The discrepant metabolic pathways of PAHs by facultative anaerobic bacteria under aerobic and nitrate-reducing conditions.

Studies regarding the facultative anaerobic biodegradation of polycyclic aromatic hydrocarbons (PAHs) were still in the initial stage. In this study, a facultative anaerobe which was identified as Bacillus Firmus and named as PheN7 was firstly isolated from the mixed petroleum-polluted soil samples using phenanthrene and nitrate as the solo carbon resource and electron acceptor under anaerobic condition. The degradation rates of PheN7 towards phenanthrene were detected as 33.17 µM/d, 13.81 µM/d and 7.11 µM/d at the initial phenanthrene concentration of 250.17 µM with oxygen, nitrate and sulfate as the electron acceptor, respectively. The metabolic pathways toward phenanthrene by PheN7 were deduced combining the metagenome analysis of PheN7 and intermediate metabolites of phenanthrene under aerobic and nitrate-reducing conditions. Dioxygenation and carboxylation were inferred as the initial activation reactions of phenanthrene degradation in these two pathways. This study highlighted the significance of facultative anaerobic bacteria in natural PAHs biodegradation, revealing the discrepant metabolic fates of PAHs by one solo bacteria under aerobic and anaerobic environments.

Utilization and transformation of Chrysotila dentata-derived dissolved organic matter by phycosphere bacteria Marinobacter hydrocarbonoclasticus and Bacillus firmus.

The dissolved organic matter (DOM) released from the cocoolithophores (Chrysotila dentata) was studied in laboratory experiments after co-culturing C. dentata with bacteria. Marinobacter hydrocarbonoclasticus (CA6)-γ-Proteobacteria and Bacillus firmus (CF2) were used to investigate the utilization and processing of the DOM derived from C. dentata, utilizing fluorescence excitation-emission matrix (EEM) combined with parallel factor analysis (EEM-PARAFAC), while measuring algal abundance and photosynthetic parameters. The experimental groups consisted of axenic C. dentata groups, filter cultured with bacteria (CA6 or CF2) groups, C. dentata co-cultured with bacteria (CA6 or CF2) groups and axenic bacteria (CA6 or CF2) groups. We then evaluated the processing of DOM by determining four fluorescence indices. The number of C. dentata cells and the photosynthetic capacity of microalgae were enhanced by CA6 and CF2. The main known fluorophores, including humic-like components and protein-like components, were present in all sample. The protein-like component of algal-bacterial co-cultures was effectively utilized by CA6 and CF2. The humic-like components increased at the end of the culture time for all cultures. Meanwhile, the average fluorescence intensity of protein-like in CA6 co-culture with algae was lower than that in CF2 co-culture with algae over time. On the other hand, the average fluorescence intensity of humic-like in CA6 was higher than CF2. However, the total change in fluorescence in humic-like and protein-like of axenic CF2 cultures was lower than that of CA6. Hence, the ability of CA6 to transform microalgal-derived DOM was superior to that of CF2, and CF2's ability to consume bacterial-derived DOM was superior to that of CA6.

Bioaugmentation of polycyclic aromatic hydrocarbon (PAH)-contaminated soil with the nitrate-reducing bacterium PheN7 under anaerobic condition.

The remediation of polycyclic aromatic hydrocarbon (PAH)-contaminated soil under anaerobic condition is still a huge challenge. In this study, an anaerobic Bacillus firmus strain named PheN7 was firstly isolated from mixture of contaminated soil and sludge samples with phenanthrene as the sole carbon resource under nitrate reducing environment. The anaerobic strain was then inoculated combining with nitrate into the phenanthrene-spiked PAH-contaminated soil to investigate the remediation efficiency by anaerobic bioaugmentation (BA). Results showed that the synergy between PheN7 and indigenous degrading bacteria promoted the remediation efficiency of soil. The average removal efficiencies of phenanthrene in 56 days were 1.73 mg/kg soil·d in BA group, much higher than biostimulation group (sole nitrate addition) and natural degradation which achieved 1.48 mg/kg soil·d and 1.24 mg/kg soil·d of degradation rate, respectively. The outstanding adaptability of PheN7 made it become the dominant species in soil in the terminal period, but the invasion of PheN7 also resulted in the decline of diversity of the indigenous microbial community. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States 2 (PICRUSt 2) results showed that a series of functional genes encoding anaerobic phenanthrene degradation and nitrate reductase enzymes in soil were remarkably strengthened with the addition of PheN7. This study confirmed the contribution of PheN7 as the anaerobic inoculum in PAH-contaminated soil remediation, further evaluating the practical applicability of anaerobic bioaugmentation technology in on-site remediation of real PAH-contaminated sites.

Bio-control of Stem Rot in Jerusalem Artichoke (Helianthus tuberosus L.) in Field Conditions.

Stem rot is a serious disease in Jerusalem artichoke (JA). To reduce the impact of this disease on yield and quality farmers often use fungicides, but this control method can be expensive and leave chemical residues. The objective of this study was to evaluate the efficacy of two biological control agents, Trichoderma harzianum T9 and Bacillus firmus BSR032 for control of Sclerotium rolfsii under field conditions. Four accessions of JA (HEL246, HEL65, JA47, and JA12) were treated or notreated with T. harzianum T9 and B. firmus BSR032 in a 4 × 2 × 2 factorial experiment in two fields (environments), one unfertilized and one fertilized. Plants were inoculated with S. rolfsii and disease was evaluated at 3-day intervals for 46 days. T. harzianum T9 and B. firmus BSR032 reduced disease incidence by 48% and 49%, respectively, whereas T. harzianum T9 + B. firmus BSR032 reduced disease incidence by 37%. The efficacy of T. harzianum T9 and B. firmus BSR032 for control of S. rolfsii was dependent on environments and genotypes. The expression of host plant resistance also depended on the environment. However, HEL246 showed consistently low disease incidence and severity index in both environments (fertilized and unfertilized). Individually, T. harzianum T9, B. firmus BSR032, or host plant resistance control stem rot caused by S. rolfsii in JA. However, no combination of these treatments provided more effective control than each alone.

Effects of Bacillus firmus e volumine ex muris cellulae 6x on cytolytic activity of natural killer cells in vitro

Natural killer (NK) cells are among the first in defense of the innate immune system by eliminating a variety of abnormal or stressed cells such as cancer cells or virus-infected cells. Individuals who exhibit low cytolytic NK cell activity are believed to be at higher risk of viral infection, tumorigenesis, and various other diseases of the immune system. Therefore, restoration of impaired NK cell function might be an essential step in immunostimulatory therapy of immunocompromised patients. Bacillus firmus is a non-pathogenic gram-positive bacterium of the environment, which possesses various immunomodulatory properties in vitro and in vivo. This retrospective study reports on the effect of B. firmus on the activity of NK cells in vitro. Basal cytolytic NK cell activity against tumor cells among peripheral blood mononuclear cells (PBMCs) of routine patients was determined in a standardized NK cell cytotoxicity assay. The impact of cultivation of PBMCs with B. firmus preparation Bacillus firmus e volumine ex muris cellulae (Bacillus firmus (evc)) 6x on tumor cell killing by NK cells was monitored in relation to basal NK cell activity. This study showed that stimulation of PBMCs with Bacillus firmus (evc) 6x in vitro led to a significant increase in NK cell function. Substantial improvement in cytolytic NK cell activity (more than 1.3-fold of basal activity) was much more pronounced for patients with compromised NK cell function. Due to its immunostimulatory mode of action, Bacillus firmus (evc) may be of particular importance in therapy of patients with NK cell deficiency.

Efficient biological pretreatment and bioconversion of corn cob by the sequential application of a Bacillus firmus K-1 cellulase-free xylanolytic enzyme and commercial cellulases.

We used agricultural residue, corn cob, with biorefinery and bioeconomy concepts. At short-time cultivation in corn cob (12 h), Bacillus firmus K-1 produced cellulase-free xylanolytic enzyme, with xylooligosaccharides (XOSs), X5 and X6, as the main products, which can be used in a variety of applications. The xylanolytic enzyme produced from B. firmus K-1 effectively degraded xylan in corn cob, which was examined by chemical composition, scanning electron microscope (SEM), and Fourier transform infrared spectroscopy (FTIR). After cultivation, the xylan contained in the corn cob residue was decreased (as biological pretreatment), causing morphological and structural changes, including creating porosity and increasing the surface area and the exposure of cellulose of pretreated corn cob. These results lead to an improvement of cellulose access by cellulases. Commercially available cellulases, Accellerase® 1500 and Cellic® CTec2, yielded significantly higher glucose concentrations from pretreated corn cob compared to untreated corn cob. After saccharification, the lignin-rich corn cob residue can be used as a raw material for other purposes. Moreover, the B. firmus cells, with a low risk to human health, can be used in some applications. This study presents an efficient method for producing high-value-added products from agricultural residue (corn cob) through biological processes which are environmentally friendly and economically viable. KEY POINTS: • High-value-added products were efficiently produced from corn cob by B. firmus K-1. • After biological pretreatment by B. firmus K-1, cellulase can better reach cellulose. • XOSs and cellulose-derived glucose were the main products from corn cob.

Identification of Multi-Potent Protein Subtilisin A from halophilic bacterium Bacillus firmus VE2.

Screening of halophiles with antimicrobial activity in saltpan soil samples from Nagapattinam district, Tamil Nadu, revealed isolate VE-2 as the most potent, identified as Bacillus firmus strain VE-2 through 16s rRNA gene sequencing. It had an optimum growth condition (OD 3.1) and antimicrobial protein (AMP) production (450 µg/mL) at 37 °C, pH 8, 25% NaCl, and 36 h incubation. SDS-PAGE analysis of the purified AMP showed the molecular weight of 36 kDa. HPLC analysis of the purified AMP showed different amino acids, such as asparagines, alanine, lysine, proline, threonine, glycine, cysteine, serine, aspartic acid leucine, and valine. Further characterization and identification using FT-IR, 2D-PAGE, MALDI-TOF, and in-silico analysis showed that the isolated AMP had the highest similarity to Subtilisin-A. It showed antibacterial activity against clinical bacterial pathogens like S. aureus, S. pyogenes, C. diphtheria, E. coli, and P. aeruginosa with the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration of 2.5 µg/mL and 20 µg/mL and also against various fungal pathogens such as A. niger, A. flavus, C. albicans, C. tropicalis and C. parapsilosis with the MIC and minimum fungicidal concentrations of 1.25-80 µg/mL. The purified AMP had excellent antioxidant potential, showed a scavenging effect against DPPH and Nitric oxide radicals, and displayed anticancer activity against HeLa cell lines with the IC50 values 53 µg/mL. Hence, the purified bioactive antimicrobial peptides (AMP) could also be used in anticancer therapies.

From Genome to Field-Observation of the Multimodal Nematicidal and Plant Growth-Promoting Effects of Bacillus firmus I-1582 on Tomatoes Using Hyperspectral Remote Sensing.

Root-knot nematodes are considered the most important group of plant-parasitic nematodes due to their wide range of plant hosts and subsequent role in yield losses in agricultural production systems. Chemical nematicides are the primary control method, but ecotoxicity issues with some compounds has led to their phasing-out and consequential development of new control strategies, including biological control. We evaluated the nematicidal activity of Bacillus firmus I-1582 in pot and microplot experiments against Meloidogyne luci. I-1582 reduced nematode counts by 51% and 53% compared to the untreated control in pot and microplot experiments, respectively. I-1582 presence in the rhizosphere had concurrent nematicidal and plant growth-promoting effects, measured using plant morphology, relative chlorophyll content, elemental composition and hyperspectral imaging. Hyperspectral imaging in the 400-2500 nm spectral range and supervised classification using partial least squares support vector machines successfully differentiated B. firmus-treated and untreated plants, with 97.4% and 96.3% accuracy in pot and microplot experiments, respectively. Visible and shortwave infrared spectral regions associated with chlorophyll, N-H and C-N stretches in proteins were most relevant for treatment discrimination. This study shows the ability of hyperspectral imaging to rapidly assess the success of biological measures for pest control.

Whole Genome Sequencing and Comparative Genomics of Two Nematicidal Bacillus Strains Reveals a Wide Range of Possible Virulence Factors.

Bacillus firmus nematicidal bacterial strains are used to control plant parasitic nematode infestation of crops in agricultural production. Proteases are presumed to be the primary nematode virulence factors in nematicidal B. firmus degrading the nematode cuticle and other organs. We determined and compared the whole genome sequences of two nematicidal strains. Comparative genomics with a particular focus on possible virulence determinants revealed a wider range of possible virulence factors in a B. firmus isolate from a commercial bionematicide and a wild type Bacillus sp. isolate with nematicidal activity. The resulting 4.6 Mb B. firmus I-1582 and 5.3 Mb Bacillus sp. ZZV12-4809 genome assemblies contain respectively 18 and 19 homologs to nematode-virulent proteases, two nematode-virulent chitinase homologs in ZZV12-4809 and 28 and 36 secondary metabolite biosynthetic clusters, projected to encode antibiotics, small peptides, toxins and siderophores. The results of this study point to the genetic capability of B. firmus and related species for nematode virulence through a range of direct and indirect mechanisms.

Biodegradation of textile dye Reactive Blue 160 by Bacillus firmus (Bacillaceae: Bacillales) and non-target toxicity screening of their degraded products.

The study was envisioned to evaluate the decolorization of Reactive Blue 160 (RB160) dye by using indigenous microbes. Contaminated soil from textile dye industry was collected from Noyyal river basin, Tamil Nadu, India. Potential dye degrading bacterial strain was recognized as Bacillus firmus by 16SrRNA gene sequencing analysis. RB160 dye (500 µg/ml) was effectively degraded by B. firmus and toxicological analyses were performed with RB160 and their degraded product. Phytotoxicity revealed that degraded product of RB160 into non-toxic nature by B. firms. Toxicity assays were carried out on root cells of Allium cepa and human skin cell line (CRL 1474). Toxicity analysis of A. cepa and cell line signifies that dye exerts toxic cause on the root cells and IC50 values of RB160 showed toxic to human skin cell lines, while degradation products of the dye are moderately less in toxic. Zebrafish embryo toxicity also evaluated by RB160 and degraded product on phenotypic deformation, survival, hatching and heartbeat rate. However, RB160 with concentration of 500 µg/ml decrease in the survival, hatching, heartbeat rate and induced phenotypic alterations. In which, degraded products exhibited significant development in zebrafish embryos as compared to dye. Based on the studies effects of RB160 and capability of B. firmus can effectively degrade RB160, and their degraded products were harmless to the environments and aquatic system.

Movement of seed- and soil-applied fluopyram in soil columns.

The movement of seed- and soil-applied fluopyram was evaluated in soil columns. The nematicide was sampled at three soil depths and used in a nematode motility bioassay. Based on Meloidogyne incognita mortality, the downward movement of soil-applied fluopyram was affected by soil type and application method. No nematode-toxic levels of soil-applied fluopyram were detected past 5 cm depth in sandy loam soil compared to 10 cm depth in sandy soil. A slower rate of water infiltration had little impact on the movement of soil-applied fluopyram in sandy soil, but did affect the movement of soil-applied abamectin. In the seed-applied nematicide experiments, a greater effect on nematode mortality was observed at the 0 to 5 cm depth in sandy soil with fluopyram- than abamectin-treated cotton seed, whereas a similar effect was observed with soybean seed. No effect on nematode motility was observed with other seed-applied nematicides, thiodicarb, and Bacillus firmus. Overall, soil-applied fluopyram had a greater effect on M. incognita mortality at 10 cm depth in sandy soil than seed-applied fluopyram. These data provide a better understanding as to the movement of fluopyram as affected by soil type, water infiltration rates, and application methods.

Bacillus firmus (SW5) augments salt tolerance in soybean (Glycine max L.) by modulating root system architecture, antioxidant defense systems and stress-responsive genes expression.

Soil salinity is an adverse abiotic factor which reduces plant growth, yield and quality. Plant growth-promoting rhizobacteria (PGPR) have a great potential to enhance growth and alleviate saline stress effects without harming the environment via regulating physiological and molecular processes in plants. This study aimed at investigating Bacillus firmus SW5 effects on the performance of soybean (Glycine max L.) subjected to salt stress (0, 40 and 80 mM NaCl). Salinity stress mitigated the growth and biomass yield, root architecture traits, nutrient acquisition, chlorophyll level, transpiration rate (E), photosynthesis rate (Pn), stomatal conductance (gs), soluble proteins content, soluble sugars content and total phenolics and flavonoid contents of soybean plants. High salinity augmented the levels of osmolytes (glycine betaine and proline), hydrogen peroxide (H2O2), malondialdehyde (MDA) and the activities of antioxidant enzymes (APX, CAT, SOD and POD) in soybean plants. High salinity also induced the expression of antioxidant enzyme-encoding genes (APX, CAT, POD, Fe-SOD) and genes conferring tolerance to salinity (GmVSP, GmPHD2, GmbZIP62, GmWRKY54, GmOLPb, CHS) in soybean plants. On the other hand, inoculation of NaCl-stressed soybean plants with Bacillus firmus SW5 promoted the growth and biomass yield, chlorophyll synthesis, nutrient uptake, gas exchange parameters, osmolytes levels, total phenolic and flavonoid contents, and antioxidant enzymes activities, in comparison with the plants treated with NaCl alone. Bacillus firmus SW5 inoculation also significantly reduced the IC50 values for both DPPH and ß-carotene-linoleic acid assays and indicated higher antioxidant activities in salt-stressed plants. Furthermore, contents of H2O2 and MDA were alleviated in salinity-stressed soybean plants inoculated with Bacillus firmus SW5, in comparison with those in plants exposed to NaCl alone. The antioxidant enzyme-encoding genes and stress-related genes exhibited the highest expression levels in soybean plants inoculated with Bacillus firmus SW5 and treated with 80 mM NaCl. Taken together, our results demonstrate the crucial role of Bacillus firmus SW5 in ameliorating the adverse effects of high salinity on soybean growth and performance via altering the root system architecture and inducing the antioxidant defense systems and stress-responsive genes expression.

Different strategies for cyclodextrin production: Ultrafiltration systems, CGTase immobilization and use of a complexing agent.

The study comparatively evaluated diverse strategic models of cyclodextrin (CD) production by the CGTase of Bacillus firmus strain 37: continuous production and repetitive batches in ultrafiltration systems; immobilization of CGTase on curdlan and vegetable sponge natural supports; the use of the glycyrrhizin complexing agent to modulate CGTase selectivity in favor of γ-CD production. All strategies had in common the possibility of separation of CGTase from its inhibitory products and its reuse. In the continuous production model, at 48 h of assay, the highest productivity and selectivity for ß-CD were obtained, 1.47 mmol/L/h and 92.8%, respectively. Glycyrrhizin was able to modulate the production of γ-CD with selectivity of 61.2% for 30-h batches. The comparative evaluation of the different strategic models for obtaining CDs showed particularities that should be considered, and most of the models studied returned satisfactory yields as well as excellent selectivity.

Cloning, purification and characterization of novel Cu-containing nitrite reductase from the Bacillus firmus GY-49.

Nitrite is generated from the nitrogen cycle and its accumulation is harmful to environment and it can be reduced to nitric oxid by nitrite reductase. A novel gene from Bacillus firmus GY-49 is identified as a nirK gene encoding Cu-containing nitrite reductase by genome sequence. The full-length protein included a putative signal peptide of 26 amino acids and shown 72.73% similarity with other Cu-containing nitrite reductase whose function was verified. The 993-bp fragment encoding the mature peptide of NirK was cloned into pET-28a (+) vector and overexpressed as an active protein of 36.41 kDa in the E.coli system. The purified enzyme was green in the oxidized state and displayed double gentle peaks at 456 and 608 nm. The specific activity of purified enzyme was 98.4 U/mg toward sodium nitrite around pH 6.5 and 35 °C. The K m and K cat of NirK on sodium nitrite were 0.27 mM and 0.36 × 103 s-1, respectively. Finally, homology model analysis of NirK indicated that the enzyme was a homotrimer structure and well conserved in Cu-binding sites for enzymatic functions. This is a first report for nitrite reductase from Bacillus firmus, which augment the acquaintance of nitrite reductase.

PROMOCIÓN DEL CRECIMIENTO DE Baccharis macrantha (ASTERACEAE) CON BACTERIAS SOLUBILIZADORAS DE FOSFATOS ASOCIADAS A SU RIZOSFERA / Growth Promotion of Baccharis macrantha (Asteraceae) by Phosphate Solubilizing Rhizosphere Bacteria

El objetivo de esta investigación fue aislar y caracterizar bacterias solubilizadoras de fosfatos (BSF) asociadas a la rizosfera de Baccharis macrantha y Viburnum triphyllum, y evaluar su capacidad para solubilizar fosfatos en condiciones in vitro. Además se determinó el efecto de la inoculación de las cepas de BSF más eficientes sobre el crecimiento de B. macrantha. Las muestras de suelo rizosférico de B. macrantha y V. triphyllum fueron colectadas en los meses de mayo-período de lluvia y septiembre-período seco del 2012. Para la cuantificación de bacterias heterótrofas cultivables y BSF se empleó el método de recuento en placa en los medios Agar Tripticasa de Soya y Pikovskaya (PVK) respectivamente. La capacidad de solubilización de fosfatos de las cepas aisladas se estimó a partir del diámetro de los halos formados alrededor de las colonias en el medio de cultivo PVK después de 7 días de incubación a 28 °C. Los ensayos de inoculación en B. macrantha se realizaron con las BSF más eficientes. La inoculación de las BSF B. firmusy P. fluorescens de forma individual y como inoculante combinado mostro un efecto benéfico, incrementando significativamente el porcentaje de germinación de semillas, la altura de la plántula, la longitud de la raíz y el peso seco de B. macrantha. La inoculación de BSF podría ser considerada una estrategia para mejorar el crecimiento y establecimiento de B. macrantha en pastizales abandonados.

The objectives of this research was to isolate and characterize phosphate solubilizing bacteria (BSF) associated to the rhizosphere of Baccharis macrantha and Viburnum triphyllum, and to assess their ability to solubilize phosphate under conditions in vitro. Furthermore to determine the effect of inoculation of the strains BSF more efficient on the growth of B. macrantha. Rhizosphere soil samples of B. macrantha and V. triphyllum were collected in the months of May-rainy season and September-period dry the 2012. Trypticase Soya Agar and Pikovskaya (PVK) were used for quantification of culturable heterotrophic bacteria and BSF, respectively. The phosphate solubilizing capacity of the isolated strains was estimated from the diameter of the halo around the colonies formed in the culture medium PVK after 7 days incubation at 28 °C. Inoculation assays were performed with more efficient BSF in B. macrantha. Inoculation of BSF Bacillus firmus and Pseudomona fluorescens individually and as inoculant combined showed a beneficial effect, significantly increasing the percentage of seed germination, seedling height, root length and dry weight of B . macrantha. Inoculation the BSF could be considered a strategy to improve the growth and development of B. macrantha in abandoned pastures.

Mixed Cultures Fermentation for the Production of Poly- ß-hydroxybutyrate

Poly-ß-hydroxybutyrate (PHB) is a biodegradable intracellular microbial product produced by many bacteria and it is comparable to some of the petrochemical derived thermoplastics such as polypropylene. One of the main barriers for the commercial exploitation is the high cost of the substrate for the production of biopolymer. The utilization of mixed microbial cultures facilitates the use of complex substrates thereby reducing the cost of PHB production. In the present study, mixed culture systems were evaluated for PHB production. Bacillus firmus NII 0830 was used for the production of PHB since it accumulates a large amount of PHB and a second organism Lactobacillus delbrueckii NII 0925 was used to provide lactic acid. FTIR and 1H NMR analyses revealed that the PHB extracted from pure culture and mixed culture showed exact match to that of standard PHB. Biodegradation studies of the PHB blends showed 87% degradation. It was also found that a consortium of organisms degraded the films faster than a single organism.

Draft genome sequence of Bacillus firmus DS1.

Bacillus firmus DS1, an aerobic, Gram-positive, spore-forming bacterium isolated from marine sediment of the China South Sea coast. Here, the first draft genome sequence of B. firmus DS1 that may help us to clarify the evolutionary status of B. firmus, also will give the opportunity to provide the genetic basis for heavy-metal ion absorption in environmental bioremediation, the enzymes in industrial production and more other active ingredients application.

Quorum quenching activity in cell-free lysate of endophytic bacteria isolated from Pterocarpus santalinus Linn., and its effect on quorum sensing regulated biofilm in Pseudomonas aeruginosa PAO1.

Quorum sensing mechanism allows the microorganisms to resist the antibiotic treatment by forming biofilms. Quorum quenching is one of the mechanisms to control the development of drug resistance in microbes. Endophyte bacteria are beneficial to plant growth as they support the immune system against the pathogen attack. The endophytic bacteria present in Pterocarpus santalinus were screened for the presence of N-acyl homoserine lactones (AHLs) degrading bacteria using biosensor strains and further confirmed by quantifying the violacein production. Cell-free lysate of endophytic bacteria, Bacillus firmus PT18 and Enterobacter asburiae PT39 exhibited potent AHL degrading ability by inhibiting about 80% violacein production in biosensor strain. Furthermore, when the cell-free lysate was applied to Pseudomonas aeruginosa PAO1 and PAO1-JP2 biofilm it resulted in significant (p<0.01) inhibition of biofilm formation. The biofilm inhibition was confirmed by visualization of biofilm slides under fluorescence microscopy, which showed decrease in total biomass formation in treated slides. Isolation and amplification of the gene (aiiA) indicated that the presence of AHL lactonase in cell-free lysate and sequence alignment indicated that AiiA contains a "HXHXDH" zinc-binding motif that is being conserved in several groups of metallohydrolases. Therefore, the study shows the potential of AHLs degradation by AHL lactonase present in cell-free lysate of isolated endophytic bacteria and inhibition of quorum sensing regulated biofilm formation in P. aeruginosa PAO1.

Effects of a Commercial Formulation of Bacillus firmus I-1582 on Golf Course Bermudagrass Infested with Belonolaimus longicaudatus.

One of the primary pests of bermudagrass (Cynodon spp.) on golf courses in the southeastern United States is Belonolaimus longicaudatus (sting nematode). In 2011, a commercial formulation of Bacillus firmus I-1582, Nortica 5WG, was launched in the United States for management of plant-parasitic nematodes on turfgrasses. To test the efficacy of late winter/early spring application of this biopesticide on B. longicaudatus, two field trials in 2009 compared B. firmus with fenamiphos and untreated control treatments. In 2011, two additional field trials compared treatment with B. firmus with untreated control only. These trials measured treatment effects on the population density of B. longicaudatus, turf root length, and turf percent green cover. In all four trials, treatment with B. firmus improved root length and decreased numbers of B. longicaudatus in contrast to the untreated. These results indicate that late winter/early spring application of B. firmus is an effective biopesticide treatment for management of B. longicaudatus on golf course bermudagrass.

ISOLATION AND IDENTIFICATION OF A HIGH-MELANIN-PRODUCING BACTERIUM / 微生物学通报

A bacterium strain BFHM2002 is isolated from Lake Donghu, Wuhan. BFHM2002 has advantages that it can produce melanin with a high rate and high yield in the absence of tyrosine. Induced by tyrosine, melanin yield can be dramatically increased. BFHM2002 may be identified as a new strain in Bacillus firmus, for melanin-production.