Natural clays have recently been proven to possess antibacterial properties. Effective natural antimicrobial agents are needed to combat bacterial contamination on food contact surfaces, which are increasingly more prevalent in the food chain. This study sought to determine the antibacterial activity of clays against the food-borne pathogens Salmonella typhimurium ATCC 14028 and Staphylococcus aureus ATCC 13565. Soils were processed to yield leachates and suspensions from untreated and treated clays. Soil particle size, pH, cation-exchange capacity, metal composition and mineralogy were characterized. Antibacterial screening was performed on six Malaysian soils via the disc diffusion method. In addition, a time-kill assay was conducted on selected antibacterial clays after 6 h of exposure. The screening revealed that Munchong and Carey clays significantly inhibit Salmonella typhimurium (11.00 ± 0.71 mm) and S. aureus (7.63 ± 0.48 mm), respectively. Treated Carey clay leachate and suspension completely kill Salmonella typhimurium, while S. aureus viability is reduced (2 to 3 log10). The untreated Carey and all Munchong clays proved ineffective as antibacterials. XRD analysis confirmed the presence of pyrite and magnetite. Treated Carey clays had a higher soluble metal content compared to Munchong; namely Al (92.63 ± 2.18 mg/L), Fe (65.69 ± 3.09 mg/L) and Mg (88.48 ± 2.29 mg/L). Our results suggest that metal ion toxicity is responsible for the antibacterial activity of these clays.
An electrochemical microbiosensor for DNA has been fabricated based on new acrylic microspheres modified with reactive N-acryloxysuccinimide (NAS) functional groups. Hydrophobic poly(n-butylacrylate-N-acryloxysuccinimide) microspheres were synthesized in an emulsion form with a simple one-step photopolymerization technique. Aminated DNA probe was attached to the succinimde functional group of the acrylic microspheres via covalent bonding. The hybridization of the immobilized DNA probe with the complementary DNA was studied by differential pulse voltametry using anthraquninone-2-sulfonic acid monohydrate sodium salt (AQMS) as the electroactive hybridization label. The influences of many factors such as duration of DNA probe immobilization and hybridization, pH, type of ions, buffer concentrations, ionic strength, operational temperature and non-complementary DNA on the biosensor performance were evaluated. Under optimized conditions, the DNA microbiosensor demonstrated a linear response range to target DNA over a wide concentration range of 1.0 × 10(-16) and 1.0 × 10(-8) M with a lower limit of detection (LOD) of 9.46 × 10(-17) M (R(2) = 0.97). This DNA microbiosensor showed good reproducibility with 2.84% RSD (relative standard deviation) (n = 3). Application of the NAS-modified acrylic microspheres in the construction of DNA microbiosensor had improved the overall analytical performance of the resultant DNA microbiosensor when compared with other reported DNA biosensors using other nano-materials for membranes and microspheres as DNA immobilization matrices.
Bestari Jaya, former tin mining catchment covers an area of 2656.31 hectares comprised of four hundred and forty-two different-size lakes and ponds. The present study area comprise of 92 hectares of the catchment that include four large size lakes. Arc GIS version 9.2 used to develop bathymetric map, Global Positioning System (GPS) for hydrographical survey and flow meter was utilized for water discharge analysis (flow routing) of the catchment. The water quality parameters (pH, temperature, electric conductivity, dissolved oxygen DO, total dissolved solids TDS, chlorides, ammonium, nitrates) were analyzed by using Hydrolab. Quality assurance (QA) and quality control (QC) procedures were strictly followed throughout the field work and data analysis. Different procedures were employed to evaluate the analytical data and to check for possible transcription or dilution errors, changes during analysis, or unusual or unlikely values. The results obtained are compared with interim national water quality standards for Malaysia indicates that water quality of area is highly degraded. It is concluded that Bestri Jaya ex-mining catchment has a high pollution potential due to mining activities and River Ayer Hitam, recipient of catchment water, is a highly polluted river.
Colibacillosis is one of the main causes of economic loss in the poultry industry worldwide. Although antibiotics have been used to control this infection, the emergence of antibiotic-resistant bacteria poses a threat to animal and human health. Phage therapy has been reported as one of the potential alternative methods to control bacterial infections. However, efficient phage therapy is highly dependent on the characteristics of the phage isolated. In the present study the characteristics of a lytic phage, ØEC1, which was found to be effective against the causative agent of colibacillosis in chickens in a previous in vivo study, are reported.
A hollow fiber liquid phase microextraction (HF-LPME) in conjunction with reversed phase HPLC-UV method was developed for the extraction and determination of trace amounts of the antidiabetic drug, mitiglinide (MIT) in biological fluids. The drug was extracted from 10 mL aqueous sample (donor phase (DP)) into an organic phase impregnated in the pores of hollow fiber, followed by the back extraction into a second aqueous solution (acceptor phase (AP)) located in the lumen of the hollow fiber. Parameters influencing the extraction efficiency including the kind of organic solvent, composition of DP and AP, extraction time, stirring rate and salt addition were investigated and optimized. Under the optimized extraction conditions, high enrichment factors (210-fold), good linearity (5-1000 ng mL(-1)) and detection limit lower than 1.38 ng mL(-1) were achieved. Recoveries of spiked samples were in the range (88.3-96.3%) and (92.0-99.3%) for urine and plasma samples, respectively. The percent relative standard deviation (n=9) for the extraction and determination of three concentration levels (100, 400 and 800 ng mL(-1)) of MIT were less than 10.6% and 13.6% for urine and plasma samples, respectively. The developed method is simple, sensitive and has been successfully applied to the analysis of MIT in biological fluids.
Budu is a famous Malaysian fish sauce, usually used as seasoning and condiment in cooking. Budu is produced by mixing fish and salt at certain ratio followed by fermentation for six months in closed tanks. In this study, four commercial brands of Budu were analyzed for their chemical properties (pH, salt content and volatile compounds). The pH of Budu samples ranged from 4.50-4.92, while the salt (NaCl) content ranged between 11.80% and 22.50% (w/v). For tentative identification of volatile flavor compounds in Budu, two GC columns have been used, DB-WAX and HP-5MS. A total of 44 volatile compounds have been detected and 16 were common for both columns. 3-Methyl-1-butanol, 2-methylbutanal, 3-methylbutanal, dimethyl disulfide, 3-(methylthio)-propanal, 3-methylbutanoic acid and benzaldehye have been identified as the aroma-active compounds in Budu due to their lower threshold values.
The use of ice cubes in beverages is common among patrons of food outlets in Malaysia although its safety for human consumption remains unclear. Hence, this study was designed to determine the presence of faecal coliforms and several useful water physicochemical parameters viz. free residual chlorine concentration, turbidity and pH in ice cubes from 30 randomly selected food outlets in Kubang Kerian, Kelantan. Faecal coliforms were found in ice cubes in 16 (53%) food outlets ranging between 1 CFU/100mL to >50 CFU/ 100mL, while in the remaining 14 (47%) food outlets, in samples of tap water as well as in commercially bottled drinking water, faecal coliforms were not detected. The highest faecal coliform counts of >50 CFU/100mL were observed in 3 (10%) food outlets followed by 11-50 CFU/100mL and 1-10 CFU/100mL in 7 (23%) and 6 (20%) food outlets, respectively. All samples recorded low free residual chlorine concentration (<0.10mg/L) with the pH ranging between 5.5 and 7.3 and turbidity between 0.14-1.76 NTU. Since contamination by faecal coliforms was not detected in 47% of the samples, tap water and commercially bottled drinking water, it was concluded that (1) contamination by faecal coliforms may occur due to improper handling of ice cubes at the food outlets or (2) they may not be the water sources used for making ice cubes. Since low free residual chlorine concentrations were observed (<0.10mg/ L) in all samples as well as in both tap water and commercially bottled drinking water, with the pH ranged between 5.5-7.3, ineffective disinfection of water source as a contributing factor to such high counts of faecal coliforms in ice cubes also could not be ruled out. Therefore, a periodical, yet comprehensive check on the food outlets, including that of ice cube is crucial in ensuring better food and water for human consumption.
A cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) technique has been developed for enantioseparation of vinpocetine using an inexpensive 2-hydroxypropyl-β-CD (HP-β-CD) as the chiral selector (CS). The best chiral separation was achieved using 40 mM HP-β-CD as the CS in 50 mM phosphate buffer (pH 7.0) consisting of 40 mM sodium dodecyl sulfate (SDS) at a separation temperature and separation voltage of 25°C and 25 kV, respectively. To the author's best knowledge, this is the first CD-MEKC study able to successfully separate the four stereoisomer of vinpocetine in separation time of 9.5 min and resolution of 1.04-3.87.
Two genes that encode α-amylases from two Anoxybacillus species were cloned and expressed in Escherichia coli. The genes are 1,518 bp long and encode 506 amino acids. Both sequences are 98% similar but are distinct from other well-known α-amylases. Both of the recombinant enzymes, ASKA and ADTA, were purified using an α-CD-Sepharose column. They exhibited an optimum activity at 60°C and pH 8. Both amylases were stable at pH 6-10. At 60°C in the absence of Ca²⁺, negligible reduction in activity for up to 48 h was observed. The activity half-life at 65°C was 48 and 3 h for ASKA and ADTA, respectively. In the presence of Ca²⁺ ions, both amylases were highly stable for at least 48 h and had less than a 10% decrease in activity at 70°C. Both enzymes exhibited similar end-product profiles, and the predominant yield was maltose (69%) from starch hydrolysis. To the best of our knowledge, most α-amylases that produce high levels of maltose are active at an acidic to neutral pH. This is the first report of two thermostable, alkalitolerant recombinant α-amylases from Anoxybacillus that produce high levels of maltose and have an atypical protein sequence compared with known α-amylases.
The effective removal of o-cresol is currently both an environmental and economic challenge. ZnO is not only an efficient photocatalyst but is also cost effective, as its photoabsorption can extend from the ultraviolet (UV) to the visible range thereby allowing the use of inexpensive visible light sources, such as sunlight. The principal objective of the present work is to investigate the visible light-driven removal of o-cresol from aqueous solution in the presence of 1.0 wt% Mn-doped ZnO. To measure the efficiency ofphotodegradation, the variables studied included the amount ofphotocatalyst, concentration of o-cresol, pH and irradiation time. The concentration ofo-cresol and residual organic carbon was monitored using a UV-visible spectrophotometer, ultra high-pressure liquid chromatography and a total organic carbon analyser. The optimum conditions under which the photodegradation of o-cresol was most favourable corresponded to 1.5 g/l ZnO, 35 ppm o-cresol and pH 9. The ZnO-1 wt% Mn photoprocess has demonstrated reusability for more than three times, which warrants its scale-up from laboratory- to in industrial-scale application.
An in-vial liquid-liquid microextraction method was developed for the selective extraction of the phenolic acids (caffeic, gallic, cinnamic, ferulic, chlorogenic, syringic, vanillic, benzoic, p-hydroxybenzoic, 2,4-dihydroxybenzoic, o-coumaric, m-coumaric and p-coumaric) in vegetable oil samples. The optimised extraction conditions for 20 g sample were: volume of diluent (n-hexane), 2 mL; extractant, methanol: 5 mM sodium hydroxide (60:40; v/v); volume of extractant, 300 μL (twice); vortex, 1 min; centrifugation, 5 min. Recoveries for the studied phenolic acids were 80.1-119.5%. The simultaneous determination of the phenolic acid extracts was investigated by capillary electrophoresis (CE). Separations were carried out on a bare fused-silica capillary (50 μm i.d.× 40 cm length) involving 25 mM sodium tetraborate (pH 9.15) and 5% methanol as CE background electrolyte in the normal polarity mode, voltage of 30 kV, temperature of 25°C, injection time of 4s (50 mbar) and electropherograms were recorded at 200 nm. The phenolic acids were successfully separated in less than 10 min. The validated in-vial LLME-CE method was applied to the determination of phenolic acids in vegetable oil samples (extra virgin olive oil, virgin olive oil, pure olive oil, walnut oil and grapeseed oil). The developed method shows significant advantages over the current methods as lengthy evaporation step is not required.
Protease is one of the most important industrial enzymes with a multitude of applications in both food and non-food sectors. Although most commercial proteases are microbial proteases, the potential of non-conventional protease sources, especially plants, should not be overlooked. In this study, horse mango (Mangifera foetida Lour) fruit, known to produce latex with a blistering effect upon contact with human skin, was chosen as a source of protease, and the effect of the extraction process on its protease activity evaluated. The crude enzyme was extracted from the kernels and extraction was optimized by a response surface methodology (RSM) using a central composite rotatable design (CCRD). The variables studied were pH (x(1)), CaCl(2) (x(2)), Triton X-100 (x(3)), and 1,4-dithryeitol (x(4)). The results obtained indicate that the quadratic model is significant for all the variables tested. Based on the RSM model generated, optimal extraction conditions were obtained at pH 6.0, 8.16 mM CaCl(2), 5.0% Triton X-100, and 10.0 mM DTT, and the estimated response was 95.5% (w/w). Verification test results showed that the difference between the calculated and the experimental protease activity value was only 2%. Based on the t-value, the effects of the variables arranged in ascending order of strength were CaCl(2) < pH < DTT < Triton X-100.
The behavioral responses of guppy Poecilia reticulata (Poeciliidae) and prawn Macrobrachium lanchesteri (Palaemonidae) individuals exposed to acid mine drainage (AMD) were monitored online in the laboratory with a Multispecies Freshwater Biomonitor™ (MFB). These responses were compared to those to reference water acidified to the respective pH values (ACID). Test animals in the juvenile stage were used for both species and were exposed to AMD and ACID for 24 hours. The stress behaviors of both test animals consisted mainly of decreased activity in AMD and increased activity in ACID, indicating that the metals in the AMD played a role as a stress factor in addition to pH. The locomotor activity levels of guppies and prawns for the ACID treatment were higher than the locomotor activity levels for the AMD treatment with increasing pH value. For guppies, significant differences were observed when specimens were exposed to AMD and ACID at pH 5.0 and 6.0; the percentage activities were only 16% and 12%, respectively, for AMD treatment, whereas for ACID treatment, the percentage activities were 35% and 40%, respectively, similar to the value of 36% for the controls. Similar trends were also observed for prawns, for which the percentage activities were only 6% and 4%, respectively, for AMD treatment, whereas for ACID treatment, the percentage activities were 31% and 38%, respectively, compared to 44% in the controls. This study showed that both species are suitable for use as indicators for ecotoxicity testing with the MFB.
In this article, modified κ-carrageenan hydrogel nanocomposites were synthesized to increase the release ability of carrageenan hydrogels under gastrointestinal conditions. The effect of MgO nanoparticle loading in a model drug (methylene blue) release is investigated. Characterization of hydrogels were carried out using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), Field Emission Scanning Electron Microscope (FESEM) and Differential Scanning Calorimetry (DSC). Genipin was used to increase the delivery performance in gastrointestinal tract delivery by decreasing release in simulated stomach conditions and increasing release in simulated intestine conditions. It is shown that the amount of methylene blue released from genipin-cross-linked nanocomposites can be 67.5% higher in intestine medium and 56% lower in the stomach compared to κ-carrageenan hydrogel. It was found that by changing the nanoparticle loading and genipin concentration in the composite, the amount of drug released can be monitored. Therefore, applying nanoparticles appears to be a potential strategy to develop controlled drug delivery especially in gastrointestinal tract studies.
Mango peel is a good source of protease but remains an industrial waste. This study focuses on the optimization of polyethylene glycol (PEG)/dextran-based aqueous two-phase system (ATPS) to purify serine protease from mango peel. The activity of serine protease in different phase systems was studied and then the possible relationship between the purification variables, namely polyethylene glycol molecular weight (PEG, 4000-12,000 g·mol(-1)), tie line length (-3.42-35.27%), NaCl (-2.5-11.5%) and pH (4.5-10.5) on the enzymatic properties of purified enzyme was investigated. The most significant effect of PEG was on the efficiency of serine protease purification. Also, there was a significant increase in the partition coefficient with the addition of 4.5% of NaCl to the system. This could be due to the high hydrophobicity of serine protease compared to protein contaminates. The optimum conditions to achieve high partition coefficient (84.2) purification factor (14.37) and yield (97.3%) of serine protease were obtained in the presence of 8000 g·mol(-1) of PEG, 17.2% of tie line length and 4.5% of NaCl at pH 7.5. The enzymatic properties of purified serine protease using PEG/dextran ATPS showed that the enzyme could be purified at a high purification factor and yield with easy scale-up and fast processing.
The synthesis of oxygenated fuel additives via solvent freebase-catalyzed etherification of glycerol is reported. The products of glycerol etherification arediglycerol (DG) and triglycerol (TG) with DG being the favorable one. The catalytic activity of different homogeneous alkali catalysts (LiOH, NaOH, KOH and Na(2)CO(3)) was investigated during the glycerol etherification process. LiOH exhibited an excellent catalytic activity during this reaction, indicated by the complete glycerol conversion with a corresponding selectivity of 33% toward DG. The best reaction conditions were a reaction temperature of 240°C, a catalyst/glycerol mass ratio of 0.02 and a reaction time of 6h. The influences of various reaction variables such as nature of the catalyst, catalyst loading, reaction time and reaction temperature on glycerol etherification were elucidated. Industrially, the findings attained in this study might contribute towards promoting the biodiesel industry through utilization of its by-products.
Biosorption potential of mustard oil cake (MOC) for Ni(II) from aqueous medium was studied. Spectroscopic studies showed possible involvement of acidic (hydroxyl, carbonyl and carboxyl) groups in biosorption. Optimum biosorption was observed at pH 8. Contact time, reaction temperature, biosorbent dose and adsorbate concentration showed significant influence. Linear and non-linear isotherms comparison suggests applicability of Temkin model at 303 and 313 K and Freundlich model at 323K. Kinetics studies revealed applicability of Pseudo-second-order model. The process was endothermic and spontaneous. Freundlich constant (n) and activation energy (Ea) values confirm physical nature of the process. The breakthrough and exhaustive capacities for 5 mg/L initial Ni(II) concentration were 0.25 and 4.5 mg/g, while for 10 mg/L initial Ni(II) concentration were 4.5 and 9.5 mg/g, respectively. Batch desorption studies showed maximum Ni(II) recovery in acidic medium. Regeneration studies by batch and column process confirmed reutilization of biomass without appreciable loss in biosorption.
A high concentration of phosphorus in wastewater may lead to excessive algae growth and deoxygenation of the water. In this work, nanofiltration (NF) of phosphorus-rich solutions is studied in order to investigate its potential in removing and recycling phosphorus. Wastewater samples from a pulp and paper plant were first analyzed. Commercial membranes (DK5, MPF34, NF90, NF270, NF200) were characterized and tested in permeability and phosphorus removal experiments. NF90 membranes offer the highest rejection of phosphorus; a rejection of more than 70% phosphorus was achieved for a feed containing 2.5 g/L of phosphorus at a pH <2. Additionally, NF90, NF200 and NF270 membranes show higher permeability than DK5 and MPF34 membranes. The separation performance of NF90 is slightly affected by phosphorus concentration and pressure, which may be due to concentration polarization and fouling. By adjusting the pH to 2 or adding sulfuric acid, the separation performance of NF90 was improved in removing phosphorus. However, the presence of acetic acid significantly impairs the rejection of phosphorus.
Preparation of activated carbon has been attempted using KOH as activating agent by microwave heating from biodiesel industry solid residue, oil palm empty fruit bunch (EFBAC). The significance of chemical impregnation ratio (IR), microwave power and activation time on the properties of activated carbon were investigated. The optimum condition has been identified at the IR of 1.0, microwave power of 600 W and activation time of 7 min. EFBAC was characterized by scanning electron microscopy, Fourier transform infrared spectroscopy and nitrogen adsorption isotherm. The surface chemistry was examined by zeta potential measurement, determination of surface acidity/basicity, while the adsorptive property was quantified using methylene blue as dye model compound. The optimum conditions resulted in activated carbon with a monolayer adsorption capacity of 395.30 mg/g and carbon yield of 73.78%, while the BET surface area and total pore volume were corresponding to 1372 m2/g and 0.76 cm3/g, respectively.
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.