Decolourization of azo dye using a batch bioreactor by an indigenous bacterium Enterobacter aerogenes ES014 from the waste water dye effluent and toxicity analysis.

Effluents of textile industries caused serious environmental problem throughout the world. In this study, a total of 23 bacterial strains from five bacterial species were isolated from the dye effluent. Of these strains, a unique and novel Enterobacter aerogenes ES014 was utilized for dye decolourization and toxicity analysis. The selected strain could effectively decolourize three selected azo dyes. It showed the capability for decolourizing acid orange (82.3 ± 3.6%), methyl orange (78.2 ± 3.3%), and congo red (81.5 ± 3.2%). The selected bacterial strain significantly decolourized 100 mg/L acid orange at 35 °C, pH 7.5 with 6% sodium chloride concentration. Most of the tested nitrogen and carbon sources effectively enhanced decolourization process. It showed the ability to decolourize acid orange in the culture medium containing 1.5% glucose (100 ± 2.8%) and 0.8% beef extract (100 ± 3.1%). A laboratory-scale batch bioreactor was used to decolourize azo dye at optimized culture conditions. The decolourizing ability improved with 100 mL/h hydraulic retention time. The treated wastewater quality was improved due to sharp depletion of Total Dissolved Solids (TDS), pH, Chemical Oxygen Demand (COD), alkalinity and sulphate concentration. The selected bacteria has the potential to produce dye degrading laccase. Laccase was detected during fermentation process in batch bioreactor as a key enzyme for decolourization produced by E. aerogenes ES014. Phytotoxicity and acute toxicity analysis were performed using Arachis hypogaea (pea nut) seed and first instar larvae of Artemia parthenogenetica (brine shrimp). The seed germination rate of treated wastewater was improved (94.3 ± 1.8%) and enhanced survival rate (91.7 ± 2.9%) in the first instar Artemia larvae treated with wastewater after 24 h. Overall, E. aerogenes ES014, might be a promising bacterial strain for the treatment of textile effluents with high azo dye concentrations.

Biofilm producing indigenous bacteria isolated from municipal sludge and their nutrient removal ability in moving bed biofilm reactor from the wastewater.

In the present study, improved moving bed biofilm reactor (MBBR) was applied to enhance the nutrient removal ability of the municipal wastewater. A total of 18 indigenous bacterial isolates were screened from the sewage sludge sample and nitrate reductase, nitrite reductase and hydroxylamine oxidase was analyzed. The strains Pseudomonas aeruginosa NU1 and Acinetobacter calcoaceticus K12 produced 0.87 ± 0.05 U/mg and 0.52 ± 0.12 U/mg hydroxylamine oxidase, 1.023 ± 0.062 U/mg and 1.29 ± 0.07 U/mg nitrite reductase, and 0.789 ± 0.031 U/mg and 1.07 ± 0.13 U/mg nitrate reductase. Nitrogen and phosphate removal improved by the addition of nutrient sources and achieved > 80% removal rate. pH and temperature of the medium also affected nutrient removal and improved removal was achieved at optimum level (p < 0.05). MBBR was designed with R1 (aerobic), R2 and R3 (anoxic) reactors. MBBR reactors removed acceptable level phosphorus removal properties up to 7.2 ± 3.8%, 42.4 ± 4.6%, and 84.2 ± 13.1% in the R1, R2, R3 and R4 reactors, respectively. Denitrification rate showed linear relationship at increasing concentrations nitrogen content in the reactor and denitrification rate was 1.43 g NO2-N /m2/day at 1.5 g NO2-N /m2/day. Dehydrogenase activity was assayed in all reactors and maximum amount was detected in the aerobic biofilm reactor. Based on the present findings, MBBRs and the selected bacterial strains are useful for the degradation domestic wastewater with minimum working area.

In vitro fibrinolytic activity of an enzyme purified from Bacillus amyloliquefaciens strain KJ10 isolated from soybean paste.

A fibrinolytic protease secreting producing Bacillus amyloliquefaciens strain KJ10 was initially screened from the fermented soybean. Maximum productivity was obtained in the culture medium after 40 h incubation, 34 °C incubation temperature at pH 8.0. Fibrinolytic protease production was enhanced in the culture medium with 1% sucrose (3712 ± 52 U/mL), 1% (w/v) yeast extract (3940 ± 28 U/mL) and 0.1% MgSO4 (3687 ± 38 U/mL). Enzyme was purified up to 22.9-fold with 26%recovery after Q-Sepharose HP column chromatography. After three steps purification, enzyme activity was 1606U/mg and SDS-PAGE analysis revealed 29 kDa protein and enzyme band was detected by zymograpy. Enzyme was highly active at pH 8.0, at wide temperature ranges (40 °C - 55 °C) and was activated by Mn2+ (102 ± 3.1%) and Mg2+ (101.4 ± 2.9%) ions. The purified fibrinolytic enzyme was highly specific against N-Suc-Ala-Ala-Pro-Phe-pNA (189 mmol/min/mL) and clot lytic activity reached 28 ± 1.8% within 60 minin vitro. The purified fibrinolytic enzyme showed least erythrocytic lysis activity confirmed safety to prevent various health risks, including hemolytic anemia. Based on this study, administration of fibrinolytic enzyme from B. amyloliquefaciens strain KJ10 is safe for clinical applications.

Characterization of biofilm formed by multidrug resistant Pseudomonas aeruginosa DC-17 isolated from dental caries.

The present work reports with the screening of biofilm-producing bacteria from the dental caries. The dental pathogens showed resistance against various antibiotics and biofilm forming ability at various levels. Among the bacterial strain, Pseudomonas aeruginosa DC-17 showed enhanced biofilm production. Extracellular polymeric substance (EPS) was synthesized by the selected bacterial isolate considerably and contributed as the major component of biofilm. EPS composed of eDNA, proteins and lipids. The total protein content of the EPS was found to be 1.928 mg/mL and was the major component than carbohydrate and DNA. Carbohydrate content was 162.3 mg/L and DNA content of EPS was 4.95 µg/mL. These macromolecules interacted in the matrix to develop dynamic and specific interactions to signalling biofilm to differentiating various environments. Also, the isolated bacteria showed resistant against various commercially available antibiotics. The isolates showed more resistance against penicillin (98%) and were sensitive against amoxicillin. Among the factors, temperature, pH and sugar concentration influenced biofilm formation. Biofilm forming ability of the selected bacterial stain was tested at various pH values and alkaline pH was favoured for biofilm production. Biofilm production was found to be maximum at 40 °C and 8% sucrose enhanced biofilm formation. Biofilm formed by P. aeruginosa DC-17 was resistant against various tested antimicrobials and chemicals.

Paecilomyces sp. ZB is a cell factory for the production of gibberellic acid using a cheap substrate in solid state fermentation.

Gibberellic acid from the fungi has been widely used in agriculture. In this study, more than 20 fungal isolates were screened and Paecilomyces sp. ZB shown to produce more gibberellic acid than other fungal isolates. Cow dung was used as low cost substrate for gibberellic acid production in solid state fermentation (SSF). Carbon, nitrogen and ionic sources stimulated gibberellic acid production in SSF. Lactose emerged as the significant carbon source supporting more gibberellic acid production (731 µg/g). Among the nitrogen sources, glycine appeared to influence the production of more gibberellic acid (803 µg/g). The process parameters were optimized to enhance gibberellic acid production using a two-level full factorial design and response surface methodology. The amount of gibberellic acid production was influenced mainly by moisture and pH of the substrate. Gibberellic acid production was 1312 µg/g under the optimized conditions and the predicted response was 1339 µg/g. The gibberellic acid yield increased twofolds after medium optimization. The extracted gibberellic acid was sprayed on the growing Mung bean plant and it stimulated the growth of the plant effectively. To conclude, cow dung is a new alternative to produce gibberellic acid in SSF.