Krishna K. Kadali

Co-Cultivation of Fungal and Microalgal Cells as an Efficient System for Harvesting Microalgal Cells, Lipid Production and Wastewater Treatment

The challenges which the large scale microalgal industry is facing are associated with the high cost of key operations such as harvesting, nutrient supply and oil extraction. The high-energy input for harvesting makes current commercial microalgal biodiesel production economically unfeasible and can account for up to 50% of the total cost of biofuel production. Co-cultivation of fungal and microalgal cells is getting increasing attention because of high efficiency of bio-flocculation of microalgal cells with no requirement for added chemicals and low energy inputs. Moreover, some fungal and microalgal strains are well known for their exceptional ability to purify wastewater, generating biomass that represents a renewable and sustainable feedstock for biofuel production. We have screened the flocculation efficiency of the filamentous fungus A. fumigatus against 11 microalgae representing freshwater, marine, small (5 µm), large (over 300 µm), heterotrophic, photoautotrophic, motile and non-motile strains. Some of the strains are commercially used for biofuel production. Lipid production and composition were analysed in fungal-algal pellets grown on media containing alternative carbon, nitrogen and phosphorus sources contained in wheat straw and swine wastewater, respectively. Co-cultivation of algae and A. fumigatus cells showed additive and synergistic effects on biomass production, lipid yield and wastewater bioremediation efficiency. Analysis of fungal-algal pellets fatty acids composition suggested that it can be tailored and optimised through co-cultivating different algae and fungi without the need for genetic modification.

Fungal-assisted algal flocculation: application in wastewater treatment and biofuel production

BackgroundThe microalgal-based industries are facing a number of important challenges that in turn affect their economic viability. Arguably the most important of these are associated with the high costs of harvesting and dewatering of the microalgal cells, the costs and sustainability of nutrient supplies and costly methods for large scale oil extraction. Existing harvesting technologies, which can account for up to 50% of the total cost, are not economically feasible because of either requiring too much energy or the addition of chemicals. Fungal-assisted flocculation is currently receiving increased attention because of its high harvesting efficiency. Moreover, some of fungal and microalgal strains are well known for their ability to treat wastewater, generating biomass which represents a renewable and sustainable feedstock for bioenergy production.ResultsWe screened 33 fungal strains, isolated from compost, straws and soil for their lipid content and flocculation efficiencies against representatives of microalgae commercially used for biodiesel production, namely the heterotrophic freshwater microalgae Chlorella protothecoides and the marine microalgae Tetraselmis suecica. Lipid levels and composition were analyzed in fungal-algal pellets grown on media containing alternative carbon, nitrogen and phosphorus sources from wheat straw and swine wastewater, respectively. The biomass of fungal-algal pellets grown on swine wastewater was used as feedstock for the production of value-added chemicals, biogas, bio-solids and liquid petrochemicals through pyrolysis. Co-cultivation of microalgae and filamentous fungus increased total biomass production, lipid yield and wastewater bioremediation efficiency.ConclusionFungal-assisted microalgal flocculation shows significant potential for solving the major challenges facing the commercialization of microalgal biotechnology, namely (i) the efficient and cost-effective harvesting of freshwater and seawater algal strains; (ii) enhancement of total oil production and optimization of its composition; (iii) nutrient supply through recovering of the primary nutrients, nitrogen and phosphates and microelements from wastewater. The biomass generated was thermochemically converted into biogas, bio-solids and a range of liquid petrochemicals including straight-chain C12 to C21 alkanes which can be directly used as a glycerine-free component of biodiesel. Pyrolysis represents an efficient alternative strategy for biofuel production from species with tough cell walls such as fungi and fungal-algal pellets.

A complementary approach to identifying and assessing the remediation potential of hydrocarbonoclastic bacteria

The isolation and assessment of hydrocarbonoclastic bacteria often represents a key strategy in the bioremediation of hydrocarbon-contaminated sites. However the isolation and assessment of such bacteria is often a lengthy and expensive procedure. The aim of this study was to identify potential isolates for use in the remediation of hydrocarbon contaminated sites using a combination of selective isolation plating, the Biolog system and subsequent multivariate analyses. The use of weathered oil as the main C source restricted the number of isolates growing to 5×10(2)CFUg soil(-1). Isolates (n=96) were then assessed individually using Biolog MT2 plates with seven different hydrocarbons (dodecane, tridecane, hexadecane, octadecane, eicosane, naphthalene and phenanthrene). The results indicated that all isolates were able to grow on at least one hydrocarbon from the seven chosen. This confirmed that the isolation media developed was selective in isolating hydrocarbonoclastic bacteria only. Cluster analysis of Biolog data separated the isolates into two discrete clusters with cluster 2 identifying hydrocarbonoclastic bacteria that are effective in degrading a variety of contaminants. Further study on the isolates from cluster 2 was carried out based on their phylogenetic analysis. Phylogenetic analysis of 28 bacterial isolates from cluster 2 based on the 1500bp sequences from 16S rDNA genes using MRBAYES confirmed all isolates as being hydrocarbonoclastic, providing supportive evidence that isolates from cluster 2 have a potential use in bioremediation. This approach could improve both the speed and efficiency of the commercial bioremediation process.

Dual application of duckweed and azolla plants for wastewater treatment and renewable fuels and petrochemicals production

BackgroundShortages in fresh water supplies today affects more than 1 billion people worldwide. Phytoremediation strategies, based on the abilities of aquatic plants to recycle nutrients offer an attractive solution for the bioremediation of water pollution and represents one of the most globally researched issues. The subsequent application of the biomass from the remediation for the production of fuels and petrochemicals offers an ecologically friendly and cost-effective solution for water pollution problems and production of value-added products.ResultsIn this paper, the feasibility of the dual application of duckweed and azolla aquatic plants for wastewater treatment and production of renewable fuels and petrochemicals is explored. The differences in absorption rates of the key wastewater nutrients, ammonium and phosphorus by these aquatic macrophytes were used as the basis for optimization of the composition of wastewater effluents. Analysis of pyrolysis products showed that azolla and algae produce a similar range of bio-oils that contain a large spectrum of petrochemicals including straight-chain C10-C21 alkanes, which can be directly used as diesel fuel supplement, or a glycerin-free component of biodiesel. Pyrolysis of duckweed produces a different range of bio-oil components that can potentially be used for the production of “green” gasoline and diesel fuel using existing techniques, such as catalytic hydrodeoxygenation.ConclusionsDifferences in absorption rates of the key wastewater nutrients, ammonium and phosphorus by different aquatic macrophytes can be used for optimization of composition of wastewater effluents. The generated data suggest that the composition of the petrochemicals can be modified in a targeted fashion, not only by using different species, but also by changing the source plants’ metabolic profile, by exposing them to different abiotic or biotic stresses. This study presents an attractive, ecologically friendly and cost-effective solution for efficient bio-filtration of swine wastewater and petrochemicals production from generated biomass.

Carrier mounted bacterial consortium facilitates oil remediation in the marine environment

Marine oil pollution can result in the persistent presence of weathered oil. Currently, removal of weathered oil is reliant on chemical dispersants and physical removal, causing further disruption. In contrast few studies have examined the potential of an environmentally sustainable method using a hydrocarbon degrading microbial community attached to a carrier. Here, we used a tank mesocosm system (50 l) to follow the degradation of weathered oil (10 g l(-1)) using a bacterial consortium mobilised onto different carrier materials (alginate or shell grit). GCMS analysis demonstrated that the extent of hydrocarbon degradation was dependent upon the carrier material. Augmentation of shell grit with nutrients and exogenous hydrocarbon degraders resulted in 75±14% removal of >C32 hydrocarbons after 12 weeks compared to 20±14% for the alginate carrier. This study demonstrated the effectiveness of a biostimulated and bioaugmented carrier material to degrade marine weathered oil.

Investigating the effectiveness of economically sustainable carrier material complexes for marine oil remediation

The application of bioremediation to marine oil spills is limited due to dilution of either nutrients or hydrocarbonoclastic organisms. This study investigated the effectiveness of three unique natural carrier materials (mussel shells, coir peat and mussel shell/agar complex) which allowed nutrients, hydrocarbonoclastic organisms and oil to be in contact, facilitating remediation. TPH analysis after 30 d showed that mussel shells exhibited the greatest capacity to degrade oil with a 55% reduction (123.3 mg l(-1) from 276 mg l(-1)) followed by mussel shell/agar complex (49%) and coir peat (36%). Both the mussel shells and mussel shell/agar complex carriers were significantly different to the control (P=0.008 and P=0.002, respectively). DGGE based cluster analysis of the seawater microbial community showed groupings based on time rather than carriers. This study demonstrated that inexpensive, accessible waste materials used as carriers of hydrocarbonoclastic bacteria led to significant degradation of hydrocarbon contaminants in seawater.

The assessment of the impact of oil palm and rubber plantations on the biotic and abiotic properties of tropical peat swamp soil in Indonesia

Current intensification of agricultural activities in Indonesia has led to increased use of tropical peat swamp forests for agriculture. Ideally, peat swamp ecosystems should not be disturbed as they provide essential services such as soil erosion control, ecosystem stabilization and moderation of climate and energy fluxes as well as reducing carbon emission and conserving biodiversity. In this study, agricultural land from Giam Siak Kecil–Bukit Batu Biosphere Reserve in Indonesia was evaluated to assess the impact of oil palm (burning and without burning) and rubber (5–10 and >40 years old) plantations on soil properties through comparisons with soils from a natural forest (NF). Substantial changes in the physico-chemical properties of soils from both plantations were observed including significant reductions in soil organic matter (4–18%) and water holding capacity (22–53%), but an increase in bulk density (ρb) (0.08–0.17 g cm−3). A significant increase in bacterial biomass was also observed following conversion of the NF to plantation (p<0.05). However, the oil palm plantation (OPP) (without burning) showed reduced microbial activities and the lowest Shannon diversity values (2.90) compared to other samples. Community-level physiological profiling showed impaired community function only in soils from the OPP but higher CO2 exchange rates in most plantation soils. Soils from the rubber plantation (RP) were less impaired in terms of their natural function and therefore RPs appeared to be more suitable for sustainable agricultural use than OPP.

The application of a carrier-based bioremediation strategy for marine oil spills

The application of recycled marine materials to develop sustainable remediation technologies in marine environment was assessed. The remediation strategy consisted of a shell carrier mounted bacterial consortium composed of hydrocarbonoclastic strains enriched with nutrients (Bioaug SC). Pilot scale studies (5000 l) were used to examine the ability of Bioaug-SC to degrade weathered crude oil (10 g l(-1); initially 315,000±44,000 mg l(-1)) and assess the impacts of the introduction and biodegradation of oil. Total petroleum hydrocarbon mass was effectively reduced by 53.3 (±5.75)% to 147,000 (±21,000) mg l(-1) within 27 weeks. 16S rDNA bacterial community profiling using Denaturant Gradient Gel Electrophoresis revealed that cyanobacteria and Proteobacteria dominated the microbial community. Aquatic toxicity assessment was conducted by ecotoxicity assays using brine shrimp hatchability, Microtox and Phaeodactylum tricornutum. This study revealed the importance of combining ecotoxicity assays with oil chemistry analysis to ensure safe remediation methods are developed.

The microbial removal of toxic waste

The rapid growth of the global chemical industry over the last 35 years has meant that there have been both increased amounts and complexity of toxic waste effluents. Global chemical output increased by 63% in the period from 1996 to 20101; this increase has led to an unprecedented release into the environment of a vast array of chemicals. Bioremediation is now a successful environmental biotechnology used for the remediation of these pollutants, having a number of advantages (for example, cost, environmental friendly means of disposal) over any alternative treatment such as placing in landfill or incineration. Bioremediation offers the opportunity to utilise the natural microbial population to treat the contaminated site, returning the elements making up the contaminants to natural nutrient cycling.