The concentration profile, distribution and risk assessment of pharmaceutically active compounds (PhACs) in the coastal surface water from the Klang River estuary were measured. Surface coastal water samples were extracted using offline solid phase, applying polymeric C18 cartridges as extraction sorbent and measuring with liquid chromatography mass spectrometry-mass spectrometry (LC MS-MS) technique. Extraction method was optimized for its recovery, sensitivity and linearity. Excellent recoveries were obtained from the optimized method with percentage of recoveries ranging from 73 to 126%. The optimized analytical method achieved good sensitivity with limit of detection ranging from 0.05 to 0.15 ng L-1, while linearity of targeted compounds in the LC MS-MS system was more than 0.990. The results showed that amoxicillin has the highest concentration (102.31 ng L-1) followed by diclofenac (10.80 ng L-1) and primidone (7.74 ng L-1). The percentage of contribution (% of total concentration) for the targeted PhACs is in the following order; amoxicillin (92.90%) > diclofenac (3.95%) > primidone (1.23%) > dexamethasone (0.75%) > testosterone (0.70%) > sulfamethoxazole (0.33%) > progesterone (0.14%). Environmental risk assessment calculated based on deterministic approach (the RQ method), showed no present risk from the presence of PhACs in the coastal water of Klang River estuary. Nonetheless, this baseline assessment can be used for better understanding on PhACs pollution profile and distribution in the tropical coastal and estuarine ecosystem as well as for future comparative studies.
This baseline assessment reports on the occurrence, distribution, and sources of emerging organic contaminants (EOCs) in tropical coastal sediments of anthropogenically impacted Klang River estuary, Malaysia. Bisphenol A was the highest concentration detected at 16.84 ng g-1 dry weight, followed by diclofenac (13.88 ng g-1 dry weight) and E1 (12.47 ng g-1 dry weight). Five compounds, namely, amoxicillin, progesterone, diazinon, bisphenol A, and E1, were found in all sampling stations assessed, and other compounds such as primidone, diclofenac, testosterone, E2, and EE2 were ubiquitously present in sediment samples, with percentage of detection range from 89.04% to 98.38%. Organic carbon content and pH were the important factors controlling the fate of targeted compounds in the tropical estuarine sediment. On the basis of the literature from other studies, the sources of EOCs are thought to be from wastewater treatment plants, domestic/medical waste discharge, livestock activities, industrial waste discharge, and agricultural activities.
There is no consensus regarding the classification of optic neuritis, and precise diagnostic criteria are not available. This reality means that the diagnosis of disorders that have optic neuritis as the first manifestation can be challenging. Accurate diagnosis of optic neuritis at presentation can facilitate the timely treatment of individuals with multiple sclerosis, neuromyelitis optica spectrum disorder, or myelin oligodendrocyte glycoprotein antibody-associated disease. Epidemiological data show that, cumulatively, optic neuritis is most frequently caused by many conditions other than multiple sclerosis. Worldwide, the cause and management of optic neuritis varies with geographical location, treatment availability, and ethnic background. We have developed diagnostic criteria for optic neuritis and a classification of optic neuritis subgroups. Our diagnostic criteria are based on clinical features that permit a diagnosis of possible optic neuritis; further paraclinical tests, utilising brain, orbital, and retinal imaging, together with antibody and other protein biomarker data, can lead to a diagnosis of definite optic neuritis. Paraclinical tests can also be applied retrospectively on stored samples and historical brain or retinal scans, which will be useful for future validation studies. Our criteria have the potential to reduce the risk of misdiagnosis, provide information on optic neuritis disease course that can guide future treatment trial design, and enable physicians to judge the likelihood of a need for long-term pharmacological management, which might differ according to optic neuritis subgroups.
The need to isolate efficient heavy metal reducers for cost effective bioremediation strategy have resulted in the isolation of a potent molybdenum-reducing bacterium. The isolate was tentatively identified as Serratia sp. strain DRY5 based on the Biolog GN carbon utilization profiles and partial 16S rDNA molecular phylogeny. Strain DRY5 produced 2.3 times the amount of Mo-blue than S. marcescens strain Dr.Y6, 23 times more than E. coli K12 and 7 times more than E. cloacae strain 48. Strain DRY5 required 37 degrees C and pH 7.0 for optimum molybdenum reduction. Carbon sources such as sucrose, maltose, glucose and glycerol, supported cellular growth and molybdate reduction after 24 hr of static incubation. The most optimum carbon source that supported reduction was sucrose at 1.0% (w/v). Ammonium sulphate, ammonium chloride, glutamic acid, cysteine, and valine supported growth and molybdate reduction with ammonium sulphate as the optimum nitrogen source at 0. 2% (w/v). Molybdate reduction was optimally supported by 30 mM molybdate. The optimum concentration of phosphate for molybdate reduction was 5 mM when molybdate concentration was fixed at 30 mM and molybdate reduction was totally inhibited at 100 mM phosphate. Mo-blue produced by this strain shows a unique characteristic absorption profile with a maximum peak at 865 nm and a shoulder at 700 nm, Dialysis tubing experiment showed that 95.42% of Mo-blue was found in the dialysis tubing suggesting that the molybdate reduction seen in this bacterium was catalyzed by enzyme(s). The characteristics of isolate DRY5 suggest that it would be useful in the bioremediation ofmolybdenum-containing waste.
This paper describes a short-amplicon-based TaqMan probe quantitative real-time PCR (qPCR) assay for the quantitative detection of canine meat in chicken nuggets, which are very popular across the world, including Malaysia. The assay targeted a 100-bp fragment of canine cytb gene using a canine-specific primer and TaqMan probe. Specificity against 10 different animals and plants species demonstrated threshold cycles (Ct) of 16.13 ± 0.12 to 16.25 ± 0.23 for canine DNA and negative results for the others in a 40-cycle reaction. The assay was tested for the quantification of up to 0.01% canine meat in deliberately spiked chicken nuggets with 99.7% PCR efficiency and 0.995 correlation coefficient. The analysis of the actual and qPCR predicted values showed a high recovery rate (from 87% ± 28% to 112% ± 19%) with a linear regression close to unity (R(2) = 0.999). Finally, samples of three halal-branded commercial chicken nuggets collected from different Malaysian outlets were screened for canine meat, but no contamination was demonstrated.
A polymerase chain reaction (PCR) assay for the assessment of dog meat adulteration in meatballs was developed. The assay selectively amplified a 100-bp region of canine mitochondrial cytochrome b gene from pure, raw, processed and mixed backgrounds. The specificity of the assay was tested against 11 animals and 3 plants species, commonly available for meatball formulation. The stability of the assay was proven under extensively autoclaving conditions that breakdown target DNA. A blind test from ready to eat chicken and beef meatballs showed that the assay can repeatedly detect 0.2% canine meat tissues under complex matrices using 0.04 ng of dog DNA extracted from differentially treated meatballs. The simplicity, stability and sensitivity of the assay suggested that it could be used in halal food industry for the authentication of canine derivatives in processed foods.
Porcine blood is potentially being utilized in food as a binder, gelling agent, emulsifier or colorant. However, for certain communities, the usage of animal blood in food is strictly prohibited owing to religious concerns and health reasons. This study reports the development of monoclonal antibodies (MAbs) against heat-treated soluble proteins (HSPs) of autoclaved porcine blood; characterization of MAbs against blood, non-blood and plasma from different animal species using qualitative indirect non-competitive enzyme-linked immunosorbent assay (ELISA); and immunoblotting of antigenic components in HSPs of porcine blood.
The grow out of Asian seabass Lates calcarifer in marine net-cages is a popular aquaculture activity in Malaysia. Production of this species is greatly affected by the occurrence of vibriosis, which causes heavy mortality. Generally, young fish are more susceptible; they exhibit anorexia and skin darkening, followed by heavy mortality. The acutely affected older fish may also exhibit bloody lesions around the anus and the base of the fins. Twenty-one bacterial isolates obtained from internal organs (kidney, heart, spleen and liver) of the affected specimens were subjected to phenotypic characterization, testing for antibiotic susceptibility, and 16S ribosomal DNA sequencing. The sequencing result showed that all of the bacterial isolates belonged to Vibrio harveyi. The phenotypic characterization, however, identified 4 of the bacterial isolates as V. harveyi, 16 as V. parahaemolyticus, and 1 as V. alginolyticus. These findings suggest that biochemical features alone cannot be reliably used to identify bacterial pathogens, including V. harveyi, in aquaculture. Antibiotic susceptibility assays showed that some antibiotics, including oxytetracycline, nitrofurantoin, furazolidone, streptomycin, sulfamethoxazole, chloramphenicol, nalidixic acid, and oxolinic acid were effective against V. harveyi. Considering the side effects of these antibiotics, however, their use is not recommended in the aquaculture of Asian seabass.
Currently, the authentication analysis of edible fats and oils is an emerging issue not only by producers but also by food industries, regulators, and consumers. The adulteration of high quality and expensive edible fats and oils as well as food products containing fats and oils with lower ones are typically motivated by economic reasons. Some analytical methods have been used for authentication analysis of food products, but some of them are complex in sampling preparation and involving sophisticated instruments. Therefore, simple and reliable methods are proposed and developed for these authentication purposes. This review highlighted the comprehensive reports on the application of infrared spectroscopy combined with chemometrics for authentication of fats and oils. New findings of this review included (1) FTIR spectroscopy combined with chemometrics, which has been used to authenticate fats and oils; (2) due to as fingerprint analytical tools, FTIR spectra have emerged as the most reported analytical techniques applied for authentication analysis of fats and oils; (3) the use of chemometrics as analytical data treatment is a must to extract the information from FTIR spectra to be understandable data. Next, the combination of FTIR spectroscopy with chemometrics must be proposed, developed, and standardized for authentication and assuring the quality of fats and oils.
Brown rice is unpolished rice with immeasurable benefits for human health. Brown rice (BR) and pre-germinated brown rice (PGBR) are known to contain various functional compounds such as gamma-oryzanol, dietary fibre and gamma-aminobutyric acid (GABA). In the present study, the experimental diets containing BR and PGBR (24, 48 h pre-germination) were used to investigate the influence of pre-germination time of brown rice on blood cholesterol in Sprague-Dawley male rats.
The effect of transesterification of engkabang (Shorea macrophylla) fat - canola oil (EF35/ CaO65) blend by Mucor miehei lipase (1%, w/w oil) in a solvent-free system was investigated at different time intervals of 6 h, 12 h and 24 h. Compositional changes of the samples withdrawn at specified time intervals while the reaction in progress were analysed by chromatography, whereas the polymorphic forms and thermal properties were analysed by using X-ray diffraction and differential scanning calorimetry, respectively. There were increases in the amounts of monounsaturated and triunsaturated triacylglycerol (TAG) molecular groups with concurrent reductions of the proportions of desaturated TAG molecular groups during different time intervals. This changing TAG composition led to changes in crystallisation behaviour and thermal properties of the samples, reducing some enthalpy values. All samples withdrawn at different time intervals displayed both β' and β type crystal polymorphs even though engkabang fat itself was predominantly β‐type. In terms of melting, solidification and polymorphic properties, the sample withdrawn at 6 h time interval was found to display the closest similarity to lard (LD).
Pathogens from the Vibrio and Aeromonas genera often cause detrimental effects to the aquaculture sector. Previously, antibiotics was used to resolve the infections, but this caused the spread of antibiotic resistant bacteria and antibiotic resistance genes into the environment. As an approach to address this issue, probiotic bacteria were introduced to improve the hosts' microbiome, disease protection, health condition, growth efficiency, feed consumption, stress response and general vigour. However, reports showed that some commercially available probiotics were restricted to a small number of microbial species and there are inconsistencies concerning its effectiveness. Hence, the aim of this study was to isolate and evaluate new Bacillus spp. from the gut of giant freshwater prawn as potential probiotics. Three Bacillus spp. isolates, Bacillus subtilis FS6 (MZ960135), Bacillus velezensis FS26 (MZ960133) and Bacillus pumilus FS97 (MZ960136) were characterised, and in vitro testing showed good probiotic properties which can help in dealing with diseases in aquaculture. Among the Bacillus spp., Bacillus velezensis FS26 showed higher antimicrobial activity towards Aeromonas hydrophila LMG 13658 and Aeromonas veronii clone DK-A. veronii-27 at 23.7 mm and 25 mm, respectively. Bacillus subtilis FS6 and Bacillus velezensis FS26 resulted in good adherence to both xylene and chloroform hydrocarbons. The Bacillus spp. isolated displayed high survivability towards 0.3% bile salt and exhibited amylase, protease, and lipase activities. Thus, the isolated Bacillus spp. are considered safe based on the sensitivity analysis towards antibiotics and γ-haemolytic activity.
Bacillus velezensis FS26 is a bacterium from the genus Bacillus that has been proven as a potential probiotic in aquaculture with a good antagonistic effect on Aeromonas spp. and Vibrio spp. Whole-genome sequencing (WGS) allows a comprehensive and in-depth analysis at the molecular level, and it is becoming an increasingly significant technique in aquaculture research. Although numerous probiotic genomes have been sequenced and investigated recently, there are minimal data on in silico analysis of B. velezensis as a probiotic bacterium isolated from aquaculture sources. Thus, this study aims to analyse the general genome characteristics and probiotic markers from the B. velezensis FS26 genome with secondary metabolites predicted against aquaculture pathogens. The B. velezensis FS26 genome (GenBank Accession: JAOPEO000000000) assembly proved to be of high quality, with eight contigs containing 3,926,371 bp and an average G + C content of 46.5%. According to antiSMASH analysis, five clusters of secondary metabolites from the B. velezensis FS26 genome showed 100% similarity. These clusters include Cluster 2 (bacilysin), Cluster 6 (bacillibactin), Cluster 7 (fengycin), Cluster 8 (bacillaene), and Cluster 9 (macrolactin H), which signify promising antibacterial, antifungal, and anticyanobacterial agents against pathogens in aquaculture. The probiotic markers of B. velezensis FS26 genome for adhesion capability in the hosts' intestine, as well as the acid and bile salt-tolerant genes, were also detected through the Prokaryotic Genome Annotation System (Prokka) annotation pipeline. These results are in agreement with our previous in vitro data, suggesting that the in silico investigation facilitates establishing B. velezensis FS26 as a beneficial probiotic for use in aquaculture.
The growth performance of red claw crayfish, Cherax quadricarinatus, fed diets with different sources and concentrations of natural calcium was evaluated. Formulated diets containing 30% protein and 8% lipid were prepared using supplementation of 0 (control), 3%, 6%, 9% eggshell, and 6% seashell. C. quadricarinatus juveniles with an average total weight of 0.21 ± 0.01 g and length of 25.0 ± 0.10 mm were obtained from ten ovigerous females bred in captivity. An aquarium with a size of 0.72 m² was divided into eight compartments with an area of 0.09 m² per compartment and was individually stocked with juvenile crayfish. A total of eight juvenile crayfish were used per dietary treatment. There were five dietary treatments in total and each of these treatments were replicated three times. The addition of eggshell and seashell powder exerted positive effects on the growth performance, molting frequency, and survival of the crayfish. Crayfish fed with a 6% eggshell-supplemented diet exhibited the best overall growth performance. The calcium percentage of the carapace increased with the increase in calcium content of the diets. Meanwhile, the performance of diets comprising eggshells and seashells was not significantly different (P > 0.05). Considering the low cost and abundance of eggshells, the addition of 6% of this product to the diet is recommended for the best growth of juvenile C. quadricarinatus.
The fermentation of Malaysian fish sauce (budu) varies from one to twelve months depending on the producer, resulting in inconsistent quality. The microbiota, their predicted metabolic pathways and volatile metabolites profiles were determined at different stages of budu fermentation. Budu fermented for 1 and 3 months were characterized by the presence of Gram negative Enterobacterales, Gammaproteobacteria, and Fusobacteriaceae, which continuously decrease in abundance over fermentation time. The metabolic pathways prediction grouped 1- and 3- month budu in a cluster enriched with degradation reactions. 6-month budu were dominated by Halanaerobium and Staphylococcus, while the 12-month were dominated by Lentibacillus, Bacilli, and Halomonas. Biosynthesis-type predicted pathways involving protein and lipid derivatives were enriched in 6- and 12-month fermented budu, accumulating 2,6-dimethylpyrazine, methyl 2-ethyldecanoate, 2-phenylacetaldehyde, 3-methylbutanal, and 3-methylbutanoic acid. These compounds may indicate budu maturity and quality. This result may assist as a reference for quality control and fermentation monitoring.
This study was conducted to evaluate public awareness about COVID with aimed to check public strategies against COVID-19. A semi structured questionnaire was collected and the data was analyzed using some statistical tools (PLS-SEM) and artificial neural networks (ANN). We started by looking at the known causal linkages between the different variables to see if they matched up with the hypotheses that had been proposed. Next, for this reason, we ran a 5,000-sample bootstrapping test to assess how strongly our findings corroborated the null hypothesis. PLS-SEM direct path analysis revealed HRP -> PA-COVID, HI -> PA-COVID, MU -> PA-COVID, PM -> PA-COVID, SD -> PA-COVID. These findings provide credence to the acceptance of hypotheses H1, H3, and H5, but reject hypothesis H2. We have also examined control factors such as respondents' age, gender, and level of education. Age was found to have a positive correlation with PA-COVID, while mean gender and education level were found to not correlate at all with PA-COVID. However, age can be a useful control variable, as a more seasoned individual is likely to have a better understanding of COVID and its effects on independent variables. Study results revealed a small moderation effect in the relationships between understudy independent and dependent variables. Education significantly moderates the relationship of PA-COVID associated with MU, PH, SD, RP, PM, PA-COVID, depicts the moderation role of education on the relationship between MU*Education->PA-COVID, HI*Education->PA.COVID, SD*Education->PA.COVID, HRP*Education->PA.COVID, PM*Education -> PA.COVID. The artificial neural network (ANN) model we've developed for spreading information about COVID-19 (PA-COVID) follows in the footsteps of previous studies. The root means the square of the errors (RMSE). Validity measures how well a model can predict a certain result. With RMSE values of 0.424 for training and 0.394 for testing, we observed that our ANN model for public awareness of COVID-19 (PA-COVID) had a strong predictive ability. Based on the sensitivity analysis results, we determined that PA. COVID had the highest relative normalized relevance for our sample (100%). These factors were then followed by MU (54.6%), HI (11.1%), SD (100.0%), HRP (28.5%), and PM (64.6%) were likewise shown to be the least important factors for consumers in developing countries struggling with diseases caused by contaminated water. In addition, a specific approach was used to construct a goodness-of-fit coefficient to evaluate the performance of the ANN models. The study will aid in the implementation of effective monitoring and public policies to promote the health of local people.
ABSTRACT: Umai is a popular, traditional, native dish of the Melanau ethnic group in Sarawak. It is prepared using thin slices of raw marine fish marinated with calamansi juice and seasoned with other ingredients. The local people believe that the acidity of the citrus juice, along with the use of salt and spice, can slightly cook the fish and remove the fishy smell. The aim of this study was to investigate (i) the different umai handling and preparation practices and (ii) the personal experience of umai consumption among respondents. A purposive sample of 100 umai makers, divided into two equal groups, professionals and nonprofessionals, participated in the study. We found that Spanish mackerel and hairfin anchovy were ranked first and second in the list of species chosen for making umai, with the former mostly preferred by the professional group, as opposed to the latter, which was preferred by the nonprofessional group. Black pomfret was ranked third, where it is equally preferred by both groups. About 20% of respondents would freeze the raw fish chunks prior to preparing umai, as opposed to 26% who would sun dry their fish. Other techniques, such as salting and marinating (using calamansi juice), were also used during the preparation of umai. Most of the respondents indicated that they would consider the umai ready to eat soon after marinating (with all ingredients) the raw fish. One-third of both respondent groups indicated that they would chill the umai dish at 4°C for 30 min before serving. The respondents could not provide any rationale behind these food preparation practices. Overall, this study provides evidence of the different preparation methods for umai. These practices can thus be considered important targets for public health education campaigns seeking to improve food safety surrounding this food group.
This study aimed to isolate and identify Lactobacillus in the honey stomach of honeybee Apis dorsata. Samples of honeybee were collected from A. dorsata colonies in different bee trees and Lactobacillus bacteria isolated from honey stomachs. Ninety two isolates were Gram-stained and tested for catalase reaction. By using bacterial universal primers, the 16S rDNA gene from DNA of bacterial colonies amplified with polymerase chain reaction (PCR). Forty-nine bacterial 16S rDNA gene were sequenced and entrusted in GenBank. Phylogenetic analysis showed they were different phylotypes of Lactobacillus. Two of them were most closely relevant to the previously described species Lactobacillus plantarum. Other two phylotypes were identified to be closely related to Lactobacillus pentosus. However, only one phylotype was found to be distantly linked to the Lactobacillus fermentum. The outcomes of the present study indicated that L. plantarum, L. pentosus, and L. fermentum were the dominant lactobacilli in the honey stomach of honeybee A. dorsata collected during the dry season from Malaysia forest area - specifically "Melaleuca in Terengganu".
Isothermal amplification is a technique that can amplify target DNA sequences at a single incubation temperature. Loop-mediated isothermal amplification (LAMP) is an extension of the isothermal DNA amplification technique that combines rapidity, simplicity, high specificity and sensitivity. Due to its overwhelming characteristics, LAMP has been explored for its feasibility in detecting various subjects, and recently in meat-based food products for DNA-based meat species authentication. It has been developed to target various meat species such as porcine, chicken, horse, and ostrich with sensitivity as low as 0.1 pg/μL. Further improvement with the use of magnetic beads, electrochemiluminescence and special dye such as calcein and crystal violet had increased the sensitivity of the LAMP assay. Other important characteristics were specific target gene primers as well as a shorter incubation time, warranting a good prospect for rapid testing authentication.