Displaying publications 1 - 20 of 35 in total

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  1. Influence of paraquat on Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) infesting minced-beef substrates in Kelantan, Malaysia
    Mahat NA, Yin CL, Jayaprakash PT
    J Forensic Sci, 2014 Mar;59(2):529-32.
    PMID: 24745083
    This study investigated the influence of paraquat, a prevalent poison used by suicides, on initial oviposition and development of Chrysomya megacephala (Fabricius) using minced-beef substrates. Paraquat in lethal dose for human (40 mg/kg), two times the lethal dose (80 mg/kg) and five times the lethal dose (200 mg/kg) were mixed thoroughly with respective minced-beef substrates (1 kg each) that were decomposed in a shaded habitat fully protected from rain. Results of four replications of the above experiment revealed that the presence of paraquat neither delayed initial oviposition nor prolonged the developmental stages of C. megacephala. Therefore, estimation of postmortem interval (PMI) based on empirical baseline data obtained using animal models devoid of any poisons would still be appropriate for estimating PMI in paraquat-related deaths.
  2. Influence of rain and malathion on the oviposition and development of blowflies (Diptera: Calliphoridae) infesting rabbit carcasses in Kelantan, Malaysia
    Mahat NA, Zafarina Z, Jayaprakash PT
    Forensic Sci Int, 2009 Nov 20;192(1-3):19-28.
    PMID: 19671490 DOI: 10.1016/j.forsciint.2009.07.008
    The influence of rain and malathion on the initial oviposition as well as development of blowfly species infesting rabbit carcasses decomposing in sunlit and shaded habitats were studied over a period of 1 year in Kelantan, Malaysia. Chrysomya megacephala (Fabricius) was the most dominant species that infested the carcasses, followed by Chrysomya rufifacies (Macquart). In general, rain, depending on its intensity, delayed initial oviposition by 1-2 days and prolonged the pupation period by 1-3 days. The presence of malathion in the carcasses delayed initial oviposition by 1-3 days and prolonged the pupation period by 2-3 days. These findings deserve consideration while estimating postmortem interval since rain is a commonplace occurrence in Malaysia and malathion is one of the common poisons as an agent for choice to commit suicide.
  3. Ternary biogenic silica/magnetite/graphene oxide composite for the hyperactivation of Candida rugosa lipase in the esterification production of ethyl valerate
    Jacob AG, Wahab RA, Mahat NA
    Enzyme Microb Technol, 2021 Aug;148:109807.
    PMID: 34116744 DOI: 10.1016/j.enzmictec.2021.109807
    Oil palm leaves (OPL) silica (SiO2) can replace the energy-intensive, commercially produced SiO2. Moreover, the agronomically sourced biogenic SiO2 is more biocompatible and cost-effective enzyme support, which properties could be improved by the addition of magnetite (Fe3O4) and graphene oxide (GO) to yield better ternary support to immobilize enzymes, i.e., Candida rugosa lipase (CRL). This study aimed to optimize the Candida rugosa lipase (CRL immobilization onto the ternary OPL-silica-magnetite (Fe3O4)-GO (SiO2/Fe3O4/GO) support, for use as biocatalyst for ethyl valerate (EV) production. Notably, this is the first study detailing the CRL/SiO2/Fe3O4/GO biocatalyst preparation for rapid and high yield production of ethyl valerate (EV). AFM and FESEM micrographs revealed globules of CRL covalently bound to GL-A-SiO2/Fe3O4/GO; similar to Raman and UV-spectroscopy results. FTIR spectra revealed amide bonds at 3478 cm-1 and 1640 cm-1 from covalent interactions between CRL and GL-A-SiO2/Fe3O4/GO. Optimum immobilization conditions were 4% (v/v) glutaraldehyde, 8 mg/mL CRL, at 16 h stirring in 150 mM NaCl at 30 °C, offering 24.78 ± 0.26 mg/g protein (specific activity = 65.24 ± 0.88 U/g). The CRL/SiO2/Fe3O4/GO yielded 77.43 ± 1.04 % of EV compared to free CRL (48.75 ± 0.70 %), verifying the suitability of SiO2/Fe3O4/GO to hyperactivate and stabilize CRL for satisfactory EV production.
  4. Fulltext Presence of faecal coliforms and selected heavy metals in ice cubes from food outlets in Taman Universiti, Johor Bahru, Malaysia
    Mahat NA, Meor Ahmad Z, Abdul Wahab R
    Trop Biomed, 2015 Sep;32(3):471-7.
    PMID: 26695207 MyJurnal
    Consumption of iced beverages is common in Malaysia although specific research focusing on its safety parameters such as presence of faecal coliforms and heavy metal elements remains scarce. A study conducted in Kelantan indicated that faecal coliforms were detected in the majority of the ice cube samples analyzed, largely attributable to improper handling. Hence, it was found pertinent to conduct similar study in other parts of the country such as Johor Bahru if the similar pattern prevailed. Therefore, this present cross sectional study which randomly sampled ice cubes from 30 permanent food outlets in Taman Universiti, Johor Bahru for detecting contamination by faecal coliforms and selected heavy metal elements (lead, copper, manganese and zinc) acquires significance. Faecal coliforms were detected in 11 (36.67%) of the samples, ranging between 1 CFU/100 mL to > 50 CFU/100 mL; two of the samples were grossly contaminated (>50 CFU/100 mL). Interestingly, while positive detection of lead was observed in 29 of the 30 ice cube samples (mean: 0.511±0.105 ppm; range: 0.489-0.674 ppm), copper, manganese and zinc were not detected. In addition, analysis on commercially bottled mineral water as well as in tap water samples did not detect such contaminations. Therefore, it appears that (1) contamination of faecal coliforms in ice cubes in food outlets in Malaysia may not be sporadic in pattern but rather prevalent and (2) the source of water used for manufacturing the ice cubes that contained significant amount of lead would suggest that (3) it was neither originated from the treated tap water supply nor bottled mineral water or (4) perhaps contaminated during manufacturing process. Further studies exploring the source of water used for manufacturing these ice cubes as well as the handling process among food operators deserve consideration.
  5. Fulltext Malathion extraction from larvae of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) for determining death due to malathion
    Mahat NA, Jayaprakash PT, Zafarina Z
    Trop Biomed, 2012 Mar;29(1):9-17.
    PMID: 22543598 MyJurnal
    The use of Chrysomya megacephala larvae for detecting malathion for diagnosing the cause of death was investigated. This could prove useful when the visceral organs have become liquefied during decomposition and therefore cannot be sampled. A field experiment was conducted in which C. megacephala were allowed to colonise naturally the corpses of rabbits that had died of malathion poisoning. The concentration of malathion increased gradually during the larval stages of C. megacephala reaching the maximum concentration in the third instar larvae. The concentration of malathion declined during prepupal stage and reached its lowest level among tenerals. The average malathion concentrations in C. megacephala growing in poisoned rabbit corpses left in a sunlit habitat were significantly higher (p<0.05) than those growing on poisoned rabbits left in a shaded habitat. The concentrations of malathion in the different stages of development of C. megacephala were moderately correlated (r = 0.51-0.69) with the administered doses as well as with those estimated in visceral organs. Thus, it would not be reliable to suggest the formulation of mathematical algorithms for relating the concentration of malathion found in the different stages of development of C. megacephala with those found in the visceral organs. However, in the context of forensic investigation, the qualitative detection of malathion in C. megacephala may prove useful in diagnosing the cause of death, since malathion is a common cause of accidental and suicidal deaths.
  6. Biophysical characterization of a recombinant lipase KV1 from Acinetobacter haemolyticus in relation to pH and temperature
    Batumalaie K, Khalili E, Mahat NA, Huyop F, Wahab RA
    Biochimie, 2018 Sep;152:198-210.
    PMID: 30036604 DOI: 10.1016/j.biochi.2018.07.011
    Spectroscopic and calorimetric methods were employed to assess the stability and the folding aspect of a novel recombinant alkaline-stable lipase KV1 from Acinetobacter haemolyticus under varying pH and temperature. Data on far ultraviolet-circular dichroism of recombinant lipase KV1 under two alkaline conditions (pH 8.0 and 12.0) at 40 °C reveal strong negative ellipticities at 208, 217, 222 nm, implying its secondary structure belonging to a α + β class with 47.3 and 39.0% ellipticity, respectively. Results demonstrate that lipase KV1 adopts its most stable conformation at pH 8.0 and 40 °C. Conversely, the protein assumes a random coil structure at pH 4.0 and 80 °C, evident from a strong negative peak at ∼ 200 nm. This blue shift suggests a general decline in enzyme activity in conjunction with the partially or fully unfolded state that invariably exposed more hydrophobic surfaces of the lipase protein. The maximum emission at ∼335 nm for pH 8.0 and 40 °C indicates the adoption of a favorable protein conformation with a high number of buried tryptophan residues, reducing solvent exposure. Appearance of an intense Amide I absorption band at pH 8.0 corroborates an intact secondary structure. A lower enthalpy value for pH 4.0 over pH 8.0 and 12.0 in the differential scanning calorimetric data corroborates the stability of the lipase at alkaline conditions, while a low Km (0.68 ± 0.03 mM) for tributyrin verifies the high affinity of lipase KV1 for the substrate. The data, herein offer useful insights into future structure-based tunable catalytic activity of lipase KV1.
  7. Survival of antibiotic resistant Escherichia coli in vacuumpacked keropok lekor: Food safety alert among SME keropok lekor producers
    Wan-Hamat H, Lani MN, Hamzah Y, Alias R, Hassan Z, Mahat NA
    Trop Biomed, 2020 Mar 01;37(1):103-115.
    PMID: 33612722
    The microbiological quality of thirty ready-to-eat (RTE) keropok lekor (a sausage shape Malaysian fish product) was evaluated in comparison to microbiological guidelines for ready to eat foods. The two E. coli isolates were subjected to DNA sequencing, identified and tested for their resistance towards fifteen different antibiotics. The survival and growth of the isolated E. coli strains inoculated in keropok lekor at atmospheric air and vacuum packaging were also evaluated. Results revealed that four samples (13.33%) contained Enterobacteriaceae counts that exceeded the recommended allowable counts of 4.0 log10 CFU/g. Unsatisfactory level of coliforms (< 1.7 log10 CFU/g) was also observed in ten of the samples; two of which contained E. coli (2.1 ± 0.17 and 3.7 ± 0.02 log10 CFU/g), suggesting of poor hygiene and sanitation practices. While the 'Possible E10' E. coli strain was observably resistant towards Nalidixic acid (30µg) alone, B10 E. coli isolate was worryingly resistant towards Ampicillin (10µg), Ceftazidime (30µg), Ciprofloxacin (5µg), Ceftriaxone (30µg), Nalidixic acid (30µg) and Tetracycline (30µg). This study also revealed that the growth and survival of the 'Possible E10' and B10 E. coli strains were not significantly affected by vacuum packaging when stored at both 4°C and 28°C. Therefore, intervention programmes to alert and educate smallmedium enterprisers (SMEs) of keropok lekor producers on food safety as well as potential health risks that can be associated due to inappropriate handling procedures of such product, merits consideration.
  8. Occurrence of heavy metals and their removal in Perna viridis mussels using chemical methods: a review
    Ali TG, Abdul Keyon AS, Mahat NA
    Environ Sci Pollut Res Int, 2022 Jan;29(4):4803-4821.
    PMID: 34775561 DOI: 10.1007/s11356-021-17343-3
    Despite the nutritional benefits, bivalves like mussels are also an excellent aquatic heavy metal biomonitoring agent due to their high tolerance to varying levels of temperature, salinity and oxygen, as well as pollutants. Although the accumulated toxic heavy metals may not exert direct negative impacts on the mussels, such toxicants in mussel tissues can give harmful effects on human body when consumed in toxic quantities and/or over prolonged period. The booming of urban and industrial activities, and consequently the increment of runoffs, as well as wastewater effluents and leaching, further exacerbated the magnitude of this issue. Hence, continuous monitoring of heavy metal contents in mussels is vital to ensure its compliance with food safety regulations, protecting consumers at large. This review paper discusses the occurrence of heavy metals in mussels especially that of Perna viridis in Malaysia and other parts of the world since year 2000 until 2021. Heavy metal concentration data and patterns from various coastal and/or estuaries were compared. Where applicable, statistical data that indicate variations between sampling sites, sampling months or years and chemical treatments for heavy metal removal were critically reviewed. Health risk assessment findings were also discussed. More importantly, related chemical-based interventions to minimize and/or eliminate toxic heavy metals from mussels are also reviewed.
  9. Synthesis of geranyl propionate in a solvent-free medium using Rhizomucor miehei lipase covalently immobilized on chitosan-graphene oxide beads
    Isah AA, Mahat NA, Jamalis J, Attan N, Zakaria II, Huyop F, et al.
    Prep Biochem Biotechnol, 2017 Feb 07;47(2):199-210.
    PMID: 27341522 DOI: 10.1080/10826068.2016.1201681
    The chemical route of producing geranyl propionate involves the use of toxic chemicals, liberation of unwanted by-products as well as problematic separation process. In view of such problems, the use of Rhizomucor miehei lipase (RML) covalently bound onto activated chitosan-graphene oxide (RML-CS/GO) support is suggested. Following analyses using Fourier transform infrared spectroscopy, field emission scanning electron microscopy, transmission electron microscopy, and thermogravimetry, properties of the RML-CS/GO were characterized. A response surface methodological approach using a 3-level-four-factor (incubation time, temperature, substrate molar ratio, and stirring rate) Box-Behnken design was used to optimize the experimental conditions to maximize the yield of geranyl propionate. Results revealed that 76 ± 0.02% of recovered protein had yielded 7.2 ± 0.04 mg g(-1) and 211 ± 0.3% U g(-1) of the maximum protein loading and esterification activity, respectively. The actual yield of geranyl propionate (49.46%) closely agreed with the predicted value (49.97%) under optimum reaction conditions (temperature: 37.67°C, incubation time: 10.20 hr, molar ratio (propionic acid:geraniol): 1:3.28, and stirring rate: 100.70 rpm) and hence, verifying the suitability of this approach. Since the method is performed under mild conditions, the RML-CS/GO biocatalyst may prove to be an environmentally benign alternative for producing satisfactory yield of geranyl propionate.
  10. Patterns of oviposition and development of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) and Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) on burned rabbit carcasses
    Mahat NA, Zainol-Abidin NL, Nordin NH, Abdul-Wahab R, Jayaprakash PT
    Forensic Sci Int, 2016 Mar;260:9-13.
    PMID: 26779962 DOI: 10.1016/j.forsciint.2015.12.047
    Considering that crimes against animals such as illegal killing and cruelty have been alarmingly increasing and since burning is one of the common ways for disposing cadavers, ability to estimate minimum postmortem interval (PMI) using entomological data merits consideration. Chrysomya megacephala and Chrysomya rufifacies are common necrophagous species recovered from cadavers in many countries including Malaysia. Specific studies focusing on the oviposition and developmental patterns of both species on cadavers manifesting different levels of burn as described by the Crow-Glassman Scale (CGS) remain scarce. In four replicates, rabbit carcasses were burned to CGS levels #1, #2 and #3 by varying the amount of petrol used and duration of burning. Oviposition by C. megacephala and C. rufifacies was delayed by one day in the case of carcasses burned to the CGS level #3 (p<0.05) when compared with that of controls. Such delay in oviposition was not observed in the CGS level #1 and #2 carcasses. No significant differences (p>0.05) in the duration of development were found between control and burned carcasses. These findings deserve consideration while estimating minimum PMI since burning as a mean for disposing animal and human cadavers is gaining popularity.
  11. Candida rugosa Lipase Immobilized onto Acid-Functionalized Multi-walled Carbon Nanotubes for Sustainable Production of Methyl Oleate
    Che Marzuki NH, Mahat NA, Huyop F, Buang NA, Wahab RA
    Appl Biochem Biotechnol, 2015 Oct;177(4):967-84.
    PMID: 26267406 DOI: 10.1007/s12010-015-1791-z
    The chemical production of methyl oleate using chemically synthesized fatty acid alcohols and other toxic chemicals may lead to significant environmental hazards to mankind. Being a highly valuable fatty acid replacement raw material in oleochemical industry, the mass production of methyl oleate via environmentally favorable processes is of concern. In this context, an alternative technique utilizing Candida rugosa lipase (CRL) physically adsorbed on multi-walled carbon nanotubes (MWCNTs) has been suggested. In this study, the acid-functionalized MWCNTs prepared using a mixture of HNO3 and H2SO4 (1:3 v/v) was used as support for immobilizing CRL onto MWCNTs (CRL-MWCNTs) as biocatalysts. Enzymatic esterification was performed and the efficiency of CRL-MWCNTs was evaluated against the free CRL under varying conditions, viz. temperature, molar ratio of acid/alcohol, solvent log P, and enzyme loading. The CRL-MWCNTs resulted in 30-110 % improvement in the production of methyl oleate over the free CRL. The CRL-MWCNTs attained its highest yield (84.17 %) at 50 °C, molar ratio of acid/alcohol of 1:3, 3 mg/mL of enzyme loading, and iso-octane (log P 4.5) as solvent. Consequently, physical adsorption of CRL onto acid-functionalized MWCNTs has improved the activity and stability of CRL and hence provides an environmentally friendly means for the production of methyl oleate.
  12. Structure and properties of lipase activated by cellulose-silica polyethersulfone membrane for production of pentyl valerate
    Elias N, Wahab RA, Chandren S, Jamalis J, Mahat NA, Jye LW
    Carbohydr Polym, 2020 Oct 01;245:116549.
    PMID: 32718641 DOI: 10.1016/j.carbpol.2020.116549
    Herein, this study extracted nanocrystalline cellulose (NC) and silica (SiO2) from raw oil palm leaves (OPL), and employed as nanofillers in polyethersulfone (PES) to produce NC-SiO2-PES as support to immobilize Candida rugosa lipase (CRL) (NC-SiO2-PES/CRL). XRD, TGA-DTG and FTIR-ATR data affirmed that NC and SiO2 were isolated from OPL with corresponding crystallinity indices of 68 % and 70 %. A 0.02 cm membrane size with 5% (w/v) of NC-SiO2 without PVP K30 was optimal for membrane fabrication. CRL immobilized on the Glut-AP-NC-SiO2-PES membrane gave a higher conversion of pentyl valerate (PeVa) (91.3 %, p < 0.05) compared to Glut-NC-SiO2-PES (73.9 %) (p < 0.05). Characterization of the NC-SiO2-PES/CRL biocatalyst verified the presence of CRL. Hence, raw OPL is a proven good source of NC and SiO2, as reinforcement nanofillers in PES. The overall findings envisage the promising use of NC-SiO2-PES/CRL to catalyze an expedient and high yield of PeVa, alongside the suitability of NC-SiO2-PES for activating other enzymes.
  13. Fulltext Antioxidant Activity Evaluation of FlexirubinType Pigment from Chryseobacterium artocarpi CECT 8497 and Related Docking Study
    Mogadem A, Almamary MA, Mahat NA, Jemon K, Ahmad WA, Ali I
    Molecules, 2021 Feb 12;26(4).
    PMID: 33673263 DOI: 10.3390/molecules26040979
    The current research is focused on studying the biological efficacy of flexirubin, a pigment extracted from Chryseobacterium artocarpi CECT 8497.Different methods such as DPPH, H2O2, NO•, O2•-, •OH, lipid peroxidation inhibition by FTC and TBA, ferric reducing and ferrous chelating activity were carried out to evaluate the antioxidant activity of flexirubin. Molecular docking was also carried out, seeking the molecular interactions of flexirubin and a standard antioxidant compound with SOD enzyme to figure out the possible flexirubin activity mechanism. The new findings revealed that the highest level of flexirubin exhibited similar antioxidant activity as that of the standard compound according to the H2O2, •OH, O2•-, FTC and TBA methods. On the other hand, flexirubin at the highest level has shown lower antioxidant activity than the positive control according to the DPPH and NO• and even much lower when measured by the FRAP method. Molecular docking showed that the interaction of flexirubin was in the binding cavity of the SOD enzyme and did not affect its metal-binding site. These results revealed that flexirubin has antioxidant properties and can be a useful therapeutic compound in preventing or treating free radical-related diseases.
  14. Insight into the Rhizomucor miehei lipase supported on chitosan-chitin nanowhiskers assisted esterification of eugenol to eugenyl benzoate
    Abd Manan FM, Attan N, Zakaria Z, Mahat NA, Abdul Wahab R
    J Biotechnol, 2018 May 28;280:19-30.
    PMID: 29852195 DOI: 10.1016/j.jbiotec.2018.05.015
    To overcome drawbacks in the conventional chemical route to synthesize eugenyl benzoate, immobilized Rhizomucor miehei lipase (RML) as the biocatalyst was proposed. The RML conjugated to a hybrid support consisting of biopolymers, chitosan (CS) and chitin nanowhiskers (CNWs). 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDAC) was used as the crosslinker to bind the lipase. Immobilization of RML was the highest on crosslinked CS/CNWs which gave a protein loading of ∼8.12 mg/g, corresponding to specific and residual activity of 537 U/g and 137%, respectively. Fourier transform infrared spectroscopy, thermogravimetric analysis-differential thermogravimetry, field emission scanning electron and atomic force microscopy of RML-CS/CNWs revealed that RML was successfully attached to the surface of crosslinked CS/CNWs. Under an optimized condition, the highest yield of eugenyl benzoate (56.3%) was attained after 5 h using 3 mg/mL of RML-CS/CNWs with molar ratio of eugenol: benzoic acid of 3:1, as compared to only 47.3% for the free RML. Analyses of FTIR and NMR on purified eugenyl benzoate affirmed that the ester was successfully produced in the enzymatic esterification. Therefore, the use of the RML-CS/CNWs biocatalysts appears promising to afford good yields of eugenyl benzoate within a relatively shorter reaction time.
  15. A facile enzymatic synthesis of geranyl propionate by physically adsorbed Candida rugosa lipase onto multi-walled carbon nanotubes
    Mohamad NR, Buang NA, Mahat NA, Lok YY, Huyop F, Aboul-Enein HY, et al.
    Enzyme Microb Technol, 2015 May;72:49-55.
    PMID: 25837507 DOI: 10.1016/j.enzmictec.2015.02.007
    In view of several disadvantages as well as adverse effects associated with the use of chemical processes for producing esters, alternative techniques such as the utilization of enzymes on multi-walled carbon nanotubes (MWCNTs), have been suggested. In this study, the oxidative MWCNTs prepared using a mixture of HNO3 and H2SO4 (1:3 v/v) were used as a supportive material for the immobilization of Candida rugosa lipase (CRL) through physical adsorption process. The resulting CRL-MWCNTs biocatalysts were utilized for synthesizing geranyl propionate, an important ester for flavoring agent as well as in fragrances. Enzymatic esterification of geraniol with propionic acid was carried out using heptane as a solvent and the efficiency of CRL-MWCNTs as a biocatalyst was compared with the free CRL, considering the incubation time, temperature, molar ratio of acid:alcohol, presence of desiccant as well as its reusability. It was found that the CRL-MWCNTs resulted in a 2-fold improvement in the percentage of conversion of geranyl propionate when compared with the free CRL, demonstrating the highest yield of geranyl propionate at 6h at 55°C, molar ratio acid: alcohol of 1:5 and with the presence of 1.0g desiccant. It was evident that the CRL-MWCNTs biocatalyst could be reused for up to 6 times before a 50% reduction in catalytic efficiency was observed. Hence, it appears that the facile physical adsorption of CRL onto F-MWCNTs has improved the activity and stability of CRL as well as served as an alternative method for the synthesis of geranyl propionate.
  16. Fulltext Contamination of faecal coliforms in ice cubes sampled from food outlets in Kubang Kerian, Kelantan
    Noor Izani NJ, Zulaikha AR, Mohamad Noor MR, Amri MA, Mahat NA
    Trop Biomed, 2012 Mar;29(1):71-6.
    PMID: 22543605 MyJurnal
    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.
  17. Provenance Establishment of Stingless Bee Honey Using Multi-element Analysis in Combination with Chemometrics Techniques
    Shadan AF, Mahat NA, Wan Ibrahim WA, Ariffin Z, Ismail D
    J Forensic Sci, 2018 Jan;63(1):80-85.
    PMID: 28383137 DOI: 10.1111/1556-4029.13512
    As consumption of stingless bee honey has been gaining popularity in many countries including Malaysia, ability to identify accurately its geographical origin proves pertinent for investigating fraudulent activities for consumer protection. Because a chemical signature can be location-specific, multi-element distribution patterns may prove useful for provenancing such product. Using the inductively coupled-plasma optical emission spectrometer as well as principal component analysis (PCA) and linear discriminant analysis (LDA), the distributions of multi-elements in stingless bee honey collected at four different geographical locations (North, West, East, and South) in Johor, Malaysia, were investigated. While cross-validation using PCA demonstrated 87.0% correct classification rate, the same was improved (96.2%) with the use of LDA, indicating that discrimination was possible for the different geographical regions. Therefore, utilization of multi-element analysis coupled with chemometrics techniques for assigning the provenance of stingless bee honeys for forensic applications is supported.
  18. Statistical optimization and operational stability of Rhizomucor miehei lipase supported on magnetic chitosan/chitin nanoparticles for synthesis of pentyl valerate
    Rahman INA, Attan N, Mahat NA, Jamalis J, Abdul Keyon AS, Kurniawan C, et al.
    Int J Biol Macromol, 2018 Aug;115:680-695.
    PMID: 29698760 DOI: 10.1016/j.ijbiomac.2018.04.111
    The chemical-catalyzed transesterification process to produce biofuels i.e. pentyl valerate (PeVa) is environmentally unfriendly, energy-intensive with tedious downstream treatment. The present work reports the use of Rhizomucor miehei lipase (RML) crosslinked onto magnetic chitosan/chitin nanoparticles (RML-CS/CH/MNPs). The approach used to immobilize RML onto the CS/CH/MNPs yielded RML-CS/CH/MNPs with an immobilized protein loading and specific activity of 7.6 mg/g and 5.0 U·g-1, respectively. This was confirmed by assessing data of field emission scanning electron microscopy, X-ray diffraction, thermal gravimetric analysis and Fourier transform infrared spectroscopy. A three-level-four-factor Box-Behnken design (incubation time, temperature, substrate molar ratio, and enzyme loading) was used to optimize the RML-CS/CH/MNP-catalyzed esterification synthesis of PeVa. Under optimum condition, the maximum yield of PeVa (97.8%) can be achieved in 5 h at 50 °C using molar ratio valeric acid:pentanol (1:2) and an enzyme load of 2 mg/mL. Consequently, operational stability experiments showed that the protocol adopted to prepare the CS/CH/MNP nanoparticles had increased the durability of RML. The RML-CS/CH/MNP could catalyze up to eight successive esterification cycles to produce PeVa. The study also demonstrated the functionality of CS/CH/MNP nanoparticles as an eco-friendly support matrix for improving enzymatic activity and operational stability of RML to produce PeVa.
  19. Application of Box-Behnken design for ultrasound-assisted extraction and recycling preparative HPLC for isolation of anthraquinones from Cassia singueana
    Jibril S, Basar N, Sirat HM, Wahab RA, Mahat NA, Nahar L, et al.
    Phytochem Anal, 2019 Jan;30(1):101-109.
    PMID: 30288828 DOI: 10.1002/pca.2795
    INTRODUCTION: Cassia singueana Del. (Fabaceae) is a rare medicinal plant used in the traditional medicine preparations to treat various ailments. The root of C. singueana is a rich source of anthraquinones that possess anticancer, antibacterial and antifungal properties.

    OBJECTIVE: The objective of this study was to develop an ultrasound-assisted extraction (UAE) method for achieving a high extraction yield of anthraquinones using the response surface methodology (RSM), Box-Behnken design (BBD), and a recycling preparative high-performance liquid chromatography (HPLC) protocol for isolation of anthraquinones from C. singueana.

    METHODOLOGY: Optimisation of UAE was performed using the Box-Behnken experimental design. Recycling preparative HPLC was employed to isolate anthraquinones from the root extract of C. singueana.

    RESULTS: The BBD was well-described by a quadratic polynomial model (R2  = 0.9751). The predicted optimal UAE conditions for a high extraction yield were obtained at: extraction time 25.00 min, temperature 50°C and solvent-sample ratio of 10 mL/g. Under the predicted conditions, the experimental value (1.65 ± 0.07%) closely agreed to the predicted yield (1.64%). The obtained crude extract of C. singueana root was subsequently purified to afford eight anthraquinones.

    CONCLUSION: The extraction protocol described here is suitable for large-scale extraction of anthraquinones from plant extracts.

  20. Development patterns of necrophagous flies infesting rabbit carcasses decomposing in Mount Kapur Cave and its surrounding primary forest in Kuching, Sarawak
    Nordin NH, Ahmad UK, Abdul Rahim NA, Kamaluddin MR, Ismail D, Muda NW, et al.
    Trop Biomed, 2020 Jun 01;37(2):333-356.
    PMID: 33612803
    In addition to the scarcity of forensic entomology baseline data on oviposition of necrophagous insects and completion of their life cycles in the Borneo region, similar data derived from caves remain unreported. Since entomological baseline data can differ from one biogeoclimatic region to another, the lack of such data would limit the practical values of applying entomological evidence in estimating minimum postmortem interval (mPMI). Therefore, this present research that investigated oviposition and completion of life cycles of necrophagous flies infesting rabbit carcasses decomposing in Mount Kapur Cave and its surrounding forest habitat in Kuching, Sarawak merits forensic consideration. In general, 13 taxa of necrophagous flies were identified viz. Hypopygiopsis violacea, Hypopygiopsis fumipennis, Hemipyrellia ligurriens, Hemipyrellia tagaliana, Chrysomya megacephala, Chrysomya villeneuvi, Chrysomya rufifacies, Chrysomya chani, Chrysomya pinguis, Chrysomya nigripes, Ophyra spinigera and Ophyra chalcogaster, as well as unidentified Sarcophagidae. In addition, Hyp. violacea and Hyp. fumipennis were the two earlier necrophagous flies that oviposited in all rabbit carcasses decomposing in both habitats. While all these necrophagous flies were observed infesting carcasses in Mount Kapur Cave, Hem. ligurriens and Hem. tagaliana were not found infesting carcasses in the surrounding forest habitat. Complete life cycles for six and five different necrophagous fly species were successfully observed in Mount Kapur Cave and its surrounding forest habitat, respectively. Significant delay in oviposition, as well as longer durations for completing the life cycles in several necrophagous fly species were observed in Mount Kapur Cave when compared with those of surrounding forest habitat (p < 0.05). These findings deserve consideration as the first ever forensic empirical baseline data on oviposition and completion of life cycles for necrophagous flies in Sarawak as well as in a cave habitat, in view of its practical values for estimating mPMI for forensic practical caseworks.
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