Environmental pollution has been on the rise in the past few decades owing to increased human activities on energy reservoirs, unsafe agricultural practices and rapid industrialization. Amongst the pollutants that are of environmental and public health concerns due to their toxicities are: heavy metals, nuclear wastes, pesticides, green house gases, and hydrocarbons. Remediation of polluted sites using microbial process (bioremediation) has proven effective and reliable due to its eco-friendly features. Bioremediation can either be carried out ex situ or in situ, depending on several factors, which include but not limited to cost, site characteristics, type and concentration of pollutants. Generally, ex situ techniques apparently are more expensive compared to in situ techniques as a result of additional cost attributable to excavation. However, cost of on-site installation of equipment, and inability to effectively visualize and control the subsurface of polluted sites are of major concerns when carrying out in situ bioremediation. Therefore, choosing appropriate bioremediation technique, which will effectively reduce pollutant concentrations to an innocuous state, is crucial for a successful bioremediation project. Furthermore, the two major approaches to enhance bioremediation are biostimulation and bioaugmentation provided that environmental factors, which determine the success of bioremediation, are maintained at optimal range. This review provides more insight into the two major bioremediation techniques, their principles, advantages, limitations and prospects.

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Bioremediation techniques–classification based on site of application: principles, advantages, limitations and prospects

Environmental pollution from hazardous waste materials, organic pollutants and heavy metals, has adversely affected the natural ecosystem to the detriment of man. These pollutants arise from anthropogenic sources as well as natural disasters such as hurricanes and volcanic eruptions. Toxic metals could accumulate in agricultural soils and get into the food chain, thereby becoming a major threat to food security. Conventional and physical methods are expensive and not effective in areas with low metal toxicity. Bioremediation is therefore an eco-friendly and efficient method of reclaiming environments contaminated with heavy metals by making use of the inherent biological mechanisms of microorganisms and plants to eradicate hazardous contaminants. This review discusses the toxic effects of heavy metal pollution and the mechanisms used by microbes and plants for environmental remediation. It also emphasized the importance of modern biotechnological techniques and approaches in improving the ability of microbial enzymes to effectively degrade heavy metals at a faster rate, highlighting recent advances in microbial bioremediation and phytoremediation for the removal of heavy metals from the environment as well as future prospects and limitations. However, strict adherence to biosafety regulations must be followed in the use of biotechnological methods to ensure safety of the environment.

https://www.mdpi.com/1660-4601/14/12/1504/pdf

Microbial and Plant-Assisted Bioremediation of Heavy Metal Polluted Environments: A Review

A field experiment was conducted in Delaware (USA) to evaluate three crude oil bioremediation techniques. Four treatments were studied: no oil control, oil alone, oil + nutrients, and oil + nutrients + an indigenous inoculum. The microbial populations were monitored by standard MPN techniques, PLFA profile analysis, and 16S rDNA DGGE analysis for species definition. Viable MPN estimates showed high but steadily declining microbial numbers and no significant differences among treatments during the 14-weeks. Regarding the PLFA results, the communities shifted over the 14-week period from being composed primarily of eukaryotes to Gram-negative bacteria. The Gram-negative communities shifted from the exponential to the stationary phase of growth after week 0. All Gram-negative communities showed evidence of environmental stress. The 16S rDNA DGGE profile of all plots revealed eight prominent bands at time zero. The untreated control plots revealed a simple, dynamic dominant population structure throughout the experiment. The original banding pattern disappeared rapidly in all oiled plots, indicating that the dominant species diversity changed and increased substantially over 14 weeks. The nature of this change was altered by nutrient-addition and the addition of the indigenous inoculum.

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Microbial Population Changes During Bioremediation of an Experimental Oil Spill

Biological activity and environmental impact of anionic surfactants

The ability of heavy metals bioaccumulation to cause toxicity in biological systems—human, animals, microorganisms and plants—is an important issue for environmental health and safety. Recent biotechnological approaches for bioremediation include biomineralization (mineral synthesis by living organisms or biomaterials), biosorption (dead microbial and renewable agricultural biomass), phytostabilization (immobilization in plant roots), hyperaccumulation (exceptional metal concentration in plant shoots), dendroremediation (growing trees in polluted soils), biostimulation (stimulating living microbial population), rhizoremediation (plant and microbe), mycoremediation (stimulating living fungi/mycelial ultrafiltration), cyanoremediation (stimulating algal mass for remediation) and genoremediation (stimulating gene for remediation process). The adequate restoration of the environment requires cooperation, integration and assimilation of such biotechnological advances along with traditional and ethical wisdom to unravel the mystery of nature in the emerging field of bioremediation. This review highlights better understanding of the problems associated with the toxicity of heavy metals to the contaminated ecosystems and their viable, sustainable and eco-friendly bioremediation technologies, especially the mechanisms of phytoremediation of heavy metals along with some case studies in India and abroad. However, the challenges (biosafety assessment and genetic pollution) involved in adopting the new initiatives for cleaning-up the heavy metals-contaminated ecosystems from both ecological and greener point of view must not be ignored.

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Biotechnological advances in bioremediation of heavy metals contaminated ecosystems: an overview with special reference to phytoremediation

Microbial growth on and utilization of environmental contaminants as substrates have been studied by many researchers. Most times, substrate utilization results in removal of chemical contaminant, increase in microbial biomass and subsequent biodegradation of the contaminant. These are all aimed at detoxification of the environmental pollutants. Several microbial growth and biodegradation kinetic models have been developed, proposed and used in bioremediation schemes. Some of these models include Monod’s, Andrews, Bungay’s weighted model, general substrate inhibition models (GSIM) and sum kinetic models. Most research on microbial potentials to degrade chemical pollutants has been performed on a laboratory scale. There is a need to extend such studies to pilot scale as well as to full-scale field applications.

	 

	Key words: Microbial growth, substrate utilization, biodegradation, kinetics, detoxification, organic contaminants, models, environmental pollutants.

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Microbial growth and substrate utilization kinetics

Bioremediation of hydrocarbon pollutants is advantageous owing to the cost-effectiveness of the technology and the ubiquity of hydrocarbon-degrading microorganisms in the soil. Soil microbial diversity is affected by hydrocarbon perturbation, thus selective enrichment of hydrocarbon utilizers occurs. Hydrocarbons interact with the soil matrix and soil microorganisms determining the fate of the contaminants relative to their chemical nature and microbial degradative capabilities, respectively. Provided the polluted soil has requisite values for environmental factors that influence microbial activities and there are no inhibitors of microbial metabolism, there is a good chance that there will be a viable and active population of hydrocarbon-utilizing microorganisms in the soil. Microbial methods for monitoring bioremediation of hydrocarbons include chemical, biochemical and microbiological molecular indices that measure rates of microbial activities to show that in the end the target goal of pollutant reduction to a safe and permissible level has been achieved. Enumeration and characterization of hydrocarbon degraders, use of micro titer plate-based most probable number technique, community level physiological profiling, phospholipid fatty acid analysis, 16S rRNA- and other nucleic acid-based molecular fingerprinting techniques, metagenomics, microarray analysis, respirometry and gas chromatography are some of the methods employed in bio-monitoring of hydrocarbon remediation as presented in this review.

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Monitoring of microbial hydrocarbon remediation in the soil.

The bacterial diversity in a tropical soil experimentally polluted with crude oil during a 57 days bioremediation was investigated in five 1 m2 plots using total culturable hydrocarbon utilizing bacteria, heterotrophic bacteria and gas chromatographic analyses. Four out of the five experimental plots received each 4 L of Bonny light crude oil while three treatment plots received 3 kg of NPK, urea fertilizers or poultry droppings with periodic tilling. Two plots, oil-contaminated and pristine served as controls. Bacterial counts increased 200 fold and 2 fold in the NPK treated and poultry-dropping-treated plots respectively, by day 31 post-inoculation. Detectable hydrocarbons in the treatment plots decreased by 84 - 95% and 96 - 99%, 31 and 57 days  post-inoculation, respectively, compared with the petroleum contaminated control. Bacterial strains isolated included Rhodococcus sp., Nocardia sp., Arthrobacter sp., Gordonia sp., Mycobacterium sp., Corynebacterium sp., Bacillus sp., Micrococcus sp., Flavobacterium sp., Pseudomonas sp. and Alcaligenes sp. The overall data suggest an important contribution of Actinobacteria during bioremediation of crude oil-polluted soil.

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Bacterial diversity in a tropical crude oil-polluted soil undergoing bioremediation

Crude oil-polluted marine sediment from Bonny River loading jetty Port Harcourt, Nigeria was treated in seven 2.5 l stirred-tank bioreactors designated BNPK, BNK5, BPD, BNO3, BUNa, BAUT, and BUK over a 56-day period. Five bioreactors were biostimulated with either K2HPO4, NH4NO3, (NH4)2SO4, NPK, urea or poultry droppings while unamended (BUNa) and heat-killed (BAUT) treatments were controls. For each bioreactor, 1 kg (wet weight) sediment amended with 1 l seawater were spiked with 20 ml and 20 mg of crude oil and anthracene which gave a total petroleum hydrocarbons (TPH) range of 106.4–116 ppm on day 0. Polycyclic aromatic hydrocarbons (PAH) in all spiked sediment slurry ranged from 96.6 to 104.4 ppm. TPH in each treatment was ≤14.9 ppm while PAH was ≤6.8 ppm by day 56. Treatment BNO3 recorded highest heterotrophic bacterial count (9.8 × 108 cfu/g) and hydrocarbon utilizers (1.15 × 108 cfu/g). By day 56, the percentages of biodegradation of PAHs, as measured with GC–FID were BNK5 (97.93%), BNPK (98.38%), BUK (98.82%), BUNa (98.13%), BAUT (93.08%), BPD (98.92%), and BNO3 (98.02%). BPD gave the highest degradation rate for PAH. TPH degradation rates were as follows: BNK5 (94.50%), BNPK (94.77%), BUK (94.10%), BUNa (94.77%), BAUT (75.04%), BPD (95.35%), BNO3 (95.54%). Fifty-six hydrocarbon utilizing bacterial isolates obtained were Micrococcus spp. 5 (9.62%), Staphylococcus spp. 3 (5.78%), Pseudomonas spp. 7 (13.46%), Citrobacter sp. 1 (1.92%), Klebsiella sp. 1 (1.92%), Corynebacterium spp. 5 (9.62%), Bacillus spp. 5 (9.62%), Rhodococcus spp. 7 (13.46%), Alcanivorax spp. 7 (13.46%), Alcaligenes sp. 1 (1.92%), Serratia spp. 2 (3.85%), Arthrobacter spp. 7 (13.46%), Nocardia spp. 2 (3.85%), Flavobacterium sp. 1 (1.92%), Escherichia sp. 1 (1.92%), Acinetobacter sp. 1 (1.92%), Proteus sp. 1 (1.92%) and unidentified bacteria 10 (17%). These results indicate that the marine sediment investigated is amenable to bioreactor-based bioremediation and that abiotic factors also could contribute to hydrocarbon attenuation as recorded in the heat-killed (BAUT) control.

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Gideon Chijioke Okpokwasili

Papers

Bioremediation techniques–classification based on site of application: principles, advantages, limitations and prospects

Microbial growth and substrate utilization kinetics

Monitoring of microbial hydrocarbon remediation in the soil.

Bacterial diversity in a tropical crude oil-polluted soil undergoing bioremediation

Bioreactor-based bioremediation of hydrocarbon-polluted Niger Delta marine sediment, Nigeria.