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Biodegradation of agribiomass especially wheat straw to biohydrogen and biomethane is an encouraging approach to the current waste management problem. To do so, the biomass must first be pretreated to break down lignin thereby increasing accessibility of the substrate to fermentative organisms. In the current study, out of 20 isolates from the granular sludge of full-scale anaerobic digester, four ligninolytic Bacillus sp. strains were selected based on their lignin and Azure B degradation. Further, among the four isolates, Brevibacillus agri AN-3 exhibited the highest of 88.4 and 78.1% decrease in COD of lignin and Azure B respectively. These strains were also found to secrete optimum yields of lignin peroxidase (LiP) at pH 3, laccase (Lac) at pH 5, and xylanase and cellulase enzymes at pH 7. The strains demonstrated maximum activity of Lip and Lac at 50 °C and xylanase and cellulase at 60 °C after 72-h growth. Among the four strains, Brevibacillus agri AN-3 showed hydrogen (H2) yield of 1.34 and 2.9 mol-H2/mol from xylose and cellulose respectively. In two-phase wheat straw batch fermentation, Brevibacillus agri AN-3 produced 88.3 and 283.7 mL/gVS cumulative H2 and methane (CH4) respectively. Biotreatment with ligninolytic Bacillus sp. strains perceived that 261.4% more methane yield could be obtained from the wheat straw than using the untreated wheat straw in batch fermentation. This is the first study establishing not only the hydrogen potential of ligninolytic Bacillus sp. strains but also indicates a vital role of these species in developing standard inoculum and a biocatalyst for processing agribiomass.ᅟ [ABSTRACT FROM AUTHOR]

Abstract The detergent-compatible alkaline protease was produced from the bacterial strain Bacillus sp. APP-07 isolated from Laundromat soil of Solapur, Maharashtra, India. The culture was grown in 1000 ml capacity baffled flask with a working volume of 100 ml and incubated at 55 °C for 33 h on a rotary shaker. After incubation, alkaline protease was partially purified by the sequential method of acetone precipitation followed by nominal molecular weight limit (NMWL) cut-off ultrafiltration using 50 K and 10 K filters. Finally, Sephadex G-100 gel filtration chromatographic purification was performed to obtain 3.12 fold purified alkaline protease enzyme with a 66.67% final yield. The purified enzyme showed 31907.269 units (U) of enzyme activity containing 8741.718 U/mg of specific enzyme activity. The molecular weight of the enzyme was confirmed about 33.0 kDa (kDa) by the SDS-PAGE analysis. The purified enzyme was stable at higher pH and temperature range, with an optimum pH 10.5 and temperature 55 °C. The enzyme showed excellent stability and compatibility in various detergents, surfactants, bleach, and oxidizing agents. The enzyme activity enhanced in the presence of Ca2+, Cu2+, and surfactants, whereas; the phenylmethylsulphonyl fluoride (PMSF) and Diisopropyl fluorophosphate (DFP) completely inhibit the enzymatic activity, which pointed out that the enzyme affiliated to serine-centered metalloproteases family. In conclusion, the remarkable tolerance and stability of the enzyme explored the promising candidature for the several potential applications in the laundry detergents. The sustainability of the enzyme might serve several possible applications in the laundry detergents, leather industries, and other harsh industrial processes. [ABSTRACT FROM AUTHOR]

Abstract Inulinase are industrial food enzymes which have gained much attention in recent scenario. In this study, Inulinase producing eight bacterial colonies were isolated and screened from three different plant root tubers soil sample. Among 8 inulinase producing colonies, the higher yielding colony was selected with 25.10 U/mL for further studies. The best inulinase producing colony was identified by partial 16S rRNA gene sequence as Bacillus sp. The crude inulinase was purified by using ammonium sulphate precipitation, dialysis and ion exchange chromatography on DEAE – sephacel and obtained 1.9 purification fold with total activity 293 U. The purified enzyme was subjected to characterization studies and it was found to be stable at 30–60 °C and optimum temperature was at 55 °C. The enzyme was stable at pH 3.0–7.0 and optimum pH was at 6.5. The K m and V max value for inulinase was found to be 0.117 mg/mL and 4.45 μmol min mg−1 respectively, demonstrate its greater affinity. Hence, this enzyme can be widely used for the production of fructose, and fructooligosaccharides, which are important ingredients in food and pharmaceutical industry. [ABSTRACT FROM AUTHOR]

Abstract Fungal and bacterial pathogens infect a diverse range of hosts including various plant and animal species. Fungal and bacterial diseases, especially of plants and aquatic animals, such as fish, lead to significant damage to crops and aquaculture, respectively, worldwide. The present study was conducted to isolate and characterize potent Bacillus strains with significant antagonistic activity against the major plant and fish pathogenic fungi and bacteria. We randomly collected 22 isolates of Bacillus from the soil, rhizosphere, and sediment from different parts of Bangladesh. Initial characterization, based on in vitro antagonistic activity on the culture plate, resulted in the selection of four gram-positive Bacillus sp. isolates. Among these, the isolate BC01, obtained from soil demonstrated the highest broad-spectrum anti-bacterial and anti-fungal activities. We confirmed the genus of BC01 to be Bacillus by morphological and biochemical tests as well as using molecular data analysis tools, including the study of 16s rDNA, phylogenetic relationship, and evolutionary divergence scores. The isolate significantly inhibited the mycelial growth of the plant pathogen, Penicillium digitatum and fish pathogen, Aphanomyces invadans in vitro. The anti-bacterial effect of the isolate was also evaluated against Pseudomonas spp. and Xanthomonas spp., the two deadliest plant pathogens, and Aeromonas veronii , Pseudomonas fluorescens , and Streptococcus iniae , three major fish pathogens that are primarily responsible for global aquaculture loss. The results of the present study could pave the way for developing potent drugs to combat microbial infection of plants and fish. [ABSTRACT FROM AUTHOR]

Bacillus sp. SJ-10 (KCCM 90078, JCM 15709) is a halotolerant bacterium isolated from a traditional Korean food, i.e., salt-fermented fish (jeotgal). The bacterium can survive and engage in metabolism at high salt concentrations. Here, we reported complete genome sequence of Bacillus sp. SJ-10, which has a single circular chromosome of 4,041,649 base pairs with a guanine-cytosine content of 46.39%. Bacillus sp. SJ-10 encodes a subunit of poly-γ-glutamic acid (γ-PGA) with a molecular weight of approximately 400 kDa, which contains four γ-PGA synthases (pgsB, pgsC, pgsAA and pgsE) and one γ-PGA-releasing gene (pgsS). This bacterium also able to produce salt-stable enzymes such as protease, β-glucosidase, and β-1,3-1,4-glucanase. This affords significant insights into strategies employed by halotolerant bacteria to survive at high salt concentrations. The sequence contains information on secondary metabolites biosynthetic gene cluster, and most importantly enzymes produced by the bacterium may be valuable with respect to food, beverage, detergent, animal feed, and certain commercial contexts. [ABSTRACT FROM AUTHOR]

βγ-crystallin has emerged as a superfamily of structurally homologous proteins with representatives across all domains of life. A major portion of this superfamily is constituted by microbial members. This superfamily has also been recognized as a novel group of Ca2+-binding proteins with a large diversity and variable properties in Ca2+ binding and stability. We have recently described a new phosphatidylinositol phospholipase C from Lysinibacillus sphaericus (LS-PIPLC) which was shown to efficiently remove phosphatidylinositol from crude vegetable oil. Here, the role of the C-terminal βγ-crystallin domain of LS-PIPLC was analyzed in the context of the whole protein. A truncated protein in which the C-terminal βγ-crystallin domain was deleted (LS-PIPLCΔCRY) is catalytically as efficient as the full-length protein (LS-PIPLC). However, the thermal and chemical stability of LS-PIPLCΔCRY are highly affected, demonstrating a stabilizing role for this domain. It is also shown that the presence of Ca2+ increases the thermal and chemical stability of the protein both in aqueous media and in oil, making LS-PIPLC an excellent candidate for use in industrial soybean oil degumming. [ABSTRACT FROM AUTHOR]

Antimicrobial peptides (AMPs) constitute a major arm of defense in mosquitoes against microbes. The purpose of this study was to determine which of the peptides are produced in the mosquito hemolymph after bacterial treatment. Culex pipiens (Diptera: Culicidae) larvae, collected from drainage canal in Suez Governorate, Egypt, were treated with LC80 of Bacillus sphaericus strain 2362 (0.035 ppm). The hemolymph of bacteria-treated Cx. pipiens mosquitoes and non-treated mosquitoes as control were extracted and undergo electrophoresis, using a Bio-Rad Mini-protean II cell. In Cx. pipiens mosquitoes emerging from bacteria-treated larvae, a band of protein was detected at 165 kDa. This protein was more likely to be Thioester-containing protein 1 (TEP1). Understanding mosquito resistance mechanism to bacterial control is crucial to build up programs to overcome such resistance. [ABSTRACT FROM AUTHOR]

The ultrastructure alterations were described in the midgut of Culex pipiens (Diptera: Culicidae) larvae following Bacillus sphaericus treatment. Two hours post-treatment, the epithelial layer began to have large amount of vacuoles. Explosion in the basement membrane took place 6 h post-treatment. Ten hours post-treatment, the bacterium began to invade the microvilli. At 14 h post-bacterial treatment, epithelial cells were destroyed, microvilli were damaged, and bacteria were present in lumen in large amount. The present study provided the evidence on the main aberrations induced in midgut larvae of Cx. pipiens as a result of ingesting B. sphaericus. [ABSTRACT FROM AUTHOR]

Mosquitoes are the most critical group of insects in the context of public health, since they transmit key parasites and pathogens, causing millions of deaths annually. Insecticides from natural products may boost the effectiveness of vector control programs. In this study, we tested silver nanoparticles (AgNPs) fabricated using the leaf extract of the orchid Zeuxine gracilis as reducing agent, and the microbial pesticide Bacillus sphaericus, against the mosquitoes Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus. The synthesis of AgNP was confirmed analyzing the excitation of surface Plasmon resonance using ultraviolet–visible (UV–Vis) spectrophotometry. SEM and TEM showed the irregular shapes of AgNPs. EDX spectroscopy, FTIR spectroscopy, X-ray diffraction and dynamic light scattering analysis were carried out. AgNPs were highly effective against the larvae of An. stephensi (LC50 = 8.48 µg/mL), Ae. aegypti (LC50 = 10.39 µg/mL) and Cx. quinquefasciatus (LC50 = 13.21 µg/mL), respectively. Combined treatments testing B. sphaericus with AgNPs were also effective against An. stephensi (LC50 = 12.32 µg/mL), Ae. aegypti (LC50 = 14.78 µg/mL) and Cx. quinquefasciatus (LC50 = 19.19 µg/mL). Overall, this study suggests that the orchid-synthesized AgNPs can be a rapid, environmentally safer bio-pesticide to be used in synergy with B. sphaericus to control mosquito vectors. [ABSTRACT FROM AUTHOR]

The suitability of using a spore-forming ureolytic strain, Bacillus sphaericus, was evaluated for self-healing of concrete cracks. The main focus was on alkaline tolerance, calcium tolerance, oxygen dependence, and low-temperature adaptability. Experimental results show that B. sphaericus had a good tolerance. It can grow and germinate in a broad range of alkaline pH. The optimal pH range is 7 ∼ 9. High alkaline conditions (pH 10 ∼ 11) slow down but not stop the growth and germination. Oxygen was strictly needed during bacterial growth and germination, but not an essential factor during bacterial urea decomposition. B. sphaericus also had a good Ca tolerance, especially at a high bacterial concentration of 10 cells/mL; no significant influence was observed on bacterial ureolytic activity of the presence of 0.9M Ca. Furthermore, at a low temperature (10 °C), bacterial spores germinated and revived ureolytic activity with some retardation. However, this retardation can be counteracted by using a higher bacterial concentration and by supplementing yeast extract. It can be concluded that B. sphaericus is a suitable bacterium for application in bacteria-based self-healing concrete. [ABSTRACT FROM AUTHOR]

Diaphorases are flavin-containing enzymes with potential applications in biotransfomation reactions, biosensor design and in vitro diagnostic tests. In this communication, we describe recombinant expression, characterization and application of a lipoamide dehydrogenase (DLD) with diaphorase activity from a strain of Bacillus sphaericus. The DLD gene consisting of 1413 bp encoding a protein of 470 amino acids was expressed in Escherichia coli BL21 (DE3) and the recombinant enzyme was characterized. B. sphaericus DLD catalyzed the reduction of NAD by dihydrolipoamide and exhibited NADH-dependent diaphorase activity. The molecular weight of purified enzyme was about 50 kDa, and determined to be a monomeric protein. Diaphorase was active and stable from pH 7.0 to 9.0 with an optimal activity at pH 8.5. It showed its maximal activity at temperature of 30 °C and was almost stable at temperatures between 25 and 30 °C. Different metal ions and inhibitors showed no influence on the activity of target enzyme. The K and V values for NADH were estimated to be 0.33 mM and 200.0 U/ml, respectively. Moreover, recombinant B. sphaericus diaphorase exhibited considerable potential to be used as a component of diagnostic tests for the quantification of metabolites. In conclusion, considering the properties of diaphorase from B. sphaericus PAD-91, it can have potential application as a diagnostic enzyme. [ABSTRACT FROM AUTHOR]

Mosquitoes are important vectors of many infectious diseases. Bacillus sphaericus (Bs) is an ideal larvicide and has attracted more and more attention, recently. However, the fundamental research of its application is very limited, especially on the subsequent impact of Bs exposure on mosquito's fecundity and resistance emergence. Through bioassay, LC50 and LC95 of Bs in killing Anopheles dirus larvae were determined as 9.793 ± 1.878 IU/L and 62.4 ± 6.438 IU/L at 48 h posttreatment, 7.128 ± 0.913 IU/L and 34.385 ± 12.547 IU/L at 72 h post treatment, respectively. After being treated with a sub-lethal dose of Bs, gravidity, oviposition, hatch, pupation, and eclosion of the surviving mosquitoes were counted and analyzed to elucidate the subsequent effects of Bs exposure on the reproductive capacity of A. dirus. The result interestingly showed that the exposure of Bs significantly reduced the oviposition ability of the surviving A. dirus, without effect on egg formation/gravidity, hatch, pupation, and eclosion. The surviving mosquitoes were also maintained routinely for generations to test the sustained effect of Bs exposure on the fecundity of the offsprings. After conventional breeding for generations, the capacity of egg laying totally recovered. To explore the rules of resistance development, bioassays were performed after treatment twice with a sub-lethal dose of Bs on two continuous generations of A. dirus larvae. The killing efficacies between the Bs treated group and control group were compared. The results showed that LC50 and LC95 increased by 4.35- and 7.37-folds after treatment with the sub-lethal dose of Bs on two consecutive generations, respectively. The results indicated that A. dirus was sensitive to Bs, which could reduce oviposition of the surviving A. dirus. The subsequent effect might help to further decrease the mosquito population. However, a sub-lethal dose of Bs exposure could easily cause resistance development. Our study provides a dose standard and reference for the rational use of Bs, which will be helpful for mosquito control. [ABSTRACT FROM AUTHOR]

Biological removal of sulfide, nitrate, and phenol at loading rates of 600 g S/(m day), 900 g N/(m day), and 450 g C/(m day), respectively, from synthetic wastewaters was achieved in an expanded granular sludge bed (EGSB) reactor, whose rates are much higher than literature works and are considered feasible for handling high-strength petrochemical wastewaters without dilution. Effects of C/S ratio (2-2.5:1) on EGSB performance were noted insignificantly. The strains Bacillus sp., Thauera sp., and Pseudomonas sp. were the heterotrophic denitrifiers and the strains Thiobacillus sp., Azoarcus sp., and Sulfurovum sp. were the autotrophic denitrifiers in the EGSB granules. The EGSB reactor experienced biological breakdown at loadings higher than 1200 g S/(m day), 1800 g N/(m day), and 900 g C/(m day) by the following mechanism: high sulfide first inhibits heterotrophic denitrifies ( Bacillus sp. and Pseudomonas sp.), thereby accumulating nitrite in the system; then, the accumulated nitrite inhibits autotrophic denitrifiers ( Thiobacillus sp., Azoarcus sp., and Sulfurovum sp.) to complete breakdown of the system. [ABSTRACT FROM AUTHOR]

The beneficial effect of microbially induced carbonate precipitation on building materials has been gradually disclosed in the last decade. After the first applications of on historical stones, promising results were obtained with the respect of improved durability. An extensive study then followed on the application of this environmentally friendly and compatible material on a currently widely used construction material, concrete. This review is focused on the discussion of the impact of the two main applications, bacterial surface treatment and bacteria based crack repair, on concrete durability. Special attention was paid to the choice of suitable bacteria and the metabolic pathway aiming at their functionality in concrete environment. Interactions between bacterial cells and cementitious matrix were also elaborated. Furthermore, recommendations to improve the effectiveness of bacterial treatment are provided. Limitations of current studies, updated applications and future application perspectives are shortly outlined. [ABSTRACT FROM AUTHOR]

Traditional TiO has been used as an antimicrobial additive to paints, but more recently the use of TiO nanoparticles (NPs) has been proposed as an alternative because of its ability to induce oxidative damage to the cell membrane of bacteria. This study focused on how photoinactivation of TiO NPs by fluorescent and halogen lights (400-700 nm) influenced survival of Bacillus sphaericus (Gram-positive bacterium) and Klebsiella pneumoniae (Gram-negative bacterium) isolated from spoiled paints. The loss of viability of the test organisms in the presence of TiO NPs determined by culturable (plate) count technique indicated a decrease in viable bacteria that was predominant after 24-h exposure. The TiO NPs showed higher antibacterial performance under fluorescent light than halogen light with increasing irradiation time and confirms the photokilling effect of TiO NPs. TiO NPs were also bactericidal under dark conditions, suggesting potential antibacterial applications in the paint industry. [ABSTRACT FROM AUTHOR]

This paper discusses the biosorption of Ni(II) ions from aqueous solutions by Bacillus sphaericus biomass. The biosorption process was affected by the solution pH, biomass concentration, contact time, temperature and initial Ni(II) concentration respectively. The sorption kinetics and equilibrium data were described well with the pseudo-second order kinetic model and the Freundlich isotherm model. The maximum monolayer biosorption capacity value of Bacillus sphaericus biomass for Ni(II) ions was calculated at 55.55 mg/g using the Langmuir isotherm model. The thermodynamic study shows the Ni(II) biosorption was spontaneous and exothermic in nature. The change in heat of sorption (ΔHo) and the isosteric heat of sorption (ΔHr) values indicate the physical sorption as the predominant mechanism for Ni(II) biosorption. The Ni(II) ions were recovered effectively from Bacillus sphaericus biomass using 0.1 M HNO3 and can be recycled. FTIR results showed that carboxylic and amine groups of Bacillus sphaericus cells were responsible for Ni(II) binding. [ABSTRACT FROM AUTHOR]

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

Ecotoxicity tests are key to predict environmental hazards resulting from chemical and biological pesticides in non-target species. In order to assess the effects of microbial pesticides it is important to determine if they cause infection in test organisms. At present the microbial elimination rate or clearance is not included in ecotoxicological regulatory protocols. This study evaluated the elimination of Bacillus thuringiensis and Bacillus sphaericus from fish and snails, after 30 days' exposure to commercial formulations of such entomopathogens. Data obtained showed that in clean water the tendency to eliminate microbial agents from the body of the exposed organisms is gradual over time but after 7 days the fish and snails were free of the two tested Bacillus spp. [ABSTRACT FROM AUTHOR]

Bacterial insecticides like, Bacillus sphaericus and Bacillus thuringiensis serovar israelensis, have been used for the control of nuisance and vector mosquitoes for more than two decades. For many years, it was assumed that the use of microbial larvicides based on B. sphaericus would not lead to resistance in mosquitoes. However, recent reports have shown that B. sphaericus toxins are not free from this problem. Therefore, the resistance of mosquito populations to be will seriously threaten the sustainability of current mosquito control programme using these microbial insecticides. In the present study, we have characterised a novel protein responsible for resistance development in the filariasis vector of Culex quinquefasciatus. Laboratory selection experiments with B. sphaericus against the larvae were carried out up to 17 generations, and the occurrence of resistance was reported (resistance ratio (RR) at lethal concentration (LC) and LC = 1,987 and 2,051 folds, respectively). The protein profiles of B. sphaericus-resistant and susceptible population have confirmed with the expression of a new polypeptide (80 kDa) in the resistant strain only. Sequence result revealed that the newly expressed protein was 'hexamerin', and this factor might conceivably be responsible for the inheritance of resistance. This study is therefore valuable for comprehending the underlining factor and management of B. sphaericus resistance problem in mosquito population. [ABSTRACT FROM AUTHOR]

The mature surface layer (S-layer) protein SlpC of mosquitocidal Bacillus sphaericus C3-41 comprises amino acids 31-1,176 and could recrystallize in vitro. The N-terminal SLH domain is responsible for binding function. Deletion of this part, S-layer proteins could not bind to the cell wall sacculi. To investigate the self-assembly ability of SlpC from B. sphaericus, nine truncations were constructed and their self-assembly properties were compared with the recombinant mature S-layer protein rSlpC. The results showed that rSbsC and truncations rSlpC, rSlpC, rSlpC, and rSlpC could assemble into multilayer cylinder structures, while N-terminal truncations rSlpC, rSlpC, and rSlpC mainly showed monolayer cylinders in recombinant Escherichia coli BL21 (DE3) cells. Growth phase analysis of the self-assembly process revealed that rSlpC mainly formed monolayer cylinders in the early stage (0.5 and 1 h induction of expression), but few double-layer or multilayer cylinders were also found with the cells growing, while rSlpC could formed multilayer cylinders in all the growth stage in the E. coli cells. It is concluded that the deletion of the C-terminal 126 aa or the N-terminal 497 aa did not interfere with the self-assembly process, the fragment (amino acids 278 to 337) is essential for the multilayer cylinder formation in E. coli BL21 (DE3) cells in the early stage and the fragment (amino acids 338 to 497) is related to monolayer cylinder formation. The information is important for further studies on the assembly mechanism of S-layer proteins and forms a basis for further studies concerning surface display and nanobiotechnology. [ABSTRACT FROM AUTHOR]

A Bacillus sp. isolated from sediments of distillery unit was found to overproduce laccase when cultured in a synthetic media containing 1mM CuSO and 10% distillery spent wash as inducers along with 1% dextrose (w/v) and 0.1% tryptone (w/v) as additional carbon and nitrogen sources. The extracellular purified enzyme was highly thermostable with a calculated half-life of 23 min at 75°C. The optimal pH and temperature of the Bacillus sp. laccase were recorded to be 3.0 and 35°C, respectively. Sodium azide and solvents like methanol and acetonitrile completely inhibited enzyme activity. The average molecular weight of the purified enzyme as determined by SDS-PAGE and zymogam studies was around 70 kDa. Kinetic parameters were detected by using 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulfonate) ( ABTS) as substrate. At high ABTS concentrations (> 6 mM) a substrate inhibition phenomenon appeared and K (0.60 mM), V (983.00 U/min) values were determined. The polypeptide sequences showed significant similarity with Cudependent oxidoreductases through MALDI-TOF MS analysis. In addition, the crude Bacillus sp. laccase showed enormous potential for decolorization of various recalcitrant dyes. The apparent high stability of this enzyme makes it a good candidate for its possible application in biotechnology. [ABSTRACT FROM AUTHOR]

The efficacy of copepod Mesocyclops aspericornis (Daday) combined with the larvicide Bacillus sphaericus (Bs) and a plant extract of Plectranthus amboinicus leaf abstract (PALE), used jointly and singly, was studied against Aedes aegypti in the laboratory. P. amboinicus leaf extract of 20, 40, 60, 80 and 100 ppm caused significant mortality against Ae. aegypti larvae. The LC and LC values for I to IV instars larvae and pupae were 26.12, 35.36, 45.76, 52.32 and 63.82 ppm, respectively. The LC values for I to IV instars larvae and pupae were 82.53, 92.65, 108.06, 119.47 and 131.71 ppm, respectively. Under laboratory conditions, copepods treatment produced 7.9% predatory efficiency against 1st instar larvae of Ae. aegypti, at a copepod:larvae ratio of 1:10. When copepod treatment was combined with PALE this was increased to 8.7. The treatment of copepods combined with Bs and PALE yielded a better and more sustainable result (9.6%) than the agents used individually. This predation efficiency may be caused by detrimental effects of the P. amboinicus active principle compound (carvacrol) on the mosquito larvae. Our results suggest that the combined application of microbial insecticide (Bs), copepods and P. amboinicus leaf extract may be used to control Aedes populations. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Arthrobacter sp. Sphe3 and Bacillus sphaericus cells were used for Cu(II) biosorption. The effect of contact time, biosorbent dose, equilibrium pH, temperature and the presence of other ions on the efficiency of the process were extensively studied. Optimum pH value and biomass concentration were determined at 5.0 and 1.0 g/l, whereas contact time was found to be 5 and 10 min for Arthrobacter sp. Sphe3 and Bacillus sphaericus biomass, respectively. Equilibrium data fitted very well to Freundlich model ( R = 0.996, n = 2.325, K = 8.141) using Arthrobacter sp. Sphe3. In the case of B. sphaericus, a Langmuir adsorption model [ R = 0.996, Q = 51.54 mg-Cu(II)/g] showed to better describe the results. Potentiometric titration and Fourier transform infrared (FTIR) spectroscopy showed that amine, carboxyl and phosphate groups participate in Cu(II)-binding. The calculated thermodynamic parameters indicated the spontaneous and feasible nature of Cu(II) biosorption on both biosorbents. Selectivity of Cu(II) biosorption was examined in binary and multi-ions systems with various anions and cations which are commonly found in municipal and industrial wastewater. A specificity towards Cu(II) was observed in binary mixtures with Cl, CO, NO, SO, PO, Mg and Ca, and As(V) with the maximum uptake capacity remaining constant even at high competitive ion's concentrations of 200 mg/l. Desorption studies showed that Cu(II) could be completely desorbed from Cu(II)-loaded Arthrobacter strain Sphe3 and B. sphaericus biomass using 1.0 and 0.8 M HCl, respectively, and both bacterial species could be effectively reused up to five cycles, making their application in wastewater detoxification more attractive. [ABSTRACT FROM AUTHOR]

Bacillus sphaericus produces a two-chain binary toxin composed of BinA (42 kDa) and BinB (51 kDa), which are deposited as parasporal crystals during sporulation. The toxin is highly active against Culex larvae and Aedes and Anopheles mosquitoes, which are the principal vectors for the transmission of malaria, yellow fever, encephalitis, and dengue. The use of B. sphaericus and Bacillus thuringiensis in mosquito control programs is limited by their sedimentation in still water. In this study, the binA and binB genes were cloned and the recombinant BinAB protein was expressed in three strains of Escherichia coli. These recombinant strains were used in a toxicity assay against Culex quinquefasciatus larvae. The highest expression level was achieved when both proteins were expressed in a single operon construct. The BinAB protein expressed in the E. coli Arctic strain showed higher larvicidal activity than either of the recombinant proteins from the E. coli Ril or pLysS strains. Furthermore, it had the highest oviposition attraction (49.1%, P < 0.05). These data suggest that biologically active recombinant BinA and BinB toxins might be useful in mosquito control programs, delivered by inactivated bacterial cells or in traps. [ABSTRACT FROM AUTHOR]

Bacillus sphaericus is a bio-control agent effective against Culex quinquefasciatus, the vector of bancroftian filariasis. Apart from its larvicidal effect, there are reports of reduced infection of filarial parasites in mosquitoes exposed to it. In the present study, adults of Cx. quinquefasciatus emerged from B. sphaericus treated larvae were fed on blood samples positive for microfilariae of Wuchereria bancrofti and examined at various time intervals to assess the infection level. The rate of infection was reduced from 95% on day 1 post-feeding to 75% on day 13, when fed with blood sample containing 41 mf/20 μl. The mean parasite burden was also reduced from 4.9 per mosquito on day 1 to 2.15 on day 13. When fed with another sample (30 mf/20 μl), the infection was reduced from 100% on day 1 to 80% on day 13. Reduction in parasite burden was 4.0 to 1.75. Abnormally developed second-stage larvae of the parasite were seen in treated mosquitoes. Thus, the results indicated adverse effect of B. sphaericus treatment on infection and development of the filarial parasite in mosquitoes. The possible reason for the parasite regulation was studied through the assessment of the carryover of the bacterium as well as its toxins to the surviving mosquitoes. The presence of B. sphaericus was determined through plating of homogenate of survived mosquitoes on NYSM agar. Toxic protein was detected through immunoblotting. The bacterium as well as its 41.9-kDa toxic protein was found to be transmitted from larvae to adults and affected the parasite development, directly by the toxin or indirectly by eliciting humoral immune response of the mosquito. [ABSTRACT FROM AUTHOR]

2,4-Dinitroanisole (DNAN) is a low sensitive melt-cast chemical being tested by the Military Industry as a replacement for 2,4,6-trinitrotoluene (TNT) in explosive formulations. Little is known about the fate of DNAN and its transformation products in the natural environment. Here we report aerobic biotransformation of DNAN in artificially contaminated soil microcosms. DNAN was completely transformed in 8 days in soil slurries supplemented with carbon and nitrogen sources. DNAN was completely transformed in 34 days in slurries supplemented with carbons alone and persisted in unamended microcosms. A strain of Bacillus (named 13G) that transformed DNAN by co-metabolism was isolated from the soil. HPLC and LC-MS analyses of cell-free and resting cell assays of Bacillus 13G with DNAN showed the formation of 2-amino-4-nitroanisole as the major end-product via the intermediary formation of the arylnitroso (ArNO) and arylhydroxylamino (ArNHOH) derivatives, indicating regioselective reduction of the ortho-nitro group. A series of secondary reactions involving ArNO and ArNHOH gave the corresponding azoxy- and azo-dimers. Acetylated and demethylated products were identified. Overall, this paper provides the evidence of fast DNAN transformation by the indigenous microbial populations of an amended soil with no history of contamination with explosives and a first insight into the aerobic metabolism of DNAN by the soil isolate Bacillus 13G. [ABSTRACT FROM AUTHOR]

Decolourization, detoxification and biotransformation of 4-chloro-2-nitrophenol (4C2NP) by Bacillus sp. strain MW-1 were studied. This strain decolorized 4C2NP only in the presence of an additional carbon source. On the basis of thin layer chromatography (TLC), high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS), 4-chloro-2-aminophenol, 4-chloro-2-acetaminophenol and 5-chloro-2-methylbenzoxazole were identified as metabolites. Resting cells depleted 4C2NP with stoichiometric formation of 5-chloro-2-methyl benzoxazole. This is the first report of the formation of 5-chloro-2-methylbenzoxazole from 4C2NP by any bacterial strain. [ABSTRACT FROM AUTHOR]

In this study, EPS produced by five Bacillus spp. strains was determined. The only one strain ( B. sphaericus 7055) was selected due to its high EPS production and it was investigated by growing this strain in LB broth medium containing various carbon sources. The highest EPS production of this strain was found in medium containing fructose. However, the effect of different concentrations of fructose and molasses on EPS production by the strain was studied. The maximum EPS yield of the strain 7055 was recorded with 2.5% (w/v) fructose, also the highest EPS production was found in 2.5% (w/v) molasses. The strain 7055 was found to contain (98.6%) galactose and (1.4%) glucuronic acid in control medium whereas the composition of the strain 7055 (2.5% (w/v) fructose) was found to be (99.9%) neutral sugar and (0.1%) Glucuronic acid while the strain 7055 (2.5% (w/v) molasses) was found to contain (65.9%), neutral sugars and (34.1%) glucuronic acid. [ABSTRACT FROM AUTHOR]

The efficacy of whole plant extracts of Leucas aspera and Bacillus sphaericus has been proven against larvicidal and pupicidal activities of the malarial vector, Anopheles stephensi. The present study investigated the larvicidal and pupicidal activity against the first to fourth instar lavae and pupae of the laboratory-reared mosquitoes, A. stephensi. The medicinal plants were collected from the area around Maruthamalai hills, Coimbatore, Tamil Nadu, India. L. aspera whole plant was washed with tap water and shade dried at room temperature. The dried plant materials were powdered by an electric blender. From the powder, 100 g of the plant materials was extracted with 300 ml of organic solvents of ethanol for 8 h using a Soxhlet apparatus. The extracts were filtered through a Buchner funnel with Whatman number 1 filter paper. The crude plant extracts were evaporated to dryness in a rotary vacuum evaporator. The plant extract showed larvicidal and pupicidal effects after 24 h of exposure. All larval instars and pupae have considerably moderate mortality; however, the highest larval mortality was the ethanolic extract of whole plant L. aspera against the first to fourth instar larvae and pupae values of LC = I instar was 9.695%, II instar was 10.272%, III instar was 10.823%, and IV instar was 11.303%, and pupae was 12.732%. B. spaericus against the first to fouth instar larvae and pupae had the following values: I instar was 0.051%, II instar was 0.057%, III instar was 0.062%, IV instar was 0.066%, and for the pupae was 0.073%. No mortality was observed in the control. The present results suggest that the ethanolic extracts of L. aspera and B. sphaericus provided an excellent potential for controlling of malarial vector, A. stephensi. [ABSTRACT FROM AUTHOR]

Integrated vector control is an effective and essential part of any successful vector control program. Increasing insecticide resistance requires strategies to prolong the use of highly effective vector control compounds. Synergistic activity between current effective pesticides is a powerful tool and is one such strategy. In the current study, Cyt1A from Bacillus thurigiensis subsp. israelensis and Bacillus sphaericus B101H5aH5b Bs were evaluated for the synergistic activity against B. sphaericus-resistant strains of Anopheles stephensi. The combinations of Cyt1A and B. sphaericus were found to act synergistically and were greatly enhanced at different ratios. A ratio of 1:4 of Cyt1A and Bs was 13,200-fold more toxic at LC against the Bs-resistant strains of A. stephensi than was Bs alone and this high level of activity resulted from synergism between Cyt1A toxin and Bs. Our results therefore suggest that Cyt1A may enhance toxicity by facilitating the binding or insertion of the binary toxins to the microvillar membrane. [ABSTRACT FROM AUTHOR]

To isolate a salt tolerant hydrogen-producing bacterium, we used the sludge from the intertidal zone of a bathing beach in Tianjin as inoculum to enrich hydrogen-producing bacteria. The sludge was treated by heat-shock pretreatment with three different temperature (80, 100 and 121°C) respectively. A hydrogen-producing bacterium was isolated from the sludge pretreated at 80°C by sandwich plate technique and identified using microscopic examination and 16S rDNA gene sequence analysis. The isolated bacterium was named as Bacillus sp. B2. The present study examined the hydrogen-producing ability of Bacillus sp. B2. The strain was able to produce hydrogen over a wide range of initial pH from 5.0 to 10.0, with an optimum at pH 7.0. The level of hydrogen production was also affected by the salt concentration. Strain B2 has unique capability to adapt high salt concentration. It could produce hydrogen at the salt concentration from 4 to 60‰. The maximum of hydrogen-producing yield of strain B2 was 1.65 ± 0.04 mol H/mol glucose (mean ± SE) at an initial pH value of 7.0 in marine culture conditions. Hydrogen production under fresh culture conditions reached a higher level than that in marine ones. As a result, it is likely that Bacillus sp. B2 could be applied to biohydrogen production using both marine and fresh organic waste. [ABSTRACT FROM AUTHOR]

Production of antimicrobial activity by Bacillus sp. P11 was tested on different byproducts of food industry, like fish meal, grape waste, an industrial fibrous soybean residue, soybean meal, and cheese whey. Bacillus sp. P11 produced the higher amount of antimicrobial activity on soybean meal, followed by fish meal and fibrous soybean residue. Soybean meal was the selected medium to determine the effect of three variables (temperature, initial pH, and substrate concentration) on bacteriocin activity by response surface methodology, using a 2 factorial design. Statistical analysis showed good adequacy to the model ( R of 0.8268). In the range studied, temperature and initial pH of the medium have a significant effect on bacteriocin production, and substrate concentration has no significant effect. Response surface data showed maximum bacteriocin production at initial pH between 7.0 and 8.5 and temperature between 39 and 42 °C. In the optimum conditions (initial pH 7.0 and 42 °C), production of bacteriocin activity by Bacillus sp. P11 was compared using a commercial medium (BHI broth) and soybean meal. Maximum activity achieved with the soybean meal-based medium was similar to that obtained with BHI, indicating that soybean meal may be a cost-effective substrate for production of antimicrobial activity by Bacillus sp. P11. [ABSTRACT FROM AUTHOR]

The main toxicity mechanism of Lysinibacillus sphaericus, which is used in the control of mosquitoes, is its binary toxin produced during sporulation; additionally the Mtx1, Mtx2 and Mtx 3 toxins are expressed in vegetative cells. Mosquito larvicidal potency of the S-layer protein that is expressed in vegetative cells has been determined. The protein is similar to other S-layer proteins of mosquitocidal L. sphaericus strains. The LC50 values of the S-layer protein of the L. sphaericus OT4b25, OT4b26, and III(3)7 strains against third-instar larvae of Culex quinquefasciatus were 8.7, 24 and 0.68 μg/ml, respectively. To our knowledge this is the first study showing the mosquito larvicidal potency of the S-layer protein from Lysinibacillus sphaericus. [ABSTRACT FROM AUTHOR]

Microbial pest control agents or entomopathogens have been considered an interesting alternative to use instead of chemical insecticides. Knowledge of ecotoxicity data is very important to predict the hazard of any product released in the environment and subsidize the regulation of these products by governmental agencies. In the present study four new Brazilian strains of Bacillus and one fungus were tested to evaluate their acute toxicity to the microcrustacean Daphnia similis, the snail Biomphalaria glabrata and the dung beetle Digitonthophagus gazella. The microcrustaceans and the snails were exposed to entomopathogens in synthetic softwater and the beetles were exposed directly in cattle dung. Obtained data reveal low susceptibility of the non-target species to tested microorganisms, with lethal concentrations being observed only at much higher concentrations than that effective against target insects. These results show that the tested strains are selective in their action mode and seem to be non-hazardous to non-target species. [ABSTRACT FROM AUTHOR]

Sustained-release biopolymer based formulations for an indigenous Bacillus sphaericus Neide ISPC-8 strain were developed and evaluated under laboratory conditions. These formulations were assessed for residual activity, response to UV-B radiation and shelf-life. The mosquitocidal activity of tested formulations showed a varied residual activity pattern against Culex quinquefasciatus Say larvae. The residual activity of unformulated spore-crystal suspension drastically decreased after 4 days whereas the formulated preparations persisted until 28 days. All the formulations except sodium alginate-based formulation showed more than 95% mortality within 48 h. Although the sodium alginate-based formulation showed delayed initial mortality, it persisted for a longer period of time (42 days). Based on the persistence studies, promising formulations based on sodium alginate, gelatin and acacia gum were selected for further evaluation. The respective LC values of these three formulations were 0.085, 0.117, and 0.196 mg/l against C. quinquefasciatus after 48 h. The biopolymers combined with Congo red protected the spore-crystal moiety from UV-B radiation. These formulations were found to be stable up to 18 months when stored at room temperature. Thus, these biopolymers could serve as suitable ingredients for development of biopesticide based on B. sphaericus ISPC-8. [ABSTRACT FROM AUTHOR]

ntipathogenic therapy is an outcome of the quorum-sensing inhibition (QSI) mechanism, which targets autoinducer-dependent virulent gene expression in bacterial pathogens. N-acyl homoserine lactone (AHL) acts as a key regulator in the production of virulence factors and biofilm formation in Pseudomonas aeruginosa PAO1 and violacein pigment production in Chromobacterium violaceum. In the present study, the marine bacterial strain SS4 showed potential QSI activity in a concentration-dependent manner (0.5-2 mg/ml) against the AHL-mediated violacein production in C. violaceum (33-86%) and biofilm formation (33-88%), total protease (20-65%), LasA protease (59-68%), LasB elastase (36-68%), pyocyanin (17-86%) and pyoverdin productions in PAO1. The light and confocal laser scanning microscopic analyses confirmed the reduction of the biofilm-forming ability of PAO1 when treated with SS4 extract. Furthermore, the antibiofilm potential was confirmed through static biofilm ring assay, in which ethyl acetate extract of SS4 showed concentration-dependent reduction in the biofilm-forming ability of PAO1. Thus, the result of this study clearly reveals the antipathogenic and antibiofilm properties of the bacterial isolate SS4. Through 16S rDNA analysis, the strain SS4 was identified as Bacillus sp. (GenBank Accession Number: GU471751). [ABSTRACT FROM AUTHOR]

Bacillus anthracis, the etiological agent of anthrax, is responsible for a serious and often fatal disease of mammalian livestock and humans and is an important biological warfare agent. Bacillus sp. AKG was isolated from a hot spring in western Himalayas and species-specific primers targeting gyrB gene identified the strain as B. anthracis within cereus-group. Cloning, sequencing, and phylogenetic analysis of the partial gyrB sequence from strain AKG indicated a close affiliation with B. anthracis and a few recently isolated strains of B. thuringiensis (e.g., strain Al Hakam and serovar konkukian). Phylogenetic analysis of two other housekeeping genes, clpC and gdpD yielded similar results. This observation is further substantiated by phylogenetic reconstruction using concatenated sequences (1680 bases) of the three genes ( gyrB, clpC, and gdpD). Phenotypic features indicated a non-anthracis affiliation for the strain AKG. A novel strategy to distinguish among strains of B. anthracis, B. cereus, and B. thuringiensis based on whole proteome comparison was developed and tested for the identification of this environmental strain. Proteome comparison was used to establish the identity of this unknown environmental strain. Group of replicate 2DE gels for whole cell proteome were generated for each of the three species and strain AKG. Protein spots unique to each group and those showing match between the groups, in a pair-wise comparison, indicated strain AKG as a member of B. thuringiensis. This strategy can be used to assign strains of B. cereus group to their respective species. [ABSTRACT FROM AUTHOR]

Following observations by a service engineer (mid-2007), 12 of the hundreds of installed packaged plant (PP) sewage systems were producing relatively large numbers of Culex quinquefasciatus (Say) and a variety of filth flies in George Town, Grand Cayman. Due to the vector potential of C. quinquefasciatus in the Caribbean, we explored the efficacy of industry-standard insecticides against C. quinquefasciatus. Since PP systems contain an organic biofilm on the water surface, we devised a modified technique for collecting immatures and for the application of insecticides. We treated functioning and non-functioning PP systems with Bacillus sphaericus (VectoLex® WDG), Bacillus thuringiensis var. israelensis (Bti; VectoBac® CG), temephos (Abate® 4-E), monomolecular film or ethyoxylated alcohol surfactant (Agnique® MMF) and (S)-methoprene (Altosid® Pro-G). B. thuringiensis var. israelensis gave control for 35 and 48 days for functioning and non-functioning systems, respectively, with less control from B. sphaericus (high label rate) (c. 25 and 35 days) and temephos (suggested label rate) (c. 27 and 33 days). B. sphaericus (low label rate), monomolecular film and (S)-methoprene gave limited control of immatures (i.e. no mortality detected after 24 h). Treatment options for PP systems are discussed with regard to economics, operational feasibility, and within the context of integrated mosquito management. Functioning systems were recolonized at a faster rate than those that were not functioning, but the level of infestation in the non-functioning systems was considerably greater. [ABSTRACT FROM AUTHOR]

method has been developed for the preconcentration of Th prior to its determination by UV-vis spectrophotometry after complexation with Arsenazo III. A column was packed with Amberlite XAD-4 resin containing immobilized Bacillus sp. as a solid phase extractant. Parameters such as pH of the solution and sample, the concentration of Th, the volume and type of eluent, flow rate and effect of matrix interference were studied. The optimum pH for the sorption of Th is 5.0, and quantitative eluation occurs with 1 mol L HCl. The tolerance limits of some electrolytes were also studied. The loading capacity was determined to be 17.2 mg g. The method was applied to the determination of Th in real ore sample and validated by analyzing a certified reference samples (NCS DC-73350; poplar leaves). [ABSTRACT FROM AUTHOR]

Mosquitocidal toxin 2 (Mtx2) is a mosquito-larvicidal protein produced during vegetative stage of Bacillus sphaericus. The toxin consists of 292 amino acids and has a molecular weight of 31.8 kDa. To determine the active core region of the toxin, amino acids at N- and C-termini were sequentially removed. Deletion up to 23 amino acids from the N-terminus (Met1-Tyr23) did not significantly affect protein production and the toxin activity, whereas removal of 26 amino acids from the N-terminus (Met1-Lys26) completely abolished toxicity even though the protein production remained unchanged. Deletion of only 5 amino acids from the C-terminal end yielded the protein that could not be solubilized and rendered the toxin inactive. The results demonstrated that the C-terminal end of Mtx2 is required for proper folding and toxicity. Amino acids at the N-terminus up to Tyr23 did not play a significant role in protein production and toxicity whereas amino acids between Thr24 and Lys26 are required for full toxicity. [ABSTRACT FROM AUTHOR]

The aim of this work was to statistically optimize the cultural and nutritional parameters for the production of polyhydroxybutyrate (PHB) under submerged fermentation using jackfruit seed hydrolysate as the sole carbon source. On the basis of results obtained from “one variable at a time” experiment, inoculum age, jackfruit seed hydrolysate concentration, and pH were selected for response surface methodology studies. A central composite design (CCD) was employed to get the optimum level of these three factors to maximize the PHB production. The CCD results predicted that jackfruit seed hydrolysates containing 2.5% reducing sugar, inoculum age of 18 h, and initial medium pH 6 could enhance the production of PHB to reach 49% of the biomass (biomass 4.5 g/l and PHB concentration 2.2 g/l). Analysis of variance exhibited a high coefficient of determination ( R) value of 0.910 and 0.928 for biomass and PHB concentration, respectively, and ensured that the quadratic model with the experimental data was a satisfactory one. This is the first report on PHB production by Bacillus sphaericus using statistical experimental design and RSM in submerged fermentation with jackfruit seed hydrolysate as the sole source of carbon. [ABSTRACT FROM AUTHOR]

Four different bacterial strains were isolated from pulp and paper mill sludge in which one alkalotolerant isolate (LP1) having higher capability to remove color and lignin, was identified as Bacillus sp. by 16S RNA sequencing. Optimization of process parameters for decolorization was initially performed to select growth factors which were further substantiated by Taguchi approach in which seven factors, % carbon, % black liquor, duration, pH, temperature, stirring and inoculum size, at two levels, applying L-8 orthogonal array were taken. Maximum color was removed at pH 8, temperature 35°C, stirring 200 rpm, sucrose (2.5%), 48 h, 5% (w/v) inoculum size and 10% black liquor. After optimization 2-fold increase in color and lignin removal from 25-69% and 28-53%, respectively, indicated significance of Taguchi approach in decolorization and delignification of lignin in pulp and paper mill effluent. Enzymes involved in the process of decolorization of effluent were found to be xylanase (54 U/ml) and manganese peroxidase (28 U/ml). Treated effluent was also evaluated for toxicity by Comet assay using Saccharomyces cerevisiae MTCC 36 as model organism, which indicated 58% reduction after treatment by bacterium. [ABSTRACT FROM AUTHOR]

Plant growth-promoting bacteria (PGPB) promote plant growth through a combination of mechanisms which affect mineral nutrition and root development. In this study, we profiled the transcripts in oil palm roots inoculated with a PGPB, Bacillus sphaericus UPMB10, upon the establishment of a stable inoculum on the root surface by cDNA microarray analysis. A total of 70 from more than 3,700 transcripts examined were found to be significantly upregulated in the roots of oil palm inoculated with B. sphaericus UPMB10 compared to those from un-inoculated oil palm roots. Majority of these genes were related to protein synthesis and processing, stress, primary metabolism, and membrane transport. Many ribosomal proteins were upregulated in the roots of B. sphaericus-inoculated oil palm plantlets. In addition, B. sphaericus UPMB10 were able to upregulate the expression of a gene encoding low affinity nitrate transporter, independent of root morphogenesis. These rhizosphere bacteria were also demonstrated to induce stress-related proteins that may lead to tolerance to biotic and abiotic stresses. [ABSTRACT FROM AUTHOR]

Bacillus sp. GHA1 was isolated from water samples and screened for the production of α-amylase. Maximum production of amylase by this strain occurs at 42 °C, pH 6.5 and 72 h after cultivation in production medium. The enzyme was purified through successive applications of ammonium sulfate precipitation, ion exchange and hydrophobic interaction chromatography, resulting in a single band with an apparent molecular weight of 66 kDa, as judged by SDS-PAGE. Calcium analysis of the purified enzymerevealed that it contained three metal ions per molecule. The new extracellular α-amylase is active in a wide range of pH with its maximum activity at pH values 5.5-8.0. The optimum temperature for enzyme activity is 57 ° C and the presence of calcium has relatively low influence on its activity and thermostability. The Bacillus sp. GHA1 a-amylase with these properties may be suitable for use in detergent and food industries. [ABSTRACT FROM AUTHOR]

The effects of rhizobacteria inoculation in modified MS medium containing boron (1 and 10 μM) on the biochemical components, physiological characteristics and mineral content of the in vitro banana plantlets were carried out. The presence of rhizobacteria in the medium supplemented with boron at two concentrations: 1 and 10 μM resulted in an improvement in growth and root biomass compared to the control (uninoculated). Rhizobacteria inoculation also produced an increase in protein, nitrate, soluble nitrogen and chlorophyll contents of the plantlets cultured in MS modified medium containing boron. An increase in percentage of growth (>295%) was shown when boron was applied into medium inoculated with Bacillus sphaericus UPMB10. The effectiveness of inoculation is increased when associated with boron, nitrogen or carbon into the medium. Thus, these bacterial strains could be used as a bioenhancer for growth of in vitro banana plantlets. [ABSTRACT FROM AUTHOR]

Abstract  In this study, Bacillus sphaericus NRC 69 was grown in culture media, in which 12 agricultural wastes were tested as the main carbon, nitrogen and energy sources under solid state fermentation. Of the 12 tested agricultural by-products, wheat bran was the most efficient substrate for the production of B. sphaericus mosquitocidal toxins against larvae of Culex pipiens (LC50 1.2 ppm). Mixtures of tested agricultural wastes separately with wheat bran enhanced the produced toxicity several folds and decreased LC50 between 3.7- and 50-fold in comparison with that of agricultural wastes without mixing. The toxicity of B. sphaericus grown in wheat bran/rice hull at 8/2 (g/g) and wheat bran/barley straw at 1/4 (g/g) showed the same toxicity as that in wheat bran medium (LC50 decreased 17- and 16-fold, in comparison with that in rice hull or barely straw media, respectively). In wheat bran medium, the maximum toxicity of the tested organism obtained at 50% moisture content, inoculum size 84 × 106 CFU/g wheat bran and incubation for 6 days at 30°C. Addition of cheese whey permeate at 10% to wheat bran medium enhanced the toxicity of B. sphaericus NRC 69 about 46%. [ABSTRACT FROM AUTHOR]

Batch and fed-batch fermentation processes were employed to culture an alkalophilic Bacillus sp. for the production of cyclodextrin glucanotransferase (CGTase). CGTase production was repressed by glucose and induced by soluble starch. By fed-batch fermentation, a CGTase activity up to 56 unit ml−1 with 65 g dry cells l−1 were achieved. The CGTase activity and cell density were increased 360 and 510%, respectively, from those values achieved with batch fermentation. [ABSTRACT FROM AUTHOR]

The environmental toxicology of chemical pesticides have increased interest in the development and use of microbial pest control agents. In the present study four new Brazilian strains of Bacillus and one fungus were tested to evaluate the acute oral toxicity and clearance of these microbials in C57BL6 mice. No mortality was observed after exposure for any of the microorganisms tested. Clearance was significant after 30 days but for one strain of B. thuringiensis and one of B. sphaericus this time was not enough to completely eliminate the spores. [ABSTRACT FROM AUTHOR]

Mtx1 and Mtx2 are mosquitocidal toxins produced by some strains of Bacillus sphaericus during vegetative phase of growth. Mtx1 from B. sphaericus 2297 shows higher toxicity against Culex quinquefasciatus larvae than to Aedes aegypti larvae whereas Mtx2 from B. sphaericus 2297 shows lower toxicity against C. quinquefasciatus than to A. aegypti larvae. To test synergism of these toxins against A. aegypti larvae, mtx1 and mtx2 genes were cloned into a single plasmid and expressed in Escherichia coli. Cells producing both Mtx1 and Mtx2 toxins exhibited high synergistic activity against A. aegypti larvae approximately 10 times compared to cells expressing only a single toxin. Co-expression of both toxins offers an alternative to improve efficacy of recombinant bacterial insecticides. There is a high possibility to develop these toxins to be used as an environmentally friendly mosquito control agent. [ABSTRACT FROM AUTHOR]