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Escherichia coli O157:H7 is a major food-borne pathogen that has resulted in numerous outbreaks around the world. Widespread distribution of the organism in various ecological niches impedes the control measures. This study aimed to detect and quantify E. coli O157:H7 in beef sold in wet markets and hypermarkets in Malaysia and to determine the risk of E. coli O157:H7 infection linked to consumption of beef. The rfbO157 and flicH7 primers targeted on somatic antigen (O157) and flagellar antigen (H7) respectively of E. coli O157:H7 was used for the MPN-PCR method. A total of 99 beef samples were collected from local wet markets and hypermarkets. The highest E. coli O157:H7 contamination rate was observed in beef samples collected from wet markets (89.50%), whereas the contamination rate in hyper market A and B were compratively low (35.35 and 20% respectively). However, the microbial load was highest in the beef samples from hypermarket A (1100 MPN/g) while E. coli O157:H7 bacterial load in beef samples from hypermarket B and wet market ranged from 3 to 93 MPN/g and 3 to 240 MPN/g, respectively. Using the Quantitative Microbial Risk Assessment (QMRA) approach the risk was estimated incorporating the findings of the prevalence study and predictions based on home storage, cooking and consumption patterns. Three different exposure pathways were investigated to estimate the risk associated with contaminated beef and Monte Carlo simulation was used to determine the level of uncertainty. The developed model predicated that consumption of contaminated beef can be accountable for 1.83E+06 E. coli O157:H7 cases per year in Malaysia. The reliability of the model, data gaps and further research needs, is discussed. Through continuous improvement Quantitative Microbial Risk Assessment provides valuable insight into controlling and prevention strategies.

Cross contamination is one of the most important contributing factors in foodborne illness originating in household environments. The objective of this research was to determine the transfer between naturally contaminated chicken liver and leg to cutting board, hand glove, knife and cucumber, during slicing. The microorganism tested was Campylobacter jejuni and the results showed that the pathogen transferred to all utensils, at different transfer rate, despite the low level of the naturally contaminating pathogen. With unknown concentration bacteria in the naturally contaminated samples, a proportion of the utensils were still contaminated with C. jejuni and not surprisingly, when the sample were contaminated with higher concentrations of the pathogen, a higher proportion of the utensils had detectable C. jejuni cells present, though in many cases cross contamination seems to be a random event. Transfer of the naturally contaminating C. jejuni from the chicken liver and leg to the utensils were

Foodborne pathogens have become a constant threat to the consumer and food industry. Reduce efficacy of antibiotics with emergence of resistant bacteria has limited the opportunities for controlling pathogenic bacteria in food commodities and treating foodborne infections. Bacteriophages can be a promising alternative for alleviate the risk of transmitting pathogenic bacteria via food commodities. Therefore, this research was conducted to find distribution of bacteriophages in diverse niches in order to identify suitable sources for isolating bacteriophages to use controlling foodborne pathogens. Firstly bacterial strains were screened for lysogenic and selected suitable host bacterial strains were used for isolating and determining bacteriophage titer in fresh raw food and environmental samples. Eighteen different lytic bacteriophages effective against Campylobacter, S. aureus, L. monocytogenes and E. coli were isolated from this study. Bacteriophages titer was determined within range of 102 to 1010 PFU/mL and bacteriophages were most frequently isolated from chicken (60%) samples. The isolated bacteriophages could be potential candidates for controlling foodborne diseases. [ABSTRACT FROM AUTHOR]

Vibrio parahaemolyticus is a halophilic Gram-negative bacterium that is considered among gastrointestinal pathogens. Thirty isolates were tested for their susceptibility using 14 different antibiotics. One V. parahaemolyticus isolate was resistant to 10 antibiotics (cefotaxime, ceftazidime, tetracycline, amikacin, ciprofloxacin, levofloxacin, of loxacin, ampicillin, amoxicillin-calv-acid, and cefepime). The V. parahaemolyticus isolates were resistant to ampicillin (90%), amoxicillin-clavulanic acid (63.3%), cefotaxime (60%), ceftazidime (46.7%), cefepime (50%), tetracycline (36.6%), and amikacin (26.7%). However, the isolates were highly susceptible to imipenem (100%), and piperacillin and gentamicin (96.7%). Approximately 55% of the isolates showed a multiple antibiotic resistance (MAR) index of >0.2, thereby indicating the high risk of sources where these isolates originated. The occurrence of MAR asserted the importance of determining drug susceptibility and monitoring the antimicrobial resistance profile to improve and ensure food safety and public health. [ABSTRACT FROM AUTHOR]

Vibrio parahaemolyticus is a main foodborne disease in seafood and generally seafood is easily deteriorates in quality of color and flavor. In this study, clove (Syzygium aromaticum) extract shows potent antibacterial activity against growth of antibiotics resistant Vibrio parahaemolyticus on seafood samples (cockles and shrimps). Vibrio parahaemolyticus was artificial contaminates on the samples with 106 CFU/ml. The samples were treated with different concentration of cloves extract with 10 mg/ml which are 0.5%, 5% and 10% concentration from methanol food grade extraction in 0 hr, 5 min, 10 min, 15 min, 30 min, 60 min and 120 min. Tab water and deionized water were selected as a negative control. As a result, the amount of 10 % cloves managed to mitigates the number of V. parahaemolyticus on seafood samples in 5 minutes and 15 min on both samples. Therefore, our results signify the fact that cloves can be apply as natural sanitizer which could meet consumer demands for safe and traditionally consumed either raw without any undesirable effect when applied in the seafood system industries. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes (L. monocytogenes) is an important foodborne pathogen which can cause foodborne listeriosis with high mortality rates especially in susceptible population groups such as pregnant women, elderly and immunocompromised individuals. The biosafety level of L. monocytogenes in chicken offal has becomes a great concern as chicken offal is a cheap source of protein and it is often served as side dishes in South East Asian countries. In Malaysia, the consumption of chicken offal has almost doubled from 5 g per capita per day in the early 1980s to 9 g per capita per day in 2009. In this study, risk assessment was conducted to estimate the risk of acquiring listeriosis from consumption of chicken offal in Malaysia. A microbial survey on the prevalence and concentration of L. monocytogenes in chicken offal were carried out in Selangor, Malaysia over a one-year period (November 2010 to October 2011). It was assumed that there were no seasonal changes in the prevalence and consumption pattern all year round. Assuming that 5.6 million people in Selangor, Malaysia consume a single serving (125 g) of chicken offal per week, it is estimated that in a year there could be 0.61 cases and 1.98 x 10-4 cases of listeriosis per 100,000 population of pregnant woman and immunocompromised individual, respectively. However, the potential for getting listeriosis among the healthy population was very low, only 1.39 x 10-8 cases per 100,000 population. This study demonstrated risk assessment model not only used as a tool to estimate the risk of acquiring illness but it can influence public health surveillance and providing data in setting approriate level of protection. [ABSTRACT FROM AUTHOR]

Vibrio parahaemolyticus is recognized as a frequent causal agent of human gastroenteritis due to the consumption of raw, undercooked or mishandled seafood in many Asian countries. The number of V. parahaemolyticus cases reported is on the rise, and this becomes a concern to the Asian countries as seafood is favoured by Asians. This study aimed to detect and quantify V. parahaemolyticus in raw oysters and to determine the risk associated with the consumption of raw oysters. A total of 30 oyster samples were collected and analysed in this study. MPN- PCR and MPN-Plating methods were employed and carried out concurrently to determine the prevalence of V. parahaemolyticus in raw oysters. The results showed that the prevalence of total V. parahaemolyticus in oysters was 50.00% (15/30) where the MPN/g range was < 3 -- > 11000 MPN/g for MPN-PCR method, and 40.00% (12/30) where the MPN/g range was < 3 -- 4300 MPN/g for MPN-Plating method. MPN-PCR method was able to estimate the level of virulence (tdh+ and trh+) V. parahaemolyticus in the raw oysters where 10.00% (3/30) of samples were identified to be in a range of 3 -- 30 MPN/g. A microbial risk assessment was conducted based on the enumeration data obtained from MPN-PCR method using @risk. The probability of illness annually was 1.76 X 10-6 with a prediction of 31 cases to occur with respect to the exposed Malaysian population, while the rate per 100,000 people was estimated to be at 0.104. In addition, the antibiogram of V. parahaemolyticus was determined using Kirby Bauer Disk Diffusion Test and the results indicated that the isolates were highly resistant towards Bacitracin (100.00%), Vancomycin (100.00%) and were least resistant to Chloramphenicol (8.70%). The MAR index of the isolates ranged from 0.17 to 0.50. In accordance with the results from this study, the consumption of raw oysters is a risk factor for V. parahaemolyticus infection and proactive actions should be taken to reduce the risk of the pathogen in order to improve public health. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes (L. monocytogenes) is a gram positive food-borne pathogen that is able to form biofilm on food factory surfaces. Formation of biofilm makes the bacteria much more resistance to environmental stresses such as disinfectant. The extracellular polymeric matrix (biofilm structure) which is mostly comprised of sticky extracellular polysaccharides (EPS) and proteins can protect bacteria in a harsh condition. The efficiency of four disinfectants on removing L. monocytogenes biofilm was investigated. Five concentration levels (100, 50, 25, 12.5, and 6.25%) of disinfectants were tested. In the microtitre assay, the optical density at 595 nm CV-OD595 value, was used to measure the amount of remained biofilm after 24 h. Results showed that disinfectants did not have significant effect on removing L. monocytogenes biofilm. Formation of L. monocytogenes biofilm significantly decreased the efficiency of disinfectants. Biofilm produced by strain number 9 showed higher resistance to disinfectant. Low concentrations (<50%) of disinfectants did not show significant effect on removing L. monocytogenes biofilm. [ABSTRACT FROM AUTHOR]

Bacillus cereus (B. cereus) isolates are toxigenic and can cause food poisoning. Cooked rice is a potentially hazardous food, especially in tropical countries. The aim of this study was to determine the prevalence of B. cereus and B. thuringiensis in raw and cooked rice marketed in Selangor, Malaysia. A combination of Most Probable Number - Polymerase Chain Reaction (MPN-PCR) method was used to detect gyrB gene in B. cereus and B. thuringiensis. Five local varieties of raw rice samples were negative for B. thuringiensis but all (100%) were positive for B. cereus. A total of 115 cooked rice samples (nasi lemak, nasi briyani, nasi ayam and nasi putih) were studied for the presence of B. cereus and B. thuringiensis. Nasi ayam was found to have the highest prevalence (100%) of B. cereus compared to nasi putih (76.2%) and nasi lemak (70.4%). Nasi briyani had the lowest prevalence (50%) of B. cereus. The frequencies of B. thuringiensis were found to be 10, 30 and 35.2 % in nasi putih and nasi ayam, nasi briyani and nasi lemak, respectively. Occurrence of B. cereus and B. thuringiensis in the samples ranged from < 3 to 1100 MPN/g in different samples. Maximum number of B. cereus was observed in nasi lemak, nasi briyani and nasi putih ( > 1100 MPN/g) while nasi ayam showed less contamination (460 MPN/g) with B. cereus which was significantly different (P < 0.05 ) from others. The number of B. thuringiensis in nasi lemak, nasi briyani, nasi putih and nasi ayam were found to be >1100, 93, 9.2 and 3.6 MPN/g, respectively. [ABSTRACT FROM AUTHOR]

This study was undertaken to characterize the antibiotic resistance and randomly amplified polymorphic DNA (RAPD) profiles of Vibrio parahaemolyticus isolates from raw vegetable samples. A total of 46 isolates of V. parahaemolyticus recovered from raw vegetables samples and were confirmed by PCR were analyzed in this study. Most of the isolates were resistant to nalidixic acid (93.48%) and were the least resistant towards imipinem (4.35%). The MAR index results also demonstrated high individual and multiple resistances to antibiotics among the isolates. From the RAPD analysis, the size for RAPD fragments generated ranged from 250 bp to 1,500 bp, with most of the strains contained three major gene fragments of 350, 1,000 and 1,350 bp. The RAPD profiles revealed a high level of DNA sequence diversity within the isolates. Antibiotic resistance and RAPD proved to be effective tools in characterizing and differentiating the V. parahaemolyticus strains. [ABSTRACT FROM AUTHOR]

A total of 49 isolates of V. parahaemolyticus and 8 isolates of V. cholerae isolated from freshwater fish of patin (Pangasius hypopthalmus) and red tilapia (Oreochromis sp.) were purchased from different retail level in Selangor, Malaysia. All of the isolates showed a multiple resistances towards all 15 antibiotics tested. Some of the isolates show a high resistance to different antibiotics including bacitracin, vancomycin, tetracycline, furazididone, cephalothin and erythromycin. However, both species was susceptible towards imipenem. Overall antibiotics resistance patterns of all isolates were resistant from 2 to 14 resistance patterns with multiple antibiotic resistance (MAR) index ranging from 0.13 to 0.93 respectively. As the results obtained in the dendrogram produced from both species had indicates that these antibiotics were intensively used whether in the aquaculture farm through feeds during culture or at the hatchery production of seed. Thus, this study will provides an essential information of the MAR index and also the clustering analysis in order to determine the biosafety of Vibrio spp. in freshwater aquaculture fish sold at different retail level in Malaysia. [ABSTRACT FROM AUTHOR]

In this study, the prevalence of Vibrio spp. and V. cholerae in flavored drinks and fruit juices sold at hawker stalls and in restaurants were determined using the most probable number (MPN) method and polymerase chain reaction (PCR) assay. One hundred and twenty drinks samples of four types of drinks (Iced Milk Rose Syrup, n=25; Iced Milk Corn Syrup, n=20; Apple Juice, n=37 and Iced Carrot Milk, n=28) were collected at two different settings; mainly hawker stalls and restaurants. Upon analysis, the prevalence of Vibrio spp. and V. cholerae in fruit juices and flavored drinks for hawker stalls were 93.3% and 36.7%, respectively whereas for the restaurants, were 93.3% and 20% respectively. Overall, the prevalence of Vibrio spp. and V. cholerae in all the drink samples were 93.3% and 28.3% respectively, highlighting the needs for the improvement of the hygiene and sanitation practice in the settings studied. [ABSTRACT FROM AUTHOR]

Little is known on the biosafety level of Vibrio spp. in freshwater fish in Malaysia. The purpose of this study was to investigate the prevalence and concentration of Vibrio spp. and V. parahaemolyticus in freshwater fish using the Most Probable Number-Polymerase Chain Reaction (MPN-PCR) method. The study was conducted on 150 samples from two types of freshwater fish commonly sold at hypermarkets, i.e. Pangasius hypophthalmus (catfish) and Oreochromis sp. (red tilapia). Sampling was done on the flesh, intestinal tract and gills of each fish. The prevalence of Vibrio spp. and V. parahaemolyticus was found to be 98.67% and 24% respectively with higher percentages detected in samples from the gills followed by the intestinal tract and flesh. Vibrio spp. was detected in almost all red tilapia and catfish samples. V. parahaemolyticus was detected in 25% of the catfish samples compared to 22.6% of red tilapia fish. The density of Vibrio spp. and V. parahaemolyticus in the samples ranged from 0 to 1.1x107 MPN/g. Although the maximum value was 1.1x107 MPN/g, most samples had microbial loads ranging from 0 to >104 MPN/g. The outcome on the biosafety assessment of Vibrio spp. and V. parahaemolyticus in freshwater fish indicates another potential source of food safety issues to consumers. [ABSTRACT FROM AUTHOR]

Several Norovirus cases due to consumption of green onions have been reported during recent years but reports on red onions are not found. Onions are one of the major tastes in Malaysian food which are sometimes consuming raw especially the green onion. Viral contamination in onions can occur due to planting condition and not properly prepared food. This situation can pose the human health risk. A method was developed to detect the Norovirus that might present on different type of onions. In this study, 60 samples were collected from local market. Elution by Tryptose Phosphate Glycine broth and concentration steps using negatively charge filter were applied to enhance the detection of virus in food due to low copies of virus on food surface. The viral RNA was extracted using Qiagen Rneasy Mini kit before further detection using One-step RT-PCR. The total incidence of Norovirus in green onion and red onion was 13.33% (4/30) and 3.33 % (1/30) respectively. This is the first report of the detection of Norovirus in red and green onions in Malaysia. Based on the results, it is concluded that this method is reliable to detect Norovirus on onions and vegetables surface and hence can be applied in the laboratories for routine or food borne outbreak investigation. [ABSTRACT FROM AUTHOR]

This study aims to determine the frequency and density of potentially pathogenic Vibrio parahaemolyticus, defined as those possessing thermostable-direct hemolysin (tdh) and/or tdh-related hemolysin (trh) genes, in raw salad vegetables at retail level in Selangor, Malaysia. A combination of Most Probable Number - Polymerase Chain Reaction (MPN-PCR) method was applied to detect the presence of tdh and/or trh gene-possessing V. parahaemolyticus and to enumerate their density in the samples. A total of 276 samples of vegetables commonly eaten raw in Malaysia (Cabbage = 30; Carrot = 31; Cucumber = 28; Four winged bean = 26; Indian pennywort = 17; Japanese parsley = 21; Lettuce = 16; Long bean = 32; Sweet potato = 29; Tomato = 38; Wild cosmos = 8) were analyzed. The samples were purchased from two supermarkets (A and B) and two wet markets (C and D). With the MPN-PCR technique, about 12.0% of the samples were positive for the presence of V. parahaemolyticus tdh-positive, with maximum densities of up to 39 MPN/g. The total frequency of V. parahaemolyticus trh-positive in the samples was 10.1%, with maximum concentration 15 MPN/g. V. parahaemolyticus tdh-positive was most prevalent in samples from Wet Market C (20.78%) and also in vegetable type Oenanthe stolonifera (Japanese parsley) with 19.0%, while V. parahaemolyticus trh-positive was predominant in samples from Wet Market D (16.7%) and was most frequent in both Oenanthe stolonifera (Japanese parsley) and Cucumis sativus (Cucumber) with 14.3% prevalence for each type. The results highlighted the fact that raw vegetables could be contaminated with virulent V. parahaemolyticus and could act as a transmission route, thus poses risk to consumers from the consumption of raw vegetables. To the author's knowledge, this is the first assessment of V. parahaemolyticus carrying tdh and trh genes in raw vegetables from retail outlets in Malaysia. [ABSTRACT FROM AUTHOR]

To gain insight into the microbiological safety of food products routinely traded across international borders in Southeast Asian countries, beef imported from Malaysia to southern Thailand was examined for contamination with Escherichia coli O157 and its subsequent spread into the imported areas. We screened 31 samples exported from Malaysia and 36 domestic Thai samples. Isolation methods including an O157 antigen-targeted immunomagnetic separation technique, screening on CHROMagar O157 medium, and serotype confirmation of E. coli isolates by specific agglutination tests were employed. Fourteen strains of E. coli O157:H7 were isolated from eight Malaysian samples (25.8%) and six strains from four Thai samples (11.1%). These strains were of the stx1- stx2+ eae+ genotype except one Malaysian strain which was of the stx1- stx2- eae+ genotype. All 19 O157:H7 strains possessing the stx2 gene produced little or no Stx2 (reversed passive latex agglutination titer ≤ 4). Of the 19 strains, five Malaysian (38.5%) and two Thai (33.3%) strains exhibited resistance to a set of antibiotics. Finally, the results of two DNA fingerprinting analyses (O157 IS-printing targeted to IS629 and pulsed-field gel electrophoresis, PFGE) of the O157:H7 strains possessing the stx2 gene, indicated that the Malaysian and Thai strains are closely related. Therefore, E. coli O157:H7 might be transferred from Malaysia to southern Thailand through beef trade. [ABSTRACT FROM AUTHOR]