Antibacterial mechanism of Malaysian Carey clay against food-borne Staphylococcus aureus.

Depending on their chemical structure and geochemistry, clay minerals can display potent antibacterial properties against a range of bacterial pathogens. Malaysian Carey clay was evaluated for its antibacterial activity against food-borne Staphylococcus aureus ATCC 13565 strains. The minimum inhibitory concentration (MIC) and minimum bactericidal activity (MBC) of both Carey clay leachates and suspension were 125 mg/mL and 250 mg/mL, respectively. Time-kill assay revealed that 2x MIC and 4x MIC Carey clay in both leachate and suspension forms resulted in complete killing of S. aureus. Antibacterial mechanism was investigated through imaging of bacterial morphology using TEM and determination of reactive oxygen species (ROS) using NBT assay. Imaging of bacterial morphology using TEM showed abnormalities, including disrupted cell walls following exposure to Carey clay, and the antibacterial activity was associated with generation of ROS. Our study suggests that Carey clay displays promising functionality as a natural antibacterial agent in the food industry.

A review of CRISPR-Cas and PCR-based methods for the detection of animal species in the food chain-current challenges and future prospects.

Regular testing and systematic investigation play a vital role to ensure product safety. Until now, the existing food authentication techniques have been based on proteins, lipids, and nucleic acid-based assays. Among various deoxyribonucleic acid (DNA)-based methods, the recently developed Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) based bio-sensing is an innovative and fast-expanding technology. The CRISPR/Cas-9 is known as Clustered Regularly Interspaced Short Palindromic Repeats due to the flexibility and simplicity of the CRISPR/Cas9 site-specific editing tool has been applied in many biological research areas such as Gene therapy, cell line development, discovering mechanisms of disease, and drug discovery. Nowadays, the CRISPR-Cas system has also been introduced into food authentication via detecting DNA barcodes of poultry and livestock both in processed and unprocessed food samples. This review documents various DNA based approaches, in an accessible format. Future CRISPR technologies are forecast while challenges are outlined.

Effect of natural antibacterial clays against single biofilm formation by Staphylococcus aureus and Salmonella Typhimurium bacteria on a stainless-steel surface.

Staphylococcus aureus and Salmonella Typhimurium have a propensity to develop biofilms on food contact surfaces, such as stainless-steel, that persist despite rigorous cleaning and sanitizing procedures. Since both bacterial species pose a significant public health risk within the food chain, improved anti-biofilm measures are needed. This study examined the potential of clays as antibacterial and anti-biofilm agents against these two pathogens on appropriate contact surfaces. Natural soil was processed to yield leachates and suspensions of both untreated and treated clays. Soil particle size, pH, cation-exchange capacity, and metal ions were characterized to assess their importance in bacterial killing. Initial antibacterial screening was performed on nine distinct types of natural Malaysian soil using a disk diffusion assay. Untreated leachate from Kuala Gula and Kuala Kangsar clays were found to inhibit S. aureus (7.75 ± 0.25 mm) and Salmonella Typhimurium (11.85 ± 1.63 mm), respectively. The treated Kuala Gula suspension (50.0 and 25.0 %) reduced S. aureus biofilms by 4.4 and 4.2 log at 24 and 6 h, respectively, while treated Kuala Kangsar suspension (12.5 %) by a 4.16 log reduction at 6 h. Although less effective, the treated Kuala Gula leachate (50.0 %) was effective in removing Salmonella Typhimurium biofilm with a decrease of >3 log in 24 h. In contrast to Kuala Kangsar clays, the treated Kuala Gula clays contained a much higher soluble metal content, especially Al (301.05 ± 0.45 ppm), Fe (691.83 ± 4.80 ppm) and Mg (88.44 ± 0.47 ppm). Elimination of S. aureus biofilms correlated with the presence of Fe, Cu, Pb, Ni, Mn and Zn irrespective of the pH of the leachate. Our findings demonstrate that a treated suspension is the most effective for eradication of S. aureus biofilms with a potential as a sanitizer-tolerant, natural antibacterial against biofilms for applications in the food industry.

Microbiological and Sensorial Quality of Beef Meat (Longissimus dorsi) Marinated with Cinnamon Extract and Stored at Various Temperatures.

Meat spoilage caused by temperature abuse is a major problem for producers, retailers, and consumers that can generate large economic losses to industries. Microbial growth of Pseudomonas spp. is the main source of spoilage during storage. Cinnamon has antimicrobial properties that may potentially be used to reduce the spoilage caused by Pseudomonas. The objectives of this study were to determine the inhibitory effect of cinnamon extract (CE) against Pseudomonas aeruginosa (ATCC 27853) and evaluate the treatment of CE on meat quality during different storage temperatures (5 °C, 10 °C, 15 °C, and 25 °C). The anti-Pseudomonas result showed that 100% (w/v) CE concentration produced a 13.50 mm zone of inhibition in a disc diffusion assay. The minimum inhibitor concentration (MIC) of CE was noted at 25% (v/v), whereas the minimum bactericidal concentration (MBC) value was observed at 50% (v/v) concentration of CE. The time-kill showed the growth of P. aeruginosa decreased from 7.64 to 5.39 log CFU/mL at MIC concentration. Total phenolic content and IC50 value of the cinnamon extract was expressed as 6.72 ± 0.87 mg GAE/g extract and 0.15 mg/mL, respectively. When the meat was marinated with 50% (v/v) CE and stored at various temperatures, the total viable count (TVC) and growth of Pseudomonas spp. were lowered as compared to the control sample. However, the reduction in microbial count in all samples was influenced by the storage temperature, where the lowered microbial count was noted in the sample treated with CE and stored at 5 and 10 °C for 48 h. The pH of meat treated with or without CE ranged from pH 5.74 to 6.48. The sensory attributes of colour, texture, and overall acceptability have a significant difference, except for odour, between marinated meat and control. The results indicate that the use of cinnamon extract as the marination agent for meat could reduce the growth of Pseudomonas spp. and therefore assist in extending the shelf life of meat at 5 and 10 °C storage temperatures.

Aflatoxin Biosynthesis, Genetic Regulation, Toxicity, and Control Strategies: A Review.

Aflatoxins (AFs) are highly toxic and cancer-causing compounds, predominantly synthesized by the Aspergillus species. AFs biosynthesis is a lengthy process that requires as minimum as 30 genes grouped inside 75 kilobytes (kB) of gene clusters, which are regulated by specific transcription factors, including aflR, aflS, and some general transcription factors. This paper summarizes the status of research on characterizing structural and regulatory genes associated with AF production and their roles in aflatoxigenic fungi, particularly Aspergillus flavus and A. parasiticus, and enhances the current understanding of AFs that adversely affect humans and animals with a great emphasis on toxicity and preventive methods.

Biocontrol of Aflatoxins Using Non-Aflatoxigenic Aspergillus flavus: A Literature Review.

Aflatoxins (AFs) are mycotoxins, predominantly produced by Aspergillus flavus, A. parasiticus, A. nomius, and A. pseudotamarii. AFs are carcinogenic compounds causing liver cancer in humans and animals. Physical and biological factors significantly affect AF production during the pre-and post-harvest time. Several methodologies have been developed to control AF contamination, yet; they are usually expensive and unfriendly to the environment. Consequently, interest in using biocontrol agents has increased, as they are convenient, advanced, and friendly to the environment. Using non-aflatoxigenic strains of A. flavus (AF-) as biocontrol agents is the most promising method to control AFs' contamination in cereal crops. AF- strains cannot produce AFs due to the absence of polyketide synthase genes or genetic mutation. AF- strains competitively exclude the AF+ strains in the field, giving an extra advantage to the stored grains. Several microbiological, molecular, and field-based approaches have been used to select a suitable biocontrol agent. The effectiveness of biocontrol agents in controlling AF contamination could reach up to 99.3%. Optimal inoculum rate and a perfect time of application are critical factors influencing the efficacy of biocontrol agents.

Antibacterial Activity of Clay Soils against Food-Borne Salmonella typhimurium and Staphylococcus aureus.

Natural clays have recently been proven to possess antibacterial properties. Effective natural antimicrobial agents are needed to combat bacterial contamination on food contact surfaces, which are increasingly more prevalent in the food chain. This study sought to determine the antibacterial activity of clays against the food-borne pathogens Salmonella typhimurium ATCC 14028 and Staphylococcus aureus ATCC 13565. Soils were processed to yield leachates and suspensions from untreated and treated clays. Soil particle size, pH, cation-exchange capacity, metal composition and mineralogy were characterized. Antibacterial screening was performed on six Malaysian soils via the disc diffusion method. In addition, a time-kill assay was conducted on selected antibacterial clays after 6 h of exposure. The screening revealed that Munchong and Carey clays significantly inhibit Salmonella typhimurium (11.00 ± 0.71 mm) and S. aureus (7.63 ± 0.48 mm), respectively. Treated Carey clay leachate and suspension completely kill Salmonella typhimurium, while S. aureus viability is reduced (2 to 3 log10). The untreated Carey and all Munchong clays proved ineffective as antibacterials. XRD analysis confirmed the presence of pyrite and magnetite. Treated Carey clays had a higher soluble metal content compared to Munchong; namely Al (92.63 ± 2.18 mg/L), Fe (65.69 ± 3.09 mg/L) and Mg (88.48 ± 2.29 mg/L). Our results suggest that metal ion toxicity is responsible for the antibacterial activity of these clays.

Optimization of γ-aminobutyric acid production by Lactobacillus plantarum Taj-Apis362 from honeybees.

Dominant strains of lactic acid bacteria (LAB) isolated from honey bees were evaluated for their γ-aminobutyric acid (GABA)-producing ability. Out of 24 strains, strain Taj-Apis362 showed the highest GABA-producing ability (1.76 mM) in MRS broth containing 50 mM initial glutamic acid cultured for 60 h. Effects of fermentation parameters, including initial glutamic acid level, culture temperature, initial pH and incubation time on GABA production were investigated via a single parameter optimization strategy. The optimal fermentation condition for GABA production was modeled using response surface methodology (RSM). The results showed that the culture temperature was the most significant factor for GABA production. The optimum conditions for maximum GABA production by Lactobacillus plantarum Taj-Apis362 were an initial glutamic acid concentration of 497.97 mM, culture temperature of 36 °C, initial pH of 5.31 and incubation time of 60 h, which produced 7.15 mM of GABA. The value is comparable with the predicted value of 7.21 mM.

Effect of supercritical fluid extraction on the reduction of toxic elements in fish oil compared with other extraction methods.

High-quality fish oil for human consumption requires low levels of toxic elements. The aim of this study was to compare different oil extraction methods to identify the most efficient method for extracting fish oil of high quality with the least contamination. The methods used in this study were Soxhlet extraction, enzymatic extraction, wet reduction, and supercritical fluid extraction. The results showed that toxic elements in fish oil could be reduced using supercritical CO2 at a modest temperature (60°C) and pressure (35 MPa) with little reduction in the oil yield. There were significant reductions in mercury (85 to 100%), cadmium (97 to 100%), and lead (100%) content of the fish oil extracted using the supercritical fluid extraction method. The fish oil extracted using conventional methods contained toxic elements at levels much higher than the accepted limits of 0.1 µg/g.

Linearity study on detection and quantification limits for the determination of avermectins using linear regression.

In this study, linear relationships between response and concentration were used to estimate the detection limit (DL) and quantification limit (QL) for five avermectins: emamectin, abamectin, doramectin, moxidectin, and ivermectin. Estimation of DL and QL was based on the standard deviation of residual and y-intercept of the regression line at low concentrations of avermectins, using the dispersive solid-phase extraction procedure. Avermectin extracts were analyzed using liquid chromatography tandem mass spectrometry. Based on the regression slope, DL and QL were higher at concentrations of 0.3-0.4 µg/kg and 1 µg/kg, respectively, for all avermectin compounds. Linearity assessment was performed by linear regression, which incorporated a regression model, outlier rejection, and evaluation of the assumption with a significant test. For all avermectins, there is a significant correlation between response and concentration in the range 1-15 µg/kg, and the y-intercept passes through origin (zero).

Prevalence and quantification of Listeria monocytogenes in beef offal at retail level in Selangor, Malaysia

A total of 63 beef offal samples (beef liver = 16; beef lung = 14; beef intestine = 9; beef tripe = 15; beef spleen = 9) from three wet markets (A, B, and C) in Selangor, Malaysia were examined for the prevalence and microbial load of Listeria monocytogenes. A combination of the most probable number and polymerase chain reaction (MPN-PCR) method was employed in this study. It was found that L. monocytogenes detected in 33.33% of the beef offal samples. The prevalence of L. monocytogenes in beef offal purchased from wet markets A, B, and C were 22.73%, 37.50% and 41.18% respectively. The density of L. monocytogenes in all the samples ranged from<3upto> 2,400 MPN/g. The findings in this study indicate that beef offal can be a potential vehicle of foodborne listeriosis.

Prevalence and quantification of Listeria monocytogenes in beef offal at retail level in Selangor, Malaysia.

A total of 63 beef offal samples (beef liver = 16; beef lung = 14; beef intestine = 9; beef tripe = 15; beef spleen = 9) from three wet markets (A, B, and C) in Selangor, Malaysia were examined for the prevalence and microbial load of Listeria monocytogenes. A combination of the most probable number and polymerase chain reaction (MPN-PCR) method was employed in this study. It was found that L. monocytogenes detected in 33.33% of the beef offal samples. The prevalence of L. monocytogenes in beef offal purchased from wet markets A, B, and C were 22.73%, 37.50% and 41.18% respectively. The density of L. monocytogenes in all the samples ranged from < 3 up to > 2,400 MPN/g. The findings in this study indicate that beef offal can be a potential vehicle of foodborne listeriosis.

Mercury pollution in Malaysia.

Although several studies have been published on levels of mercury contamination of the environment, and of food and human tissues in Peninsular Malaysia, there is a serious dearth of research that has been performed in East Malaysia (Sabah and Sarawak). Industry is rapidly developing in East Malaysia, and, hence, there is a need for establishing baseline levels of mercury contamination in environmental media in that part of the country by performing monitoring studies. Residues of total mercury and inorganic in food samples have been determined in nearly all previous studies that have been conducted; however, few researchers have analyzed samples for the presence of methlymercury residues. Because methylmercury is the most toxic form of mercury, and because there is a growing public awareness of the risk posed by methylmercury exposure that is associated with fish and seafood consumption, further monitoring studies on methylmercury in food are also essential. From the results of previous studies, it is obvious that the economic development in Malaysia, in recent years, has affected the aquatic environment of the country. Primary areas of environmental concern are centered on the rivers of the west Peninsular Malaysian coast, and the coastal waters of the Straits of Malacca, wherein industrial activities are rapidly expanding. The sources of existing mercury input to both of these areas of Malaysia should be studied and identified. Considering the high levels of mercury that now exists in human tissues, efforts should be continued, and accelerated in the future, if possible, to monitor mercury contamination levels in the coastal states, and particularly along the west Peninsular Malaysian coast. Most studies that have been carried out on mercury residues in environmental samples are dated, having been conducted 20-30 years ago; therefore, the need to collect much more and more current data is urgent. Furthermore, establishing baseline levels of mercury exposure to humans in Malaysia will be useful in establishing the levels at which detrimental effects in both humans and marine life may occur, and therefore the levels at which warning should be raised or limits established. In particular, we believe that two or three monitoring centers should be established in Peninsular Malaysia, and one in East Malaysia for the specific purpose of monitoring for the presence of hazardous environmental chemicals, and particularly monitoring for heavy metals such as mercury that reach food that is subject to consistent human consumption.

The isolation of a new S-methyl benzothioate compound from a marine-derived Streptomyces sp.

The application of an HPLC bioactivity profiling/microtiter plate technique in conjunction with microprobe NMR instrumentation and access to the AntiMarin database has led to the isolation of a new 1. In this example, 1 was isolated from a cytotoxic fraction of an extract obtained from marine-derived Streptomyces sp. cultured on Starch Casein Agar (SCA) medium. The 1D and 2D (1)H NMR and ESIMS data obtained from 20 µg of compound 1 fully defined the structure. The known 2 was also isolated and readily dereplicated using this approach.