Evaluation of the microplastics in bivalves and water column at Pantai Teluk Likas, North Borneo, Malaysia.

Microplastics (MPs) are a pervasive pollutant in the marine environment. Pantai Teluk Likas in Sabah, Malaysia is one of the most visited beaches where tourism, recreational, and fisheries activities are high in this area. Hence, the area suffers from severe pollution, particularly from plastics. This study aims to quantify the microplastic composition in terms of color, shapes, and polymer types in marine bivalves (Anadara granosa, Glauconome virens, and Meretrix lyrata) and water column samples from Pantai Teluk Likas. All samples were digested using sodium hydroxide (NaOH) and incubated in the oven for at least 48 h. Serial filtration was done for each sample before they were observed under the dissecting microscope. The microplastics were identified and counted based on their physical attributes which were colors and shapes. The functional group of the polymers was determined using FTIR spectroscopy. Microplastics were found present in all samples collected. G. virens had the highest abundance of microplastics at 113.6 ± 6.5 particles/g followed by M. lyrata at 78.4 ± 3.7 particles/g. On the contrary, A. granosa had the least microplastics with an abundance of 24.4 ± 0.6 particles/g. Meanwhile, 110.0 ± 36.2 particles/L of microplastics were found in water column samples from Pantai Teluk Likas. Based on the analysis, fibers were the most common shape in bivalves, while fibers and films were common in the water column. In terms of colors, black, blue, and red were a few of the most abundant colors observed in both samples. The most common polymer detected in all bivalve species and water column samples is polycarbonate (PC), followed by polymethyl methacrylate (PMMA). Future study that focuses on the correlation between microplastic abundance in the marine biota and the water column is recommended to better understand microplastic availability and exposure.

Evaluation of microplastics isolated from sea cucumber Acaudina molpadioides in Pulau Langkawi, Malaysia.

Plastic pollution is an emerging environmental concern in recent years due to continuous mass production and its slow degradation. Microplastics measuring between 5 mm and 1 µm are being ingested by marine animals and eventually by human consumption in form of seafood. The aim of this research was to evaluate microplastics isolated from sea cucumber Acaudina molpadioides in Pulau Langkawi. A total of 20 animals were collected and their gastrointestinal tract were digested using NaOH. Microplastics were isolated, filtered and identified through microscopic examination based on the colour, shape and size. The chemical composition of microplastics were further analyzed by FTIR to identify the functional group of polymers. A total of 1652 microplastics were found in A. molpadioides. Fibres (99.4%) and black color (54.4%) were the majority of microplastics observed in terms of shapes and colors. The size range within 0.5-1 µm and 1-2 µm were the highest abundance observed. There were two identified polymer types of microplastics obtained through FTIR which were polyethylene (PE) and polymethyl methacrylate (PMMA). In conclusion, microplastics were isolated from the gastrointestinal tract of A. molpadioides indicating that the animals were contaminated. Further research can be done on the toxicity effects of these microplastics towards human upon consumption of these animals as seafood.

Metabolites and Bioactivity of the Marine Xestospongia Sponges (Porifera, Demospongiae, Haplosclerida) of Southeast Asian Waters.

Sponges are aquatic, spineless organisms that belong to the phylum Porifera. They come in three primary classes: Hexactinellidae, Demospongiae, and Calcarea. The Demospongiae class is the most dominant, making up over 90% of sponge species. One of the most widely studied genera within the Demospongiae class is Xestospongia, which is found across Southeast Asian waters. This genus is of particular interest due to the production of numerous primary and secondary metabolites with a wide range of biological potentials. In the current review, the antioxidant, anticancer, anti-inflammatory, antibacterial, antiviral, antiparasitic, and cytotoxic properties of metabolites from several varieties of Southeast Asian Xestospongia spp. were discussed. A total of 40 metabolites of various natures, including alkaloids, fatty acids, steroids, and quinones, were highlighted in X. bergquistia, X. testudinaria, X. muta, X. exigua, X. ashmorica and X. vansoesti. The review aimed to display the bioactivity of Xestospongia metabolites and their potential for use in the pharmaceutical sector. Further research is needed to fully understand their bioactivities.

In Vitro Assessment on Designing Novel Antibiofilms of Pseudomonas aeruginosa Using a Computational Approach.

An anti-biofilm that can inhibit the matrix of biofilm formation is necessary to prevent recurrent and chronic Pseudomonas aeruginosa infection. This study aimed to design compounds with a new mechanism through competitive inhibitory activity against phosphomannomutase/phosphoglucomutase (PMM/PGM), using in vitro assessment and a computational (in silico) approach. The active site of PMM/PGM was assessed through molecular redocking using L-tartaric acid as the native ligand and other small molecules, such as glucaric acid, D-sorbitol, and ascorbic acid. The docking program set the small molecules to the active site, showing a stable complex formation. Analysis of structural similarity, bioavailability, absorption, distribution, metabolism, excretion, and toxicity properties proved the potential application of ligands as an anti-biofilm. In vitro assessment with crystal violet showed that the ligands could reach up to 95.87% inhibition at different concentrations. The nitrocellulose membrane and scanning electron microscopic visualization showed that the untreated P. aeruginosa biofilm was denser than the ligand-treated biofilm.

An improved Fourier-Transform Infrared Spectroscopy combined with partial least squares regression for rapid quantification of total aflatoxins in commercial chicken feeds and food grains.

Objective: This study aims to develop and validate an Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy (ATR-FTIR) spectroscopic technique combined with a partial least squares regression (PLSR) model for rapid quantification and monitoring of aflatoxins in chicken feeds and food grains. Materials and Methods: A model of ATR-FTIR-PLSR was developed using ATR-FTIR spectra of mixed aflatoxin standards in 100% acetonitrile (112 samples) and 75% methanol (112 samples), validated by testing its prediction on 125 feed/food samples spiked with variable concentrations of aflatoxins, and applied to screen 660 samples of commercial chicken feeds and food grains from Nigerian and Malaysian markets for total aflatoxins, for which the dietary exposure risks to aflatoxins (DERA) and associated hepatocellular carcinoma (HCC) risks were evaluated for both countries. Results: The ATR-FTIR-PLSR model demonstrated excellent prediction power [R 2 = 99.59%, p = 0.001, root mean square error of calibration (RMSEC) = 1.69, RMSE p = 1.98, bias = -0.26], sensitivity (limit of quantitation and limit of the method < 5.0 ng/gm), precision (coefficient of variation = 0.97-1.72), and accuracy (% recovery of 88%-106%) in all the spiked samples. The model's prediction was statistically reliable (R 2 = 99.8%, p < 0.05) when compared with a high-performance liquid chromatography method. Levels of aflatoxins in the commercial samples signify high DERA (0.92-138.2 ng of aflatoxins/kg BW/day) and HCC risk (1.07%-159.91% of HCC/100,000 people/year) in the exposed populations. Conclusions: Results feature the conceivable implementation of the proposed ATR-FTIR-PLSR model for rapid, accurate determination and monitoring of aflatoxins in commercial chicken feeds and food grains; and the need to strengthen aflatoxin control/prevention strategies in the study populations.

Ultra-fast RP-HPLC-FD-DAD for quantification of total aflatoxins in maize, rice, wheat, peanut and poultry feed without sample clean up, and population exposure risk assessment in Katsina, Nigeria: an optimization study.

This study reports the development and validation of a simple, yet efficient method called the ultra-fast reverse phase high-performance liquid chromatography with fluorescence and photodiode array detector (UF-RP-HPLC-FD-DAD) to extract and quantify the total aflatoxin from grains and poultry feed. The proposed method is used to determine the total aflatoxin content in 150 samples of maize, rice, wheat, peanut and poultry feed obtained from open markets in a state in Nigeria. The extent of consumer exposure to aflatoxins and the risk of developing hepatocellular carcinoma (HCC) are evaluated. The UF-RP-HPLC-FD-DAD method was found to be satisfactorily accurate, sensitive and reliable as ascertained by its excellent validation outcomes (R2 > 0.999, LoD < 0.08 ng g-1, LoQ < 0.2 ng g-1, recovery = 90-102%). The aflatoxin levels in food grains and poultry feed samples obtained in this study implied a moderate dietary exposure of between 10.67 and 20.77 ng/kg BW/day, in which the risk of developing HCC was estimated to be between 6.27 and 21.40% per 100,000 adults/year. Hence, greater monitoring of marketed food and feed is required, besides the deployment of strict controls and preventive techniques to minimize the population's exposure to a high dietary level of aflatoxins.

Evaluation of microplastics ingested by sea cucumber Stichopus horrens in Pulau Pangkor, Perak, Malaysia.

The widespread presence of microplastics has caused significant concerns on their potential effects on marine ecosystems. Microplastics are classified as plastic products of less than 5 mm in size and are known to be one of the most dangerous aquatic debris to marine species. Sea cucumbers are deposit feeders living in sediment regions and benthic zone that may collect microplastics in low-energy environments. This research aims to evaluate the types of microplastic isolated from the intestine of Stichopus horrens in Malaysia. This species is selected because it is mainly found in this island. Pulau Pangkor is one of the leading tourism destinations on the small islands is facing increasing numbers of tourists and development and it is useful for sustainable study. A total of 20 S. horrens samples collected from Pulau Pangkor, Malaysia were dissected, and their intestines were collected for digestion. Microplastics were extracted using NaOH and filtered using filter paper. Microplastic identification was conducted on the based of the physical characteristics (colour, shape, size) and chemical characterisation was evaluated using FTIR for polymer functional groups. A total of 1446 unit of microplastics were found in S. horrens. Among various types and colours, fibre (90%) and black (59%) were dominant amongst the various particles identified. The majority of microplastics sizes were 0.51 µm and 1-2 µm. Two polymer materials were identified, namely polyethylene and poly(methyl methacrylate). As a conclusion, the findings of the study will serve as primary data for pollution indicators in respective islands. The microplastics could impact the immune system and upset gut's balance of human.

Development and validation of quantitative thin layer chromatographic technique for determination of total aflatoxins in poultry feed and food grains without sample clean-up.

OBJECTIVE: The purpose of this work is to develop and validate an appropriate solvent solution and quantitative thin layer chromatography (TLC) method for determining the aflatoxins content of chicken feeds and dietary grains. MATERIALS AND METHODS: To obtain the optimal mobile phase, samples were extracted with methanol/water (3:1) + 5% sodium chloride and partitioned using several solvent systems using preparative TLC. Camag TLC scanner 3 was used to scan the TLC plates at 366 nm and quantify them using JustTLC software. The method was tested for linearity, specificity, accuracy, precision, sensitivity, and robustness in accordance with ICH recommendations, and then utilized to screen 132 Nigerian poultry/food samples for total aflatoxins (TAFs). RESULTS: The best separation of aflatoxins was achieved using acetonitrile and dichloromethane (3:17) mobile phase over an average run time of 45 min, resulting in linear calibration curves (R2 > 0.99) in the concentration range limit of quantitation (LoQ) to 50 ng/spot with a limit of detection of <2.0 ng/g and a LoQ of <4.0 ng/gm for all aflatoxins in all spiked samples. When the proposed TLC method was compared to an optimized high-performance liquid chromatography method, an excellent linear regression was obtained (R2 > 95%). Seventy seven (58.33%) of the 132 samples examined were positive for aflatoxins, with mean values ranging from 3.57 ± 2.55 to 37.31 ± 34.06 ng/gm for aflatoxin B1 and 6.67 ± 0.00 to 38.02 ± 31.52 ng/gm for TAFs, respectively. CONCLUSIONS: The results demonstrate the feasibility of using the suggested TLC method in conjunction with a novel solvent solution (free of carcinogenic chloroform) for the rapid and accurate measurement of TAFs in foods/feeds.

Prevalence, Virulence Genes and Antimicrobial Resistance Profiles of Salmonella Serovars from Retail Beef in Selangor, Malaysia.

The aim of the present study was to investigate the prevalence of Salmonella spp., Salmonella Enteritidis and Salmonella Typhimurium in retail beef from different retail markets of Selangor area, as well as, to assess their pathogenic potential and antimicrobial resistance. A total of 240 retail beef meat samples (chuck = 60; rib = 60; round = 60; sirloin = 60) were randomly collected. The multiplex polymerase chain reaction (mPCR) in combination with the most probable number (MPN) method was employed to detect Salmonella spp., S. Enteritidis and S. Typhimurium in the meat samples. The prevalence of Salmonella spp., S. Enteritidis and S. Typhimurium in 240 beef meat samples were 7.50, 1.25, and 0.83%, respectively. The microbial loads of total Salmonella was found in the range of <3 to 15 MPN/g. Eight different serovars of Salmonella were identified among the 23 isolates, and S. Agona was the predominant serovar (26.09%). Interestingly, all the Salmonella isolates were resistant to penicillin, erythromycin and vancomycin, but the sensitivity was observed for tetracycline, gentamicin and amoxicillin/clavulanic acid. All 23 isolates were resistant to at least three antibiotics. Two S. Typhimurium isolates (8.70%) exhibited the highest multiple antibiotic resistance (MAR) index value of 0.56 which shown resistance to nine antibiotics. PCR analysis of virulence genes showed that all Salmonella isolates (100%) were positive for the invA gene. Meanwhile, pefA was only identified in S. Enteritidis and S. Typhimurium. The findings in this study indicate that retail beef products tested were widely contaminated with multi-drug resistant (MDR) Salmonella and various virulence genes are present among the isolated Salmonella serovars.

Toxicity effects of water extracts of Holothuria atra Jaeger in mice.

OBJECTIVE: To determine lethal median dose (LD50) and histopathological toxicity of water extract of Holothuria atra (H. atra) in mice. METHODS: The behavioral changes, mortality and histopathology examination on liver were assessed in mice 14 d after the administration (i.p.) of H. atra water extract. Seven doses (10, 20, 30, 50, 100, 150 and 200 mg/kg) of H. atra were used. The control group was treated with normal saline. RESULTS: In the acute study in mice, the water extracts of H. atra caused dose-dependent general behavior adverse affects and mortality. The main behavioral sign of toxicity was hypoactivity, noticed immediately after administration of the extract which was more obvious at the higher doses and persisted until death. Mortality increased with increasing doses, the calculated LD50 was 41 mg/kg in mice. The liver toxicity was confirmed by histopathological examination, which indicated the presence of abnormal hepatocytes with a distorted shape and undefined cell lining as well as enlarged nuclei in low doses groups. High doses groups indicated a more prominent distortion of the polyhedral hepatocytes with undefined cell lining, massive cytoplasm, pyknotic, karyorhexis and karyolytic nuclei (necrosis of hepatocytes). Control group showed polyhedral hepatocytes with defined cell lining arranged in cords and normal round nuclei, with granular cytoplasm. CONCLUSIONS: Because of the relatively low LD50 value in the acute study in mice, it may be concluded that the H. atra water extract is toxic.

Toxicity effects of water extracts of Holothuria atra Jaeger in mice

Objective:To determine lethal median dose (LD50) and histopathological toxicity of water extract of Holothuria atra (H. atra) in mice. Methods: The behavioral changes, mortality and histopathology examination on liver were assessed in mice 14 d after the administration (i.p.) of H. atra water extract. Seven doses (10, 20, 30, 50, 100, 150 and 200 mg/kg) of H. atra were used. The control group was treated with normal saline. Results:In the acute study in mice, the water extracts of H. atra caused dose-dependent general behavior adverse affects and mortality. The main behavioral sign of toxicity was hypoactivity, noticed immediately after administration of the extract which was more obvious at the higher doses and persisted until death. Mortality increased with increasing doses, the calculated LD50 was 41 mg/kg in mice. The liver toxicity was confirmed by histopathological examination, which indicated the presence of abnormal hepatocytes with a distorted shape and undefined cell lining as well as enlarged nuclei in low doses groups. High doses groups indicated a more prominent distortion of the polyhedral hepatocytes with undefined cell lining, massive cytoplasm, pyknotic, karyorhexis and karyolytic nuclei (necrosis of hepatocytes). Control group showed polyhedral hepatocytes with defined cell lining arranged in cords and normal round nuclei, with granular cytoplasm. Conclusions: Because of the relatively low LD50 value in the acute study in mice, it may be concluded that the H. atra water extract is toxic.