp gene responsible for surfactin production was screened from the DNA extracts of 37 Bacillus spp. whose identity was confirmed by 16S rRNA gene sequence analyses. PCR screening revealed amplification of sfp gene fragments in a total of 25...

  3. Calcium Carbonate Precipitation by Bacillus and Sporosarcina Strains Isolated from Concrete and Analysis of the Bacterial Community of Concrete.

    Science.gov (United States)

    Kim, Hyun Jung; Eom, Hyo Jung; Park, Chulwoo; Jung, Jaejoon; Shin, Bora; Kim, Wook; Chung, Namhyun; Choi, In-Geol; Park, Woojun

    2016-03-01

    Microbially induced calcium carbonate precipitation (CCP) is a long-standing but re-emerging environmental engineering process for production of self-healing concrete, bioremediation, and long-term storage of CO2. CCP-capable bacteria, two Bacillus strains (JH3 and JH7) and one Sporosarcina strain (HYO08), were isolated from two samples of concrete and characterized phylogenetically. Calcium carbonate crystals precipitated by the three strains were morphologically distinct according to field emission scanning electron microscopy. Energy dispersive X-ray spectrometry mapping confirmed biomineralization via extracellular calcium carbonate production. The three strains differed in their physiological characteristics: growth at alkali pH and high NaCl concentrations, and urease activity. Sporosarcina sp. HYO08 and Bacillus sp. JH7 were more alkali- and halotolerant, respectively. Analysis of the community from the same concrete samples using barcoded pyrosequencing revealed that the relative abundance of Bacillus and Sporosarcina species was low, which indicated low culturability of other dominant bacteria. This study suggests that calcium carbonate crystals with different properties can be produced by various CCP-capable strains, and other novel isolates await discovery.

  4. Survival and retention of the probiotic properties of Bacillus sp. strains under marine stress starvation conditions and their potential use as a probiotic in Artemia culture.

    Science.gov (United States)

    Mahdhi, Abdelkarim; Esteban, Maria Ángeles; Hmila, Zeineb; Bekir, Karima; Kamoun, Fathi; Bakhrouf, Amina; Krifi, Boubaker

    2012-12-01

    The probiotic properties of Bacillus strains isolated from Artemia culture and the effect of marine stress on viability and survival were investigated, as well as the changes occurring in their properties. Analyses showed that these bacteria corresponded to the genus Bacillus sp. Antagonism and adherence assays revealed that Bacillus strains have an inhibitory effect against tested pathogenic bacteria and are fairly adherent. Normal and starved cells showed different enzymatic profiles. Challenge tests performed with Artemia larvae provided evidence that the tested Bacillus strains were neither pathogenic nor toxic to the host and conferred protection for Artemia culture against pathogens. The tested strains maintained their viability and their probiotic properties during the period of study. The results suggest that the tested strains have suffered changes allowing them to survive in seawater in the absence of nutrients and outside their natural host, identifying them as potential probiotic candidates for Artemia culture.

  5. Proteomic Analysis of Bacillus thuringiensis Strain 4.0718 at Different Growth Phases

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    Xiaohui Li

    2012-01-01

    Full Text Available The growth process of Bacillus thuringiensis Bt4.0718 strain was studied using proteomic technologies. The proteins of Bt whole cells at three phases—middle vegetative, early sporulation, and late sporulation—were extracted with lysis buffer, followed with separation by 2-DE and identified by MALDI-TOF/TOF MS. Bioactive factors such as insecticidal crystal proteins (ICPs including Cry1Ac(3, Cry2Aa, and BTRX28, immune inhibitor (InhA, and InhA precursor were identified. InhA started to express at the middle vegetative phase, suggesting its contribution to the survival of Bt in the host body. At the early sporulation phase, ICPs started their expression. CotJC, OppA, ORF1, and SpoIVA related to the formation of crystals and spores were identified, the expression characteristics of which ensured the stable formation of crystals and spores. This study provides an important foundation for further exploration of the stable expression of ICPs, the smooth formation of crystals, and the construction of recombinant strains.

  6. Division of labour and terminal differentiation in a novel Bacillus thuringiensis strain.

    Science.gov (United States)

    Deng, Chao; Slamti, Leyla; Raymond, Ben; Liu, Guiming; Lemy, Christelle; Gominet, Myriam; Yang, Jingni; Wang, Hengliang; Peng, Qi; Zhang, Jie; Lereclus, Didier; Song, Fuping

    2015-02-01

    A major challenge in bacterial developmental biology has been to understand the mechanisms underlying cell fate decisions. Some differentiated cell types display cooperative behaviour. Cooperation is one of the greatest mysteries of evolutionary biology and microbes have been considered as an excellent system for experimentally testing evolution theories. Bacillus thuringiensis (Bt) is a spore-forming bacterium, which is genetically closely related to B. anthracis, the agent of anthrax, and to B. cereus, an opportunistic human pathogen. The defining feature that distinguishes Bt from its relatives is its ability to produce crystal inclusions in the sporulating cells. These toxins are solubilized after ingestion and are cooperative public goods in insect hosts. In this study, we describe a Bt strain LM1212 that presents the unique ability to terminally differentiate into crystal producers and spore formers. Transcriptional analysis based on lacZ and gfp reporter genes suggested that this phenotype is the consequence of a new type of cell differentiation associated with a novel regulation mode of cry gene expression. The differentiating crystal-producer phenotype has higher spore productivity than a typical Bt strain and is better able to compete with Cry toxin null 'cheaters'. Potentially, this division of labour provides additional fitness benefits in terms of spore viability or durability of Cry toxin.

  7. Thiamethoxam degradation by Pseudomonas and Bacillus strains isolated from agricultural soils.

    Science.gov (United States)

    Rana, Shivnam; Jindal, Vikas; Mandal, Kousik; Kaur, Gurpreet; Gupta, V K

    2015-05-01

    Twelve bacterial species were evaluated to know the degradation pattern of thiamethoxam in liquid medium. All the bacterial species could actively degrade phorate in a mineral salt medium containing phorate (50 μg ml(-1)) as sole carbon source. As these species have ability to degrade, we used these for the degradation of thiamethoxam--a neonicoitinoids. Screening of 12 active phorate-metabolizing bacterial species resulted in selection of Bacillus aeromonas strain IMBL 4.1 and Pseudomonas putida strain IMBL 5.2 causing 45.28 and 38.23 % thiamethoxam (50 μg ml(-1)) reduction, respectively, in 15 days as potential thiamethoxam degrading species. These two bacterial species grew optimally at 37 °C under shake culture conditions in MSMT medium raised with initial pH of 6.0-6.5 and use of these optimum cultural conditions resulted in improved thiamethoxam degradation by these bacterial species. These species caused maximum thiamethoxam degradation only in the presence of thiamethoxam as sole source of carbon and energy and the same was reduced in the presence of easily metabolize able carbon (C₀ and C₁) and nitrogen ((N₀, N₁ and N₂) sources. This could be attributed to involvement of repressible metabolic pathways, reactions of which are inhibited by the presence of easily available nutrients for growth. Besides above, qualitative analysis of thiamethoxam residues by gas liquid chromatography revealed complete metabolization of thiamethoxam without detectable accumulation of any known thiamethoxam metabolites.

  8. Beta-glucosidase enzymatic activity of crystal polypeptide of the Bacillus thuringiensis strain 1.1.

    Science.gov (United States)

    Papalazaridou, A; Charitidou, L; Sivropoulou, A

    2003-01-01

    The crystals of Bacillus thuringiensis strain 1.1 consist of the 140 kDa delta-endotoxin, which exhibits beta-glucosidase enzymatic activity, based on the following data. (i) Purified crystals exhibit beta-glucosidase enzymatic activity. When the crystals are reacted with specific antibodies directed either against the commercial (almond purified) beta-glucosidase or against the 140 kDa polypeptide, then considerable reduction of enzymatic activity is observed almost at the same level with both antibodies. (ii) Commercial beta-glucosidase and the 140 kDa crystal polypeptide share antigenic similarities; in Western immunoblots, the 140 kDa crystal polypeptide is recognized by anti-beta-glucosidase antibodies, and commercial beta-glucosidase is recognized by anti-140-kDa antibodies. (iii) The enzymatic properties of commercial beta-glucosidase and that resident in the crystals of B. thuringiensis strain 1.1 are very similar. Thus, both enzymes hydrolyze a wide range of substrates (aryl-beta-glucosides, disaccharides with alpha- or beta-linkage polysaccharides) and have an optimum activity at 40 degrees C and pH 5. Both enzymes are relatively thermostable and are resistant to end-product inhibition by glucose. Additionally, they show the same pattern of inhibition or activation by several chemical compounds. (iv) The crystals and commercial beta-glucosidase show almost equivalent levels of insecticidal activity against Drosophila melanogaster larvae and, furthermore, cause reduction in adult flies that emerge from larvae surviving treatment.

  9. Biosorption of cationic basic dye and cadmium by the novel biosorbent Bacillus catenulatus JB-022 strain.

    Science.gov (United States)

    Kim, Su Young; Jin, Mi Ra; Chung, Chang Ho; Yun, Yeoung-Sang; Jahng, Kwang Yeop; Yu, Kang-Yeol

    2015-04-01

    Biosorption of heavy metals and dyes is a promising technology that involves the removal of toxic metals from industrial wastes. The present study aims to screen the bacterial strains isolated from soils and polluted pond for their potential biosorption of both cationic dye and cadmium. Bacillus catenulatus JB-022 strain removed 58% and 66% of cationic basic blue 3 (BB3) and cadmium (Cd(II)) at the respective concentrations of 2000 mg/L and 150 mg/L. The biosorption equilibrium data were well fitted by the Langmuir adsorption isotherm, and the kinetic studies indicated that the biosorption followed the pseudo-second-order model. The biosorption kinetics showed that the equilibrium was reached within 10 min and 5 min for BB3 and Cd(II), respectively. According to the Langmuir model, the maximum uptakes of BB3 and Cd(II) by the JB-022 biomass were estimated to be 139.74 and 64.28 mg/g, respectively. To confirm the surface morphology and functional groups, field emission scanning electron microscope, energy-dispersive X-ray spectrometer, X-ray diffraction, and Fourier transform infrared spectroscopy analyses were carried out, and the results revealed that the biomass of JB-022 has carboxyl and phosphonate groups as potential surface functional groups capable of binding to cationic pollutants. In conclusion, B. catenulatus JB-022 is proposed as an excellent biosorbent with potentially important applications in removal of cationic pollutants from wastewaters.

  10. Antagonistic action of Bacillus subtilis strain SG6 on Fusarium graminearum.

    Science.gov (United States)

    Zhao, Yueju; Selvaraj, Jonathan Nimal; Xing, Fuguo; Zhou, Lu; Wang, Yan; Song, Huimin; Tan, Xinxin; Sun, Lichao; Sangare, Lancine; Folly, Yawa Minnie Elodie; Liu, Yang

    2014-01-01

    Fusarium graminearum causes Fusarium head blight (FHB), a devastating disease that leads to extensive yield and quality loss of wheat and barley. Bacteria isolated from wheat kernels and plant anthers were screened for antagonistic activity against F. graminearum. Based on its in vitro effectiveness, strain SG6 was selected for characterization and identified as Bacillus subtilis. B. subtilis SG6 exhibited a high antifungal effect on the mycelium growth, sporulation and DON production of F. graminearum with the inhibition rate of 87.9%, 95.6% and 100%, respectively. In order to gain insight into biological control effect in situ, we applied B. subtilis SG6 at anthesis through the soft dough stage of kernel development in field test. It was revealed that B. subtilis SG6 significantly reduced disease incidence (DI), FHB index and DON (P ≤ 0.05). Further, ultrastructural examination shows that B. subtilis SG6 strain induced stripping of F. graminearum hyphal surface by destroying the cellular structure. When hypha cell wall was damaged, the organelles and cytoplasm inside cell would exude, leading to cell death. The antifungal activity of SG6 could be associated with the coproduction of chitinase, fengycins and surfactins.

  11. Detection of toxigenic Bacillus cereus strains isolated from vegetables in Mexico City.

    Science.gov (United States)

    Flores-Urbán, Karen A; Natividad-Bonifacio, Iván; Vázquez-Quiñones, Carlos R; Vázquez-Salinas, Carlos; Quiñones-Ramírez, Elsa Irma

    2014-12-01

    Bacillus cereus can cause diarrhea and emetic syndromes after ingestion of food contaminated with it. This ability is due to the production of enterotoxins by this microorganism, these being the hemolysin BL complex, which is involved in the diarrheal syndrome, and cereulide, which is responsible for the emetic syndrome. The detection of genes associated with the production of these toxins can predict the virulence of strains isolated from contaminated food. In this paper, we analyzed 100 samples of vegetables, 25 of each kind (broccoli, coriander, carrot, and lettuce) obtained from different markets in Mexico City and its metropolitan area. B. cereus was isolated in 32, 44, 84, and 68% of the samples of broccoli, carrot, lettuce, and coriander, respectively. The hblA gene (encoding one of the three subunits of hemolysin BL) was amplified in 100% of the B. cereus isolates, and the ces gene (encoding the cereulide) could not be amplified from any of them. This is the first report of B. cereus isolation from the vegetables analyzed in this work and, also, the first report in Mexico of the isolation from vegetables of strains with potential virulence. The results should serve as evidence of the potential risk of consuming these foods without proper treatment.

  12. Purification and characterization of an acidothermophilic cellulase enzyme produced by Bacillus subtilis strain LFS3.

    Science.gov (United States)

    Rawat, Rekha; Tewari, Lakshmi

    2012-07-01

    In the present investigation, a microorganism hydrolyzing carboxymethylcellulose (CMC) was isolated and identified as Bacillus subtilis strain LFS3 by 16S rDNA sequence analysis. The carboxymethylcellulase (CMCase) enzyme produced by the B. subtilis strain LFS3 was purified by (NH₄)₂SO₄ precipitation, ion exchange and gel filtration chromatography, with an overall recovery of 15 %. Native-PAGE analysis of purified CMCase revealed the molecular weight of enzyme to be about 185 kDa. The activity profile of CMCase enzyme showed the optimum activity at temperature 60 °C and pH 4.0, respectively. The enzyme activity was induced by Na⁺, Mg²⁺, NH₄⁺, and EDTA, whereas strongly inhibited by Hg²⁺ and Fe³⁺. The purified enzyme hydrolyzed CMC, filter paper, and xylan, but not p-nitrophenyl β-D-glucopyranoside and cellulose. Kinetic analysis of purified enzyme showed the K(m) value of 2.2 mg/ml. Thus, acidophilic as well as thermophilic nature makes this cellulase a suitable candidate for current mainstream biomass conversion into fuel and other industrial processes.

  13. Bacillus cereus strain S2 shows high nematicidal activity against Meloidogyne incognita by producing sphingosine

    Science.gov (United States)

    Gao, Huijuan; Qi, Gaofu; Yin, Rong; Zhang, Hongchun; Li, Chenggang; Zhao, Xiuyun

    2016-01-01

    Plant-parasitic nematodes cause serious crop losses worldwidely. This study intended to discover the antagonistic mechanism of Bacillus cereus strain S2 against Meloidogyne incognita. Treatment with B. cereus strain S2 resulted in a mortality of 77.89% to Caenorhabditis elegans (a model organism) and 90.96% to M. incognita. In pot experiment, control efficiency of B. cereus S2 culture or supernatants were 81.36% and 67.42% towards M. incognita, respectively. In field experiment, control efficiency was 58.97% towards M. incognita. Nematicidal substances were isolated from culture supernatant of B. cereus S2 by polarity gradient extraction, silica gel column chromatography and HPLC. Two nematicidal compounds were identified as C16 sphingosine and phytosphingosine by LC-MS. The median lethal concentration of sphingosine was determined as 0.64 μg/ml. Sphingosine could obviously inhibit reproduction of C. elegans, with an inhibition rate of 42.72% for 24 h. After treatment with sphingosine, ROS was induced in intestinal tract, and genital area disappeared in nematode. Furthermore, B. cereus S2 could induce systemic resistance in tomato, and enhance activity of defense-related enzymes for biocontrol of M. incognita. This study demonstrates the nematicidal activity of B. cereus and its product sphingosine, as well provides a possibility for biocontrol of M. incognita. PMID:27338781

  14. Antagonistic action of Bacillus subtilis strain SG6 on Fusarium graminearum.

    Directory of Open Access Journals (Sweden)

    Yueju Zhao

    Full Text Available Fusarium graminearum causes Fusarium head blight (FHB, a devastating disease that leads to extensive yield and quality loss of wheat and barley. Bacteria isolated from wheat kernels and plant anthers were screened for antagonistic activity against F. graminearum. Based on its in vitro effectiveness, strain SG6 was selected for characterization and identified as Bacillus subtilis. B. subtilis SG6 exhibited a high antifungal effect on the mycelium growth, sporulation and DON production of F. graminearum with the inhibition rate of 87.9%, 95.6% and 100%, respectively. In order to gain insight into biological control effect in situ, we applied B. subtilis SG6 at anthesis through the soft dough stage of kernel development in field test. It was revealed that B. subtilis SG6 significantly reduced disease incidence (DI, FHB index and DON (P ≤ 0.05. Further, ultrastructural examination shows that B. subtilis SG6 strain induced stripping of F. graminearum hyphal surface by destroying the cellular structure. When hypha cell wall was damaged, the organelles and cytoplasm inside cell would exude, leading to cell death. The antifungal activity of SG6 could be associated with the coproduction of chitinase, fengycins and surfactins.

  15. Solid state bioreactor production of transglutaminase by Amazonian Bacillus circulans BL32 strain.

    Science.gov (United States)

    de Souza, Claucia Fernanda Volken; Heck, Júlio Xandro; Ayub, Marco Antônio Záchia

    2008-12-01

    In this work, we investigated the production of transglutaminase (TGase) by an Amazonian isolated strain of Bacillus circulans by solid-state cultivation (SSC). Several agro-industrial residues, such as untreated corn grits, milled brewers rice, industrial fibrous soy residue, soy hull, and malt bagasse, were used as substrates for microbial growth and enzyme production. Growth on industrial fibrous soy residue, which is rich in protein and hemicellulose, produced the highest TGase activity (0.74 U g(-1) of dried substrate after 48 h of incubation). A 2(3) central composite design was applied to determine the optimal conditions of aeration, cultivation temperature and inoculum cell concentration to TGase production. The best culture conditions were determined as being 0.6 L air min(-1), 33 degrees C and 10 log (10) CFU g(-1) of dried substrate, respectively. Under the proposed optimized conditions, the model predicted an enzyme production of 1.16 U g(-1) of dried substrate, closely matching the experimental activity of 1.25 U g(-1). Results presented in this work point to the use of this newly isolated B. circulans strain as a potential alternative of microbial source for TGase production by SSC, using inexpensive culture media.

  16. New bacteriocin from Bacillus clausii strainGM17: purification, characterization, and biological activity.

    Science.gov (United States)

    Mouloud, Ghadbane; Daoud, Harzallah; Bassem, Jaouadi; Laribi Atef, Ibn; Hani, Belhadj

    2013-12-01

    A bacteriocin-producing strain (9,000 AU/ml) was isolated from the rhizosphere of Algerian healthy plants Ononis angustissima Lam. and identified as Bacillus clausii strain GM17. The bacteriocin, called Bac-GM17, was purified from the culture supernatant after heat treatment, ammonium sulfate precipitation, Sephadex G-50 chromatography and Mono Q fast-performance liquid chromatography (FPLC). Based on matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis, the purified Bac-GM17 is a monomer protein with a molecular mass of 5,158.11 Da. The N-terminal sequencing allowed for the straightforward identification of its first 20 residues, which were of pure bacteriocin. It also revealed that this bacteriocin contained a unique sequence, namely DWTCSKWSCLVCDDCSVELT, which suggests the identification of a novel compound. Bac-GM17 was extremely heat stable (20 min at 120 °C) and was stable within the pH range (3-9). It was found to be resistant to the proteolytic action of trypsin, pepsin, papain, pronase E, and proteinase K. It was also noted to display a bactericidal mode of action against Agrobacterium tumefaciens C58 and a fungistatic mode of action against Candida tropicalis R2 CIP203.

  17. Production and characterization of PHB from two novel strains of Bacillus spp. isolated from soil and activated sludge.

    Science.gov (United States)

    Thirumala, M; Reddy, Sultanpuram Vishnuvardhan; Mahmood, S K

    2010-03-01

    The present study reports two bacteria, designated 87I and 112A, which were isolated from soil and activated sludge samples from Hyderabad, India, and that are capable of producing poly-3-hydroxybutyrate (PHB). Based on phenotypical features and genotypic investigations, these microorganisms were identified as Bacillus spp. Their optimal growth occurred between 28 degrees C and 30 degrees C and pH 7. Bacillus sp. 87I yielded a maximum of 70.04% dry cell weight (DCW) PHB in medium containing glucose as carbon source, followed by 55.5% DCW PHB in lactose-containing medium, whereas Bacillus sp. 112A produced a maximum of 67.73% PHB from glucose, 58.5% PHB from sucrose, followed by 50.5% PHB from starch as carbon substrates. The viscosity average molecular mass (M (v)) of the polymers from Bacillus sp. 87I was 513 kDa and from Bacillus sp. 112A was 521 kDa. All the properties of the biopolymers produced by the two strains 87I and 112A were characterized.

  18. Whole-Genome Sequence and Fosfomycin Resistance of Bacillus sp. Strain G3(2015) Isolated from Seawater off the Coast of Malaysia

    Science.gov (United States)

    Chan, Xin-Yue; Chen, Jian-Woon; Adrian, Tan-Guan-Sheng; Hong, Kar-Wai; Chang, Chien-Yi; Yin, Wai-Fong

    2017-01-01

    ABSTRACT Bacillus sp. is a Gram-positive bacterium that is commonly found in seawater. In this study, the genome of marine Bacillus sp. strain G3(2015) was sequenced using MiSeq. The fosfomycin resistant gene fosB was identified upon bacterial genome annotation. PMID:28360153

  19. Draft Genome Sequence of the Nonpathogenic, Thermotolerant, and Exopolysaccharide-Producing Bacillus anthracis Strain PFAB2 from Panifala Hot Water Spring in West Bengal, India

    Science.gov (United States)

    Banerjee, Aparna; Halder, Urmi; Chaudhry, Vasvi; Varshney, Rajeev K.; Mantri, Shrikant

    2016-01-01

    Bacillus anthracis is the causative agent of fatal anthrax in both animals and humans. It is prevalently pathogenic. Here, we present a Bacillus anthracis PFAB2 strain from a relatively unexplored Panifala hot water spring in West Bengal, India. It is nonpathogenic, exopolysaccharide producing, and thermotolerant in nature. PMID:28007848

  20. Draft Genome Sequence of the Obligate Halophilic Bacillus sp. Strain NSP22.2, Isolated from a Seasonal Salt Marsh of the Great Rann of Kutch, India

    Science.gov (United States)

    Pal, Kamal Krishna; Sherathia, Dharmesh; Vanpariya, Sejal; Patel, Ilaxi; Dalsania, Trupti; Savsani, Kinjal; Sukhadiya, Bhoomika; Mandaliya, Mona; Thomas, Manesh; Ghorai, Sucheta; Rupapara, Rupal; Rawal, Priya

    2013-01-01

    Here, we report the 4.0-Mbp draft genome of an obligate halophile, Bacillus sp. strain NSP22.2, isolated from a seasonal salt marsh of the Great Rann of Kutch, India. To understand the mechanism(s) of obligate halophilism and to isolate the relevant gene(s), the genome of Bacillus sp. NSP22.2 was sequenced. PMID:24356848

  1. Isolation and distribution of mosquito-larvicidal cry genes in Bacillus thuringiensis strains native to Saudi Arabia.

    Science.gov (United States)

    El-kersh, T A; Al-akeel, R A; Al-sheikh, Y A; Alharbi, S A

    2014-12-01

    A total of 157 environmental samples were collected from 11 ecological regions across Saudi Arabia to isolate native Bacillus thuringiensis (Bt) strains. Bt isolates (n=103) were recovered by the 50% (v/v) ethanol treatment method with Bt index range of 0.01 to 0.4. Most of Bt isolates showed spherical crystals (54%), while, irregular, bi-pyramidal, and spore-attached crystal constituted 27, 16 and 3% respectively. PCR analysis with eight general and specific dipteran primers of Cry and Cyt genes, revealed positive amplification for cry4 & cyt1, and cry4A, cry4B and cyt2, and cry 10 and cry 11 genes in 28%, 26%, 22%, and 25% of tested strains respectively; whereas cry2 gene was not detected except with the reference Bt kurstaki HD-1 strain. Bioassays against Aedes caspuis and Culex pipiens larvae indicated that 11 strains displayed better larvicidal activity compared with Bacillus thuringiensis H14 (Bti) reference (LC50 0.6 μg/ml) strain against Ae. caspuis, but only two strains (620A & 633R1, LC50 of 0.09 μg/ml & 0.064 μg/ml) that gave significant enhancement. Additionally, one strain (633R1) showed LC50 similar to that of Bti H14 (LC50 0.064 μg/ml) against Cx. pipiens. With the exception of cyt primers, sequenced DNA of all positive primers amplicons revealed 95 to 99% identity in GenBank with Bacillus thuringiensis subsp. israelensis plasmid pBtoxis and also correlated with its SDS-PAGE expressed protein profiles analysis. It is hoped that our wild bio-insecticide Bt strains can be explored in future in the control of mosquito-vector borne diseases in Saudi Arabia.

  2. Comparative transcriptional profiling of Bacillus cereus sensu lato strains during growth in CO2-bicarbonate and aerobic atmospheres.

    Directory of Open Access Journals (Sweden)

    Karla D Passalacqua

    Full Text Available BACKGROUND: Bacillus species are spore-forming bacteria that are ubiquitous in the environment and display a range of virulent and avirulent phenotypes. This range is particularly evident in the Bacillus cereus sensu lato group; where closely related strains cause anthrax, food-borne illnesses, and pneumonia, but can also be non-pathogenic. Although much of this phenotypic range can be attributed to the presence or absence of a few key virulence factors, there are other virulence-associated loci that are conserved throughout the B. cereus group, and we hypothesized that these genes may be regulated differently in pathogenic and non-pathogenic strains. METHODOLOGY/PRINCIPAL FINDINGS: Here we report transcriptional profiles of three closely related but phenotypically unique members of the Bacillus cereus group--a pneumonia-causing B. cereus strain (G9241, an attenuated strain of B. anthracis (Sterne 34F(2, and an avirulent B. cereus strain (10987--during exponential growth in two distinct atmospheric environments: 14% CO(2/bicarbonate and ambient air. We show that the disease-causing Bacillus strains undergo more distinctive transcriptional changes between the two environments, and that the expression of plasmid-encoded virulence genes was increased exclusively in the CO(2 environment. We observed a core of conserved metabolic genes that were differentially expressed in all three strains in both conditions. Additionally, the expression profiles of putative virulence genes in G9241 suggest that this strain, unlike Bacillus anthracis, may regulate gene expression with both PlcR and AtxA transcriptional regulators, each acting in a different environment. CONCLUSIONS/SIGNIFICANCE: We have shown that homologous and even identical genes within the genomes of three closely related members of the B. cereus sensu lato group are in some instances regulated very differently, and that these differences can have important implications for virulence. This study

  3. Antagonist effects of Bacillus spp. strains against Fusarium graminearum for protection of durum wheat (Triticum turgidum L. subsp. durum).

    Science.gov (United States)

    Zalila-Kolsi, Imen; Ben Mahmoud, Afif; Ali, Hacina; Sellami, Sameh; Nasfi, Zina; Tounsi, Slim; Jamoussi, Kaïs

    2016-11-01

    Bacillus species are attractive due to their potential use in the biological control of fungal diseases. Bacillus amyloliquefaciens strain BLB369, Bacillus subtilis strain BLB277, and Paenibacillus polymyxa strain BLB267 were isolated and identified using biochemical and molecular (16S rDNA, gyrA, and rpoB) approaches. They could produce, respectively, (iturin and surfactin), (surfactin and fengycin), and (fusaricidin and polymyxin) exhibiting broad spectrum against several phytopathogenic fungi. In vivo examination of wheat seed germination, plant height, phenolic compounds, chlorophyll, and carotenoid contents proved the efficiency of the bacterial cells and the secreted antagonist activities to protect Tunisian durum wheat (Triticum turgidum L. subsp. durum) cultivar Om Rabiia against F. graminearum fungus. Application of single bacterial culture medium, particularly that of B. amyloliquefaciens, showed better protection than combinations of various culture media. The tertiary combination of B. amyloliquefaciens, B. subtilis, and P. polymyxa bacterial cells led to the highest protection rate which could be due to strains synergistic or complementary effects. Hence, combination of compatible biocontrol agents could be a strategic approach to control plant diseases.

  4. Anti-biofilm activity of an exopolysaccharide from a sponge-associated strain of Bacillus licheniformis

    Directory of Open Access Journals (Sweden)

    Cordone Angela

    2011-09-01

    Full Text Available Abstract Background Secondary metabolites ranging from furanone to exo-polysaccharides have been suggested to have anti-biofilm activity in various recent studies. Among these, Escherichia coli group II capsular polysaccharides were shown to inhibit biofilm formation of a wide range of organisms and more recently marine Vibrio sp. were found to secrete complex exopolysaccharides having the potential for broad-spectrum biofilm inhibition and disruption. Results In this study we report that a newly identified ca. 1800 kDa polysaccharide having simple monomeric units of α-D-galactopyranosyl-(1→2-glycerol-phosphate exerts an anti-biofilm activity against a number of both pathogenic and non-pathogenic strains without bactericidal effects. This polysaccharide was extracted from a Bacillus licheniformis strain associated with the marine organism Spongia officinalis. The mechanism of action of this compound is most likely independent from quorum sensing, as its structure is unrelated to any of the so far known quorum sensing molecules. In our experiments we also found that treatment of abiotic surfaces with our polysaccharide reduced the initial adhesion and biofilm development of strains such as Escherichia coli PHL628 and Pseudomonas fluorescens. Conclusion The polysaccharide isolated from sponge-associated B. licheniformis has several features that provide a tool for better exploration of novel anti-biofilm compounds. Inhibiting biofilm formation of a wide range of bacteria without affecting their growth appears to represent a special feature of the polysaccharide described in this report. Further research on such surface-active compounds might help developing new classes of anti-biofilm molecules with broad spectrum activity and more in general will allow exploring of new functions for bacterial polysaccharides in the environment.

  5. Production and characterization of a group of bioemulsifiers from the marine Bacillus velezensis strain H3.

    Science.gov (United States)

    Liu, Xiangyang; Ren, Biao; Chen, Ming; Wang, Haibin; Kokare, Chandrakant R; Zhou, Xianlong; Wang, Jidong; Dai, Huanqin; Song, Fuhang; Liu, Mei; Wang, Jian; Wang, Shujin; Zhang, Lixin

    2010-08-01

    Marine microbes are a rich source of bioactive compounds, such as drugs, enzymes, and biosurfactants. To explore the bioactive compounds from our marine natural product library, an oil emulsification assay was applied to discover biosurfactants and bioemulsifiers. A spore-forming bacterial strain from sea mud was found to produce bioemulsifiers with good biosurfactant activity and a broad spectrum of antimicrobial properties. It was identified as Bacillus velezensis H3 using genomic and phenotypic data analysis. This strain was able to produce biosurfactants with an optimum emulsification activity at pH 6.0 and 2% NaCl by using starch as the carbon source and ammonium sulfate as the nitrogen source. The emulsification-guided isolation and purification procedure led to the discovery of the biosurfactant components, which were mainly composed of nC(14)-surfactin and anteisoC(15)-surfactin as determined by NMR and MS spectra. These compounds can reduce the surface tension of phosphate-buffered saline (PBS) from 71.8 to 24.8 mN/m. The critical micelle concentrations (CMCs) of C(14)-surfactin and C(15)-surfactin in 0.1 M PBS (pH 8.0) were determined to be 3.06 x 10(-5) and 2.03 x 10(-5) mol/L, respectively. The surface tension values at CMCs for C(14)-surfactin and C(15)-surfactin were 25.7 and 27.0 mM/m, respectively. In addition, the H3 biosurfactant exhibited antimicrobial activities against Staphyloccocus aureus, Mycobacterium, Klebsiella peneumoniae, Pseudomonas aeruginosa, and Candida albicans. Thus B. velezensis H3 is an alternative surfactin producer with potential application as an industrial strain for the lipopeptide production.

  6. Isolation and characterization of a novel native Bacillus thuringiensis strain BRC-HZM2 capable of degrading chlorpyrifos.

    Science.gov (United States)

    Wu, Songqing; Peng, Yan; Huang, Zhangmin; Huang, Zhipeng; Xu, Lei; Ivan, Gelbič; Guan, Xiong; Zhang, Lingling; Zou, Shuangquan

    2015-03-01

    Studies were carried out to isolate chlorpyrifos degrading Bacillus thuringiensis (Bt) strains from chlorpyrifos-contaminated samples. Six Bt strains (isolation rate 2.7%) were isolated by modified sodium acetate antibiotic heat treatment, and one novel strain (BRC-HZM2) was selected for further analysis. Phenotype and phylogeny analysis of this strain was conducted on the basis of biochemical reactions, antibiotic sensitivity, 16s rRNA genes, plasmid profile, insecticidal crystal protein profiles, and PCR-RFLP for cry and cyt genes. The degradation rate of chlorpyrifos in liquid culture was estimated during 48 h of incubation for the isolate BRC-HZM2. More than 50% of the initial chlorpyrifos concentration degraded within 12 h, 88.9% after 48 h. These results highlight the potential of the Bt strain for biological control and the bioremediation of environments contaminated with chlorpyrifos.

  7. Partial Characterization of an Anti-Candida albicans Bacteriocin Produced by a Marine Strain of Bacillus sp., Sh10

    OpenAIRE

    Fatemeh Shayesteh; Asmat Ahmad; Gires Usup

    2015-01-01

    The bacteriocin-producing strain Bacillus sp., Sh10, isolated from the marine environment, exhibited a broad spectrum of antimicrobial activity against different food spoilage and human pathogens, with a maximum inhibitory activity against Candida albicans. The inhibitory compound was sensitive to trypsin but resistant to proteinase K, lysozyme, lipase and &alpha-amylase. It was heat-stable and remained its activity after autoclaving. In addition, the antimicrobial substance demonstrated stri...

  8. Complete genome sequence of Bacillus thuringiensis CTC-A typical strain with high production of S-layer proteins.

    Science.gov (United States)

    Dong, Zhaoxia; Li, Junhua; Zheng, Jinshui; Geng, Ce; Peng, Donghai; Sun, Ming

    2016-02-20

    Bacillus thuringiensis CTC, which is identified as serotype H2, serovar. finitimus, is high production of S-layer protein. Due to the property of forming isoporous lattices on the whole cell surface, S-layer protein has been widely used in (nano) biotechnology, biomimetics, biomedicine, especially been employed for displaying many important active proteins. Here, we report the complete genome of strain CTC, which contains one circular chromosome and one linear plasmid.

  9. An in-depth characterization of the entomopathogenic strain Bacillus pumilus 15.1 reveals that it produces inclusion bodies similar to the parasporal crystals of Bacillus thuringiensis.

    Science.gov (United States)

    Garcia-Ramon, Diana C; Molina, C Alfonso; Osuna, Antonio; Vílchez, Susana

    2016-04-01

    In the present work, the local isolate Bacillus pumilus 15.1 has been morphologically and biochemically characterized in order to gain a better understanding of this novel entomopathogenic strain active against Ceratitis capitata. This strain could represent an interesting biothechnological tool for the control of this pest. Here, we report on its nutrient preferences, extracellular enzyme production, motility mechanism, biofilm production, antibiotic suceptibility, natural resistance to chemical and physical insults, and morphology of the vegetative cells and spores. The pathogen was found to be β-hemolytic and susceptible to penicillin, ampicillin, chloramphenicol, gentamicin, kanamycin, rifampicin, tetracycline, and streptomycin. We also report a series of biocide, thermal, and UV treatments that reduce the viability of B. pumilus 15.1 by several orders of magnitude. Heat and chemical treatments kill at least 99.9 % of vegetative cells, but spores were much more resistant. Bleach was the only chemical that was able to completely eliminate B. pumilus 15.1 spores. Compared to the B. subtilis 168 spores, B. pumilus 15.1 spores were between 2.67 and 350 times more resistant to UV radiation while the vegetative cells of B. pumilus 15.1 were almost up to 3 orders of magnitude more resistant than the model strain. We performed electron microscopy for morphological characterization, and we observed geometric structures resembling the parasporal crystal inclusions synthesized by Bacillus thuringiensis. Some of the results obtained here such as the parasporal inclusion bodies produced by B. pumilus 15.1 could potentially represent virulence factors of this novel and potentially interesting strain.

  10. Deletion analysis of the C-terminal region of the alpha-amylase of Bacillus sp. strain TS-23.

    Science.gov (United States)

    Lo, Huei-Fen; Lin, Long-Liu; Chiang, Wen-Ying; Chie, Meng-Chun; Hsu, Wen-Hwei; Chang, Chen-Tien

    2002-08-01

    The alpha-amylase from Bacillus sp. strain TS-23 is a secreted starch hydrolase with a domain organization similar to that of other microbial alpha-amylases and an additional functionally unknown domain (amino acids 517-613) in the C-terminal region. By sequence comparison, we found that this latter domain contained a sequence motif typical for raw-starch binding. To investigate the functional role of the C-terminal region of the alpha-amylase of Bacillus sp. strain TS-23, four His(6)-tagged mutants with extensive deletions in this region were constructed and expressed in Escherichia coli. SDS-PAGE and activity staining analyses showed that the N- and C-terminally truncated alpha-amylases had molecular masses of approximately 65, 58, 54, and 49 kDa. Progressive loss of raw-starch-binding activity occurred upon removal of C-terminal amino acid residues, indicating the requirement for the entire region in formation of a functional starch-binding domain. Up to 98 amino acids from the C-terminal end of the alpha-amylase could be deleted without significant effect on the raw-starch hydrolytic activity or thermal stability. Furthermore, the active mutants hydrolyzed raw corn starch to produce maltopentaose as the main product, suggesting that the raw-starch hydrolytic activity of the Bacillus sp. strain TS-23 alpha-amylase is functional and independent from the starch-binding domain.

  11. Toxicity of radiation-resistant strains of Bacillus thuringiensis (Berl. ) to larval Plutella xylostella (L. )

    Energy Technology Data Exchange (ETDEWEB)

    Jangi, M.S.; Ibrahim, H. (Faculty of Health Sciences, Universiti Kebangsaan, Malysia, Bangi, Selangor)

    1983-05-01

    A total of 24 isolates of Bacillus thuringiensis (Berliner), resistant to a ..gamma..-radiation dose of 100 krad, were screened for their toxicity to larval silkworms, Bombyxmori(L.), and 15 of them were subsequently tested for their toxicity to larval diamond-back moth, Plutella xylostella(L.). The LC/sub 50/'s of these isolates to B. mori ranged from 1.6 X 10/sup 5/ to 6.0 X 10/sup 3/ spores/mL or from 5.9 to 0.3 ..mu..g cellular protein/mL. The irradiation treatment produced isolates which were significantly more toxic to P. xylostella (LC/sub 50/ < 8.1 X 10/sup 4/ spores/mL or 3.7 ..mu..g cellular protein/mL) and/ or less toxic to B. mori (LC/sub 50/ > 2.3 X 10/sup 4/ spores/mL or 1.0 ..mu..g cellular protein/mL) than the parent commercial strain.

  12. Analysis of the sporicidal activity of chlorine dioxide disinfectant against Bacillus anthracis (Sterne strain)

    Science.gov (United States)

    Chatuev, B.A.; Peterson, J.W.

    2009-01-01

    Summary Routine surface decontamination is an essential hospital and laboratory procedure, but the list of effective, noncorrosive disinfectants that kill spores is limited. We investigated the sporicidal potential of an aqueous chlorine dioxide solution and encountered some unanticipated problems. Quantitative bacteriological culture methods were used to determine the log10 reduction of Bacillus anthracis (Sterne strain) spores following 3 min exposure to various concentrations of aqueous chlorine dioxide solutions at room temperature in sealed tubes, as well as spraying onto plastic and stainless steel surfaces in a biological safety cabinet. Serial 10-fold dilutions of the treated spores were then plated on 5% sheep blood agar plates, and the survivor colonies were enumerated. Disinfection of spore suspensions with aqueous chlorine dioxide solution in sealed microfuge tubes was highly effective, reducing the viable spore counts by 8 log10 in only 3 min. By contrast, the process of spraying or spreading the disinfectant onto surfaces resulted in only a 1 log10 kill because the chlorine dioxide gas was rapidly vaporised from the solutions. Full potency of the sprayed aqueous chlorine dioxide solution was restored by preparing the chlorine dioxide solution in 5% bleach (0.3% sodium hypochlorite). The volatility of chlorine dioxide can cause treatment failures that constitute a serious hazard for unsuspecting users. Supplementation of the chlorine dioxide solution with 5% bleach (0.3% sodium hypochlorite) restored full potency and increased stability for one week. PMID:20061062

  13. Biochemical analysis and investigation on the prospective applications of alkaline protease from a Bacillus cereus strain.

    Science.gov (United States)

    Saleem, Mahjabeen; Rehman, Atiqa; Yasmin, Riffat; Munir, Bushra

    2012-06-01

    Proteases have prospective financial and environment-friendly applications; hence attention is focused currently on the finding of new protease producing microorganism so as to meet the requirements of industry. A thermophilic bacterial strain producing extracellular protease activity was isolated from soil and identified as Bacillus cereus by analysis of 16S rRNA. Protease production by the microorganism was improved by studying the impact of the type of nitrogen and carbon source, fermentation period, growth temperature and initial pH of the culture medium in cultivation optimization experiments. The enzyme was purified to homogeneity in two step procedure involving Sephadex G-75 and Q-Sepharose chromatography. The molecular weight of purified enzyme was found to be 58 kDa by SDS-PAGE. Protease exhibited a pH and temperature optima of 7.5 and 60°, respectively. The enzyme was active in the pH range of 6.0-9.0 and stable up to 70°C. Histological analysis of protease treated goat and cow skin pelts showed complete removal of non leather forming structures such as hair shaft, hair follicles and glandular structures. The protease showed the stain removing property from blood stained cotton cloth and found to be compatible with six commercially available detergents. The protease could release peptides from natural proteins after digestion of coagulated egg albumin and blood clot.

  14. Purification and characterization of a novel antifungal protein from Bacillus subtilis strain B29"

    Institute of Scientific and Technical Information of China (English)

    Jing LI; Qian YANG; Li-hua ZHAO; Shu-mei ZHANG; Yu-xia WANG; Xiao-yu ZHAO

    2009-01-01

    An antifungal protein was isolated from a culture of Bacillus subtilis strain B29. The isolation procedure comprised ion exchange chromatography on diethylaminoethyl (DEAE)-52 cellulose and gel filtration chromatography on Bio-Gel P-100.The protein was absorbed on DEAE-cellulose and Bio-Gel P-100. The purified antifungal fraction was designated as B29I, with a molecular mass of 42.3 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), pl value 5.69 by isoelectric focusing (IEF)-PAGE, and 97.81% purity by high performance liquid chromatography (HPLC). B29I exhibited in-hibitory activity on mycelial growth in Fusarium oxysporum, Rhizoctonia solani, Fusarium moniliforme, and Sclerotinia scle-rotiorum. The 50% inhibitory concentrations (IC50) of its antifungal activity toward Fusarium oxysporum and Rhizoctonia solani were 45 and 112 μmol/L, respectively. B291 also demonstrated an inhibitory effect on conidial spore germination of Fusarium oxysporum and suppression of germ-tube elongation, and induced distortion, tumescence, and rupture of a portion of the germi-nated spores.

  15. Characterization of Insecticidal Genes of Bacillus thuringiensis Strains Isolated from Arid Environments.

    Science.gov (United States)

    Abulreesh, Hussein H; Osman, Gamal E H; Assaeedi, Abdulrahman S A

    2012-09-01

    This study aimed at characterizing the insecticidal genes of eight Bacillus thuringiensis isolates that were recovered from the local environment of western Saudi Arabia. The screening for the presence of lepidopteran-specific cry1A family and vip3A genes, dipteran-specific cry4 family and coleopteran-specific cry3A, vip1A and vip2A genes, was carried out by PCR. All eight isolates produced PCR products that confirmed the presence of cry1Aa, cry1Ab, cry1Ac, cry4A, cry4B genes, but not cry3A, vip1A and vip2A genes. However, three isolates only were found to carry vip3A genes as revealed by PCR. The observation of cry1 and cry4 genes suggests that these eight isolates may have dual activity against Lepidoptera and Diptera species, while three isolates possessed vip3 genes in addition to cry1 and cry4 which suggests that these three isolates have toxic crystals and vegetative proteins. The results of this study are interesting in the sense that they may help developing new strategies for controlling insects of economic and medical importance in Saudi Arabia, using B. thuringiensis strains that naturally exist in the local environment instead of the current control strategies that are based solely on chemical insecticides.

  16. Production of keratinolytic enzyme by an indigenous feather-degrading strain Bacillus cereus Wu2.

    Science.gov (United States)

    Lo, Wei-Hsun; Too, Jui-Rze; Wu, Jane-Yii

    2012-12-01

    A novel feather-degrading microorganism was isolated from a poultry farm in Taiwan, and was identified Bacillus cereus Wu2 according to 16S rRNA sequencing. The isolated strain produces keratinolytic enzyme using chicken feather as the sole carbon and nitrogen source. The experimental results indicated that the extra carbon sources (glucose, fructose, starch, sucrose, or lactose) could act as a catabolite repressor to the enzyme secretion or keratinolytic activity when keratinous substrates were employed as protein sources. However, addition of 2 g/L of NH(4)Cl to the feather medium increased the enzyme production. The optimum temperature and initial pH for enzyme production were 30°C and 7.0, respectively. The maximum yield of the enzyme was 1.75 kU/mL in the optimal chicken feather medium; this value was about 17-fold higher than the yield in the basal hair medium. The B. cereus Wu2 possessed disulfide reductase activity along with keratinolytic activity. The amino acid contents of feathers degradated by B. cereus Wu2 were higher, especially for lysine, methionine and threonine which were nutritionally essential amino acids and usually deficient in the feather meal. Thus, B. cereus Wu2 could be not only used to enhance the nutritional value of feather meal but is also a potential bioinoculant in agricultural environments.

  17. Identification of the immunogenic spore and vegetative proteins of Bacillus anthracis vaccine strain A16R.

    Directory of Open Access Journals (Sweden)

    Xiankai Liu

    Full Text Available Immunoproteomics was used to screen the immunogenic spore and vegetative proteins of Bacillus anthracis vaccine strain A16R. The spore and vegetative proteins were separated by 2D gel electrophoresis and transferred to polyvinylidene difluoride membranes, and then western blotting was performed with rabbit immune serum against B.anthracis live spores. Immunogenic spots were cut and digested by trypsin. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry was performed to identify the proteins. As a result, 11 and 45 immunogenic proteins were identified in the spores and vegetative cells, respectively; 26 of which have not been reported previously. To verify their immunogenicity, 12 of the identified proteins were selected to be expressed, and the immune sera from the mice vaccinated by the 12 expressed proteins, except BA0887, had a specific western blot band with the A16R whole cellular lytic proteins. Some of these immunogenic proteins might be used as novel vaccine candidates themselves or for enhancing the protective efficacy of a protective-antigen-based vaccine.

  18. Ability of Bacillus mucilaginosus GY03 Strain to Adsorb Chromium Ions

    Institute of Scientific and Technical Information of China (English)

    CHEN Ye; LIAN Bin

    2005-01-01

    A research with Bacillus mucilaginosus cultured in nitrogen-free medium for forming a flocculant material to adsorb Cr+6 was conducted to determine the effects of different pH, volume, treatment time, and chromium (Ⅵ) concentrations on chromium (Ⅵ) adsorption by microbial flocculant (MBF), which was produced from the B. mucilaginosus GY03 strain. The results showed that MBF had outstanding flocculation on chromium (Ⅵ). Based on the results of a oneway experiment and actual wastewater treatment conditions, the optimum conditions, obtained by using orthogonal experiments, for chromium (Ⅵ) adsorption by MBF were: Cr6+ solution pH of 9, flocculant material volume of 15 mL,treatment time of 12 h and chromium ion concentration of 30 mg L-1. The results demonstrated that the MBF produced from GY03 could be used in the chromium-containing wastewater treatment. Meanwhile, after extraction and analysis of the MBF polysaccharides, it was found that MBF was mainly composed of glycoprotein. Analysis on constituents of monosaccharide showed that polysaccharides of B. mucilaginosus were composed of rhamnose, glucose etc. Thus, because it was applied over a wide range of pH, in small amounts and had a rapid flocculation speed the flocculant used in this experiment had a vast field of application potential.

  19. Purification and characterization of keratinase from a new Bacillus subtilis strain

    Institute of Scientific and Technical Information of China (English)

    Cheng-gang CAI; Ji-shuang CHEN; Jiong-jiong QI; Yun YIN; Xiao-dong ZHENG

    2008-01-01

    The aim of this study was to purify and characterize a keratinase produced by a new isolated Bacillus subtilis KD-N2strain. The keratinase produced by the isolate was purified using ammonium sulphate precipitation, Sephadex G-75 and DEAE (diethylaminoethyl)-Sepharose chromatographic techniques. The purified enzyme was shown to have a molecular mass of 30.5kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The optimum pH at 50℃ was 8.5 and the optimum temperature at pH 8.5 was 55℃. The keratinase was partially inactivated by some metal ions, organic solvents and serine protease inhibitor phenylmethanesulfonyl fluoride (PMSF). Sodium dodecyl sulfate (SDS) and ethylene diamine tetraacetic acid (EDTA) had positive effect on the keratinase activity. Reducing agents including dithiothreitol (DTT),mercaptoethanol, L-cysteine, sodium sulphite, as well as chemicals of SDS, ammonium sulfamate and dimethylsulfoxide (DMSO)stimulated the enzyme activity upon a feather meal substrate. Besides feather keratin, the enzyme is active upon the soluble proteins ovalbumin, bovine serum albumin (BSA), casein and insoluble ones as sheep wool and human hair. Calf hair, silk and collagen could not be hydrolyzed by the keratinase.

  20. Proteome response of Tribolium castaneum larvae to Bacillus thuringiensis toxin producing strains.

    Directory of Open Access Journals (Sweden)

    Estefanía Contreras

    Full Text Available Susceptibility of Tribolium castaneum (Tc larvae was determined against spore-crystal mixtures of five coleopteran specific and one lepidopteran specific Bacillus thuringiensis Cry toxin producing strains and those containing the structurally unrelated Cry3Ba and Cry23Aa/Cry37Aa proteins were found toxic (LC(50 values 13.53 and 6.30 µg spore-crystal mixture/µL flour disc, respectively. Using iTRAQ combined with LC-MS/MS allowed the discovery of seven novel differentially expressed proteins in early response of Tc larvae to the two active spore-crystal mixtures. Proteins showing a statistically significant change in treated larvae compared to non-intoxicated larvae fell into two major categories; up-regulated proteins were involved in host defense (odorant binding protein C12, apolipophorin-III and chemosensory protein 18 and down-regulated proteins were linked to metabolic pathways affecting larval metabolism and development (pyruvate dehydrogenase Eα subunit, cuticular protein, ribosomal protein L13a and apolipoprotein LI-II. Among increased proteins, Odorant binding protein C12 showed the highest change, 4-fold increase in both toxin treatments. The protein displayed amino acid sequence and structural homology to Tenebrio molitor 12 kDa hemolymph protein b precursor, a non-olfactory odorant binding protein. Analysis of mRNA expression and mortality assays in Odorant binding protein C12 silenced larvae were consistent with a general immune defense function of non-olfactory odorant binding proteins. Regarding down-regulated proteins, at the transcriptional level, pyruvate dehydrogenase and cuticular genes were decreased in Tc larvae exposed to the Cry3Ba producing strain compared to the Cry23Aa/Cry37Aa producing strain, which may contribute to the developmental arrest that we observed with larvae fed the Cry3Ba producing strain. Results demonstrated a distinct host transcriptional regulation depending upon the Cry toxin treatment. Knowledge

  1. Identification and characterization of a novel class of extracellular poly(3-hydroxybutyrate) depolymerase from Bacillus sp. strain NRRL B-14911.

    Science.gov (United States)

    Ma, Wan-Ting; Lin, Ju-Hui; Chen, Hui-Ju; Chen, Syuan-Yi; Shaw, Gwo-Chyuan

    2011-11-01

    The catalytic, linker, and denatured poly(3-hydroxybutyrate) (dPHB)-binding domains of bacterial extracellular PHB depolymerases (PhaZs) are classified into several different types. We now report a novel class of extracellular PHB depolymerase from Bacillus sp. strain NRRL B-14911. Its catalytic domain belongs to type 1, whereas its putative linker region neither possesses the sequence features of the three known types of linker domains nor exhibits significant amino acid sequence similarity to them. Instead, this putative linker region can be divided into two distinct linker domains of novel types: LD1 and LD2. LD1 shows significant amino acid sequence similarity to certain regions of a large group of PHB depolymerase-unrelated proteins. LD2 and its homologs are present in a small group of PhaZs. The remaining C-terminal portion of this PhaZ can be further divided into two distinct domains: SBD1 and SBD2. Each domain showed strong binding to dPHB, and there is no significant sequence similarity between them. Each domain neither possesses the sequence features of the two known types of dPHB-binding domains nor shows significant amino acid sequence similarity to them. These unique features indicate the presence of two novel and distinct types of dPHB-binding domains. Homologs of these novel domains also are present in the extracellular PhaZ of Bacillus megaterium and the putative extracellular PhaZs of Bacillus pseudofirmus and Bacillus sp. strain SG-1. The Bacillus sp. NRRL B-14911 PhaZ appears to be a representative of a novel class of extracellular PHB depolymerases.

  2. Rapid focused sequencing: a multiplexed assay for simultaneous detection and strain typing of Bacillus anthracis, Francisella tularensis, and Yersinia pestis.

    Directory of Open Access Journals (Sweden)

    Rosemary S Turingan

    Full Text Available BACKGROUND: The intentional release of Bacillus anthracis in the United States in 2001 has heightened concern about the use of pathogenic microorganisms in bioterrorism attacks. Many of the deadliest bacteria, including the Class A Select Agents Bacillus anthracis, Francisella tularensis, and Yersinia pestis, are highly infectious via the pulmonary route when released in aerosolized form. Hence, rapid, sensitive, and reliable methods for detection of these biothreats and characterization of their potential impact on the exposed population are of critical importance to initiate and support rapid military, public health, and clinical responses. METHODOLOGY/PRINCIPAL FINDINGS: We have developed microfluidic multiplexed PCR and sequencing assays based on the simultaneous interrogation of three pathogens per assay and ten loci per pathogen. Microfluidic separation of amplified fluorescently labeled fragments generated characteristic electrophoretic signatures for identification of each agent. The three sets of primers allowed significant strain typing and discrimination from non-pathogenic closely-related species and environmental background strains based on amplicon sizes alone. Furthermore, sequencing of the 10 amplicons per pathogen, termed "Rapid Focused Sequencing," allowed an even greater degree of strain discrimination and, in some cases, can be used to determine virulence. Both amplification and sequencing assays were performed in microfluidic biochips developed for fast thermal cycling and requiring 7 µL per reaction. The 30-plex sequencing assay resulted in genotypic resolution of 84 representative strains belonging to each of the three biothreat species. CONCLUSIONS/SIGNIFICANCE: The microfluidic multiplexed assays allowed identification and strain differentiation of the biothreat agents Bacillus anthracis, Francisella tularensis, and Yersinia pestis and clear discrimination from closely-related species and several environmental

  3. Characterization of native Bacillus thuringiensis strains and selection of an isolate active against Spodoptera frugiperda and Peridroma saucia.

    Science.gov (United States)

    Alvarez, Analía; Virla, Eduardo G; Pera, Licia M; Baigorí, Mario D

    2009-12-01

    Twelve Bacillus thuringiensis (Bt) strains, isolated from larvae and soil samples in Argentina, were molecularly and phenotypically characterized and their insecticidal activities against Spodoptera frugiperda and Peridroma saucia were determined. One isolate--Bt RT--produced more than 93% mortality on first instar larvae of both species, which was higher than that produced by the reference strain Bt 4D1. Bt RT carried a different cry gene profile than Bt 4D1. Scanning electron microscopy showed the presence of bipyramidal and cuboidal crystals. Phenotypic characterization revealed lytic enzymes that could contribute to Bt pathogenicity.

  4. Bacillus amyloliquefaciens ssp. plantarum strains as potential protective starter cultures for the production of Bikalga, an alkaline fermented food

    DEFF Research Database (Denmark)

    Compaor, C.S.; Nielsen, D.S.; Sawadogo-Lingani, H.

    2013-01-01

    Aims: To identify and screen dominant Bacillus spp. strains isolated from Bikalga, fermented seeds of Hibiscus sabdariffa for their antimicrobial activities in brain heart infusion (BHI) medium and in a H. sabdariffa seed-based medium. Further, to characterize the antimicrobial substances produced....... They produced several lipopeptide antibiotics and showed good potential for biological control of Bikalga. Significance and Impact of the Study: Pathogenic bacteria often occur in spontaneous food fermentations. This is the first report to identify indigenous B. amyloliquefaciens ssp. plantarum strains...

  5. Efficient production of 2,3-butanediol from corn stover hydrolysate by using a thermophilic Bacillus licheniformis strain.

    Science.gov (United States)

    Li, Lixiang; Li, Kun; Wang, Kai; Chen, Chao; Gao, Chao; Ma, Cuiqing; Xu, Ping

    2014-10-01

    In this study, a thermophilic Bacillus licheniformis strain X10 was newly isolated for 2,3-butanediol (2,3-BD) production from lignocellulosic hydrolysate. Strain X10 could utilize glucose and xylose simultaneously without carbon catabolite repression. In addition, strain X10 possesses high tolerance to fermentation inhibitors including furfural, vanillin, formic acid, and acetic acid. In a fed-batch fermentation, 74.0g/L of 2,3-BD was obtained from corn stover hydrolysate, with a productivity of 2.1g/Lh and a yield of 94.6%. Thus, this thermophilic B. licheniformis strain is a candidate for the development of efficient industrial production of 2,3-BD from corn stover hydrolysate.

  6. Estirpes de Bacillus thuringiensis efetivas contra insetos das ordens Lepidoptera, Coleoptera e Diptera Bacillus thuringiensis strains effective against insects of Lepidoptera, Coleoptera and Diptera orders

    Directory of Open Access Journals (Sweden)

    Lílian Botelho Praça

    2004-01-01

    Full Text Available O objetivo deste trabalho foi selecionar entre 300 estirpes de Bacillus thuringiensis as efetivas simultaneamente contra larvas de Spodoptera frugiperda J.E. Smith e Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae, Anthonomus grandis Boheman (Coleoptera: Curculionidae, Aedes aegypti Linnaeus e Culex quinquefasciatus Say (Diptera: Culicidae. Foram selecionadas duas estirpes de B. thuringiensis, denominadas S234 e S997, que apresentaram atividade contra as três ordens de insetos. As estirpes foram caracterizadas por métodos morfológicos, bioquímicos e moleculares. As mesmas apresentaram duas proteínas principais de 130 e 65 kDa, produtos de reação em cadeia da polimerase de tamanho esperado para a detecção dos genes cry1Aa, cry1Ab, cry1Ac, cry1B e cry2 e cristais bipiramidais, cubóides e esféricos.The aim of this work was to select among 300 strains of Bacillus thuringiensis those which are simultaneously effective against larvae of Spodoptera frugiperda J.E. Smith and Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae, Anthonomus grandis Boheman (Coleoptera: Curculionidae, Aedes aegypti Linnaeus and Culex quinquefasciatus Say (Diptera: Culicidae. Two strains of B. thuringiensis were selected, S234 and S997, which presented activity against those three insect orders. Both strains were characterized by morphological, biochemical and molecular methods. They have presented two main proteins with 130 and 65 kDa, polimerase chain reaction products with expected sizes for detection of the genes cry1Aa, cry1Ab, cry1Ac, cry1B and cry2 and bipiramidal, cubical and spherical crystals.

  7. Degradation of Benzo [a] Pyrene by a novel strain Bacillus subtilis BMT4i (MTCC 9447

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    Madhuri Kaushish Lily

    2009-12-01

    Full Text Available Benzo [a] Pyrene (BaP is a highly recalcitrant, polycyclic aromatic hydrocarbon (PAH with high genotoxicity and carcinogenicity. It is formed and released into the environment due to incomplete combustion of fossil fuel and various anthropogenic activities including cigarette smoke and automobile exhausts. The aim of present study is to isolate bacteria which can degrade BaP as a sole source of carbon and energy. We have isolated a novel strain BMT4i (MTCC 9447 of Bacillus subtilis from automobile contaminated soil using BaP (50 μg /ml as the sole source of carbon and energy in basal salt mineral (BSM medium. The growth kinetics of BMT4i was studied using CFU method which revealed that BMT4i is able to survive in BaP-BSM medium up to 40 days attaining its peak growth (10(29 fold increase in cell number on 7 days of incubation. The BaP degradation kinetics of BMT4i was studied using High Performance Liquid Chromatography (HPLC analysis of BaP biodegradation products. BMT4i started degrading BaP after 24 hours and continued up to 28 days achieving maximum degradation of approximately 84.66 %. The above findings inferred that BMT4i is a very efficient degrader of BaP. To our best of knowledge, this is the first report showing utilization of BaP as a sole source of carbon and energy by bacteria. In addition, BMT4i can degrade a wide range of PAHs including naphthalene, anthracene, and dibenzothiophene therefore, it could serve as a better candidate for bioremediation of PAHs contaminated sites.

  8. Kinetics of Molybdenum Reduction to Molybdenum Blue by Bacillus sp. Strain A.rzi

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    A. R. Othman

    2013-01-01

    Full Text Available Molybdenum is very toxic to agricultural animals. Mo-reducing bacterium can be used to immobilize soluble molybdenum to insoluble forms, reducing its toxicity in the process. In this work the isolation of a novel molybdate-reducing Gram positive bacterium tentatively identified as Bacillus sp. strain A.rzi from a metal-contaminated soil is reported. The cellular reduction of molybdate to molybdenum blue occurred optimally at 4 mM phosphate, using 1% (w/v glucose, 50 mM molybdate, between 28 and 30°C and at pH 7.3. The spectrum of the Mo-blue product showed a maximum peak at 865 nm and a shoulder at 700 nm. Inhibitors of bacterial electron transport system (ETS such as rotenone, sodium azide, antimycin A, and potassium cyanide could not inhibit the molybdenum-reducing activity. At 0.1 mM, mercury, copper, cadmium, arsenic, lead, chromium, cobalt, and zinc showed strong inhibition on molybdate reduction by crude enzyme. The best model that fitted the experimental data well was Luong followed by Haldane and Monod. The calculated value for Luong’s constants pmax, Ks, Sm, and n was 5.88 μmole Mo-blue hr−1, 70.36 mM, 108.22 mM, and 0.74, respectively. The characteristics of this bacterium make it an ideal tool for bioremediation of molybdenum pollution.

  9. Mechanistic and stereochemical studies of glycine oxidase from Bacillus subtilis strain R5.

    Science.gov (United States)

    Jamil, Farrukh; Gardner, Qurra-Tul-Ann Afza; Bashir, Qamar; Rashid, Naeem; Akhtar, Muhammad

    2010-08-31

    Glycine oxidase gene from a strain of Bacillus subtilis was cloned and expressed in Escherichia coli. The purified enzyme was found, by mass spectrometry, to have a protein M(r) of 40763 (value of 40761.6 predicted from DNA sequence) and a FAD prosthetic group M(r) of 785.1 (theoretical value of 785.5). Glycine oxidase optimally catalyzes the conversion of glycine and oxygen into glyoxylate, hydrogen peroxide, and ammonia. Using samples of [2-RS-(3)H(2),2-(14)C]-, [2-R-(3)H,2-(14)C]-, and [2-S-(3)H,2-(14)C]glycine, we found that in the overall process H(Si) is removed. Incubation of the enzyme with [2-RS-(3)H(2),2-(14)C]glycine under anaerobic conditions, when only the reducing half of the reaction can occur, led to the recovery of 98.5% of the original glycine, which had the same (3)H:(14)C ratio as the starting substrate. The primary isotope effect was studied using [2-(2)H(2)]glycine, and we found that the specificity constants, k(cat)/K(M), for the protio and deuterio substrates were 1.46 x 10(3) and 1.05 x 10(2) M(-1) s(-1), respectively. Two alternative mechanisms for FAD-containing oxidases that involve either the intermediacy of a FADH(2)-imino acid complex or an amino acid covalently linked to FAD, formed via a carbanion, have been considered. The current knowledge of the mechanisms is reviewed, and we argue that a mechanism involving the FADH(2)-imino acid complex can be dissected to satisfactorily explain some of puzzling observations for which the carbanion mechanism was originally conceived. Furthermore, our results, together with observations in the literature, suggest that the interaction of glycine with the enzyme occurs within a tight ternary complex, which is protected from the protons of the medium.

  10. Purification and partial characterization of bacillocin 490, a novel bacteriocin produced by a thermophilic strain of Bacillus licheniformis

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    De Felice Maurilio

    2002-04-01

    Full Text Available Abstract Background Applications of bacteriocins as food preservatives have been so far limited, principally because of their low antimicrobial activity in foods. Nisin is the only bacteriocin of significant use, but applications are restricted principally because of its very low activity at neutral or alkaline pH. Thus the isolation of new bacteriocins active in foods is desirable. Results We isolated a Bacillus licheniformis thermophilic strain producing a bacteriocin with some novel features, named here bacillocin 490. This bacteriocin was inactivated by pronase E and proteinase K and was active against closely related Bacillus spp. both in aerobic and in anaerobic conditions. Bactericidal activity was kept during storage at 4°C and was remarkably stable in a wide pH range. The bacteriocin was partially purified by elution after adhesion to cells of the food-isolated strain Bacillus smithii and had a rather low mass (2 KDa. Antimicrobial activity against B. smithii was observed also when this organism was grown in water buffalo milk. Conclusions Bacillocin 490 is a novel candidate as a food anti-microbial agent since it displays its activity in milk, is stable to heat treatment and during storage, is active in a wide pH range and has bactericidal activity also at high temperature. These features may allow the use of bacillocin 490 during processes performed at high temperature and as a complementary antimicrobial agent of nisin against some Bacillus spp. in non-acidic foods. The small size suggests its use on solid foods.

  11. 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.

  12. An efficient process for lactic acid production from wheat straw by a newly isolated Bacillus coagulans strain IPE22.

    Science.gov (United States)

    Zhang, Yuming; Chen, Xiangrong; Luo, Jianquan; Qi, Benkun; Wan, Yinhua

    2014-04-01

    A thermophilic lactic acid (LA) producer was isolated and identified as Bacillus coagulans strain IPE22. The strain showed remarkable capability to ferment pentose, hexose and cellobiose, and was also resistant to inhibitors from lignocellulosic hydrolysates. Based on the strain's promising features, an efficient process was developed to produce LA from wheat straw. The process consisted of biomass pretreatment by dilute sulfuric acid and subsequent SSCF (simultaneous saccharification and co-fermentation), while the operations of solid-liquid separation and detoxification were avoided. Using this process, 46.12 g LA could be produced from 100g dry wheat straw with a supplement of 10 g/L corn steep liquid powder at the cellulase loading of 20 FPU (filter paper activity units)/g cellulose. The process by B. coagulans IPE22 provides an economical route to produce LA from lignocellulose.

  13. Identification of a Marine Bacillus Strain C5 and Parathion-Methyl Degradation Characteristics of the Extracellular Esterase B1

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    Jianhua Hao

    2014-01-01

    Full Text Available A bacterial strain C5 that can produce new type of marine esterase was isolated and screened from marine sludge. According to 16S rRNA sequence analysis and physiological and biochemical experiments, the strain was identified as Bacillus subtilis. A single isozyme with a molecular weight of 86 kDa was observed by SDS-PAGE and native-PAGE. On this basis, the mechanism of esterase B1 secreted by strain C5 degrading parathion-methyl was explored, and the effects of temperature and pH on the degradation rate were investigated. From the results, p-nitrophenol was one of the degradation products of B1 degrading parathion-methyl, and the best degradation effect could be achieved at the temperature of 40°C and the neutral pH value.

  14. Comparison of three Bacillus amyloliquefaciens strains growth behaviour and evaluation of the spoilage risk during bread shelf-life.

    Science.gov (United States)

    Valerio, F; Di Biase, M; Huchet, V; Desriac, N; Lonigro, S L; Lavermicocca, P; Sohier, D; Postollec, F

    2015-02-01

    This study aims at the characterisation of growth behaviour of three strains of Bacillus amyloliquefaciens, isolated from ropy bread (ATCC8473), wheat grain (ISPA-S109.3) and semolina (ISPA-N9.1) to estimate rope spoilage risk in pan bread during shelf-life using the Sym'Previus tool. Cardinal values and growth/no growth boundaries were determined in broth, while artificial spore inoculations were performed in dough for various pan bread recipes to compare experimental counts with in silico growth simulations. Finally, two storage scenarios were tested to determine the probability to reach a spoilage threshold during bread shelf-life. Similarly to the safety criteria fixed for Listeria monocytogenes contamination in foodstuff complying with EC regulation, a potential rope spoilage threshold was arbitrary fixed at 5 log CFU/g for B. amyloliquefaciens. This study further underlines a higher rope spoilage potential of the ISPA strains as compared to the ATCC strain, thus emphasizing the interest to characterise both wild strains and reference strain to account for biological variability. In conclusion, this study showed that available decision making tools which are largely recognized to predict behaviour of pathogenic strains, shall also be used with spoilage strains to help maintain food quality and extend shelf-life.

  15. Contrasting oxidative stress response mechanisms in novel strains of Bacillus isolated from the Mars-analog, Mojave Desert

    Science.gov (United States)

    Lera, M.; Marcu, O.

    2011-12-01

    Environmental conditions that limit the presence of life include ionizing radiation, extreme temperatures, and lack of water. These environments are common in our solar system and may contribute to the lack of apparent life. However, analogous environments here on Earth are host to a multitude of thriving microbial life. In order for microbes to survive in dry deserts, they must be must be able to adapt to transient diurnal and seasonal changes in the environment (water, temperature). To uncover response strategies to environmental stress that may prevent cellular damage and ensure adaptation and survival, two distinct, novel strains of Bacillus were isolated from the Mojave Desert (a Mars analog due to its arid conditions and high incidence of ultraviolet light) and classified by their partial 16S RNA gene sequences. These species, despite being closely related, exhibited radically different phenotypes and contrasting strategies for mitigating stress. The two strains had different growth rates, metabolic capacities and sporulation onset times when challenged by crowding and heat-shock. In response to hydrogen peroxide challenge, the intracellular levels of catalase activity, a peroxide-scavenging enzyme, differed for each strain, and were surprisingly lower than that of a non-desert control species of Bacillus. DNA repair mechanisms were more active in one strain than the other, and one isolate responded with an increase in expression of longevity gene orthologs involved in stress response. After multiple rounds of culturing, the peroxide degradation capacity, as well as the growth and sporulation rates remained constant for each strain, which suggests these are permanent features of each strain rather than transient responses. Taken together, these data uncover a diverse arsenal of response mechanisms employed by closely related species to combat stress. These adaptations may provide environmental-niche specificity and the diversity of life even in a scarce

  16. Cellulase and xylanase productions by isolated Amazon Bacillus strains using soybean industrial residue based solid-state cultivation

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    Heck Júlio X.

    2002-01-01

    Full Text Available In Brazil, a large amount of a fibrous residue is generated as result of soybean (Glycine max protein production. This material, which is rich in hemicellulose and cellulose, can be used in solid state cultivations for the production of valuable metabolites and enzymes. In this work, we studied the bioconversion of this residue by bacteria strains isolated from water and soil collected in the Amazon region. Five strains among 87 isolated bacteria selected for their ability to produce either celullases or xylanases were cultivated on the aforementioned residue. From strain BL62, identified as Bacillus subtilis, it was obtained a preparation showing the highest specific cellulase activity, 1.08 UI/mg protein within 24 hours of growth. Concerning xylanase, the isolate BL53, also identified as Bacillus subtilis, showed the highest specific activity for this enzyme, 5.19 UI/mg protein within 72 hours of cultivation. It has also been observed the production of proteases that were associated with the loss of cellulase and xylanase activities. These results indicated that the selected microorganisms, and the cultivation process, have great biotechnological potential.

  17. Isolation of a Halophilic Bacterium, Bacillus sp. Strain NY-6 for Organic Contaminants Removal in Saline Wastewater on Ship

    Institute of Scientific and Technical Information of China (English)

    Jie Gao; Zhenjiang Yu; Xiaohui Zhang; Dan Zhao; Fangbo Zhao

    2013-01-01

    The objective of this research was to examine if certain strains of Bacillus bacteria,could survive in dry powder products and if so,could the bacteria degrade organic contaminants in saline wastewater on a ship.As part of the study,we isolated 7 domesticated strains named NY1,NY2,…,and NY7,the strain NY6 showed to have the best performance for organic matter degradation and could survive in dry powder more than 3 months.NY6 was identified as Bacillus aerius,based on the morphological and physic-chemical properties.Its optimal growth conditions were as follows:salinity was 2%; temperature was 37℃; pH was in 6.5-7.0; best ratio of C∶ N∶ P was 100∶5∶1.The capability of its dry powder for Chemical Oxygen Demand (COD) removal was 800mg COD/g in synthesized marine wastewater with 2% salinity.The spores in the dry powder were 1.972× 108 g1.

  18. Characterization of a bacteriocin-like substance produced from a novel isolated strain of Bacillus subtilis SLYY-3

    Science.gov (United States)

    Li, Junfeng; Li, Hongfang; Zhang, Yuanyuan; Duan, Xiaohui; Liu, Jie

    2014-12-01

    In the present research, the strain SLYY-3 was isolated from sediments of Jiaozhou Bay, Qingdao, China. The strain SLYY-3, which produced a bacteriocin-like substance (BLS), was characterized to be a strain of Bacillus subtillis by biochemical profiling and 16S rDNA sequence analysis. It is the first time to report that Bacillus subtilis from Jiaozhou Bay sediments could produce a BLS. The BLS of B. subtillis SLYY-3 exhibited strong inhibitory activity against gram-positive bacteria (including Staphylococcus aureus and B. subtillis) and some fungi (including Penicillium glaucum, Aspergillus niger and Aspergillus flavus). The antimicrobial activity was detected from culture in the exponential growth phase and reached its maximum when culture entered into stationary growth phase. It was thermo-tolerant even when being kept at 100°C for 60 min without losing any activity and stable over a wide pH range from 1.0 to 12.0 while being inactivated by proteolytic enzyme and trypsin, indicating the proteinaceous nature of the BLS. The BLS was purified by precipitation with hydrochloric acid (HCl) and gel filteration (Sephadex G-100). SDS-PAGE analysis of the extracellular peptides of SLYY-3 revealed a bacteriocin-like protein with a molecular mass of 66 kDa. Altogether, these characteristics indicate the potential of the BLS for food industry as a protection against pathogenic and spoilage microorganisms.

  19. Complete Genome Sequence of Bacillus thuringiensis subsp. thuringiensis Strain IS5056, an Isolate Highly Toxic to Trichoplusia ni

    Science.gov (United States)

    Murawska, Emilia; Fiedoruk, Krzysztof; Bideshi, Dennis K.

    2013-01-01

    The genome sequence of the entomopathogen Bacillus thuringiensis subsp. thuringiensis strain IS5056 was determined. The chromosome is composed of 5,491,935 bp. In addition, IS5056 harbors 14 plasmids ranging from 6,880 to 328,151 bp, four of which contain nine insecticidal protein genes, cry1Aa3, cry1Ab21, cry1Ba1, cry1Ia14, cry2Aa9, cry2Ab1, vip1, vip2, and vip3Aa10. PMID:23516221

  20. Antagonistic Activities of Volatiles from Four Strains of Bacillus spp. and Paenibacillus spp. Against Soil-Borne Plant Pathogens

    Institute of Scientific and Technical Information of China (English)

    LIU Wei-wei; MU Wei; ZHU Bing-yu; DU You-chen; LIU Feng

    2008-01-01

    The four effective antagonistic Bacillus strains,isolated from the rhizosphere soil of cucumber in a greenhouse,produced antifungal volatiles.These volatiles strongly inhibited the growth of the most tested pathogenic fungi with wide host plants,induced the mycelial morphological abnormalities,and decreased the sclerotoid production of Sclerotinia sclerotiorum in sealed plates.Spores of Botrytis cinerea exposed to these volatiles for 24-48 h in cavity slides cracked and the sporaceous inclusion became brown and effused to the suspension.An interesting phenomenon observed was that all the bacterial volatiles exhibited intense inhibitory activities against the pigment formation of tested pathogenic fungi,including Ascochyta citrullina,Alternaria solani,Alternaria brassicae,and so on.Interactions mediated by microbial volatiles could be widespread in soils,and volatiles may play an important role in reducing disease levels.A phylogenetic analysis based on 16S rDNA sequence placed the four bacteria in three species Paenibacillus polymyxa (BMP-11),Bacillus subtilis (BL02),and Bacillus pumilus (BSH-4 and ZB 13).Through headspace sampling and GC-MS analysis,a rich profile was found from B.subtilis and overlapping volatile patterns could be found among the different species.Studies are under the way to find the possible action mechanisms and to seek the effective application of bacterial volatiles in greenhouse.

  1. Application of statistical experimental design for optimisation of bioinsecticides production by sporeless Bacillus thuringiensis strain on cheap medium.

    Science.gov (United States)

    Ben Khedher, Saoussen; Jaoua, Samir; Zouari, Nabil

    2013-01-01

    In order to overproduce bioinsecticides production by a sporeless Bacillus thuringiensis strain, an optimal composition of a cheap medium was defined using a response surface methodology. In a first step, a Plackett-Burman design used to evaluate the effects of eight medium components on delta-endotoxin production showed that starch, soya bean and sodium chloride exhibited significant effects on bioinsecticides production. In a second step, these parameters were selected for further optimisation by central composite design. The obtained results revealed that the optimum culture medium for delta-endotoxin production consists of 30 g L(-1) starch, 30 g L(-1) soya bean and 9 g L(-1) sodium chloride. When compared to the basal production medium, an improvement in delta-endotoxin production up to 50% was noted. Moreover, relative toxin yield of sporeless Bacillus thuringiensis S22 was improved markedly by using optimised cheap medium (148.5 mg delta-endotoxins per g starch) when compared to the yield obtained in the basal medium (94.46 mg delta-endotoxins per g starch). Therefore, the use of optimised culture cheap medium appeared to be a good alternative for a low cost production of sporeless Bacillus thuringiensis bioinsecticides at industrial scale which is of great importance in practical point of view.

  2. Application of statistical experimental design for optimisation of bioinsecticides production by sporeless Bacillus thuringiensis strain on cheap medium

    Directory of Open Access Journals (Sweden)

    Saoussen Ben Khedher

    2013-09-01

    Full Text Available In order to overproduce bioinsecticides production by a sporeless Bacillus thuringiensis strain, an optimal composition of a cheap medium was defined using a response surface methodology. In a first step, a Plackett-Burman design used to evaluate the effects of eight medium components on delta-endotoxin production showed that starch, soya bean and sodium chloride exhibited significant effects on bioinsecticides production. In a second step, these parameters were selected for further optimisation by central composite design. The obtained results revealed that the optimum culture medium for delta-endotoxin production consists of 30 g L-1 starch, 30 g L-1 soya bean and 9g L-1 sodium chloride. When compared to the basal production medium, an improvement in delta-endotoxin production up to 50% was noted. Moreover, relative toxin yield of sporeless Bacillus thuringiensis S22 was improved markedly by using optimised cheap medium (148.5 mg delta-endotoxins per g starch when compared to the yield obtained in the basal medium (94.46 mg delta-endotoxins per g starch. Therefore, the use of optimised culture cheap medium appeared to be a good alternative for a low cost production of sporeless Bacillus thuringiensis bioinsecticides at industrial scale which is of great importance in practical point of view.

  3. A new black fly isolate of Bacillus thuringiensis autoagglutinating strain highly toxic to Simulium pertinax (Kollar (Diptera, Simuliidae larvae

    Directory of Open Access Journals (Sweden)

    Clara FG Cavados

    2005-11-01

    Full Text Available Formulations containing the entomopathogenic Bacillus thuringiensis serovar israelensis strain IPS-82 has been widely applied for mosquito control around the world. Strain IPS-82 is highly active against Aedes aegypti but less active against other well-known vectors such as Culex quinquefasciatus and Simulium spp. larvae. Eighteen strains of B. thuringiensis were isolated from Simulium pertinax larvae naturally occurring in rivers of Southeast Brazil with one demonstrating special toxic effects. Simulated field tests against S. pertinax larvae showed that the native Brazilian autoagglutinanting B. thuringiensis (LFB-FIOCRUZ 1035 has an LC50 at least 25 times lower than the standard IPS-82 strain. The same bacterial preparation was also tested against Ae. aegypti larvae in laboratory trials and the LC50 values obtained with LFB-FIOCRUZ 1035 were at least three times lower than the one for the IPS 82 strain. The results indicate that this strain is more toxic than the standard B. thuringiensis serovar israelensis (H14 in the two Dipteran species tested. It is noteworthy that differences between LC50 values were more pronounced in S. pertinax larvae, the source of the original isolation.

  4. A novel glutamate transport system in poly(γ-glutamic acid)-producing strain Bacillus subtilis CGMCC 0833.

    Science.gov (United States)

    Wu, Qun; Xu, Hong; Zhang, Dan; Ouyang, Pingkai

    2011-08-01

    Bacillus subtilis CGMCC 0833 is a poly(γ-glutamic acid) (γ-PGA)-producing strain. It has the capacity to tolerate high concentration of extracellular glutamate and to utilize glutamate actively. Such a high uptake capacity was owing to an active transport system for glutamate. Therefore, a specific transport system for L-glutamate has been observed in this strain. It was a novel transport process in which glutamate was symported with at least two protons, and an inward-directed sodium gradient had no stimulatory effect on it. K(m) and V(m) for glutamate transport were estimated to be 67 μM and 152 nmol⁻¹ min⁻¹ mg⁻¹ of protein, respectively. The transport system showed structural specificity and stereospecificity and was strongly dependent on extracellular pH. Moreover, it could be stimulated by Mg²⁺, NH₄⁺, and Ca²⁺. In addition, the glutamate transporter in this strain was studied at the molecular level. As there was no important mutation of the transporter protein, it appeared that the differences of glutamate transporter properties between this strain and other B. subtilis strains were not due to the differences of the amino acid sequence and the structure of transporter protein. This is the first extensive report on the properties of glutamate transport system in γ-PGA-producing strain.

  5. Antibacterial activity and genotypic-phenotypic characteristics of bacteriocin-producing Bacillus subtilis KKU213: potential as a probiotic strain.

    Science.gov (United States)

    Khochamit, Nalisa; Siripornadulsil, Surasak; Sukon, Peerapol; Siripornadulsil, Wilailak

    2015-01-01

    The antimicrobial activity and probiotic properties of Bacillus subtilis strain KKU213, isolated from local soil, were investigated. The cell-free supernatant (CFS) of a KKU213 culture containing crude bacteriocins exhibited inhibitory effects on Gram-positive bacteria, including Bacillus cereus, Listeria monocytogenes, Micrococcus luteus, and Staphylococcus aureus. The antibacterial activity of the CFS precipitated with 40% ammonium sulfate (AS) remained even after treatment at 60 and 100 °C, at pH 4 and 10 and with proteolytic enzymes, detergents and heavy metals. When analyzed by SDS-PAGE and overlaid with the indicator strains B. cereus and S. aureus, the 40% AS precipitate exhibited inhibitory activity on proteins smaller than 10 kDa. However, proteins larger than 25 kDa and smaller than 10 kDa were still observed on a native protein gel. Purified subtilosin A was prepared by Amberlite XAD-16 bead extraction and HPLC and analyzed by Nano-LC-QTOF-MS. Its molecular mass was found to be 3.4 kDa, and it retained its antibacterial activity. These results are consistent with the detection of the anti-listerial subtilosin A gene of the sbo/alb cluster in the KKU213 strain, which is 100% identical to that of B. subtilis subsp. subtilis 168. In addition to stable and cyclic subtilosin A, a mixture of many extracellular antibacterial peptides was also detected in the KKU213 culture. The KKU213 strain produced extracellular amylase, cellulase, lipase and protease, is highly acid-resistant (pH 2) when cultured in inulin and promotes health and reduces infection of intestinally colonized broiler chickens. Therefore, we propose that bacteriocin-producing B. subtilis KKU213 could be used as a potential probiotic strain or protective culture.

  6. 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.

    Science.gov (United States)

    Brul, Stanley; van Beilen, Johan; Caspers, Martien; O'Brien, Andrea; de Koster, Chris; Oomes, Suus; Smelt, Jan; Kort, Remco; Ter Beek, Alex

    2011-04-01

    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 possible intoxication. Similar issues though more pending toward spore toxigenicity are observed for the anaerobic Clostridia. The paper indicates the nature of stress resistance and highlights contemporary molecular approaches to analyze the mechanistic basis of it in Bacilli. A molecular comparison between a laboratory strain and a food borne isolate, very similar at the genomic level to the laboratory strain but generating extremely heat resistant spores, is discussed. The approaches cover genome-wide genotyping, proteomics and genome-wide expression analyses studies. The analyses aim at gathering sufficient molecular information to be able to put together an initial framework for dynamic modelling of spore germination and outgrowth behaviour. Such emerging models should be developed both at the population and at the single spore level. Tools and challenges in achieving the latter are succinctly discussed.

  7. Crystalliferous Bacillus cereus group bacteria from a Maryland hardwood forest are dominated by psychrotolerant strains.

    Science.gov (United States)

    Blackburn, Michael B; Martin, Phyllis A W; Kuhar, Daniel; Farrar, Robert R; Gundersen-Rindal, Dawn E

    2014-08-01

    Crystal-forming bacteria of the Bacillus cereus group were isolated from soil samples collected at different elevations within a mixed hardwood forest in central Maryland, and their phylogenetic relationships determined by multilocus sequence analysis. The vast majority of isolates obtained were associated with two phylogenetic groups known to be psychrotolerant, with very few isolates representing phylogenetic groups more typically associated with Bacillus thuringiensis. Isolates from the psychrotolerant groups were found to grow on solid media at 7 °C. Isolates of 11 highly related, novel sequence types (STs) from the psychrotolerant group that includes Bacillus weihenstephanensis were generally found at higher elevations, and were not associated with soils near streams. Isolates of two related STs from the second psychrotolerant group were nearly always found at the bottoms of ravines near streams, in areas abundant in earthworm castings.

  8. A genetically modified broad-spectrum strain of Bacillus thuringiensis toxic against Holotrichia parallela, Anomala corpulenta and Holotrichia oblita.

    Science.gov (United States)

    Jia, Yanhua; Zhao, Can; Wang, Qinglei; Shu, Changlong; Feng, Xiaojie; Song, Fuping; Zhang, Jie

    2014-02-01

    Cry8Ea1 from Bacillus thuringiensis strain Bt185 has insecticidal activity against Holotrichia parallela. Cry8Ca2 from strain HBF-1 is effective against Anomala corpulenta. Cry8Ga1 from strain HBF-18 is toxic to H. oblita. Given the need to broaden the spectrum of B. thuringiensis, a broad-spectrum coleopteran active strain of B. thuringiensis, BIOT185, engineered to express multiple cry genes, including cry8Ea1, cry8Fa1 and cry8Ca2, was created by homologous recombination introducing the cry8Ca2 into the B. thuringiensis strain Bt185 by Liu et al. (Appl Microbiol Biotechnol 87:243-249, 2010). To further broaden the spectrum, an engineered B. thuringiensis strain BIOT1858G was constructed by introducing the recombinant plasmid pSTK-8G containing cry8Ga1 into the engineered B. thuringiensis strain BIOT185. PCR and Southern blotting demonstrated that the cry8Ga1 gene was transferred to the novel strain BIOT1858G. SDS-PAGE and RT-PCR confirmed the normal expression and transcription of the cry8Ga1 gene in addition to the cry8Ea1, cry8Fa1 and cry8Ca2 genes. Bioassays of BIOT1858G indicated that the recombinant strain broadened the spectrum against not only H. parallela susceptible to the Cry8E protein and A. corpulenta susceptible to the Cry8C protein but also H. oblita susceptible to the Cry8G protein. The pesticide could also decrease the cost of production and field labor.

  9. 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.

  10. Activity of spores and extracellular proteins from six Cry+ strains and a Cry- strain of Bacillus thuringiensis subsp. kurstaki against the western spruce budworm, Choristoneura occidentalis (Lepidoptera: Tortricidae).

    Science.gov (United States)

    Kalmykova, Galina; Burtseva, Ljudmila; Milne, Ross; van Frankenhuyzen, Kees

    2009-05-01

    We characterized insecticidal activity of previously untested strains of Bacillus thuringiensis kurstaki belonging to two crystal serovars (K-1 and K-73) against the western spruce budworm (Choristoneura occidentalis Freeman 1967). By testing various components, we demonstrated that spores play a critical role in the pathogenesis of each strain. Spore-free crystals caused low mortality and purified spores were generally not toxic. The addition of spores to purified protoxin increased toxicity several hundred-fold, regardless of the parental strain from which the spores or protoxins were derived. The crystal and spore components did not account for full insecticidal activity of whole sporulated cultures owing to the toxicity of soluble proteins that are secreted during cell growth. We observed a marked difference in toxicity of secreted proteins between the K-1 and K-73 type strains, with the K-1 preparations causing much higher mortality, mass reduction, and inhibition of pupation. There was a consistent correlation between relative toxicity of secreted protein preparations and the presence and quantity of the Vip3A protein, suggesting that this protein contributes to the virulence of B. thuringiensis subsp. kurstaki in western spruce budworm larvae. However, other virulence factors have to be invoked to explain the synergizing effect of spores from both K-1 and K-73 strains on Cry protein toxicity.

  11. Oil biodegradation by Bacillus strains isolated from the rock of an oil reservoir located in a deep-water production basin in Brazil

    Energy Technology Data Exchange (ETDEWEB)

    Duarte da Cunha, C.; Rosado, A.S.; Seldin, L.; Weid, I. von der [Universidade Federal do Rio de Janeiro (Brazil). Dept. de Microbiologia Geral; Sebastian, G.V. [CENPES, Petrobras, Ilha do Fundao, Rio de Janeiro (Brazil)

    2006-12-15

    Sixteen spore forming Gram-positive bacteria were isolated from the rock of an oil reservoir located in a deep-water production basin in Brazil. These strains were identified as belonging to the genus Bacillus using classical biochemical techniques and API 50CH kits, and their identity was confirmed by sequencing of part of the 16S rRNA gene. All strains were tested for oil degradation ability in microplates using Arabian Light and Marlin oils and only seven strains showed positive results in both kinds of oils. They were also able to grow in the presence of carbazole, n-hexadecane and polyalphaolefin (PAO), but not in toluene, as the only carbon sources. The production of key enzymes involved with aromatic hydrocarbons biodegradation process by Bacillus strains (catechol 1,2-dioxygenase and catechol 2,3-dioxygenase) was verified spectrophotometrically by detection of cis,cis-muconic acid and 2-hydroxymuconic semialdehyde, and results indicated that the ortho ring cleavage pathway is preferential. Furthermore, polymerase chain reaction (PCR) products were obtained when the DNA of seven Bacillus strains were screened for the presence of catabolic genes encoding alkane monooxygenase, catechol 1,2-dioxygenase, and/or catechol 2,3-dioxygenase. This is the first study on Bacillus strains isolated from an oil reservoir in Brazil. (orig.)

  12. Characterization of an antifungal compound produced by Bacillus sp. strain A(5) F that inhibits Sclerotinia sclerotiorum.

    Science.gov (United States)

    Kumar, Ankit; Saini, Sandeep; Wray, Victor; Nimtz, Manfred; Prakash, Anil; Johri, B N

    2012-12-01

    A potential antagonist, Bacillus sp. strain A(5) F was isolated from soybean rhizosphere following in vitro dual plate screening. The bacterium displayed strong inhibitory activity in vitro against soybean stem rot pathogen, Sclerotinia sclerotiorum. The culture supernatant of strain A(5) F completely suppressed the mycelial growth of the pathogen, indicating that suppression was due to the presence of antifungal compounds in the culture filtrate. The culture filtrate also suppressed other phytopathogenic fungi including Fusarium oxysporum and Macrophomina phaseolina, in vitro suggesting a broad spectrum antagonistic activity against fungal pathogens. Chemical extraction followed by chromatographic analysis resulted in two antifungal fractions. The high resolution-electron spin ionization-mass spectrometry (HR-ESI-MS) and Nuclear Magnetic Resonance (1D and 2D(1) H) spectra of these antifungal fractions revealed the presence of antifungal compounds, one of which showed similarity to bacillomycin D.

  13. Biotechnological potential of Bacillus salmalaya 139SI: a novel strain for remediating water polluted with crude oil waste.

    Directory of Open Access Journals (Sweden)

    Salmah Ismail

    Full Text Available Environmental contamination by petroleum hydrocarbons, mainly crude oil waste from refineries, is becoming prevalent worldwide. This study investigates the bioremediation of water contaminated with crude oil waste. Bacillus salamalaya 139SI, a bacterium isolated from a private farm soil in the Kuala Selangor in Malaysia, was found to be a potential degrader of crude oil waste. When a microbial population of 108 CFU ml-1 was used, the 139SI strain degraded 79% and 88% of the total petroleum hydrocarbons after 42 days of incubation in mineral salt media containing 2% and 1% of crude oil waste, respectively, under optimum conditions. In the uninoculated medium containing 1% crude oil waste, 6% was degraded. Relative to the control, the degradation was significantly greater when a bacteria count of 99 × 108 CFU ml-1 was added to the treatments polluted with 1% oil. Thus, this isolated strain is useful for enhancing the biotreatment of oil in wastewater.

  14. Complete genome sequence of Bacillus thuringiensis serovar alesti BGSC 4C1, a typical strain with toxicity to Lepidoptera insects.

    Science.gov (United States)

    Wang, Yueying; Fu, Jingjing; Zhu, Qian; Zhu, Lei; Zheng, Jinshui; Liu, Hualin; Peng, Donghai; Ruan, Lifang; Sun, Ming

    2016-12-10

    Bacillus thuringiensis serovar alesti was used to control caterpillars from 1970s. Here we reported the complete genome of BGSC 4C1, the type strain of this serovar. It has a circular chromosome and six plasmids. The largest plasmid pBMB267 contains five insecticidal crystal protein genes (two copies of cry1Ae, cry1Gb, cry2Ab, and a novel cry1M-type gene) and three vegetative insecticidal protein genes (a novel binary toxin gene operon vip1-vip2 and vip3Aa). Besides, the strain also has many genes encodeing virulence factors, and some secondary metabolite biosynthetic gene clusters involved in synthesis of antimicrobial peptides and bacteriocins. In addition, there is a poly γ-glutamate synthesis gene cluster, whose product is a candidate to control inflammasome-mediated disorders and potential in many other fields.

  15. Isolation and characterization of Bacillus subtilis EB-28, an endophytic bacterium strain displaying biocontrol activity against Botrytis cinerea Pers

    Institute of Scientific and Technical Information of China (English)

    Shutong WANG; Tongle HU; Yanling JIAO; Jianjian WEI; Keqiang CAO

    2009-01-01

    The fungal pathogen Botrytis cinerea Pers. causes severe rotting on tomato fruits during storage and shelf life. As a biological control agent, endophytic bacterium was regarded as an effective alternative to chemical control. Out of 238 endophytic bacterial isolates, three strains (EB-15, EB-28, and EB-122) isolated from Lycopersicum esculentum Mill., Speranskia tuberculata (Bge.) Baill, and Dictamnus dasycarpus Turcz. respectively were found to be strongly antagonistic to the pathogen in vitro and were selected for further in vivo tests. One endophytic bacterium strain, encoded EB-28, was selected from the three in vivo tested isolates. The inhibitive rate of EB-28 reached 71.1% in vitro and 52.4% in vivo. EB-28 was identified as Bacillus subtilis according to its morphological, physiological, and biochemical characteristics and 16S rDNA sequence analysis.

  16. Influence of Bacillus spp. strains on seedling growth and physiological parameters of sorghum under moisture stress conditions.

    Science.gov (United States)

    Grover, Minakshi; Madhubala, R; Ali, Sk Z; Yadav, S K; Venkateswarlu, B

    2014-09-01

    Microorganisms isolated from stressed ecosystem may prove as ideal candidates for development of bio-inoculants for stressed agricultural production systems. In the present study, moisture stress tolerant rhizobacteria were isolated from the rhizosphere of sorghum, pigeonpea, and cowpea grown under semiarid conditions in India. Four isolates KB122, KB129, KB133, and KB142 from sorghum rhizosphere exhibited plant growth promoting traits and tolerance to salinity, high temperature, and moisture stress. These isolates were identified as Bacillus spp. by 16S rDNA sequence analysis. The strains were evaluated for growth promotion of sorghum seedlings under two different moisture stress conditions (set-I, continuous 50% soil water holding capacity (WHC) throughout the experiment and set-II, 75% soil WHC for 27 days followed by no irrigation for 5 days) under greenhouse conditions. Plate count and scanning electron microscope studies indicated successful root surface colonization by inoculated bacteria. Plants inoculated with Bacillus spp. strains showed better growth in terms of shoot length and root biomass with dark greenish leaves due to high chlorophyll content while un-inoculated plants showed rolling of the leaves, stunted appearance, and wilting under both stress conditions. Inoculation also improved leaf relative water content and soil moisture content. However, variation in proline and sugar content in the different treatments under two stress conditions indicated differential effect of microbial treatments on plant physiological parameters under stress conditions.

  17. Biosorption Potential of Bacillus salmalaya Strain 139SI for Removal of Cr(VI from Aqueous Solution

    Directory of Open Access Journals (Sweden)

    Arezoo Dadrasnia

    2015-12-01

    Full Text Available The present study investigated the biosorption capacity of live and dead cells of a novel Bacillus strain for chromium. The optimum biosorption condition was evaluated in various analytical parameters, including initial concentration of chromium, pH, and contact time. The Langmuir isotherm model showed an enhanced fit to the equilibrium data. Live and dead biomasses followed the monolayer biosorption of the active surface sites. The maximum biosorption capacity was 20.35 mg/g at 25 °C, with pH 3 and contact time of 50 min. Strain 139SI was an excellent host to the hexavalent chromium. The biosorption kinetics of chromium in the dead and live cells of Bacillus salmalaya (B. salmalaya 139SI followed the pseudo second-order mechanism. Scanning electron microscopy and fourier transform infrared indicated significant influence of the dead cells on the biosorption of chromium based on cell morphological changes. Approximately 92% and 70% desorption efficiencies were achieved using dead and live cells, respectively. These findings demonstrated the high sorption capacity of dead biomasses of B. salmalaya 139SI in the biosorption process. Thermodynamic evaluation (ΔG0, ΔH0, and ΔS0 indicated that the mechanism of Cr(VI adsorption is endothermic; that is, chemisorption. Results indicated that chromium accumulation occurred in the cell wall of B. salmalaya 139SI rather than intracellular accumulation.

  18. Screening of biosurfactant-producing Bacillus strains using glycerol from the biodiesel synthesis as main carbon source.

    Science.gov (United States)

    Sousa, M; Melo, V M M; Rodrigues, S; Sant'ana, H B; Gonçalves, L R B

    2012-08-01

    Glycerol, a co-product of biodiesel production, was evaluated as carbon source for biosurfactant production. For this reason, seven non-pathogenic biosurfactant-producing Bacillus strains, isolated from the tank of chlorination at the Wastewater Treatment Plant at Federal University of Ceara, were screened. The production of biosurfactant was verified by determining the surface tension value, as well as the emulsifying capacity of the free-cell broth against soy oil, kerosene and N-hexadecane. Best results were achieved when using LAMI005 and LAMI009 strains, whose biosurfactant reduced the surface tension of the broth to 28.8 ± 0.0 and 27.1 ± 0.1 mN m(-1), respectively. Additionally, at 72 h of cultivation, 441.06 and 267.56 mg L(-1) of surfactin were produced by LAMI005 and LAMI009, respectively. The biosurfactants were capable of forming stable emulsions with various hydrocarbons, such as soy oil and kerosene. Analyses carried out with high performance liquid chromatography (HPLC) showed that the biosurfactant produced by Bacillus subtilis LAMI009 and LAMI005 was compatible with the commercially available surfactin standard. The values of minimum surface tension and the CMC of the produced biosurfactant indicated that it is feasible to produce biosurfactants from a residual and renewable and low-cost carbon source, such as glycerol.

  19. Lipase-Secreting Bacillus Species in an Oil-Contaminated Habitat: Promising Strains to Alleviate Oil Pollution.

    Science.gov (United States)

    Lee, Li Pin; Karbul, Hudzaifah Mohamed; Citartan, Marimuthu; Gopinath, Subash C B; Lakshmipriya, Thangavel; Tang, Thean-Hock

    2015-01-01

    Lipases are of great interest for different industrial applications due to their diversity and versatility. Among different lipases, microbial lipases are preferable due to their broad substrate specificity, and higher stability with lower production costs compared to the lipases from plants and animals. In the past, a vast number of bacterial species have been reported as potential lipases producers. In this study, the lipases-producing bacterial species were isolated from an oil spillage area in the conventional night market. Isolated species were identified as Bacillus species by biochemical tests which indicate their predominant establishment, and further screened on the agar solid surfaces using lipid and gelatin as the substrates. Out of the ten strains tested, four potential strains were subjected to comparison analysis of the lipolytic versus proteolytic activities. Strain 10 exhibited the highest lipolytic and proteolytic activity. In all the strains, the proteolytic activity is higher than the lipolytic activity except for strain 8, suggesting the possibility for substrate-based extracellular gene induction. The simultaneous secretion of both the lipase and protease is a mean of survival. The isolated bacterial species which harbour both lipase and protease enzymes could render potential industrial-based applications and solve environmental issues.

  20. Lipase-Secreting Bacillus Species in an Oil-Contaminated Habitat: Promising Strains to Alleviate Oil Pollution

    Directory of Open Access Journals (Sweden)

    Li Pin Lee

    2015-01-01

    Full Text Available Lipases are of great interest for different industrial applications due to their diversity and versatility. Among different lipases, microbial lipases are preferable due to their broad substrate specificity, and higher stability with lower production costs compared to the lipases from plants and animals. In the past, a vast number of bacterial species have been reported as potential lipases producers. In this study, the lipases-producing bacterial species were isolated from an oil spillage area in the conventional night market. Isolated species were identified as Bacillus species by biochemical tests which indicate their predominant establishment, and further screened on the agar solid surfaces using lipid and gelatin as the substrates. Out of the ten strains tested, four potential strains were subjected to comparison analysis of the lipolytic versus proteolytic activities. Strain 10 exhibited the highest lipolytic and proteolytic activity. In all the strains, the proteolytic activity is higher than the lipolytic activity except for strain 8, suggesting the possibility for substrate-based extracellular gene induction. The simultaneous secretion of both the lipase and protease is a mean of survival. The isolated bacterial species which harbour both lipase and protease enzymes could render potential industrial-based applications and solve environmental issues.

  1. Total Protein of Crude Extracts and Quantification the Native Bacillus Thuringiensis Strains Isolated From Boyacá and Cundinamarca

    Directory of Open Access Journals (Sweden)

    Eneida Torres Cabra

    2013-12-01

    Full Text Available The Cry proteins of Bacillus thuringiensis (Bt produce parasporal crystal during sporulation which are toxic against insect larvae of lepidopteran, coleopteran and dipteran. In the present study, 10 native isolations of Bt strains were quantified from the Native Strains Bank of the Jorge Tadeo Lozano University, isolated from soil in the municipalities of Ráquira, Santa Sofía, Villa de Leyva and Sutamarchán, in Boyacá; and also in Susa, Cundinamarca. Crude extracts of total proteins from the 10 strains incubated in Luria-Bertani medium (LB for 15 days at 37 °C were obtained. For quantification of total protein of the spore crystal mixture was used the Bradford method using a standard pattern a calibration curve with bovine serum albumin (BSA. After 5 minutes, the determination was realized in the spectrophotometer at a wavelength of 595 nm. The brighter blue were presented by the strains ZBUJTL35 and ZBUJTL39, with a concentration of 560.71 and 526.43 mg / ml, respectively. The Bradford method is a simple and fast technique to estimate the protein concentration. The strains will be used to determine the toxicity over Bemisiatabaci larvae.

  2. Divergence in olfactory host plant preference in D. mojavensis in response to cactus host use.

    Science.gov (United States)

    Date, Priya; Dweck, Hany K M; Stensmyr, Marcus C; Shann, Jodi; Hansson, Bill S; Rollmann, Stephanie M

    2013-01-01

    Divergence in host adaptive traits has been well studied from an ecological and evolutionary perspective, but identification of the proximate mechanisms underlying such divergence is less well understood. Behavioral preferences for host plants are often mediated by olfaction and shifts in preference may be accompanied by changes in the olfactory system. In this study, we examine the evolution of host plant preferences in cactophilic Drosophila mojavensis that feeds and breeds on different cacti throughout its range. We show divergence in electrophysiological responses and olfactory behavior among populations with host plant shifts. Specifically, significant divergence was observed in the Mojave Desert population that specializes on barrel cactus. Differences were observed in electrophysiological responses of the olfactory organs and in behavioral responses to barrel cactus volatiles. Together our results suggest that the peripheral nervous system has changed in response to different ecological environments and that these changes likely contribute to divergence among D. mojavensis populations.

  3. Crystalliferous Bacillus cereus group bacteria from a Maryland hardwood forest are dominated by psychrotolerant strains

    Science.gov (United States)

    Crystal forming Bacillus spp. were isolated from soil samples collected at different elevations within a mixed hardwood forest in central Maryland, and their phylogenetic relationships determined by multilocus sequence analysis. The vast majority of isolates obtained were associated with two phylog...

  4. Complete genome sequence of thermophilic Bacillus smithii type strain DSM 4216

    NARCIS (Netherlands)

    Bosma, Elleke F.; Koehorst, Jasper J.; Hijum, van Sacha A.F.T.; Renckens, Bernadet; Vriesendorp, Bastienne; Weijer, van de Tom; Schaap, Peter J.; Vos, de Willem M.; Oost, van der John; Kranenburg, van Richard

    2016-01-01

    Bacillus smithii is a facultatively anaerobic, thermophilic bacterium able to use a variety of sugars that can be derived from lignocellulosic feedstocks. Being genetically accessible, it is a potential new host for biotechnological production of green chemicals from renewable resources. We deter

  5. Purification and characterization of an antimicrobial peptide produced by Bacillus sp. strain P7

    OpenAIRE

    Fernández Soto, Paulina Alexandra

    2014-01-01

    This study reports a potential novel antimicrobial peptide with narrow spectrum activity against S. aureus NCTC 7447, produced by Bacillus tequilensis. Further studies to improve AMP-P7 purification and characterization are requered to establish its potential used in medicine and industry.

  6. Rational design of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 to increase alpha-cyclodextrin production

    NARCIS (Netherlands)

    Uitdehaag, JCM; Penninga, D; van Alebeek, GJWM; Smith, LM; Dijkstra, BW; Dijkhuizen, L

    2000-01-01

    Cyclodextrin glycosyltransferases (CGTase) (EC;2.4.1.19) are extracellular bacterial enzymes that generate cyclodextrins from starch. All known CGTases produce mixtures of alpha, beta, and gamma-cyclodextrins. A maltononaose inhibitor bound to the active site of the CGTase from Bacillus circulans st

  7. Screening for Pseudomonas and Bacillus antagonistic rhizobacteria strains for the biocontrol of Fusarium wilt of chickpea

    Directory of Open Access Journals (Sweden)

    Hannane Abed

    2016-07-01

    Full Text Available The aim of this work is to study the ability of several isolates belonging to Rhizobacteria (Pseudomonas and Bacillus collected from several chickpea growing areas in Algeria, to control the mycelium growth of Fusarium oxysporum f. sp. ciceris. Interesting isolates were characterized for their morphological characteristics, physiological and biochemical activities as potential bio-control agent. Fungal inhibition tests were performed using plate assay and each isolate were tested for the production of protease, cyanide hydrogen, indole acetic acid, antifungal volatile and extracellular compound. According to API 50 CH, we are able to identify six Bacillus species (B. subtilis, B. circulans, B. lentus, B. aneurinilyticus, B. firmus, B. licheniformis; and with API 20NE test we have identified three Pseudomonas species (P. aeruginosa, P. luteola, P. fluorescens. The ability of bacterial isolates was varied in production of Protease, Gelatinase, Amylase, Cellulase, Acid Indole acetic, Lipase, Catalase and Cyanid Hydrogen. This is traduced in different rate of inhibition growth due to various extracellular compounds, where B61 (Bacillus aneurinilyticus and P39 (Pseudomonas luteola and P70 (Pseudomonas fluorescens were the most efficient with 77 and 55.5% respectively, while B39 (Bacillus firmus and P41 (Pseudomonas luteola were the most efficient by volatile compounds with 70.5 and 77.5% respectively. Our results indicate that these bacteria isolates can be used in the biocontrol of Fusarium oxysporum f. sp. ciceris.

  8. Detection and expression of enterotoxin genes in plant-associated strains of Bacillus cereus

    Science.gov (United States)

    Bacillus cereus is an environmental microbe that commonly inhabits plants and soil. Twenty five plant-associated B. cereus isolates were obtained from apple, cacao, tomato, and potato. The isolates were screened for the presence and expression of enterotoxin B (BcET) components of the nonhemolytic e...

  9. Comparative analysis of biofilm formation by Bacillus cereus reference strains and undomesticated food isolates and the effect of free iron.

    Science.gov (United States)

    Hayrapetyan, Hasmik; Muller, Lisette; Tempelaars, Marcel; Abee, Tjakko; Nierop Groot, Masja

    2015-05-04

    Biofilm formation of Bacillus cereus reference strains ATCC 14579 and ATCC 10987 and 21 undomesticated food isolates was studied on polystyrene and stainless steel as contact surfaces. For all strains, the biofilm forming capacity was significantly enhanced when in contact with stainless steel (SS) as a surface as compared to polystyrene (PS). For a selection of strains, the total CFU and spore counts in biofilms were determined and showed a good correlation between CFU counts and total biomass of these biofilms. Sporulation was favoured in the biofilm over the planktonic state. To substantiate whether iron availability could affect B. cereus biofilm formation, the free iron availability was varied in BHI by either the addition of FeCl3 or by depletion of iron with the scavenger 2,2-Bipyridine. Addition of iron resulted in increased air-liquid interface biofilm on polystyrene but not on SS for strain ATCC 10987, while the presence of Bipyridine reduced biofilm formation for both materials. Biofilm formation was restored when excess FeCl3 was added in combination with the scavenger. Further validation of the iron effect for all 23 strains in microtiter plate showed that fourteen strains (including ATCC10987) formed a biofilm on PS. For eight of these strains biofilm formation was enhanced in the presence of added iron and for eleven strains it was reduced when free iron was scavenged. Our results show that stainless steel as a contact material provides more favourable conditions for B. cereus biofilm formation and maturation compared to polystyrene. This effect could possibly be linked to iron availability as we show that free iron availability affects B. cereus biofilm formation.

  10. [Distribution of strains of spore-forming bacteria of the genus Bacillus in the bottom sediments of Lake Khubsugul in Northern Mongolia as an indication of paleoclimate].

    Science.gov (United States)

    Suslova, M Iu; Parfenova, V V; Ziborova, G A; Fedotov, A P

    2009-01-01

    Data on the distribution and abundance of bacteria of the genus Bacillus in the bottom sediments of Lake Khubsugul have shown the predominance of strains that preferred low temperatures. This indicates fairly cold temperature conditions on the territory of the Khubsugul drainage area. On the whole, the dynamics of interchange of minimums and maximums of abundance of bacteria of the genus Bacillus is similar to the global climate fluctuations. Study of the enzymatic activity of pure cultures revealed that most strains studied possessed proteolytic activity; consequently, the dynamics of bacteria development is correlated with the supply of organic nitrogen-containing compositions.

  11. Delineation of Cis-Acting Sequences Required for Expression of Drosophila Mojavensis Adh-1

    Science.gov (United States)

    Bayer, C. A.; Curtiss, S. W.; Weaver, J. A.; Sullivan, D. T.

    1992-01-01

    The control of expression of the Adh-1 gene of Drosophila mojavensis has been analyzed by transforming ADH null Drosophila melanogaster hosts with P element constructs which contain D. mojavensis Adh-1 having deletions of different extent in the 5' and 3' ends. Adh-1 expression in the D. melanogaster hosts is qualitatively similar to expression in D. mojavensis, although expression is quantitatively lower in transformants. Deletions of the 5' end indicate that information required for normal temporal and tissue expression in larvae is contained within 70 bp of the transcription start site. However, deletion constructs to -70 are deficient in ovarian nurse cell expression, whereas the additional upstream sequences present in constructs containing deletions to -257 do support expression in the ovary. Comparison of the nucleotide sequence in the -257 to -70 region of Adh-1 of four species: D. mojavensis and Drosophila arizona, which express Adh-1 in the ovary, and Drosophila mulleri and Drosophila navojoa, which do not, has led to the identification of regions of sequence similarity that correlate with ovary expression. One of these bears a striking similarity to a conserved sequence located upstream of the three heat shock genes that have constitutive ovarian expression and may be an ovarian control element. We have identified an aberrant aspect of Adh-1 expression. In transformants which carry an Adh-1 gene without a functional upstream Adh-2 gene Adh-1 expression continues into the adult stage instead of ceasing at the onset of metamorphosis. In transformants with a functional Adh-2 gene, Adh-1 expression ceases in the third larval instar stage and aberrant expression in the adult stage does not occur. PMID:1317314

  12. Characteristics and phylogeny of Bacillus cereus strains isolated from Maari, a traditional West African food condiment.

    Science.gov (United States)

    Thorsen, Line; Kando, Christine Kere; Sawadogo, Hagrétou; Larsen, Nadja; Diawara, Bréhima; Ouédraogo, Georges Anicet; Hendriksen, Niels Bohse; Jespersen, Lene

    2015-03-02

    Maari is a spontaneously fermented food condiment made from baobab tree seeds in West African countries. This type of product is considered to be safe, being consumed by millions of people on a daily basis. However, due to the spontaneous nature of the fermentation the human pathogen Bacillus cereus occasionally occurs in Maari. This study characterizes succession patterns and pathogenic potential of B. cereus isolated from the raw materials (ash, water from a drilled well (DW) and potash), seed mash throughout fermentation (0-96h), after steam cooking and sun drying (final product) from two production sites of Maari. Aerobic mesophilic bacterial (AMB) counts in raw materials were of 10(5)cfu/ml in DW, and ranged between 6.5×10(3) and 1.2×10(4)cfu/g in potash, 10(9)-10(10)cfu/g in seed mash during fermentation and 10(7) - 10(9) after sun drying. Fifty three out of total 290 AMB isolates were identified as B. cereus sensu lato by use of ITS-PCR and grouped into 3 groups using PCR fingerprinting based on Escherichia coli phage-M13 primer (M13-PCR). As determined by panC gene sequencing, the isolates of B. cereus belonged to PanC types III and IV with potential for high cytotoxicity. Phylogenetic analysis of concatenated sequences of glpF, gmk, ilvD, pta, pur, pycA and tpi revealed that the M13-PCR group 1 isolates were related to B. cereus biovar anthracis CI, while the M13-PCR group 2 isolates were identical to cereulide (emetic toxin) producing B. cereus strains. The M13-PCR group 1 isolates harboured poly-γ-D-glutamic acid capsule biosynthesis genes capA, capB and capC showing 99-100% identity with the environmental B. cereus isolate 03BB108. Presence of cesB of the cereulide synthetase gene cluster was confirmed by PCR in M13-PCR group 2 isolates. The B. cereus harbouring the cap genes were found in potash, DW, cooking water and at 8h fermentation. The "emetic" type B. cereus were present in DW, the seed mash at 48-72h of fermentation and in the final product

  13. cDNAs of aminopeptidase-like protein genes from Plodia interpunctella strains with different susceptibilities to Bacillus thuringiensis toxins.

    Science.gov (United States)

    Zhu, Y C; Kramer, K J; Oppert, B; Dowdy, A K

    2000-03-01

    Aminopeptidase N has been reported to be a Bacillus thuringiensis (Bt) Cry1A toxin-binding protein in several lepidopteran insects. cDNAs of aminopeptidase-like proteins from both Bt-susceptible RC688s and Bt-resistant HD198r strains of the Indianmeal moth, Plodia interpunctella, were cloned and sequenced. They contain 3345 and 3358 nucleotides, respectively, and each has a 3048 bp open reading frame that encodes 1016 amino acids. Putative protein sequences include 10 potential glycosylation sites and a zinc metal binding site motif of HEXXH, which is typical of the active site of zinc-dependent metallopeptidases. Sequence analysis indicated that the deduced protein sequences are most similar to an aminopeptidase from Heliothis virescens with 62% sequence identity and highly similar to three other lepidopteran aminopeptidases from Plutella xylostella, Manduca sexta, Bombyx mori with sequence identities of 51-52%. Four nucleotide differences were observed in the open reading frames that translated into two amino acid differences in the putative protein sequences. Polymerase chain reaction (PCR) confirmed an aminopeptidase gene coding difference between RC688s and HD198r strains of P. interpunctella in the PCR amplification of a specific allele (PASA) using preferential primers designed from a single base substitution. The gene mutation for Asp185-->Glu185 was also confirmed in two additional Bt-resistant P. interpunctella strains. This mutation is located within a region homologous to the conserved Cry1Aa toxin binding regions from Bombyx mori and Plutella xylostella. The aminopeptidase-like mRNA expression levels in the Bt-resistant strain were slightly higher than those in the Bt-susceptible strain. The sequences reported in this paper have been deposited in the GenBank database (accession numbers AF034483 for susceptible strain RC688s and AF034484 for resistant strain HD198r).

  14. Effect of aflatoxin B1 on growth and enzymatic activity of a native strain of Bacillus sp Efecto de la aflatoxina B1 sobre el crecimiento y actividad proteolítica de una cepa nativa de Bacillus sp

    Directory of Open Access Journals (Sweden)

    Márquez Edna Judith

    2004-07-01

    Full Text Available The effect of different aflatoxin B1 (AFAB1 concentrations on alkaline protease growth and enzymatic activity was evaluated; a native strain of alkalophilic Bacillus sp cultivated in CSL (Corn Steep Liquor was used. It was found that the effect of AFAB1 on the strain inhibited its growth and enzymatic activity to 1 ppm, showing that the strain is highly sensible to AFAB1, meaning that medium obtained f rom Colombian corn contaminated with this mycotoxin cannot be easily used. Concentrations less than 0.1 ppm did not affect growth and enzymatic activity. Key words: Bacillus, aflatoxin, alkaline proteases.Se evaluó el efecto de diferentes concentraciones de aflatoxina B1 (AFAB1 sobre el crecimiento y actividad enzimática de proteasas alcalinas de una cepa nativa de Bacillus sp Alcalofílico cultivada en LAM (Licor Agotado de Maíz. Se encontró que la cepa inhibe su crecimiento y actividad enzimática a 1 ppm, lo que demuestra una alta sensibilidad de la cepa evaluada a la AFAB1 e imposibilita utilizar fácilmente medios obtenidos de maíz nacional contaminado con esta micotoxina. Las concentraciones inferiores a 0.1 ppm no tienen ningún efecto sobre el crecimiento y la actividad enzimática. Palabras clave: Bacillus, aflatoxina, proteasas alcalinas.

  15. Disruption of microbial biofilms by an extracellular protein isolated from epibiotic tropical marine strain of Bacillus licheniformis.

    Directory of Open Access Journals (Sweden)

    Devendra H Dusane

    Full Text Available BACKGROUND: Marine epibiotic bacteria produce bioactive compounds effective against microbial biofilms. The study examines antibiofilm ability of a protein obtained from a tropical marine strain of Bacillus licheniformis D1. METHODOLOGY/PRINCIPAL FINDINGS: B. licheniformis strain D1 isolated from the surface of green mussel, Perna viridis showed antimicrobial activity against pathogenic Candida albicans BH, Pseudomonas aeruginosa PAO1 and biofouling Bacillus pumilus TiO1 cultures. The antimicrobial activity was lost after treatment with trypsin and proteinase K. The protein was purified by ultrafiltration and size-exclusion chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF analysis revealed the antimicrobial agent to be a 14 kDa protein designated as BL-DZ1. The protein was stable at 75°C for 30 min and over a pH range of 3.0 to 11.0. The sequence alignment of the MALDI-fingerprint showed homology with the NCBI entry for a hypothetical protein (BL00275 derived from B. licheniformis ATCC 14580 with the accession number gi52082584. The protein showed minimum inhibitory concentration (MIC value of 1.6 µg/ml against C. albicans. Against both P. aeruginosa and B. pumilus the MIC was 3.12 µg/ml. The protein inhibited microbial growth, decreased biofilm formation and dispersed pre-formed biofilms of the representative cultures in polystyrene microtiter plates and on glass surfaces. CONCLUSION/SIGNIFICANCE: We isolated a protein from a tropical marine strain of B. licheniformis, assigned a function to the hypothetical protein entry in the NCBI database and described its application as a potential antibiofilm agent.

  16. Crystalline protein profiling and cry gene detection in Bacillus thuringiensis strains isolated during epizootics in Cydia pomonella L.

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    Konecka Edyta

    2014-12-01

    Full Text Available The composition of Bacillus thuringiensis crystalline inclusions was characterized in 18 strains: 12 isolates were obtained from the intestinal tract of Cydia pomonella larvae during epizootics, 2 isolates were cultured from Leucoma salicis larvae taken from their natural populations, and 4 reference strains. The number and molecular mass of B. thuringiensis crystalline proteins (Cry and Cyt was estimated by the sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE. The crystals contained 1-8 proteins with molecular masses of 36-155 kDa. The toxin profiles differed both quantatively and qualitatively. The B. thuringiensis MPU B9 isolate had the highest number and diversity of Cry toxins. The analysis of crystal composition by SDS-PAGE was insufficient to detect groups and subgroups of Cry proteins. We identified 20 groups and 3 subgroups of Cry and Cyt crystalline toxins. Only one epizootic strain harboured cry25. In single reference strains, the cry1H, cry10 and cry25 genes were found. We did not find any correlation between the occurrence of cry genes and electrophoretic protein profiles of crystalline toxins.

  17. Characterization of endophytic Bacillus strains from tomato plants (Lycopersicon esculentum) displaying antifungal activity against Botrytis cinerea Pers.

    Science.gov (United States)

    Kefi, Asma; Ben Slimene, Imen; Karkouch, Ines; Rihouey, Christophe; Azaeiz, Sana; Bejaoui, Marwa; Belaid, Rania; Cosette, Pascal; Jouenne, Thierry; Limam, Ferid

    2015-12-01

    Eighty endophytic bacteria were isolated from healthy tissues of roots, stems, leaves and fruits of tomato plants (Lycopersicon esculentum). Four strains, named BL1, BT5, BR8 and BF11 were selected for their antagonism against Botrytis cinerea, a phytopathogenic fungus responsible of gray mold in several important crops, with growth inhibitory activity ranging from 27 to 53%. Morphological, biochemical, and molecular parameters as 16S rDNA sequencing demonstrated that the selected bacterial strains were related to Bacillus species which are known to produce and secrete a lot of lipopeptides with strong inhibitory effect against pathogen mycelial growth. Electrospray mass spectrometry analysis showed that these strains produced heterogeneous mixture of antibiotics belonging to fengycin and surfactin for BL1 and BT5, to iturin and surfactin for BR8, to bacillomycin D, fengycin and surfactin for BF11. Furthermore, these bacteria exhibited biocontrol potential by reducing the disease severity when tested on detached leaflets. Based on their antifungal activity against Botrytis cinerea, these strains could be used for biological control of plant diseases.

  18. Reduced levels of membrane-bound alkaline phosphatase are common to lepidopteran strains resistant to Cry toxins from Bacillus thuringiensis.

    Directory of Open Access Journals (Sweden)

    Juan Luis Jurat-Fuentes

    Full Text Available Development of insect resistance is one of the main concerns with the use of transgenic crops expressing Cry toxins from the bacterium Bacillus thuringiensis. Identification of biomarkers would assist in the development of sensitive DNA-based methods to monitor evolution of resistance to Bt toxins in natural populations. We report on the proteomic and genomic detection of reduced levels of midgut membrane-bound alkaline phosphatase (mALP as a common feature in strains of Cry-resistant Heliothis virescens, Helicoverpa armigera and Spodoptera frugiperda when compared to susceptible larvae. Reduced levels of H. virescens mALP protein (HvmALP were detected by two dimensional differential in-gel electrophoresis (2D-DIGE analysis in Cry-resistant compared to susceptible larvae, further supported by alkaline phosphatase activity assays and Western blotting. Through quantitative real-time polymerase chain reaction (qRT-PCR we demonstrate that the reduction in HvmALP protein levels in resistant larvae are the result of reduced transcript amounts. Similar reductions in ALP activity and mALP transcript levels were also detected for a Cry1Ac-resistant strain of H. armigera and field-derived strains of S. frugiperda resistant to Cry1Fa. Considering the unique resistance and cross-resistance phenotypes of the insect strains used in this work, our data suggest that reduced mALP expression should be targeted for development of effective biomarkers for resistance to Cry toxins in lepidopteran pests.

  19. Complete genome sequence and transcriptomic analysis of a novel marine strain Bacillus weihaiensis reveals the mechanism of brown algae degradation.

    Science.gov (United States)

    Zhu, Yueming; Chen, Peng; Bao, Yunjuan; Men, Yan; Zeng, Yan; Yang, Jiangang; Sun, Jibin; Sun, Yuanxia

    2016-11-30

    A novel marine strain representing efficient degradation ability toward brown algae was isolated, identified, and assigned to Bacillus weihaiensis Alg07. The alga-associated marine bacteria promote the nutrient cycle and perform important functions in the marine ecosystem. The de novo sequencing of the B. weihaiensis Alg07 genome was carried out. Results of gene annotation and carbohydrate-active enzyme analysis showed that the strain harbored enzymes that can completely degrade alginate and laminarin, which are the specific polysaccharides of brown algae. We also found genes for the utilization of mannitol, the major storage monosaccharide in the cell of brown algae. To understand the process of brown algae decomposition by B. weihaiensis Alg07, RNA-seq transcriptome analysis and qRT-PCR were performed. The genes involved in alginate metabolism were all up-regulated in the initial stage of kelp degradation, suggesting that the strain Alg07 first degrades alginate to destruct the cell wall so that the laminarin and mannitol are released and subsequently decomposed. The key genes involved in alginate and laminarin degradation were expressed in Escherichia coli and characterized. Overall, the model of brown algae degradation by the marine strain Alg07 was established, and novel alginate lyases and laminarinase were discovered.

  20. Characterization of Tunisian Bacillus thuringiensis strains with abundance of kurstaki subspecies harbouring insecticidal activities against the lepidopteran insect Ephestia kuehniella.

    Science.gov (United States)

    Saadaoui, Imen; Al-Thani, Roda; Al-Saadi, Fatma; Belguith-Ben Hassan, Najeh; Abdelkefi-Mesrati, Lobna; Schultz, Patrick; Rouis, Souad; Jaoua, Samir

    2010-12-01

    The study of 257 crystal-producing Bacillus thuringiensis isolates from bioinsecticide free soil samples collected from different sites in Tunisia, was performed by PCR amplification, using six primer pairs specific for cry1, cry2, cry3, cry4, and vip3A genes, by the investigation of strain plasmid pattern, crystal morphology and delta-endotoxin content and by the assessment of insecticidal activities against the lepidopteran insect Ephestia kuehniella. Based on plasmid pattern study, 11 representative strains of the different classes were subjected to morphological and molecular analyses. The comparison of the PFGE fingerprints confirmed the heterogeneity of these strains. B. thuringiensis kurstaki strains, harbouring at the same time the genes cry1A, cry2, cry1Ia, and vip3A, were the most abundant (65.4%). 33.34% of the new isolates showed particular delta-endotoxin profiles but no PCR products with the used primer sets. B. thuringiensis israelensis was shown to be also very rare among the Tunisian B. thuringiensis isolates diversity. These findings could have considerable impacts for the set up of new pest control biological agents.

  1. Subtilomycin: A New Lantibiotic from Bacillus subtilis Strain MMA7 Isolated from the Marine Sponge Haliclona simulans

    Directory of Open Access Journals (Sweden)

    Teresa M. Barbosa

    2013-06-01

    Full Text Available Bacteriocins are attracting increased attention as an alternative to classic antibiotics in the fight against infectious disease and multidrug resistant pathogens. Bacillus subtilis strain MMA7 isolated from the marine sponge Haliclona simulans displays a broad spectrum antimicrobial activity, which includes Gram-positive and Gram-negative pathogens, as well as several pathogenic Candida species. This activity is in part associated with a newly identified lantibiotic, herein named as subtilomycin. The proposed biosynthetic cluster is composed of six genes, including protein-coding genes for LanB-like dehydratase and LanC-like cyclase modification enzymes, characteristic of the class I lantibiotics. The subtilomycin biosynthetic cluster in B. subtilis strain MMA7 is found in place of the sporulation killing factor (skf operon, reported in many B. subtilis isolates and involved in a bacterial cannibalistic behaviour intended to delay sporulation. The presence of the subtilomycin biosynthetic cluster appears to be widespread amongst B. subtilis strains isolated from different shallow and deep water marine sponges. Subtilomycin possesses several desirable industrial and pharmaceutical physicochemical properties, including activity over a wide pH range, thermal resistance and water solubility. Additionally, the production of the lantibiotic subtilomycin could be a desirable property should B. subtilis strain MMA7 be employed as a probiotic in aquaculture applications.

  2. Subtilomycin: a new lantibiotic from Bacillus subtilis strain MMA7 isolated from the marine sponge Haliclona simulans.

    Science.gov (United States)

    Phelan, Robert W; Barret, Matthieu; Cotter, Paul D; O'Connor, Paula M; Chen, Rui; Morrissey, John P; Dobson, Alan D W; O'Gara, Fergal; Barbosa, Teresa M

    2013-06-03

    Bacteriocins are attracting increased attention as an alternative to classic antibiotics in the fight against infectious disease and multidrug resistant pathogens. Bacillus subtilis strain MMA7 isolated from the marine sponge Haliclona simulans displays a broad spectrum antimicrobial activity, which includes Gram-positive and Gram-negative pathogens, as well as several pathogenic Candida species. This activity is in part associated with a newly identified lantibiotic, herein named as subtilomycin. The proposed biosynthetic cluster is composed of six genes, including protein-coding genes for LanB-like dehydratase and LanC-like cyclase modification enzymes, characteristic of the class I lantibiotics. The subtilomycin biosynthetic cluster in B. subtilis strain MMA7 is found in place of the sporulation killing factor (skf) operon, reported in many B. subtilis isolates and involved in a bacterial cannibalistic behaviour intended to delay sporulation. The presence of the subtilomycin biosynthetic cluster appears to be widespread amongst B. subtilis strains isolated from different shallow and deep water marine sponges. Subtilomycin possesses several desirable industrial and pharmaceutical physicochemical properties, including activity over a wide pH range, thermal resistance and water solubility. Additionally, the production of the lantibiotic subtilomycin could be a desirable property should B. subtilis strain MMA7 be employed as a probiotic in aquaculture applications.

  3. Genomic Signatures of Strain Selection and Enhancement in Bacillus atrophaeus var. globigii, a Historical Biowarfare Simulant

    Science.gov (United States)

    2011-03-25

    included in the heatmap (i.e. this better represents the locations of the phenotypes which correspond to different modes of action categories). The...sensitivity to beta-lactams, quinolones , and membrane-disrupting activities. These results suggested broad combined effects of several mutations on the...although nitric oxide synthesis plays a critical role in modulating antibiotic resistance in Bacillus spp. [39]. In addition to its role in promoting

  4. Partial isolation and some properties of enterotoxin produced by Bacillus cereus strains

    Directory of Open Access Journals (Sweden)

    T. V. Guaycurus

    1993-03-01

    Full Text Available Extracellular proteins produced by Bacillus cereus AL-42 and AL-15 were fractioned by chromatography on QAE-Sephadex and Sephadex G75. This last chromatographic process resulted in three peaks. The major peak showed vascular permeability activity to rabbits, lethality to mice, and cytotoxicity to Vero and Hela cells. The analysis by SDS-PAGE after ultrafiltration confirm recent findings that the enterotoxin is a compound with molecular mass > 30.000.

  5. Bacillus mojavensis biofilm formation and biosurfactant production using a Laser Ablation Electrospray Ionization System

    Science.gov (United States)

    Biofilms are important extracellular polymeric compounds produced by bacteria that are useful for developmental phases including motility, swarming, signaling processes, and for hydrophobic nutrient utilization, all of which are important attributes for endophytic bacteria with biocontrol potential....

  6. Direct starch fermentation to L-lactic acid by a newly isolated thermophilic strain, Bacillus sp. MC-07.

    Science.gov (United States)

    Poudel, Pramod; Tashiro, Yukihiro; Miyamoto, Hirokuni; Miyamoto, Hisashi; Okugawa, Yuki; Sakai, Kenji

    2015-01-01

    A newly isolated Bacillus sp. MC-07 showed 99.2 % 16S rRNA gene sequence similarity with the Bacillus thermoamylovorans LMG 18084(T). It demonstrated optimum and maximum growth temperatures of 50 and 62 °C, respectively. The ability of MC-07 to produce optically pure L-lactic acid via direct fermentation of starch without enzymatic hydrolysis was investigated at different pH values (6.0-8.0) by intermittent adjustments every 12 h. During batch fermentation in mineral salt medium containing 0.001 % yeast extract at pH 7.0, 20 g/L of soluble starch was utilized to produce 16.6 g/L L-lactic acid at 50 °C within 24 h of fermentation, with 100 % optical purity, 92.1 % lactic acid selectivity, and an L-lactic acid yield of 0.977 g/g. Direct starch fermentation at pHs 6.0, 6.5, 7.5, and 8.0 resulted in considerably lower concentrations of lactic acid than did at pH 7.0. Compared with B. thermoamylovorans LMG 18084(T), the ability of strain MC-07 to produce L-lactic acid was superior.

  7. Diversity of Bacillus cereus group strains is reflected in their broad range of pathogenicity and diverse ecological lifestyles.

    Science.gov (United States)

    Ceuppens, Siele; Boon, Nico; Uyttendaele, Mieke

    2013-06-01

    Bacillus cereus comprises a highly versatile group of bacteria, which are of particular interest because of their capacity to cause disease. Emetic food poisoning is caused by the toxin cereulide produced during the growth of emetic B. cereus in food, while diarrhoeal food poisoning is the result of enterotoxin production by viable vegetative B. cereus cells in the small intestine, probably in the mucus layer and/or attached to the host's intestinal epithelium. The numbers of B. cereus causing disease are highly variable, depending on diverse factors linked to the host (age, diet, physiology and immunology), bacteria (cellular form, toxin genes and expression) and food (nutritional composition and meal characteristics). Bacillus cereus group strains show impressive ecological diversity, ranging from their saprophytic life cycle in soil to symbiotic (commensal and mutualistic) lifestyles near plant roots and in guts of insects and mammals to various pathogenic ones in diverse insect and mammalian hosts. During all these different ecological lifestyles, their toxins play important roles ranging from providing competitive advantages within microbial communities to inhibition of specific pathogenic organisms for their host and accomplishment of infections by damaging their host's tissues.

  8. Bacillus amyloliquefaciens strains isolated from moisture-damaged buildings produced surfactin and a substance toxic to mammalian cells.

    Science.gov (United States)

    Mikkola, Raimo; Andersson, Maria A; Grigoriev, Pavel; Teplova, Vera V; Saris, Nils-Erik L; Rainey, Frederick A; Salkinoja-Salonen, Mirja S

    2004-04-01

    Fungicidic Bacillus amyloliquefaciens strains isolated from the indoor environment of moisture-damaged buildings contained heat-stable, methanol-soluble substances that inhibited motility of boar spermatozoa within 15 min of exposure and killed feline lung cells in high dilution in 1 day. Boar sperm cells lost motility, cellular ATP, and NADH upon contact to the bacterial extract (0.2 microg dry wt/ml). Two bioactive substances were purified from biomass of the fungicidal isolates. One partially characterized substance, 1,197 Da, was moderately hydrophobic and contained leucine, proline, serine, aspartic acid, glutamic acid and tyrosine, in addition to chromophore(s) absorbing at 365 nm. In boar sperm and human neural cells (Paju), the compound depolarized the transmembrane potentials of mitochondria (Delta Psi(m)) and the plasma membrane (Delta Psi(p)) after a 20-min exposure and formed cation-selective channels in lipid membranes, with a selectivity K(+):Na(+):Ca(2+) of 26:15:3.5. The other substance was identified as a plasma-membrane-damaging lipopeptide surfactin. Plate-grown biomass of indoor Bacillus amyloliquefaciens contained ca. 7% of dry weight of the two substances, 1,197 Da and surfactin, in a ratio of 1:6 (w:w). The in vitro observed simultaneous collapse of both cytosolic and mitochondrial ATP in the affected mammalian cell, induced by the 1,197-Da cation channel, suggests potential health risks for occupants of buildings contaminated with such toxins.

  9. Genome Sequence of the Mosquitocidal Bacillus thuringiensis Strain BR58, a Biopesticide Product Effective against the Coffee Berry Borer (Hypothenemus hampei)

    Science.gov (United States)

    Zorzetti, Janaina; Ricietto, Ana P. S.; da Silva, Carlos R. M.; Wolf, Ivan R.; Neves, Pedro M. O. J.; Meneguim, Ana M.; Vilas-Boas, Laurival A.

    2015-01-01

    Bacillus thuringiensis is an important microbial control agent against insect pests. The draft genome sequence of the Brazilian strain BR58 described here contains the insecticidal genes cry4A, cry4B, cry10A, cry11A, cry60A, cry60B, and cyt1A, which show toxicity to both Aedes aegypti and Hypothenemus hampei larvae. PMID:26659669

  10. Draft Genome Sequences of Three Cellulolytic Bacillus licheniformis Strains Isolated from Imperial Geyser, Amphitheater Springs, and Whiterock Springs inside Yellowstone National Park

    Science.gov (United States)

    O' Hair, Joshua A.; Li, Hui; Thapa, Santosh; Scholz, Matthew

    2017-01-01

    ABSTRACT Novel cellulolytic microorganisms are becoming more important for rapidly growing biofuel industries. This paper reports the draft genome sequences of Bacillus licheniformis strains YNP2-TSU, YNP3-TSU, and YNP5-TSU. These cellulolytic isolates were collected from several hydrothermal features inside Yellowstone National Park. PMID:28360181

  11. Complete sequence of the first chimera genome constructed by cloning the whole genome of Synechocystis strain PCC6803 into the Bacillus subtilis 168 genome.

    Science.gov (United States)

    Watanabe, Satoru; Shiwa, Yuh; Itaya, Mitsuhiro; Yoshikawa, Hirofumi

    2012-12-01

    Genome synthesis of existing or designed genomes is made feasible by the first successful cloning of a cyanobacterium, Synechocystis PCC6803, in Gram-positive, endospore-forming Bacillus subtilis. Whole-genome sequence analysis of the isolate and parental B. subtilis strains provides clues for identifying single nucleotide polymorphisms (SNPs) in the 2 complete bacterial genomes in one cell.

  12. Draft Genome Sequence of Plant Growth-Promoting Drought-Tolerant Bacillus sp. Strain CMAA 1363 Isolated from the Brazilian Caatinga Biome

    Science.gov (United States)

    Santos, Suikinai Nobre; Taketani, Rodrigo Gouvêa; Vasconcellos, Rafael Leandro Figueiredo; Melo, Itamar Soares

    2017-01-01

    ABSTRACT The strain of Bacillus sp. CMAA 1363 was isolated from the Brazilian Caatinga biome and showed plant growth-promoting traits and ability to promote maize growth under drought stress. Sequencing revealed genes involved in stress response and plant growth promotion. These genomic features might aid in the protection of plants against the negative effects imposed by drought. PMID:28153893

  13. Conclusion on the peer review of the pesticide risk assessment of the active substance Bacillus amyloliquefaciens subsp. plantarum strain D747

    Directory of Open Access Journals (Sweden)

    European Food Safety Authority

    2014-04-01

    Full Text Available The conclusions of the European Food Safety Authority (EFSA following the peer review of the initial risk assessments carried out by the competent authority of the rapporteur Member State Germany, for the pesticide active substance Bacillus amyloliquefaciens subsp. plantarum strain D747 are reported.  The context of the peer review was that required by Commission Regulation (EU No 188/2011.  The conclusions were reached on the basis of the evaluation of the representative uses of Bacillus amyloliquefaciens subsp. plantarum strain D747 as a fungicide on table and wine grapes. The reliable endpoints concluded as being appropriate for use in regulatory risk assessment, derived from the available studies and literature in the dossier peer reviewed, are presented.  Missing information identified as being required by the regulatory framework is listed.  The assessment of potential transfer of genetic material from Bacillus amyloliquefaciens subsp. plantarum strain D747 to other organisms cannot not be finalised.  The consumer and re-entry worker risk assessment cannot be finalised as data are missing on the levels of secondary metabolites/toxins produced by the organism on plants.  Furthermore, the risk assessment for non-target organisms and the groundwater exposure assessment from secondary metabolites/toxins could not be finalised.  As the soil exposure assessments could not be finalised, also the risk to soil-dwelling organisms from Bacillus amyloliquefaciens subsp. plantarum strain D747 could not be finalised.

  14. Bacillus halodurans Strain C125 Encodes and Synthesizes Enzymes from Both Known Pathways To Form dUMP Directly from Cytosine Deoxyribonucleotides

    DEFF Research Database (Denmark)

    Oehlenschlæger, Christian Berg; Løvgreen, Monika Nøhr; Reinauer, Eva

    2015-01-01

    Analysis of the genome of Bacillus halodurans strain C125 indicated that two pathways leading from a cytosine deoxyribonucleotide to dUMP, used for dTMP synthesis, were encoded by the genome of the bacterium. The genes that were responsible, the comEB gene and the dcdB gene, encoding dCMP deaminase...

  15. Isolation and characterization of a β-propeller gene containing phosphobacterium Bacillus subtilis strain KPS-11 for growth promotion of potato (Solanum tuberosum L.)

    NARCIS (Netherlands)

    Hanif, Muhammad Kashif; Hameed, Sohail; Imran, Asma; Naqqash, Tahir; Shahid, Muhammad; van Elsas, Jan D

    2015-01-01

    Phosphate-solubilizing and phytate-mineralizing bacteria collectively termed as phosphobacteria provide a sustainable approach for managing P-deficiency in agricultural soils by supplying inexpensive phosphate to plants. A phosphobacterium Bacillus subtilis strain KPS-11 (Genbank accession no. KP006

  16. 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

  17. Draft Genome Sequence of Bacillus sp. Strain NSP2.1, a Nonhalophilic Bacterium Isolated from the Salt Marsh of the Great Rann of Kutch, India

    Science.gov (United States)

    Pal, Kamal Krishna; Sherathia, Dharmesh; Dalsania, Trupti; Savsani, Kinjal; Patel, Ilaxi; Sukhadiya, Bhoomika; Mandaliya, Mona; Thomas, Manesh; Ghorai, Sucheta; Vanpariya, Sejal; Rupapara, Rupal; Rawal, Priya; Saxena, Anil Kumar

    2013-01-01

    The 5.52-Mbp draft genome sequence of Bacillus sp. strain NSP2.1, a nonhalophilic bacterium isolated from the salt marsh of the Great Rann of Kutch, India, is reported here. An analysis of the genome of this organism will facilitate the understanding of its survival in the salt marsh. PMID:24158559

  18. Draft Genome Sequence of Bacillus sp. Strain NSP9.1, a Moderately Halophilic Bacterium Isolated from the Salt Marsh of the Great Rann of Kutch, India

    Science.gov (United States)

    Pal, Kamal Krishna; Sherathia, Dharmesh; Dalsania, Trupti; Savsani, Kinjal; Patel, Ilaxi; Thomas, Manesh; Ghorai, Sucheta; Vanpariya, Sejal; Rupapara, Rupal; Rawal, Priya; Sukhadiya, Bhoomika; Mandaliya, Mona; Saxena, Anil Kumar

    2013-01-01

    We report the 4.52-Mbp draft genome sequence of Bacillus sp. strain NSP9.1, a moderately halophilic bacterium isolated from the salt marsh of the Great Rann of Kutch, India. Analysis of the genome of this organism will lead to a better understanding of the genes and metabolic pathways involved in imparting osmotolerance. PMID:24115550

  19. Draft Genome Sequence of Bacillus sp. Strain SB47, an Obligate Extreme Halophile Isolated from a Salt Pan of the Little Rann of Kutch, India

    Science.gov (United States)

    Dey, Rinku; Thomas, Manesh; Sherathia, Dharmesh; Dalsania, Trupti; Patel, Ilaxi; Savsani, Kinjal; Ghorai, Sucheta; Vanpariya, Sejal; Sukhadiya, Bhoomika; Mandaliya, Mona; Rupapara, Rupal; Rawal, Priya; Saxena, Anil Kumar

    2013-01-01

    Here, we report the 4.46-Mbp draft genome sequence of Bacillus sp. strain SB47, an extreme halophile isolated from a salt pan of the Little Rann of Kutch, India. Exploring the genome of this organism will facilitate the understanding and isolation of the gene(s) involved in its extreme osmotolerance. PMID:24115544

  20. Draft Genome Sequence of the Extremely Halophilic Bacillus sp. Strain SB49, Isolated from a Salt Crystallizer Pond of the Little Rann of Kutch, India

    Science.gov (United States)

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

    2013-01-01

    Here we report the draft whole-genome sequence (3.72 Mbp) of Bacillus sp. strain SB49, an extremely halophilic bacterium isolated from a salt crystallizer pond of the Little Rann of Kutch in India. Unraveling the genome of this organism will facilitate understanding and isolation of the genes involved in imparting extreme osmotolerance. PMID:24136852

  1. Comparative genomics of iron-transporting systems in Bacillus cereus strains and impact of iron sources on growth and biofilm formation

    NARCIS (Netherlands)

    Hayrapetyan, Hasmik; Siezen, Roland; Abee, Tjakko; Nierop Groot, Masja

    2016-01-01

    Iron is an important element for bacterial viability, however it is not readily available in most environments. We studied the ability of 20 undomesticated food isolates of Bacillus cereus and two reference strains for capacity to use different (complex) iron sources for growth and biofilm format

  2. Screening and Molecular Identification of New Microbial Strains for Production of Enzymes of Biotechnological Interest

    Directory of Open Access Journals (Sweden)

    Imen Ghazala

    Full Text Available ABSTRACT: This research focused on isolation, identification and characterization of new strains of fungi and bacteria, which were able to produce extracellular xylanase, mannanase, pectinase and α-amylase. Fungi isolates were identified on the basis of analyses of 18S gene sequencing and internal transcribed spacer region. The closest phylogenetic neighbors according to 18S gene sequence and ITS region data for the two isolates M1 and SE were Aspergillus fumigatus and Aspergillus sydowii, respectively. I4 was identified as Bacillus mojavensis on the basis of the 16S rRNA gene sequencing and biochemical properties. The enzyme production was evaluated by cultivating the isolated microorganisms in liquid-state bioprocess using wheat bran as carbon source. Two fungi (M1, and SE and one bacterium (I4 strains were found to be xylanase producer, and several were proven to be outstanding producers of microbial xylanase. The strains producing xylanase secreted variable amounts of starch-debranching enzymes and produced low level β-mannan-degrading enzyme systems. The bacterium strain was found to be capable of producing pectinolytic enzymes on wheat bran at high level. Some of the strains have good potential for use as sources of important industrial enzymes.

  3. COLONIZATION OF VIGNA RADIATA ROOTS BY CHROMIUM RESISTANT BACTERIAL STRAINS OF OCHROBACTRUM INTERMEDIUM, BACILLUS CEREUS AND BREVIBA CTERIUM SP.

    Institute of Scientific and Technical Information of China (English)

    MUHAMMAD Faisal; SHAHIDA Hasnain

    2005-01-01

    The present study deals with colonization potential of plant growth promoting bacterial strains ( Ochrobactrum intermedium, Bacillus cereus and Brevibacterium sp. ) on Vigna radiata roots. The roots were heavily colonized with O. intermedium and B. cereus as compared to Brevibacterium sp. O. intermedium mainly colonized rhizoplane while B. cereus occurred both on the rhizoplane and near root zone. O. intermedium and B. cereus were found to be present both on the rhizoplane and near root zone, while Brevibacterium only in the rhizosphere in the form of groups. The cells of B. cereus were found more in the sites where root exudates were existed. From the above results it was observed that the number of O. intermedium cells were large at root exudate site. Fig 2, Tab 1, Ref 15

  4. Partial Characterization of an Anti-Candida albicans Bacteriocin Produced by a Marine Strain of Bacillus sp., Sh10

    Directory of Open Access Journals (Sweden)

    Fatemeh Shayesteh

    2015-09-01

    Full Text Available The bacteriocin-producing strain Bacillus sp., Sh10, isolated from the marine environment, exhibited a broad spectrum of antimicrobial activity against different food spoilage and human pathogens, with a maximum inhibitory activity against Candida albicans. The inhibitory compound was sensitive to trypsin but resistant to proteinase K, lysozyme, lipase and &alpha-amylase. It was heat-stable and remained its activity after autoclaving. In addition, the antimicrobial substance demonstrated striking stability at low temperatures (4 and -20°C for up to one year and retained its activity in a wide pH range from 2 to 11. It was also stable and active in the presence of different surfactants, solvents and heavy metals. Analysis of the partially purified bacteriocin by SDS-PAGE showed an apparent molecular weight of ~11 KDa. This study reveals a remarkable potential of this bacteriocin to be used as a food preservative.

  5. Increasing the catalytic activity of Bilirubin oxidase from Bacillus pumilus: Importance of host strain and chaperones proteins.

    Science.gov (United States)

    Gounel, Sébastien; Rouhana, Jad; Stines-Chaumeil, Claire; Cadet, Marine; Mano, Nicolas

    2016-07-20

    Aggregation of recombinant proteins into inclusion bodies (IBs) is the main problem of the expression of multicopper oxidase in Escherichia coli. It is usually attributed to inefficient folding of proteins due to the lack of copper and/or unavailability of chaperone proteins. The general strategies reported to overcome this issue have been focused on increasing the intracellular copper concentration. Here we report a complementary method to optimize the expression in E. coli of a promising Bilirubin oxidase (BOD) isolated from Bacillus pumilus. First, as this BOD has a disulfide bridge, we switched E.coli strain from BL21 (DE3) to Origami B (DE3), known to promote the formation of disulfide bridges in the bacterial cytoplasm. In a second step, we investigate the effect of co-expression of chaperone proteins on the protein production and specific activity. Our strategy allowed increasing the final amount of enzyme by 858% and its catalytic rate constant by 83%.

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

    Science.gov (United States)

    Pueyo, Manuel T; Mutafci, Bruna A; Soto-Arriaza, Marco A; Di Mascio, Paolo; Carmona-Ribeiro, Ana M

    2014-01-01

    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.

  7. 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.

  8. Biodegradation of fluoranthene by a newly isolated strain of Bacillus stratosphericus from Mediterranean seawater of the Sfax fishing harbour, Tunisia.

    Science.gov (United States)

    Hentati, Dorra; Chebbi, Alif; Loukil, Slim; Kchaou, Sonia; Godon, Jean-Jacques; Sayadi, Sami; Chamkha, Mohamed

    2016-08-01

    A physico-chemical characterization of seawater taken from the fishing harbour of Sfax, Tunisia, revealed a contamination by organic and inorganic micropollutants. An aerobic marine halotolerant Bacillus stratosphericus strain FLU5 was isolated after enrichment on fluoranthene, a persistent and toxic polycyclic aromatic hydrocarbon (PAH). GC-MS analyses showed that strain FLU5 was capable of degrading almost 45 % of fluoranthene (100 mg l(-1)), without yeast extract added, after 30 days of incubation at 30 g l(-1) NaCl and 37 °C. In addition, the isolate FLU5 showed a remarkable capacity to grow on a wide range of aliphatic, aromatic and complex hydrocarbons. This strain could also synthesize a biosurfactant which was capable of reducing the surface tension of the cell-free medium, during the growth on fluoranthene. The biodegradative abilities of PAHs are promising and can be used to perform the bioremediation strategies of seawaters and marine sediments contaminated by hydrocarbons.

  9. Bacillus methylotrophicus Strain NKG-1, Isolated from Changbai Mountain, China, Has Potential Applications as a Biofertilizer or Biocontrol Agent

    Science.gov (United States)

    Ge, Beibei; Liu, Binghua; Nwet, Thinn Thinn; Zhao, Wenjun; Shi, Liming; Zhang, Kecheng

    2016-01-01

    Chemical pesticides are widely used in agriculture, which endangers both environmental health and food safety. Biocontrol is an environmentally-friendly and cost-effective green technique in environmental protection and agricultural production; it generally uses selected bioresources, including beneficial microorganisms. We isolated a novel bacterial strain (NKG-1) from the rare dormant volcanic soils of Changbai Mountain in China’s Jilin Province. The strain was identified as Bacillus methylotrophicus using morphological, biochemical, physiological, and phylogenetic 16S rDNA sequencing data. This strain was able to suppress mycelial growth and conidial germination of numerous plant pathogenic fungi on solid media. A greenhouse experiment showed that application of NKG-1 fermentation broth prior to inoculation of Botrytis cinerea, the cause of gray tomato mold, inhibited growth of the mold by 60%. Furthermore, application of a 100× dilution of NKG-1 fermentation broth to tomato seedlings yielded a significant increase in seedling fresh weight (27.4%), seedling length (12.5%), and root length (57.7%) compared to the control. When the same dosage was applied in the field, we observed increases in tomato plant height (14.7%), stem diameter (12.7%), crown width (16.3%), and maximum fruit diameter (11.5%). These results suggest that NKG-1 has potential commercial application as a biofertilizer or biocontrol agent. PMID:27832162

  10. Expression of cry1Ab gene from a novel Bacillus thuringiensis strain SY49-1 active on pest insects.

    Science.gov (United States)

    Azizoglu, Ugur; Ayvaz, Abdurrahman; Yılmaz, Semih; Karabörklü, Salih; Temizgul, Rıdvan

    2016-01-01

    In this study, the cry1Ab gene of previously characterized and Lepidoptera-, Diptera-, and Coleoptera-active Bacillus thuringiensis SY49-1 strain was cloned, expressed and individually tested on Ephestia kuehniella (Lepidoptera: Pyralidae) and Plodia interpunctella (Lepidoptera: Pyralidae) larvae. pET-cry1Ab plasmids were constructed by ligating the cry1Ab into pET28a (+) expression vector. Constructed plasmids were transferred to an Escherichia coli BL21 (DE3) strain rendered competent with CaCl2. Isopropyl β-d-1-thiogalactopyranoside was used to induce the expression of cry1Ab in E. coli BL21(DE3), and consequently, ∼130kDa of Cry1Ab was obtained. Bioassay results indicated that recombinant Cry1Ab at a dose of 1000μgg(-1) caused 40% and 64% mortality on P. interpunctella and E. kuehniella larvae, respectively. However, the mortality rates of Bt SY49-1 strains' spore-crystal mixture at the same dose were observed to be 70% on P. interpunctella and 90% on E. kuehniella larvae. The results indicated that cry1Ab may be considered as a good candidate in transgenic crop production and as an alternative biocontrol agent in controlling stored product moths.

  11. Identification, genetic diversity and cereulide producing ability of Bacillus cereus group strains isolated from Beninese traditional fermented food condiments.

    Science.gov (United States)

    Thorsen, Line; Azokpota, Paulin; Hansen, Bjarne Munk; Hounhouigan, D Joseph; Jakobsen, Mogens

    2010-08-15

    Bacillus cereus sensu lato is often detected in spontaneously fermented African foods but is rarely identified to species level. Only some of the B. cereus group species are reported to be pathogenic to humans and identification to species level is necessary to estimate the safety of these products. In the present study, a total of 19 Bacillus cereus group spp. isolated from afitin, iru and sonru, three spontaneously fermented African locust (Parkia biglobosa) bean based condiments produced in Benin, were investigated. The strains were isolated at 6, 12, 18, 24 and 48 h fermentation time. By using phenotypic and genotypic methods all of the isolates could be identified as B. cereus sensu stricto. The isolates were grouped according to their PM13 PCR (random amplification of polymorphic DNA PCR) fingerprint and formed two major clusters, one of which contained eight strains isolated from afitin (cluster 1). Highly similar PM13 profiles were obtained for seven of the isolates, one from afitin, one from iru and five from sonru (cluster 2). Four of the isolates, one from afitin and three from sonru, did not form any particular cluster. The PM13 profiles of cluster 2 isolates were identical to those which are specific to emetic toxin producers. Cereulide production of these isolates was confirmed by liquid chromatography mass spectrometry/mass spectrometry. This is the first report on cereulide producing B. cereus in African fermented foods. Occurrence of the opportunistic human pathogen B. cereus, which is able to produce emetic toxin in afitin, iru and sonru, could impose a health hazard. Interestingly, no reports on food poisoning from the consumption of the fermented condiments exist.

  12. Screening, Identification and Antibacterial Activities of Effective Thermotolerant Bacillus spp. Strains Isolated from Raw Milk

    Directory of Open Access Journals (Sweden)

    Kannikar SANTONG

    2008-01-01

    Full Text Available Forty-one isolates of Bacillus species were isolated from raw milk, analyzed using the spot on lawn and agar diffusion method in terms of their general inhibition effects to test bacteria (Escherichia coli TISTR 887 and Staphylococcus aureus TISTR 517. The results demonstrated that most isolates are effective against Gram-positive and Gram-negative bacteria whereas their extensive inhibition effect is particularly against Gram-positive bacteria. Only 2 effective thermotolerant isolates, BA8 and BA16, exerted broad spectrum antibacterial activities against both test bacteria.  Based on biochemical and physiological properties, they were classified as Brevibacillus laterosporus and Geobacillus thermoglucosidasius, respectively.

  13. A SIMPLE METHOD FOR ISOLATION OF SOME BACILLUS STRAINS WITH AN EXPRESSED ANTI-CANCER ACTIVITY

    Directory of Open Access Journals (Sweden)

    Francesco Marotta

    2006-02-01

    Full Text Available ABSTRACT:There is now increasing evidence that probiotic bacteria can provide health benefits to humans. In many areas of medicine (gastroenterology, urology, allergology, oncology and others, these sanative microorganisms may be considered as possible and viable alternatives applicable to patient care. Particularly, we have found that oral administration of Bacillus oligonitrophilus KU-1 cells can be used for treatment and prevention of some tumors. Here we present a simple method for isolation of bacteria with anticancer properties from soil.RESUMEN:Está aumentando la evidencia de que hay bacterias probióticas que pueden proporcionar beneficios saludables a los seres humanos. En muchas áreas de la medicina (gastroenterología, urología, alergología, oncología y otras, estos microorganismos pueden considerarse como alternativas posibles y viables aplicables al cuidado del paciente. Particularmente, nosotros hemos encontrado que la administración oral de células KU-1 Bacillus oligonitrophilus puede ser utilizada para el tratamiento y la prevención de algunos tumores. Aquí presentamos un método simple para aislamiento de suelos, de bacterias con características anticáncer.

  14. 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.

  15. Survival and conjugal transfer between Bacillus thuringiensis strains in aquatic environment Sobrevivência e conjugação de Bacillus thuringiensis em ambiente aquático

    OpenAIRE

    Luciana Furlaneto; Halha Ostrensky Saridakis; Olívia Márcia Nagy Arantes

    2000-01-01

    Field and laboratory studies were conducted to assess the survival of cells and spores and plasmid transfer between Bacillus thuringienis strains in aquatic environment. Results indicated that cells and spores of B. thuringiensis can survive for 10 days in water, without altering their number. The sporulation process began after 12-15 hours of inoculation of water. B. thuringiensis was able to transfer conjugative plasmids in the aquatic environment.O presente trabalho é um estudo sobre a sob...

  16. INFLUENCE OF BACILLUS AMYLOLIQUEFACIENS SUBSP. PLANTARUM IMV B-7404 STRAIN EXOMETABOLITES ON PHENYLALANINE AMMONIA-LYASE ACTIVITY IN WINTER WHEAT SEEDLINGS.

    Science.gov (United States)

    Dragovoz, I V; Korzh, Yu V; Leonova, N O; Iliash, V M; Avdeeva, L V

    2015-01-01

    Influence of Bacillus amyloliquefaciens subsp. plantarum IMV B-7404 strain exometabolites on phenylalanine ammonia-lyase (PAL, EC 4.3.1.24) activity in winter wheat seedlings has been studied. A significant increase of PAL activity at 4-6 hours after treatment of plant roots with Bacillus amyloliquefaciens subsp. plantarum IMVB-7404 strain exometabolites and in case of leaves infection with Bipolaris sorokiniana plant pathogen has been shown. It was established that PAL activity changed along with a decrease of the infected surface area of the leaves evidenced for the induction of response in winter wheat seedlings induced by IMV B-7404 strain exometabolites. It was concluded that the studied exponents could be used as model systems in the research of phytoimmunity induction mechanisms.

  17. 40 CFR 180.1243 - Bacillus subtilis var. amyloliquefaciens strain FZB24; exemption from the requirement of a...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bacillus subtilis var... EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From Tolerances § 180.1243 Bacillus subtilis... the requirement of a tolerance for residues of the Bacillus subtilis var. amyloliquefaciens...

  18. Biofilm inhibition and antimicrobial action of lipopeptide biosurfactant produced by heavy metal tolerant strain Bacillus cereus NK1.

    Science.gov (United States)

    Sriram, Muthu Irulappan; Kalishwaralal, Kalimuthu; Deepak, Venkataraman; Gracerosepat, Raja; Srisakthi, Kandasamy; Gurunathan, Sangiliyandi

    2011-07-01

    Biosurfactants are worthful microbial amphiphilic molecules with efficient surface-active and biological properties applicable to several industries and processes. Among them lipopeptides represent a class of microbial surfactants with increasing scientific, therapeutic and biotechnological interests. A heavy metal tolerant Bacillus strain has been isolated and the biofilm inhibition and antimicrobial activity of biosurfactant produced by the strain have been studied. Biosurfactant production was confirmed by the conventional screening methods including hemolytic activity, drop collapsing test, oil displacement test, emulsification and lipase production assays. The biosurfactant produced by this strain was a lipopeptide and exhibited strong surface activity. The biosurfactant has been characterized using FTIR, TLC and HPLC. The minimum active dose of this biosurfactant when compared with the other chemical surfactants was found as 0.150±0.06 μg. The critical micelle concentration was found to be 45 mg/l. The biosurfactant was found to be stable and active over a wide range of pH, temperature and NaCl concentration. It was also able to emulsify a wide range of hydrocarbons and oils thereby extending its application for the bioremediation of oil contaminated sites. The biosurfactant exhibited significant reduction in biofilm formation by pathogens and showed potent antimicrobial activity against various gram positive, gram negative bacteria and fungi. Agar diffusion assay for heavy metal resistance showed that the isolate was resistant to ferrous, lead and zinc. Considering the biofilm inhibition and antimicrobial property of biosurfactant, it can be utilized as a potential therapeutic molecule for numerous microbial infections. The heavy metal resistance of the strain can also be harnessed as an invaluable biological tool for in situ bioremediation.

  19. Detection of toxin proteins from Bacillus thuringiensis strain 4.0718 by strategy of 2D-LC-MS/MS.

    Science.gov (United States)

    Yang, Qi; Tang, Sijia; Rang, Jie; Zuo, Mingxing; Ding, Xuezhi; Sun, Yunjun; Feng, Pinghui; Xia, Liqiu

    2015-04-01

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

  20. Biodegradation of methyl red by Bacillus sp. strain UN2: decolorization capacity, metabolites characterization, and enzyme analysis.

    Science.gov (United States)

    Zhao, Ming; Sun, Peng-Fei; Du, Lin-Na; Wang, Guan; Jia, Xiao-Ming; Zhao, Yu-Hua

    2014-05-01

    Azo dyes are recalcitrant and refractory pollutants that constitute a significant menace to the environment. The present study is focused on exploring the capability of Bacillus sp. strain UN2 for application in methyl red (MR) degradation. Effects of physicochemical parameters (pH of medium, temperature, initial concentration of dye, and composition of the medium) were studied in detail. The suitable pH and temperature range for MR degradation by strain UN2 were respectively 7.0-9.0 and 30-40 °C, and the optimal pH value and temperature were respectively 8.0 and 35 °C. Mg(2+) and Mn(2+) (1 mM) were found to significantly accelerate the MR removal rate, while the enhancement by either Fe(3+) or Fe(2+) was slight. Under the optimal degradation conditions, strain UN2 exhibited greater than 98 % degradation of the toxic azo dye MR (100 ppm) within 30 min. Analysis of samples from decolorized culture flasks confirmed biodegradation of MR into two prime metabolites: N,N'dimethyl-p-phenyle-nediamine and 2-aminobenzoic acid. A study of the enzymes responsible for the biodegradation of MR, in the control and cells obtained during (10 min) and after (30 min) degradation, showed a significant increase in the activities of azoreductase, laccase, and NADH-DCIP reductase. Furthermore, a phytotoxicity analysis demonstrated that the germination inhibition was almost eliminated for both the plants Triticum aestivum and Sorghum bicolor by MR metabolites at 100 mg/L concentration, yet the germination inhibition of parent dye was significant. Consequently, the high efficiency of MR degradation enables this strain to be a potential candidate for bioremediation of wastewater containing MR.

  1. Combining Steinernema carpocapsae and Bacillus thuringienis strains for control of diamondback moth (Plutella xylostella).

    Science.gov (United States)

    Yi, X; Ehlers, R U

    2006-01-01

    The diamondback moth (DBM), Plutella xylostella, is a major pest of crucifers and has developed resistance against all chemical insecticides, even against the biological insecticide Bacillus thuringiensis (Bt). Entomopathogenic nematodes (EPN) as novel biological insecticide were successfully used against DBM. We investigated the potential of Bt to enhance the virulence of EPN and the possibility to reduce the application density by exploiting synergistic effects. The interaction between different combinations of Bt and EPN were tested against early 3rd instar P. xylostella on 2 cm2 cabbage leaf discs. All of the combinations had additive effects. Synergistic results were exceptional. The result indicate that a joint application of Bt and EPN is not recommended. But alternating applications are a measure to manage resistance development against Bt.

  2. Computational analysis of collagenase from different Vibrio, Clostridium and Bacillus strains to find new enzyme sources

    Directory of Open Access Journals (Sweden)

    Navid Nezafat

    2015-12-01

    Full Text Available Collagenase is one the important enzyme, which is applied in varied fields ranging from tannery, food and cosmetic industries to clinical therapies. Currently, the commercially available collagenase enzyme has been produced by Clostridium histolyticum bacteria. In our study, in order to find new sources of collagenase producer, 30 collagenases from different species of Clostridium, Vibrio and Bacillus were evaluated from the view of phylogenetic relation, domain architecture and physiochemical features. Totally our results indicate that the non-pathogenic C. novyi (NT with the aliphatic index (80.68, instability index (27, pI (6.54, Mw (112.838 kDa and two PPC domain could be suggested as a potent bacteria for industrial production of collagenase.

  3. Occurrence and Genetic Diversity of Bacillus anthracis Strains Isolated in an Active Wool-Cleaning Factory▿

    Science.gov (United States)

    Wattiau, Pierre; Klee, Silke R.; Fretin, David; Van Hessche, Mieke; Ménart, Marie; Franz, Tatjana; Chasseur, Camille; Butaye, Patrick; Imberechts, Hein

    2008-01-01

    Culturable microorganisms from various samples taken at an active factory performing wool and goat hair cleaning were isolated and analyzed. Bacillus anthracis was found in air filter dust, wastewater, and goat hairs, where it accounted for approximately 1% of the total counts of viable bacteria. Consistent with the countries of origin of the processed material (South Caucasian and Middle Eastern), all B. anthracis isolates belonged to the same phylogenetic cluster, as determined by variable-number tandem repeat (VNTR) typing at eight loci. Within this cluster, five closely related VNTR subtypes could be identified, of which two were previously unreported. Additional diversity was observed when more sensitive genetic markers were assayed, demonstrating the multifocal nature of goat hair contamination. Goat hair originating from areas where anthrax is endemic remains a material with high biological risk for modern woolworkers. PMID:18487406

  4. Growth energetics of an alkaline serine protease-producing strain of Bacillus clausii during continuous cultivation

    DEFF Research Database (Denmark)

    Christiansen, Torben; Nielsen, Jens

    2002-01-01

    the theoretical co-factor and building block requirements needed for biomass formation were calculated. Using the stoichiometric models and data for growth on glucose and citrate the amount of ATP needed for biomass synthesis was estimated to 42.0 mmol ATP/gDW, the P/O ratio to 0.68 and the ATP maintenance to 2......Glucose-limited chemostats were used to determine the growth yields of biomass of Bacillus clausii PP 473-8 producing an alkaline serine protease Savinase (Novozymes A/S, Bagsvaerd, Denmark) and a low yield of biomass on oxygen was observed. The energy metabolism was investigated further by setting...... up simple stoichiometric models for growth on glucose and citrate. In order to determine the parameters in the models, a macromolecular biomass composition was determined based on measured values of protein and RNA combined with literature data. From the macromolecular composition of the biomass...

  5. Complete Genome Sequence of Bacillus subtilis Strain 29R7-12, a Piezophilic Bacterium Isolated from Coal-Bearing Sediment 2.4 Kilometers below the Seafloor

    Science.gov (United States)

    Wei, Yuli; Cao, Junwei; Kato, Chiaki; Cui, Weicheng

    2017-01-01

    ABSTRACT Here, we report the genome sequence of Bacillus subtilis strain 29R7-12, a piezophilic bacterium isolated from coal-bearing sediment down to ~2.4 km below the ocean floor in the northwestern Pacific. The strain is a Gram-positive spore-forming bacterium, closely related to Bacillus subtilis within the phylum Firmicutes. This is the first complete genome sequence of a Bacillus subtilis strain from the deep biosphere. The genome sequence will provide a valuable resource for comparative studies of microorganisms from the surface and subsurface environments. PMID:28232436

  6. Parasporin-2 from a New Bacillus thuringiensis 4R2 Strain Induces Caspases Activation and Apoptosis in Human Cancer Cells

    Science.gov (United States)

    Asselin, Eric; Parent, Sophie; Côté, Jean-Charles; Sirois, Marc

    2015-01-01

    In previous studies, parasporin-2Aa1, originally isolated from Bacillus thuringiensis strain A1547, was shown to be cytotoxic against specific human cancer cells but the mechanisms of action were not studied. In the present study, we found that proteinase K activated parasporin-2Aa1 protein isolated from a novel B. thuringiensis strain, 4R2, was specifically cytotoxic to endometrial, colon, liver, cervix, breast and prostate cancer. It showed no toxicity against normal cells. Upon treatment with proteinase K-activated parasporin-2Aa1, morphological changes were observed and western blot analysis revealed the cleavage of poly (ADP-Ribose) polymerase, caspase-3 and caspase-9 in cancer cell lines exclusively, indicative of programmed cell death, apoptosis. Flow cytometry analyses,using propidium iodide and annexin V, as well as a caspases 3/7 assay confirmed apoptosis induction. Further analyses were performed to study survival pathways, including AKT, XIAP, ERK1/2 and PAR-4, a known inducer of apoptosis. These results indicate that parasporin-2Aa1 is a selective cytotoxic protein that induces apoptosis in various human cancer cell lines from diverse tissues. PMID:26263002

  7. An Antimicrobial Metabolite from Bacillus sp.: Significant activity against pathogenic bacteria including multidrug-resistant clinical strains

    Directory of Open Access Journals (Sweden)

    AJAY GHOSH CHALASANI

    2015-12-01

    Full Text Available In this study, the cell free modified trypticase soya broth (pH 7.4+0.2 of Bacillus subtilis URID 12.1 showed significant antimicrobial activity against multidrug-resistant strains of Staphylococcus aureus, S. epidermidis, Streptococcus pyogenes and Enterococcus faecalis. The partially purified antimicrobial molecule was found to be resistant to extremes of pH and temperatures and also to higher concentrations of trypsin and proteinase K. The antimicrobial molecule was purified by a three-step method that included reverse-phased high performance liquid chromatography (RP-HPLC. The minimum inhibitory concentration (MIC values were determined for 11 species of bacteria using a microbroth dilution technique. The HPLC-purified fraction showed the MICs ranging from 0.5 to 1 µg/ml for methicillin and vancomycin resistant Staphylococcus aureus (MVRSA and methicillin-resistant Staphylococcus epidermidis (MRSE strains. The molecular mass of the antimicrobial compound was determined to be 842.37 Da. The same antimicrobial fraction showed negligible haemolytic activity against human red blood cells even at a concentration as high as 100µg/ml. Because of its significant antimicrobial activity at low MIC values coupled with its non-haemolytic property, it may prove to be a novel antimicrobial lead molecule.

  8. EFEKTIVITAS Bacillus thuringiensis H-14 STRAIN LOKAL DALAM BUAH KELAPA TERHADAP LARVA Anopheles sp dan Culex sp di KAMPUNG LAUT KABUPATEN CILACAP

    Directory of Open Access Journals (Sweden)

    Blondine Ch. P

    2013-07-01

    Full Text Available Abstrak Bacillus thuringiensis serotipe H-14 strain lokal adalah bakteri patogen bersifat target spesifiknya larva nyamuk, aman bagi mamalia dan lingkungan. Penelitian bertujuan menentukan efektivitas B. thuringiensis H-14 strain lokal yang dikembangbiakkan dalam buah kelapa untuk pengendalian larva Anopheles sp dan Culex sp. Rancangan eksperimental semu, terdiri dari kelompok perlakuan dan kontrol. Bacillus thuringiensis H-14 strain lokal dikembangbiakan dalam10 buah kelapa umur 6–8 bulan, dengan berat kira-kira 1 kg, telah berisi air kelapa sekitar 400-500 ml/buah kelapa yang diperoleh dari Desa Klaces, Kampung Laut, Kabupaten Cilacap. Diinkubasi selama 14 hari pada temperatur kamar dan ditebarkan di 6 kolam yang menjadi habitat perkembangbiakan larva nyamuk dengan luas berkisar 3–100 m2.Hasil yang diperoleh menunjukkan efektivitas B. thuringiensis H-14 strain lokal terhadap larva Anopheles sp dan Culex sp selama 1 hari sesudah penebaran kematian larva berturut-turut sebesar 80–100% dan 79,31–100%. Sedangkan pada hari ke-14 sebesar 69,30–76,71% dan 67,69–86,04%. Buah kelapa dapat digunakan sebagai media lokal alternatif untuk pengembangbiakan B. thuringiensis H-14 strain lokal Kata kunci: B. thuringiensis H-14,  strain  lokal, buah kelapa, pengendalian larva Abstract Bacillus thuringiensis serotype H-14 local strain is pathogenic bacteria which specific  target to mosquito larvae. It is safe for mammals and enviroment. The aims of this study was to determine the effectivity of B. thuringiensis H-14 local strain which culturing in thecoconut wates against Anopheles sp and Culex sp mosquito larvae. This research is quasi experiment which consist of treated  and control groups. Bacillus thuringiensis H-14 local strain was cultured in 10 coconuts with 6–8 months age with weight around 1 kg that contained were approximately 400-500 ml/coconut were taken from Klaces village, Kampung Laut. After that the coconuts incubated for 14

  9. An attenuated strain of Bacillus anthracis (CDC 684 has a large chromosomal inversion and altered growth kinetics

    Directory of Open Access Journals (Sweden)

    Ivins Bruce E

    2011-09-01

    Full Text Available Abstract Background An isolate originally labeled Bacillus megaterium CDC 684 was found to contain both pXO1 and pXO2, was non-hemolytic, sensitive to gamma-phage, and produced both the protective antigen and the poly-D-glutamic acid capsule. These phenotypes prompted Ezzell et al., (J. Clin. Microbiol. 28:223 to reclassify this isolate to Bacillus anthracis in 1990. Results We demonstrate that despite these B. anthracis features, the isolate is severely attenuated in a guinea pig model. This prompted whole genome sequencing and closure. The comparative analysis of CDC 684 to other sequenced B. anthracis isolates and further analysis reveals: a CDC 684 is a close relative of a virulent strain, Vollum A0488; b CDC 684 defines a new B. anthracis lineage (at least 51 SNPs that includes 15 other isolates; c the genome of CDC 684 contains a large chromosomal inversion that spans 3.3 Mbp; d this inversion has caused a displacement of the usual spatial orientation of the origin of replication (ori to the termination of replication (ter from 180° in wild-type B. anthracis to 120° in CDC 684 and e this isolate also has altered growth kinetics in liquid media. Conclusions We propose two alternative hypotheses explaining the attenuated phenotype of this isolate. Hypothesis 1 suggests that the skewed ori/ter relationship in CDC 684 has altered its DNA replication and/or transcriptome processes resulting in altered growth kinetics and virulence capacity. Hypothesis 2 suggests that one or more of the single nucleotide polymorphisms in CDC 684 has altered the expression of a regulatory element or other genes necessary for virulence.

  10. Use of fatty acid methyl ester profiles for discrimination of Bacillus cereus T-strain spores grown on different media.

    Science.gov (United States)

    Ehrhardt, Christopher J; Chu, Vivian; Brown, TeeCie; Simmons, Terrie L; Swan, Brandon K; Bannan, Jason; Robertson, James M

    2010-03-01

    The goal of this study was to determine if cellular fatty acid methyl ester (FAME) profiling could be used to distinguish among spore samples from a single species (Bacillus cereus T strain) that were prepared on 10 different medium formulations. To analyze profile differences and identify FAME biomarkers diagnostic for the chemical constituents in each sporulation medium, a variety of statistical techniques were used, including nonmetric multidimensional scaling (nMDS), analysis of similarities (ANOSIM), and discriminant function analysis (DFA). The results showed that one FAME biomarker, oleic acid (18:1 omega9c), was exclusively associated with spores grown on Columbia agar supplemented with sheep blood and was indicative of blood supplements that were present in the sporulation medium. For spores grown in other formulations, multivariate comparisons across several FAME biomarkers were required to discern profile differences. Clustering patterns in nMDS plots and R values from ANOSIM revealed that dissimilarities among FAME profiles were most pronounced when spores grown with disparate sources of complex additives or protein supplements were compared (R > 0.8), although other factors also contributed to FAME differences. DFA indicated that differentiation could be maximized with a targeted subset of FAME variables, and the relative contributions of branched FAME biomarkers to group dissimilarities changed when different media were compared. When taken together, these analyses indicate that B. cereus spore samples grown in different media can be resolved with FAME profiling and that this may be a useful technique for providing intelligence about the production methods of Bacillus organisms in a forensic investigation.

  11. Genome-Guided Insights into the Plant Growth Promotion Capabilities of the Physiologically Versatile Bacillus aryabhattai Strain AB211

    Science.gov (United States)

    Bhattacharyya, Chandrima; Bakshi, Utpal; Mallick, Ivy; Mukherji, Shayantan; Bera, Biswajit; Ghosh, Abhrajyoti

    2017-01-01

    Bacillus aryabhattai AB211 is a plant growth promoting, Gram-positive firmicute, isolated from the rhizosphere of tea (Camellia sinensis), one of the oldest perennial crops and a major non-alcoholic beverage widely consumed all over the world. The whole genome of B. aryabhattai AB211 was sequenced, annotated and evaluated with special focus on genomic elements related to plant microbe interaction. It’s genome sequence reveals the presence of a 5,403,026 bp chromosome. A total of 5226 putative protein-coding sequences, 16 rRNA, 120 tRNA, 8 ncRNAs, 58 non-protein coding genes, and 11 prophage regions were identified. Genome sequence comparisons between strain AB211 and other related environmental strains of B. aryabhattai, identified about 3558 genes conserved among all B. aryabhattai genomes analyzed. Most of the common genes involved in plant growth promotion activities were found to be present within core genes of all the genomes used for comparison, illustrating possible common plant growth promoting traits shared among all the strains of B. aryabhattai. Besides the core genes, some genes were exclusively identified in the genome of strain AB211. Functional annotation of the genes predicted in the strain AB211 revealed the presence of genes responsible for mineral phosphate solubilization, siderophores, acetoin, butanediol, exopolysaccharides, flagella biosynthesis, surface attachment/biofilm formation, and indole acetic acid production, most of which were experimentally verified in the present study. Genome analysis and experimental evidence suggested that AB211 has robust central carbohydrate metabolism implying that this bacterium can efficiently utilize the root exudates and other organic materials as an energy source. Genes for the production of peroxidases, catalases, and superoxide dismutases, that confer resistance to oxidative stresses in plants were identified in AB211 genome. Besides these, genes for heat shock tolerance, cold shock tolerance

  12. Screening and characterization of extracelluar L-asparaginase producing Bacillus subtilis strain hswx88, isolated from Taptapani hotspring of Odisha, India

    Institute of Scientific and Technical Information of China (English)

    Biswaprakash Pradhan; Sashi K Dash; Sabuj Sahoo

    2013-01-01

    Objective: To screen and isolate an eco-friendly, a thermophilic and potent L-asparaginase producing bacterium, with novel immunological properties that may obviates hypersensitivity reactions. Methods: In the present study bacterial strain isolated for extracellular L-asparaginase production from hotspring, identified by morphological, biochemical and physiological tests followed by 16S rDNA technology and the L-asparaginase production ability was tested by both semi quantitative and quantitative enzymatic assay. Result:The bacterial strain was identified as Bacillus subtilis strain hswx88 (GenBank Accession Number: JQ237656.1). The extracellular enzyme yielding capacity isolate Bacillus subtilis strain hswx88 (23.8 IU/mL) was found to be 1.7 and 14.5 times higher than the reference organism Pectobacterium carotovorum MTCC 1428 (14.2 IU/mL) and Bacillus sp. BCCS 034 (1.64 IU/mL). Conclusion: The isolate is eco-friendly and useful to produce bulk quantity of extracellular, thermophilic L-asparaginase for the treatment of various tumor cases and for preparation of acrylamide free fry food preparation.

  13. The presence of sboA and spaS genes and antimicrobial peptides subtilosin A and subtilin among Bacillus strains of the Amazon basin.

    Science.gov (United States)

    Velho, Renata Voltolini; Basso, Ana Paula; Segalin, Jeferson; Costa-Medina, Luis Fernando; Brandelli, Adriano

    2013-03-01

    This report demonstrates the usefulness of PCR for the genes spaS and sboA as a means of identifying Bacillus strains with a potential to produce subtilin and subtilosin A. One collection strain and five Bacillus spp. isolated from aquatic environments in the Amazon basin were screened by PCR using primers for sboA and spaS designed specifically for this study. The sequences of the PCR products showed elevated homology with previously described spaS and sboA genes. Antimicrobial peptides were isolated from culture supernatants and analyzed by mass spectrometry. For all samples, the mass spectra revealed clusters with peaks at m/z 3300-3500 Da, corresponding to subtilosin A, subtilin and isoforms of these peptides. These results suggest that the antimicrobial activity of these strains may be associated with the production of subtilosin A and/or subtilin. The PCR used here was efficient in identifying novel Bacillus strains with the essential genes for producing subtilosin A and subtilin.

  14. The presence of sboA and spaS genes and antimicrobial peptides subtilosin A and subtilin among Bacillus strains of the Amazon basin

    Science.gov (United States)

    Velho, Renata Voltolini; Basso, Ana Paula; Segalin, Jeferson; Costa-Medina, Luis Fernando; Brandelli, Adriano

    2013-01-01

    This report demonstrates the usefulness of PCR for the genes spaS and sboA as a means of identifying Bacillus strains with a potential to produce subtilin and subtilosin A. One collection strain and five Bacillus spp. isolated from aquatic environments in the Amazon basin were screened by PCR using primers for sboA and spaS designed specifically for this study. The sequences of the PCR products showed elevated homology with previously described spaS and sboA genes. Antimicrobial peptides were isolated from culture supernatants and analyzed by mass spectrometry. For all samples, the mass spectra revealed clusters with peaks at m/z 3300–3500 Da, corresponding to subtilosin A, subtilin and isoforms of these peptides. These results suggest that the antimicrobial activity of these strains may be associated with the production of subtilosin A and/or subtilin. The PCR used here was efficient in identifying novel Bacillus strains with the essential genes for producing subtilosin A and subtilin. PMID:23569414

  15. Surfactin analogues produced by Bacillus subtilis strains grown on rapeseed cake

    Science.gov (United States)

    Jajor, Paweł; Piłakowska-Pietras, Dorota; Krasowska, Anna; Łukaszewicz, Marcin

    2016-12-01

    Microbiologically produced surface acting compounds (biosurfactants) have very interesting properties with many potential industrial applications. Lipopeptides is a particularly promising group of biosurfactants in respect to the potentially huge number of various chemical structures. The chemical diversity results from fatty acid moiety (e.g. length, saturation, branching or hydroxylation) and type and sequence of the amino acids in the peptide chain. The limiting factor for the design and analysis of various lipopeptides is the ability of the targeted biosynthesis. Biosynthesis of particular lipopeptides may be potentially achieved by strain selection, culture conditions, or molecular engineering. The well-known lipopeptedes (surfactins, iturins, and fengycins) producer is B. subtilis. The aim of this study was to study targeted surfactin structural analogues biosynthesis in response to culture conditions in view of the design and production of tailor-made lipopeptides. Two B. subtilis strains (KB1 and #309) were tested for surfactin production. Both strains produced a mixture of five major surfactin analogues with the number of carbons in an alkyl chain ranging from 12 to 16. The two strains differed with respect to their oxygen demand for optimal surfactin biosynthesis (lower oxygen demand for KB1). The amount of air influenced the relative ratios of surfactin analogues. Lower oxygen amount decreased the share of C15 analogues while it increased the share of C12 analogues. Thus, the biosynthesis of a desired surfactin analogue may controlled by both strain and culture conditions.

  16. Characterization and evaluation of Bacillus amyloliquefaciens strain WF02 regarding its biocontrol activities and genetic responses against bacterial wilt in two different resistant tomato cultivars.

    Science.gov (United States)

    Huang, Chu-Ning; Lin, Chan-Pin; Hsieh, Feng-Chia; Lee, Sook-Kuan; Cheng, Kuan-Chen; Liu, Chi-Te

    2016-11-01

    Bacillus amyloliquefaciens strain WF02, isolated from soil collected at Wufeng Mountain, Taiwan, has siderophore-producing ability and in vitro antagonistic activity against bacterial wilt pathogen. To determine the impact of plant genotype on biocontrol effectiveness, we treated soil with this strain before infecting susceptible (L390) and moderately resistant (Micro-Tom) tomato cultivars with Ralstonia solanacearum strain Pss4. We also compared the efficacy of this strain with that of commercial Bacillus subtilis strain Y1336. Strain WF02 provided longer lasting protection against R. solanacearum than did strain Y1336 and controlled the development of wilt in both cultivars. To elucidate the genetic responses in these plants under WF02 treatment, we analyzed the temporal expression of defense-related genes in leaves. The salicylic acid pathway-related genes phenylalanine ammonia-lyase and pathogenesis-related protein 1a were up-regulated in both cultivars, whereas expression of the jasmonic acid pathway-related gene lipoxygenase was only elevated in the susceptible tomato cultivar (L390). These results suggest that WF02 can provide protection against bacterial wilt in tomato cultivars with different levels of disease resistance via direct and indirect modes of action.

  17. 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)

  18. Characterization and genomic analysis of chromate resistant and reducing Bacillus cereus strain SJ1

    Directory of Open Access Journals (Sweden)

    He Minyan

    2010-08-01

    Full Text Available Abstract Background Chromium is a toxic heavy metal, which primarily exists in two inorganic forms, Cr(VI and Cr(III. Chromate [Cr(VI] is carcinogenic, mutational, and teratogenic due to its strong oxidizing nature. Biotransformation of Cr(VI to less-toxic Cr(III by chromate-resistant and reducing bacteria has offered an ecological and economical option for chromate detoxification and bioremediation. However, knowledge of the genetic determinants for chromate resistance and reduction has been limited so far. Our main aim was to investigate chromate resistance and reduction by Bacillus cereus SJ1, and to further study the underlying mechanisms at the molecular level using the obtained genome sequence. Results Bacillus cereus SJ1 isolated from chromium-contaminated wastewater of a metal electroplating factory displayed high Cr(VI resistance with a minimal inhibitory concentration (MIC of 30 mM when induced with Cr(VI. A complete bacterial reduction of 1 mM Cr(VI was achieved within 57 h. By genome sequence analysis, a putative chromate transport operon, chrIA1, and two additional chrA genes encoding putative chromate transporters that likely confer chromate resistance were identified. Furthermore, we also found an azoreductase gene azoR and four nitroreductase genes nitR possibly involved in chromate reduction. Using reverse transcription PCR (RT-PCR technology, it was shown that expression of adjacent genes chrA1 and chrI was induced in response to Cr(VI but expression of the other two chromate transporter genes chrA2 and chrA3 was constitutive. In contrast, chromate reduction was constitutive in both phenotypic and gene expression analyses. The presence of a resolvase gene upstream of chrIA1, an arsenic resistance operon and a gene encoding Tn7-like transposition proteins ABBCCCD downstream of chrIA1 in B. cereus SJ1 implied the possibility of recent horizontal gene transfer. Conclusion Our results indicate that expression of the chromate

  19. Glyphosate biodegradation and potential soil bioremediation by Bacillus subtilis strain Bs-15.

    Science.gov (United States)

    Yu, X M; Yu, T; Yin, G H; Dong, Q L; An, M; Wang, H R; Ai, C X

    2015-11-23

    Glyphosate and glyphosate-containing herbicides have an adverse effect on mammals, humans, and soil microbial ecosystems. Therefore, it is important to develop methods for enhancing glyphosate degradation in soil through bioremediation. We investigated the potential of glyphosate degradation and bioremediation in soil by Bacillus subtilis Bs-15. Bs-15 grew well at high concentrations of glyphosate; the maximum concentration tolerated by Bs-15 reached 40,000 mg/L. The optimal conditions for bacterial growth and glyphosate degradation were less than 10,000 mg/L glyphosate, with a temperature of 35°C and a pH of 8.0. Optimal fermentation occurred at 180 rpm for 60 h with an inoculum ratio of 4%. Bs-15 degraded 17.65% (12 h) to 66.97% (96 h) of glyphosate in sterile soil and 19.01% (12 h) to 71.57% (96 h) in unsterilized soil. Using a BIOLOG ECO plate test, we observed no significant difference in average well color development values between the soil inoculated with Bs-15 and the control soil before 72 h, although there was a significant difference (P glyphosate-containing herbicides, increasing the microbial functional diversity in glyphosate-contaminated soils and thus enhancing the bioremediation of glyphosate-contaminated soils.

  20. 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 free...... amino acids (FAA) during the fermentation of ALBP by the Bacillus isolates were studied by SDS-PAGE and cation exchange chromatography. The profile of soluble proteins changed with the fermentation time and varied depending on the isolate. The quantity of total FAA and essential FAA such as lysine...... was increased sharply between 24 and 48 h of fermentation and differed among the isolates. Simultaneously, a pH increase was observed. Cysteine, methionine, leucine, isoleucine, tyrosine and phenylalaline appeared during fermentation.Conclusion: The Bacillus isolates studied degraded ALBP leading to a profile...

  1. Complete genome sequence of thermophilic Bacillus smithii type strain DSM 4216T

    DEFF Research Database (Denmark)

    Bosma, Elleke Fenna; Koehorst, Jasper J.; van Hijum, Sacha A. F. T.

    2016-01-01

    determined the complete genomic sequence of the B. smithii type strain DSM 4216T, which consists of a 3,368,778 bp chromosome (GenBank accession number CP012024.1) and a 12,514 bp plasmid (GenBank accession number CP012025.1), together encoding 3880 genes. Genome annotation via RAST was complemented...

  2. Characterization of volatiles of necrotic Stenocereus thurberi and Opuntia littoralis and toxicity and olfactory preference of Drosophila melanogster, D. mojavensis wrigleyi, and D. mojavensis sonorensis to necrotic cactus volatiles.

    Science.gov (United States)

    Wright, Cynthia R; Setzer, William N

    2014-08-01

    Drosophila mojavensis wrigleyi and D. mojavensis sonorensis are geographically separated races of cactophilic fruit flies. D. mojavensis sonorensis inhabits the Sonoran Desert and utilizes necrotic rots of Stenocereus thurberi Engelm. as a food source and to oviposit while D. mojavensis wrigleyi inhabits Santa Catalina Island, California and utilizes the necrotic rots of Opuntia littoralis (Engelm.) Cockerell. The objectives of this study were to determine the volatile compositions of the necrotic cacti and to determine if the volatile components show either selective toxicity or attraction toward the fruit flies. The volatile chemical compositions of field-rot specimens of both necrotic cacti were obtained by dynamic headspace (purge-and-trap) and hydrodistillation techniques and analyzed by gas chromatography - mass spectrometry. The volatile fraction of necrotic S. thurberi early rot was dominated by carboxylic acids (84.8%) and the late rot by p-cresol (32.6% in the dynamic headspace sample and 55.9% in the hydrodistilled sample). O. littoralis volatiles were dominated by carboxylic acids (86% in the dynamic headspace sample and 89.1% in the hydrodistilled sample). Fifteen compounds that were identified in the necrotic rot volatiles were used to test insecticidal activity and olfactory preference on the cactophilic Drosophila species, as well as D. melanogaster. Differences in toxicity and olfactory preference were observed between the different taxa. Both races of D. mojavensis exhibited toxicity to benzaldehyde and 2-nonanone, while butanoic acid and palmitic acid were tolerated at high concentrations. D. m. wrigleyi demonstrated a greater olfactory preference for anisole, butanoic acid, 2-heptanone, and palmitic acid than did D. m. sonorensis, while D. m. sonorensis demonstrated a greater preference for hexadecane, octanoic acid, and oleic acid than did D. m. wrigleyi.

  3. Synergism between wild-type Bacillus thuringiensis subsp. israelensis and B. sphaericus strains: a study based on isobolographic analysis and histopathology.

    Science.gov (United States)

    Sreshty, M Asha Latha; Kumar, K Pranay; Murty, U S N

    2011-04-01

    Prevention is the best resistance management strategy in integrated vector control programs. Combined use of insecticides of different classes that interact synergistically and show multi-site actions within the insect is recognized as a potential key strategy to be implemented even before the onset of resistance. The present study is aimed at harvesting the benefits of synergism between the wild-type Bacillus thuringiensis subsp. israelensis-H14 (Bti) and Bacillus sphaericus-2362 (Bs) strains by evaluating six different combinations of mixtures toxic to Aedes and Culex mosquito larvae. Isobolographic analysis was performed to distinguish the synergistic combinations of Bti and Bs, followed by determination of the degree of synergism through synergy and improvement factors. Furthermore, the speed of activity of Bs when combined with Bti is studied by histopathological investigations on the fate of midgut muscles of mosquito larvae upon exposure to individual wild-type strains as well as their mixtures.

  4. Isolation of Bacillus sp. strains capable of decomposing alkali lignin and their application in combination with lactic acid bacteria for enhancing cellulase performance.

    Science.gov (United States)

    Chang, Young-Cheol; Choi, Dubok; Takamizawa, Kazuhiro; Kikuchi, Shintaro

    2014-01-01

    Effective biological pretreatment method for enhancing cellulase performance was investigated. Two alkali lignin-degrading bacteria were isolated from forest soils in Japan and named CS-1 and CS-2. 16S rDNA sequence analysis indicated that CS-1 and CS-2 were Bacillus sp. Strains CS-1 and CS-2 displayed alkali lignin degradation capability. With initial concentrations of 0.05-2.0 g L(-1), at least 61% alkali lignin could be degraded within 48 h. High laccase activities were observed in crude enzyme extracts from the isolated strains. This result indicated that alkali lignin degradation was correlated with laccase activities. Judging from the net yields of sugars after enzymatic hydrolysis, the most effective pretreatment method for enhancing cellulase performance was a two-step processing procedure (pretreatment using Bacillus sp. CS-1 followed by lactic acid bacteria) at 68.6%. These results suggest that the two-step pretreatment procedure is effective at accelerating cellulase performance.

  5. Cloning and expression of the fusion protein of interleukin-2 and ESAT6 in Mycobacterium bovis Bacillus Calmette Guérin strain

    Institute of Scientific and Technical Information of China (English)

    FAN Xiong-lin; WANG Li-mei; LU Xian-yu; TU Zhi-guang; SHI Chang-hong; XU Zhi-kai

    2005-01-01

    @@ Tuberculosis (TB) is the most common cause of death in infectious diseases; it is estimated that approximately 2 million people per year die of TB. The present available TB vaccine is a live attenuated strain, Mycobacterium bovis Bacillus Calmette Guérin (BCG). However, it has been shown that BCG has variable protective efficacy, ranging from 0 to 85% in different clinical experiments.1 Therefore, a new TB vaccine is urgently needed.

  6. Comparative Evaluation of Agroindustrial Byproducts for the Production of Alkaline Protease by Wild and Mutant Strains of Bacillus subtilis in Submerged and Solid State Fermentation

    OpenAIRE

    Hamid Mukhtar; Ikramul Haq

    2013-01-01

    The present study describes the screening of different agroindustrial byproducts for enhanced production of alkaline protease by a wild and EMS induced mutant strain of Bacillus subtilis IH-72EMS8. During submerged fermentation, different agro-industrial byproducts were tested which include defatted seed meals of rape, guar, sunflower, gluten, cotton, soybean, and gram. In addition to these meals, rice bran, wheat bran, and wheat flour were also evaluated for protease production. Of all the b...

  7. Characterization of the synergistic interaction between Beauveria bassiana strain GHA and Bacillus thuringiensis morrisoni strain tenebrionis applied against Colorado potato beetle larvae.

    Science.gov (United States)

    Wraight, S P; Ramos, M E

    2017-01-17

    Studies were undertaken to further characterize the previously identified synergistic activity of Bacillus thuringiensis- and Beauveria bassiana-based biopesticides against Colorado potato beetle (CPB). A flowable concentrate of B. thuringiensis morrisoni strain tenebrionis (Bt) (Novodor® FC) and a wettable powder of B. bassiana strain GHA (Bb) (Mycotrol® 22WP) were applied against CPB larval populations infesting potato in field plots. Novodor FC and an oil-dispersion formulation of Bb (Mycotrol ES) were applied against second-instar CPB larvae on potted potato plants in greenhouse tests under low relative humidity (RH), variable-temperature conditions. Each pathogen was applied alone and in combination (tank-mixed) with the other pathogen. In the field tests, each biopesticide was also combined with the spray-carrier (formulation without active ingredient) of the other pathogen. Results from the greenhouse tests showed that under warm, dry conditions, low activity of Mycotrol was counterbalanced by high activity of the Novodor, and under cool, somewhat more humid conditions, low Novodor activity was balanced by high activity of Mycotrol, with the result being a constant level of synergism (CPB mortality ca. 20 percentage points higher than predicted by independent action). Similar levels of synergism were observed under the markedly different conditions of the field and greenhouse environments, and the synergism was confirmed as arising from interaction of the two micobes, as the Bt spray carrier had no significant effect on efficacy of the Mycotrol product and the Bb spray carrier had no effect on the efficacy of Novodor. The great capacity of these two control agents to act in concert to control CPB is well documented (the fast-acting, toxic Bt acting to protect potato crops from defoliation and the slow-acting Bb reducing survival to the adult stage). These finding further underscore the strong complementary action of these agents applied jointly against CPB.

  8. Mutagenesis of the Bacillus edaphicus Strain NBT and Its Effect on Growth of Chili and Cotton

    Institute of Scientific and Technical Information of China (English)

    SHENG Xia-fang; XIA Juan-juan; CHEN Jue

    2003-01-01

    A strain NBT capable of dissolving silicate minerals and promoting plant growth was treatedwith UV+LiCl. Thirty-two mutants tolerable to 2% NaCl solution were obtained. However, through the sur-vival experiments in high osmatic pressure, high temperature and different acidities, two mutants of NBT-6and NBT-19 were finally obtained. They could survive from 10 % NaCl solution and tolerate 55℃, acidic (pH4) and alkalic (pH 10) conditions. The mutants had the same ability to release K from silicate minerals as thestarting strain NBT. Pot experiments with chili and cotton showed that both the mutants developed in the rhi-zosphere soils. The available P and K contents in the rhizosphere soils and plant biomass increased through in-oculating these bacteria.

  9. Microbial dynamics during shelf-life of industrial Ricotta cheese and identification of a Bacillus strain as a cause of a pink discolouration.

    Science.gov (United States)

    Sattin, E; Andreani, N A; Carraro, L; Fasolato, L; Balzan, S; Novelli, E; Squartini, A; Telatin, A; Simionati, B; Cardazzo, B

    2016-08-01

    Dairy products are perishable and have to be preserved from spoilage during the food chain to achieve the desired shelf-life. Ricotta is a typical Italian soft dairy food produced by heat coagulation of whey proteins and is considered to be a light and healthy product. The shelf-life of Ricotta could be extended, as required by the international food trade market; however, heat resistant microflora causes spoilage and poses issues regarding the safety of the product. Next-generation sequencing (NGS) applied to the Ricotta samples defined the composition of the microbial community in-depth during the shelf-life. The analysis demonstrated the predominance of spore-forming bacteria throughout the shelf-life, mostly belonging to Bacillus, Paenibacillus and Clostridium genera. A strain involved in spoilage and causing a pink discolouration of Ricotta was isolated and characterised as Bacillus mycoides/weihenstephanensis. This is the first report of a food discolouration caused by a toxigenic strain belonging to the Bacillus cereus group that resulted the predominant strain in the community of the defective ricotta. These results suggest that the processing of raw materials to eliminate spores and residual microflora could be essential for improving the quality and the safety of the product and to extend the shelf-life of industrial Ricotta.

  10. The mechanism of uranium transformation from U(VI) into nano-uramphite by two indigenous Bacillus thuringiensis strains.

    Science.gov (United States)

    Pan, Xiaohong; Chen, Zhi; Chen, Fanbing; Cheng, Yangjian; Lin, Zhang; Guan, Xiong

    2015-10-30

    The mechanism of uranium transformation from U(VI) into nano-uramphite by two indigenous Bacillus thuringiensis strains was investigated in the present work. Our data showed that the bacteria isolated from uranium mine possessed highly accumulation ability to U(VI), and the maximum accumulation capacity was around 400 mg U/g biomass (dry weight). X-ray powder diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FT-IR) analyzes indicated that the U(VI) was adsorbed on the bacterial surface firstly through coordinating with phosphate, CH2 and amide groups, and then needle-like amorphous uranium compounds were formed. With the extension of time, the extracellular crystalline substances were disappeared, but some particles were appeared in the intracellular region, and these particles were characterized as tetragonal-uramphite. Moreover, the disrupted experiment indicated that the cell-free extracts had better uranium-immobilization ability than cell debris. Our findings provided the understanding of the uranium transformation process from amorphous uranium to crystalline uramphite, which would be useful in the regulation of uranium immobilization process.

  11. Microscopic analysis of a native Bacillus thuringiensis strain from Malaysia that produces exosporium-enclosed parasporal inclusion.

    Science.gov (United States)

    Chai, Pui Fun; Rathinam, Xavier; Solayappan, Maheswaran; Ahmad Ghazali, Amir Hamzah; Subramaniam, Sreeramanan

    2014-10-01

    The current study focused on the microscopic studies of a native Bacillus thuringiensis strain isolated from Malaysia, Bt-S84-13a, that produced an unusual crystal type. Primary detection of parasporal inclusions using a phase contrast microscope presented one to two small crystal proteins in the sporulating cells of Bt-S84-13a. Compound light microscopic examination of autolysed Bt-S84-13a cells stained with 0.133% Coomassie Brilliant Blue showed two types of crystal morphology: small crystals independent of spores and spore-associated crystals. Surface structure analysis with a scanning electron microscope revealed spherical-like, coarse and wrinkled-looking crystal in Bt-S84-13a. A close-up observation of the crystal morphology using a transmission electron microscope also demonstrated two parasporal inclusions in Bt-S84-13a. One inclusion was deposited against the forespore and was in a shape of incomplete rectangular. Another smaller inclusion was developed within the exosporium and was rectangular in shape. However, the latter inclusion was found lack in another bacterial cell which was still in the early stages of sporulation. This unique crystal morphology may imply some biological potential in Bt-S84-13a.

  12. Characterization of Extracellular Penicilin G Acylase Produced by A New Local Strain of Bacillus subtilis BAC4

    Directory of Open Access Journals (Sweden)

    SUPARTONO

    2008-06-01

    Full Text Available Penicillin G acylase (PGA which catalyses penicillin G hydrolysis reaction is a key enzyme for the industrial production of penicilin G derivatives used in therapeutics. A new local strain of Bacillus subtilis BAC4 was found capable of producing extracellular PGA. However, characteristics of this extracellular PGA are not known. The goal of this research was to characterize the extracellular PGA produced by B. subtilis BAC4. Enzyme production was carried out by batch fermentation, followed by enzyme purification and characterization of the PGA. The PGA activity was determined by the Kornfeld method, with optimal activity for hydrolysing penicillin G observed at 43 oC and pH 8.5. The activation energy of penicillin G hydrolysis by the PGA of B. subtilis BAC4 was determined as 4.9 kcal.mol-1 and Vmax and Km values were found to be 0.7 µmole.min-1.mg-1 and 3.5 mM respectively. PGA catalytic activity was competitively inhibited by phenylacetic acid with an inhibition constant, Ki(PAA, of 347.2 mM. It was concluded that the extracellular PGA of B. subtilis BAC4 can hydrolyse penicillin G efficiently.

  13. Bioaugmenting anaerobic digestion of biosolids with selected strains of Bacillus, Pseudomonas, and Actinomycetes species for increased methanogenesis and odor control.

    Science.gov (United States)

    Duran, Metin; Tepe, Nalan; Yurtsever, Deniz; Punzi, Vito L; Bruno, Charles; Mehta, Raj J

    2006-12-01

    The objective of this study was to evaluate the effects of bioaugmenting anaerobic biosolids digestion with a commercial product containing selected strains of bacteria from genera Bacillus, Pseudomonas, and Actinomycetes, along with ancillary organic compounds containing various micronutrients. Specifically, the effects of the bioaugment in terms of volatile solids destruction and generation and fate of odor-causing compounds during anaerobic digestion and during storage of the digested biosolids were studied. Two bench-scale anaerobic digesters receiving primary and secondary clarifier biosolids from various full-scale biological wastewater treatment plants were operated. One of the digesters received the bioaugment developed by Organica Biotech, while the other was operated as control. The bioaugmented digester generated 29% more net CH(4) during the 8 weeks of operation. In addition, the average residual propionic acid concentration in the bioaugmented digester was 54% of that in the control. The monitoring of two organic sulfide compounds, methyl mercaptan (CH(3)SH) and dimethyl sulfide (CH(3)SCH(3)), clearly demonstrated the beneficial effects of the bioaugmentation in terms of odor control. The biosolids digested in the bioaugmented digester generated a negligible amount of CH(3)SH during 10 days of post-digestion storage, while CH(3)SH concentration in the control reached nearly 300 ppm(v) during the same period. Similarly, peak CH(3)SCH(3) generated by stored biosolids from the bioaugmented digester was only 37% of that from the control.

  14. Tolerance and removal of chromium(Ⅵ) by Bacillus sp. strain YB-1 isolated from electroplating sludge

    Institute of Scientific and Technical Information of China (English)

    LIU Yun-guo; FENG Bao-ying; FAN Ting; ZHOU Hai-zhou; LI Xin

    2008-01-01

    Four chromium(Ⅵ)-resistant bacteria named YB-1, YB-2, YB-3 and YB-4 were isolated from Cr-electroplating sludge. YB-1 and YB-2 were identified as a member of Bacillus sp. based on morphology and Biolog Microstation System. The strain of YB-1 was selected to test for its resistance and ability to remove Cr(Ⅵ) from aqueous solution. The results indicate that YB-1 exhibits high MIC value which can almost reach 140 mg/L and the growth of YB-1 in liquid medium containing 60 mg/L Cr(Ⅵ) is affected especially in the late exponential phase and stationary phase. Furthermore, the potential of living and freeze-dried YB-1 biomass to remove Cr(Ⅵ) was studied in different pH, biosorbent dose, contact time and initial concentration using the batch method. At the optimal conditions, living and freeze-dried biomass are capable of absorbing 34.5 mg/g and 17.8 mg/g chromium(Ⅵ) at initial concentration of 60 mg/L, respectively. The adsorption data were fitted to Langmuir isotherm model for these two sorbents. Kinetic studies show that the rates of sorption all follow the pseudo-second order kinetics.

  15. Contemporaneous Production of Amylase and Protease through CCD Response Surface Methodology by Newly Isolated Bacillus megaterium Strain B69

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    Rajshree Saxena

    2014-01-01

    Full Text Available The enormous increase in world population has resulted in generation of million tons of agricultural wastes. Biotechnological process for production of green chemicals, namely, enzymes, provides the best utilization of these otherwise unutilized wastes. The present study elaborates concomitant production of protease and amylase in solid state fermentation (SSF by a newly isolated Bacillus megaterium B69, using agroindustrial wastes. Two-level statistical model employing Plackett-Burman and response surface methodology was designed for optimization of various physicochemical conditions affecting the production of two enzymes concomitantly. The studies revealed that the new strain concomitantly produced 1242 U/g of protease and 1666.6 U/g of amylase by best utilizing mustard oilseed cake as the substrate at 20% substrate concentration and 45% moisture content after 84 h of incubation. An increase of 2.95- and 2.04-fold from basal media was observed in protease and amylase production, respectively. ANOVA of both the design models showed high accuracy of the polynomial model with significant similarities between the predicted and the observed results. The model stood accurate at the bench level validation, suggesting that the design model could be used for multienzyme production at mass scale.

  16. Production, purification and characterization of thermostable α-amylase from soil isolate Bacillus sp. strain B-10

    Directory of Open Access Journals (Sweden)

    Ravindra Nath Singh

    2016-04-01

    Full Text Available A bacterial strain B-10 that produces α-amylase was isolated from compost and kitchen waste receiving agricultural soil. Based on microbiological and biochemical tests the isolate B-10 was identified as Bacillus sp. Alpha-amylase produced by this isolate was purified by (NH42SO4 precipitation and DEAE cellulose ion-exchange chromatography showing 15.91 and 48.21 fold purification, respectively. SDS-PAGE of the purified enzyme confirmed the purification and monomeric nature of the enzyme. The purified α-amylase showed maximum activity at pH 7 and temperature 50°C. The enzyme was significantly active in the temperature range of 30-60°C for the studied period of 2 h. During the incubation of purified enzyme at pH ranging from 5 to 10 for 24 h the maximum stability was observed at pH 7 followed by pH 8, whereas at extreme pH, the stability was very poor. Km and Vmax were found to be 1.4 mg/mL and 6.2 U/mL, respectively.

  17. 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.; van Beilen, J.; Caspers, M.; O'Brien, A.; de Koster, C.; Oomes, S.; Smelt, J.; Kort, R.; ter Beek, A.

    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

  18. Genotyping of Bacillus anthracis strains based on automated capillary 25-loci Multiple Locus Variable-Number Tandem Repeats Analysis

    Directory of Open Access Journals (Sweden)

    Ciervo Alessandra

    2006-04-01

    Full Text Available Abstract Background The genome of Bacillus anthracis, the etiological agent of anthrax, is highly monomorphic which makes differentiation between strains difficult. A Multiple Locus Variable-number tandem repeats (VNTR Analysis (MLVA assay based on 20 markers was previously described. It has considerable discrimination power, reproducibility, and low cost, especially since the markers proposed can be typed by agarose-gel electrophoresis. However in an emergency situation, faster genotyping and access to representative databases is necessary. Results Genotyping of B. anthracis reference strains and isolates from France and Italy was done using a 25 loci MLVA assay combining 21 previously described loci and 4 new ones. DNA was amplified in 4 multiplex PCR reactions and the length of the resulting 25 amplicons was estimated by automated capillary electrophoresis. The results were reproducible and the data were consistent with other gel based methods once differences in mobility patterns were taken into account. Some alleles previously unresolved by agarose gel electrophoresis could be resolved by capillary electrophoresis, thus further increasing the assay resolution. One particular locus, Bams30, is the result of a recombination between a 27 bp tandem repeat and a 9 bp tandem repeat. The analysis of the array illustrates the evolution process of tandem repeats. Conclusion In a crisis situation of suspected bioterrorism, standardization, speed and accuracy, together with the availability of reference typing data are important issues, as illustrated by the 2001 anthrax letters event. In this report we describe an upgrade of the previously published MLVA method for genotyping of B. anthracis and apply the method to the typing of French and Italian B. anthracis strain collections. The increased number of markers studied compared to reports using only 8 loci greatly improves the discrimination power of the technique. An Italian strain belonging to the

  19. Characterization and Optimization of Amylase Production in WangLB, a High Amylase-Producing Strain of Bacillus.

    Science.gov (United States)

    Wang, Shihui; Jeyaseelan, Jenasia; Liu, Yun; Qin, Wensheng

    2016-09-01

    The costs of amylase represent ca. 24 % of the expenditures in the starch industry and an increase in amylase production and/or activity will greatly cut down on production costs. In the present study, we obtained a high amylase-producing strain of bacteria, WangLB, and identified it as a member of the Bacillus genus based on 16S rDNA analysis. The fermentation conditions for amylase production in the strain were optimized, and the maximum amylase activity we obtained was 26,670 ± 1390 U/mL, under the optimized conditions of 48-h incubation in liquid starch medium, 35 °C, pH 10, 1 % v/v inoculum concentration, 20 g/L starch concentration, and 0.1 % w/v peptone. The influences of 16 small organic inducers on amylase production were tested, and the results showed that 20 mmol/L alanine greatly enhanced amylase production to 290 % of the baseline level. We also conducted an amylase enzymology analysis. The molecular weight of the amylase was 55 kD, determined by SDS-PAGE. The optimum temperature and pH for the amylase were 55 °C and pH 9, respectively. The enzyme also showed high activity over a wide range of temperatures (50-85 °C) and pH values (3-10), and the activity of the amylase was Ca(2+) independent. The kinetic parameters K m and V max were 0.37 ± 0.02 mg/mL and 233 U/mg, respectively. Finally, the amylase was applied to the hydrolysis of five different brands of starch. It was found that the hydrolyzability of the substrate by amylase increased along with starch solubility.

  20. Purification, crystallization, and properties of F1-ATPase complexes from the thermoalkaliphilic Bacillus sp. strain TA2.A1.

    Science.gov (United States)

    Stocker, Achim; Keis, Stefanie; Cook, Gregory M; Dimroth, Peter

    2005-11-01

    Recently, we reported the cloning of the atp operon encoding for the F(1)F(0)-ATP synthase from the extremely thermoalkaliphilic bacterium Bacillus sp. strain TA2.A1. In this study, the genes encoding the F(1) moiety of the enzyme complex were cloned from the atp operon into the vector pTrc99A and expressed in Escherichia coli in two variant complexes, F(1)-wt consisting of subunits alpha(3)beta(3)gammadeltaepsilon and F(1)Deltadelta lacking the entire delta-subunit as a prerequisite for overproduction and crystallization trials. Both F(1)-wt and F(1)Deltadelta were successfully overproduced in E. coli and purified in high yield and purity. F(1)Deltadelta was crystallized by micro-batch screening yielding three-dimensional crystals that diffracted to a resolution of 3.1A using a synchrotron radiation source. After establishing cryo and dehydrating conditions, a complete set of diffraction data was collected from a single crystal. No crystals were obtained with F(1)-wt. Data processing of diffraction patterns showed that F(1)Deltadelta crystals belong to the orthorhombic space group P2(1)2(1)2(1) with unit cell parameters of a=121.70, b=174.80, and c=223.50A, alpha, beta, gamma=90.000. The asymmetric unit contained one molecule of bacterial F(1)Deltadelta with a corresponding volume per protein weight (V(M)) of 3.25A(3) Da(-1) and a solvent content of 62.1%. Silver staining of single crystals of F(1)Deltadelta analyzed by SDS-PAGE revealed four bands alpha, beta, gamma, and epsilon with identical M(r)-values as those found in the native F(1)F(0)-ATP synthase isolated from strain TA2.A1 membranes. ATPase assays of F(1)Deltadelta crystals exhibited latent ATP hydrolytic activity that was highly stimulated by lauryldimethylamine oxide, a hallmark of the native enzyme.

  1. The Correlation of the Presence and Expression Levels of cry Genes with the Insecticidal Activities against Plutella xylostella for Bacillus thuringiensis Strains

    Directory of Open Access Journals (Sweden)

    Ming-Lun Chen

    2014-08-01

    Full Text Available The use of Bacillus thuringiensis (Bt strains with high insecticidal activity is essential for the preparation of bioinsecticide. In this study, for 60 Bt strains isolated in Taiwan, their genotypes and the correlation of some cry genes as well as the expression levels of cry1 genes, with their insecticidal activities against Plutella xylostella, were investigated. Pulsed field gel electrophoresis (PFGE and random amplified polymorphic DNA (RAPD results revealed that the genotypes of these Bt strains are highly diversified. Also, a considerable number of the Bt strains isolated in Taiwan were found to have high insecticidal activities. Since strains that showed individual combined patterns of PFGE and RAPD exhibited distinct insecticidal activities against P. xylostella, thus, these genotypes may be useful for the identification of the new Bt strains and those which have been used in bioinsecticides. In addition, although the presence of cry2Aa1 may have a greater effect on the insecticidal activity of Bt strains in bioassay than other cry genes, only high expression level of cry1 genes plays a key role to determine the insecticidal activity of Bt strains. In conclusion, both RAPD and PFGE are effective in the differentiation of Bt strains. The presence of cry2Aa1 and, especially, the expression level of cry1 genes are useful for the prediction of the insecticidal activities of Bt strains against P. xylostella.

  2. The correlation of the presence and expression levels of cry genes with the insecticidal activities against Plutella xylostella for Bacillus thuringiensis strains.

    Science.gov (United States)

    Chen, Ming-Lun; Chen, Pin-Hsin; Pang, Jen-Chieh; Lin, Chia-Wei; Hwang, Chin-Fa; Tsen, Hau-Yang

    2014-08-19

    The use of Bacillus thuringiensis (Bt) strains with high insecticidal activity is essential for the preparation of bioinsecticide. In this study, for 60 Bt strains isolated in Taiwan, their genotypes and the correlation of some cry genes as well as the expression levels of cry1 genes, with their insecticidal activities against Plutella xylostella, were investigated. Pulsed field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) results revealed that the genotypes of these Bt strains are highly diversified. Also, a considerable number of the Bt strains isolated in Taiwan were found to have high insecticidal activities. Since strains that showed individual combined patterns of PFGE and RAPD exhibited distinct insecticidal activities against P. xylostella, thus, these genotypes may be useful for the identification of the new Bt strains and those which have been used in bioinsecticides. In addition, although the presence of cry2Aa1 may have a greater effect on the insecticidal activity of Bt strains in bioassay than other cry genes, only high expression level of cry1 genes plays a key role to determine the insecticidal activity of Bt strains. In conclusion, both RAPD and PFGE are effective in the differentiation of Bt strains. The presence of cry2Aa1 and, especially, the expression level of cry1 genes are useful for the prediction of the insecticidal activities of Bt strains against P. xylostella.

  3. 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.

  4. Next-Generation Bacillus anthracis Live Attenuated Spore Vaccine Based on the htrA(-) (High Temperature Requirement A) Sterne Strain.

    Science.gov (United States)

    Chitlaru, Theodor; Israeli, Ma'ayan; Bar-Haim, Erez; Elia, Uri; Rotem, Shahar; Ehrlich, Sharon; Cohen, Ofer; Shafferman, Avigdor

    2016-01-06

    Anthrax is a lethal disease caused by the gram-positive spore-producing bacterium Bacillus anthracis. Live attenuated vaccines, such as the nonencapsulated Sterne strain, do not meet the safety standards mandated for human use in the Western world and are approved for veterinary purposes only. Here we demonstrate that disrupting the htrA gene, encoding the chaperone/protease HtrA (High Temperature Requirement A), in the virulent Bacillus anthracis Vollum strain results in significant virulence attenuation in guinea pigs, rabbits and mice, underlying the universality of the attenuated phenotype associated with htrA knockout. Accordingly, htrA disruption was implemented for the development of a Sterne-derived safe live vaccine compatible with human use. The novel B. anthracis SterneΔhtrA strain secretes functional anthrax toxins but is 10-10(4)-fold less virulent than the Sterne vaccine strain depending on animal model (mice, guinea pigs, or rabbits). In spite of this attenuation, double or even single immunization with SterneΔhtrA spores elicits immune responses which target toxaemia and bacteremia resulting in protection from subcutaneous or respiratory lethal challenge with a virulent strain in guinea pigs and rabbits. The efficacy of the immune-protective response in guinea pigs was maintained for at least 50 weeks after a single immunization.

  5. Functional feed assessment on Litopenaeus vannamei using 100% fish meal replacement by soybean meal, high levels of complex carbohydrates and Bacillus probiotic strains.

    Science.gov (United States)

    Olmos, Jorge; Ochoa, Leonel; Paniagua-Michel, Jesus; Contreras, Rosalia

    2011-01-01

    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.

  6. PRODUCTION AND PARTIAL CHARACTERIZATION OF ALKALINE PROTEASE FROM BACILLUS TEQUILENSIS STRAINS CSGAB0139 ISOLATED FROM SPOILT COTTAGE CHEESE

    Directory of Open Access Journals (Sweden)

    Aruna K

    2014-09-01

    Full Text Available An alkaline protease producing strain was isolated from spoilt cottage cheese sample which was identified as Bacillus tequilensis strain SCSGAB0139 on the basis of morphological, cultural, biochemical characteristics and 16S rRNA sequence analysis. Primary screening for protease production was carried out by observing for zone of hydrolysis on skim milk agar, GYEA milk agar and gelatin agar plates. Physicochemical parameters like pH of the medium, incubation time and temperature, aeration and composition of the medium were optimized for maximum protease production by this isolate. Maximum yield of protease (85.67U/ml was obtained in a medium containing peptone (5% w/v, maltose (5% w/v and KNO3, 0.5%; K2HPO4, 0.4%; trisodium citrate, 0.4; CaCl2, 0.0002%; MgSO4·7H2O, 0.05%; Na2CO3, 1%.; 1% (v/v of a trace element solution (NH46MO7O24, 0.01%; FeSO4·7H2O, 0.2%; CuSO4·5H2O, 0.02%; ZnCl2, 0.02% having pH 10, inoculated with 1%(v/v of pre-grown cell mass and incubated at 30°C on a rotary shaker (100rpm for 48hrs. Absence of any one of the following salts viz. KNO3, K2HPO4, tri-sodium citrate; MgSO4, CaCl2 and Na2CO3 from optimized medium reduced the protease production by 80% to 40%. The enzyme has an optimum pH of 9 and maintained its stability over a broad pH range between 6 and 10. Its optimum temperature is 30°C, and exhibited a stability of up to 65°C. Among metal ions only Ca2+ and Mg2+ions enhanced the enzyme activity up to 105% and 107% respectively while other metal ions reduced the activity by 40% where as EDTA exhibited the least inhibitory effect upon the enzyme. Protease activity was enhanced in the presence of isopropanol and marginally reduced in the presence of other organic solvents studied. The crude enzyme showed stability towards various surfactants such as Tween-20, Tween- 80, SDS and Triton X-100. It also showed excellent stability and compatibility with commonly used laundry detergents (Ariel, Surf excel and Surf Blue. The

  7. Comparative transcriptome analysis of the biocontrol strain Bacillus amyloliquefaciens FZB42 as response to biofilm formation analyzed by RNA sequencing.

    Science.gov (United States)

    Kröber, Magdalena; Verwaaijen, Bart; Wibberg, Daniel; Winkler, Anika; Pühler, Alfred; Schlüter, Andreas

    2016-08-10

    The strain Bacillus amyloliquefaciens FZB42 is a plant growth promoting rhizobacterium (PGPR) and biocontrol agent known to keep infections of lettuce (Lactuca sativa) by the phytopathogen Rhizoctonia solani down. Several mechanisms, including the production of secondary metabolites possessing antimicrobial properties and induction of the host plant's systemic resistance (ISR), were proposed to explain the biocontrol effect of the strain. B. amyloliquefaciens FZB42 is able to form plaques (biofilm-like structures) on plant roots and this feature was discussed to be associated with its biocontrol properties. For this reason, formation of B. amyloliquefaciens biofilms was studied at the transcriptional level using high-throughput sequencing of whole transcriptome cDNA libraries from cells grown under biofilm-forming conditions vs. planktonic growth. Comparison of the transcriptional profiles of B. amyloliquefaciens FZB42 under these growth conditions revealed a common set of highly transcribed genes mostly associated with basic cellular functions. The lci gene, encoding an antimicrobial peptide (AMP), was among the most highly transcribed genes of cells under both growth conditions suggesting that AMP production may contribute to biocontrol. In contrast, gene clusters coding for synthesis of secondary metabolites with antimicrobial properties were only moderately transcribed and not induced in biofilm-forming cells. Differential gene expression revealed that 331 genes were significantly up-regulated and 230 genes were down-regulated in the transcriptome of B. amyloliquefaciens FZB42 under biofilm-forming conditions in comparison to planktonic cells. Among the most highly up-regulated genes, the yvqHI operon, coding for products involved in nisin (class I bacteriocin) resistance, was identified. In addition, an operon whose products play a role in fructosamine metabolism was enhanced in its transcription. Moreover, genes involved in the production of the extracellular

  8. Ocorrência de Bacillus cereus em leite integral e capacidade enterotoxigênica das cepas isoladas Occurrence of Bacillus cereus in Whole milk and enterotoxigenic potential of the isolated strains

    Directory of Open Access Journals (Sweden)

    N.C.M. Rezende-Lago

    2007-12-01

    Full Text Available Pesquisaram-se a presença de Bacillus cereus e a produção de enterotoxinas produzidas por esses microrganismos em 120 amostras de diversos tipos de leite. Bacillus cereus foi isolado e identificado em 22 (73,3%, 15 (50,0%, 29 (96,7% e quatro (13,3% amostras de leite em pó, cru, pasteurizado e UAT (longa vida, respectivamente. Para a detecção de enterotoxinas pela técnica da alça ligada de coelho, foram positivos, respectivamente, três (13,6%, um (7,1% e 10 (35,7% microrganismos isolados das amostras de leite em pó, leite cru e leite pasteurizado. Pelo teste de aumento de permeabilidade vascular, dois (9,1%, um (7,1%, um (3,6% e um (4,0% microrganismos isolados de leite em pó, cru, pasteurizado e UAT apresentaram-se enterotoxigênicos, respectivamente. O uso da técnica de aglutinação passiva em látex demonstrou a produção da toxina diarréica por três (33,3%, sete (63,6%, quatro (30,8% e oito (80,0% microrganismos isolados, respectivamente, de leite em pó, cru, pasteurizado e UAT. Os resultados indicam um risco potencial, podendo colocar em risco a saúde dos consumidores desses produtos.A hundred and twenty samples of different types of milk were examined to the presence of Bacillus cereus and the enterotoxigenic potential of the isolated strains. Bacillus cereus was isolated and identified in 22 (73.0%, 15 (50.0%, 29 (96.7% and four (13.3% samples of powder, raw, pasteurized and UHT milk, respectively. The enterotoxigenicity detection using the rabbit ileal loop assay showed positive, respectively, three (13.6%, one (7.1% and 10 (35.7% isolated strains from powder, raw and pasteurized milk. Using vascular permeability activity assay two (9.1%, one (7.1%, one (3.6% and one (4.0% isolated strains from powder, raw, pasteurized and UHT milk were positive, respectively. The reversed passive latex agglutination test showed diarrheal toxin production by three (33.3%, seven (63.6%, four (30.8% and eight (80.0% strains isolated from

  9. Properties of the Bacillus Cereus strain used in probiotic CenBiot Propriedades da cepa de Bacillus cereus utilizada no probiótico CenBiot

    Directory of Open Access Journals (Sweden)

    Carlos Gil-Turnes

    1999-01-01

    Full Text Available Bacillus cereus CenBiot fulfilled the requirements to be used as probiotic. The spores showed D80 of 14 hs, inhibited Escherichia coli and Yersinia pseudotuberculosis after 24 hs in associative culture, were innocuous for suckling and adult mice and were not inhibited by antibiotics at low concentrations.Bacillus cereus CenBiot possui as características necessárias para ser utilizada como probiótico. Os esporos apresentaram D80 de 14 hs, inibiram Escherichia coli e Yersinia pseudotuberculosis após cultivadas associativamente por 24 hs, foram inócuos para camundongos lactentes e adultos e não foram inibidos por antibióticos a baixas concentrações.

  10. In vitro fermentation studies for selection and evaluation of Bacillus strains as starter cultures for the production of okpehe, a traditional African fermented condiment.

    Science.gov (United States)

    Oguntoyinbo, Folarin A; Sanni, Abiodun I; Franz, Charles M A P; Holzapfel, Wilhelm H

    2007-01-25

    Selected Bacillus and Enterococcus strains, isolated from traditional okpehe fermentations, were studied for their suitability as starter cultures in laboratory-scale fermentations of Prosopis africana seeds for the production of okpehe, a traditional fermented vegetable product of Nigeria. The strains were selected on the basis of highest proteolytic activity, as determined with the APIZYM (BioMerieux) test. The choice of starter strains was narrowed to Bacillus subtilis strains BFE 5301 and BFE 5372. These were determined as the best starter combination because of rapid growth, high amylolytic and proteolytic activities, high levels of polyglutamic acid production by strain BFE 5372, as well as bacteriocin production by strain BFE 5301. Other mixed culture fermentations did not yield sensorically acceptable products. Although a monoculture fermentation, using only B. subtilis strain BFE 5372, produced okpehe with very good sensory characteristics, the growth of B. cereus could be detected after 48 h fermentation, indicating that this starter did not sufficiently contribute to product safety. Mixed culture fermentation with the combination of bacteriocin-producing starter B. subtilis BFE 5301 and the non-bacteriocin-producing B. subtilis BFE 5372, produced a product with good sensory characteristics, in which growth of B. cereus was delayed. The bacteriocin produced by B. subtilis strain BFE 5301 was identified as subtilisin, using subtilisin-specific primers and PCR amplification of the subtilisin gene. The bacteriocin was heat-stable at 100 degrees C for 10 min and exhibited highest activity at pH values lower or equal to pH 6.0. The bacteriocin was sensitive to the proteolytic enzymes trypsin and alpha-chymotrypsin at concentrations of 10 mg/ml.

  11. Influence of nitrogen source and pH value on undesired poly(γ-glutamic acid) formation of a protease producing Bacillus licheniformis strain.

    Science.gov (United States)

    Meissner, Lena; Kauffmann, Kira; Wengeler, Timo; Mitsunaga, Hitoshi; Fukusaki, Eiichiro; Büchs, Jochen

    2015-09-01

    Bacillus spp. are used for the production of industrial enzymes but are also known to be capable of producing biopolymers such as poly(γ-glutamic acid). Biopolymers increase the viscosity of the fermentation broth, thereby impairing mixing, gas/liquid mass and heat transfer in any bioreactor system. Undesired biopolymer formation has a significant impact on the fermentation and downstream processing performance. This study shows how undesirable poly(γ-glutamic acid) formation of an industrial protease producing Bacillus licheniformis strain was prevented by switching the nitrogen source from ammonium to nitrate. The viscosity was reduced from 32 to 2.5 mPa s. A constant or changing pH value did not influence the poly(γ-glutamic acid) production. Protease production was not affected: protease activities of 38 and 46 U mL(-1) were obtained for ammonium and nitrate, respectively. With the presented results, protease production with industrial Bacillus strains is now possible without the negative impact on fermentation and downstream processing by undesired poly(γ-glutamic acid) formation.

  12. Isolation, identification and antimicrobial activity screening of endophytic bacteria Bacillus megaterium z12 strain from bryophytes%苔藓内生菌Bacillus megaterium z12的分离、鉴定和抗菌活性筛选

    Institute of Scientific and Technical Information of China (English)

    张鑫; 王婷; 崔灵芝; 刘新利

    2012-01-01

    Objective An endophytic bacteria zl2 was isolated from the leaf of Plagiochasma intermedium which has broad-spectrum antimicrobial activity. Methods According to the mophology, physiological and biochemical characteristics, combined with phylogenetic analysis, and the antimicrobial activity of zl2 was analyzed with 14 tested strains. Results The bacteria was identified as Bacillus megaterium. The results of inhibition experiments showed that zl2 strain had broad-spectrum antimicrobial activity to 7 target strains in different degrees, especially for Staphylococcus aureus and Candida albicans, Corticium sasakii were investigated by further tests with crude extracts of zl2 strain fermentation. Conclusion The strain is deserved to develop an agricultural antibiotics.%目的 从无纹紫背苔(Plagiochasma intermedium)叶片中分离筛选出一株具广谱抗菌活性的内生菌z12.方法 根据菌落和细胞形态、生理生化特性,以及16S rDNA序列对内生细菌z12进行了分析鉴定,并研究了它对14种供试菌株的抑制作用.结果 经分析鉴定该菌属于巨大芽孢杆菌(Bacillus megaterium).抑菌实验表明,内生细菌z12对7种供试菌都有不同程度的抑制作用,对金黄色葡萄球菌(Staphylococcus aureus)和白念珠菌(Candida albicans)的抑制作用较强.研究了z12菌株粗提物对水稻纹枯病原菌(Corticium sasakii)的抑制作用.结论 该菌株具有开发成农用抗生素的价值.

  13. THE KINETICS OF THE REACTIONS CATALYZED BY AN ENZYMATIC PREPARATION PRODUCED BY A BACILLUS LICHENIFORMIS STRAIN

    Directory of Open Access Journals (Sweden)

    DRAGOMIRESCU MONICA

    2007-01-01

    Full Text Available Robust immobilization techniques that preserve the activity of biomolecules havemany potential applications. In recent years, a number of new bioimobilisationmethods in sol-gel-derived materials were reported. The interactions between thebiomolecule and the inorganic material determine the degree to which thebiomolecule retains its native properties. The newer technological developments inthe field of immobilized biocatalysts can offer the possibility of a wider and moreeconomical exploitation of biocatalysts in biological applications, food and feedindustry, medicine, and in the development of bioprocess monitoring devices, like thebiosensors.The aim of this study was to obtain immobilized enzymatic preparations by methodswhich affect enzyme conformations and kinetic parameters as less as possible. Weimmobilized the enzymatic preparation with protease activity produced by a Bacilluslicheniformis B 40 local strain by physical bonding on ceramics and entrapment intosol-gel-derived glasses obtained from tetraethyl orthosilicate (TEOS, deposited inthin layer on a ceramic support (entrapment/deposition. Both physically adsorbedand entrapped/deposited enzymes follow Michaelis-Menten kinetics, similar with thesoluble enzyme. In the case of immobilized enzymes, the apparent Michaelisconstant, Km, was greater than that of the native one, as it was expected. The kineticparameters indicate that the enzymatic preparations adsorbed on ceramic supportand entrapped/deposited show less affinity for the substrate, Km being 1.3 and 2.1times higher than that of the native enzyme, respectively. The maximum velocityincreased also by 3.5 and 7.9 times respectively, compared with the free counterpart(according to Lineweaver-Burk linearization.

  14. Comparative Study on Biological Characteristics of Six Bacillus Strains%6株芽孢杆菌的生物学特性比较研究

    Institute of Scientific and Technical Information of China (English)

    李雅丽; 秦艳; 周绪霞; 李卫芬

    2011-01-01

    The main biological characteristics of six Bacillus strains isolated were studied,including the activity of enzyme-producing,the endurance to NaCl,bile salt and artificial gastric juice as well as artificial intestinal fluid and the sensitivity to antibiotics. The results showed that,Bacillus subtilis strain 1, B. coagulans and B. cereus had the capability of producing high yield of protease and amylase. Two strains of B. subtilis and B. coagulans showed well resistance to high concentration of NaCl.B. subtilis strain 2,B. coagulans and B. natto as well as B. cereus had relatively strong tolerance for bovine bile salt. Except B. licheniformis,the other five strains of Bacillus could resist the artificial gastric and intestinal fluid well. Six strains of Bacillus were sensitive to common forage antibiotics. These results showedthat B. subtilis 1, B. coagulans and B. cereus were excellent probiotics.%以实验室分离保存的6株芽孢杆菌为材料,对其主要生物学特性进行了研究,包括产酶活力,对NaCl、牛胆盐、人工胃液和人工肠液的耐受性及对抗生素的敏感性.结果显示.枯草芽孢杆菌1、凝结芽孢杆菌和蜡样芽孢杆菌均能产生较高活性的中性蛋白酶和淀粉酶;2株枯草芽孢杆菌和凝结芽孢杆菌能耐较高浓度的NaCl;枯草芽孢杆菌2、凝结、纳豆和蜡样芽孢杆菌对牛胆盐的耐受性较强;除地衣芽孢杆菌外,其他5株芽孢杆菌对胃液和肠液的耐受性较好;6株芽孢杆菌均对常用饲用抗生素敏感.结果表明,枯草1、凝结、蜡样芽孢杆菌是优良的候选益生菌菌株.

  15. The mechanism of uranium transformation from U(VI) into nano-uramphite by two indigenous Bacillus thuringiensis strains

    Energy Technology Data Exchange (ETDEWEB)

    Pan, Xiaohong; Chen, Zhi [Key Lab of Biopesticide and Chemical Biology, Fujian Agriculture and Forestry University, Ministry of Education & Fujian–Taiwan Joint Center for Ecological Control of Crop Pests, Fuzhou, Fujian 350002 (China); Key Laboratory of Design and Assembly of Functional Nanostructures, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian 350002 (China); Chen, Fanbing [Key Lab of Biopesticide and Chemical Biology, Fujian Agriculture and Forestry University, Ministry of Education & Fujian–Taiwan Joint Center for Ecological Control of Crop Pests, Fuzhou, Fujian 350002 (China); Cheng, Yangjian [Key Laboratory of Design and Assembly of Functional Nanostructures, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian 350002 (China); Lin, Zhang, E-mail: zlin@fjirsm.ac.cn [Key Laboratory of Design and Assembly of Functional Nanostructures, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian 350002 (China); School of Environment and Energy, South China University of Technology, Guangzhou 510006 (China); Guan, Xiong, E-mail: guanxfafu@126.com [Key Lab of Biopesticide and Chemical Biology, Fujian Agriculture and Forestry University, Ministry of Education & Fujian–Taiwan Joint Center for Ecological Control of Crop Pests, Fuzhou, Fujian 350002 (China)

    2015-10-30

    Highlights: • Indigenous B. thuringiensis exhibited highly accumulation ability to U(VI) in the absence of additional nutrients. • The amorphous uranium compound would transformed into crystalline nano-uramphite by B. thuringiensis. • The chemical nature of formed U-species were monitored. • The cell-free extracts of B. thuringiensis had better uranium-immobilization ability than its cell debris. • Provided the understanding of the uranium transformation mechanism. - Abstract: The mechanism of uranium transformation from U(VI) into nano-uramphite by two indigenous Bacillus thuringiensis strains was investigated in the present work. Our data showed that the bacteria isolated from uranium mine possessed highly accumulation ability to U(VI), and the maximum accumulation capacity was around 400 mg U/g biomass (dry weight). X-ray powder diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FT-IR) analyzes indicated that the U(VI) was adsorbed on the bacterial surface firstly through coordinating with phosphate, −CH{sub 2} and amide groups, and then needle-like amorphous uranium compounds were formed. With the extension of time, the extracellular crystalline substances were disappeared, but some particles were appeared in the intracellular region, and these particles were characterized as tetragonal-uramphite. Moreover, the disrupted experiment indicated that the cell-free extracts had better uranium-immobilization ability than cell debris. Our findings provided the understanding of the uranium transformation process from amorphous uranium to crystalline uramphite, which would be useful in the regulation of uranium immobilization process.

  16. Characteristics of a Novel Highly Thermostable and Extremely Thermophilic Alkalitolerant Amylase from Hyperthermophilic Bacillus Strain HUTBS71

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    Hazem Akel

    2009-01-01

    Full Text Available Problem statement: This study reported the purification and characterization of a novel highly thermostable alkaline amylase from a newly isolated Bacillus strain HUTBS71. Approach: The enzyme was purified using ammonium sulfate precipitation, ion exchange and gel filtration chromatography. Results: Maximum amylase activity (72 U mL-1 was obtained at 100°C after 10 min of incubation. The enzyme was purified 24 fold with 12.5% yield and showed a monomer band with a molecular weight of 58.8 kDa by SDS-PAGE. This enzyme exhibited maximum activity at pH and temperature, 7.8 and 100°C, respectively. It performed stability over a broad range of pH and temperature, 5.2-10.0 and 80-115°C, respectively. The half-life of the enzyme at 90 and 100°C was estimated to be 3 h. The activation energy of denaturation of purified enzyme was 2.53 kJ moL-1. The enzyme was activated by 5 mM of CoCl2, MgSO4, MnCl2, ZnSO4 and MnSO4 (relative activity was 133, 126, 133, 106.6 and 103%, respectively. It was strongly inhibited by CuSO4 and CdCl2 but less affected by NaCl, CaCl2, FeCl3, ZnCl2 and EDTA. Conclusion: The present purified amylase therefore could be defined as a highly thermostable, extremely hyperthermophilic and alkalitolerant with new properties make the present enzyme applicable for many starch processing and food industries.

  17. Bacillus velezensis is not a later heterotypic synonym of Bacillus amyloliquefaciens; Bacillus methylotrophicus, Bacillus amyloliquefaciens subsp plantarum and ‘Bacillus oryzicola’ are later heterotypic synonyms of Bacillus

    Science.gov (United States)

    The rhizosphere isolated bacteria belonging to the Bacillus amyloliquefaciens subsp. plantarum and Bacillus methylotrophicus clades are an important group of strains that are used as plant growth promoters and antagonists of plant pathogens. These properties have made these strains the focus of comm...

  18. Characterization of three Bacillus cereus strains involved in a major outbreak of food poisoning after consumption of fermented black beans (Douchi) in Yunan, China.

    Science.gov (United States)

    Zhou, Guoping; Bester, Kai; Liao, Bin; Yang, Zushun; Jiang, Rongrong; Hendriksen, Niels Bohse

    2014-10-01

    Three Bacillus cereus strains isolated from an outbreak of food poisoning caused by the consumption of fermented black beans (douchi) containing B. cereus is described. The outbreak involved 139 persons who had nausea, vomiting, and diarrhea. The strains were isolated from vomit and the unprepared douchi. Two of the strains produced the emetic toxin cereulide, as evidenced by polymerase chain reaction analysis for the presence of the nonribosomal synthetase cluster responsible for the synthesis of cereulide and by chemical analysis by high-performance liquid chromatography-mass spectrometry. These two strains belong to genetic group III of B. cereus, and multiple locus sequence typing revealed that the type was ST26, as a major part of B. cereus emetic strains. One of these strains produced significantly more cereulide at 37°C than the type cereulide producer (F4810/72), and it was also able to produce the toxin at 40°C and 42°C. The third strain belongs to genetic group IV, and it is a new multiple locus sequence type closely related to strains that are cytotoxic and enterotoxigenic. It possesses genes for hemolysin BL, nonhemolytic enterotoxin, and cytotoxin K2; however, it varies from the majority of strains possessing genes for hemolysin BL by not being hemolytic. Thus, two B. cereus strains producing the emetic toxin cereulide and a strain producing enterotoxins might have been involved in this food-poisoning incident caused by the consumption of a natural fermented food. The ability of one of the strains to produce cereulide at ≥37°C makes it possible that it is produced in the human gut in addition to occurring in the food.

  19. Structural elucidation of the nonclassical secondary cell wall polysaccharide from Bacillus cereus ATCC 10987. Comparison with the polysaccharides from Bacillus anthracis and B. cereus type strain ATCC 14579 reveals both unique and common structural features.

    Science.gov (United States)

    Leoff, Christine; Choudhury, Biswa; Saile, Elke; Quinn, Conrad P; Carlson, Russell W; Kannenberg, Elmar L

    2008-10-31

    Nonclassical secondary cell wall polysaccharides constitute a major cell wall structure in the Bacillus cereus group of bacteria. The structure of the secondary cell wall polysaccharide from Bacillus cereus ATCC 10987, a strain that is closely related to Bacillus anthracis, was determined. This polysaccharide was released from the cell wall with aqueous hydrogen fluoride (HF) and purified by gel filtration chromatography. The purified polysaccharide, HF-PS, was characterized by glycosyl composition and linkage analyses, mass spectrometry, and one- and two-dimensional NMR analysis. The results showed that the B. cereus ATCC 10987 HF-PS has a repeating oligosaccharide consisting of a -->6)-alpha-GalNAc-(1-->4)-beta-ManNAc-(1-->4)-beta-GlcNAc-(1--> trisaccharide that is substituted with beta-Gal at O3 of the alpha-GalNAc residue and nonstoichiometrically acetylated at O3 of the N-acetylmannosamine (ManNAc) residue. Comparison of this structure with that of the B. anthracis HF-PS and with structural data obtained for the HF-PS from B. cereus type strain ATCC 14579 revealed that each HF-PS had the same general structural theme consisting of three HexNAc and one Hex residues. A common structural feature in the HF-PSs from B. cereus ATCC 10987 and B. anthracis was the presence of a repeating unit consisting of a HexNAc(3) trisaccharide backbone in which two of the three HexNAc residues are GlcNAc and ManNAc and the third can be either GlcNAc or GalNAc. The implications of these results with regard to the possible functions of the HF-PSs are discussed.

  20. Proteome analysis of a recombinant Bacillus megaterium strain during heterologous production of a glucosyltransferase

    Directory of Open Access Journals (Sweden)

    Jahn Dieter

    2005-05-01

    Full Text Available Abstract A recombinant B. megaterium strain was used for the heterologous production of a glucosyltransferase (dextransucrase. To better understand the physiological and metabolic responses of the host cell to cultivation and induction conditions, proteomic analysis was carried out by combined use of two-dimensional gel electrophoresis and mass spectrometry (2-DE/MS for protein separation and identification. 2-DE method was optimized for the separation of intracellular proteins. Since the genome of B. megaterium is not yet available, peptide sequencing using peptide fragment information obtained from nanoelectrospray ionization quadrupole-time-of-flight tandem mass spectrometry (ESI-QqTOF MS/MS was applied for protein identification. 167 protein spots were identified as 149 individual proteins, including most enzymes involved in the central carbon metabolic pathways and many enzymes related to amino acid synthesis and protein synthesis. Based on the results a 2-DE reference map and a corresponding protein database were constructed for further proteomic approaches on B. megaterium. For the first time it became possible to perform comparative proteomic analysis on B. megaterium in a batch culture grown on glucose with xylose induction for dextrasucrase production. No significant differences were observed in the expression changes of enzymes of the glycolysis and TCA cycle, indicating that dextransucrase production, which amounted to only 2 % of the entire protein production, did not impose notable metabolic or energetic burdens on the central carbon metabolic pathway of the cells. However, a short-term up-regulation of aspartate aminotransferase, an enzyme closely related to dextransucrase production, in the induced culture demonstrated the feasibility to use 2-DE method for monitoring dextransucrase production. It was also observed that under the cultivation conditions used in this study B. megaterium tended to channel acetyl-CoA into pathways of

  1. Disruption of microbial biofilms by an extracellular protein isolated from epibiotic tropical marine strain of Bacillus licheniformis

    Digital Repository Service at National Institute of Oceanography (India)

    Dusane, D.H.; Damare, S.R.; Nancharaiah, Y.V.; Ramaiah, N.; Venugopalan, V.P.; Kumar, A.R.; Zinjarde, S.S.

    from the surface of green mussel, Perna viridis showed antimicrobial activity against pathogenic Candida albicans BH, Pseudomonas aeruginosa PAO1 and biofouling Bacillus pumilus TiO1 cultures. The antimicrobial activity was lost after treatment...

  2. Different Transcriptional Profiles of Human Monocyte-Derived Dendritic Cells Infected with Distinct Strains of Mycobacterium tuberculosis and Mycobacterium bovis Bacillus Calmette-Guérin

    Directory of Open Access Journals (Sweden)

    Nunzia Sanarico

    2011-01-01

    Full Text Available In order to analyze dendritic cells (DCs activation following infection with different mycobacterial strains, we studied the expression profiles of 165 genes of human monocyte-derived DCs infected with H37Rv, a virulent Mycobacterium tuberculosis (MTB laboratory strain, CMT97, a clinical MTB isolate, Mycobacterium bovis bacillus Calmette-Guérin (BCG, Aventis Pasteur, and BCG Japan, both employed as vaccine against tuberculosis. The analysis of the gene expression reveals that, despite a set of genes similarly modulated, DCs response resulted strain dependent. In particular, H37Rv significantly upregulated EBI3 expression compared with BCG Japan, while it was the only strain that failed to release a significant IL-10 amount. Of note, BCG Japan showed a marked increase in CCR7 and TNF-α expression regarding both MTB strains and it resulted the only strain failing in exponential intracellular growth. Our results suggest that DCs display the ability to elicit a tailored strain-specific immune response.

  3. Significance of Bacillus subtilis strain SJ-101 as a bioinoculant for concurrent plant growth promotion and nickel accumulation in Brassica juncea.

    Science.gov (United States)

    Zaidi, Sabina; Usmani, Saima; Singh, Braj Raj; Musarrat, Javed

    2006-08-01

    In this study, a nickel (Ni)-tolerant Bacillus subtilis strain SJ-101 was characterized based on the 16SrDNA homology and phylogenetic analysis. The role of this strain ascertained in facilitating Ni accumulation in the Indian mustard plant (Brassica juncea [L]. Czern and Coss) var. Pusa Bold (DIR-50), to elucidate the potential of Ni phytoremediation in combination with metal-tolerant rhizobacteria. The data revealed that the plants exposed to NiCl2 (1750 mg kg(-1)) in soil bioaugmented with strain SJ-101 have accumulated 0.147% Ni vis-à-vis 0.094% accumulation in dry biomass of the plants grown in uninoculated soil. The strain SJ-101 has also exhibited the capability of producing indole acetic acid (IAA) (55 microg ml(-1)), and solubilizing inorganic phosphate (90 microg ml(-1)) in specific culture media. The pot culture experiments clearly demonstrated the beneficial effects of bioinoculant strain SJ-101 with significant increase (pplant growth attributes in untreated control soil. Furthermore, the protective effect of the strain SJ-101 against Ni phytotoxicity was evident in plants grown in soil treated with NiCl2 in concentration range of 250-1750 mg kg(-1). Thus, it is suggested that the strain SJ-101 owing to its intrinsic abilities of plant growth promotion, and attenuation of soil Ni by biosorption and bioaccumulation, could be exploited for bacteria-assisted phytoaccumulation of this toxic heavy metal from contaminated sites.

  4. Genomic analysis of three African strains of Bacillus anthracis demonstrates that they are part of the clonal expansion of an exclusively pathogenic bacterium

    Directory of Open Access Journals (Sweden)

    L. Rouli

    2014-11-01

    Full Text Available Bacillus anthracis is the causative agent of anthrax and is classified as a ‘Category A’ biological weapon. Six complete genomes of B. anthracis (A0248, Ames, Ames Ancestor, CDC684, H0491, and Sterne are currently available. In this report, we add three African strain genomes: Sen2Col2, Sen3 and Gmb1. To study the pan‐genome of B. anthracis, we used bioinformatics tools, such as Cluster of Orthologous Groups, and performed phylogenetic analysis. We found that the three African strains contained the pX01 and pX02 plasmids, the nonsense mutation in the plcR gene and the four known prophages. These strains are most similar to the CDC684 strain and belong to the A cluster. We estimated that the B. anthracis pan‐genome has 2893 core genes (99% of the genome size and 85 accessory genes. We validated the hypothesis that B. anthracis has a closed pan‐genome and found that the three African strains carry the two plasmids associated with bacterial virulence. The pan‐genome nature of B. anthracis confirms its lack of exchange (similar to Clostridium tetani and supports its exclusively pathogenic role, despite its survival in the environment. Moreover, thanks to the study of the core content single nucleotide polymorphisms, we can see that our three African strains diverged very recently from the other B. anthracis strains.

  5. [Antagonistic activity of Bacillus amyloliquefaciens subsp. plantarum IMV B-7404 and BIM B-439D strains towards pathogenic bacteria and micromycetes].

    Science.gov (United States)

    Avdeeva, L V; Dragovoz, I V; Korzh, Iu V; Leonova, N O; Iutinskaia, G A; Berezhnaia, A V; Kuptsov, V N; Mandrik, M N; Kolomiets, É I

    2014-01-01

    In this study the antagonistic activity of strains Bacillus amyloliquefaciens subsp. plantarum IMV B-7404 and BIM B-439D against bacterial and fungal pathogens of agricultural crops has been researched. It is shown that both strains of bacilli demonstrated a high level of antagonism to the vascular bacteriosis pathogen, average level of antagonism to micromycetes--root rot pathogens. To ofiobulez pathogen strain B. amyloliquefaciens subsp. plantarum BIM B-439D was more active. Cultural liquid of this strain effectively inhibited the spore's germination of pathogenic micromycetes Penicillium expansum and Botrytis cinerea. Both strains of bacilli synthesized several hydrolytic exoenzymes: proteases, amylases, β-glucanases, chitinases and xylanases. The obtained data suggest the possibility of expanding the range of strain B. amyloliquefaciens subsp. plantarum BIM B-439D application for plant protection, as well as the need for further researches of the exometabolites spectrum of strain B. amyloliquefaciens subsp. plantarum IMV B-7404 and their biological activity in order to create an effective bioformulation for crop protection.

  6. Bacillus sp. strain P38: an efficient producer of L-lactate from cellulosic hydrolysate, with high tolerance for 2-furfural.

    Science.gov (United States)

    Peng, Lili; Wang, Limin; Che, Chengchuan; Yang, Ge; Yu, Bo; Ma, Yanhe

    2013-12-01

    In this study, efficient polymer-grade L-lactic acid production was achieved with the strain Bacillus sp. P38 by using cellulosic hydrolysate as the sole carbon source. In fed-batch fermentation, 180 g L(-1)L-lactic acid was obtained with a volumetric productivity of 2.4 g L(-1)h(-1) and a yield of 0.96 g g(-1) total reducing sugars. No D-isomer of lactic acid was detected in the broth. Strain P38 tolerated up to 10 g L(-1) 2-furfural, and lactate production was sharply inhibited only when the 2-furfural concentration was higher than 6 g L(-1). Moreover, strain P38 also tolerated high concentrations (>6 g L(-1)) of other fermentation inhibitors in cellulosic hydrolysate, such as vanillin and acetic acid, although it was slightly sensitive to formic acid. The efficient L-lactic acid production, combined with high inhibitor tolerance and efficient pentose utilization, indicate that Bacillus sp. P38 is a promising producer of polymer-grade L-lactic acid from cellulosic biomass.

  7. Assessment of Bacillus thuringiensis subsp. israelensis Strain AM65-52 aqueous suspension for the control of black fly populations, (Simulium spp.) in northern Italy

    OpenAIRE

    P. Radeghieri; Santi, F.; Maini, S.

    2013-01-01

    Four field trials were conducted in Mantua province in northern Italy to evaluate the effect of the Bacillus thuringiensis subsp. israelensis Strain AM65-52 based VectoBac® 12 AS larvicide on black flies. Our objective was to examine and compare the efficacy of different concentrations of this microbial larvicide in small plain streams. Larvicidal activity was exhibited in all trials. VectoBac® 12 AS at 2.5 ppm/min rate generated larval mortality from the treatment point to 500 meters downstr...

  8. Comparative genome analysis of Bacillus cereus group genomes with Bacillus subtilis

    OpenAIRE

    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

    2005-01-01

    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-...

  9. Plant growth-promoting rhizobacteria strain Bacillus amyloliquefaciens NJN-6-enriched bio-organic fertilizer suppressed Fusarium wilt and promoted the growth of banana plants.

    Science.gov (United States)

    Yuan, Jun; Ruan, Yunze; Wang, Beibei; Zhang, Jian; Waseem, Raza; Huang, Qiwei; Shen, Qirong

    2013-04-24

    Bacillus amyloliquefaciens strain NJN-6 is an important plant growth-promoting rhizobacteria (PGPR) which can produce secondary metabolites antagonistic to several soil-borne pathogens. In this study, the ability of a bio-organic fertilizer (BIO) containing NJN-6 strain to promote the growth and suppress Fusarium wilt of banana plants was evaluated in a pot experiment. The results showed that the application of BIO significantly decreased the incidence of Fusarium wilt and promoted the growth of banana plants compared to that for the organic fertilizer (OF). To determine the beneficial mechanism of the strain, the colonization of NJN-6 strain on banana roots was evaluated using scanning electron microscopy (SEM). The plant growth-promoting hormones indole-3-acetic acid (IAA) and gibberellin A3 (GA3), along with antifungal lipopeptides iturin A, were detected when the NJN-6 strain was incubated in both Landy medium with additional l-tryptophan and in root exudates of banana plants. In addition, some antifungal volatile organic compounds and iturin A were also detected in BIO. In summary, strain NJN-6 could colonize the roots of banana plants after the application of BIO and produced active compounds which were beneficial for the growth of banana plants.

  10. Isolation and characterization of a new Bacillus thuringiensis strain Lip harboring a new cry1Aa gene highly toxic to Ephestia kuehniella (Lepidoptera: Pyralidae) larvae.

    Science.gov (United States)

    El Khoury, Micheline; Azzouz, Hichem; Chavanieu, Alain; Abdelmalak, Nouha; Chopineau, Joël; Awad, Mireille Kallassy

    2014-06-01

    The aim of this study was to characterize new Bacillus thuringiensis strains that have a potent insecticidal activity against Ephestia kuehniella larvae. Strains harboring cry1A genes were tested for their toxicity, and the Lip strain showed a higher insecticidal activity compared to that of the reference strain HD1 (LC50 of Lip and HD1 were 33.27 and 128.61 μg toxin/g semolina, respectively). B. thuringiensis Lip harbors and expresses cry1Aa, cry1Ab, cry1Ac, cry1Ad and cry2A. DNA sequencing revealed several polymorphisms in Lip Cry1Aa and Cry1Ac compared to the corresponding proteins of HD1. The activation process using Ephestia kuehniella midgut juice showed that Lip Cry1A proteins were more stable in the presence of larval proteases. Moreover, LipCry1A proteins exhibited higher insecticidal activity against these larvae. These results indicate that Lip is an interesting strain that could be used as an alternative to the worldwide used strain HD1.

  11. Biocontrol traits and antagonistic potential of Bacillus amyloliquefaciens strain NJZJSB3 against Sclerotinia sclerotiorum, a causal agent of canola stem rot.

    Science.gov (United States)

    Wu, Yuncheng; Yuan, Jun; Raza, Waseem; Shen, Qirong; Huang, Qiwei

    2014-10-01

    Bacillus amyloliquefaciens strain NJZJSB3 has shown antagonism of several phytopathogens in vitro, especially Sclerotinia sclerotiorum. Both the broth culture and cell suspension of strain NJZJSB3 could completely protect the detached leaves of canola (Brassica napus) from S. sclerotiorum infection. In pot experiments, the application of strain NJZJSB3 cell suspension (10(8) CFU/ml) decreased the disease incidence by 83.3%, a result similar to commercially available fungicide (Dimetachlone). In order to investigate the potential biocontrol mechanisms of strain NJZJSB3, the nonvolatile antifungal compounds it produces were identified as iturin homologs using HPLC-ESI-MS. Antifungal volatile organic compounds were identified by gas chromatography-mass spectrometry. The detected volatiles toluene, phenol, and benzothiazole showed antifungal effects against S. sclerotiorum in chemical control experiments. Strain NJZJSB3 also produced biofilm, siderophores and cell-wall-degrading enzymes (protease and β-1,3-glucanase). These results suggest that strain NJZJSB3 can be a tremendous potential agent for the biological control of sclerotinia stem rot.

  12. Draft genome sequences of two Bacillus thuringiensis strains and characterization of a putative 41.9-kDa insecticidal toxin.

    Science.gov (United States)

    Palma, Leopoldo; Muñoz, Delia; Berry, Colin; Murillo, Jesús; Caballero, Primitivo

    2014-04-30

    In this work, we report the genome sequencing of two Bacillus thuringiensis strains using Illumina next-generation sequencing technology (NGS). Strain Hu4-2, toxic to many lepidopteran pest species and to some mosquitoes, encoded genes for two insecticidal crystal (Cry) proteins, cry1Ia and cry9Ea, and a vegetative insecticidal protein (Vip) gene, vip3Ca2. Strain Leapi01 contained genes coding for seven Cry proteins (cry1Aa, cry1Ca, cry1Da, cry2Ab, cry9Ea and two cry1Ia gene variants) and a vip3 gene (vip3Aa10). A putative novel insecticidal protein gene 1143 bp long was found in both strains, whose sequences exhibited 100% nucleotide identity. The predicted protein showed 57 and 100% pairwise identity to protein sequence 72 from a patented Bt strain (US8318900) and to a putative 41.9-kDa insecticidal toxin from Bacillus cereus, respectively. The 41.9-kDa protein, containing a C-terminal 6× HisTag fusion, was expressed in Escherichia coli and tested for the first time against four lepidopteran species (Mamestra brassicae, Ostrinia nubilalis, Spodoptera frugiperda and S. littoralis) and the green-peach aphid Myzus persicae at doses as high as 4.8 µg/cm2 and 1.5 mg/mL, respectively. At these protein concentrations, the recombinant 41.9-kDa protein caused no mortality or symptoms of impaired growth against any of the insects tested, suggesting that these species are outside the protein's target range or that the protein may not, in fact, be toxic. While the use of the polymerase chain reaction has allowed a significant increase in the number of Bt insecticidal genes characterized to date, novel NGS technologies promise a much faster, cheaper and efficient screening of Bt pesticidal proteins.

  13. Draft Genome Sequences of Two Bacillus thuringiensis Strains and Characterization of a Putative 41.9-kDa Insecticidal Toxin

    Science.gov (United States)

    Palma, Leopoldo; Muñoz, Delia; Berry, Colin; Murillo, Jesús; Caballero, Primitivo

    2014-01-01

    In this work, we report the genome sequencing of two Bacillus thuringiensis strains using Illumina next-generation sequencing technology (NGS). Strain Hu4-2, toxic to many lepidopteran pest species and to some mosquitoes, encoded genes for two insecticidal crystal (Cry) proteins, cry1Ia and cry9Ea, and a vegetative insecticidal protein (Vip) gene, vip3Ca2. Strain Leapi01 contained genes coding for seven Cry proteins (cry1Aa, cry1Ca, cry1Da, cry2Ab, cry9Ea and two cry1Ia gene variants) and a vip3 gene (vip3Aa10). A putative novel insecticidal protein gene 1143 bp long was found in both strains, whose sequences exhibited 100% nucleotide identity. The predicted protein showed 57 and 100% pairwise identity to protein sequence 72 from a patented Bt strain (US8318900) and to a putative 41.9-kDa insecticidal toxin from Bacillus cereus, respectively. The 41.9-kDa protein, containing a C-terminal 6× HisTag fusion, was expressed in Escherichia coli and tested for the first time against four lepidopteran species (Mamestra brassicae, Ostrinia nubilalis, Spodoptera frugiperda and S. littoralis) and the green-peach aphid Myzus persicae at doses as high as 4.8 µg/cm2 and 1.5 mg/mL, respectively. At these protein concentrations, the recombinant 41.9-kDa protein caused no mortality or symptoms of impaired growth against any of the insects tested, suggesting that these species are outside the protein’s target range or that the protein may not, in fact, be toxic. While the use of the polymerase chain reaction has allowed a significant increase in the number of Bt insecticidal genes characterized to date, novel NGS technologies promise a much faster, cheaper and efficient screening of Bt pesticidal proteins. PMID:24784323

  14. Draft Genome Sequences of Two Bacillus thuringiensis Strains and Characterization of a Putative 41.9-kDa Insecticidal Toxin

    Directory of Open Access Journals (Sweden)

    Leopoldo Palma

    2014-04-01

    Full Text Available In this work, we report the genome sequencing of two Bacillus thuringiensis strains using Illumina next-generation sequencing technology (NGS. Strain Hu4-2, toxic to many lepidopteran pest species and to some mosquitoes, encoded genes for two insecticidal crystal (Cry proteins, cry1Ia and cry9Ea, and a vegetative insecticidal protein (Vip gene, vip3Ca2. Strain Leapi01 contained genes coding for seven Cry proteins (cry1Aa, cry1Ca, cry1Da, cry2Ab, cry9Ea and two cry1Ia gene variants and a vip3 gene (vip3Aa10. A putative novel insecticidal protein gene 1143 bp long was found in both strains, whose sequences exhibited 100% nucleotide identity. The predicted protein showed 57 and 100% pairwise identity to protein sequence 72 from a patented Bt strain (US8318900 and to a putative 41.9-kDa insecticidal toxin from Bacillus cereus, respectively. The 41.9-kDa protein, containing a C-terminal 6× HisTag fusion, was expressed in Escherichia coli and tested for the first time against four lepidopteran species (Mamestra brassicae, Ostrinia nubilalis, Spodoptera frugiperda and S. littoralis and the green-peach aphid Myzus persicae at doses as high as 4.8 µg/cm2 and 1.5 mg/mL, respectively. At these protein concentrations, the recombinant 41.9-kDa protein caused no mortality or symptoms of impaired growth against any of the insects tested, suggesting that these species are outside the protein’s target range or that the protein may not, in fact, be toxic. While the use of the polymerase chain reaction has allowed a significant increase in the number of Bt insecticidal genes characterized to date, novel NGS technologies promise a much faster, cheaper and efficient screening of Bt pesticidal proteins.

  15. Optimization of physical factors affecting the production of thermo-stable organic solvent-tolerant protease from a newly isolated halo tolerant Bacillus subtilis strain Rand

    Directory of Open Access Journals (Sweden)

    Salleh Abu

    2009-04-01

    Full Text Available Abstract Background Many researchers have reported on the optimization of protease production; nevertheless, only a few have reported on the optimization of the production of organic solvent-tolerant proteases. Ironically, none has reported on thermostable organic solvent-tolerant protease to date. The aim of this study was to isolate the thermostable organic solvent-tolerant protease and identify the culture conditions which support its production. The bacteria of genus Bacillus are active producers of extra-cellular proteases, and the thermostability of enzyme production by Bacillus species has been well-studied by a number of researchers. In the present study, the Bacillus subtilis strain Rand was isolated from the contaminated soil found in Port Dickson, Malaysia. Results A thermostable organic solvent-tolerant protease producer had been identified as Bacillus subtilis strain Rand, based on the 16S rRNA analysis conducted, as well as the morphological characteristics and biochemical properties. The production of the thermostable organic solvent-tolerant protease was optimized by varying various physical culture conditions. Inoculation with 5.0% (v/v of (AB600 = 0.5 inoculum size, in a culture medium (pH 7.0 and incubated for 24 h at 37°C with 200 rpm shaking, was the best culture condition which resulted in the maximum growth and production of protease (444.7 U/ml; 4042.4 U/mg. The Rand protease was not only stable in the presence of organic solvents, but it also exhibited a higher activity than in the absence of organic solvent, except for pyridine which inhibited the protease activity. The enzyme retained 100, 99 and 80% of its initial activity, after the heat treatment for 30 min at 50, 55, and 60°C, respectively. Conclusion Strain Rand has been found to be able to secrete extra-cellular thermostable organic solvent-tolerant protease into the culture medium. The protease exhibited a remarkable stability towards temperature and organic

  16. Survival and conjugal transfer between Bacillus thuringiensis strains in aquatic environment Sobrevivência e conjugação de Bacillus thuringiensis em ambiente aquático

    Directory of Open Access Journals (Sweden)

    Luciana Furlaneto

    2000-10-01

    Full Text Available Field and laboratory studies were conducted to assess the survival of cells and spores and plasmid transfer between Bacillus thuringienis strains in aquatic environment. Results indicated that cells and spores of B. thuringiensis can survive for 10 days in water, without altering their number. The sporulation process began after 12-15 hours of inoculation of water. B. thuringiensis was able to transfer conjugative plasmids in the aquatic environment.O presente trabalho é um estudo sobre a sobrevivência e a conjugação de linhagens de Bacillus thuringiensis em água. Os experimentos conduzidos no laboratório mostram que as células e os esporos de B. thuringiensis podem persistir pelo menos 10 dias na água. A esporulação inicia-se 12-15 horas após a inoculação. O processo de conjugação foi demonstrado em diferentes ambientes aquáticos, tanto em condições de laboratório quanto no meio ambiente.

  17. Importance of the Long-Chain Fatty Acid Beta-Hydroxylating Cytochrome P450 Enzyme YbdT for Lipopeptide Biosynthesis in Bacillus subtilis Strain OKB105

    Science.gov (United States)

    Youssef, Noha H.; Wofford, Neil; McInerney, Michael J.

    2011-01-01

    Bacillus species produce extracellular, surface-active lipopeptides such as surfactin that have wide applications in industry and medicine. The steps involved in the synthesis of 3-hydroxyacyl-coenzyme A (CoA) substrates needed for surfactin biosynthesis are not understood. Cell-free extracts of Bacillus subtilis strain OKB105 synthesized lipopeptide biosurfactants in presence of l-amino acids, myristic acid, coenzyme A, ATP, and H2O2, which suggested that 3-hydroxylation occurs prior to CoA ligation of the long chain fatty acids (LCFAs). We hypothesized that YbdT, a cytochrome P450 enzyme known to beta-hydroxylate LCFAs, functions to form 3-hydroxy fatty acids for lipopeptide biosynthesis. An in-frame mutation of ybdT was constructed and the resulting mutant strain (NHY1) produced predominantly non-hydroxylated lipopeptide with diminished biosurfactant and beta-hemolytic activities. Mass spectrometry showed that 95.6% of the fatty acids in the NHY1 biosurfactant were non-hydroxylated compared to only ∼61% in the OKB105 biosurfactant. Cell-free extracts of the NHY1 synthesized surfactin containing 3-hydroxymyristic acid from 3-hydroxymyristoyl-CoA at a specific activity similar to that of the wild type (17 ± 2 versus 17.4 ± 6 ng biosurfactant min−1·ng·protein−1, respectively). These results showed that the mutation did not affect any function needed to synthesize surfactin once the 3-hydroxyacyl-CoA substrate was formed and that YbdT functions to supply 3-hydroxy fatty acid for surfactin biosynthesis. The fact that YbdT is a peroxidase could explain why biosurfactant production is rarely observed in anaerobically grown Bacillus species. Manipulation of LCFA specificity of YbdT could provide a new route to produce biosurfactants with activities tailored to specific functions. PMID:21673922

  18. Importance of the Long-Chain Fatty Acid Beta-Hydroxylating Cytochrome P450 Enzyme YbdT for Lipopeptide Biosynthesis in Bacillus subtilis Strain OKB105

    Directory of Open Access Journals (Sweden)

    Michael J. McInerney

    2011-03-01

    Full Text Available Bacillus species produce extracellular, surface-active lipopeptides such as surfactin that have wide applications in industry and medicine. The steps involved in the synthesis of 3-hydroxyacyl-coenzyme A (CoA substrates needed for surfactin biosynthesis are not understood. Cell-free extracts of Bacillus subtilis strain OKB105 synthesized lipopeptide biosurfactants in presence of L-amino acids, myristic acid, coenzyme A, ATP, and H2O2, which suggested that 3-hydroxylation occurs prior to CoA ligation of the long chain fatty acids (LCFAs. We hypothesized that YbdT, a cytochrome P450 enzyme known to beta-hydroxylate LCFAs, functions to form 3-hydroxy fatty acids for lipopeptide biosynthesis. An in-frame mutation of ybdT was constructed and the resulting mutant strain (NHY1 produced predominantly non-hydroxylated lipopeptide with diminished biosurfactant and beta-hemolytic activities. Mass spectrometry showed that 95.6% of the fatty acids in the NHY1 biosurfactant were non-hydroxylated compared to only ~61% in the OKB105 biosurfactant. Cell-free extracts of the NHY1 synthesized surfactin containing 3-hydroxymyristic acid from 3-hydroxymyristoyl-CoA at a specific activity similar to that of the wild type (17 ± 2 versus 17.4 ± 6 ng biosurfactant min−1·ng·protein−1, respectively. These results showed that the mutation did not affect any function needed to synthesize surfactin once the 3-hydroxyacyl-CoA substrate was formed and that YbdT functions to supply 3-hydroxy fatty acid for surfactin biosynthesis. The fact that YbdT is a peroxidase could explain why biosurfactant production is rarely observed in anaerobically grown Bacillus species. Manipulation of LCFA specificity of YbdT could provide a new route to produce biosurfactants with activities tailored to specific functions.

  19. Production and Accumulation of Xylooligosaccharides with Long Chains by Growing Culture and Xylanase of a Mutant Strain of Bacillus pumilus X-6-19

    Institute of Scientific and Technical Information of China (English)

    Qingzhu Yuan; Tsuyoshi Adachi; Shinji Takenaka; Shuichiro Murakami; Machiko Tanaka; Kenji Aoki

    2008-01-01

    Bacillus pumilus X-6-9 isolated from soil and subsequently identified, produced xylooligosacchatides with long chainsfrom xylan and accumulated them in the culture. By improving the culture conditions and mutating the bacterium, a 3.2-fold increasein the production of the xylooligosaccharides was established, when compared to the original culture conditions of B. pumilus X-6-19.The addition of D-glucose to the culture of the mutant swain U-3 of B. pumilus X-6-9 repressed the synthesis of β-xylosidase, but notxylanase. Thus, it was revealed that strain U-3 was a good organism for the production and accumulation of xylooligosaccharideswith long chains from xylan by a microbial culture. Xylanase produced by strain U-3 was purified to homogeneity and characterized.The hydrolyzates generated by the purified xylanase contained xylobiose, xylotrinse, xylotewaose, and xylopentaose, but not xylose.

  20. Detection and identification of vegetative insecticidal proteins vip3 genes of Bacillus thuringiensis strains using polymerase chain reaction-high resolution melt analysis.

    Science.gov (United States)

    Li, Haitao; Shu, Changlong; He, Xiaoming; Gao, JiGuo; Liu, Rongmei; Huang, Dafang

    2012-05-01

    In this study, vegetative insecticidal proteins vip3 genes from Bacillus thuringiensis strains were detected based on polymerase chain reaction-high resolution melt (PCR-HRM) analysis. A pair of primers was designed according to the conservative sequences in 150 bp region of the known vip3 subfamily. The 150 bp regions of difference vip3 genes have only a few nucleotide difference vip3 genes were detected in 8 of 11 standard B. thuringiensis strains, and vip3Aa genes, vip3Af genes and vip3Ba gene can be distinguished as different melting curves by this method. The results demonstrate the utility of the HRM assay for mutant screening using vip3 gene. The PCR-HRM method may be a valuable and reliable tool for specific detection and identification of vip3 genes.

  1. Isolation of proline-based cyclic dipeptides from Bacillus sp. N strain associated with rhabditid [corrected] entomopathogenic nematode and its antimicrobial properties.

    Science.gov (United States)

    Kumar, Nishanth; Mohandas, C; Nambisan, Bala; Kumar, D R Soban; Lankalapalli, Ravi S

    2013-02-01

    Entomopathogenic nematodes (EPN) are well-known as biological control agents and are found to have associated bacteria which can produce a wide range of bioactive secondary metabolites. We report herewith isolation of six proline containing cyclic dipeptides cyclo(D-Pro-L-Leu), cyclo(L-Pro-L-Met), cyclo(D-Pro-L-Phe), cyclo(L-Pro-L-Phe), cyclo(L-Pro-L-Tyr) and cyclo(L-Pro-D-Tyr) from ethyl acetate extract of the Luria Broth (LB) cell free culture filtrate of Bacillus sp. strain N associated with a new EPN Rhabditis sp. from sweet potato weevil grubs collected from Central Tuber Crops Research Institute farm. Antimicrobial studies of these 2,5-diketopiperazines (DKPs) against both medicinally and agriculturally important bacterium and fungi showed potent inhibitory values in the range of μg/mL. Cyclic dipeptides showed significantly higher activity than the commercial fungicide bavistin against agriculturally important fungi, viz., Fusarium oxysporum, Rhizoctonia solani, and Pencillium expansum. The highest activity of 2 μg/mL by cyclo(L-Pro-L-Phe) was recorded against P. expansum, a plant pathogen responsible for causing post harvest decay of stored apples and oranges. To our knowledge, this is the first report on the isolation of these DKPs from Rhabditis EPN bacterial strain Bacillus sp.

  2. community Participation for culturing Local Strain of bacillus Thuringiensis H-14 in coconut Medium to controlanopheles Sundaicus Larvae at kampung Laut, cilacap regency

    Directory of Open Access Journals (Sweden)

    Wiwik Trapsilowati

    2015-01-01

    Full Text Available background: Community participation was necessary for culturinglocal strain of Bacillus thuringiensis H-14 in coconut medium and adoption this bioinsecticide to control Anopheles sundaicus. This study aimed to evaluate community participation for culturing local strain of B. thuringiensis H-14 using coconut media and spreading the results of inoculation against Anopheles sundaicus larvae. Methods: The study design was a quasi-experiment. Reseach was conducted in February-November 2007 in Kampung Laut, Cilacap. Samples were 30 person which were choosed purposively. The sample criterias were at least 15 years of age and have a pond. Community participation was evaluated for 6 times using checklist. results: Results showed that all respondents (> 80% could have on every element of 9 elements properly. Ninety percent of respondents could prepare coconut independently and made local strains of B. thuringiensis H-14 inoculation in coconuts properly (96. 67%. The density of An. sundaicuslarvae was decreased by 80–100% after 1 day spreading with culture of local strain of B. thuringiensis H-14. conclusion: Communities could participate independently for culturinglocal strain of B. thuringiensis H-14 in coconuts to control An. sundaicus larvae. The inoculation capable todecrease An. sundaicus larvae density 80–100% after 1 day. Further research was recomended to calculate cost benefit and cost effectiveness.

  3. Enterotoxigenic profiles and polymerase chain reaction detection of Bacillus cereus group cells and B. cereus strains from foods and food-borne outbreaks.

    Science.gov (United States)

    Hsieh, Y M; Sheu, S J; Chen, Y L; Tsen, H Y

    1999-10-01

    Bacillus cereus is one of the important food pathogens. Since B. cereus group cells, such as B. cereus, B. thuringiensis, B. anthracis and B. mycoides, share many phenotypical properties and a high level of chromosomal sequence similarity, it is interesting to investigate the virulence profiles for B. cereus group cells, including B. cereus strains isolated from foods and samples associated with food-poisoning outbreaks. For this investigation, the presence of enterotoxin genes, such as those of haemolysin BL, B. cereus enterotoxin T and enterotoxin FM, were assayed by polymerase chain reaction (PCR) methods. Meanwhile, their enterotoxin activities were assayed using the BCET-RPLA kit, haemolytic patterns on sheep blood agar and their cytotoxicity to Chinese hamster ovary (CHO) cells. Results showed that there were 12 enterotoxigenic profiles for the 98 B. cereus group strains collected. In addition, if any of the three types of enterotoxins was present in the B. cereus group cells, these cells were shown to be cytotoxic to the CHO cells. Similar enterotoxigenic profiles could be found among strains of B. cereus, B. mycoides and B. thuringiensis. Thus, all B. cereus group strains may be potentially toxigenic and the detection of these cells in foods is important. We thus designed PCR primers, termed Ph1/Ph2, from the sphingomyelinase gene of B. cereus cells. These primers were specific for all B. cereus group strains and could be used for the detection of B. cereus cells contaminated in food samples.

  4. Isolation and identification of the antagonistic strain DM-54 of Bacillus amyloliquefacien against Verticillium dahliae, and optimization of antifungal protein producing conditions

    Institute of Scientific and Technical Information of China (English)

    Ying ZHU; Shuna LI; Hongshui YUAN; Xiaojun GUO; Baocheng ZHU

    2009-01-01

    The strains capable of resistance against Verticillium dahliae Kleb were isolated and screened from the soils of cotton fields from several different provinces in China. A strain, coded DM-54, with a rather high antagonistic activity was obtained. Its morphological characteristics, physiological and biochemical properties and a 16 S rDNA sequence of this strain were further studied. The DM-54 strain was finally identified as a kind of Bacillus amyloliquefacien. Through a single factor experiment and an orthogonal experiment, the optimal shaking flask fermentation condition of strain DM-54 was found to be: media composed of 5% dextrin, 3% soy peptone, 0.02% MgSO4 0.01% CaCl2, initial pH 7.0 and 10% inoculum volume, media volume 30/250 (mL/mL), fermentation temperature at 32℃, rotating speed activity was distinguished to be elevated, at about 39.9%. Our research offers an effective means for the massive production of antagonistic proteins.

  5. Taxonomic Identity Resolution of Highly Phylogenetically Related Strains and Selection of Phylogenetic Markers by Using Genome-Scale Methods: The Bacillus pumilus Group Case

    Science.gov (United States)

    Espariz, Martín; Zuljan, Federico A.; Esteban, Luis; Magni, Christian

    2016-01-01

    Bacillus pumilus group strains have been studied due their agronomic, biotechnological or pharmaceutical potential. Classifying strains of this taxonomic group at species level is a challenging procedure since it is composed of seven species that share among them over 99.5% of 16S rRNA gene identity. In this study, first, a whole-genome in silico approach was used to accurately demarcate B. pumilus group strains, as a case of highly phylogenetically related taxa, at the species level. In order to achieve that and consequently to validate or correct taxonomic identities of genomes in public databases, an average nucleotide identity correlation, a core-based phylogenomic and a gene function repertory analyses were performed. Eventually, more than 50% such genomes were found to be misclassified. Hierarchical clustering of gene functional repertoires was also used to infer ecotypes among B. pumilus group species. Furthermore, for the first time the machine-learning algorithm Random Forest was used to rank genes in order of their importance for species classification. We found that ybbP, a gene involved in the synthesis of cyclic di-AMP, was the most important gene for accurately predicting species identity among B. pumilus group strains. Finally, principal component analysis was used to classify strains based on the distances between their ybbP genes. The methodologies described could be utilized more broadly to identify other highly phylogenetically related species in metagenomic or epidemiological assessments. PMID:27658251

  6. Antagonistic Activities of Bacillus spp. Strains Isolated from Tidal Flat Sediment Towards Anthracnose Pathogens Colletotrichum acutatum and C. gloeosporioides in South Korea

    Directory of Open Access Journals (Sweden)

    Joon-Hee Han

    2015-06-01

    Full Text Available Anthracnose is a fungal disease caused by Colletotrichum species that is detrimental to numerous plant species. Anthracnose control with fungicides has both human health and environmental safety implications. Despite increasing public concerns, fungicide use will continue in the absence of viable alternatives. There have been relatively less efforts to search antagonistic bacteria from mudflats harboring microbial diversity. A total of 420 bacterial strains were isolated from mudflats near the western sea of South Korea. Five bacterial strains, LB01, LB14, HM03, HM17, and LB15, were characterized as having antifungal properties in the presence of C. acutatum and C. gloeosporioides. The three Bacillus atrophaeus strains, LB14, HM03, and HM17, produced large quantities of chitinase and protease enzymes, whereas the B. amyloliquefaciens strain LB01 produced protease and cellulase enzymes. Two important antagonistic traits, siderophore production and solubilization of insoluble phosphate, were observed in the three B. atrophaeus strains. Analyses of disease suppression revealed that LB14 was most effective for suppressing the incidence of anthracnose symptoms on pepper fruits. LB14 produced antagonistic compounds and suppressed conidial germination of C. acutatum and C. gloeosporioides. The results from the present study will provide a basis for developing a reliable alternative to fungicides for anthracnose control.

  7. Whole genomic sequence analysis of Bacillus infantis: defining the genetic blueprint of strain NRRL B-14911, an emerging cardiopathogenic microbe

    Science.gov (United States)

    Background: We recently reported the identification of Bacillus sp. NRRL B-14911 that induces heart autoimmunity by generating cardiac-reactive T cells through molecular mimicry. This marine bacterium was originally isolated from the Gulf of Mexico, but no associations with human diseases were rep...

  8. The introduction of integrated pest management in the Ethiopian horticultural sector : Bacillus thuringiensis strains and its toxicity

    NARCIS (Netherlands)

    Belder, den E.; Elderson, J.

    2012-01-01

    1 Introduction As hazards of conventional broad acting pesticides are documented, researchers, poli cymakers and growers look for pesticides that are toxic only to the target pest, have no impact on other such as beneficial species, and have fewer environmental effects. Bacillus thuringiensis (Bt) i

  9. Draft Genome Sequence of Bacillus mycoides M2E15, a Strain Isolated from the Endosphere of Potato

    NARCIS (Netherlands)

    Yi, Yanglei; de Jong, Anne; Spoelder, Jan; Elzenga, J Theo M; van Elsas, Jan Dirk; Kuipers, Oscar P

    2016-01-01

    We present the draft genome sequence of Bacillus mycoides M2E15, a bacterium isolated from potato endosphere. Analysis of the 6.08-Mbp draft genome sequence identified 6,386 protein-encoding sequences, including potential plant growth promoting genes. Specifically, genes for proteins involved in pho

  10. Sporulation environment of emetic toxin-producing Bacillus cereus strains determines spore size, heat resistance and germination capacity

    NARCIS (Netherlands)

    Voort, van der M.; Abee, T.

    2013-01-01

    Aim Heat resistance, germination and outgrowth capacity of Bacillus cereus spores in processed foods are major factors in causing the emetic type of gastrointestinal disease. In this study, we aim to identify the impact of different sporulation conditions on spore properties of emetic toxin-producin

  11. 1株产聚-β-羟基丁酸芽胞杆菌的分离、筛选与鉴定%Isolation, Screening and Identification of A Poly-β-Hydroxybutyrate Yielding Bacillus Strain

    Institute of Scientific and Technical Information of China (English)

    董夏梦; 宋迤明; 蒋冬花; 蓝丽精; 蔡琪敏

    2011-01-01

    A Bacillus strain PX-95 producing poly-p-hydroxybutyrale (PHB) was isolated and purified from a peanut field soil, Jinhua, Zhejiang Province yielding PHB at 135. 81 mg/L. According to its morphological and physiological features, it was preliminarily identified as a member of Bacillus cereus group. Analysis of its 16S rDNA sequence showed strain PX-95 had a highly homology to both Bacillus tktiringiensis and Bacillus cereus. Finally it was characterized as Bacillus thuringiensis adopting amplification method of gyr B (DNA gyrase B subunit) gene.%从浙江省金华市花生地土样中筛选、分离纯化得到1株产聚-β-羟基丁酸编号为PX-95的芽胞杆菌,PHB产量为135.81 mg/L.根据形态特征、生理生化特征初步鉴定为芽胞菌属蜡样芽胞杆菌群,16S rDNA序列分析显示PX-95菌株与苏云金芽胞杆菌以及蜡样芽胞杆菌均具有高度同源性,最后采用扩增gyr B(DNA促旋酶B亚单位)基因的方法将其鉴定为苏云金芽胞杆菌(Bacillus thuringiensis ).

  12. Optimization of spore producing fermentation conditions of probiotic strain bacillus velezensis Z-27 from pig%猪源益生菌株Bacillus velezensisZ-27产芽孢发酵条件优化

    Institute of Scientific and Technical Information of China (English)

    王振海; 姜军坡; 王选; 王世英; 朱宝成

    2011-01-01

    Spore producing fermentation conditions of probiotic strain bacillus velezensis Z-27 from pig were optimized. With spore yield and biomass as main indicators, single factor test and orthogonal test were done to determine the suitable conditions for fermentation in shake flask. Results showed that biomass was 2.33 x1010 cfu/ml and spore yield was 96.0% at the optimal condition that corn meal was 1%, soybean meal 1.5%, MnSO4 0.03%,, NaH2PO4·2H2O 0.2%, Na2HPO4·2H2O 0.4%, initial pH 7.0, media volume 50 ml/250 ml, inoculation volume 2%, rotation speed 220 r/min, cultured for 24 h at 37 °C. It laid foundation for further research and application of strain bacillus velezensis Z-27.%对猪源益生菌株Bacillus velezensis Z-27进行发酵条件优化试验.以芽孢产率和生物量为指标,利用单因素试验和正交试验确定了摇瓶发酵的最佳条件:玉米粉1%、豆饼粉1.5%、MnSO40.03%、NaH2PO4·2H2O 0.2%、Na2HPO4·2H2O 0.4%,初始pH值7.0,250 ml三角瓶装量50 ml,接种量2%,摇床转速220 r/min,发酵温度37℃.优化后发酵液的生物量为2.33×1010cfu/ml,芽孢产率达96.0%,为该菌株的进一步研究和应用奠定了基础.

  13. Discrimination and phylogenomic classification of Bacillus anthracis-cereus-thuringiensis strains based on LC-MS/MS analysis of whole cell protein digests.

    Science.gov (United States)

    Dworzanski, Jacek P; Dickinson, Danielle N; Deshpande, Samir V; Snyder, A Peter; Eckenrode, Brian A

    2010-01-01

    Modern taxonomy, diagnostics, and forensics of bacteria benefit from technologies that provide data for genome-based classification and identification of strains; however, full genome sequencing is still costly, lengthy, and labor intensive. Therefore, other methods are needed to estimate genomic relatedness among strains in an economical and timely manner. Although DNA-DNA hybridization and techniques based on genome fingerprinting or sequencing selected genes like 16S rDNA, gyrB, or rpoB are frequently used as phylogenetic markers, analyses of complete genome sequences showed that global measures of genome relatedness, such as the average genome conservation of shared genes, can provide better strain resolution and give phylogenies congruent with relatedness revealed by traditional phylogenetic markers. Bacterial genomes are characterized by a high gene density; therefore, we investigated the integration of mass spectrometry-based proteomic techniques with statistical methods for phylogenomic classification of bacterial strains. For this purpose, we used a set of well characterized Bacillus cereus group strains isolated from poisoned food to describe a method that relies on liquid chromatography-electrospray ionization-tandem mass spectrometry of tryptic peptides derived from whole cell digests. Peptides were identified and matched to a prototype database (DB) of reference bacteria with fully sequenced genomes to obtain their phylogenetic profiles. These profiles were processed for predicting genomic similarities with DB bacteria estimated by fractions of shared peptides (FSPs). FSPs served as descriptors for each food isolate and were jointly analyzed using hierarchical cluster analysis methods for revealing relatedness among investigated strains. The results showed that phylogenomic classification of tested food isolates was in consonance with results from established genomic methods, thus validating our findings. In conclusion, the proposed approach could be

  14. Effect of organic fertilizers prepared from organic waste materials on the production of antibacterial volatile organic compounds by two biocontrol Bacillus amyloliquefaciens strains.

    Science.gov (United States)

    Raza, Waseem; Wei, Zhong; Ling, Ning; Huang, Qiwei; Shen, Qirong

    2016-06-10

    Three organic fertilizers made of different animal and plant waste materials (BOFs) were evaluated for their effects on the production of antibacterial volatile organic compounds (VOCs) by two Bacillus amyloliquefaciens strains SQR-9 and T-5 against the tomato wilt pathogen Ralstonia solanacearum (RS). Both strains could produce VOCs that inhibited the growth and virulence traits of RS; however, in the presence of BOFs, the production of antibacterial VOCs was significantly increased. The maximum inhibition of growth and virulence traits of RS by VOCs of T-5 and SQR-9 was determined at 1.5% BOF2 and 2% BOF3, respectively. In case of strain T-5, 2-nonanone, nonanal, xylene, benzothiazole, and butylated hydroxy toluene and in case of strain SQR-9, 2-nonanone, nonanal, xylene and 2-undecanone were the main antibacterial VOCs whose production was increased in the presence of BOFs. The results of this study reveal another significance of using organic fertilizers to improve the antagonistic activity of biocontrol agents against phytopathogens.

  15. Studies about Bacillus thuringiensis Strain Containing New cry Gene %一株含cry新基因的苏云金杆菌菌株的研究

    Institute of Scientific and Technical Information of China (English)

    宋萍; 郭丽伟; 苏俊平; 王勤英

    2011-01-01

    A high toxicity Bacillus thuringiensis (Bt)strain XPZ-9 against Lepidoptera pests was isolated and studied further. The strain formed the bipyramidal crystal and spherical crystal observed by optical microscope. SDSPAGE analysis revealed molecular weight of insecticidal crystal protein in this strain was 70 kDa. By using of PCRRFLP identification method,new crylAh gene was found in this strain. The bioassay results indicated Bt XPZ-9 toxin protein had high insecticidal activity against several Lepidoptera pests,such as 2 instar larvae of Helicoverpa armigera,Plutella xylostella, Ostrinia furnacalis, Hyphantria cunea.%苏云金杆菌XPZ-9菌株是从土壤中分离的对鳞翅目幼虫有特异杀虫活性的新菌株.光学显微镜下观察到该菌株可产生菱形和圆形伴胞晶体,SDS-PAGE 检测表达的主要蛋白条带分子量约为70 kDa.PCR-RFLP分析结果表明该菌株含有新基因cry1Ah.生物活性测定表明该菌株对鳞翅目害虫棉铃虫、小菜蛾、亚洲玉米螟和美国白蛾的二龄幼虫都具有较强的杀虫活性.

  16. A novel keratinase from Bacillus tequilensis strain Q7 with promising potential for the leather bating process.

    Science.gov (United States)

    Zaraî Jaouadi, Nadia; Rekik, Hatem; Ben Elhoul, Mouna; Zohra Rahem, Fatma; Hila, Chiraz Gorgi; Ben Aicha, Houda Slimene; Badis, Abdelmalek; Toumi, Abdessatar; Bejar, Samir; Jaouadi, Bassem

    2015-08-01

    The present paper reports on the purification and characterization of an extracellular keratinase (KERQ7) newly purified from Bacillus tequilensis Q7. Pure protein was obtained after ammonium sulfate fractionation (30-60%), followed by Mono S Sepharose cation-exchange chromatography. MALDI-TOF/MS analysis revealed that the purified enzyme was a monomer with a molecular mass of 28,355.07-Da. The sequence of the 21 N-terminal residues of KERQ7 showed high homology with those of Bacillus keratinases. Optimal activity was achieved at pH 7 and 30°C. KERQ7 was completely inhibited by PMSF and DFP, which suggests that it belongs to the serine keratinase family. KERQ7 displayed higher levels of hydrolysis and catalytic efficiency than Basozym(®) CS 10, Koropon(®) SC 5K, and Pyrase(®) 250 MP. The kerQ7 gene encoding KERQ7 was cloned, sequenced, and expressed in Escherichia coli BL21(DE3)pLysS. The biochemical properties of the extracellular purified recombinant enzyme (rKERQ7) were similar to those of native KERQ7. The deduced amino acid sequence showed strong homology with other Bacillus keratinases. The highest sequence identity value (97%) was obtained with KERUS from Brevibacillus brevis US575, with only 7 aa of difference. These properties make KERQ7 a potential promising and eco-friendly enzymatically enhanced process for animal hide bating in the leather processing industry.

  17. Development of a powder formulation based on Bacillus cereus sensu lato strain B25 spores for biological control of Fusarium verticillioides in maize plants.

    Science.gov (United States)

    Martínez-Álvarez, Juan C; Castro-Martínez, Claudia; Sánchez-Peña, Pedro; Gutiérrez-Dorado, Roberto; Maldonado-Mendoza, Ignacio E

    2016-05-01

    Maize is an economically important crop in northern Mexico. Different fungi cause ear and root rot in maize, including Fusarium verticillioides (Sacc.) Nirenberg. Crop management of this pathogen with chemical fungicides has been difficult. By contrast, the recent use of novel biocontrol strategies, such as seed bacterization with Bacillus cereus sensu lato strain B25, has been effective in field trials. These approaches are not without their problems, since insufficient formulation technology, between other factors, can limit success of biocontrol agents. In response to these drawbacks, we have developed a powder formulation based on Bacillus B25 spores and evaluated some of its characteristics, including shelf life and efficacy against F. verticillioides, in vitro and in maize plants. A talc-based powder formulation containing 1 × 10(9) c.f.u. g(-1) was obtained and evaluated for seed adherence ability, seed germination effect, shelf life and antagonism against F. verticillioides in in vitro and in planta assays. Seed adherence of viable bacterial spores ranged from 1.0 to 1.41 × 10(7) c.f.u. g(-1). Bacteria did not display negative effects on seed germination. Spore viability for the powder formulation slowly decreased over time, and was 53 % after 360 days of storage at room temperature. This formulation was capable of controlling F. verticillioides in greenhouse assays, as well as eight other maize phytopathogenic fungi in vitro. The results suggest that a talc-based powder formulation of Bacillus B25 spores may be sufficient to produce inoculum for biocontrol of maize ear and root rots caused by F. verticillioides.

  18. 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

  19. Concomitant production of two proteases and alpha-amylase by a novel strain of Bacillus subtilis in a microprocessor controlled bioreactor

    Directory of Open Access Journals (Sweden)

    Hamid Mukhtar

    2012-09-01

    Full Text Available We describe the simultaneous production of Bacillus subtilis based proteases and alpha amylase using a computer controlled laboratory scale 7.5 L batch bioreactor. The present strain is the first to be reported that concomitantly produces these two industrially important enzymes. The growth and sporulation of Bacillus subtilis was monitored and maximum production of alkaline protease and alpha amylase was found to coincide with maximum sporulation. Two types of proteases were detected in the fermentation broth; a neutral and an alkaline protease most active in a pH range of 7.0-8.0 and 8.0-10, respectively. Maximum production of proteases was observed at an incubation temperature of 37ºC while that of alpha amylase was observed at 40ºC. The optimum aeration and agitation levels for protease production were 0.6 L/L/min and 200rpm, respectively, and for alpha amylase were 0.6 L/L/min and 150 rpm. The kinetic parameters Yp/x and qp were also found to be significant at the given fermentation conditions.

  20. Adverse reactions to the Bacillus Calmette-Guérin (BCG) vaccine in new-born infants-an evaluation of the Danish strain 1331 SSI in a randomized clinical trial

    DEFF Research Database (Denmark)

    Nørrelykke Nissen, Thomas; Birk, Nina Marie; Kjærgaard, Jesper;

    2016-01-01

    OBJECTIVE: To evaluate adverse reactions of the Bacillus Calmette-Guérin (BCG) Statens Serum Institut (SSI) (Danish strain 1331) used as intervention in a randomized clinical trial. DESIGN: A randomized clinical multicenter trial, The Danish Calmette Study, randomizing newborns to BCG or no inter...

  1. Assessment of Bacillus thuringiensis subsp. israelensis Strain AM65-52 aqueous suspension for the control of black fly populations, (Simulium spp. in northern Italy

    Directory of Open Access Journals (Sweden)

    P. Radeghieri

    2013-12-01

    Full Text Available Four field trials were conducted in Mantua province in northern Italy to evaluate the effect of the Bacillus thuringiensis subsp. israelensis Strain AM65-52 based VectoBac® 12 AS larvicide on black flies. Our objective was to examine and compare the efficacy of different concentrations of this microbial larvicide in small plain streams. Larvicidal activity was exhibited in all trials. VectoBac® 12 AS at 2.5 ppm/min rate generated larval mortality from the treatment point to 500 meters downstream. Larvae mortality (over 80% was observed for 3500 meters downstream of the application site when a concentration of 40 ppm/min was applied.

  2. Effect of glutathione L-cystein and L-djenkolic acid in the synthesis and mutagenicity of azide metabolite in Bacillus subtilis ATCC 6633 strain.

    Science.gov (United States)

    Elbetieha, A; Owais, W M; Saadoun, I; Hussein, E

    1999-10-01

    The Bacillus subtilis ATCC 6633 strain synthesizes a mutagenic metabolite from sodium azide and O-acetylserine. Mutagenicity of azide was decreased in growth media containing 10(-4) M glutathione, L-cysteine or L-djenkolic acid whereas dithiothritol (DTT) added at the same concentration did not reduce the mutagenicity of azide. Likewise, glutathione, L-cysteine, L-djenkolic acid, and DTT were found to have no effect in reducing the mutagenicity of the in vitro produced metabolite using bacterial cell-free extract. These results suggest that O-acetyl-serine sulfhydrylase catalyzes the reaction of azide and O-acetylserine to form a mutagenic metabolite, which is ninhydrin positive and migrates in TLC to an Rf value similar to that of azidoalanine in both acidic and basic solvent systems.

  3. Bme585 I [5'-CCCGC(4/6)-3'], a new isoschizomer of restriction endonuclease Fau I, isolated from a strain of Bacillus mesentericus.

    Science.gov (United States)

    Davalieva, Katarina; Ziberovski, Jugoslav; Efremov, Georgi D

    2004-01-01

    Bme585 I is a new member of the restriction endonuclease type IIS family. It was partially purified from the heterothrophic, mesophilic bacterial strain Bacillus mesentericus 585 by ammonium sulphate precipitation and phosphocellulose column chromatography. Bme585 I is a monomeric protein with a molecular mass of 62 kD. The enzyme is active over a broad pH range from 7.0 to 8.8, has a temperature optimum of 37 degrees C and tolerance of NaCl in reaction buffer from 0 to 400 mM. Bme585 I recognizes the asymmetric sequence 5'-CCCGC(4/6)-3' and is therefore an isoschizomer of restriction endonuclease Fau I.

  4. 一株土壤源高产纤维素酶芽孢杆菌的分离与鉴定%Isolation and Identification of a High Yield Cellulase Bacillus Strain from Soil

    Institute of Scientific and Technical Information of China (English)

    董鹤娟; 丁轲; 恒子钤; 贾艳艳; 彭春平; 罗伟光; 李旺

    2013-01-01

    In order to obtain the high yield cellulase bacillus,58 soil samples from different areas in Henan province were collected,the bacillus producing cellulase was isolated by Congo red medium,and the cellulase activity was detected by 3,5-dinitro salicylic acid method.The high cellulase producing bacillus were screened and identified by the colony morphology,microscopic morphology in combination with the physiological and biochemical feature and 16S rDNA sequence.The results showed that the strain B.LY02 with the highest cellulase activity of 0.5351 U · mL-1 was obtained,and it was short rod,forming spores,gram strain positive,and had the ability of fermenting many sugars.The 16S rDNA sequence homology comparative analysis showed that it belonged to the genus Bacillus and most closery related to Bacillus licheniformis strain CICC10095 with 96.5% sequence simmarity.So the strain Bacillus LY02 was identified to be Bacillus licheniformis.%为了获得产纤维素酶的芽孢杆菌,从河南省不同地方采集玉米地、秸杆垛的土壤样品58份,利用刚果红平板法分离产纤维素酶的芽孢杆菌,采用3,5—二硝基水杨酸法测定纤维素酶活,结合茵落特征、显微形态、生理生化试验和16S rDNA序列进行鉴定.结果表明,从样品中分离出的42株产纤维素酶菌株中筛选出一株产纤维素酶较高的菌株B.LY02,纤维素酶活力可达0.5351 U/mL.该菌株呈短杆状,革兰氏染色阳性,能形成芽孢.基于16S rDNA序列同源性比较分析表明该菌株与Bacillus licheni ormis strain CICC10095的亲缘关系最近,基因序列的同源性为96.5%,因此鉴定该菌株为地衣芽孢杆菌.

  5. Response of tomato wilt pathogen Ralstonia solanacearum to the volatile organic compounds produced by a biocontrol strain Bacillus amyloliquefaciens SQR-9.

    Science.gov (United States)

    Raza, Waseem; Ling, Ning; Yang, Liudong; Huang, Qiwei; Shen, Qirong

    2016-04-22

    It is important to study the response of plant pathogens to the antibiosis traits of biocontrol microbes to design the efficient biocontrol strategies. In this study, we evaluated the role of volatile organic compounds (VOCs) produced by a biocontrol strain Bacillus amyloliquefaciens SQR-9 on the growth and virulence traits of tomato wilt pathogen Ralstonia solanacearum (RS). The VOCs of SQR-9 significantly inhibited the growth of RS on agar medium and in soil. In addition, the VOCs significantly inhibited the motility traits, production of antioxidant enzymes and exopolysaccharides, biofilm formation and tomato root colonization by RS. The strain SQR-9 produced 22 VOCs, but only nine VOCs showed 1-11% antibacterial activity against RS in their corresponding amounts; however, the consortium of all VOCs showed 70% growth inhibition of RS. The proteomics analysis showed that the VOCs of SQR-9 downregulated RS proteins related to the antioxidant activity, virulence, carbohydrate and amino acid metabolism, protein folding and translation, while the proteins involved in the ABC transporter system, amino acid synthesis, detoxification of aldehydes and ketones, methylation, protein translation and folding, and energy transfer were upregulated. This study describes the significance and effectiveness of VOCs produced by a biocontrol strain against tomato wilt pathogen.

  6. Cry-like genes, in an uncommon gene configuration, produce a crystal that localizes within the exosporium when expressed in an acrystalliferous strain of Bacillus thuringiensis.

    Science.gov (United States)

    Ammons, David; Toal, Graham; Roman, Angel; Rojas-Avelizapa, Luz I; Ventura-Suárez, Antonio; Rampersad, Joanne

    2016-02-01

    Cry proteins are pesticidal toxins produced by the bacterium Bacillus thuringiensis (Bt), which aggregate in sporulating cells to form a crystal. Except in a relatively few cases, these crystals are located outside the exosporium that surrounds the spore. Bt2-56 is a strain of Bt that has the relatively uncommon characteristic of locating its Cry protein-containing crystal within the exosporium, and in association with a long, multifiber filament. With the ultimate goal of both understanding and manipulating the localization of Cry proteins within the exosporium, we sought to identify the genes coding for the exosporium-localized Cry proteins in Bt2-56. Herein we show (i) that five cry-like genes are present in the genome of Bt2-56, (ii) that two pairs of these genes show organizational similarity to a relatively uncommon gene configuration that coexpress a cry gene along with a gene whose product aids crystal formation and (iii) that when one of these two gene pairs (cry21A-cdA) is expressed in an acrystalliferous strain of Bt, crystals are formed that localize within the exosporium. In Bt ssp. finitimus, the only other strain in which crystal localization has been studied, a Cry protein needed expression of two non-cry ORFs in order to localize within the exosporium, indicating that there are some mechanistic differences for crystal localization between Bt ssp. finitimus and Bt2-56.

  7. Potential plant growth-promoting strain Bacillus sp. SR-2-1/1 decolorized azo dyes through NADH-ubiquinone:oxidoreductase activity.

    Science.gov (United States)

    Mahmood, Faisal; Shahid, Muhammad; Hussain, Sabir; Shahzad, Tanvir; Tahir, Muhammad; Ijaz, Muhammad; Hussain, Athar; Mahmood, Khalid; Imran, Muhammad; Babar, Shahid Ali Khan

    2017-03-22

    In this study, a bacterial strain SR-2-1/1 was isolated from textile wastewater-irrigated soil for its concurrent potential of plant growth promotion and azo-dye decolorization. Analysis of 16S rRNA gene sequence confirmed its identity as Bacillus sp. The strain tolerated high concentrations (i.e. up to 1000mgL(-1)) of metals (Ni(2+), Cd(2+), Co(2+), Zn(2+), and Cr(6+)) and efficiently decolorized the azo dyes (i.e. reactive black-5, reactive red-120, direct blue-1 and congo red). It also demonstrated considerable in vitro phosphate solubilizing and 1-aminocyclopropane-1-carboxylic acid deaminase abilities at high metal and salt levels. Bioinformatics analysis of its 537bp azoreductase gene and deduced protein revealed that it decolorized azo dyes through NADH-ubiquinone:oxidoreductase enzyme activity. The deduced protein was predicted structurally and functionally different to those of its closely related database proteins. Thus, the strain SR-2-1/1 is a powerful bioinoculant for bioremediation of textile wastewater contaminated soils in addition to stimulation of plant growth.

  8. Draft Genome Sequence of Commercial Textile Dye-Decolorizing and -Degrading Bacillus subtilis Strain C3 Isolated in India

    Science.gov (United States)

    Kunadia, Khushbu; Nathani, Neelam M.; Kothari, Vishal; Kotadia, Rohit J.; Kothari, Charmy R.; Joshi, Anjali; Rank, Jalpa K.; Faldu, Priti R.; Shekar, M. Chandra; Viroja, Mitkumar J.; Patel, Priyank A.; Jadeja, Divyarajsinh; Reddy, Bhaskar; Pal Singh, Ravindra; Koringa, Prakash G.; Joshi, Chaitanya G.

    2016-01-01

    Bacillus subtilis C3, a commercial textile dye-decolorizing and -degrading bacterium, was isolated from the common effluent treatment plant (CEPT) of the Jetpur textile dyeing and printing industrial sector situated in the district of Rajkot, Gujarat, India. Here, we present the annotated 4.18-Mb draft genome sequence of B. subtilis C3, providing information about the metabolic pathways involved in decolorization and degradation of several commercial textile azo dyes. Thus, we confirm B. subtilis C3 as a potential candidate for bioremediation of textile effluents. PMID:26966205

  9. Draft Genome Sequence of Commercial Textile Dye-Decolorizing and -Degrading Bacillus subtilis Strain C3 Isolated in India.

    Science.gov (United States)

    Kunadia, Khushbu; Nathani, Neelam M; Kothari, Vishal; Kotadia, Rohit J; Kothari, Charmy R; Joshi, Anjali; Rank, Jalpa K; Faldu, Priti R; Shekar, M Chandra; Viroja, Mitkumar J; Patel, Priyank A; Jadeja, Divyarajsinh; Reddy, Bhaskar; Pal Singh, Ravindra; Koringa, Prakash G; Joshi, Chaitanya G; Kothari, Ramesh K

    2016-03-10

    Bacillus subtilis C3, a commercial textile dye-decolorizing and -degrading bacterium, was isolated from the common effluent treatment plant (CEPT) of the Jetpur textile dyeing and printing industrial sector situated in the district of Rajkot, Gujarat, India. Here, we present the annotated 4.18-Mb draft genome sequence of B. subtilis C3, providing information about the metabolic pathways involved in decolorization and degradation of several commercial textile azo dyes. Thus, we confirm B. subtilis C3 as a potential candidate for bioremediation of textile effluents.

  10. Complete genome sequence of Bacillus thuringiensis YC-10, a novel active strain against plant-parasitic nematodes.

    Science.gov (United States)

    Cheng, Feixue; Wang, Jian; Song, Zhiqiang; Cheng, Ju'e; Zhang, Deyong; Liu, Yong

    2015-09-20

    Bacillus thuringiensis is an important microbial biopesticide for controlling agricultural pests by the production of toxic parasporal crystals proteins.Here,we report the finished annotated genome sequence of B. thuringiensis YC-10,which is highly toxic to nematodes.The complete genome sequence consists of a circular chromosome and nine circular plasmids,which the biggest plasmid harbors six parasporal crystals proteins genes consisting of cry1Aa, cry1Ac, cry1Ia, cry2Aa, cry2Ab and cryB1. The crystals proteins of Cry1Ia and Cry1Aa have high nematicidal activity against Meloidogyne incognita.

  11. Isolation and characterization of a β-propeller gene containing phosphobacterium Bacillus subtilis strain KPS-11 for growth promotion of potato (Solanum tuberosum L.

    Directory of Open Access Journals (Sweden)

    Kashif eHanif

    2015-06-01

    Full Text Available Phosphate-solubilizing and phytate-mineralizing bacteria collectively termed as phosphobacteria provide a sustainable approach for managing P-deficiency in agricultural soils by supplying inexpensive phosphate to plants. A phosphobacterium Bacillus subtilis strain KPS-11 (Genbank accession no. KP006655 was isolated from potato (Solanum tuberosum L. rhizosphere and characterized for potato plant growth promoting potential. The strain utilized both Ca-phosphate and Na-phytate in vitro and produced 6.48 µg mL-1 indole-3-acetic acid in tryptophan supplemented medium. P-solubilization after 240 h was 66.4 µg mL-1 alongwith the production of 19.3 µg mL-1 gluconic acid and 5.3 µg mL-1 malic acid. The extracellular phytase activity was higher (4.3 x 10-10 kat mg-1 protein than the cell-associated phytase activity (1.6 x 10-10 kat mg-1protein. B. subtilis strain KPS-11 utilized 40 carbon sources and showed resistance against 20 chemicals in GENIII micro-plate system demonstrating its metabolic potential. Phytase-encoding gene β-propeller (BPP showed 92% amino acid similarity to BPP from B. subtilis (accession no.WP_014114128.1 and 83% structural similarity to BPP from B. subtilis (accession no 3AMR_A. Potato inoculation with B. subtilis strain KPS-11 increased the root/shoot length and root/shoot weight of potato as compared to non-inoculated control plants. Moreover, rifampicin-resistant derivative of KPS-11 were able to survive in the rhizosphere and on the roots of potato up to sixty days showing its colonization potential. The study indicates that B. subtilis strain KPS-11 can be a potential candidate for development of potato inoculum in P-deficient soils.

  12. A promising HD133-like strain of Bacillus thuringiensis with dual efficiency to the two Lepidopteran pests: Spodoptera littoralis (Noctuidae) and Ephestia kuehniella (Pyralidae).

    Science.gov (United States)

    BenFarhat-Touzri, Dalel; Driss, Fatma; Tounsi, Slim

    2016-08-01

    Isolation and identification of new strains with wide variety of target pests is an ever growing field. In this paper, a screening of 260 strains from Tunisian soil samples was conducted by dot-blot and PCR-sequencing analysis. The screening was done on the basis of the possession of cry1D-type genes and was followed by the evaluation of the insecticidal activity against Spodoptera littoralis. BLB250 strain showed an LC50 value (56.2 μg g(-1)) against S. littoralis lower than those of the two Bacillus thuringiensis reference strains HD1 and HD133. An interesting LC50 (167.6 μg g(-1)) was also recorded against Ephestia kuehniella larvae. The strain was, thus, selected because of its qualification to be highly toxic, at once, for both Lepidopteran insects. In vitro time course of proteolytic processing of BLB250 and HD133 protoxins by the gut juices from the two insect larvae revealed that the differences in toxicity against E. kuehniella are most likely attributed to differences in the efficiency of the activation of the corresponding protoxins into toxins. An activation comparative study using commercial proteases suggested that the intestinal proteases of E. kuehniella contain trypsin-like activities. With its high efficiency and toxicity against, at once, two Lepidopteran insects having different susceptibilities towards kurstaki and aizawai subspecies, BLB250 could be useful when developing more efficient and economical B. thuringiensis-based pesticides.

  13. Requirement of Simultaneous Assessment of Crystal- and Supernatant-Related Entomotoxic Activities of Bacillus thuringiensis Strains for Biocontrol-Product Development

    Science.gov (United States)

    Argôlo-Filho, Ronaldo Costa; Costa, Robson Luz; Pinheiro, Daniele Heloisa; Corrêa, Fábio Mathias; Valicente, Fernando Hercos; Pomella, Alan William Vilela; Loguercio, Leandro Lopes

    2014-01-01

    Bioinsecticides with lower concentrations of endospores/crystals and without loss of efficiency are economically advantageous for pest biocontrol. In addition to Cry proteins, other Bacillus thuringiensis (Bt) toxins in culture supernatants (SN) have biocontrol potential (e.g., Vip3A, Cry1I, Sip1), whereas others are unwanted (β-exotoxins), as they display widespread toxicity across taxa. A strain simultaneously providing distinct toxin activities in crystals and SN would be desirable for bioinsecticides development; however, strains secreting β-exotoxins should be discarded, independently of other useful entomotoxins. Entomotoxicity of crystals and SN from a Brazilian Bt tolworthi strain (Btt01) was tested against Spodoptera frugiperda to assess the potential for biocontrol-product development based on more than one type of toxin/activity. Tests showed that 107 endospores mL−1 caused >80% of larvae mortality, suggesting Btt01 may be used in similar concentrations as those of other Bt-based biopesticides. When it was applied to cornfields, a significant 60% reduction of larvae infestation was observed. However, bioassays with Btt01 SN revealed a thermostable toxic activity. Physicochemical characterization strongly suggests the presence of unwanted β-exotoxins, with isolate-specific temporal variation in its secretion. Knowledge of the temporal pattern of secretion/activity in culture for all forms of toxins produced by a single strain is required to both detect useful activities and avoid the potential lack of identification of undesirable toxins. These findings are discussed in the contexts of commercial Bt product development, advantages of multiple-activity strains, and care and handling recommended for large-scale fermentation systems. PMID:24854738

  14. Requirement of simultaneous assessment of crystal- and supernatant-related entomotoxic activities of Bacillus thuringiensis strains for biocontrol-product development.

    Science.gov (United States)

    Argôlo-Filho, Ronaldo Costa; Costa, Robson Luz; Pinheiro, Daniele Heloisa; Corrêa, Fábio Mathias; Valicente, Fernando Hercos; Pomella, Alan William Vilela; Loguercio, Leandro Lopes

    2014-05-20

    Bioinsecticides with lower concentrations of endospores/crystals and without loss of efficiency are economically advantageous for pest biocontrol. In addition to Cry proteins, other Bacillus thuringiensis (Bt) toxins in culture supernatants (SN) have biocontrol potential (e.g., Vip3A, Cry1I, Sip1), whereas others are unwanted (β-exotoxins), as they display widespread toxicity across taxa. A strain simultaneously providing distinct toxin activities in crystals and SN would be desirable for bioinsecticides development; however, strains secreting β-exotoxins should be discarded, independently of other useful entomotoxins. Entomotoxicity of crystals and SN from a Brazilian Bt tolworthi strain (Btt01) was tested against Spodoptera frugiperda to assess the potential for biocontrol-product development based on more than one type of toxin/activity. Tests showed that 10(7) endospores mL(-1) caused >80% of larvae mortality, suggesting Btt01 may be used in similar concentrations as those of other Bt-based biopesticides. When it was applied to cornfields, a significant 60% reduction of larvae infestation was observed. However, bioassays with Btt01 SN revealed a thermostable toxic activity. Physicochemical characterization strongly suggests the presence of unwanted β-exotoxins, with isolate-specific temporal variation in its secretion. Knowledge of the temporal pattern of secretion/activity in culture for all forms of toxins produced by a single strain is required to both detect useful activities and avoid the potential lack of identification of undesirable toxins. These findings are discussed in the contexts of commercial Bt product development, advantages of multiple-activity strains, and care and handling recommended for large-scale fermentation systems.

  15. Requirement of Simultaneous Assessment of Crystal- and Supernatant-Related Entomotoxic Activities of Bacillus thuringiensis Strains for Biocontrol-Product Development

    Directory of Open Access Journals (Sweden)

    Ronaldo Costa Argôlo-Filho

    2014-05-01

    Full Text Available Bioinsecticides with lower concentrations of endospores/crystals and without loss of efficiency are economically advantageous for pest biocontrol. In addition to Cry proteins, other Bacillus thuringiensis (Bt toxins in culture supernatants (SN have biocontrol potential (e.g., Vip3A, Cry1I, Sip1, whereas others are unwanted (β-exotoxins, as they display widespread toxicity across taxa. A strain simultaneously providing distinct toxin activities in crystals and SN would be desirable for bioinsecticides development; however, strains secreting β-exotoxins should be discarded, independently of other useful entomotoxins. Entomotoxicity of crystals and SN from a Brazilian Bt tolworthi strain (Btt01 was tested against Spodoptera frugiperda to assess the potential for biocontrol-product development based on more than one type of toxin/activity. Tests showed that 107 endospores mL−1 caused >80% of larvae mortality, suggesting Btt01 may be used in similar concentrations as those of other Bt-based biopesticides. When it was applied to cornfields, a significant 60% reduction of larvae infestation was observed. However, bioassays with Btt01 SN revealed a thermostable toxic activity. Physicochemical characterization strongly suggests the presence of unwanted β-exotoxins, with isolate-specific temporal variation in its secretion. Knowledge of the temporal pattern of secretion/activity in culture for all forms of toxins produced by a single strain is required to both detect useful activities and avoid the potential lack of identification of undesirable toxins. These findings are discussed in the contexts of commercial Bt product development, advantages of multiple-activity strains, and care and handling recommended for large-scale fermentation systems.

  16. [Genes of insecticidal crystal proteins with dual specificity in Bacillus thuringiensis strains, isolated in the Crimea territory].

    Science.gov (United States)

    Rymar, S Iu; Isakova, I A; Kuznietsova, L M; Kordium, V A

    2006-01-01

    The insecticidal crystal proteins of 15 B. thuringiensis strains, isolated in the Crimea territory that are toxical for some Lepidoptera and Colorado potato beetle larvae were identified by PAGE electrophoresis. Ten strains produced the crystal proteins with high molecular weight (> 120 kD). PCR with use of broad specificity primers and DNA of these B. thuringiensis strains as template demonstrated the specific PCR products (1000 bp). Amplified DNA fragments were cloned and sequenced. The nucleotide sequence analysis revealed that the genomes of ten strains of B. thuringiensis carried Cry1B genes, which are responsible for production of the insecticidal crystal proteins with dual specificity. The influence of the solubilization conditions on the structure and toxicity of Cry1B protein for Colorado potato beetle larvae was shown. The dual toxicity of studied B. thuringiensis strains is explained by the Cry1B genes presence in their genomes. These strains may be used to develop the broad specificity bioinsecticides.

  17. Thermostability enhancement and change in starch hydrolysis profile of the maltohexaose-forming amylase of Bacillus stearothermophilus US100 strain.

    Science.gov (United States)

    Ben Ali, Mamdouh; Khemakhem, Bassem; Robert, Xavier; Haser, Richard; Bejar, Samir

    2006-02-15

    The implications of Asn315 and Val450 in the atypical starch hydrolysis profile of Bacillus stearothermophilus Amy (a-amylase) US100 have been suggested previously [Ben Ali, Mhiri, Mezghani and Bejar (2001) Enzyme Microb. Tech. 28, 537-542]. In order to confirm this hypothesis, three mutants were generated. Of these two have a single mutation, N315D or V450G, whereas the third contains both mutations. Analysis of the starch breakdown-profile of these three mutants, as well as of the wild-type, allowed us to conclude that each single mutation induces a small variation in the hydrolysis product. However, the major end product produced by the double mutant shifts from maltopentaose/maltohexaose to maltose/maltotriose, confirming the involvement of these two residues in starch hydrolysis. The superimposition of AmyUS100 model with that of Bacillus licheniformis shows in AmyUS100 an additional loop containing residues Ile214 and Gly215. Remarkably, the deletion of these two residues increases the half-life at 100 degrees C from 15 min to approx. 70 min. Moreover, this engineered amylase requires less calcium, 25 p.p.m. instead of 100 p.p.m., to reach maximal thermostability.

  18. Comparison of French and Worldwide Bacillus anthracis Strains Favors a Recent, Post-Columbian Origin of the Predominant North-American Clade.

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    Gilles Vergnaud

    Full Text Available Bacillus anthracis, the highly dangerous zoonotic bacterial pathogen species is currently composed of three genetic groups, called A, B and C. Group A is represented worldwide whereas group B is present essentially in Western Europe and Southern Africa. Only three strains from group C have been reported. This knowledge is derived from the genotyping of more than 2000 strains collected worldwide. Strains from both group A and group B are present in France. Previous investigations showed that the majority of sporadic French strains belong to the so-called A.Br.011/009 group A clade and define a very remarkable polytomy with six branches. Here we explore the significance of this polytomy by comparing the French B. anthracis lineages to worldwide lineages. We take advantage of whole genome sequence data previously determined for 122 French strains and 45 strains of various origins.A total of 6690 SNPs was identified among the available dataset and used to draw the phylogeny. The phylogeny of the French B group strains which belongs to B.Br.CNEVA indicates an expansion from the south-east part of France (the Alps towards the south-west (Massif-Central and Pyrenees. The relatively small group A strains belonging to A.Br.001/002 results from at least two independent introductions. Strikingly, the data clearly demonstrates that the currently predominant B. anthracis lineage in North America, called WNA for Western North American, is derived from one branch of the A.Br.011/009 polytomy predominant in France.The present work extends the range of observed substitution rate heterogeneity within B. anthracis, in agreement with its ecology and in contrast with some other pathogens. The population structure of the six branches A.Br.011/009 polytomy identified in France, diversity of branch length, and comparison with the WNA lineage, suggests that WNA is of post-Columbian and west European origin, with France as a likely source. Furthermore, it is tempting to

  19. Screening and characterization of endophytic Bacillus and Paenibacillus strains from medicinal plant Lonicera japonica for use as potential plant growth promoters.

    Science.gov (United States)

    Zhao, Longfei; Xu, Yajun; Lai, Xin-He; Shan, Changjuan; Deng, Zhenshan; Ji, Yuliang

    2015-01-01

    strains were identified to be Paenibacillus and Bacillus strains based on the results of 16S rRNA gene sequencing analysis and their physiological and biochemical characteristics. Results indicate the promising application of endophytic bacteria to the biological control of pathogenic fungi and the improvement of wheat crop growth.

  20. Whole Genome Sequencing and Phylogenetic Analysis of a Historical Collection of Bacillus anthracis Strains from Danish Cattle

    DEFF Research Database (Denmark)

    Derzelle, Sylviane; Girault, Guillaume; Kokotovic, Branko

    2015-01-01

    of such lineage in Europe is demonstrated for the first time, filling an historical gap within the phylogeography of the lineage. Comparative genome analyses of these strains with 41 isolates from other parts of the world revealed that the two Danish A.Br.008/011 strains were related to the heroin...

  1. Histopathology and the lethal effect of Cry proteins and strains of Bacillus thuringiensis Berliner in Spodoptera frugiperda J.E. Smith Caterpillars (Lepidoptera, Noctuidae

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    N. Knaak

    Full Text Available Among the phytophagous insects which attack crops, the fall armyworm, Spodoptera frugiperda (J.E. Smith, 1797 (Lepidoptera, Noctuidae is particularly harmful in the initial growth phase of rice plants. As a potential means of controlling this pest, and considering that the entomopathogen Bacillus thuringiensis Berliner demonstrates toxicity due to synthesis of the Cry protein, the present study was undertaken to evaluate this toxic effect of B. thuringiensis thuringiensis 407 (pH 408 and B. thuringiensis kurstaki HD-73 on S. frugiperda. The following method was used. Both bacterial strains were evaluated in vitro in 1st instar S. frugiperda caterpillars, by means of histopathological assays. The Cry1Ab and Cry1Ac proteins, codified by the respective strains of B. thuringiensis, were evaluated in vivo by bioassays of 1st instar S. frugiperda caterpillars in order to determine the Mean Lethal Concentration (LC50. The results of the histopathological analysis of the midget of S. frugiperda caterpillars demonstrate that treatment with the B. thuringiensis thuringiensis strain was more efficient, because the degradations of the microvilosities started 9 hours after treatment application (HAT, while in the B. thuringiensis kurstaki the same effect was noticed only after 12 HAT. Toxicity data of the Cry1Ab and Cry1Ac proteins presented for the target-species LC50 levels of 9.29 and 1.79 μg.cm-2 respectively. The strains and proteins synthesised by B. thuringiensis thuringiensis and B. thuringiensis kurstaki are effective in controlling S. frugiperda, and may be used to produce new biopesticides or the genes may be utilised in the genetic transformation of Oryza sativa L.

  2. Histopathology and the lethal effect of Cry proteins and strains of Bacillus thuringiensis Berliner in Spodoptera frugiperda J.E. Smith Caterpillars (Lepidoptera, Noctuidae).

    Science.gov (United States)

    Knaak, N; Franz, A R; Santos, G F; Fiuza, L M

    2010-08-01

    Among the phytophagous insects which attack crops, the fall armyworm, Spodoptera frugiperda (J.E. Smith, 1797) (Lepidoptera, Noctuidae) is particularly harmful in the initial growth phase of rice plants. As a potential means of controlling this pest, and considering that the entomopathogen Bacillus thuringiensis Berliner demonstrates toxicity due to synthesis of the Cry protein, the present study was undertaken to evaluate this toxic effect of B. thuringiensis thuringiensis 407 (pH 408) and B. thuringiensis kurstaki HD-73 on S. frugiperda. The following method was used. Both bacterial strains were evaluated in vitro in 1st instar S. frugiperda caterpillars, by means of histopathological assays. The Cry1Ab and Cry1Ac proteins, codified by the respective strains of B. thuringiensis, were evaluated in vivo by bioassays of 1st instar S. frugiperda caterpillars in order to determine the Mean Lethal Concentration (LC50). The results of the histopathological analysis of the midget of S. frugiperda caterpillars demonstrate that treatment with the B. thuringiensis thuringiensis strain was more efficient, because the degradations of the microvilosities started 9 hours after treatment application (HAT), while in the B. thuringiensis kurstaki the same effect was noticed only after 12 HAT. Toxicity data of the Cry1Ab and Cry1Ac proteins presented for the target-species LC50 levels of 9.29 and 1.79 microgxcm-2 respectively. The strains and proteins synthesised by B. thuringiensis thuringiensis and B. thuringiensis kurstaki are effective in controlling S. frugiperda, and may be used to produce new biopesticides or the genes may be utilised in the genetic transformation of Oryza sativa L.

  3. Strain Improvement of Bacillus coagulans and Geobacillus stearothermophilus for Enhanced Thermostable Cellulase Production and the Effect of Different Metal Ions on Cellulase Activity

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    Vikas Sharma

    2012-11-01

    Full Text Available The current study was focused on the strain improvement of Bacillus coagulans and Geobacillus stearothermophilus for thermostable cellulase production with higher enzyme activity. For strain improvement UV radiations, NTG and Sodium azide were used as mutagenic agents.NTG was found to be best mutagenic agent among all in term of highest cellulase activity. Mutant strain C11 exhibited the highest cellulase specific activity at 45 U/mg followed by C15 (39 U/mg in case of B.coagulans while Mutant strain S18 exhibited thehighest cellulase specific activity at 69 U/mg followed by S12 (62 U/mg in case of G. stearothermophilus. Specific activity of cellulase was 92 U/mg in case of B.coagulans C11 and 118 U/mg in case of G. stearothermophilus S18. Ag+, Mg+, Se2+,Ca2+,Co2+,Mn2+,K+, Zn2+ ,Fe3+, Hg2+ and Cu2+ showed positive change in specific activity while Na+, Ni2+ negative change in specific activity of cellulase with respect to specific activity of cellulase in absence of any additive in case of B.coagulans C11 and Ag+, Mg+, Se2+,Co2+,Mn2+ andHg2+ showed positive change in specific activity, Na+, K+ showed no change in specific activity while Ca2+, Zn2+, Ni2+, Fe3+ and Cu2+ showed negative change in specific activity of cellulase with respect to specific activity of cellulase in absence of any additive in case of G. stearothermophilus S18.

  4. Genotyping of French Bacillus anthracis strains based on 31-loci multi locus VNTR analysis: epidemiology, marker evaluation, and update of the internet genotype database.

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    Simon Thierry

    Full Text Available BACKGROUND: Bacillus anthracis is known to have low genetic variability. In spite of this lack of diversity, multiple-locus variable-number tandem repeat (VNTR analysis (MLVA and single nucleotide polymorphisms (SNPs including the canonical SNPs assay (canSNP have proved to be highly effective to differentiate strains. Five different MLVA schemes based on a collection of 31 VNTR loci (MLVA8, MLVA15, MLVA20, MLVA25 and MLVA31 with increased resolving power have been described. RESULTS: MLVA31 was applied to characterize the French National Reference Laboratory collection. The total collection of 130 strains is resolved in 35 genotypes. The 119 veterinary and environmental strains collection in France were resolved into 26 genotypes belonging to three canSNP lineages and four MLVA clonal complexes (CCs with particular geographical clustering. A subset of seven loci (MLVA7 is proposed to constitute a first line assay. The loci are compatible with moderate resolution equipment such as agarose gel electrophoresis and show a good congruence value with MLVA31. The associated MLVA and SNP data was imported together with published genotyping data by taking advantage of major enhancements to the MLVAbank software and web site. CONCLUSIONS: The present report provides a wide coverage of the genetic diversity of naturally occurring B. anthracis strains in France as can be revealed by MLVA. The data obtained suggests that once such coverage is achieved, it becomes possible to devise optimized first-line MLVA assays comprising a sufficiently low number of loci to be typed either in one multiplex PCR or on agarose gels. Such a selection of seven loci is proposed here, and future similar investigations in additional countries will indicate to which extend the same selection can be used worldwide as a common minimum set. It is hoped that this approach will contribute to an efficient and low-cost routine surveillance of important pathogens for biosecurity such as

  5. Failure of Sterne- and Pasteur-like strains of Bacillus anthracis to replicate and survive in the urban bluebottle blow fly Calliphora vicina under laboratory conditions.

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    Britta von Terzi

    Full Text Available This study aimed to elucidate the bacteriological events occurring within the gut of Calliphora vicina, selected as the European representative of blow flies held responsible for the spread of anthrax during epidemics in certain parts of the world. Green-fluorescent-protein-carrying derivatives of Bacillus anthracis were used. These lacked either one of the virulence plasmids pXO1 and pXO2 and were infected, or not infected, with a worm intestine phage (Wip4 known to influence the phenotype and survival of the pathogen. Blood meals were prepared for the flies by inoculation of sheep blood with germinated and, in case of pXO2+ strains, encapsulated cells of the four B. anthracis strains. After being fed for 4 h an initial 10 flies were externally disinfected with peracetic acid to ensure subsequent quantitation representing ingested B. anthracis only. Following neutralization, they were crushed in sterile saline. Over each of the ensuing 7 to 10 days, 10 flies were removed and processed the same way. In the absence of Wip4, strains showed steady declines to undetectable in the total B. anthracis counts, within 7-9 days. With the phage infected strains, the falls in viable counts were significantly more rapid than in their uninfected counterparts. Spores were detectable in flies for longer periods than vegetative bacteria. In line with the findings in both biting and non-biting flies of early workers our results indicate that B. anthracis does not multiply in the guts of blow flies and survival is limited to a matter of days.

  6. Molecular and insecticidal characterization of Vip3A protein producing Bacillus thuringiensis strains toxic against Helicoverpa armigera (Lepidoptera: Noctuidae).

    Science.gov (United States)

    Lone, Showkat Ahmad; Yadav, Radha; Malik, Abdul; Padaria, Jasdeep Chatrath

    2016-02-01

    Vegetative insecticidal proteins (Vip) represent the second generation of insecticidal proteins produced by Bacillus thuringiensis (Bt) during the vegetative growth stage of growth. Bt-based biopesticides are recognized as viable alternatives to chemical insecticides; the latter cause environmental pollution and lead to the emergence of pest resistance. To perform a systematic study of vip genes encoding toxic proteins, a total of 30 soil samples were collected from diverse locations of Kashmir valley, India, and characterized by molecular and analytical methods. Eighty-six colonies showing Bacillus-like morphology were selected. Scanning electron microscopy observations confirmed the presence of different crystal shapes, and PCR analysis of insecticidal genes revealed a predominance of the lepidopteran-specific vip3 (43.18%) gene followed by coleopteran-specific vip1 (22.72%) and vip2 (15.90%) genes in the isolates tested. Multi-alignment of the deduced amino acid sequences revealed that vip3 sequences were highly conserved, whereas vip1 and vip2 showed adequate differences in amino acid sequences compared with already reported sequences. Screening for toxicity against Helicoverpa armigera larvae was performed using partially purified soluble fractions containing Vip3A protein. The mortality levels observed ranged between 70% and 96.6% in the isolates. The LC50 values of 2 of the native isolates, JK37 and JK88, against H. armigera were found to be on par with that of Bt subsp. kurstaki HD1, suggesting that these isolates could be developed as effective biopesticides against H. armigera.

  7. Diagnostic real-time PCR assays for the detection of emetic Bacillus cereus strains in foods and recent food-borne outbreaks.

    Science.gov (United States)

    Fricker, Martina; Messelhäusser, Ute; Busch, Ulrich; Scherer, Siegfried; Ehling-Schulz, Monika

    2007-03-01

    Cereulide-producing Bacillus cereus can cause an emetic type of food-borne disease that mimics the symptoms provoked by Staphylococcus aureus. Based on the recently discovered genetic background for cereulide formation, a novel 5' nuclease (TaqMan) real-time PCR assay was developed to provide a rapid and sensitive method for the specific detection of emetic B. cereus in food. The TaqMan assay includes an internal amplification control and primers and a probe designed to target a highly specific part of the cereulide synthetase genes. Additionally, a specific SYBR green I assay was developed and extended to create a duplex SYBR green I assay for the one-step identification and discrimination of the two emesis-causing food pathogens B. cereus and S. aureus. The inclusivity and exclusivity of the assay were assessed using a panel of 100 strains, including 23 emetic B. cereus and 14 S. aureus strains. Different methods for DNA isolation from artificially contaminated foods were evaluated, and established real-time assays were used to analyze two recent emetic food poisonings in southern Germany. One of the food-borne outbreaks included 17 children visiting a day care center who vomited after consuming a reheated rice dish, collapsed, and were hospitalized; the other case concerned a single food-poisoning incident occurring after consumption of cauliflower. Within 2 h, the etiological agent of these food poisonings was identified as emetic B. cereus by using the real-time PCR assay.

  8. Preservation of Bacillus firmus Strain 37 and Optimization of Cyclodextrin Biosynthesis by Cells Immobilized on Loofa Sponge

    Directory of Open Access Journals (Sweden)

    Cristiane Moriwaki

    2012-08-01

    Full Text Available The preservation of Bacillus firmus strain 37 cells by lyophilization was evaluated and response surface methodology (RSM was used to optimize the β-cyclodextrin (β-CD production by cells immobilized on loofa sponge. Interactions were studied with the variables temperature, pH and dextrin concentration using a central composite design (CCD. Immobilization time influence on β-CD production was also investigated. B. firmus strain 37 cells remained viable after one year of storage, showing that the lyophilization is a suitable method for preservation of the microorganism. From the three-dimensional diagrams and contour plots, the best conditions for β-CD production were determined: temperature 60 °C, pH 8, and 18% dextrin. Considering that the amount of dextrin was high, a new assay was carried out, in which dextrin concentrations of 10, 15, and 18% were tested and the temperature of 60 °C and pH 8 were maintained. The results achieved showed very small differences and therefore, for economic reasons, the use of 10% dextrin is suggested. Increasing the immobilization time of cells immobilized on synthetic sponge the β-CD production decreased and did not change for cells immobilized on loofa sponge. The results of this research are important for microorganism preservation and essential in the optimization of the biosynthesis of CD.

  9. Mass spectrometric profiling of Bacillus cereus strains and quantitation of the emetic toxin cereulide by means of stable isotope dilution analysis and HEp-2 bioassay.

    Science.gov (United States)

    Stark, Timo; Marxen, Sandra; Rütschle, Andrea; Lücking, Genia; Scherer, Siegfried; Ehling-Schulz, Monika; Hofmann, Thomas

    2013-01-01

    A fast and robust high-throughput ultra-performance liquid chromatography/time-of-flight mass spectrometry (UPLC-TOF MS) profiling method was developed and successfully applied to discriminate a total of 78 Bacillus cereus strains into no/low, medium and high producers of the emetic toxin cereulide. The data obtained by UPLC-TOF MS profiling were confirmed by absolute quantitation of cereulide in selected samples by means of high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) and stable isotope dilution assay (SIDA). Interestingly, the B. cereus strains isolated from four vomit samples and five faeces samples from patients showing symptoms of intoxication were among the group of medium or high producers. Comparison of HEp-2 bioassay data with those determined by means of mass spectrometry showed differences, most likely because the HEp-2 bioassay is based on the toxic action of cereulide towards mitochondria of eukaryotic cells rather than on a direct measurement of the toxin. In conclusion, the UPLC-electrospray ionization (ESI)-TOF MS and the HPLC-ESI-MS/MS-SIDA analyses seem to be promising tools for the robust high-throughput analysis of cereulide in B. cereus cultures, foods and other biological samples.

  10. [Detection of toxigenic genes nheA, nheB and nheC in Bacillus cereus strains isolated from powdered milk samples in Costa Rica].

    Science.gov (United States)

    Rojas, Jonathan; Rodríguez-Rodríguez, Carlos E; Pérez, Cristian; Chaves, Carolina; Arias, María Laura

    2014-09-01

    Powdered milk is a frequently consumed product that does not need to be kept under cold conditions. Nevertheless, different microorganisms may contaminate it. Powdered milk is a highly consumed product by Costa Rican population, and Bacillus cereus is a potentially pathogenic bacteria associated to it, with the ability to develop toxins depending on the presence of the respective codifying genes. The aim of this study was to determine the presence of the toxigenic genes nheA, nheB and nheC from B. cereus strains, found in powdered milk sold at the Costa Rican national market. Five different lots of ten brands of powdered milk, distributed in the metropolitan area of San José, Costa Rica were analyzed. B cereus load was quantified using the Most Probable Number technique and identified using the Vitek system. The presence of the toxigenic genes was determined using the PCR technique. The isolation frequency of this bacteria in the powdered milk samples analyzed reached 50%, with populations ranging from 3 to > 100 MPN/g. Five out from nineteen strains were found positive for the three toxigenic genes, indicating contamination with potentially toxigenic B. cereus in powdered milk distributed in the national market, and an important risk for public. health.

  11. Inactivation kinetics of spores of Bacillus cereus strains treated by a peracetic acid-based disinfectant at different concentrations and temperatures.

    Science.gov (United States)

    Sudhaus, Nadine; Pina-Pérez, Maria Consuelo; Martínez, Antonio; Klein, Günter

    2012-05-01

    The purpose of this study was to assess the effect of a commercial peracetic acid-based disinfectant against spores of Bacillus cereus, to identify the most influential factor for the final number of microorganisms after different disinfection procedures, and to evaluate the nature of the inactivation kinetics. The spores of four different strains of B. cereus (DSM 318, 4312, 4313, and 4384) were treated with five different disinfectant concentrations (0.25%, 0.5%, 1.0%, 1.5%, and 2.0% [w/v]) at three different temperatures (10°C, 15°C, and 20°C) with or without protein load. A higher temperature and PES 15/23 concentration resulted in a higher inactivation. Inactivation of B. cereus strain 4312 was around 2 log₁₀ cycles at 10°C and around 7 log₁₀ at 20°C (conc=1% [w/v] PAA; t=60 min; without protein). The protein load at higher concentrations did not significantly reduce the efficacy of the disinfectant (p>0.05). This article indicates the applicability of the Weibull model to fit the B. cereus disinfectant survival curves. A Monte Carlo simulation was used to carry out a sensitivity analysis, which revealed the most influential factors affecting the final number of microorganisms after the disinfection process.

  12. Characterization and properties of biosurfactants produced by a newly isolated strain Bacillus methylotrophicus DCS1 and their applications in enhancing solubility of hydrocarbon.

    Science.gov (United States)

    Jemil, Nawel; Ben Ayed, Hanen; Hmidet, Noomen; Nasri, Moncef

    2016-11-01

    Six biosurfactant-producing bacteria were isolated from hydrocarbon contaminated soils in Sfax, Tunisia. Isolates were screened for biosurfactant production by different conventional methods including hemolytic activity, surface tension reduction, drop-collapsing and oil displacement tests. All these screening tests show that all the isolates behave differently. Among the isolated bacteria, DCS1 strain was selected for further studies based on its highest activities and it was identified as Bacillus methylotrophicus DCS1. This strain was found to be a potent producer of biosurfactant when cultivated in mineral-salts medium supplemented with diesel oil (2 %, v/v) as a sole carbon source. Physicochemical properties and stability of biosurfactants synthesized by B. methylotrophicus DCS1 were investigated. The produced biosurfactants DCS1, from Landy medium, possess high surface activity that could lower the surface tension of water to a value of 31 from 72 mN m(-1) and have a critical micelle concentration (CMC) of 100 mg L(-1). Compared with SDS and Tween 80, biosurfactants showed excellent emulsification activities against different hydrocarbon substrates and high solubilization efficiency towards diesel oil. Biosurfactants DCS1 showed good stability in a wide range of temperature, pH and salinity. These results suggested that biosurfactants produced by B. methylotrophicus DCS1 could be an alternative to chemically synthesized surfactants for use in bioremediation processes to enhance the solubility of hydrophobic compounds.

  13. Bacillus novalis sp. nov., Bacillus vireti sp. nov., Bacillus soli sp. nov., Bacillus bataviensis sp. nov. and Bacillus drentensis sp. nov., from the Drentse A grasslands.

    Science.gov (United States)

    Heyrman, Jeroen; Vanparys, Bram; Logan, Niall A; Balcaen, An; Rodríguez-Díaz, Marina; Felske, Andreas; De Vos, Paul

    2004-01-01

    A group of 42 isolates were isolated from the soil of several disused hay fields, in the Drentse A agricultural research area (The Netherlands), that were taken out of production at different times. The group represents hitherto-uncultured Bacillus lineages that have previously been found, by a non-cultural method, to be predominant in soil. The strains were subjected to a polyphasic taxonomic study, including (GTG)5-PCR, 16S rDNA sequence analysis, DNA-DNA hybridizations, DNA base-ratio determination, fatty acid analysis and morphological and biochemical characterization. By comparing the groupings obtained by (GTG)5-PCR and 16S rDNA sequence analysis, six clusters of similar strains could be recognized. A DNA-DNA relatedness study showed that these clusters represented five novel genospecies. Further analysis supported the proposal of five novel species in the genus Bacillus, namely Bacillus novalis sp. nov. (type strain IDA3307T=R-15439T=LMG 21837T=DSM 15603T), Bacillus vireti sp. nov. (type strain IDA3632T=R-15447T=LMG 21834T=DSM 15602T), Bacillus soli sp. nov. (type strain IDA0086T=R-16300T=LMG 21838T=DSM 15604T), Bacillus bataviensis sp. nov. (type strain IDA1115T=R-16315T=LMG 21833T=DSM 15601T) and Bacillus drentensis sp. nov. (type strain IDA1967T=R-16337T=LMG 21831T=DSM 15600T).

  14. 一株芽孢杆菌的分离鉴定及其益生潜质分析%Isolation and Identification of a Bacillus Subtilis Strain and its Potential Probiotic Analysis

    Institute of Scientific and Technical Information of China (English)

    徐海燕; 曹斌; 辛国芹; 曹银生; 武香玉; 陈静; 谢全喜; 谷巍

    2012-01-01

    A Bacillus strain BF3 ,which was isolated from the feces of healthy and adult broiler, was i-dentified as Bacillus subtilis by its cultural characteristics,morphological characteristics,physiological and biochemical properties and 16S rDNA sequence.Inhibition of the growth of enteric bacteria by Bacillus subtilis BF3 was studied.The results showed that the Bacillus subtilis BF3 could decrease the propagation of the Salmonella pullorum and Chicken Escherichia coli obviously in the course of incubation.The strength of the antagonism was influenced by the inoculation time of Bacillus and pathogenic.The Bacillus strain was inoculated firstly,the antagonism was the strongest.Bacillus and pathogenic bacteria were inoculated at the same time, the antagonism ranked the second.The Enteric bacterium was inoculated firstly,the antagonism was the weakest.And the Bacillus strain BF3 metabolites showed powerful ability to inhibit the growth of the Salmonella pullorum and Chicken Escherichia coli.%从健康肉鸡的粪便中分离出一株BF3芽孢杆菌,经培养特征、形态观察、生理生化试验以及16SrDNA序列分析相似度为99%,确定该菌株为枯草芽孢杆菌,对该株芽孢杆菌对肠道致病菌的拮抗作用进行研究.结果表明BF3菌株在培养过程中,可以抑制鸡白痢沙门氏菌和鸡大肠杆菌的生长.拮抗作用的强弱与芽孢杆菌和致病菌的接种时间有关.先接种芽孢杆菌后接种致病菌,拮抗作用最强;芽孢杆菌和致病菌同时接种次之;先接种致病菌后接种芽孢杆菌,拮抗作用最弱.并且BF3菌株代谢产物对鸡白痢沙门氏菌和鸡大肠杆菌具有非常明显的拮抗作用.

  15. Modular genetic architecture of the toxigenic plasmid pIS56-63 harboring cry1Ab21 in Bacillus thuringiensis subsp. thuringiensis strain IS5056.

    Science.gov (United States)

    Murawska, Emilia; Fiedoruk, Krzysztof; Swiecicka, Izabela

    2014-01-01

    Bacillus thuringiensis subsp. thuringiensis IS5056, a strain highly toxic to Trichoplusia ni larvae, produces the newly described Cry1Ab21 delta-endotoxin encoded by a gene located in the 63.8 kb pIS56-63 plasmid. In this report we present the structure and functional similarity of this plasmid to other B. thuringiensis large toxigenic plasmids with particular interest focused on its modular architecture. The 61 open reading frames (ORFs) of the plasmid made four functional modules: (i) M1-mic, the mobile insertion cassette harboring cry1Ab21; (ii) M2-tra, the putative conjugative element; (iii) M3-reg, regulation sequence; and (iv) M4-rep, the ori44 replicon. These modules display similarity to corresponding sequences in distinct B. thuringiensis plasmids, but, in general, not to plasmid of other Bacillus cereus sensu lato. The nucleotide sequence and organization of genes in pIS56-63 were highly similar (80-100%) to those in pHT73 of B. thuringiensis HT73, and in p03 of B. thuringiensis HD771, particularly within the M3-reg and M4-rep modules, and slightly less in M2-tra, the latter of which is composed of two segments exhibiting homology to sequences in pBMB28, pAH187_45, pCT83, and pIS56-85 or to pCT72, pBMB67, p04, and pIS56-68. The tetrapartite structure of the toxigenic pIS56-63 plasmid strongly suggests that its hybrid nature is a result of recombination of various genetic elements originating from different extrachromosomal and chromosomal sources in B. thuringiensis. The presence of cry1Ab21 in the mobile cassette suggests that its occurrence on pIS56-63 resulted from recombination and transposition events during the evolution of the plasmid.

  16. A Brazilian Bacillus thuringiensis strain highly active to sugarcane borer Diatraea saccharalis (Lepidoptera: Crambidae Uma Estirpe Brasileira de Bacillus thuringiensis com elevada atividade para a broca da cana-de-açúcar Diatraea saccharalis (Lepidoptera: Crambidae

    Directory of Open Access Journals (Sweden)

    Patrícia de Medeiros Gitahy

    2007-09-01

    Full Text Available The control of the major sugarcane pest, Diatraea saccharalis, is limited by the stem location of the caterpillar. As part of a long-term project towards the development of an alternative and efficient delivery system of Cry proteins to control the sugarcane borer, the current work describes the selection and characterization of a Brazilian B. thuringiensis strain with prominent activity towards D. saccharalis. Strain S76 was eleven-fold more active than the HD-1 Lepidoptera-standard strain, as estimated by the LC50 of 13.06 µg/L and 143.88 µg/L, respectively. We observed bipiramidal and cuboidal crystals similar to those found in other B. thuringiensis strains with entomopathogenic activity against Lepidoptera and Diptera. In addition, smaller and spherical crystalline inclusions were also observed. The plasmid profile of strain S76 is similar to that of HD-1. PCR amplifications of S76 DNA using cry specific primers confirmed the presence of cry1Aa,cry1Ab,cry1Ac,cry2Aa1, and cry2Ab2, but not cry1Ad, cry2Ac and cry9 type genes. No differences that could explain the superior activity of S76 when compared to HD-1, the Lepidoptera standard strain, were observed. Nevertheless, its higher entomopathogenic activity has pointed this strain S76 as a potential source of cry genes to control sugarcane borer, an important pest that affects sugarcane, a crop that occupies a planted area of about 6 million ha in Brazil.Diatraea saccharalis é o inseto-praga que provoca os maiores danos a cultura da cana-de-açúcar, e seu controle é limitado pela localização do ataque no interior do colmo das plantas. Como parte de um projeto a longo prazo com o objetivo de desenvolver uma alternativa eficiente para o controle da broca da cana utilizando as proteínas Cry de Bacillus thuringiensis, o presente trabalho descreve a seleção e caracterização de uma estirpe desta bactéria com atividade larvicida para D. saccharalis. A estirpe brasileira S76, foi

  17. Cyclodextrin glycosyltransferase production by new Bacillus sp. strains isolated from brazilian soil Produção de ciclodextrina glicosiltransferase por novas cepas de Bacillus sp. isoladas de solo brasileiro

    Directory of Open Access Journals (Sweden)

    Vivian Menocci

    2008-12-01

    Full Text Available Three strains of Bacillus sp. (BACRP, BACNC-1 and BACAR were isolated from soil adhered to cassava husk. CGTase specific activity for the three isolated strains was higher when cultivated at 40ºC. Potato starch, cassava starch, maltodextrin and glucose were used as carbon source and growth temperatures varied from 25 to 55ºC. The three isolates presented higher CGTase specific activity when cultivated with potato starch at 40ºC. Isolated BACRP and BACAR presented specific activity of 4.0x10-3 and 2.2x10-3 U/mg prot at pH 7.0, respectively, when cultivated in mediums added with NaCl 2%; at pH 10,0 their activities were of 3.4x10-3 and 3.0x10-3 U/mg prot, respectively, in the same concentration of NaCl. On the other hand, the isolated BACNC-1 presented activity specific of 2.4x10-3 U/mg prot when cultivated at pH 7.0 added of NaCl 1%, and at pH 10.0 the specific activity was of 3.4x10-3 U/mg prot without NaCl addition. This work also showed the presence of cyclodextrins formed during fermentation process and that precipitation with acetone or lyophilization followed by dialysis was efficient at removing CDs (cyclodextrins, thus, eliminating interference in the activity assays. The enzyme produced by the BACAR strain was partially purified and β-CD was liberated as a reaction product.Três linhagens de Bacillus sp (BACRP, BACNC- 1 e BACAR foram isoladas a partir de solo aderido em casca de mandioca. Foram utilizados amido de batata, amido de mandioca, maltodextrina e glicose como fonte de carbono, e temperaturas de crescimento de 25-55ºC, sendo que os três isolados apresentaram maior atividade específica de CGTase quando cultivados com amido de batata a 40ºC. Em pH 7,0 os isolados BACRP e BACAR apresentaram atividade específica de 4,0x 10-3 e 2,2x10-3 U/mg prot, respectivamente, quando cultivados em meios acrescidos de 2% de NaCl; em pH 10,0 suas atividades foram de 3,4x10-3 e 3,0x10-3 U/mg prot na mesma concentração de NaCl. Por outro

  18. 中国吐鲁番两株产木聚糖酶的极端耐碱Bacillus halodurans的分类鉴定%Isolation and identification of two xylanase-producing extremely alkali-tolerant strains of Bacillus halodurans from Turpan in China

    Institute of Scientific and Technical Information of China (English)

    易霞; 谢周杰; 邓爱华; 王宁; 艾尔肯·热合曼

    2006-01-01

    通过生理生化实验、16S rDNA 序列分析和同源性杂交,将分离到的XJU-1和XJU-80菌种进行了分类鉴定.XJU-1和XJU-80具有较宽的pH生长范围(分别是pH4.5~12.6和pH3.8~12.6),其G+C mol%含量分别是40.5mol%和42.2mol%.16S rDNA 序列分析和DNA-DNA同源杂交结果表明,XJU-1和XJU-80与Bacillus halodurans C-125和Bacillus halodurans DSM497T具有较高的同源性(99%);两者之间也具有85%的相关性,但其与Bacillus halodurans C-125和Bacillus halodurans DSM497T分别具有81.3%和71.5%的相关性.基于以上结果,将两株分离菌株分类为Bacillus halodurans的两个新品系.%Bacillus halodurans XJU-1 and XJU-80 were characterized in terms of physiological and biochemical characteristics, and 16S rDNA sequence homology and DNA-DNA hybridization analysis. The two isolates can grow in nutrient broth at a broad range of pH values from 4.5 to 12.6 for XJU-1 and from 3.8 to 12.8 for XJU-80, respectively. And the optimum temperature of growth were around 39℃and 42℃, respectively. Phylogenetic analysis of the two strains based on comparison of 16S rRNA sequence revealed that they are closely related to Bacillus halodurans C-125 and DSM497Twith 99% identity. DNA-DNA hybridization showed that the highest levels of DNA-DNA relatedness were found between the two strains (85%) and the B. Halodurans type strains (81.3% and 71.5%), respectively. Moreover, the G+C content of the genomic DNA was 40.5 mol% for XJU-1 and 42.2 mol% for XJU-80.Our results demonstrate that strains XJU-1 and XJU-80 should be classified as two new members of the species B. Halodurans.

  19. Bacillus coagulans

    Science.gov (United States)

    Bacillus coagulans is a type of bacteria. It is used similarly to lactobacillus and other probiotics as "beneficial" bacteria. People take Bacillus coagulans for diarrhea, including infectious types such as ...

  20. Seleção e caracterização de estirpes de Bacillus thuringiensis efetivas no controle da traça-das-crucíferas Plutella xylostella Selection and characterization of Bacillus thuringiensis strains effective to control the diamondback moth Plutella xylostella

    Directory of Open Access Journals (Sweden)

    Patrícia Teles Medeiros

    2005-11-01

    Full Text Available O objetivo deste trabalho foi selecionar e caracterizar, no Banco de Germoplasma de Bacillus spp., da Empresa Brasileira de Pesquisa Agropecuária, as estirpes de Bacillus thuringiensis mais tóxicas à Plutella xylostella, por métodos morfológicos, bioquímicos e moleculares. Das 203 estirpes testadas, sete causaram 100% de mortalidade e foram semelhantes à estirpe padrão utilizada, B. thuringiensis subsp. kurstaki. Elas apresentaram proteínas de 130 kDa e 65 kDa, presença de genes cry1 e cry2 e cristais bipiramidais, cubóides e redondos. As estirpes selecionadas oferecem novas perspectivas de controle de P. xylostella.The aim of this work was to select and characterize the most toxic Bacillus thuringiensis strains, from the Germplasm Bank of Bacillus spp. of Empresa Brasileira de Pesquisa Agropecuária, against Plutella xylostella. Strains were characterized by morphological, biochemical and molecular methods. It was observed that seven out of the 203 strains tested showed high toxicity compared to the standard used B. thuringiensis subsp. kurstaki (HD-1, which showed 100% mortality. Selected strains showed features described for lepidoptera regarding the protein of 130 kDa and 65 kDa; profile and features were obtained through the PCR reactions, making possible to identify the presence of cry1 and cry2 genes. Moreover, the scanning electron microscopy showed the bipiramydal, cubed and round crystal forms. The selected strains offer new perspectives to control P. xylostella.