The Salmonella outbreak is one of the leading foodborne diseases in the world withincreasing cases being reported annually. However, the current methods for Salmonelladetection in foods are outdated, laborious and time-consuming. This necessitateddeveloping a technique that is rapid for Salmonella detection in foods. Thus, the currentstudy aimed to develop a multiplex touchdown PCR (m-TdPCR) protocol for rapid andsimultaneous detection of Salmonella enterica subsp. enterica serovars Enteritidis andTyphimurium in foods. A two-phase m-TdPCR protocol was developed and optimizedwith primer pairs targeting the Salmonella enterica subsp. enterica (ST11/ST15-0.15 µM),serovars Enteritidis (sdfI gene-1.2 µM), Typhimurium (fliC gene-1.5 µM) and an internalamplification control (16S rRNA-0.08 µM). It was found that the m-TdPCR protocol ishighly sensitive detecting up to 1 ng of Salmonella DNA and its specificity was verifiedusing the in-silico method. Furthermore, the developed m-TdPCR shows no non-specificPCR amplicons and is able to detect both S. enterica ser. Enteritidis and S. enterica ser.Typhimurium in real-time when tested against the artificially contaminated food samplesat up to 10-3dilutions. Therefore, the validated m-TdPCR protocol in this study can beused as a tool for rapid detection of S. enterica ser. Enteritidis and S. enterica ser.Typhimurium in food samples and this may significantly reduce any related foodborneincidences in future.

Marine animals are continuously exposed to a wide range of microorganisms present in their environment. Studies indicated that the microorganisms could be found in different parts of the animal’s body. There are different types of lactic acid bacteria in various isolation sites and studies have disclosed the presence of Lactobacillus, Lactococcus, Enterococcus, Streptococcus, Vagococcus, Pediococcus, Weissella, Leuconostoc, and Carnobacterium. The lactic acid bacteria (LAB) of these animals have been the subject of various studies. They were studied for their inhibitory activity against pathogens, tested for their antibiotic resistance, and searched for the production of bacteriocin which some are eventually made into commercial products. This study reviewed isolated LAB from marine animals to discover the better potential of LAB that could be implemented in different industries such as food and beverages, pharmaceutical, aquaculture, nutraceutical, and medical. This review assembled the species of LAB that were isolated from different parts of the marine animals’ bodies and their application.

Presence of Salmonella in organic farming may lead to contamination in fresh produce. This study was designed to detect Salmonella contamination in organic vegetable farm situated in Serdang and to evaluate the antibiotic susceptibility profiles of the isolates. A total of 460 samples of brinjal, cucumber, ladies’ fingers and soil were collected and examined for the presence of Salmonella. The obtained isolates were identified and confirmed by biochemical characterization and serotyping. Antibiotic sensitivity profiles of the isolates were determined by using agar disk diffusion method. Salmonella spp. was detected in brinjal (1.7%, n=2), cucumber (1.7%, n=2), ladies’ fingers (2.5%, n=3) and soil (5.0%, n=5) samples. The prevalent serovars were Salmonella enterica ser. Senftenberg, S. enterica ser. Weltevreden and S. enterica ser. Corvallis. All strains were resistance to penicillin and vancomycin, with multiple antibiotic resistance (MAR) index between 0.21 and 0.36, demonstrated here as multi-drug resistant (MDR) Salmonella. The result highlighted that organic vegetables constituted potential sources of Salmonella, informing continuous monitoring and tightened surveillance are necessary to ensure food safety.

Pennywort (Centella asiatica) is a herbaceous vegetable commonly consumed raw as 'ulam' or salad. Consumption of raw leafy green vegetables is one of the pathogenic mechanisms that could cause foodborne outbreaks. The aim of the present work was therefore to investigate the effect of pulsed light (PL) treatment at fluences of 1.5, 4.2, 6.9, 9.6, and 12.3 J/cm(2) on the microbiological and physical quality of pennywort stored at 4 +/- 1 degrees C. Escherichia coli (E. coli) were inoculated onto the pennywort leaves before being exposed to PL and viewed using scanning electron microscopy (SEM). PL fluences of 6.9, 9.6, and 12.3 J/cm(2) significantly reduced the microbial count; however, the highest inactivation was obtained by using fluences of 9.6 and 12.3 J/cm(2). The color of pennywort was not significantly affected by PL treatment applied at lower fluences of 1.5, 4.2, and 6.9 J/cm(2); however, at higher fluence, 9.6 and 12.3 J/cm(2), the color was affected. PL at 1.5, 4.2, 6.9, and 9.6 J/cm(2) was able to retain the texture appearance of the leaves. To conclude, PL at 6.9 J/cm(2) showed the best fluence to reduce total aerobic mesophilic count while retaining the physical properties of pennywort leaves and extend the shelf life to about four days. The inactivation of E. coli population was significantly higher at PL fluence of 6.9 J/cm(2). It was observed that PL caused the destruction to the surface of E. coli's cell membrane. The reductions of samples inoculated with E. coli were better than those achieved in native microbiota. Furthermore, the present work also demonstrated that PL treatment was able to reduce the microbial count on pennywort leaves. (C) All Rights Reserved

Short mackerel (Rastrelliger brachysoma) is a common fish consumed in Malaysia. The high protein content of fish makes it prone to contamination with bacteria, including Listeria monocytogenes. Therefore, the objective of this study is to control the growth of L. monocytogenes by marinating short mackerel with turmeric-salt and storing at various temperatures. The short mackerel was treated with 70% of ethanol before marinating with turmeric-salt in a ratio (1:1). The short mackerel were inoculated with a concentration of 108 CFU/mL of L. monocytogenes and was stored at 0, 5, 10, 15, 20, and 25˚C for 21 days. The total microbial count and L. monocytogenes count were determined for every three days from 0 days to 21 days. The growth of L. monocytogenes in marinated mackerel was from 4.30 log CFU/g (0 days at 0 ˚C) to 5.22 log CFU/g (0 days at 25 ˚C). While the total of L. monocytogenes counts in non-marinated (control), short mackerel increased significantly based on temperature and time from 4.98 log CFU/g (0 days at 0 ˚C) to 6.22 log CFU/g (0 days at 25 ˚C). These results indicated that the growth of L. monocytogenes in the presence of turmeric-salt slowed compared with the non-marinated product, although the growth of L. monocytogenes was gradually increased throughout 21 days of storage. There was a significant difference (p<0.05) in the growth of L. monocytogenes with the storage temperature. In conclusion, the turmeric-salt marination has the potential to slow the growth of L. monocytogenes in short mackerel by approximately 0.07 log CFU/g - 2.81 log CFU/g of log reduction.

The rhizome of Homalomena josefii P.C. Boyce and S.Y. Wong is commonly used in Borneo folk medicine. It is potent in treating stomach aches, headaches, and snake bites. The lack of information about their biological activities leads to an investigation to determine its antibacterial and antioxidant activities of H. josefii rhizome extract. This study aimed to ascertain antibacterial activity in opposition to foodborne pathogens namely Bacillus megaterium ATCC14581, Bacillus pumilus ATCC14884, Proteus mirabilis ATCC21100 and Klebsiella pneumoniae ATCC13773 and its antioxidant properties of H. josefii rhizome extracts. Extraction of dried powdered H. josefii rhizome was carried out through the maceration method using methanol as a solvent to produce crude extracts. The crude extracts were then tested for antibacterial and antioxidant activities. The antibacterial activity was conducted in terms of disc diffusion assay (DDA), minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and time-kill assay. All were performed following the Clinical and Laboratory Standard Institute (CLSI) procedure to ensure liable results. Total phenolic compounds (TPC) and 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) scavenging assay were used to stimulate the antioxidant activity. The results show that DDA inhibition zone of the methanolic extracts ranged from 9.00±0.58 to 10.00±0.00 mm. The extract was able to inhibit the growth of all tested bacteria with MICs value ranging from 0.31 to 5.00 mg/mL. Meanwhile, the extract is able to kill all tested bacteria with MBC values that range from 0.63 to 5.00 mg/mL. Time-kill assay curve analysis results showed that the extract was able to completely kill the bacterial growth at 4 × MIC for 4 hours. The total phenolic compound (TPC) of methanolic extract was 435.138 mg GAE/g and IC50 of the extract was 11.809 μg/mL. In conclusion, the methanolic extract of H. josefii rhizome extract exhibited antibacterial and antioxidant activities, thus it can further be developed as a natural preservative.

Bacillus cereus spores are capable of surviving the harsh environment and more often, they cause great concern to the dairy industry. The current research was conducted to study the effect of temperature on germination and growth of B. cereus spores in UHT chocolate milk; the study was carried out at 8°C, 25°C and 35°C over a span of seven days. The results showed that no growth was observed at 8°C. At 25°C, a rapid increase in growth was observed as early as Day 1, from an initial count of ten spores to 4.01 log10 CFU/mL. Meanwhile, at 35°C, the growth on Day 1 was more rapid in which the count promptly increased to 8.07 log10 CFU/mL. Analysis of graph trend showed that the number of vegetative cells decreased while the number of spores increased with incubation time due to nutrients exhaustion. This study fills up the data gap towards understanding the possible issues that might arise in the actual scenario and at the same time, suggests a suitable approach to minimize infection risk caused by B. cereus spores.

Salmonella is the well-recognized foodborne pathogen leading the most research and surveillance attention especially from government agencies as well as in food industry.In Malaysia, Salmonella is one of the main bacteria which monitored by the National Laboratory Surveillance System.Previously, standard culture methods have always been employed by many laboratories for Salmonella detection in Food Surveillance Programs.However, more advanced detection methods will be needed to improve the sensitivity and specificity of Salmonella identification.In this review, Salmonella detection methods including conventional and recent advances in molecular-based methods will be discussed.

F U L L P A P E R Influence of food composition type on the microwave heating time in relation to the inactivation of Salmonella enterica serovar Enteritidis and Shiga-toxigenic Escherichia coli (STEC) O157 New, C.Y., Abdul Rahman, R., Mohammed, A.S., Ubong, A., Chang, W.S., Thung, T.Y., Tan, C.W., Lee, E., Tang, J-Y-H. and Son, R. Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia Department of Food Technology, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia Department of Microbiology and Biotechnology, Faculty of Science, Federal University Dutse, Jigawa State, Nigeria Faculty of Bioresources and Food Industry, Universiti Sultan Zainal Abidin, 22200 Besut, Terengganu Food Safety and Food Integrity, Institute of Tropical Agriculture and Food Security, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

Peanuts are widely consumed as the main ingredient in many local dishes in Malaysia. However, the tropical climate in Malaysia (high temperature and humidity) favours the growth of fungi from Aspergillus section Flavi, especially during storage. Most of the species from this section, such as A. flavus, A. parasiticus and A. nomius, are natural producers of aflatoxins. Precise identification of local isolates and information regarding their ability to produce aflatoxins are very important to evaluate the safety of food marketed in Malaysia. Therefore, this study aimed to identify and characterize the aflatoxigenic and non-aflatoxigenic strains of Aspergillus section Flavi in peanuts and peanut-based products. A polyphasic approach, consisting of morphological and chemical characterizations was applied to 128 isolates originating from raw peanuts and peanut-based products. On the basis of morphological characters, 127 positively identified as Aspergillus flavus, and the other as A. nomius. Chemical characterization revealed six chemotype profiles which indicates diversity of toxigenic potential. About 58.6%, 68.5%, and 100% of the isolates are positive for aflatoxins, cyclopiazonic acid and aspergillic acid productions respectively. The majority of the isolates originating from raw peanut samples (64.8%) were aflatoxigenic, while those from peanut-based products were less toxigenic (39.1%). The precise identification of these species may help in developing control strategies for aflatoxigenic fungi and aflatoxin contamination in peanuts, especially during storage. These findings also highlight the possibility of the co-occurrence of other toxins, which could increase the potential toxic effects of peanuts.

High demand for milk has been observed amongst the Malaysian public.Hence, research in milk is essential to assure food safety in milk consumption.This study evaluated the quality of locally-produced milk and present of bacterial hazards in cow and goat milk.A total of 120 milk samples including thirty raw cow milk, thirty pasteurized cow milk, thirty raw goat milk and thirty pasteurised goat milk were collected from dairy farms, delivery milkman, marts and markets in Selangor, Malaysia.The bacteriological quality of milk was evaluated for the presence of Escherichia coli, mesophilic, and coliform bacteria.An acceptable standard limit of < 1 x 10 5 CFU/mL for the total bacterial count was used to indicate good quality of milk.Overall, all type of milk exceeded 100,000 CFU/mL.The pasteurized raw goat milk showed the highest (7.16 log 10 CFU/mL) in total plate count while the pasteurized cow milk recorded as the lowest (5.38 log 10 CFU/mL) in total plate count.Approximately half of the milk samples were contaminated with coliform bacteria and a proportion has exceeded the acceptable limit of 50 CFU/mL.The presence of E. coli was detected in over 44% of the samples.Milk contaminated with the pathogenic E. coli can cause self-limited, watery to bloody diarrhea including severe diseases like haemolytic uremic syndrome (HUS).Hence, it is important to ensure the quality of milk for public health safety.

Shiga-toxin producing Escherichia coli bacteria are well known to be the pathogenic bacteria that cause traveler diarrhea.E. coli O157: H7 from the group of Shiga-toxin producing E. coli cause even severe infection which can lead to fatality for humans.In this study, local beef and Indian buffalo were selected to determine the presence of Shiga-toxin producing E. coli and E. coli O157: H7 using Most Probable Number-Polymerase Chain Reaction (MPN-PCR) method.Among 108 samples, eight (7.41%) samples from local beef and Indian buffalo were detected a positive on E. coli O157: H7 while thirteen (12.04%) samples were detected positive for Shiga-toxin producing E. coli gene.Out of 108 samples, eleven isolates of E. coli O157: H7 were successfully isolated in order to carry out the antibiotic susceptibility test.Shiga-toxin producing E. coli isolates were found susceptible to ceftazidime (100%), moxifloxacin (83.33%), sulphamethoxazole (66.67%), ampicillin (50%), amoxycillin (50%), ciprofloxacin (50%), erythromycin (33.33%) and penicillin G (33.33%).E. coli O157: H7 isolates were susceptible toward erythromycin (100%), ceftazidime (100%), ciprofloxacin (100%) and moxifloxacin (100%), sulphamethoxazole (60%), ampicillin (20%), amoxycillin (20%), and penicillin G (0%).The safety of both local beef and Indian buffalo was challenged by the presence of both Shiga-toxin producing E. coli and E. coli O157: H7.Better and safer ways of removing the pathogen from local beef and Indian buffalo should be researched more deeply.

*Thung, T.Y., Lee, E., Premarathne, J.M.K.J.K., Nurzafirah, M., Kuan, C.H., Elexson, N., Tan, C.W., Malcolm, T.T.H., New, C.Y., Ramzi, O.S.B., Nuzul, N.J., Noor Azira, A.M., Ungku Fatimah, U.Z.A. and Son, R. Department of Food Science, Faculty of Food Safety and Technology, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia Food Safety and Food Integrity, Institute of Tropical Agriculture and Food Security, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia Department of Livestock and Avian Science, Faculty of Livestock, Fisheries and Nutrition, Wayamba University of Sri Lanka, Makandara, 60170 Gonawila, Sri Lanka Department of Diagnostic and Allied Science, Faculty of Health and Life Science, Management and Science University, 40100 Shah Alam, Selangor, Malaysia Department of Agricultural and Food Science, Faculty of Science, Universiti Tunku Abdul Rahman, 31900 Kampar, Perak, Malaysia Department of Biology Molecule, Faculty of Resource Science and Technology, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia Department of Food Service and Management, Faculty of Food Safety and Technology, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia

Tan, C.W., Noor Hazirah, M.Z., Shu’aibu, I., New, C.Y., Malcolm, T.T.H., Thung, T.Y., Lee, E., Wendy, R.D. Nuzul, N.J., Noor Azira, A.M., Ungku Fatimah, U.Z.A., Rukayadi, Y., Rinai, R. and Son, R. Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia Department of Food Service and Management, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM Serdang, Serdang, Selangor Darul Ehsan, Malaysia Food Safety and Food Integrity, Institute of Tropical Agriculture and Food Security (ITAFoS), Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

This goal of this study was to investigate the presence of Vibrio cholerae in street food, namely satar and otak-otak, using Loop-Mediated Isothermal Amplification (LAMP), multiplex Polymerase Chain Reaction (mPCR) and conventional plating on Thiosulphate Citrate Bile-Salt Sucrose (TCBS) agar methods.A total of 78 satar and 35 otak-otak were purchased from different districts of Terengganu (Besut, Setiu, Kuala Terengganu and Kemaman).V. cholerae was found in satar with LAMP (10.3%), mPCR (10.3%) and plating (0%).No V. cholerae was found in otak-otak using the three methods.This might be due to V. cholerae able to survive in satar after grilling due to its thickness which may contribute to undercooking.This study concluded that low presence of V. cholerae in satar and otak-otak can be detected by molecular methods but not the conventional plating method.LAMP assay is a useful tool for rapid detection of pathogens in food due to its simplicity, highly sensitive and visual interpretation capability.Though the prevalence of V. cholerae was low in the samples, proper handling of this food will help in reducing the risk of acquiring infection from V. cholerae in contaminated samples.

F U L L P A P E R Simultaneous multiplex Polymerase Chain Reaction detection of Salmonella spp., Escherichia coli O157, Vibrio parahaemolyticus, Vibrio cholerae, Listeria monocytogenes and Campylobacter spp. Ling, S., Noramirah, R., Abidatul, A.A., Nurfarhanah, N.M.J., Noor-Azira, A.M., Jambari, N.N., Ungku Fatimah, U.Z.A., New, C.Y. and Son, R. Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia Food Safety and Food Integrity, Institute of Tropical Agriculture, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia Department of Food Service and Management, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

The safety level of microwaved foods remains at vague as this subject was less addressed scientifically.A study was initiated to address the matter by investigating on the survivability of Salmonella and Shiga-toxigenic Escherichia coli (STEC) O157 in microwave heated ready-to-eat (RTE) foods using the Most Probable Number coupled Polymerase Chain Reaction (MPN-PCR) technique.A total of 329 samples of various ready-to-eat (RTE) convenience meals were collected around Wilayah Persekutuan Kuala Lumpur and Selangor regions.Salmonella was positively identified in 66 samples (20.1%, <3.0-11000 MPN/g) while 86 samples (26.1%, <3.0 ->11000 MPN/g) were positive for E. coli.Out of the 66 positive Salmonella samples, S. enterica serovar Typhimurium was identified in 6 samples (1.8%, <3.0-62.0MPN/g) and S. enterica serovar Enteritidis was identified in 13 samples (4.0%, <3.0-270 MPN/g).On the other hand, 17 out of the 86 positive E. coli samples were identified as positive STEC O157 (5.2%, 3.0-930 MPN/g).The results signified the high possibility of the pathogens' survival in RTE foods due to uneven heat distribution, resulting in the presence of cold spots which supports the growth of the pathogens, as well as the microwave reheating time and lack consumers' knowledge on the microwave oven.The risk of contracting foodborne illness from the consumption of survived pathogens in microwave heated RTE food was estimated using the @RISK® Version 7.5 (Palisade, USA).The outcome indicated a moderate to high rate of foodborne illness incidence which indicated the need to create the awareness on the safety of microwaved foods and provide proper microwaving guidelines to mitigate the risk.