The structural gene of elastase strain K (elastase from Pseudomonas aeruginosa strain K), namely HindIII1500PstI, was successfully sequenced to contain 1497 bp. The amino acid sequence, deduced from the nucleotide sequence, revealed that the mature elastase consists of 301 amino acids, with a molecular mass of 33.1 kDa, and contains a conserved motif HEXXH, zinc ligands and residues involved in the catalysis of elastase strain K. The structural gene was successfully cloned to a shuttle vector, pUCP19, and transformed into Escherichia coli strains TOP10, KRX, JM109 and Tuner™ pLacI as well as P. aeruginosa strains PA01 (A.T.C.C. 47085) and S5, with detection of significant protein expression. Overexpression was detected from transformants KRX/pUCP19/HindIII1500PstI of E. coli and PA01/pUCP19/HindIII1500PstI of P. aeruginosa, with increases in elastolytic activity to 13.83- and 5.04-fold respectively relative to their controls. In addition, recombinant elastase strain K showed considerable stability towards numerous organic solvents such as methanol, ethanol, acetone, toluene, undecan-1-ol and n-dodecane, which typically pose a detrimental effect on enzymes; our finding provides further information to support the potential application of the enzyme in synthetic industries, particularly peptide synthesis.
Soil contamination by hydrocarbons, especially by used lubricating oil, is a growing problem in developing countries, which poses a serious threat to the environment. Phytoremediation of these contaminated soils offers environmental friendly and a cost effective method for their remediation. Hibiscus cannabinus was studied for the remediation of soil contaminated with 2.5 and 1% used lubricating oil and treated with organic wastes [banana skin (BS), brewery spent grain (BSG) and spent mushroom compost (SMC)] for a period of 90 days under natural conditions. Loss of 86.4 and 91.8% used lubricating oil was recorded in soil contaminated with 2.5 and 1% oil and treated with organic wastes respectively at the end of 90 days. However, 52.5 and 58.9% oil loss was recorded in unamended soil contaminated with 2.5 and 1% oil, respectively. The plant did not accumulate hydrocarbon from the soil but shows appreciable accumulation of Fe and Zn in the root and stem of H. cannabinus at the end of the experiment. The first order kinetic rate of uptake of Fe and Zn in H. cannabinus was higher in organic wastes amendment treatments compared to the unamended treatments, which are extremely low. The results of this study suggest that H. cannabinus has a high potential for remediation of hydrocarbon and heavy metal contaminated soil.
The effects of thermochemical pretreatment and continuous thermophilic conditions on the composting of a mixture of rice straw residue and cattle manure were investigated using a laboratory-scale composting reactor. Results indicate that the composting period of rice straw can be shortened to less than 10 days by applying alkali pre-treatment and continuous thermophilic composting conditions. The parameters obtained on day 9 of this study are similar to the criteria level published by the Canadian Council of Ministers of the Environment. The moisture content, organic matter reduction, pH level, electrical conductivity, total organic carbon reduction, soluble chemical oxygen demand reduction, total Kjeldahl nitrogen, carbon-to-nitrogen ratio, and germination index were 62.07%, 16.99%, 7.30%, 1058 μS/cm, 17.00%, 83.43%, 2.06%, 16.75%, and 90.33%, respectively. The results of this study suggest that the application of chemical-biological integrated processes under thermophilic conditions is a novel method for the rapid degradation and maturation of rice straw residue.
The efficiency of the extraction and removal of pollutants from food and the environment has been an important issue in analytical science. By incorporating inorganic species into an organic matrix, a new material known as an organic-inorganic hybrid material is formed. As it possesses high selectivity, permeability, and mechanical and chemical stabilities, organic-inorganic hybrid materials constitute an emerging research field and have become popular to serve as sorbents in various separaton science methods. Here, we review recent significant advances in analytical solid-phase extraction employing organic-inorganic composite/nanocomposite sorbents for the extraction of organic and inorganic pollutants from various types of food and environmental matrices. The physicochemical characteristics, extraction properties, and analytical performances of sorbents are discussed; including morphology and surface characteristics, types of functional groups, interaction mechanism, selectivity and sensitivity, accuracy, and regeneration abilities. Organic-inorganic hybrid sorbents combined with extraction techniques are highly promising for sample preparation of various food and environmental matrixes with analytes at trace levels.
A series of tripeptide organocatalysts containing a secondary amine group and two amino acids with polar side chain units were developed and evaluated in the direct asymmetric intermolecular aldol reaction of 4-nitrobenzaldehyde and cyclohexanone. The effectiveness of short polar peptides as asymmetric catalysts in aldol reactions to attain high yields of enantio- and diastereoselective isomers were investigated. In a comparison, glutamic acid and histidine produced higher % ee and yields when they were applied as the second amino acid in short trimeric peptides. These short polar peptides were found to be efficient organocatalysts for the asymmetric aldol addition reaction in aqueous media.
Sonocatalytic degradation of various organic dyes (Congo Red, Reactive Blue 4, Methyl Orange, Rhodamine B and Methylene Blue) catalyzed by powder and nanotubes TiO(2) was studied. Both catalysts were characterized using transmission electron microscope (TEM), surface analyzer, Raman spectroscope and thermal gravimetric analyzer (TGA). Sonocatalytic activity of powder and nanotubes TiO(2) was elucidated based on the degradation of various organic dyes. The former catalyst was favorable for treatment of anionic dyes, while the latter was more beneficial for cationic dyes. Sonocatalytic activity of TiO(2) nanotubes could be up to four times as compared to TiO(2) powder under an ultrasonic power of 100 W and a frequency of 42 kHz. This was associated with the higher surface area and the electrostatic attraction between dye molecules and TiO(2) nanotubes. Fourier transform-infrared spectrometer (FT-IR) was used to identify changes that occurred on the functional group in Rhodamine B molecules and TiO(2) nanotubes after the reaction. Sonocatalytic degradation of Rhodamine B by TiO(2) nanotubes apparently followed the Langmuir-Hinshelwood adsorption kinetic model with surface reaction rate of 1.75 mg/L min. TiO(2) nanotubes were proven for their high potential to be applied in sonocatalytic degradation of organic dyes.
A field investigation was conducted at all water treatment plants throughout 11 states and Federal Territory in Peninsular Malaysia. The sampling points in this study include treatment plant operation, service reservoir outlet and auxiliary outlet point at the water pipelines. Analysis was performed by solid phase micro-extraction technique with a 100 microm polydimethylsiloxane fibre using gas chromatography with mass spectrometry detection to analyse 54 volatile organic compounds (VOCs) of different chemical families in drinking water. The concentration of VOCs ranged from undetectable to 230.2 microg/l. Among all of the VOCs species, chloroform has the highest concentration and was detected in all drinking water samples. Average concentrations of total trihalomethanes (THMs) were almost similar among all states which were in the range of 28.4--33.0 microg/l. Apart from THMs, other abundant compounds detected were cis and trans-1,2-dichloroethylene, trichloroethylene, 1,2-dibromoethane, benzene, toluene, ethylbenzene, chlorobenzene, 1,4-dichlorobenzene and 1,2-dichloro - benzene. Principal component analysis (PCA) with the aid of varimax rotation, and parallel factor analysis (PARAFAC) method were used to statistically verify the correlation between VOCs and the source of pollution. The multivariate analysis pointed out that the maintenance of auxiliary pipelines in the distribution systems is vital as it can become significant point source pollution to Malaysian drinking water.
In this study, tannin-based natural coagulant was used to treat stabilized landfill leachate. Tannin modified with amino group was utilized for the treatment process. Central composite design (CCD) was used to investigate and optimize the effect of tannin dosage and pH on four responses. The treatment efficiency was evaluated based on the removal of four selected (responses) parameters; namely, chemical oxygen demand (COD), color, NH3-N and total suspended solids (TSS). The optimum removal efficiency for COD, TSS, NH3-N and color was obtained using a tannin dosage of 0.73 g at a pH of 6. Moreover, the removal efficiency for selected heavy metals from leachate; namely, iron (Fe2+), zinc (Zn2+), copper (Cu2+), chromium (Cr2+), cadmium (Cd2+), lead (Pb2+), arsenic (As3+), and cobalt (Co2+) was also investigated. The results for removal efficiency for COD, TSS, NH3-N, and color were 53.50%, 60.26%, and 91.39%, respectively. The removal of selected heavy metals from leachate for Fe2+, Zn2+, Cu2+, Cr2+, Cd2+, Pb2+, As3+ and cobalt Co2+ were 89.76%, 94.61%, 94.15%, 89.94%, 17.26%, 93.78%, 86.43% and 84.19%, respectively. The results demonstrate that tannin-based natural coagulant could effectively remove organic compounds and heavy metals from stabilized landfill leachate.
LOD and LOQ are two important performance characteristics in method validation. This work compares three methods based on the International Conference on Harmonization and EURACHEM guidelines, namely, signal-to-noise, blank determination, and linear regression, to estimate the LOD and LOQ for volatile organic compounds (VOCs) by experimental methodology using GC. Five VOCs, toluene, ethylbenzene, isopropylbenzene, n-propylbenzene, and styrene, were chosen for the experimental study. The results indicated that the estimated LODs and LOQs were not equivalent and could vary by a factor of 5 to 6 for the different methods. It is, therefore, essential to have a clearly described procedure for estimating the LOD and LOQ during method validation to allow interlaboratory comparisons.
Nipah virus is a zoonotic virus isolated from an outbreak in Malaysia in 1998. The virus causes infections in humans, pigs, and several other domestic animals. It has also been isolated from fruit bats. The pathogenesis of Nipah virus infection is still not well described. In the present study, Nipah virus replication kinetics were estimated from infection of African green monkey kidney cells (Vero) using the one-step SYBR Green I-based quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR) assay.
Mature landfill leachate is typically non-biodegradable and contains high concentration of refractory organics. The aim of this research was to optimize operating parameters in electro-Fenton process, for the removal of recalcitrant organics from semi-aerobic landfill leachate using response surface methodology (RSM). Effectiveness of important process parameters H(2)O(2)/Fe(2+) molar ratio, current density, pH and reaction time were determined, optimized and modeled successfully. Significant quadratic polynomial models were obtained (R(2)=0.9972 and 0.9984 for COD and color removals, respectively). Numerical optimization based on desirability function were employed; in a 43 min trial 94.07% of COD and 95.83% of color were removed at pH 3 and H(2)O(2)/Fe(2+) molar ratio 1, while current density was 49 mA/cm(2). The results indicate that E-Fenton process was an effective technology for semi-aerobic landfill leachate treatment.
The main objective of this paper is to find the optimum operating condition to upgrade the EFB-derived pyrolysis oil (bio-oil) to liquid fuel, mainly gasoline using Taguchi Method. From the analysis that has been done, it is found that the optimum operating condition for heterogeneous catalytic cracking process is at 400 degrees C, 15min of reaction time using 30g of catalyst weight where operating at this condition produced the highest yield of gasoline fraction which is 91.67 wt.%. This observation proves that EFB-derived pyrolysis oil could be upgraded via heterogeneous catalytic cracking to produce gasoline.
The current available molecular method to detect infectious bursal disease virus (IBDV) is by reverse transcriptase-polymerase chain reaction (RT-PCR). However, the conventional PCR is time consuming, prone to error and less sensitive. In this study, the performances of Sybr Green I real-time PCR, enzyme-linked immunosorbent assay (ELISA) and conventional agarose detection methods in detecting specific IBDV PCR products were compared. We found the real-time PCR was at least 10 times more sensitive than ELISA detection method with a detection limit of 0.25pg. The latter was also at least 10 times more sensitive than agarose gel electrophoresis detection method. The developed assay detects both very virulent and vaccine strains of IBDV but not other RNA viruses such as Newcastle disease virus and infectious bronchitis virus. Hence, Sybr Green I-based real-time PCR is a highly sensitive assay for the detection of IBDV.
Organic photovoltaics (OPVs) die due to their interactions with environmental gases, i.e., moisture and oxygen, the latter being the most dangerous, especially under illumination, due to the fact that most of the active layers used in OPVs are extremely sensitive to oxygen. In this work we demonstrate solution-based effective barrier coatings based on composite of poly(vinyl butyral) (PVB) and mica flakes for the protection of poly (3-hexylthiophene) (P3HT)-based organic solar cells (OSCs) against photobleaching under illumination conditions. In the first step we developed a protective layer with cost effective and environmentally friendly methods and optimized its properties in terms of transparency, barrier improvement factor, and bendability. The developed protective layer maintained a high transparency in the visible region and improved oxygen and moisture barrier quality by the factor of ~7. The resultant protective layers showed ultra-flexibility, as no significant degradation in protective characteristics were observed after 10 K bending cycles. In the second step, a PVB/mica composite layer was applied on top of the P3HT film and subjected to photo-degradation. The P3HT films coated with PVB/mica composite showed improved stability under constant light irradiation and exhibited a loss of <20% of the initial optical density over the period of 150 h. Finally, optimized barrier layers were used as encapsulation for organic solar cell (OSC) devices. The lifetime results confirmed that the stability of the OSCs was extended from few hours to over 240 h in a sun test (65 °C, ambient RH%) which corresponds to an enhanced lifetime by a factor of 9 compared to devices encapsulated with pristine PVB.
Employment of edible oils as alternative green fuel for vehicles had raised debates on the sustainability of food supply especially in the third-world countries. The non-edible oil obtained from the abundantly available rubber seeds could mitigate this issue and at the same time reduce the environmental impact. Therefore, this paper investigates the catalytic cracking reaction of a model compound named linoleic acid that is enormously present in the rubber seed oil. Batch-scale experiments were conducted using 8.8 mL Inconel batch reactor having a cyclic horizontal swing span of 2 cm with a frequency of 60 cycles per minute at 450 °C under atmospheric condition for 90 min. The performance of HZSM-5, HBeta, HFerrierite, HMordenite and HY catalysts was tested for their efficiency in favouring gasoline range hydrocarbons. The compounds present in the organic liquid product were then analysed using GC-MS and classified based on PIONA which stands for paraffin, isoparaffin, olefin, naphthenes and aromatics respectively. The results obtained show that HZSM-5 catalyst favoured gasoline range hydrocarbons that were rich in aromatics compounds and promoted the production of desired isoparaffin. It also gave a higher cracking activity; however, large gaseous as by-products were produced at the same time.
The sources, distribution, transformation, toxicity and accumulation of persistent organic pollutants (POPs) in aquatic and terrestrial ecosystems have attracted global concern and attention over the last several decades. Although, POPs are toxic, degrade slowly and have a tendency to accumulate in the food chain, they are still widely used worldwide in many fields, such as industrial and agricultural activities. In addition, discharge of POPs into waterways may lead to serious health-related and environmental problems. This review provides an overview of the continental distributions of many types of POPs and the health risks associated with the exposure to POPs in daily life. This review also discusses the distribution of POPs in Malaysia, and the future work that will be conducted in the Klang River, one of the basins subjected to pollution due to development and urbanization.
The analysis of total organic carbon (TOC) by the American Public Health Association (APHA) closed-tube reflux colorimetric method requires potassium dichromate (K2Cr2O7), silver sulfate (AgSO4), and mercury (HgSO4) sulfate in addition to large volumes of both reagents and samples. The method relies on the release of oxygen from dichromate on heating which is consumed by carbon associated with organic compounds. The method risks environmental pollution by discharging large amounts of chromium (VI) and silver and mercury sulfates. The present method used potassium monochromate (K2CrO4) to generate the K2Cr2O7 on demand in the first phase. In addition, miniaturizing the procedure to semi microanalysis decreased the consumption of reagents and samples. In the second phase, mercury sulfate was eliminated as part of the digestion mixture through the introduction of sodium bismuthate (NaBiO3) for the removal of chlorides from the sample. The modified method, the potassium monochromate closed-tube colorimetry with sodium bismuthate chloride removal (KMCC-Bi), generates the potassium dichromate on demand and eliminates mercury sulfate. The semi microanalysis procedure leads to a 60% reduction in sample volume and ≈ 33.33 and 60% reduction in monochromate and silver sulfate consumption respectively. The LOD and LOQ were 10.17 and 33.90 mg L-1 for APHA, and 4.95 and 16.95 mg L-1 for KMCC-Bi. Recovery was between 83 to 98% APHA and 92 to 104% KMCC-Bi, while the RSD (%) ranged between 0.8 to 5.0% APHA and 0.00 to 0.62% KMCC-Bi. The method was applied for the UV-Vis spectrometry determination of COD in water and wastewater. Statistics was done by MINITAB 17 or MS Excel 2016. ᅟ Graphical abstract.
A new cell line, Asian sea bass brain (ASBB), was derived from the brain tissue of Asian sea bass Lates calcarifer. This cell line was maintained in Leibovitz L-15 media supplemented with 10% fetal bovine serum (FBS). The ASBB cell line was subcultured more than 60 times over a period of 15 mo. The ASBB cell line consists predominantly of fibroblastic-like cells and was able to grow at temperatures between 20°C and 30°C with an optimum temperature of 25°C. The growth rate of these cells increased as the proportion of FBS increased from 2% to 20% at 25°C with optimum growth at the concentrations of 10% or 15% FBS. Polymerase chain reaction products were obtained from ASBB cells and tissues of sea bass with primer sets of microsatellite markers of sea bass. An isolate of piscine nodavirus from juveniles of marine fish species tested positive by IQ2000 kit for viral nervous necrosis detection and was examined for its infectivity to a fish cell line of ASBB. A marine fish betanodavirus was tested to determine the susceptibility of this new cell line in comparison with commercial highly permissive SSN-1 cells. The ASBB cell line was found to be susceptible to nodavirus (RGNNV genotype), and the infection was confirmed by comparison cytopathic effect (CPE) with commercial SSN-1 and reverse transcriptase-polymerase chain reaction. A nodavirus was further elucidated by electron microscopy, and the virus tested was shown to induce CPE on ASBB cells with significant high titer. This suggests that the ASBB cell line has good potential for the isolation of fish viruses.
A novel adsorbent, magnetic, macro-reticulated cross-linked chitosan (MRC) was synthesised for the removal of tetracycline (TC) from water using a source of biogenic waste (gastropod shells) as a pore-forming agent. The insertion of crosslinks into the chitosan frame was confirmed by FTIR analysis, while the stability of the MRC was demonstrated via a stability test performed in an acidic solution. The enhanced porosity of the MRC was confirmed by the evaluation of its porosity, a swelling test and the determination of its specific surface area. The time-concentration profile of the sorption of TC onto the MRC demonstrated that equilibrium was attained relatively quickly (120 min), and the data obtained fitted a pseudo second order (r(2)>0.99) kinetic equation better than a pseudo first order or reversible first order kinetic equation. The optimisation of process variables indicated that the sorption of TC onto the MRC was favoured at a low solution pH and that the presence of organics (simulated by the addition of humic acid) negatively impacted the magnitude of TC removal. The area of coverage of TC on the MRC (2.51 m(2)/g) was low compared to the specific surface area of the MRC (47.95 m(2)/g). The value of the calculated energy of adsorption of TC onto the MRC was 100 kJ/mol, which is far above the range of 1-16 kJ/mol stipulated for physical adsorption.
The coagulation-flocculation process incorporated with membrane separation technology will become a new approach for palm oil mill effluent (POME) treatment as well as water reclamation and reuse. In our current research, a membrane pilot plant has been used for POME treatment where the coagulation-flocculation process plays an important role as a pretreatment process for the mitigation of membrane fouling problems. The pretreated POME with low turbidity values and high water recovery are the main objectives to be achieved through the coagulation-flocculation process. Therefore, treatment optimization to serve these purposes was performed using jar tests and applying a response surface methodology (RSM) to the results. A 2(3) full-factorial central composite design (CCD) was chosen to explain the effect and interaction of three factors: coagulant dosage, flocculent dosage, and pH. The CCD is successfully demonstrated to efficiently determine the optimized parameters, where 78% of water recovery with a 20 NTU turbidity value can be obtained at the optimum value of coagulant dosage, flocculent dosage, and pH at 15 000 mg/L, 300 mg/L, and 6, respectively.