Numerous technologies and approaches have been used in the past few decades to remove hexavalent chromium (Cr[VI]) in wastewater and the environment. However, these conventional technologies are not economical and efficient in removing Cr(VI) at a very low concentration (1-100 ppm). As an alternative, the utilization of bioremediation techniques which uses the potential of microorganisms could represent an effective technique for the detoxification of Cr(VI). In this study, we reported a newly isolated bacterium identified as Acinetobacter radioresistens sp. NS-MIE from Malaysian agricultural soil. The chromate reduction potential of strain NS-MIE was optimized using RSM and ANN techniques. The optimum condition predicted by RSM for the bacterium to reduce hexavalent chromium occurred at pH 6, 10 g/L ppm of nutrient broth (NB) concentration and 100 ppm of chromate concentration while the optimum condition predicted by ANN is at pH 6 and 10 g/L of NB concentration and of 60 ppm of chromate concentration with chromate reduction (%) of 75.13 % and 96.27 %, respectively. The analysis by the ANN model shows better prediction data with a higher R2 value of 0.9991 and smaller average absolute deviation (AAD) and root mean square error (RMSE) of 0.33 % and 0.302 %, respectively. Validation analysis showed the predicted values by RSM and ANN were close to the validation values, whereas the ANN showed the lowest deviation, 2.57%, compared to the RSM. This finding suggests that the ANN showed a better prediction and fitting ability compared to the RSM for the nonlinear regression analysis. Based on this study, A. radioresistens sp. NS-MIE exhibits strong potential characteristics as a candidate for the bioremediation of hexavalent chromium in the environment.
This study investigates the ultrasound-assisted extraction of flavonoids from Malaysian cocoa shell extracts, and optimization using response surface methodology. There are three variables involved in this study, namely: ethanol concentration (70⁻90 v/v %), temperature (45⁻65 °C), and ultrasound irradiation time (30⁻60 min). All of the data were collected and analyzed for variance (ANOVA). The coefficient of determination (R²) and the model was significant in interaction between all variables (98% and p < 0.0001, respectively). In addition, the lack of fit test for the model was not of significance, with p > 0.0684. The ethanol concentration, temperature, and ultrasound irradiation time that yielded the maximum value of the total flavonoid content (TFC; 7.47 mg RE/g dried weight (DW)) was 80%, 55 °C, and 45 min, respectively. The optimum value from the validation of the experimental TFC was 7.23 ± 0.15 mg of rutin, equivalent per gram of extract with ethanol concentration, temperature, and ultrasound irradiation time values of 74.20%, 49.99 °C, and 42.82 min, respectively. While the modelled equation fits the data, the T-test is not significant, suggesting that the experimental values agree with those predicted by the response surface methodology models.
Potentially toxic metals pollution in the Straits of Malacca warrants the development of rapid, simple and sensitive assays. Enzyme-based assays are excellent preliminary screening tools with near real-time potential. The heavy-metal assay based on the protease ficin was optimized for mercury detection using response surface methodology. The inhibitive assay is based on ficin action on the substrate casein and residual casein is determined using the Coomassie dye-binding assay. Toxic metals strongly inhibit this hydrolysis. A central composite design (CCD) was utilized to optimize the detection of toxic metals. The results show a marked improvement for the concentration causing 50% inhibition (IC50) for mercury, silver and copper. Compared to one-factor-at-a-time (OFAT) optimization, RSM gave an improvement of IC50 (mg/L) from 0.060 (95% CI, 0.030-0.080) to 0.017 (95% CI, 0.016-0.019), from 0.098 (95% CI, 0.077-0.127) to 0.028 (95% CI, 0.022-0.037) and from 0.040 (95% CI, 0.035-0.045) to 0.023 (95% CI, 0.020-0.027), for mercury, silver and copper, respectively. A near-real time monitoring of mercury concentration in the Straits of Malacca at one location in Port Klang was carried out over a 4 h interval for a total of 24 h and validated by instrumental analysis, with the result revealing an absence of mercury pollution in the sampling site.
Predicting the crucial effect of single metal pollutants against the aquatic ecosystem has been highly debatable for decades. However, dealing with complex metal mixtures management in toxicological studies creates a challenge, as heavy metals may evoke greater toxicity on interactions with other constituents rather than individually low acting concentrations. Moreover, the toxicity mechanisms are different between short term and long term exposure of the metal toxicant. In this study, acute and chronic toxicity based on luminescence inhibition assay using newly isolated Photobacterium sp.NAA-MIE as the indicator are presented. Photobacterium sp.NAA-MIE was exposed to the mixture at a predetermined ratio of 1:1. TU (Toxicity Unit) and MTI (Mixture Toxic Index) approach presented the mixture toxicity of Hg2+ + Ag+, Hg2+ + Cu2+, Ag+ + Cu2+, Hg2+ + Ag+ + Cu2+, and Cd2+ + Cu2+ showed antagonistic effect over acute and chronic test. Binary mixture of Cu2+ + Zn2+ was observed to show additive effect at acute test and antagonistic effect at chronic test while mixture of Ni2+ + Zn2+ showing antagonistic effect during acute test and synergistic effect during chronic test. Thus, the strain is suitable and their use as bioassay to predict the risk assessment of heavy metal under acute toxicity without abandoning the advantage of chronic toxicity extrapolation.
Assessment of eco-toxicant using bioluminescent bacterial assay is a widely used and globally accepted method. In this work, a new luminescent bacterium was isolated from squid (Loligo duvauceli) and identified as Photobacterium leiognathi strain AK-MIE using 16S rRNA, phylogeny analysis. The predicted optimum conditions by RSM were 2.76% (w/v) NaCl, 2.28% (w/v) peptone, 0.34% (w/v) yeast extract, and pH 6.83 with 541,211.80 RLU of luminescent production whereas the predicted optimum conditions by ANN were 2.21% (w/v) NaCl, 2.27% (w/v) peptone, 0.39% (w/v) yeast extract, and pH 6.94 which produced 541,986.20 RLU. The validation analysis of both RSM and ANN show 0.60% and 0.69% deviation from the predicted results indicating that both models provided good quality predictions with ANN showing a superior data fitting capability for non-linear regression analysis. Toxicity tests show strain AK-MIE was sensitive to mercury (concentration causing 50% inhibition or IC50 of 0.00978 mgL-1), followed by cadmium (IC50 of 0.5288 mgL-1), copper IC50 of (0.8117 mgL-1), silver (IC50 of 1.109 mgL-1), and lead (IC50 of 10.71 mgL-1) which are more sensitive than previously isolated luminescent bacteria, suggesting that strain AK-MIE has the potential to be used in toxicity assessment of heavy metals in the environment. Based on the field trial results, several sediment samples from industrial areas in Bangi, Selangor managed to inhibit the bioluminescence of strain AK-MIE. Validation method carried out using ICP-MS proved the presence of several toxic heavy metal elements.
The present study was conducted to optimize extraction process for defatted pitaya seed extract (DPSE) adopting response surface methodology (RSM). A five-level central composite design was used to optimize total phenolic content (TPC), total flavonoid content (TFC), ferric reducing antioxidant power (FRAP), and 2,2'-azino-bis (3-ethylbenzothizoline-6-sulfonic acid (ABTS) activities. The independent variables included extraction time (30-60 min), extraction temperature (40-80 °C) and ethanol concentration (60%-80%). Results showed that the quadratic polynomial equations for all models were significant at (p < 0.05), with non-significant lack of fit at p > 0.05 and R2 of more than 0.90. The optimized extraction parameters were established as follows: extraction time of 45 min, extraction temperature of 70 °C and ethanol concentration of 80%. Under these conditions, the recovery of TPC, TFC, and antioxidant activity based on FRAP and ABTS were 128.58 ± 1.61 mg gallic acid equivalent (GAE)/g sample, 9.805 ± 0.69 mg quercetin equivalent (QE)/g sample, 1.23 ± 0.03 mM Fe2+/g sample, and 91.62% ± 0.15, respectively. Ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) analysis identified seven chemical compounds with flavonoids constituting major composition of the DPSE.
In this study, the optimal conditions for the extraction of antioxidants from the Buah Mahkota Dewa fruit (Phaleria macrocarpa) was determined by using Response Surface Methodology (RSM). The optimisation was applied using a Central Composite Design (CCD) to investigate the effect of three independent variables, namely extraction temperature (°C), extraction time (minutes) and extraction solvent to-feed ratio (% v/v) on four responses: free radical scavenging activity (DPPH), ferric ion reducing power assay (FRAP), total phenolic content (TPC) and total flavonoid content (TFC). The optimal conditions for the antioxidants extraction were found to be 64 °C extraction temperature, 66 min extraction time and 75% v/v solvent to-feed ratio giving the highest percentage yields of DPPH, FRAP, TPC and TFC of 86.85%, 7.47%, 292.86 mg/g and 3.22 mg/g, respectively. Moreover, the data were subjected to Response Surface Methodology (RSM) and the results showed that the polynomial equations for all models were significant, did not show lack of fit, and presented adjusted determination coefficients (R²) above 99%, proving that the yield of phenolic, flavonoid and antioxidants activities obtained experimentally were close to the predicted values and the suitability of the model employed in RSM to optimise the extraction conditions. Hence, in this study, the fruit from P. macrocarpa could be considered to have strong antioxidant ability and can be used in various cosmeceutical or medicinal applications.
Currently, consumers' demand for sunscreens derived from natural sources that provide photoprotection from ultraviolet (UV) radiation is pushing the cosmetic industry to develop breakthrough formulations of sun protection products by incorporating plant antioxidants as their active ingredients. In this context, the present study was initiated to evaluate the antioxidant and photoprotective properties of the underutilized Hylocereus polyrhizus peel extract (HPPE) using in vitro spectrophotometric techniques. The phytochemical screenings of HPPE conducted via high-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) revealed the presence of phenolic acids and flavonoids as the major secondary metabolites in HPPE. The antioxidant potentials evaluated based on 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical and total antioxidant capacity assays were in the range of 22.16 ± 0.24%-84.67 ± 0.03% with 50% inhibitory concentration (IC50) of 36.39 ± 0.04 μg/mL and 23.76 ± 0.14%-31.87 ± 0.26% (IC50 = 21.93 ± 0.07 μg/mL), respectively. For the photoprotective evaluation, the results showed that HPPE had significantly high absorbance values (3.1-3.6) at 290-320 nm with an exceptional sun protection factor (SPF) value of 35.02 ± 0.39 at 1.00 mg/mL. HPPE also possessed a broad-spectrum shielding power against both UVA and UVB radiations. Hence, in terms of practical implications, our findings would offer an exciting avenue to develop a photoprotective formulation incorporating the ethanolic extract of Hylocereus polyrhizus peels as a synergistic active ingredient for its excellent UV absorption properties and the strong antioxidant activities.
Most components of petroleum oily sludge (POS) are toxic, mutagenic and cancer-causing. Often bioremediation using microorganisms is hindered by the toxicity of POS. Under this circumstance, phytoremediation is the main option as it can overcome the toxicity of POS. Cajanus cajan a legume plant, was evaluated as a phyto-remediating agent for petroleum oily sludge-spiked soil. Culture dependent and independent methods were used to determine the rhizosphere microorganisms' composition. Degradation rates were estimated gravimetrically. The population of total heterotrophic bacteria (THRB) was significantly higher in the uncontaminated soil compared to the contaminated rhizosphere soil with C. cajan, but the population of hydrocarbon-utilizing bacteria (HUB) was higher in the contaminated rhizosphere soil. The results show that for 1 to 3% oily sludge concentrations, an increase in microbial counts for all treatments from day 0 to 90 d was observed with the contaminated rhizosphere CR showing the highest significant increase (p
The application of microorganisms in azo dye remediation has gained significant attention, leading to various published studies reporting different methods for obtaining the best dye decolouriser. This paper investigates and compares the role of methods and media used in obtaining a bacterial consortium capable of decolourising azo dye as the sole carbon source, which is extremely rare to find. It was demonstrated that a prolonged acclimation under low substrate availability successfully isolated a novel consortium capable of utilising Reactive Red 120 dye as a sole carbon source in aerobic conditions. This consortium, known as JR3, consists of Pseudomonas aeruginosa strain MM01, Enterobacter sp. strain MM05 and Serratia marcescens strain MM06. Decolourised metabolites of consortium JR3 showed an improvement in mung bean's seed germination and shoot and root length. One-factor-at-time optimisation characterisation showed maximal of 82.9% decolourisation at 0.7 g/L ammonium sulphate, pH 8, 35 °C, and RR120 concentrations of 200 ppm. Decolourisation modelling utilising response surface methodology (RSM) successfully improved decolourisation even more. RSM resulted in maximal decolourisation of 92.79% using 0.645 g/L ammonium sulphate, pH 8.29, 34.5 °C and 200 ppm RR120.
A very sensitive and selective colorimetric biosensor for the measurement of mercury ion (Hg2+) in environmental samples has been developed using functionalized gold nanoparticles with bromelain enzyme (brn-AuNPs). This work has shown that Hg2+ measurement based on spectrophotometer and digital image analysis is a very innovative and successful method for providing an effective preliminary system and has promise for the future of water quality biomonitoring. Response Surface Methodology (RSM), a Box-Behnken design-based technique, was used to identify the optimum levels of functionalization of bromelain to AuNPs. The created model's validity was confirmed, and statistical analysis revealed that the ideal functionalize conditions were 1 mM of AuNPs, functionalize with 0.59 mM bromelain concentration on 14 ℃ temperature and 72 h incubation time. The lowest colorimetric detection concentration (LOD) of brn-AuNPs of Hg2+ was 0.0092 ppm and 0.011 ppm for spectrophotometer and digital image analysis. As shown, digital image analysis had advantages based on the LOD result comparable to UV-VIS spectrophotometer. The practical application of the brn-AuNPs sensing was proven with mercury determination in water samples. The present study developed a robust sensor, which successfully implemented in a compact portable sensor kit, turning this sensor into a very potent tool for the development water quality biomonitoring system of Hg2+ application.
The reason for such enormous efforts in palm oil mill effluent research would be what has been singled out as one of the major sources of pollution in Malaysia, and perhaps the most costly and complex waste to manage. Palm oil mill final discharge, which is the treated effluent, will usually be discharged to nearby land or river since it has been the least costly way to dispose of. Irrefutably, the quality level of the treated effluent does not always satisfy the surface water quality in conformity to physicochemical characteristics. To work on improving the treated effluent quality, a vertical surface-flow constructed wetland system was designed with Pennisetum purpureum (Napier grass) planted on the wetland floor. The system effectively reduced the level of chemical oxygen demand by 62.2 ± 14.3%, total suspended solid by 88.1 ± 13.3%, ammonia by 62.3 ± 24.8%, colour by 66.6 ± 13.19%, and tannin and lignin by 57.5 ± 22.3%. Heat map depicted bacterial diversity and relative abundance in life stages from the wetland soil, whereby bacterial community associated with the pollutant removal was found to be from the families Anaerolineaceae and Nitrosomonadaceae, and phyla Cyanobacteria and Acidobacteria.
Constructed wetlands (CWs) are affordable and reliable green technologies for the treatment of various types of wastewater. Compared to conventional treatment systems, CWs offer an environmentally friendly approach, are low cost, have fewer operational and maintenance requirements, and have a high potential for being applied in developing countries, particularly in small rural communities. However, the sustainable management and successful application of these systems remain a challenge. Therefore, after briefly providing basic information on wetlands and summarizing the classification and use of current CWs, this study aims to provide and inspire sustainable solutions for the performance and application of CWs by giving a comprehensive review of CWs' application and the recent development of their sustainable design, operation, and optimization for wastewater treatment. To accomplish this objective, thee design and management parameters of CWs, including macrophyte species, media types, water level, hydraulic retention time (HRT), and hydraulic loading rate (HLR), are discussed. Besides these, future research on improving the stability and sustainability of CWs are highlighted. This article provides a tool for researchers and decision-makers for using CWs to treat wastewater in a particular area. This paper presents an aid for informed analysis, decision-making, and communication. The review indicates that major advances in the design, operation, and optimization of CWs have greatly increased contaminant removal efficiencies, and the sustainable application of this treatment system has also been improved.