Surface modified zeolite, a novel carrier material for Azotobacter chroococcum

A new immobilization matrix based on zeolite has been developed to immobilize Azotobacter chroococcum, for fixing nitrogen, with an intention to hold the cells in the root zone of the plants and to protect them under stressful conditions. The matrix has been developed by modifying the surface of the zeolite with surfactant. This enhances the hydrophobicity of the material and also modifies the surface charge, which in turn enhances the immobilization. Surface modified zeolite-A (SMZ-A) has been compared with commercial zeolite-A (CZA) for immobilization efficiency. CZA is non-toxic for A. chroococcum but is inefficient to adsorb the cells whereas SMZ-A showed 100% adsorption of the microbial cells wherein it was observed that for 1 l of broth culture with total viable count of 10⁸ cfu ml⁻¹ cells of A. chroococcum, a minimum dose of 0.7 g SMZ-A and minimum contact time of 10 h is required to achieve 100% adsorption. Adsorption was confirmed by the cell count and light as well as scanning electron microscopy. Most importantly, the cells adsorbed on SMZ-A could fix the atmospheric nitrogen up to 13 mg g⁻¹ sucrose consumed, which was comparable with the control (unadsorbed cells), which confirms the survival and nitrogen fixation activity of the bacteria. Responsible Editor: Euan K. James.     

Comparative study on removal of enteric pathogens from domestic wastewater using Typha latifolia and Cyperus rotundus along with different substrates

A comparative study was carried out to evaluate the efficiency of different substrate materials along with macrophytes Typha latifolia and Cyperus rotundus in treating domestic wastewater intended for reuse in agriculture. The study was conducted over a period of 6 months with different retention times, and observations were taken twice per month. One-way analysis of variance and Tukey's Honest Significant Difference (HSD) tests were used to determine statistical significant differences between experimental columns. Treatment with T. latifolia planted in sand and mix substrate with 4-day retention time remarkably reduced the concentration of all bacterial pathogens. Log reductions observed were approximately 5.01 and 4.82 for total coliform (TC), 4.46 and 3.93 for Escherichia coli, and 5.52 and 5.48 for Shigella, respectively. Moreover, these treatments were also efficient in completely removing fecal coliform (FC) and Salmonella.Maximum parasites were removed by the treatment having sand alone as a substrate containing C. rotundus, but the difference was not significant from those planted with T. latifolia in the same substrate. The results suggest that T. latifolia aids in bacterial pathogens removal, while C. rotundus aids in parasites removal. Thus, wastewater treatment through constructed wetland having mix plantation of these species along with sand can eliminate some of the major enteric pathogens.     

Kinetics of biosurfactant production by Pseudomonas aeruginosa strain BS2 from industrial wastes

Summary Batch kinetic studies were carried out on rhamnolipid biosurfactant production from synthetic medium, industrial wastes viz. distillery and whey waste as substrates. The results indicated that the specific growth rates ( _max) and specific product formation rates (V _max) from both the wastes are comparatively better than the synthetic medium, revealing that both the industrial wastes (distillery and whey) can be successfully utilized as substrates for biosurfactant production.     

Isolation and characterization of a phorate degrading bacterium

Aims:  To study the degradation of phorate by a bacterium isolated from phorate-contaminated sites.
Methods and Results:  Ralstonia eutropha strain AAJ1 isolated from soil was found to degrade phorate (supplied as sole carbon source) upto 85% in 10 days in liquid medium. Half-life ( t _½) of phorate in the liquid medium in control (uninoculated) and in experimental (inoculated with R. eutropha , strain AAJ1) samples was recorded as 36·49 and 6·29 days, respectively. Kinetics revealed that phorate degradation depends on time and the reaction follows the first order kinetics. Diethyl dithiophosphate was one of the degradation products, which is markedly less toxic than the parent compound; other degradation products included phorate sulfoxide and phorate sulfone. Release of inorganic phosphates and sulfates indicated the potential of the isolate to further degrade the above-mentioned metabolites to simpler forms. The strain was also found to posses phosphomonoesterase and phosphodiesterase enzymatic activity, which are involved in biodegradation of organophosphorus compounds.
Conclusions:  Ralstonia eutropha AAJ1 could degrade and detoxify phorate upto 85% in 10 days in laboratory conditions.
Significance and Impact of the Study:  The isolate has the potential to be utilized for remediation of phorate-contaminated water and soil.     

Adsorption-desorption process using wood-based activated carbon for recovery of biosurfactant from fermented distillery wastewater

Methods used for biosurfactant recovery include solvent extraction, precipitation, crystallization, centrifugation and foam fractionation. These methods cannot be used when distillery wastewater (DW) is used as the nutrient medium for biosurfactant production by Pseudomonas aeruginosa strain BS2, because recovery of biosurfactant by any of these methods imparts color to the biosurfactant. The biosurfactant has a nonaesthetic appearance with lowered surface active properties. These methods cannot be used for continuous recovery of biosurfactant during cultivation. Hence, a new downstream technique for biosurfactant recovery from fermented DW comprised of adsorption-desorption processes using wood-based activated carbon (WAC) was developed. This study involves batch experiments to standardize the factors affecting the rate of biosurfactant adsorption onto WAC. WAC was the most efficient adsorbent among various ones tested (i.e., silica gel, activated alumina and zeolite). The WAC (1% w v(-1)), equilibrium time (90 min), pH range of 5-10 and temperature of 40 degrees C were optimum to achieve 99.5% adsorption efficiency. Adsorption kinetics and intraparticle diffusion studies revealed the involvement of both boundary layer diffusion and intraparticle diffusion. The Langmuir adsorption isotherm of WAC indicated the formation of a monolayer coverage of the biosurfactant over a homogeneous carbon surface, while the Freundlich isotherm showed high adsorption at strong solute concentrations and low adsorption at dilute solute concentrations. WAC concentration of 4% w v(-1) facilitated complete removal of the biosurfactant from collapsed foam (contained 5-fold higher concentration of biosurfactant than was present in fermented DW). Biosurfactant adsorption was of chemisorption type. Acetone (polar solvent) was a specific viable eluant screened among various ones tested because it selectively facilitated maximum recovery, i.e., 89% biosurfactant from WAC. By acetone treatment, complete regeneration of WAC was feasible and WAC can be reused for biosurfactant recovery up to 3 cycles. The recovered biosurfactant showed improved surface-active property (i.e., much lower critical micelle concentration value of 0.013 verses 0.028 mg mL(-1) for biosurfactant recovered by classical methods). The reuse potential of WAC was assessed and results suggest that the carbon can be reused for three consecutive cycles for biosurfactant adsorption from fermented wastewater without any decrease in adsorption efficiency. Thus, this process forms a basis for continuous recovery of biosurfactant from fermented DW and concentrated foam. This process reduces the use of high cost solvent, avoids end product inhibition and minimizes product degradation.     

Microbial population shift caused by sulfamethoxazole in engineered-Soil Aquifer Treatment (e-SAT) system

The engineered-Soil Aquifer Treatment (e-SAT) system was exploited for the biological degradation of Sulfamethoxazole (SMX) which is known to bio-accumulate in the environment. The fate of SMX in soil column was studied through laboratory simulation for a period of 90 days. About 20 ppm SMX concentration could be removed in four consecutive cycles in e-SAT. To understand the microbial community change and biological degradation of SMX in e-SAT system, metagenomic analysis was performed for the soil samples before (A-EBD) and after SMX exposure (B-EBD) in the e-SAT. Four bacterial phyla were found to be present in both the samples, with sample B-EBD showing increased abundance for Actinobacteria, Bacteroidetes, Firmicutes and decreased Proteobacterial abundance compared to A-EBD. The unclassified bacteria were found to be abundant in B-EBD compared to A-EBD. At class level, classes such as Bacilli, Negativicutes, Deltaproteobacteria, and Bacteroidia emerged in sample B-EBD owing to SMX treatment, while Burkholderiales and Nitrosomonadales appeared to be dominant at order level after SMX treatment. Furthermore, in response to SMX treatment, the family Nitrosomonadaceae appeared to be dominant. Pseudomonas was the most dominating bacterial genus in A-EBD whereas Cupriavidus dominated in sample B-EBD. Additionally, the sulfur oxidizing bacteria were enriched in the B-EBD sample, signifying efficient electron transfer and hence organic molecule degradation in the e-SAT system. Results of this study offer new insights into understanding of microbial community shift during the biodegradation of SMX.     

Biodegradation of chrysene by the bacterial strains isolated from oily sludge

The biodegradation studies were conducted to test the ability of the bacterial strains (Chry2 and Chry3) isolated from the oily sludge obtained from Gujarat refinery, India, for utilization of chrysene in the liquid medium. Biodegradation of the compound was confirmed using gas chromatography and the percent degradation was calculated to be 15.0 and 17% by Chry2 and Chry3, respectively. The biodegradation results were supported by increase in viable cell count and dry biomass, in the presence of chrysene as the sole carbon source. Both the cultures produced biosurfactant which was indicated by the reduction in surface tension of the growth medium. Presence of catechol 2, 3-dioxygenase gene in Chry3 indicated its potential for degradation of PAHs through meta cleavage degradation pathway. Both the strains were found to possess catechol 1,2-dioxygenase and catechol 2,3-dioxygenase enzyme activities. Based on morphological and biochemical tests, the cultures were tentatively identified as Bacillus sp. (Chry2) and Pseudomonas sp. (Chry3).     

Degradation of Di- Through Hepta-Chlorobiphenyls in Clophen Oil Using Microorganisms Isolated from Long Term PCBs Contaminated Soil

Present work describes microbial degradation of selected polychlorinated biphenyls (PCBs) congeners in Clophen oil which is used as transformer oil and contains high concentration of PCBs. Indigenous PCBs degrading bacteria were isolated from Clophen oil contaminated soil using enrichment culture technique. A 15 days study was carried out to assess the biodegradation potential of two bacterial cultures and their consortium for Clophen oil with a final PCBs concentration of 100 mg kg⁻¹. The degradation capability of the individual bacterium and the consortium towards the varying range of PCBs congeners (di- through hepta-chlorobiphenyls) was determined using GCMS. Also, dehydrogenase enzyme was estimated to assess the microbial activity. Maximum degradation was observed in treatment containing consortium that resulted in up to 97 % degradation of PCB-44 which is a tetra chlorinated biphenyl whereas, hexa chlorinated biphenyl congener (PCB-153) was degraded up to 90 % by the consortium. This indicates that the degradation capability of microbial consortium was significantly higher than that of individual cultures. Furthermore, the results suggest that for degradation of lower as well as higher chlorinated PCB congeners; a microbial consortium is required rather than individual cultures.     

Immobilization of carbonic anhydrase in alginate and its influence on transformation of CO2 to calcite

Present investigation entails carbonic anhydrase (CA) immobilization and its influence on transformation of CO₂ to calcite. CA enzyme was immobilized in alginate beads, subsequently maintained its catalytic efficiency after sequential operational cycles. The immobilized beads showed better operational stability by retaining nearly 67% of its initial activity even after six cycles. Batch scale studies with free and immobilized enzyme revealed that the entrapped CA hydrates CO₂ to bicarbonate and/or carbonate which was then made to react with Ca²⁺ ions to transform into calcite. Calcite was characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The entrapped CA was employed for the performance evaluation with respect to several operational parameters including the influence of enzyme concentration in free and immobilized condition. It was concluded that immobilized CA in alginate beads would have the potential for CO₂ sequestration by biomimetic route.     

Assessment of hydrocarbon degradation potentials in a plant–microbe interaction system with oil sludge contamination: A sustainable solution

A pot culture experiment was conducted for 90 days for the evaluation of oil and total petroleum hydrocarbon (TPH) degradation in vegetated and non-vegetated treatments of real-field oil-sludge-contaminated soil. Five different treatments include (T1) control, 2% oil-sludge-contaminated soil; (T2), augmentation of microbial consortium; (T3), Vertiveria zizanioides; (T4), bio-augmentation along with V. zizanioides; and (T5), bio-augmentation with V. zizanioides and bulking agent. During the study, oil reduction, TPH, and degradation of its fractions were determined. Physico-chemical and microbiological parameters of soil were also monitored simultaneously. At the end of the experimental period, oil content (85%) was reduced maximally in bio-augmented rhizospheric treatments (T4 and T5) as compared to control (27%). TPH reduction was observed to be 88 and 89% in bio-augmented rhizospheric soil (T4 and T5 treatments), whereas in non-rhizospheric and control (T2 and T1), TPH reduction was 78 and 37%, respectively. Degradation of aromatic fraction after 90 days in bio-augmented rhizosphere of treatments T4 and T5 was found to 91 and 92%, respectively. In microbial (T2) and Vertiveria treatments (T3), degradation of aromatic fraction was 83 and 68%, respectively. A threefold increase in soil dehydrogenase activity and noticeable changes in organic carbon content and water-holding capacity were also observed which indicated maximum degradation of oil and its fractions in combined treatment of plants and microbes. It is concluded that the plant–microbe soil system helps to restore soil quality and can be used as an effective tool for the remediation of oil-sludge-contaminated sites.