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The first phase of toxicity identification evaluation (TIE) method comprised of physic-chemical fractionation steps of pH adjustment, pH adjustment followed by filtration, aeration, extraction with solid phase C18 column (SPE), oxidant reduction with sodium thiosulphate and EDTA chelation was conducted to characterize the toxicants of a Malaysian landfill leachate. The battery of organisms test chosen were freshwater fish (Rasbora sumatrana), freshwater prawn (Macrobrachium lanchesteri) and tomato seed (Lycoperson esculentum). Toxicity reductions at each step were comparable for all test organisms. The major toxicants present in the leachate were found to be mostly basic in nature and precipitable under acidic conditions as well as containing non-polar organic compounds. A small reduction in toxicity was observed when leachate was treated with sodium thiosulphate in oxidant reduction test indicating the presence of oxidizers. The EDTA chelating step did not significantly reduce toxicity in the test organisms suggesting insignificant level of (toxic) metals.

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A fluorescence-based fiber optic toxicity biosensor based on genetically modified Escherichia coli (E. coli) with green fluorescent protein (GFP) was developed for the evaluation of the toxicity of several hazardous heavy metal ions. The toxic metals include Cu(II), Cd(II), Pb(II), Zn(II), Cr(VI), Co(II), Ni(II), Ag(I) and Fe(III). The optimum fluorescence excitation and emission wavelengths of the optical biosensor were 400 ± 2 nm and 485 ± 2 nm, respectively. Based on the toxicity observed under optimal conditions, the detection limits of Cu(II), Cd(II), Pb(II), Zn(II), Cr(VI), Co(II), Ni(II), Ag(I) and Fe(III) that can be detected using the toxicity biosensor were at 0.04, 0.32, 0.46, 2.80, 100, 250, 400, 720 and 2600 µg/L, respectively. The repeatability and reproducibility of the proposed biosensor were 3.5%-4.8% RSD (relative standard deviation) and 3.6%-5.1% RSD (n = 8), respectively. The biosensor response was stable for at least five weeks, and demonstrated higher sensitivity towards metal toxicity evaluation when compared to a conventional Microtox assay.

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In this article a luminescence fiber optic biosensor for the microdetection of heavy metal toxicity in waters based on the marine bacterium Aliivibrio fischeri (A. fischeri) encapsulated in alginate microspheres is described. Cu(II), Cd(II), Pb(II), Zn(II), Cr(VI), Co(II), Ni(II), Ag(I) and Fe(II) were selected as sample toxic heavy metal ions for evaluation of the performance of this toxicity microbiosensor. The loss of bioluminescence response from immobilized A. fischeri bacterial cells corresponds to changes in the toxicity levels. The inhibition of the luminescent biosensor response collected at excitation and emission wavelengths of 287 ± 2 nm and 487 ± 2 nm, respectively, was found to be reproducible and repeatable within the relative standard deviation (RSD) range of 2.4-5.7% (n = 8). The toxicity biosensor based on alginate micropsheres exhibited a lower limit of detection (LOD) for Cu(II) (6.40 µg/L), Cd(II) (1.56 µg/L), Pb(II) (47 µg/L), Ag(I) (18 µg/L) than Zn(II) (320 µg/L), Cr(VI) (1,000 µg/L), Co(II) (1700 µg/L), Ni(II) (2800 µg/L), and Fe(III) (3100 µg/L). Such LOD values are lower when compared with other previous reported whole cell toxicity biosensors using agar gel, agarose gel and cellulose membrane biomatrices used for the immobilization of bacterial cells. The A. fischeri bacteria microencapsulated in alginate biopolymer could maintain their metabolic activity for a prolonged period of up to six weeks without any noticeable changes in the bioluminescence response. The bioluminescent biosensor could also be used for the determination of antagonistic toxicity levels for toxicant mixtures. A comparison of the results obtained by atomic absorption spectroscopy (AAS) and using the proposed luminescent A. fischeri-based biosensor suggests that the optical toxicity biosensor can be used for quantitative microdetermination of heavy metal toxicity in environmental water samples.

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Tourism-related activities such as the heavy use of boats for transportation are a significant source of petroleum hydrocarbons that may harm the ecosystem of Langkawi Island. The contamination and toxicity levels of polycyclic aromatic hydrocarbon (PAH) in the sediments of Langkawi were evaluated using sediment quality guidelines (SQGs) and toxic equivalent factors. Ten samples were collected from jetties and fish farms around the island in December 2010. A gas chromatography/flame ionization detector (GC/FID) was used to analyse the 18 PAHs. The concentration of total PAHs was found to range from 869 ± 00 to 1637 ± 20 ng g⁻¹ with a mean concentration of 1167.00 ± 24 ng g⁻¹, lower than the SQG effects range-low (3442 ng g⁻¹). The results indicated that PAHs may not cause acute biological damage. Diagnostic ratios and principal component analysis suggested that the PAHs were likely to originate from pyrogenic and petrogenic sources. The toxic equivalent concentrations of the PAHs ranged from 76.3 to 177 ng TEQ/g d.w., which is lower compared to similar studies. The results of mean effects range-median quotient of the PAHs were lower than 0.1, which indicate an 11% probability of toxicity effect. Hence, the sampling sites were determined to be the low-priority sites.

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Whole cell biosensors always face the challenge of low stability of biological components and short storage life. This paper reports the effects of poly(2-hydroxyethyl methacrylate) (pHEMA) immobilization on a whole cell fluorescence biosensor for the detection of heavy metals (Cu, Pb, Cd), and pesticides (dichlorophenoxyacetic acid (2,4-D), and chlorpyrifos). The biosensor was produced by entrapping the cyanobacterium Anabaena torulosa on a cellulose membrane, followed by applying a layer of pHEMA, and attaching it to a well. The well was then fixed to an optical probe which was connected to a fluorescence spectrophotometer and an electronic reader. The optimization of the biosensor using several factors such as amount of HEMA and drying temperature were undertaken. The detection limits of biosensor without pHEMA for Cu, Cd, Pb, 2,4-D and chlorpyrifos were 1.195, 0.027, 0.0100, 0.025 and 0.025 µg/L respectively. The presence of pHEMA increased the limits of detection to 1.410, 0.250, 0.500, 0.235 and 0.117 µg/L respectively. pHEMA is known to enhance the reproducibility of the biosensor with average relative standard deviation (RSD) of ±1.76% for all the pollutants tested, 48% better than the biosensor without pHEMA (RSD = ±3.73%). In storability test with Cu 5 µg/L, the biosensor with pHEMA performed 11.5% better than the test without pHEMA on day-10 and 5.2% better on day-25. pHEMA is therefore a good candidate to be used in whole cell biosensors as it increases reproducibility and enhances biosensor storability.

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The palm oil industry has played an important role in the economic development of Malaysia and has enhanced the economic welfare of its people. To determine the environmental impact of the oil palm seedling at the nursery stage, information on inputs and outputs need to be assessed. The oil palm nursery is the first link in the palm oil supply chain. A gate-to-gate study was carried out whereby the system boundary was set to only include the process of the oil palm seedling. The starting point was a germinated seed in a small polyethylene bag (6 in × 9 in) in which it remained until the seedling was approximately 3 to 4 months old. The seedling was then transferred into a larger polyethylene bag (12 in × 15 in), where it remained until it was 10-12 months old, when it was planted in the field (plantation). The functional unit for this life cycle inventory (LCI) is based on the production of one seedling. Generally, within the system boundary, the production of an oil palm seedling has only two major environmental impact points, the polybags used to grow the seedling and the fungicide (dithiocarbamate) used to control pathogenic fungi, as both the polybags and the dithiocarbamate are derived from fossil fuel.

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The growing interest in the environmental occurrence of veterinary and human pharmaceuticals is essentially due to their possible health implications to humans and ecosystem. This study assesses the occurrence of human pharmaceuticals in a Malaysian tropical aquatic environment taking a chemometric approach using cluster analysis, discriminant analysis and principal component analysis. Water samples were collected from seven sampling stations along the heavily populated Langat River basin on the west coast of peninsular Malaysia and its main tributaries. Water samples were extracted using solid-phase extraction and analyzed using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for 18 pharmaceuticals and one metabolite, which cover a range of six therapeutic classes widely consumed in Malaysia. Cluster analysis was applied to group both pharmaceutical pollutants and sampling stations. Cluster analysis successfully clustered sampling stations and pollutants into three major clusters. Discriminant analysis was applied to identify those pollutants which had a significant impact in the definition of clusters. Finally, principal component analysis using a three-component model determined the constitution and data variance explained by each of the three main principal components.

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The potential of Pleurotus ostreatus spent mushroom compost (PSMC) as a green biosorbent for nickel (II) biosorption was investigated in this study. A novel approach of using the half-saturation concentration of biosorbent to rapidly determine the uptake, kinetics and mechanism of biosorption was employed together with cost per unit uptake analysis to determine the potential of this biosorbent. Fifty per cent nickel (II) biosorption was obtained at a half-saturation constant of 0.7 g biosorbent concentration, initial pH in the range of 4-8, 10 min contact time, 50 mL 50 mg/L nickel (II) initial concentration. The experimental data were well fitted with the Langmuir isotherm model and the maximum nickel (II) biosorption was 3.04 mg/g. The results corresponded well to a second pseudo order kinetic model with the coefficient of determination value of 0.9999. Based on FTIR analysis, the general alkyl, hydroxyl or amino, aliphatic alcohol and carbonyl functional groups of biosorbent were involved in the biosorption process. Therefore, biosorption of nickel (II) must involve several mechanisms simultaneously such as physical adsorption, chemisorption and ion exchange. Cost comparison for PSMC with Amberlite IRC-86 ion exchange resin indicates that the biosorbent has the potential to be developed into a cost effective and environmentally friendly treatment system.

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Pollutants such as human pharmaceuticals and synthetic hormones that are not covered by environmental legislation have increasingly become important emerging aquatic contaminants. This paper reports the development of a sensitive and selective multi-residue method for simultaneous determination and quantification of 23 pharmaceuticals and synthetic hormones from different therapeutic classes in water samples. Target pharmaceuticals include anti-diabetic, antihypertensive, hypolipidemic agents, ß2-adrenergic receptor agonist, antihistamine, analgesic and sex hormones. The developed method is based on solid phase extraction (SPE) followed by instrumental analysis using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) with 30 min total run time. River water samples (150 mL) and (sewage treatment plant) STP effluents (100 mL) adjusted to pH 2, were loaded into MCX (3 cm(3), 60 mg) cartridge and eluted with four different reagents for maximum recovery. Quantification was achieved by using eight isotopically labeled internal standards (I.S.) that effectively correct for losses during sample preparation and matrix effects during LC-ESI-MS/MS analysis. Good recoveries higher than 70% were obtained for most of target analytes in all matrices. Method detection limit (MDL) ranged from 0.2 to 281 ng/L. The developed method was applied to determine the levels of target analytes in various samples, including river water and STP effluents. Among the tested emerging pollutants, chlorothiazide was found at the highest level, with concentrations reaching up to 865 ng/L in STP effluent, and 182 ng/L in river water.

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Acinetobacter haemolyticus, a Gram-negative aerobic locally isolated bacterium, immobilized on wood-husk showed the ability to detoxify Cr(VI) to Cr(III). Wood-husk, a natural cellulose-based support material, packed in an upward-flow column was used as support material for bacterial attachment. Around 97% of the Cr(VI) in wastewater containing 15 mg L(-1) of Cr(VI) was reduced at a flow rate of 8.0 mL min(-1). The wastewater containing Cr(VI) was added with liquid pineapple wastewater as nutrient source for the bacteria. Electron microscopic examinations of the wood-husk after 42 days of column operation showed gradual colonization of the wood-husk by bacterial biofilm. The use of 0.1% (v/v) formaldehyde as a disinfecting agent inhibited growth of bacteria present in the final wastewater discharge. This finding is important in view of the ethical code regarding possible introduction of exogenous bacterial species into the environment.

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Possible application of a locally isolated environmental isolate, Acinetobacter haemolyticus to remediate Cr(VI) contamination in water system was demonstrated. Cr(VI) reduction by A. haemolyticus seems to favour the lower concentrations (10-30 mg/L). However, incomplete Cr(VI) reduction occurred at 70-100 mg/L Cr(VI). Initial specific reduction rate increased with Cr(VI) concentrations. Cr(VI) reduction was not affected by 1 or 10 mM sodium azide (metabolic inhibitor), 10 mM of PO(4)3-, SO4(2-), SO(3)2-, NO3- or 30 mg/L of Pb(II), Zn(II), Cd(II) ions. However, heat treatment caused significant dropped in Cr(VI) reduction to less than 20% only. A. haemolyticus cells loses its shape and size after exposure to 10 and 50 mg Cr(VI)/L as revealed from TEM examination. The presence of electron-dense particles in the cytoplasmic region of the bacteria suggested deposition of chromium in the cells.

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