Contamination of toxic metals in P. viridis mussels has been prevalently reported; hence, health risk assessment for consuming this aquaculture product as well as the surrounding surface seawater at its harvesting sites appears relevant. Since Kampung Pasir Puteh, Pasir Gudang is the major harvesting site in Malaysia, and because the last heavy metal assessment was done in 2009, its current status remains unclear. Herein, flame atomic absorption spectrometry and flow injection mercury/hydride system were used to determine the concentrations of Pb, Cd, Cu and total Hg in P. viridis mussels and surface seawater (January-March 2015), respectively. Significantly higher concentrations of these metals were found in P. viridis mussels (p
The escalating trend of murder victim concealment worldwide appears worrying, and literature does not reveal any specific study focusing on victim concealment amongst convicted male Malaysian murderers. Therefore, this study was aimed at investigating the psychological traits that may underlie the act of murder concealment in Malaysia via mixed method approaches. Male murderers (n = 71) from 11 prisons were selected via purposive sampling technique. In the quantitative analysis, a cross-sectional study design using the validated questionnaire was used. The questionnaire contained murder concealment variables and four Malay validated psychometric instruments measuring: personality traits, self-control, aggression, and cognitive distortion. The independent sample t-tests revealed the significantly higher level of anger in murderers who did not commit concealment acts (8.55 ± 2.85, p < 0.05) when compared with those who did so (6.40 ± 2.64). Meanwhile, the Kruskal-Wallis H test revealed that anger and the personality trait of aggressiveness-hostility significantly varied across the different groups of murder concealment acts (p < 0.05). The qualitative data obtained via the in-depth interviews revealed two important themes for the murderers to commit murder concealment acts: (1) fear of discovery and punishment and (2) blaming others. These findings discussed from the perspectives of the murderers within the context of criminology and psychology may provide the first ever insight into the murder concealment acts in Malaysia that can benefit the relevant authorities for crime prevention and investigation efforts.
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.
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.
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.
In this study, nanocellulose (NC) was successfully extracted from oil palm frond leaves (OPFL) using a combination of bleaching, alkaline treatment and acid hydrolysis. X-ray diffractogram revealed the extracted NC was crystalline with a crystallinity index of 70.2%. This indicates its suitability as nano-fillers for preparing the chitosan/nanocellulose (CS-NC) supports to immobilize Candida rugosa lipase (CRL) to produce the CRL/CS-NC biocatalysts. FTIR, FESEM and TGA characterizations of the CRL/CS-NC confirm the CRLs were successfully conjugated to the CS-NC supports. The air-dried CS-NC supports gave satisfactory immobilization of the CRLs (5.2mg/g) with the resultant CRL/CS-NCs catalysed conversions of ≥80% of butyl butyrate within 6h. Time course reaction profile revealed that 76.3% butyl butyrate conversion was achieved at 4h immobilization time using 3mg/mL of CRL/CS-NCs. NMR analyses on the purified butyl butyrate confirmed that the ester was successfully synthesized.
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.
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.
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.
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.
The discovery of forensic evidence (e.g. weapons) during forensic underwater investigations has seen an increasing trend. To date, small particle reagent (SPR) has been one of the routinely used methods for visualising fingerprints on wet, non-porous substrates. However, the long term use of SPR is detrimental to humans and environment due to the use of toxic chemicals. Although previously we have successfully developed and optimised a greener nanobio-based reagent (NBR), its suitable practical use in a more realistic scene (e.g. outdoor pond) was not evaluated. Therefore, this present research is aimed at (1) investigating the performance of NBR against the benchmark SPR in visualising fingerprints immersed in a natural outdoor pond and (2) evaluating the greenness of NBR against the analytical Eco-Scale. Results showed that the performance of the optimised NBR was mostly comparable (University of Canberra (UC) comparative scale: 0) with SPR at visualising fingerprints on three different non-porous substrates immersed in a natural outdoor pond. Observably, the NBR had higher preference towards aged fingerprints (up to 4 weeks of immersion). In addition, its greenness assessment revealed 76 points, indicating 'excellent green analysis'. The findings gathered here further supported the practical use of the NBR in forensic investigations.
The use of 21 autosomal STR loci for human identification has been gaining popularity throughout the world. It has been indicated that the forensic statistical parameters for supporting the use of 21 STR loci varied among different populations. Hitherto, such data for the diverse Malaysian populations remain unreported, rendering doubts in the court of law about its real ability for human identification in Malaysian population. Using the GlobalFiler™ Express PCR Amplification Kit, complete DNA profiles of 21 STR loci from buccal swabs of convicted Malaysian criminal (n = 570; 190 each for Malays, Chinese, and Indians) (by the year 2016-2017) were analyzed for their allele frequencies, exact test of Hardy-Weinberg equilibrium, observed and expected heterozygosity, power of discrimination, power of exclusion, match probability, and polymorphism information content. Most of the loci were found to be in the Hardy-Weinberg equilibrium after the Bonferroni correction. Being the most informative locus, SE33 demonstrated the highest power of discrimination and power of exclusion, indicating its usefulness to discriminate individuals. In contrast, TPOX had the lowest power of discrimination and power of exclusion, as well as being the less informative genetic locus for all Malaysian population studied here. The probabilities that two individuals would share the same DNA profiles among the Malaysian Malays, Chinese, and Indians, as well as in general Malaysian population, were 1.3713 × 10-25, 2.8822 × 10-25, 7.5668 × 10-26, and 1.0385 × 10-26, respectively. The results obtained here were found comparable with similar studies reported in other populations. Hence, its robustness for forensic human identification among the Malaysian populations is, therefore, statistically supported.
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.
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.
Waterways are popular locations for the disposition of criminal evidence because the recovery of latent fingerprints from such evidence is difficult. Currently, small particle reagent is a method often used to visualize latent fingerprints containing carcinogenic and hazardous compounds. This study proposes an eco-friendly, safranin-tinted Candida rugosa lipase (triacylglycerol ester hydrolysis EC 3.1.1.3) with functionalized carbon nanotubes (CRL-MWCNTS/GA/SAF) as an alternative reagent to the small particle reagent. The CRL-MWCNTS/GA/SAF reagent was compared with the small particle reagent to visualize groomed, full fingerprints deposited on stainless steel knives which were immersed in a natural outdoor pond for 30 days. The quality of visualized fingerprints using the new reagent was similar (modified-Centre for Applied Science and Technology grade: 4; p > 0.05) to small particle reagent, even after 15 days of immersion. Despite the slight decrease in quality of visualized fingerprints using the CRL-MWCNTS/GA/SAF on the last three immersion periods, the fingerprints remained forensically identifiable (modified-Centre for Applied Science and Technology grade: 3). The possible chemical interactions that enabled successful visualization is also discussed. Thus, this novel reagent may provide a relatively greener alternative for the visualization of latent fingerprints on immersed non-porous objects.
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.
The onus of proof in criminal cases is beyond any reasonable doubt, and the issue on the lack of complete internal validation data can be manipulated when it comes to justifying the validity and reliability of the X-chromosomal short tandem repeats analysis for court representation. Therefore, this research evaluated the efficiency of the optimized 60% reduced volumes for polymerase chain reaction (PCR) amplification using the Qiagen Investigator® Argus X-12 QS Kit, as well as the capillary electrophoresis (CE) sample preparation for blood samples on Flinder's Technology Associates (FTA) cards. Good-quality DNA profile (3000-12,000 RFU) from the purified blood sample on FTA card (1.2 mm) were obtained using the optimized PCR (10.0 μL of PCR reaction volume and 21 cycles) and CE (9.0 μL Hi-Di™ Formamide and 0.3 μL DNA Size Standard 550 [BTO] and 27 s injection time) conditions. The analytical and stochastic thresholds were 100 and 200 RFU, respectively. Hence, the internal validation data supported the use of the optimized 60% reduced PCR amplification reaction volume of the Qiagen Investigator® Argus X-12 QS Kit as well as the CE sample preparation for producing reliable DNA profiles that comply with the quality assurance standards for forensic DNA testing laboratories, while optimizing the analytical cost.
Being commonly found at crime scenes, fingerprints are crucial for human identification, attributable to their uniqueness, persistence and systematic classification of ridge patterns. In addition to latent fingerprints being invisible to the naked eye, the escalating trends of disposing forensic evidence bearing such prints in watery bodies would further complicate criminal investigations. Taking into account the toxicity of small particle reagent (SPR) commonly used in visualising latent fingerprints on wet and non-porous objects, a greener alternative using the nanobio-based reagent (NBR) has been suggested. However, NBR only applies to white and/or relatively light-coloured objects. Thus, conjugation of sodium fluorescein dye with NBR (f-NBR) may be beneficial for increasing the contrast of fingerprint on multi-colored objects. Hence, this study was aimed at investigating the possibility of such conjugation (i.e., f-NBR) as well as proposing suitable interactions between the f-NBR and lipid constituents of fingerprints (tetra-, hexa- and octadecanoic acids) via molecular docking and molecular dynamics simulations. The binding energies between CRL with its ligands were observed at -8.1, -5.0, -4.9 and -3.6 kcal/mole for sodium fluorescein, tetra-, hexa- and octadecanoic acids, respectively. Besides, the formations of hydrogen bonds observed in all complexes (ranged between 2.6 and 3.4 Å), further supported by the stabilized root mean square deviation (RMSDs) plots in MD simulations. In short, the conjugation of f-NBR was computationally feasible, and thereby merits further investigations in the laboratory.Communicated by Ramaswamy H. Sarma.
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.
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.