Optimising Wastewater Treatment: Acinetobacter sp. IrC1 as a potential multi-resistant bacterium for copper accumulation and dyes decolourisation.

Improper disposal of waste containing copper and dye is an environmental issue that must be resolved immediately due to its harmful, non-degradable and toxic properties. Bioremediation efficiency can improve by cultivating copper and dye multi-resistant bacteria to remove various pollutant types simultaneously. This study aims at establishing the multi-resistance of Acinetobacter sp. IrC1 to copper and dyes. The effects of copper concentration on growth were determined using a spectrophotometer, while accumulation was analysed using an atomic absorption spectrophotometer. Bacteria-mediated dye decolourisation dyes were observed based on clear zone formation around bacterial colonies, while decolourisation percentage was calculated using a spectrophotometer. Results demonstrate that Acinetobacter sp. IrC1 resisted up to 8 mM CuSO4 and accumulated up to 292.93 mg/g dry weight of copper cells. Acinetobacter sp. IrC1 isolates were also resistant to 500 ppm Methylene Blue, Malachite Green, Congo Red, Mordant Orange, Reactive Black, Direct Yellow, Reactive Orange, Remazol, Wantex Red and Wantex Yellow dye, successfully removing up to 68.35% and 79.50% Methylene Blue and Basic Fuchsine in a medium containing 3 mM CuSO4, respectively. Further investigations are required to analyse the genetic composition of multi-resistant bacteria to optimise the effectiveness of indigenous bacterial isolates as bioremediation agents.

Two strains of airborne Nocardiopsis alba producing different volatile organic compounds (VOCs) as biofungicide for Ganoderma boninense.

Nocardiopsis are actinobacteria which produce active compounds, such as antifungals and volatile compounds. Ganoderma boninense is a pathogenic and aggressive fungus that decreases palm oil yield during production. In this study, we isolated two strains of Nocardia (GME01 and GME22) from airborne contaminants on the actinobacteria culture collection in the laboratory. The aim of this study is to identify two strains of Nocardiopsis and to obtain the antifungal potency of volatile organic compounds (VOCs) against G. boninese. We characterized the morphology using Scanning Electrone Microscope (SEM), molecular properties and whole-cell protein spectra using Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS), antifungal assay on G. boninense and VOCs analysis of Nocardia using solid phase micro extraction/gas chromatography (SPME/GC). The two Nocardiopsis strains had the similar characteristic such as white aerial mycelium and spores, aerobic, grow well on ISP-2, TSA and NA medium without diffusible pigment and had the highest similarity with Nocardiopsis alba DSM 43377 (99.63% and 99.55% similarity for GME01 and GME22, respectively), Different morphological feature was found in aerial mycelium and spores. GME22 has a clearly fragmented mycelium whereas GME01 has none. Other features also showed different on the whole-cell protein spectra, antifungal activity and VOCs profiles. Antifungal activity assay on G. boninense showed that N. alba GME22 has higher antifungal activity than GME01 related with the VOCs abundance in two strains. Almost 38.3% (18 VOCs) of N. alba GME22 and 25.5% (12 VOCs) of N. alba GME01 were found specifically in each strain, and 36.2% (the 17 same VOCs) produced by both. The known volatile antifungal compounds S-methyl ethanethioate, 1,2-dimethyldisulfane, acetic acid, 2-methyl propanoic acid, 3-methyl-butanoic acid, nonan-2-one, undecan-2-one and 2-isopropyl-5-methylcyclohexan-1-ol only produced by N. alba GME22 and 1,3-dimethyltrisulfane only produced by N. alba GME01. A total of two known antifungal compounds 1,2-dimethyldisulfane and 6-methylheptan-2-one were produced by both N. alba. The abundance of antifungal VOCs produced by these bacteria is potentially to be used as biocontrol agent for pathogenic fungi in plants.

Potential application of thermophilic bacterium Aeribacillus pallidus MRP280 for lead removal from aqueous solution.

Bacteria used for application of lead (Pb) removal is usually kept under suboptimal growth conditions. Certain application of Pb removal may be carried out under different condition, such as under aqueous and high temperature conditions. It is, therefore, of interest to examine the Pb removal capacity of the bacteria under adverse environmental conditions. In the present study, Aeribacillus pallidus MRP280, a lead-tolerant thermophilic bacterium was used as an absorbent for the removal of Pb from aqueous solution. The Pb removal and uptake capacity of living and non-living bacterial cells of A. pallidus MRP280 was investigated in 100 mg/L Pb solution. The optimum condition was examined based on several analytical parameters, including temperature, pH, contact time, and cell density. Biosorbent analysis and characterization was carried out using Fourier Transform Infrared (FT-IR) spectroscopy, Scanning Electron Microscope (SEM)-Energy Dispersive X-ray (EDX), and Transmission Electron Microscope (TEM). The results showed that the maximum Pb removal of 96.78 ± 0.19% and 88.64 ± 0.60% were obtained using living and non-living biomass, respectively at 55 °C, pH 6, OD6000.5 for 100 min. Meanwhile, the maximum uptake capacity of 86.47 ± 1.32 mg/g and 85.31 ± 1.37 mg/g by living and non-living cells was reached at 55 °C, pH 6, OD6000.25 for 60 min. Moreover, Pb removing activity was facilitated by the biosorption and bioaccumulation process. Overall, it is shown that A. pallidus MRP280 is effective when applied as biosorbent in removing Pb from contaminated wastewater at high temperatures.

Optimizing Bioremediation: Elucidating Copper Accumulation Mechanisms of Acinetobacter sp. IrC2 Isolated From an Industrial Waste Treatment Center.

Acinetobacter sp. IrC2 is a copper-resistant bacterium isolated from an industrial waste treatment center in Rungkut, Surabaya. Copper-resistant bacteria are known to accumulate copper inside the cells as a mechanism to adapt to a copper-contaminated environment. Periplasmic and membrane proteins CopA and CopB have been known to incorporate copper as a mechanism of copper resistance. In the present study, protein profile changes in Acinetobacter sp. IrC2 following exposure to copper stress were analyzed to elucidate the copper resistance mechanism. Bacteria were grown in a Luria Bertani agar medium with and without CuSO4 supplementation. Intracellular copper ion accumulation was quantified using atomic absorption spectrophotometry. Changes in protein profile were assessed using sodium dodecyl sulfate polyacrylamide gel electrophoresis. The results showed that 6 mM CuSO4 was toxic for Acinetobacter sp. IrC2, and as a response to this copper-stress condition, the lag phase was prolonged to 18 h. It was also found that the bacteria accumulated copper to a level of 508.01 mg/g of cells' dry weight, marked by a change in colony color to green. The protein profile under copper stress was altered as evidenced by the appearance of five specific protein bands with molecular weights of 68.0, 60.5, 38.5, 24.0, and 20.5 kDa, suggesting the presence of CopA, multicopper oxidase (MCO), CopB, universal stress protein (Usp), and superoxide dismutase (SOD) and/or DNA-binding protein from starved cells, respectively. We proposed that the mechanism of bacterial resistance to copper involves CopA and CopB membrane proteins in binding Cu ions in the periplasm and excreting excess Cu ions as well as involving enzymes that play a role in the detoxification process, namely, SOD, MCO, and Usp to avoid cell damage under copper stress.