Optimization of microbial fuel cell process using a novel consortium for aromatic hydrocarbon bioremediation and bioelectricity generation.

Microbial Fuel Cell (MFC) is an innovative bio-electrochemical approach which converts biochemical energy inherent in wastewater into electrical energy, thus contributing to circular economy. Five electrogenic bacteria, Kocuria rosea (GTPAS76), two strains of Bacillus circulans (GTPO28 and GTPAS54), and two strains of Corynebacterium vitaeruminis (GTPO38 and GTPO42) were isolated from a common effluent treatment plant (CETP) and were used individually as well as in consortium form to run double chambered "H" type microbial fuel cell. Individually they could produce voltage in the range of 0.4-0.7 V in the MFC systems. Consortium developed using GTPO28, GTPO38, GTPAS54 and GTPAS76 were capable of producing voltage output of 0.8 V with 81.81 % and 64 % COD and BOD reduction, respectively. The EPS production capacity and electricity generation by the isolated bacteria correlated significantly (r = 0.72). Various parameters like, effect of preformed biofilm, length of salt bridge and its reuse, aeration, substrate concentration and external resistance were studied in detail. The study emphasizes on improving the commercialization aspect of MFC with repeated use of salt bridge and improving wastewater treatment potential after optimization of MFC system. Polarization curve and power density trends were studied in optimized MFC. A maximum power density and current density achieved were 18.15 mW/m2 and 370.37 mA/m2, respectively using 5 mM sodium benzoate. This study reports the use of sodium benzoate as a substrate along with reusing of the salt bridge in MFC study with promising results for BOD and COD reduction, proving it to be futuristic technology for bio-based circular ecosystem development.

Uncovering Competitive and Restorative Effects of Macro- and Micronutrients on Sodium Benzoate Biodegradation.

A model aromatic compound, sodium benzoate, is generally used for simulating aromatic pollutants present in textile effluents. Bioremediation of sodium benzoate was studied using the most abundant bacteria, Pseudomonas citronellolis, isolated from the effluent treatment plants of South Gujarat, India. Multiple nutrients constituting the effluent in actual conditions are proposed to have interactive effects on biodegradation which needs to be analyzed strategically for successful field application of developed bioremediation process. Two explicitly different sets of fractional factorial designs were used to investigate the interactive influence of alternative carbon, nitrogen sources, and inorganic micronutrients on sodium benzoate degradation. The process was negatively influenced by the co-existence of other carbon sources and higher concentration of KH2PO4 whereas NH4Cl and MgSO4 exhibited positive effects. Optimized concentrations of NH4Cl, MgSO4, and KH2PO4 were found to be 0.35, 1.056, and 0.3 mg L-1 respectively by central composite designing. The negative effect of high amount of KH2PO4 could be ameliorated by increasing the amount of NH4Cl in the biodegradation milieu indicating the possibility of restoration of the degradation capability for sodium benzoate degradation in the presence of higher phosphate concentration.

Development of fluorescent protein-based biosensing strains: A new tool for the detection of aromatic hydrocarbon pollutants in the environment.

The major sources for release of hydrocarbons into the environment include the effluents generated from chemical processing industries and ports. The introduction of such hazardous compounds into natural water bodies creates considerable disturbances in aquatic life and causes a threat to humans. Thus, it is essential to detect and quantify pollutants at various stages of the wastewater generation and treatment before they reach natural aquatic environments and contaminate them. This study reports the development of "biosensing strains" by cloning hydrocarbon recognizing promoter-operator and a reporter gene in bacterial strains for sensing the presence of pollutants at their lowest possible concentration. So far, various biosensing strains have been constructed with a fused promoter-operator region of the hydrocarbon degrading operons, but most of them use luxAB as a reporter gene. A novel approach in the present study aimed at constructing strains harboring two different fluorescent protein (FP)-based reporter genes for the quantification of multiple pollutants at a time. Two vectors were designed with a fusion of tbuT-gfp and phnR-cfp for the quantification of mono- and poly-aromatic hydrocarbons, respectively. The designed vectors were transformed into E. coli DH5α, and these strains were designated as E. coli DH5α 2296-gfp (containing pPROBE-Tbut-RBS-gfp-npt) and E. coli DH5α 2301-cfp (containing pPROBE-phn-RBS-cfp-npt). Both the developed recombinant strains were capable of successfully detecting mono- and poly-aromatic hydrocarbons in the range of 1-100 µM. The sensing capacity of recombinant strains was successfully validated with actual wastewater samples against available physico-chemical analytical techniques. The development of such recombinant microbial strains indicates the future for online contaminant detection, treatment quality monitoring and protection of aquatic flora and fauna.

Modification of extraction method for community DNA isolation from salt affected compact wasteland soil samples.

To overcome the issue of interferences by salt and compactness in release of bacterial cell required for lysis, method described by Yeates et al. (1998), was optimized for isolation of genomic material (Deoxyribo Nucleic Acid, DNA) from soil microbial community by addition of Al(NH4)SO4. Very low total viable count was observed in the samples tested and hence use of higher amount of soil is required primarily for DNA isolation from wasteland soils. The method proves itself efficient where commercially available bead beating and enzymatic lysis methods could not give isolation of any amount of community genomic DNA due to compact nature and salt concentrations present in soil. •The protocol was found efficient for soil samples with high clay content for microbial community DNA extraction.•Variation in lysis incubation and amount of soil may help with soil samples containing low microbial population.•Addition of Al(NH4)SO4 is crucial step in humic acid removal from extracted DNA samples for soil samples containing high salinity and clay particles.

Functional microbial diversity dynamics in common effluent treatment plants of South Gujarat and hydrocarbon degradation.

Common effluent treatment plants (CETPs) of South Gujarat region, India, process wastewater generated by more than 2500 industries because of the nonfeasibility of processing at the individual industrial unit. This study assessed functional microbial diversity in wastewater samples of CETPs over a geological belt using Ecoplate®, isolation of the most abundant bacteria, and screening for hydrocarbon degradation. The high evenness (EPielou) values (0.9) in almost all samples indicated a highly even community structure. Principal component analysis of carbon source utilization showed a cluster of all inlet samples except E1 and another cluster of all outlet samples; aeration tank community samples were dispersed. In spite of the high richness found in microbial communities, 60 morphologically similar organisms were observed and isolated; 46 out of them were subjected to amplified ribosomal DNA restriction analysis with MboI, HaeIII, and TaqI enzyme, followed by UPGMA clustering. In screening the most abundant bacteria from each cluster, one of the cultures showed a high potential for hydrocarbon degradation and was identified as Pseudomonas citronellolis by 16S rDNA sequencing. Because of its highly adapted inherent nature, this bacterium may help augment the conventional procedure in wastewater treatment and efficiently decrease the organic load.