Suresh R. Subashchandrabose

Consortia of cyanobacteria/microalgae and bacteria: Biotechnological potential

Microbial metabolites are of huge biotechnological potential and their production can be coupled with detoxification of environmental pollutants and wastewater treatment mediated by the versatile microorganisms. The consortia of cyanobacteria/microalgae and bacteria can be efficient in detoxification of organic and inorganic pollutants, and removal of nutrients from wastewaters, compared to the individual microorganisms. Cyanobacterial/algal photosynthesis provides oxygen, a key electron acceptor to the pollutant-degrading heterotrophic bacteria. In turn, bacteria support photoautotrophic growth of the partners by providing carbon dioxide and other stimulatory means. Competition for resources and cooperation for pollutant abatement between these two guilds of microorganisms will determine the success of consortium engineering while harnessing the biotechnological potential of the partners. Relative to the introduction of gene(s) in a single organism wherein the genes depend on the regulatory- and metabolic network for proper expression, microbial consortium engineering is easier and achievable. The currently available biotechnological tools such as metabolic profiling and functional genomics can aid in the consortium engineering. The present review examines the current status of research on the consortia, and emphasizes the construction of consortia with desired partners to serve a dual mission of pollutant removal and commercial production of microbial metabolites.

Mixotrophic cyanobacteria and microalgae as distinctive biological agents for organic pollutant degradation

Millions of natural and synthetic organic chemical substances are present in both soil and aquatic environments. Toxicity and/or persistence determine the polluting principle of these substances. The biological responses to these pollutants include accumulation and degradation. The responses of environments with organic pollutants are perceptible from the dwindling degradative abilities of microorganisms. Among different biological members, cyanobacteria and microalgae are highly adaptive through many eons, and can grow autotrophically, heterotrophically or mixotrophically. Mixotrophy in cyanobacteria and microalgae can provide many competitive advantages over bacteria and fungi in degrading organic pollutants. Laboratory culturing of strict phototrophic algae has limited the realization of their potential as bioremediation agents. In the natural assemblages, mixotrophic algae can contribute to sequestration of carbon, which is otherwise emitted as carbon dioxide to the atmosphere under heterotrophic conditions by other organisms. Molecular methods and metabolic and genomic information will help not only in identification and selection of mixotrophic species of cyanobacteria and microalgae with capabilities to degrade organic pollutants but also in monitoring the efficiency of remediation efforts under the field conditions. These organisms are relatively easier for genetic engineering with desirable traits. This review presents a new premise from the literature that mixotrophic algae and cyanobacteria are distinctive bioremediation agents with capabilities to sequester carbon in the environment.

p-NITROPHENOL TOXICITY TO AND ITS REMOVAL BY THREE SELECT SOIL ISOLATES OF MICROALGAE: THE ROLE OF ANTIOXIDANTS

The nontarget effects, in terms of biochemical changes induced by p-nitrophenol (PNP) in three soil microalgae, Chlorella sp., Chlorococcum sp., and Heterochlamydomonas sp., and the PNP removal efficiency of these isolates, were determined. On exposure to 20 mg L(-1) PNP, Chlorella sp. showed greater activity of peroxidase, superoxide dismutase, and glutathione reductase as well as high contents of proline and carotenoids. While Heterochlamydomonas sp. exhibited higher levels of catalase and protein, Chlorococcum sp. produced greater amounts of malondialdehyde, a measure of lipid peroxidation, in the presence of PNP. Chlorella sp. tolerated PNP by producing large quantities of antioxidants coupled with less lipid peroxidation, while Chlorococcum sp. was susceptible, as evidenced by low antioxidant production and high lipid peroxidation. During 7-d exposure, Chlorella sp., Heterochlamydomonas sp., and Chlorococcum sp. were able to remove 39, 18, and 4% of 20 mg L(-1) PNP, respectively. The present results indicate that proline, carotenoids and malondialdehyde are the potential biomarkers for assessing PNP toxicity toward microalgae, and their response could be considered for differentiating tolerant and susceptible strains. Moreover, there is a clear correlation between PNP removal and antioxidant synthesis in microalgae on exposure to the pollutant.

Cultivation of Chlorella on brewery wastewater and nano-particle biosynthesis by its biomass

This study investigated an integrated and sustainable approach for iron nanoparticles synthesis using Chlorella sp. MM3 biomass produced from the remediation of brewery wastewater. The algal growth characteristics, biomass production, nutrient removal, and nanoparticle synthesis including its characterisation were studied to prove the above approach. The growth curve of Chlorella depicted lag and exponential phase characteristics during the first 4days in a brewery wastewater collected from a single batch of brewing process (single water sample) indicating the growth of algae in brewery wastewater. The pollutants such as total nitrogen, total phosphorus and total organic carbon in single water sample were completely utilised by Chlorella for its growth. The X-ray photoelectron spectroscopy spectra showed peaks at 706.56eV, 727.02eV, 289.84eV and 535.73eV which corresponded to the zero-valent iron, iron oxides, carbon and oxygen respectively, confirming the formation of iron nanoparticle capped with algal biomolecules. Scanning electron microscopy and particle size analysis confirmed the presence of spherical shaped iron nanoparticles of size ranging from 5 to 50nm. To our knowledge, this is the first report on nanoparticle synthesis using the biomass generated from phycoremediation of brewery wastewater.

Interaction effects of polycyclic aromatic hydrocarbons and heavy metals on a soil microalga, Chlorococcum sp. MM11

Environmental risk assessment of sites contaminated with chemicals needs to also consider mixtures of chemicals as these toxicants act more differently in a mixture than when they occur alone. In this study, we describe, for the first time, the use of a full factorial design experiment to evaluate the toxicity of a quaternary mixture comprising two polycyclic aromatic hydrocarbons (PAHs; benzo[a]pyrene (BaP) and phenanthrene (Phe)) and two heavy metals (cadmium (Cd) and lead (Pb)) toward a soil microalga, Chlorococcum sp. MM11. Biomass, in terms of cell number, and proline accumulation were used to evaluate toxicity responses. Factorial analysis of the data revealed statistically significant interaction effects between the mixtures of toxicants on 96-h biomass endpoint, while no significant interaction effects were observed on proline accumulation in the microalga. A comparison of the data on the toxicity of individual chemicals and those of the factorial main effect analysis clearly showed that Cd is more toxic to the alga, followed by BaP, Pb, and Phe. There was a substantial heavy metal accumulation and PAH degradation by the strain MM11 at EC10 and EC50 of the chemical mixtures.

Microbes from mined sites: Harnessing their potential for reclamation of derelict mine sites

Derelict mines pose potential risks to environmental health. Several factors such as soil structure, organic matter, and nutrient content are the greatly affected qualities in mined soils. Soil microbial communities are an important element for successful reclamation because of their major role in nutrient cycling, plant establishment, geochemical transformations, and soil formation. Yet, microorganisms generally remain an undervalued asset in mined sites. The microbial diversity in derelict mine sites consists of diverse species belonging to four key phyla: Proteobacteria, Acidobacteria, Firmicutes, and Bacteroidetes. The activity of plant symbiotic microorganisms including root-colonizing rhizobacteria and ectomycorrhizal fungi of existing vegetation in the mined sites is very high since most of these microbes are extremophiles. This review outlines the importance of microorganisms to soil health and the rehabilitation of derelict mines and how microbial activity and diversity can be exploited to better plan the soil rehabilitation. Besides highlighting the major breakthroughs in the application of microorganisms for mined site reclamation, we provide a critical view on plant-microbiome interactions to improve revegetation at the mined sites. Also, the need has been emphasized for deciphering the molecular mechanisms of adaptation and resistance of rhizosphere and non-rhizosphere microbes in abandoned mine sites, understanding their role in remediation, and subsequent harnessing of their potential to pave the way in future rehabilitation strategies for mined sites.

Diplosphaera sp. MM1 – A microalga with phycoremediation and biomethane potential

This study evaluated the potential of a microalga Diplosphaera sp. MM1 for its ability to generate energy through biomass production from wastewater remediation. 33% dairy wastewater and 50% winery wastewater demonstrated as promising alternative media for cultivating Diplosphaera sp. MM1 biomass. Interestingly, the alga cultivated in 50% winery wastewater with limited nitrogen produced the highest lipid content (43.07% total solid) and the lowest carbohydrate content (9.35% TS). On the contrary, the lowest lipid content (16.98% TS) and the highest carbohydrate content (29.39% TS) were exhibited by the alga cultivated in 33% dairy wastewater. The results from anaerobic digestion processes in terms of biochemical methane potential of the alga cultivated in BG-11 medium, 33% dairy wastewater and 50% winery wastewater were 197.39, 129.75 and 218.51NmLg(-1)VS, respectively. Further, this study demonstrates the potential of winery wastewater as a candidate to increase the lipid content of algae and enhance biofuel production of algal biomass.

Assessment of bioavailability of heavy metal pollutants using soil isolates of Chlorella sp.

Biotests conducted with plants are presently used to estimate metal bioavailability in contaminated soils. But when plants are grown in soils, especially the plants with fine roots, root collection is easily biased and tedious. Indeed, at harvest, small amounts of soil can adhere to roots, resulting in overestimation of root metal content, and the finest roots are often discarded from the analysis because of their difficult and almost impossible recovery. This report presents a novel method for assessing the bioavailability of heavy metals in soils using microalgae. Two species of green unicellular microalgae were isolated from two highly contaminated soils and identified by phylogenetic and molecular evolutionary analyses as Chlorella sp. RBM and Chlorella sp. RHM. These two cultures were used to determine the metal uptake from metal-contaminated soils of South Australia as a novel, cost-effective, simple and rapid method for assessing the bioavailability of heavy metals in soils. The suggested method is an attempt to achieve a realistic estimate of bioavailability which overcomes the inherent drawback of root metal contamination in the bioavailability indices so far reported.

Rhodococcus wratislaviensis strain 9: An efficient p-nitrophenol degrader with a great potential for bioremediation

A Gram-positive bacterium, Rhodococcus wratislaviensis strain 9, was isolated from groundwater contaminated with nitrophenolics and trichloroethene following enrichment culture technique. The cells of strain 9 grown on LB broth (uninduced) degraded 720 μM p-nitrophenol (PNP) within 12 h, and utilized as a source of carbon and energy. Orthogonal experimental design analysis to determine optimal conditions for biodegradation of PNP showed that pH had a significant positive effect (P ≤ .05) on bacterial degradation of PNP, while glucose, di- and tri-nitrophenols exhibited significant negative effect. Cell-free extracts obtained from PNP-grown culture that contained 20 μg mL-1 protein degraded 90% of 720 μM PNP within 5 h of incubation. Two-dimensional protein analysis revealed differential expression of the oxygenase component of PNP monooxygenase and an elongation factor Tu in PNP-grown cells, but not in those grown on glucose. The strain 9 remediated laboratory wastewater containing 900 μM PNP efficiently within 14 h, indicating its great potential in bioremediation of PNP-contaminated waters.

Microalgae–bacteria biofilms: a sustainable synergistic approach in remediation of acid mine drainage

Microalgae and bacteria offer a huge potential in delving interest to study and explore various mechanisms under extreme environments. Acid mine drainage (AMD) is one such environment which is extremely acidic containing copious amounts of heavy metals and poses a major threat to the ecosystem. Despite its extreme conditions, AMD is the habitat for several microbes and their activities. The use of various chemicals in prevention of AMD formation and conventional treatment in a larger scale is not feasible under different geological conditions. It implies that microbe-mediated approach is a viable and sustainable alternative technology for AMD remediation. Microalgae in biofilms play a pivotal role in such bioremediation as they maintain mutualism with heterotrophic bacteria. Synergistic approach of using microalgae–bacteria biofilms provides supportive metabolites from algal biomass for growth of bacteria and mediates remediation of AMD. However, by virtue of their physiology and capabilities of metal removal, non-acidophilic microalgae can be acclimated for use in AMD remediation. A combination of selective acidophilic and non-acidophilic microalgae together with bacteria, all in the form of biofilms, may be very effective for bioremediation of metal-contaminated waters. The present review critically examines the nature of mutualistic interactions established between microalgae and bacteria in biofilms and their role in removal of metals from AMDs, and consequent biomass production for the yield of biofuel. Integration of microalgal–bacterial consortia in fuel cells would be an attractive emerging approach of microbial biotechnology for AMD remediation.