Displaying publications 1 - 20 of 33 in total

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  1. Nakaguchi Y
    Trop Med Health, 2013 Sep;41(3):95-102.
    PMID: 24155650 DOI: 10.2149/tmh.2011-06
    Infections by virulent strains of Vibrio parahaemolyticus are frequently reported in Southeast Asia. This is due to the frequent seafood contamination by virulent strains. In this study conducted from 2008 to 2011, seafood like fish, shrimp, squid, crab, and molluscan shellfish were purchased from provinces in Thailand and three Southeast Asian countries and examined for the prevalence of three genetic markers of V. parahaemolyticus (species-specific gene: toxR gene, virulence genes: tdh and trh genes). An enrichment culture of seafood was examined for these markers using PCR methods. Molluscan shellfish showed a high frequency of contamination in Thailand. The shellfish harvested from the Gulf of Thailand were significantly more contaminated with virulence genes than those from the Andaman Sea. The seafood purchased from three Southeast Asian countries was positive for the three markers of V. parahaemolytcus at differing frequencies. The virulence markers (tdh and trh markers) were frequently detected in molluscan shellfish from Vietnam (17.9 and 8.0%, respectively), Malaysia (11.1 and 16.7%), and Indonesia (9.1 and 13.6%). These data suggest that the molluscan shellfish sold in Southeast Asian markets are highly contaminated with virulent strains of V. parahaemolyticus.
  2. Pui CF, Wong WC, Chai LC, Lee HY, Noorlis A, Zainazor TC, et al.
    Trop Med Health, 2011 Mar;39(1):9-15.
    PMID: 22028607 DOI: 10.2149/tmh.2010-20
    Salmonellosis outbreaks involving typhoid fever and human gastroenteritis are important diseases in tropical countries where hygienic conditions are often not maintained. A rapid and sensitive method to detect Salmonella spp., Salmonella Typhi and Salmonella Typhimurium is needed to improve control and surveillance of typhoid fever and Salmonella gastroenteritis. Our objective was the concurrent detection and differentiation of these food-borne pathogens using a multiplex PCR. We therefore designed and optimized a multiplex PCR using three specific PCR primer pairs for the simultaneous detection of these pathogens. The concentration of each of the primer pairs, magnesium chloride concentration, and primer annealing temperature were optimized before verification of the specificity of the primer pairs. The target genes produced amplicons at 429 bp, 300 bp and 620 bp which were shown to be 100% specific to each target bacterium, Salmonella spp., Salmonella Typhi and Salmonella Typhimurium, respectively.
  3. Kuan CH, Goh SG, Loo YY, Chang WS, Lye YL, Puspanadan S, et al.
    Poult Sci, 2013 Jun;92(6):1664-9.
    PMID: 23687164 DOI: 10.3382/ps.2012-02974
    A total of 216 chicken offal samples (chicken liver = 72; chicken heart = 72; chicken gizzard = 72) from wet markets and hypermarkets in Selangor, Malaysia, were examined for the presence and density of Listeria monocytogenes by using a combination of the most probable number and PCR method. The prevalence of L. monocytogenes in 216 chicken offal samples examined was 26.39%, and among the positive samples, the chicken gizzard showed the highest percentage at 33.33% compared with chicken liver (25.00%) and chicken heart (20.83%). The microbial load of L. monocytogenes in chicken offal samples ranged from <3 to 93.0 most probable number per gram. The presence of L. monocytogenes in chicken offal samples may indicate that chicken offal can act as a possible vehicle for the occurrence of foodborne listeriosis. Hence, there is a need to investigate the biosafety level of chicken offal in Malaysia.
  4. Goh SG, Kuan CH, Loo YY, Chang WS, Lye YL, Soopna P, et al.
    Poult Sci, 2012 Oct;91(10):2686-90.
    PMID: 22991558
    This study aimed to determine the prevalence Listeria monocytogenes in raw chicken meat samples at hypermarkets and wet markets. Chicken drumsticks, breasts, and thighs were randomly selected. The most probable number (MPN) PCR method was used to quantify the L. monocytogenes in the samples. Listeria monocytogenes was detected in 20% of the samples. Occurrence of L. monocytogenes was highest in breast (42.03%) followed by drumstick (11.27%) and thigh (7.14%). Samples from hypermarkets showed higher occurrence (25.71%) of L. monocytogenes compared with wet markets (14.29%). The density of L. monocytogenes found in samples ranged from <3.0 to 16 MPN•g(-1). The presence of L. monocytogenes in raw chicken meat is unwanted but unpreventable. Thus, further research on the processing method to reduce and eliminate this kind of bacteria in chicken meat before consumption is necessary. The presence of L. monocytogenes in chicken samples suggests the importance of this pathogen in chicken. Thus, more study is needed to find ways to eliminate this pathogen from poultry.
  5. Thung TY, Mahyudin NA, Basri DF, Wan Mohamed Radzi CW, Nakaguchi Y, Nishibuchi M, et al.
    Poult Sci, 2016 Aug 01;95(8):1888-93.
    PMID: 27118863 DOI: 10.3382/ps/pew144
    Salmonellosis is one of the major food-borne diseases in many countries. This study was carried out to determine the occurrence of Salmonella spp., Salmonella Enteritidis, and Salmonella Typhimurium in raw chicken meat from wet markets and hypermarkets in Selangor, as well as to determine the antibiotic susceptibility profile of S. Enteritidis and S. Typhimurium. The most probable number (MPN) in combination with multiplex polymerase chain reaction (mPCR) method was used to quantify the Salmonella spp., S. Enteritidis, and S. Typhimurium in the samples. The occurrence of Salmonella spp., S. Enteritidis, and S. Typhimurium in 120 chicken meat samples were 20.80%, 6.70%, and 2.50%, respectively with estimated quantity varying from <3 to 15 MPN/g. The antibiogram testing revealed differential multi-drug resistance among S. Enteritidis and S. Typhimurium isolates. All the isolates were resistance to erythromycin, penicillin, and vancomycin whereas sensitivity was recorded for Amoxicillin/Clavulanic acid, Gentamicin, Tetracycline, and Trimethoprim. Our findings demonstrated that the retail chicken meat could be a source of multiple antimicrobial-resistance Salmonella and may constitute a public health concern in Malaysia.
  6. Koitabashi T, Vuddhakul V, Radu S, Morigaki T, Asai N, Nakaguchi Y, et al.
    Microbiol. Immunol., 2006;50(2):135-48.
    PMID: 16490932
    Nine Escherichia coli O157: H7/- strains isolated primarily from non-clinical sources in Thailand and Japan carried the stx(2) gene but did not produce Stx2 toxin in a reversed passive latex agglutination (RPLA) assay. A strain (EDL933) bearing a stx(2) phage (933W) was compared to a strain (Thai-12) that was Stx2-negative but contained the stx(2) gene. To study the lack of Stx2 production, the Thai-12 stx(2) gene and its upstream nucleotide sequence were analyzed. The Thai-12 stx(2) coding region was intact and Stx2 was expressed from a cloned stx(2) gene using a plasmid vector and detected using RPLA. A lacZ fusion analysis found the Thai-12 stx(2) promoter non-functional. Because the stx(2) gene is downstream of the late promoter in the stx(2) phage genome, the antitermination activity of Q protein is essential for strong stx(2) transcription. Thai-12 had the q gene highly homologous to that of Phi21 phage but not to the 933W phage. High-level expression of exogenous q genes demonstrated Q antitermination activity was weak in Thai-12. Replication of stx(2) phage was not observed in Stx2-negative strains. The q-stx(2) gene sequence of Thai-12 was well conserved in all Stx2-negative strains. A PCR assay to detect the Thai-12 q-stx(2) sequence demonstrated that 30% of O157 strains from marketed Malaysian beef carried this sequence and they produced little or no Stx2. These results suggest that stx(2)-positive O157 strains that produce little or no Stx2 may be widely distributed in the Asian environment.
  7. Tang JY, Nishibuchi M, Nakaguchi Y, Ghazali FM, Saleha AA, Son R
    Lett Appl Microbiol, 2011 Jun;52(6):581-8.
    PMID: 21375548 DOI: 10.1111/j.1472-765X.2011.03039.x
    We quantified Campylobacter jejuni transferred from naturally contaminated raw chicken fillets and skins to similar cooked chicken parts via standard rubberwood (RW) and polyethylene cutting boards (PE).
  8. Chai LC, Ghazali FM, Bakar FA, Lee HY, Suhaimi LR, Talib SA, et al.
    J Microbiol Biotechnol, 2009 Nov;19(11):1415-20.
    PMID: 19996696
    The aim of the present study was to examine the prevalence of thermophilic Campylobacter spp. (Campylobacter jejuni and Campylobacter coli) in soil, poultry manure, irrigation water, and freshly harvested vegetables from vegetable farms in Malaysia. C. jejuni was detected in 30.4% and 2.7% of the soil samples, 57.1% and 0% of the manure samples, and 18.8% and 3% of the vegetable samples from farm A and farm B, respectively, when using the MPNPCR method. Campylobacter spp. was not found in any of the irrigation water samples tested. Therefore, the present results indicate that the aged manure used by farm A was more contaminated than the composted manure used by farm B. Mostly, the leafy and root vegetables were contaminated. C. coli was not detected in any of the samples tested in the current study. Both farms tested in this study were found to be contaminated by campylobacters, thereby posing a potential risk for raw vegetable consumption in Malaysia. The present results also provide baseline data on Campylobacter contamination at the farm level.
  9. Tunung R, Margaret S, Jeyaletchumi P, Chai LC, Tuan Zainazor TC, Ghazali FM, et al.
    J Microbiol Biotechnol, 2010 Feb;20(2):391-6.
    PMID: 20208446
    The purpose of this study was to investigate the biosafety of Vibrio parahaemolyticus in raw salad vegetables at wet market and supermarket in Malaysia. A combination of Most Probable Number - Polymerase Chain Reaction (MPN-PCR) method was applied to detect the presence of V. parahaemolyticus and to enumerate their density in the food samples. The study analyzed 276 samples of common vegetables eaten raw in Malaysia (Wild cosmos = 8; Japanese parsley = 21; Cabbage = 30; Lettuce = 16; Indian pennywort = 17; Carrot = 31; Sweet potato = 29; Tomato = 38; Cucumber = 28; Four winged bean = 26; Long bean = 32). The samples were purchased from two supermarkets (A and B) and two wet markets (C and D). The occurrence of V. parahaemolyticus detected was 20.65%, with higher frequency of V. parahaemolyticus in vegetables obtained from wet markets (Wet market C = 27.27%Wet Market D = 32.05%) compared to supermarkets (Supermarket A = 1.64%; Supermarket B = 16.67%). V. parahaemolyticus was most prevalent in Indian pennywort (41.18%). The density of V. parahaemolyticus in all the samples ranged from <3 up to >2400 MPN/g, mostly <3 MPN/g concentration. Raw vegetables from wet markets contained higher levels of V. parahaemolyticus compared to supermarkets. V. parahaemolyticus were present in raw vegetables although in low numbers. The results suggest that raw vegetables act as a transmission route for V. parahaemolyticus. This study will be the first biosafety assessment of V. parahaemolyticus in raw vegetables in Malaysia.
  10. Chai LC, Lee HY, Ghazali FM, Abu Bakar F, Malakar PK, Nishibuchi M, et al.
    J Food Prot, 2008 Dec;71(12):2448-52.
    PMID: 19244897
    Campylobacter jejuni was found to occur at high prevalence in the raw salad vegetables examined. Previous reports describe cross-contamination involving meat; here we investigated the occurrence of cross-contamination and decontamination events in the domestic kitchen via C. jejuni-contaminated vegetables during salad preparation. This is the first report concerning quantitative cross-contamination and decontamination involving naturally contaminated produce. The study was designed to simulate the real preparation of salad in a household kitchen, starting with washing the vegetables in tap water, then cutting the vegetables on a cutting board, followed by slicing cucumber and blanching (heating in hot water) the vegetables in 85 degrees C water. Vegetables naturally contaminated with C. jejuni were used throughout the simulation to attain realistic quantitative data. The mean of the percent transfer rates for C. jejuni from vegetable to wash water was 30.1 to 38.2%; from wash water to cucumber, it was 26.3 to 47.2%; from vegetables to cutting board, it was 1.6 to 10.3%; and from cutting board to cucumber, it was 22.6 to 73.3%. The data suggest the wash water and plastic cutting board as potential risk factors in C. jejuni transmission to consumers. Washing of the vegetables with tap water caused a 0.4-log reduction of C. jejuni attached to the vegetables (most probable number/gram), while rapid blanching reduced the number of C. jejuni organisms to an undetectable level.
  11. Wong, W.C., Pui, C.F., Tunung, R., Cheah, Y.K., Nakaguchi, Y., Nishibuchi, M., et al.
    MyJurnal
    A total of 112 burger patties (35 beef burger patties, 39 chicken burger patties and 38 fish burger patties) which are commercially available at retail level were investigated for the presence and number of Listeria monocytogenes. These samples were analyzed using MPN-PCR method and conventional culturing methods. L. monocytogenes was detected in 33.3% of chicken burger patties, 22.9% of beef patties, and 10.5% of fish patty samples. From all contaminated raw burger patties, the estimated count of L. monocytogenes was ranged from 3 to 75 MPN/g. The results suggest that burger act as a potential source of listeriosis if the contaminated burger patty is consumed without adequate cooking. The risk associated with consumption of these samples was found to be high particularly for processed food at retail level in Malaysia. Therefore, food manufacturers play an important role in monitoring the manufacturing process and conduct a periodical surveillance on microbiological quality assessment on the processing plants. Besides, there is a need to increase awareness of consumers and food handlers to practice proper cooking of the burger patties before the point of consumption, to reduce the risk of listeria infection.
  12. Noor Hidayah, M.S., Tuan Zainazor, C., Pui, C.F., Noorlis, A., Noor Eliza, M.R., Naziehah, M.D., et al.
    MyJurnal
    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.
  13. Jeyaletchumi, P., Tunung, R., Margaret, S.P., Son, R., Ghazali, F.M., Cheah, Y.K., et al.
    MyJurnal
    The aim of this study was to assess the most probable number-polymerase chain reaction (MPNPCR) technique for detection of Listeria monocytogenes in salad vegetables in comparison with reference EN ISO 11290-2 and Food Drug Administration Bacteriological Analytical Manual method using artificial and naturally contaminated samples. Based on recovery of L. monocytogenes from artificially contaminated samples, MPN-PCR showed a moderate correlation (R=0.67) between spiking concentration and microbial levels which was better than the FDA-BAM method (R=0.642) and ISO 11290-2:1998 method (R=0.655). With naturally contaminated samples, it was found that L. monocytogenes was detected in 25% of the vegetable samples using MPN-PCR; 15% of the samples by the FDA-BAM method and 8% of samples using ISO 11290-2:1998 method. Overall, MPN-PCR was found to be a rapid and reliable method that could facilitate the enumeration of L. monocytogenes in vegetables.
  14. Sandra, A., Afsah-Hejri, L., Tunung, R., Tuan Zainazor, T. C., Tang, J. Y. H., Ghazali, F. M., et al.
    MyJurnal
    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. In this research combination of Most Probable Number - Polymerase Chain Reaction (MPN-PCR) 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. The range of B. cereus and B. thuringiensis in the samples was 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.
  15. Tunung, R., Jeyaletchumi, P., Noorlis, A., Tang, Y.H., Sandra, A., Ghazali, F.M., et al.
    MyJurnal
    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.
  16. Afsah-Hejri, L., Rukayadi, Y., Fouladynezhad, N., Son, R., Nakaguchi, Y., Nishibuchi, M.
    MyJurnal
    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 (
  17. Chai, L.C., Fatimah, A.B., Ghazali, F.M., Lee, H.Y., Tunung, R., Shamsinar, A.T., et al.
    MyJurnal
    Antibiotic resistance in campylobacter is an emerging global public health problem after MRSA and VRE. Fluoroquinolone and macrolide resistance have been found to be more common in this world leading foodborne pathogen. A total of fifty-six isolates of Campylobacter jejuni obtained from raw vegetables
    which are consumed as ulam (salad) in Malaysia, were tested with 12 antibiotics used clinically and
    agriculturally. The resistance was determined using the disk diffusion method. Results were determined
    by hierarchic numerical methods to cluster strains and antibiotics according to similarity profiles. Fifty
    five C. jejuni isolates from different isolation sites were all clustered together into ten groups. This indicates that the commodities (raw salad vegetables/ulam) where the isolates originated might share a similar source of cross-contamination along the production route. All antibiotics tested correlated and there were four groupings reflecting their mode of actions. Generally, C. jejuni isolates were found to be highly resistant to erythromycin (91.1%) and tetracycline (85.7%). Both agents are popular antibiotics used clinically to treat bacterial infections. On the other hand, the C. jejuni isolates showed high percentage (80.4%) of resistance towards enrofloxacin, an extensively used antimicrobial agent in agriculture practices. This study showed that C. jejuni isolates were highly multi-resistance to as many as 10 antibiotics. Therefore, in terms of biosafety, the presence of antibiotic resistance strains in the food chain has raised concerns that the treatment of human infections will be compromised.
  18. Tang, J.Y.H., Carlson, J., Mohamad Ghazali, F., Saleha, A.A., Nishibuchi, M., Nakaguchi, Y., et al.
    MyJurnal
    The present study aimed to provide an insight of C. jejuni ATCC 33560 phenotype profiles (carbon sources and sensitivity to osmolytes and pH) using Phenotypic MicroArray (PM) system in response to optimal and suboptimal temperature. C. jejuni ATCC 33560 showed utilization carbon sources from amino acids and carboxylates but not from sugars. C. jejuni ATCC 33560 is sensitive to NaCl at 2% and above but showed survival in a wide range of food preservatives (sodium lactate, sodium phosphate, sodium benzoate, ammonium sulphate and sodium nitrate). When incubated at suboptimal temperature, no phenotype loss was observed in carbon source plates. Phenotype loss of C. jejuni ATCC 33560 was observed in sodium chloride (1%), sodium sulphate (2-3%), sodium formate (1%), sodium lactate (7-12%), sodium phosphate pH7 (100mM and 200mM), ammonium sulphate pH8 (50mM), sodium nitrate (60mM, 80mM and 100mM), sodium nitrite (10mM), and growth in pH5. The phenotypic profile from present study will provide a better insight related to survival of C. jejuni ATCC 33560.
  19. New, C.Y., Kantilal, H.K., Tan, M.T.H., Nakaguchi, Y., Nishibuchi, M., Son, R.
    MyJurnal
    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.
  20. Elexson, N., Rukayadi, Y., Nakaguchi, Y., Nishibuchi, M.
    MyJurnal
    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.
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