Enterobacter tabaci 4M9 (CCB-MBL 5004) was reported to have plant growth-promoting and heavy metal tolerance traits. It was able to tolerate more than 300 mg/L Cd, 600 mg/L As, and 500 mg/L Pb and still maintained the ability to produce plant growth-promoting substances under metal stress conditions. To explore the genetic basis of these beneficial traits, the complete genome sequencing of 4M9 was carried out using Pacific Bioscience (PacBio) sequencing technology. The complete genome consisted of one chromosome of 4,654,430 bp with a GC content of 54.6% and one plasmid of 51,135 bp with a GC content of 49.4%. Genome annotation revealed several genes involved in plant growth-promoting traits, including the production of siderophore, indole acetic acid, and 1-aminocyclopropane-1-carboxylate deaminase; solubilization of phosphate and potassium; and nitrogen metabolism. Similarly, genes involved in heavy metals (As, Co, Zn, Cu, Mn, Se, Cd, and Fe) tolerance were detected. These support its potential as a heavy metal-tolerant plant growth-promoting bacterium and a good genetic resource that can be employed to improve phytoremediation efficiency of heavy metal-contaminated soil via biotechnological techniques. This, to the best of our knowledge, is the first report on the complete genome sequence of heavy metal-tolerant plant growth-promoting E. tabaci.
Polyethylene is a widely used packaging material, but its non-biodegradable nature can lead to waste
disposal problems. This increases the concern in research and development of biodegradable plastics from natural resource as alternatives to petroleum-derived plastics. In this study, biodegradable plastic composites were prepared by blending thermoplastic starch with natural rubber in the present of glycerol as plasticizer. Local sago starch was cast with 0.5 to 10% of natural rubber to prepare the bioplastic. The products were characterized by differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), water absorption test, biodegradable test, hydrolysis test, and mechanical analysis. Meanwhile, composite with natural rubber latex was increased from 0.5 to 10% showing that the melting temperature is in the range of 120 to 150˚C, but with no significant difference. The water absorption characteristics, biodegradability, and tensile strength decreased by 11.21%, 30.18%, and 20.733 MPa, respectively. However, the elongation at break was increased from 26.67 to 503.3%. The findings of this study showed that sago starch has a great potential in bioplastic production with good miscibility and compatibility.
Composting can potentially remove organic pollutants in sewage sludge. When estimating pollutant removal efficiency, knowledge of estimate uncertainty is important for understanding estimate reliability. In this study the uncertainty (coefficient of variation, CV) in pollutant degradation rate (K1) and relative concentration at 35days of composting (C35/C0) was evaluated. This was done based on recently presented pollutant concentration data, measured under full-scale composting conditions using two different sampling methods for a range of organic pollutants commonly found in sewage sludge. Non-parametric statistical procedures were used to estimate CV values for K1 and C35/C0 for individual pollutants. These were then used to compare the two sampling methods with respect to CV and to determine confidence intervals for average CV. Results showed that sampling method is crucial for reducing uncertainty. The results further indicated that it is possible to achieve CV values for both K1 and C35/C0 of about 15%.
The recalcitrant landfill leachate was anaerobically digested at various mixing ratios with labile synthetic wastewater to evaluate the degradation properties of recalcitrant wastewater. The proportion of leachate to the digestion system was increased in three equal steps, starting from 0% to 100%, and later decreased back to 0% with the same steps. The chemical oxygen demand (COD) for organic carbon and other components were calculated by analyzing the COD and dissolved organic carbon (DOC), and the removal efficiencies of COD carbon and COD others were evaluated separately. The degradation properties of COD carbon and COD others shifted owing to changing of substrate degradability, and the removal efficiencies of COD carbon and COD others were improved after supplying 100% recalcitrant wastewater. The UV absorptive property and total organic carbon (TOC) of each molecular size using high performance liquid chromatography (HPLC)-size exclusion chromatography (SEC) with UVA and TOC detectors were also investigated, and the degradability of different molecular sizes was determined. Although the SEC system detected extracellular polymeric substances (EPS), which are produced by microbes in stressful environments, during early stages of the experiment, EPS were not detected after feeding 100% recalcitrant wastewater. These results suggest that the microbes had acclimatized to the recalcitrant wastewater degradation. The high removal rates of both COD carbon and COD others were sustained when the proportion of labile wastewater in the substrate was 33%, indicating that the effective removal of recalcitrant COD might be controlled by changing the substrate's degradability.
Phytoremediation is an environmentally friendly and sustainable alternative for treatment of nitrogen-enriched wastewaters. In this study, Ta-khian (Hopea odorata) and Lagos mahogany (Khaya ivorensis), two tropical timber plants, were investigated for their performances in treatment of urea manufacturing factory effluent with high nitrogen (N) content. Plant seedlings received four concentrations of N (190, 240, 290 and 340 mg/L N) in laboratory-scale constructed wetlands every 4 days for a duration of 8 weeks. The solution volumes supplied to each container, amount of N recovered by plants and plant growth characteristics were measured throughout the experiment. Results showed that Ta-khian plants were highly effective at reducing N concentration and volume of water. A maximum of 63.05% N recovery was obtained by Ta-khian plants grown in 290 mg/L N, which was assimilated in the chlorophyll molecule structure and shoot biomass. Significant positive correlations have been shown between N recovery percentages and plant growth parameters. Ta-Khian plants can be applied as suitable phytoremediators for mitigating N pollution in water sources.
Highly photocatalytically active copper chromite nanostructured material were prepared via a novel simple hydrothermal reaction between [Cu(en)2(H2O)2]Cl2 and [Cr(en)3]Cl3.3H2O at low temperature, without adding any pH regulator or external capping agent. The as-synthesized nanostructured copper chromite was analyzed by transmission electron microscopy (TEM), UV-vis diffuse reflectance spectroscopy, energy dispersive X-ray microanalysis (EDX), scanning electron microscopy (SEM), X-ray diffraction (XRD) and Fourier transform infrared (FT-IR) spectroscopy. Results of the morphological investigation of the as-synthesized products illustrate that the shape and size of the copper chromite depended on the surfactant sort, reaction duration and temperature. Moreover, the photocatalytic behavior of as-obtained copper chromite was evaluated by photodegradation of acid blue 92 (anionic dye) as water pollutant.
Previously, a unique co-compost produced by composting empty fruit bunch with anaerobic sludge from palm oil mill effluent, which contributed to establishing a zero-emission industry in Malaysia. Little was known about the bacterial functions during the composting process and fertilization capacity of this co-compost. We isolated 100 strains from the co-compost on 7 types of enumeration media and screened 25 strains using in vitro tests for 12 traits, grouping them according to three functions: plant growth promoting (fixation of nitrogen; solubilization of phosphorus, potassium, and silicate; production of 3-indoleacetic acid, ammonia, and siderophore), biocontrolling (production of chitinase and anti-Ganoderma activity), and composting (degradation of lignin, xylan, and cellulose). Using 16S rRNA gene sequence analysis, 25 strains with strong or multi-functional traits were found belong to the genera Bacillus, Paenibacillus, Citrobacter, Enterobacter, and Kosakonia. Furthermore, several strains of Citrobacter sedlakii exhibited a plant growth-stimulation in vivo komatsuna plant cultivation test. In addition, we isolated several multifunctional strains; Bacillus tequilensis CE4 (biocontrolling and composting), Enterobacter cloacae subsp. dissolvens B3 (plant growth promoting and biocontrolling), and C. sedlakii CESi7 (plant growth promoting and composting). Some bacteria in the co-compost play significant roles during the composting process and plant cultivation after fertilization, and some multifunctional strains have potential for use in accelerating the biodegradation of lignocellulosic biomass, protecting against Ganoderma boninense infection, and increasing the yield of palm oil.
The indiscriminate released of heavy metals and xenobiotics into soils and aquatic bodies
severely alter soil organisms and the ecosystem. The isolation of xenobiotics degrading
microorganisms is cost-effective and naturally pleasant approach. Lately, the toxicological effect
of molybdenum to the spermatogenesis of several organisms has been record. This present study
is aimed at the isolation and characterization of a bacterium capable of converting molybdenum
to the colloidal molybdenum blue. Bacteria characterization was performed in a microplate
format using resting cells. Thus, the reduction process can be employed as a device for
molybdenum bioremediation. The results of the study revealed an optimum reduction at pH
between 6.0 and 6.3 and temperatures of between 25 and 40 oC. Similarly, it was also observed
that a phosphate concentration not greater than 5.0 mM and a sodium molybdate concentration
at 20 mM was required for reduction. Glucose was observed as the best carbon source to support
reduction. Following the scanning of molybdenum blue, it revealed an absorption spectrum
indicating the characteristics of molybdenum blue as a reduced phosphomolybdate. Molybdenum
reduction is inhibited by heavy metals like silver, lead, arsenic and mercury. Furthermore, the
ability of the bacterium (Pseudomonas sp. strain Dr.Y Kertih) to utilize several organic
xenobiotics such as phenol, acrylamide, nicotinamide, acetamide, iodoacetamide, propionamide,
acetamide, sodium dodecyl sulfate (SDS) and diesel as electron donor sources for aiding
reduction or as carbon sources for growth was also examined. Finding showed that none was
capable of aiding molybdenum reduction, however the bacterium was capable of growing on both
diesel and phenol as carbon sources. GC analysis was used to confirmed diesel degradation.
The application of chemical dispersants in marine oil spill remediation is comprehensively reported across the globe. But, the augmented toxicity and poor biodegradability of reported chemical dispersants have created necessity for their replacement with the bio-based green dispersants. Therefore, in the present study, we have synthesized five ionic liquids (ILs) namely 1-butyl-3-methylimidazolium lauroylsarcosinate, 1,1'-(1,4-butanediyl)bis(1-H-pyrrolidinium) dodecylbenzenesulfonate, tetrabutylammonium citrate, tetrabutylammonium polyphosphate and tetrabutylammonium ethoxylate oleyl ether glycolate, and formulated a water based ILs dispersant combining the synthesized ILs at specified compositions. The effectiveness of formulated ILs dispersant was found between 70.75% and 94.71% for the dispersion of various crude oils ranging from light to heavy. Further, the acute toxicity tests against zebra fish and grouper fish have revealed the practically non-toxic behaviour of formulated ILs dispersant with LC50 value greater than 100 ppm after 96 h. In addition, the formulated ILs dispersant has provided excellent biodegradability throughout the test period. Overall, the formulated new ILs dispersant is deemed to facilitate environmentally benign oil spill remediation and could effectively substitute the use of hazardous chemical dispersants in immediate future.
Hydrazine is an intermediate product of the anaerobic ammonium oxidation (Anammox) process where both ammonium and nitrite in wastewater are converted to nitrogen gas by bacteria. In this study the effect of external hydrazine addition (5, 10, 15, and 20 mg/L) on the start-up period of the Anammox process was studied using sequencing batch reactors (SBRs). The SBR with an addition of 10 mg/L hydrazine took only 7 weeks to stabilize and achieve the maximum removal of ammonium and nitrite, whereas the SBR without the addition of hydrazine took 12 weeks. The amount of Heme C extracted from the biomass indicated that externally added hydrazine accelerated the growth of Anammox bacteria and reduced the release of nitrous oxide gas from the reactors.
A glutathione S-transferase (GST) with a potential dehalogenation function against various organochlorine substrates was identified from a polychlorobiphenyl (PCB)-degrading organism, Acidovorax sp. KKS102. A homolog of the gene BphK (biphenyl upper pathway K), named BphK-KKS, was cloned, purified and biochemically characterized. Bioinformatic analysis indicated several conserved amino acids that participated in the catalytic activity of the enzyme, and site-directed mutagenesis of these conserved amino acids revealed their importance in the enzyme's catalytic activity. The wild-type and mutant (C10F, K107T and A180P) recombinant proteins displayed wider substrate specificity. The wild-type recombinant GST reacted towards 1-chloro-2,4-dinitrobenzene (CDNB), ethacrynic acid, hydrogen peroxide and cumene hydroperoxide. The mutated recombinant proteins, however, showed significant variation in specific activities towards the substrates. A combination of a molecular docking study and a chloride ion detection assay showed potential interaction with and a dechlorination function against 2-, 3- and 4-chlorobenzoates (metabolites generated during PCB biodegradation) in addition to some organochlorine pesticides (dichlorodiphenyltrichloroethane, endosulfan and permethrin). It was demonstrated that the behavior of the dechlorinating activities varied among the wild-type and mutant recombinant proteins. Kinetic studies (using CDNB and glutathione) showed that the kinetic parameters Km, Vmax, Kcat and Km/Kcat were all affected by the mutations. While C10F and A180P mutants displayed an increase in GST activity and the dechlorination function of the enzyme, the K107T mutant displayed variable results, suggesting a functional role of Lys107 in determining substrate specificity of the enzyme. These results demonstrated that the enzyme should be valuable in the bioremediation of metabolites generated during PCB biodegradation.
The industrial contamination of marine sediments with mercury, silver, and zinc in Penang, Malaysia was studied with bio-remediation coupled with power generation using membrane less open (aerated) and closed (non-aerated) sediment microbial fuel cells (SMFCs). The prototype for this SMFC is very similar to a natural aquatic environment because it is not stimulated externally and an oxygen sparger is inserted in the cathode chamber to create the aerobic environment in the open SMFC and no oxygen supplied in the closed SMFC. The open and closed SMFCs were showed the maximum voltage generation 300.5 mV (77.75 mW/m2) and 202.7 mV (45.04 (mW/m2), respectively. The cyclic voltammetry showed the oxidation peak in open SMFCs at +1.9 μA and reduction peak at -0.3 μA but in closed SMFCs oxidation and reduction peaks were noted at +1.5 μA and -1.0 μA, respectively. The overall impedance (anode, cathode and solution) of closed SMFCs was higher than open SMFCs. The charge transfer impedance showed that the rates of substrate oxidation and reduction were very low in the closed SMFCs than open SMFCs. The Nyquist arc indicated that O2 act as electron acceptor in the open SMFCs and CO2 in the closed SMFCs. The highest remediation efficiency of toxic metals [Hg (II) ions, Zn (II) ions, and Ag (I) ions] in the open SMFCs were 95.03%, 86.69%, and 83.65% in closed SMFCs were 69.53%, 66.57%, and 65.33%, respectively, observed during 60-80 days. The scanning electron microscope and 16S rRNA analysis showed diverse exoelectrogenic community in the open SMFCs and closed SMFCs. The results demonstrated that open SMFCs could be employed for the power generation and bioremediation of pollutants.
Endophytic fungi provide protection to their host plant and the fungi often produce antimicrobial compounds to aid the host
fighting off pathogens. These bioactive compounds were secondary metabolites which were often produced as waste- or
by-products. In the present study, endophytic fungi isolated from mangrove plants and soils were characterized and their
antimicrobial production and bioremediation potential of heavy metals copper (Cu) and zinc (Zn) were assessed. Twelve
(12) isolated and identified endophytic fungi belonged to seven species; Penicillium, Curvularia, Diaporthe, Aspergillus,
Guignardia, Neusartorya and Eupenicillium. Antimicrobial activities of these 12 fungal endophytes were tested against
Gram negative bacteria; Bacillus subtilis, Staphylococcus aureus, Gram positive bacteria; Escherichia coli and fungi;
Candida albicans and Aspergillus niger among others. Two isolates (related to Guignardia sp. and Neusartoya sp.) showed
strong antimicrobial (and antifungal) activity whereas the rest showed no activity. Compounds were isolated from both
isolates and screened using HPLC. Both isolates displayed chemically very interesting chromatograms as they possessed a
high diversity of basic chemical structures and peaks over a wide range of polarities, with structures similar to Trimeric
catechin and Helenalin among others. For bioremediation assessment, the results showed maximum biosorption capacity
for two isolates related to Curvularia sp. and Neusartorya sp., with the former removing 25 mg Cu/g biomass and the
latter removing 24 mg Zn/g biomass. Our results indicated the potential of mangrove endophytic fungi in producing
bioactive compounds and also highlighted their potential for the treatment of heavy metal-contaminated wastewater.
Microalgal-bacterial granular sludge (MBGS) process has become a focal point in treating municipal wastewater. However, it remains elusive whether the emerging process can be applied for the treatment of aquaculture wastewater, which contains considerable concentrations of nitrate and nitrite. This study evaluated the feasibility of MBGS process for aquaculture wastewater treatment. Result showed that the MBGS process was competent to remove respective 64.8%, 84.9%, 70.8%, 50.0% and 84.2% of chemical oxygen demand, ammonia-nitrogen, nitrate-nitrogen, nitrite-nitrogen and phosphate-phosphorus under non-aerated conditions within 8 h. The dominant microalgae and bacteria were identified to be Coelastrella and Rhodobacteraceae, respectively. Further metagenomics analysis implied that microbial assimilation was the main contributor in organics, nitrogen and phosphorus removal. Specifically, considerable nitrate and nitrite removals were also obtained with the synergy between microalgae and bacteria. Consequently, this work demonstrated that the MBGS process showed a prospect of becoming an environmentally friendly and efficient alternative in aquaculture wastewater treatment.
In recent years, the utilisation of endophytes has emerged as a promising biological treatment technology for the degradation of plastic wastes such as biodegradation of synthetic plastics. This study, therefore, aimed to explore and extensively screen endophytic fungi (from selected plants) for efficient in vitro polyvinyl alcohol (PVA) biodegradation. In total, 76 endophytic fungi were isolated and cultivated on a PVA screening agar medium. Among these fungi, 10 isolates showed potential and were subsequently identified based on phenotypical characteristics, ITS ribosomal gene sequences, and phylogenetic analyses. Four strains exhibited a maximum level of PVA-degradation in the liquid medium when cultivated for 10 days at 28 °C and 150 rpm. These strains showed varied PVA removal rates of 81% (Penicillium brevicompactum OVR-5), 67% (Talaromyces verruculosus PRL-2), 52% (P. polonicum BJL-9), and 41% (Aspergillus tubingensis BJR-6) respectively. The most promising PVA biodegradation isolate 'OVR-5', with an optimal pH at 7.0 and optimal temperature at 30 °C, produced lipase, manganese peroxidase, and laccase enzymes. Based on analyses of its metabolic intermediates, as identified with GC-MS, we proposed the potential PVA degradation pathway of OVR-5. Biodegradation results were confirmed through scanning electron microscopy and Fourier transform infrared spectroscopy. This study provides the first report on an endophytic P. brevicompactum strain (associated with Orychophragmus violaceus) that has a great ability for PVA degradation providing more insight on potential fungus-based applications in plastic waste degradation.
Liquid detergent has an increasing demand in North America, Western Europe, and Southeast Asia countries owing to its convenience to use and efficiency to clean. Alpha methyl ester sulfonates (α-MES), an anionic surfactant derived from palm oil based methyl ester, was reported to have lower manufacturing cost, good detergency with less dosage, excellent biodegradability, higher tolerance to hard water, and lower eco-toxicity as compared to linear alkylbenzene sulfonates (LABS). LABS was known as the workhorse of the detergent industry in the 20th century. Although palm-based α-MES was successfully used as the sole surfactant in powder detergent, there are still some unsettled technical issues related to phase stability and viscosity when using this anionic surfactant in heavy-duty laundry liquid detergent formulations. This paper will review not only the market overview of detergents, the application and performance of green surfactants in laundry detergents but also will highlight the technical issues related to the application of palm-based α-MES in laundry liquid detergent and some of the possible methods to overcome the formulation adversities.
The continuous accumulation of microplastics in the environment poses ecological threats and has been an increasing problem worldwide. In this study, eight bacterial strains were isolated from mangrove sediment in Peninsular Malaysia to mitigate the environmental impact of microplastics and develop a clean-up option. The bacterial isolates were screened for their potential to degrade UV-treated microplastics from polyethylene (PE), polyethylene terephthalate (PET), polypropylene (PP), and polystyrene (PS). Only two isolates, namely, Bacillus cereus and Bacillus gottheilii, grew on a synthetic medium containing different microplastic polymers as the sole carbon source. A shake flask experiment was carried out to further evaluate the biodegradability potential of the isolates. Degradation was monitored by recording the weight loss of microplastics and the growth pattern of the isolates in the mineral medium. The biodegradation extent was validated by assessment of the morphological and structural changes through scanning electron microscopy and Fourier transform infrared spectroscopy analyses. The calculated weight loss percentages of the microplastic particles by B. cereus after 40 days were 1.6%, 6.6%, and 7.4% for PE, PET, and PS, respectively. B. gottheilii recorded weight loss percentages of 6.2%, 3.0%, 3.6%, and 5.8% for PE, PET, PP, and PS, respectively. The designated isolates degraded the microplastic material and exhibited potential for remediation of microplastic-contaminated environment. Biodegradation tests must be conducted to characterize the varied responses of microbes toward pollutants, such as microplastics. Hence, a novel approach for biodegradation of microplastics must be developed to help mitigate the environmental impact of plastics and microplastic polymers.
Interest in the biodegradation of microplastics is due to their ubiquitous distribution, availability, high persistence in the environment and deleterious impact on marine biota. The present study evaluates the growth response and mechanism of polypropylene (PP) degradation by Bacillus sp. strain 27 and Rhodococcus sp. strain 36 isolated from mangrove sediments upon exposure to PP microplastics. Both bacteria strains were able to utilise PP microplastic for growth as confirmed by the reduction of the polymer mass. The weight loss was 6.4% by Rhodococcus sp. strain 36 and 4.0% by Bacillus sp. strain 27 after 40days of incubation. PP biodegradation was further confirmed using Fourier-transform infrared spectroscopy and scanning electron microscopy analyses, which revealed structural and morphological changes in the PP microplastics with microbial treatment. These analyses showed that the isolates can colonise, modify and utilise PP microplastics as carbon source.
While the oil palm industry has been recognized for its contribution towards economic growth and rapid development, it has also contributed to environmental pollution due to the production of huge quantities of by-products from the oil extraction process. A phytoremediation technique (floating Vetiver system) was used to treat Palm Oil Mill Secondary Effluent (POMSE). A batch study using 40 L treatment tanks was carried out under different conditions and Response Surface Methodology (RSM) was applied to optimize the treatment process. A three factor central composite design (CCD) was used to predict the experimental variables (POMSE concentration, Vetiver plant density and time). An extraordinary decrease in organic matter as measured by BOD and COD (96% and 94% respectively) was recorded during the experimental duration of 4 weeks using a density of 30 Vetiver plants. The best and lowest final BOD of 2 mg/L was obtained when using 15 Vetiver plants after 13 days for low concentration POMSE (initial BOD = 50 mg/L). The next best result of BOD at 32 mg/L was obtained when using 30 Vetiver plants after 24 days for medium concentration POMSE (initial BOD = 175 mg/L). These results confirmed the validity of the model, and the experimental value was determined to be quite close to the predicted value, implying that the empirical model derived from RSM experimental design can be used to adequately describe the relationship between the independent variables and response. The study showed that the Vetiver system is an effective method of treating POMSE.
Benzo[a]pyrene is a high-molecular-weight polycyclic aromatic hydrocarbon highly recalcitrant in nature and thus harms the ecosystem and/or human health. Therefore, its removal from the marine environment is crucial. This research focuses on benzo[a]pyrene degradation by using enriched bacterial isolates in consortium under saline conditions. Bacterial isolates capable of using benzo[a]pyrene as sole source of carbon and energy were isolated from enriched mangrove sediment. These isolates were identified as Ochrobactrum anthropi, Stenotrophomonas acidaminiphila, and Aeromonas salmonicida ss salmonicida. Isolated O. anthropi and S. acidaminiphila degraded 26% and 20%, respectively, of an initial benzo[a]pyrene concentration of 20 mg/L after 8 days of incubation in seawater (28 ppm of NaCl). Meanwhile, the bacterial consortium decomposed 41% of an initial 50 mg/L benzo[a]pyrene concentration after 8 days of incubation in seawater (28 ppm of NaCl). The degradation efficiency of benzo[a]pyrene increased to 54%, when phenanthrene was supplemented as a co-metabolic substrate. The order of biodegradation rate by temperature was 30°C > 25°C > 35°C. Our results suggest that co-metabolism by the consortium could be a promising biodegradation strategy for benzo[a]pyrene in seawater.