Siti Khodijah Chaerun

Interaction between clay minerals and hydrocarbon-utilizing indigenous microorganisms in high concentrations of heavy oil: implications for bioremediation

Abstract This study focused on whether the presence of clay minerals (montmorillonite and kaolinite) in marine or coastal environments contaminated with high concentrations of heavy-oil spills were able to support the growth of hydrocarbon degraders to enable bioremediation. The bacterial growth experiment utilizing ~150 g/l of heavy oil (from the Nakhodka oil spill) was conducted with 1500 mg/l of montmorillonite or kaolinite. Bacterial strain Pseudomonas aeruginosa (isolated from Atake seashore, Ishikawa Prefecture, Japan), capable of degrading heavy oil, was employed in combination with other hydrocarbon degraders inhabiting the heavy oil and seawater (collected from the Sea of Japan). The interactions among microbial cells, clay minerals and heavy oil were studied. Both clays were capable of promoting microbial growth and allowed microorganisms to proliferate (to a greater degree than in a control sample which contained no clay) in an extremely high concentration of heavy oil. Observation by transmission electron microscopy of the clay-oil-cell complexes showed that microbial cells tended to be bound primarily on the edges of the clays. X-ray diffraction analysis showed that the clay-oil and clay-oil-cell complexes involved the adsorption of microbial cells and/or heavy oil on the external surfaces of the clays. How do the interactions among clay minerals, microbial cells and heavy oil contribute to environmental factors influencing the bioremediation process? To our knowledge, there are no previous reports on the use of clay minerals in the bioremediation of the Nakhodka oil spill in combination with biofilm formation.

Montmorillonite mitigates the toxic effect of heavy oil on hydrocarbon-degrading bacterial growth: implications for marine oil spill bioremediation

Abstract The ability of montmorillonite to mitigate the toxic effect of heavy oil from the Nakhodka oil spill, by growth of hydrocarbon-degrading bacteria and enable bioremediation was studied. Montmorillonite enhanced the bacterial growth significantly (P < 0.05) in the main treatment containing heavy oil+bacteria+montmorillonite (OBM), because the specific growth rate (μ) was greater than that in the biotic control treatment containing heavy oil+bacteria (OB). Significant amounts of Si and Al (major constituents of montmorillonite) were not released in the aqueous phase over the ~24-day experiment (P > 0.05). Transmission electron microscopic observation showed that the hydrocarbon-degrading bacterial cells were covered and encrusted with montmorillonite particles. Scanning transmission electron microscopy coupled with energy dispersive X-ray spectroscopy (STEM-EDS) also showed that the surrounding of the bacterial cells was frequently rich in Si but not in Al. Fourier transform infrared (FTIR) spectroscopy indicated that the heavy oilbacterial cell-montmorillonite particle complex retained the composition of both water and heavy oil. X-ray powder diffractrometery (XRD) analysis revealed that heavy oil and heavy oil-bacteria did not change the basal spacing of montmorillonite over a period of 24 days. The enhancement of hydrocarbon-degrading bacterial growth is attributed to montmorillonite likely serving as both bacterial growth-supporting carrier and protective outer layer against high concentrations of heavy oil that inhibit growth. These results shed light on the interactions in oil-bacteria-clay complexes and could potentially be used in marine oil spill bioremediation.

Effects of Several Parameters on Nickel Extraction from Laterite Ore by Direct Bioelaching Using Aspergillus niger and Acid Rock Drainage from Coal Mine as an Organic Substrate

Aspergillus niger is a prominent fungi that has been used for bioleaching of nickel laterite ore and commercial production of citric acid. Series of shake flask bioleaching assays have been conducted to study the effects of ore type, ore particle size distribution, solid percentage, and substrate volume percentages as well as sulphur addition on the nickel extraction from Indonesian laterite ore. Acid rock drainage (ARD) from local coal mine was used as a substrate for carbon and nutrient sources of the fungus. The results show that saprolite ore exhibits a better leaching with fungal metabolic acids than limonite. The highest nickel extraction of saprolite ore was 43%, which was obtained from the shake flask bioleaching assays with ore particle size distribution of -60+80 mesh, pulp density of 2.5%, and ARD substrate volume percentage of 10%, after 24 days of incubation. For both saprolite and limonite ore samples, coarse particle size ditribution of -60+80 mesh provides a better nickel extraction than the finer ore particle size. The addition of potato dextrose agar and fresh innoculum after 16 incubation days generated a re-increase of nickel extraction, indicating the refreshment of the active fungus. Based on the results, it was found that Aspergilus niger growth can take place with an organic substrate of liquid ARD obtained from coal mining area.

Recovery of Copper from Pyritic Copper Ores Using a Biosurfactant-Producing Mixotrophic Bacterium as Bioflotation Reagent

In an attempt to investigate the use of bacteria and their metabolites as bioflotation reagents for environmentally friendly mineral processing, laboratory cell flotation tests were carried out using copper sulfide ores bearing a high content of pyrite, which were mixed with a biosurfactant-producing mixotrophic bacterium as bioflotation reagents. The interaction of bacterial cells and their metabolic products with the sulfide ores resulted in the alteration of the surface chemistry of both ores and bacterial cells as evidenced by FTIR and SEM-EDS observations as well as surface tension and contact angle measurements. The change in the surface properties of the sulfide ores in turn enabled the bacterium to function as flotation bioregeants in the flotation of copper sulfide ores as a function of bacterial cell concentration, conditioning time, flotation time and pH. Overall, the bacterium and its metabolites as bioreagents yielded flotation recoveries which might be attributed to the multi-function of the bacterium as depressant, collector and frother. Thus, the bacteria tested in this study could potentially be used as flotation bioreagents, providing an alternative to conventional flotation reagents.

Bioleaching of Supergene Porphyry Copper Ores from Sungai Mak Gorontalo of Indonesia by an Iron- and Sulfur-Oxidizing Mixotrophic Bacterium

Environmentally friendly mining technologies have to be developed extensively to avoid the impact of mining activities with respect to environmental concern. One example of such technology is bioleaching which has been developed worldwide and is regarded as an appropriate technology for the extraction of metals from polymetallic ores such as supergene porphyry copper ores, mainly consisting of secondary copper sulfides, including chalcocite (Cu2S), covellite (CuS), or oxide minerals, i.e., cuprite (Cu2O) and tenorite (CuO). The extraction process for this complex ore generally requires high temperature, high pressure and/or high acid concentration. For the economic extraction of valuable copper from such ores, the bioleaching of supergene porphyry copper ore from Sungai Max in Southeast Sulawesi of Indonesia was investigated in shake-flask experiments at room temperature (28 °C) for 14 days using an iron- and sulfur-oxidizing bacterium (Alicyclobacillus sp.) indigenous to an Indonesian mine site. The main mineralogical composition of gangue minerals contained in this ore sample included quartz, muscovite, kaolinite and alunite. The relatively high copper extraction efficiencies were obtained over a 14-day period of the bioleaching experiments as a function of pH, pulp density and NaCl concentration. Therefore, the findings of this study provide the first information of bioleaching process of supergene porphyry copper ores in Indonesia that may highly be useful for developing an economical and environmentally friendly extraction process of such complex ores.

Isolation and phylogenetic characterization of iron-sulfur-oxidizing heterotrophic bacteria indigenous to nickel laterite ores of Sulawesi, Indonesia: Implications for biohydrometallurgy

The main objective of this study was to isolate and phylogenetically identify the indigenous iron-sulfur-oxidizing heterotrophic bacteria capable of bioleaching nickel from laterite mineral ores. The bacteria were isolated from a nickel laterite mine area in South Sulawesi Province, Indonesia. Seven bacterial strains were successfully isolated from laterite mineral ores (strains SKC/S-1 to SKC/S-7) and they were capable of bioleaching of nickel from saprolite and limonite ores. Using EzTaxon-e database, the 16S rRNA gene sequences of the seven bacterial strains were subjected to phylogenetic analysis, resulting in a complete hierarchical classification system, and they were identified as Pseudomonas taiwanensis BCRC 17751 (98.59% similarity), Bacillus subtilis subsp. inaquosorum BGSC 3A28 (99.14% and 99.32% similarities), Paenibacillus pasadenensis SAFN-007 (98.95% and 99.33% similarities), Bacillus methylotrophicus CBMB 205 (99.37% similarity), and Bacillus altitudinis 41KF2b (99.37% similarity). It is n...

Effects of Individual Use, Mixed Culture and Sulfur Addition on the Effectiveness of Nickel Laterite Ore Bioelaching with Penicillium verruculosum and Galactomyces geotrichum

The type of fungi used in fungal bioleaching influence the effectiveness of metals extraction due to the dissimilarity of each microorganism survival in environments with certain metals content. Several metals have prominent toxicities to certain type of fungi, which result in low metals extraction and leaching rate. In the present work, the influences of individual use of Penicillium verruculosum and Galactomyces geotrichum on the nickel extraction from Indonesian laterite ore are compared with the fungis mixed culture. The effect of sulfur addition in the mixed culture of Penicillium verruculosum and Galactomyces geotrichum on the sulfate ion formation was also investigated. Shake flask bioleaching assays have been performed in a medium growth of Indonesian food production waste over the course of 28 days. It was observed that mixed culture of Penicillium verruculosum and Galactomyces geotrichum has a higher effect in nickel extraction compared with the use of individual fungi for both saprolite and limonite ores. For saprolite ore, the fungi species of Galactomyces geotrichum demonstrated a greater nickel extraction than Penicillium verruculosum. In contrast, the extraction of nickel from limonite ore by Penicillium verruculosum was slightly more effective than Galactomyces geotrichum. The addition of elemental sulfur in the mixed culture was found to generate sulfate anion, having a significant increase in its concentration with time and decreasing solution pH within 28-day of incubation. The proportion of sulfur addition must be optimized in order to avoid ore surface passivation with the excess of sulfur. XRD analysis detected the presence of un-oxidized sulfur on surface of the leaching residues along with the biomass.