Tanaji More

Extracellular polymeric substances of bacteria and their potential environmental applications

Biopolymers are considered a potential alternative to conventional chemical polymers because of their ease of biodegradability, high efficiency, non-toxicity and non-secondary pollution. Recently, extracellular polymeric substances (EPS, biopolymers produced by the microorganisms) have been recognised by many researchers as a potential flocculent for their applications in various water, wastewater and sludge treatment processes. In this context, literature information on EPS is widely dispersed and is very scarce. Thus, this review marginalizes various studies conducted so far about EPS nature-production-recovery, properties, environmental applications and moreover, critically examines future research needs and advanced application prospective of the EPS. One of the most important aspect of chemical composition and structural details of different moieties of EPS in terms of carbohydrates, proteins, extracellular DNA, lipid and surfactants and humic substances are described. These chemical characteristics of EPS in relation to formation and properties of microbial aggregates as well as degradation of EPS in the matrix (biomass, flocs etc) are analyzed. The important engineering properties (based on structural characteristics) such as adsorption, biodegradability, hydrophilicity/hydrophobicity of EPS matrix are also discussed in details. Different aspects of EPS production process such as bacterial strain maintenance; inoculum and factors affecting EPS production were presented. The important factors affecting EPS production include growth phase, carbon and nitrogen sources and their ratio, role of other nutrients (phosphorus, micronutrients/trace elements, and vitamins), impact of pH, temperature, metals, aerobic versus anaerobic conditions and pure and mixed culture. The production of EPS in high concentration with high productivity is essential due to economic reasons. Therefore, the knowledge about all the aspects of EPS production (listed above) is highly essential to formulate a logical and scientific basis for the research and industrial activities. One of the very important issues in the production/application/biodegradation of EPS is how the EPS is extracted from the matrix or a culture broth. Moreover, EPS matrix available in different forms (crude, loosely bound, tightly bound, slime, capsular and purified) can be used as a bioflocculant material. Several chemical and physical methods for the extraction of EPS (crude form or purified form) from different sources have been analyzed and reported. There is ample information available in the literature about various EPS extraction methods. Flocculability, dewaterability and biosorption ability are the very attractive engineering properties of the EPS matrix. Recent information on important aspects of these properties qualitatively as well as quantitatively has been described. Recent information on the mechanism of flocculation mediated by EPS is presented. Potential role of EPS in sludge dewatering and biosorption phenomenon has been discussed in details. Different factors influencing the EPS ability to flocculate and dewaterability of different suspensions have been included. The factors considered for the discussion are cations, different forms of EPS, concentration of EPS, protein and carbohydrate content of EPS, molecular weight of EPS, pH of the suspension, temperature etc. These factors were selected for the study based upon their role in the flocculation and dewatering mechanism as well the most recent available literature findings on these factors. For example, only recently it has been demonstrated that there is an optimum EPS concentration for sludge flocculation/dewatering. High or low concentration of EPS can lead to destabilization of flocs. Role of EPS in environmental applications such as water treatment, wastewater flocculation and settling, colour removal from wastewater, sludge dewatering, metal removal and recovery, removal of toxic organic compounds, landfill leachate treatment, soil remediation and reclamation has been presented based on the most recent available information. However, data available on environmental application of EPS are very limited. Investigations are required for exploring the potential of field applications of EPS. Finally, the limitations in the knowledge gap are outlined and the research needs as well as future perspectives are highlighted.

Potential use of filamentous fungi for wastewater sludge treatment.

Specific filamentous fungi (FF) have been recognized for sludge treatment and possibly these strains can be utilized for simultaneous bioflocculation, solids and pathogens reduction and, removal and degradation of toxic compounds. Based on current research work and findings, this review provides the state-of-art knowledge on the role of FF (or moulds) in sludge treatment. The proposed theories are presented, critically analyzed and future scope for specific research on utilization of FF for treatment of sludge is recommended.

Bacterial polymer production using pre-treated sludge as raw material and its flocculation and dewatering potential

Sterilization, alkaline-thermal and acid-thermal treatments were applied to different sludge solids concentrations (17.0; 22.4; 29.8; 37.3; 44.8 g/L, respectively) and the pre-treated sludge was used as raw material for Serratia sp.1 to produce extracellular polymeric substances (EPS). After 72 h of fermentation, total EPS of 2.3 and 3.4 g/L were produced in sterilized and alkaline-thermal treated sludge as compared to that of 1.5 g/L in acid-thermal treated sludge. Lower EPS were produced at relatively higher solids concentrations (37.3; 44.8 g/L). Broth, crude forms of capsular and slime EPS were extracted from fermented broths and used as conditioning agents by combining with 150 mg of Ca(2+)/L of kaolin suspensions. Maximum flocculation activity of 79.1% and increased dewatering by 52.2% was achieved using broth and crude capsular EPS, respectively. The results demonstrated that EPS having high flocculating capability could be produced using wastewater sludge as sole raw material.

Biochemical diversity of the bacterial strains and their biopolymer producing capabilities in wastewater sludge

The biochemical characterization of 13 extracellular polymeric substances (EPS) producing bacterial strains were carried out by BIOLOG. The bacterial strains were cultured in sterilized sludge for EPS production. Flocculation and dewatering capabilities of produced EPS (broth, crude slime and capsular) were examined using kaolin suspension combined with calcium (150 mg of Ca(2+)/L of kaolin suspension). BIOLOG revealed that there were 9 Bacillus, 2 Serratia and 2 Yersinia species. Most of these bacterial strains had the capability to utilize wide spectrum of carbon and nitrogen sources. EPS concentration of more than 1g/L was produced by most of the bacterial strains. Concentration of EPS produced by different Bacillus strains was higher than that of Serratia and Yersinia. Broth EPS revealed flocculation activity more than 75% for Bacillus sp.7, Bacillus sp.4 and Bacillus sp.6, respectively. Flocculation activity higher than 75% was attained using very low concentrations of broth EPS (1.12-2.70 mg EPS/g SS).

Biopolymer Production Kinetics of Mixed Culture Using Wastewater Sludge as a Raw Material and the Effect of Different Cations on Biopolymer Applications in Water and Wastewater Treatment.

Thirteen extracellular polymeric substances (EPS) producing bacterial strains were cultivated (as mixed culture) in the sterilized sludge (suspended solids of 25 g/L) and the batch fermentation was carried out. Mixed culture revealed a high specific growth rate of 0.35/hr. The EPS production rate was higher up to 24 hours, which gradually decreased with further incubation. The kinetic estimates demonstrated growth-associated EPS production. Broth EPS revealed higher flocculation activity when combined with different cations (Ca(2+), Mg(2+), Fe(3+), and Al(3+)) in river water (≥90%), municipal wastewater (≥90%), and brewery wastewater (≥80%), respectively. A low dose (5 to 40 mg/L) of trivalent cations was required to achieve higher flocculation compared to the divalent cations (50 to 250 mg/L). Flocculation performance of EPS was comparable to Magnafloc-155 (chemical polymer) and, hence, it could be used as a flocculant.

Extracellular polymeric substances production kinetics of 13 sludge isolates using wastewater sludge as raw material and its flocculation potential

The kinetics of batch fermentation of 13 extracellular polymeric substances (EPS) producing bacterial strains (9 Bacillus, 2 Serratia and 2 Yersinia) were carried out using sterilized sludge as a raw material. The most of Bacillus (µmax: 0.11–0.27 h−1), Serratia (µmax: 0.23–0.27 h−1) and Yersinia (µmax: 0.18–0.19 h−1) strains had capability to grow and produce EPS (1.36–2.12 g/L) in the sterilized sludge. In general, EPS production was mixed growth associated for all the bacterial strains cultivated independently. Bacillus sp. 7, Serratia sp. 2 and Yersinia sp. 2 produced higher concentration (1.95–2.12 g/L) of EPS than the other remaining bacterial strains. Protein and carbohydrate contents of EPS remained constant during fermentation. Broth EPS (B-EPS) exhibited high kaolin flocculation activity (≥75%) in most of the cases except Bacillus sp. 1, Bacillus sp. 5 and Bacillus sp. 9, respectively. In general, high flocculation activities (FAs) (≥75%), were attained using 1.31–1.70 mg B-EPS/g kaolin, 0.45–0.97 mg protein/g kaolin and 0.11–0.21 mg carbohydrates/g kaolin. The study suggests that further systematic exploration is required for optimizing the process of EPS production. EPS produced in the sludge can potentially be used for different water and wastewater treatments.

Biopolymers Production by Mixed Culture and Their Applications in Water and Wastewater Treatment.

Thirteen extracellular polymeric substances (EPS) producing bacterial strains were cultivated (as pure/mixed culture) in sterilized sludge (suspended solids: 25 g/L). The mixed culture produced higher concentrations of EPS (4.9 g/L) as compared to that of the pure culture (2.7-3.7 g/L). The harvested EPS were examined for their flocculation performance (turbidity removal and dewatering) in jar tests using kaolin suspensions with Ca2+. Broth (B-EPS) revealed high kaolin flocculating activity (91.2%) at very low concentrations (0.8 mg B-EPS/g kaolin) and it was comparable to the chemical polymer, Magnafloc-155 (90.4% at 0.2 mg/g kaolin). B-EPS also exhibited very good flocculation performance (turbidity removal %) in river water (93.5%), municipal wastewater (91.7%) and brewery wastewater (81.8%). The study revealed that the mixed culture consortium could be used for the production of highly efficient flocculants.

Carbon burial and enhanced soil carbon trapping

Sponsored by the Carbon Capture and Storage Task Committee of the Technical Committee on Hazardous, Toxic, and Radioactive Waste Engineering of the Environmental Council of EWRI.