Effects of green tea extract on reducing Vibrio parahaemolyticus and increasing shelf life of oyster meats

This study investigated effects of tea extract on growth of pathogenic Vibrio parahaemolyticus and potential utilization in post-harvest treatment to extend shelf life of Pacific oysters (Crassostrea gigas). Longjing Tea, which exhibited strong bactericidal activity against V. parahaemolyticus, was selected to use in this study. Tea extract containing equal or higher than 4.6 g/l total phenolic contents (TPC) as gallic acid equivalents (GAE) determined by Folin-Ciocalteau method could reduce a mixture of five clinical V. parahaemolyticus strains in tryptic soy broth plus 1.5% NaCl from 4.5 log CFU/ml to non-detectable level (<1 log CFU/ml) within 8 h. A treatment of shucked oysters with tea extract containing 9.1 g/l TPC as GAE for 2 h at 23 ± 1 °C with oyster/tea extract ratio of approximate 0.9 g/ml resulted in greater (p < 0.05) V. parahaemolyticus reductions (0.8 log MPN/g) compared to controls (0.2 log MPN/g). The following shelf life study indicated that green tea treatment at oyster/tea extract ratio of approximate 0.7 g/ml could enhance reducing V. parahaemolyticus while retarding the growth of total bacteria in oysters during 5 ± 1 °C storage. Therefore, green tea might be utilized as a natural antimicrobial agent to inactivate V. parahaemolyticus in oysters and extend the shelf life during refrigeration storage.     

Prevalence and antimicrobial susceptibility of Vibrio parahaemolyticus isolated from retail shellfish in Shanghai

Vibrio parahaemolyticus is a marine and estuarine bacterium that poses the greatest threat to human health worldwide. It has been the leading bacterial cause of seafood-borne illness. This study investigated the prevalence and drug resistance of V. parahaemolyticus isolated from retail shellfish in Shanghai. A total of 140 shellfish samples were collected from February 2014 to February 2015. The occurrence of V. parahaemolyticus in shellfish was 34.3%, which has increased compared to previous reports. In addition, discernible differences of total presumptive V. parahaemolyticus counts (TPVPC) were also observed in shellfish between market A and B. The results from PCR assays indicated that thermostable direct hemolysin (tdh) gene was positive in two isolates (2.1%), and the thermostable direct hemolysin-related hemolysin (trh) gene was not detected in all isolates. Antibiotic resistance profiles of those isolates were as follows: ampicillin (87.5%), cephazolin (31.3%), cephalothin (6.3%), amoxicillin/clavulanic acid (6.3%), piperacillin (6.3%), and amikacin (3.2%). Thirty-three out of 96 isolates were resistant to two or more antimicrobial agents. It is suggested that V. parahaemolyticus in retail shellfish could be a potential risk to consumers in Shanghai.     

Prevalence,pathogenicity, and serotypes of Vibrio parahaemolyticus in shrimp from Chinese retail markets

Vibrio parahaemolyticus is one of the most common foodborne pathogens in Asian countries. V. parahaemolyticus contamination of retail shrimp in China has been reported previously, and such contaminated shrimp have been linked to outbreaks of foodborne diseases. However, to date, the prevalence of V. parahaemolyticus in retail shrimp in China has not been determined. This study aimed at identifying the prevalence of V. parahaemolyticus in shrimps in Chinese retail market. From May 2012 to April 2013, a total of 273 shrimp samples was obtained from retail markets in 16 provinces (19 cities) of China. V. parahaemolyticus was detected in 103 (37.7%) of 273 samples by the most probable number method. Bacterial densities were less than 100 MPN/g in 95.1% (98/103) of the samples. Five trh-positive isolates were identified from 247 isolates, and none of the isolates were tdh-positive. In multiplex PCR-based O-antigen serotyping of these 247 pathogenic isolates, all O-antigen serotypes, except O9, were detected, and serotype O2 was found to be the most prevalent (detected in 82 isolates). This is the first report on V. parahaemolyticus prevalence in retail shrimp in China, and the findings of this study can be used for microbiological risk assessment of shrimp in China.     

Prevalence and antimicrobial susceptibility of Vibrio species related to food safety isolated from shrimp cultured at inland ponds in Thailand

Inland farming of marine shrimp species is a relatively recent development, in response to increasing demand for the products. This study aimed to investigate the prevalence and antimicrobial resistance of potentially pathogenic Vibrio species in shrimps cultured at inland ponds with low salinity in Thailand. Of 16 shrimp samples (white-leg shrimps and black-tiger shrimps) from ponds with water temperatures from 29 to 32 °C, and salinities from 1 to 5 parts per trillion, 15 contained Vibrio cholerae (densities: 62–252,000 MPN/g). Vibrio parahaemolyticus was present in six samples (370–6,300,000 MPN/g), and Vibrio vulnificus was present in two samples (16–1300 MPN/g). V. parahaemolyticus has higher affinity for non-native white-leg shrimp than for native black-tiger shrimp. The minimum inhibitory concentrations were then measured for 12 common antimicrobial agents. Resistance to ampicillin and oxytetracycline (8% and 2% of the isolates) was documented in V. cholerae isolates, resistance to ampicillin and oxytetracycline (72% and 3%) was found in V. parahaemolyticus isolates, and resistance to nalidixic acid (20%) was detected in V. vulnificus isolates. β-lactamase and tetracycline resistance genes were detected in the resistant Vibrio isolates. The oxytetracycline-resistance phenotype was eliminated by plasmid curing, suggesting that the resistance was related to R-plasmids. Our results show that shrimps cultured even in low salinity ponds are potentially contaminated with pathogenic Vibrio species, and that shrimp are a vehicle for transfer of species with antimicrobial resistance genes to the public.     

The efficacy of grape seed extract,citric acid and lactic acid on the inactivation of Vibrio parahaemolyticus in shucked oysters

The efficacy of grape seed extract (GE), citric acid (CA) or lactic acid (LA) on the inactivation of Vibrio parahaemolyticus in shucked oysters was studied. The minimum inhibitory concentration (MIC) of GE, CA or LA against V. parahaemolyticus in TSB-1% NaCl was also determined. The shucked oysters were artificially inoculated with V. parahaemolyticus, the inoculated shucked oysters (25 g) were then dipped in solution of GE (0.0, 10.0, 20.0, 50.0, 100, 200, 300 and 500 mg mL⁻¹), CA (0.0, 5.0, 10.0, 15.0, 20.0, 50.0, 100, 200 and 300 mg mL⁻¹) or LA (0.0, 1.0, 5.0, 10.0, 15.0, 20.0, 50.0, 100 and 150 mg mL⁻¹) for 10 min. The population of V. parahaemolyticus in shucked oysters was determined. The MICs of GE, CA or LA against V. parahaemolyticus were 10.0, 5.0 or 1.0 mg mL⁻¹, respectively. A 500, 300 or 150 mg mL⁻¹ GE, CA or LA solutions were needed to reduce the population of V. parahaemolyticus to below the detection level (1.0 log g⁻¹) in shucked oysters.     

Pomegranate peel (Punica granatum L) extract and Chinese gall (Galla chinensis) extract inhibit Vibrio parahaemolyticus and Listeria monocytogenes on cooked shrimp and raw tuna

Vibrio parahaemolyticus and Listeria monocytogenes are bacterial pathogens associated with raw or ready-to-eat seafood products. Many compounds extracted from plant material have shown promise for inhibiting bacterial pathogens when applied to some foods. In this study, aqueous methanol extracts from pomegranate peel (Punica granatum L.) and Chinese gallnut (Galla chinensis) were tested against V. parahaemolyticus and L. monocytogenes on cooked shrimp and raw homogenized tuna. The extracts were applied to the shrimp by soaking for 2 min (5 mg/ml). The extracts (1.7 mg ml) were added to homogenized tuna and stirred. The antimicrobial assay on V. parahaemolyticus was conducted at 12 °C, and the assay on tuna was conducted at both 4° and 12 °C. Both Chinese gall and pomegranate peel extracts significantly inhibited the growth of V. parahaemolyticus in both shrimp and tuna. Only Chinese gall extract significantly inhibited growth of L. monocytogenes. Overall V. parahaemolyticus was more sensitive to both plant extracts compared with L. monocytogenes. Both plant extracts had stronger antimicrobial activity on shrimp compared with the tuna. Neither extract completely inhibited the growth of V. parahaemolyticus or L. monocytogenes.     

An insight into the inhibitory activity of dihydromyricetin against Vibrio parahaemolyticus

The objective of this study is to investigate the antibacterial activity of dihydromyricetin (DMY) against Vibrio parahaemolyticus. The dilution method indicated that the minimum inhibitory concentration (MIC) of DMY against V. parahaemolyticus was at 0.625 mg/mL. The inhibitory effects of DMY against V. parahaemolyticus was further studied by analyzing cell morphology, cell injury, cell permeability, cell surface hydrophobicity (CSH) and antibacterial rate. The results showed bacterium cells are completely inactivated in a higher concentration (10 MIC). DMY treatment also lead to an increase in cell membrane permeability, cell injury as well as CSH. A good correlation between antibacterial rate and CSH was also observed. These findings indicated DMY could be used as a new alternative natural antibacterial agent for control pathogen growth in aquatic food.     

PMA based real-time fluorescent LAMP for detection of Vibrio parahaemolyticus in viable but nonculturable state

Vibrio parahaemolyticus is a crucial foodborne pathogen. The viable but non-culturable (VBNC) state of V. parahaemolyticus cannot be detected by traditional culture methods. The objective of this study was to develop and evaluate a method that combines propidium monoazide (PMA) treatment with real-time fluorescent loop-mediated isothermal amplification (RF-LAMP) to detect and quantify VBNC cells of V. parahaemolyticus. Different states of cells were treated with PMA in dark for 5 min and subsequently exposed to a 650 W halogen lamp for 5 min. The cells were collected and DNA was amplified by RF-LAMP. The primers which targeted six distinct regions in the tlh gene of V. parahaemolyticus were used for the PMA-RF-LAMP assay. The results indicated that the treatment with 4 μg/mL of PMA and a further exposure to light for 5 min was suitable for PMA-RF-LAMP to distinguish viable cells from dead cells of V. parahaemolyticus. The developed assay exhibited remarkably high specificity, sensitivity and rapidity, without any cross reaction with the tested non-V. parahaemolyticus strains. There was good linear correlation between Ct values and log copy/mL in the range of 6.8–6.8 × 10⁷ copy/mL, with the reaction endpoints less than 30 cycles (30 min). It could detect as low as 14 copy/g of V. parahaemolyticus in spiked seafood samples (pomfret, shrimp, scallop, oyster and salted fish) without any interference from food matrices, dead cells and other bacteria.     

In vitro control of multiplication of some food-associated bacteria by thyme, rosemary and sage isolates

Antibacterial activity of thyme, rosemary and sage isolates obtained by supercritical fluid extraction and hydrodistillation was investigated on Geobacillus stearotermophillus, Bacillus cereus, Bacillus subtilis var. niger, Enterococcus faecium, Salmonella enteritidis and Escherichia coli strains. Bacillus species were the most susceptible to all tested isolates. The thyme isolates showed the strongest antibacterial activity against tested bacteria with MIC values of 40-640 μg/ml, followed by rosemary (MIC = 320-1280 μg/ml) and sage (MIC = 160-2560 μg/ml) isolates. Therefore, the antibacterial activity of the most abundant components found in the thyme isolates, thymol, p-cymene and their mixture was investigated as well. The thyme isolates, especially supercritical extract, showed stronger antibacterial activity against Bacillus strains compared to the single components and their mixture, which indicated synergetic effect of the other components. Results of this study indicated thyme as a valuable source of natural antibacterial agents and supercritical fluid extraction as an efficient isolation method.     

Antimicrobial activity of rhamnolipids against Listeria monocytogenes and their synergistic interaction with nisin

The rhamnolipids (RL) are biodegradable biosurfactants which have low toxicity and surface active properties that can be useful for food processing industries. The objective of this study was to evaluate the antimicrobial potential of rhamnolipids against Listeria monocytogenes. Susceptibility tests were performed by the minimal inhibitory concentration (MIC) using the micro-broth dilution technique. The MIC values varied from 78.1 μg/mL to 2500 μg/mL with the 2500 μg/mL being the predominant value. Among the 32 tested cultures, 90.6% were susceptible to RL. Results showed that the rhamnolipid activity was primarily bacteriostatic. The interaction of rhamnolipid with nisin was also investigated. The combined effect of nisin and RL was evaluated against two wild-type isolates of L. monocytogenes, L12 sensitive to RL (MIC 156.2 μg/mL) and L17 less sensitive to RL (2500 μg/mL). The FIC indexes for the isolates were 0.18 and 0.078 for L12 and L17 respectively, indicating a strong synergistic effect. The survival curve of isolates L12 and L17 showed that the combination between nisin and RL was bactericidal at lower concentration than for the individual antimicrobials. For the L12 isolate 78.1 μg/mL of RL and 160 IU/mL of nisin eliminated the population after 30 min of incubation. The combination of 156.2 μg/mL of RL and 320 IU/mL of nisin reduced completely L17 population after 2 h of incubation. Rhamnolipids showed antimicrobial activity against L. monocytogenes and presented a synergistic effect when combined with nisin.     

In vitro enhancement of antibiotic susceptibility of drug resistant Escherichia coli by cinnamaldehyde

Cinnamaldehyde is a natural antimicrobial compound that has been found to damage the cytoplasmic membrane, inhibit septum development and cause cell elongation, as well as induce oxidative stress in Escherichia coli. Thus, cinnamaldehyde may be of value as a natural compound to enhance susceptibility of drug resistant E. coli to antibiotics in livestock production. This study examined the ability of cinnamaldehyde to increase the susceptibility of E. coli to antibiotics using the checkerboard method. Interactions between the antimicrobials were characterized using fractional inhibitory concentration (FIC) values. All tested E. coli strains were resistant to erythromycin and bacitracin but were susceptible to penicillin G and ampicillin. Strains 8WT, ATCC 23739 and 02:0627 were resistant to novobiocin and the latter two strains were also resistant to tetracycline. Cinnamaldehyde synergistically increased the susceptibility of all E. coli strains to erythromycin (FIC ≤ 0.5). Another synergistic effect between tetracycline and cinnamaldehyde was observed when tested against E. coli ATCC 23739 (FIC = 0.3). Cinnamaldehyde synergistically and additively reduced the MIC of novobiocin when tested against ATCC 23739 and 02:0627 (FIC ≤ 0.5) or 8WT (FIC = 1), respectively, suggesting that E. coli strains may respond differently to challenge by the cinnamaldehyde-novobiocin combination. With all strains, cinnamaldehyde was not effective at reducing the MIC value of bacitracin. Findings of this study suggest that cinnamaldehyde may be used in combination with several antibiotics to enhance susceptibility of drug resistant E. coli.     

In vitro assessment of synthetic phenolic antioxidants for inhibition of foodborne Staphylococcus aureus,Bacillus cereus and Pseudomonas fluorescens

Minimal inhibitory concentrations (MICs) of seven synthetic phenolic compounds, five commonly used as antioxidants (TBHQ, BHA, BHT, propyl gallate and octyl gallate) and two as antimicrobials (propyl 4-hydroxybenzoate and n-heptyl 4-hydroxybenzoate) were assessed against several strains of two Gram-positive (Staphylococcus aureus and Bacillus cereus) and one Gram-negative (Pseudomonas fluorescens) bacteria, by using a standardized microdilution assay (ISO 20776-1, 2006). Octyl gallate was the most effective compound against the three genera/species of bacteria considered simultaneously (with the exception of four strains of B. cereus, which were resistant for this compound) with MIC values (≤100 μg/ml) lower than the concentrations usually used as antioxidants. TBHQ and n-heptyl 4-hydroxybenzoate were also effective in the control of S. aureus at very low concentrations (MIC of 3.1 μg/ml and 12.5 μg/ml, respectively). Propyl 4-hydroxybenzoate was the most inhibitory phenolic compound against all strains of B. cereus and both tested parabens (propyl- and heptyl-) were not effective for P. fluorescens (MIC > 1600 μg/ml). B. cereus was the bacterial genera that showed more intra-species variation, distinguishing two clearly groups of sensitivity among the strains to octyl gallate and n-heptyl 4-hydroxybenzoate (“sensitive” with mean MICs of 42.8 and 4.2 μg/ml, respectively; and “resistant” with MICs >1600 and >800 μg/ml, respectively). According to all that, octyl gallate would be an interesting phenolic compound for the food industry, not only because of its recognized antioxidant properties but also because of their effectivity as antimicrobial against S. aureus, B. cereus and P. fluorescens.     

Production and characterization of a monoclonal antibody against recombinant thermolabile hemolysin and its application to screen for Vibrio parahaemolyticus contamination in raw seafood

Thermolabile hemolysin gene (tlh) is regarded as a species-specific marker for Vibrio parahaemolyticus. To assess the utility of the tlh gene product, thermolabile hemolysin (TLH), as a marker to screen for V. parahaemolyticus-contamination in raw seafood, we generated a monoclonal antibody (MAb) against recombinant TLH and established with an enzyme-linked immunosorbent assay using the MAb (TLH-ELISA). TLH-ELISA testing of broth cultures for 78 V. parahaemolyticus strains showed positive results all around. In contrast, most broth cultures of 53 non-V. parahaemolyticus species tested yielded negative results.We devised a screening method using TLH-ELISA to check for low-level contamination of V. parahaemolyticus in raw seafood within 24 h and evaluated its ability. In testing of V. parahaemolyticus-spiked raw seafood, results suggested that our screening method can detect 100 most-probable-number (MPN) of V. parahaemolyticus/g. Further, on testing 119 commercial raw seafood samples with our screening method, 117 samples were determined to contain less than 100 MPN of V. parahaemolyticus/g. All of the 117 samples were also estimated by the MPN method to contain less than 100 MPN of V. parahaemolyticus/g. Taken together, these results suggest that our screening method using TLH-ELISA is useful to check for low-level (<100 MPN/g) of V. parahaemolyticus in raw seafood.     

Selective turn-on fluorescence detection of Vibrio parahaemolyticus in food based on charge-transfer between CdSe/ZnS quantum dots and gold nanoparticles

Vibrio parahaemolyticus (V. parahaemolyticus) is one of the most common food-borne pathogens found along the east coast of China, Japan, and several southeast Asian countries, where marine foods are frequently consumed. Thus it is of great interest to establish a fast, sensitive and robust analytical assay for V. parahaemolyticus detection. Herein, we report a V. parahaemolyticus-specific egg yolk antibody (IgY), extracted from the egg yolks of immunized hens with inactivated V. parahaemolyticus bacteria. By immobilizing IgY onto the surface of gold nanoparticles (AuNPs) through electrostatic self-assembly, a detection system was developed based on charge-transfer quenching fluorescence, which contains core/shell CdSe/ZnS quantum dots (QDs), IgY-AuNPs and free AuNPs. In this strategy, the IgY-AuNPs were induced to aggregate quickly in the presence of target V. parahaemolyticus, resulting in a rapid fluorescence response of the QDs. More importantly, the convenient turn-on fluorescent detection system exhibited excellent selectivity over the other bacteria. Furthermore, this facile one-step method can detect V. parahaemolyticus at low concentrations of 10 cfu/mL without pre-enrichment and can be applicable for the detection of V. parahaemolyticus in real food samples. With the advantage of simplicity, rapidness, sensitivity, and specificity, this proposed approach shows promise as a successful application of V. parahaemolyticus in bioanalysis.     

Comparison of Vibrio parahaemolyticus isolates from aquatic products and clinical by antibiotic susceptibility,virulence, and molecular characterisation

Vibrio parahaemolyticus is a common foodborne pathogen found in aquatic products and represents a major threat to human health worldwide. Though not all this bacteria were harmful to human beings, the pathogenic V. parahaemolyticus always harbors either tdh (the thermostable direct hemolysin) or trh (TDH-related hemolysin) gene, or both. Additionally, the extensive use of antibiotics has been shown to be a contributing factor to the increasing incidence of antimicrobial-resistant strains. In this study, thirty-one clinical isolates were examined and compared with 95 (38.0%) aquatic product isolates (fishes, n = 28; shrimps, n = 67) collected from 250 samples in Guangdong, China. All isolates were studied by antibiotic susceptibility analysis, tdh and trh genes detection, serotyping and molecular typing (ERIC-PCR). The antimicrobial resistance patterns of these aquatic product isolates to 12 antimicrobial agents revealed that most of the isolates were resistant to streptomycin (90.53%). The isolates were also resistant to follow by ampicillin (33.68%) and cephalothin (30.53%). For clinical isolates, they were resistant to streptomycin (93.55%), ampicillin (87.10%), and cefazolin (64.52%). All isolates showed no resistance to azitromycin, chloramphenicol, ciprofloxacin, or nalidixic acid. The clinical isolates were positive for tdh (100%) and trh gene (77.42%), with ratios of only 2.11% and 28.42%, respectively in the aquatic product isolates. Serotyping detected shown that the isolates contained O1, O2, O3, O4, and O11, with the O3 serotype being the most common among the clinical isolates (48.39%), while the O2 (41.05%) makes the maximum proportion on aquatic product isolates. ERIC-PCR results demonstrated the isolates (n = 126) were classified into eight clusters, revealing genetic variation and relatedness between clinical and aquatic product isolates. This study provided a foundation for understanding the distinction between aquatic product and clinical isolates and yielded basic information for achieving food safety through control of V. parahaemolyticus contamination.     

Susceptibility of Clostridium perfringens to antimicrobials produced by lactic acid bacteria: Reuterin and nisin

The effectiveness as antimicrobials of lactic acid bacteria produced compounds reuterin and nisin was assessed against vegetative cells and spores of Clostridium perfringens isolates (from ovine milk obtained in farms with diarrheic lambs) and C. perfringens CECT 486 (type A toxin producer). We also tested the inhibitory effect of lysozyme and sodium nitrite on Clostridium. Minimal inhibitory concentrations (MIC) of antimicrobials were determined in modified RCM (mRCM) and milk by using a broth microdilution method, after 7 d at 37 °C under anaerobic conditions. The sensitivity of C. perfringens to the tested antimicrobials was strain and culture medium-dependent. In general, vegetative cells exhibited higher sensitivity than spores. Reuterin (MIC values 2.03–16.25 mM) inhibited the growth of vegetative cells and the outgrowth of spores of all tested C. perfringens, both in mRCM and milk, with higher resistance in milk. Nisin (MIC values 0.78–12.5 μg/ml) was also effective against vegetative cells and spores of tested C. perfringens in both culture media. However, lysozyme (up to 400 μg/ml) did not control the growth of any of the tested Clostridium. Sodium nitrite only inhibited the outgrowth of spores of two C. perfringens isolates at the maximum concentration assayed (300 μg/ml) exclusively in mRCM medium. These results suggest that reuterin and nisin have the potential to control the growth of C. perfringens, and might help to ensure safety at different stages of the food chain. Future studies in food/feed products would be necessary to further corroborate this hypothesis.     

Combining basic electrolyzed water pretreatment and mild heat greatly enhanced the efficacy of acidic electrolyzed water against Vibrio parahaemolyticus on shrimp

The objective of this study was to investigate the efficacy of acidic electrolyzed water (AEW) against Vibrio parahaemolyticus on shrimp. The shrimp was initially inoculated with V. parahaemolyticus(7-8 log CFU/g), and treated with AEW (AEW1 containing 51 mg/L of chlorine or AEW2 containing 78 mg/L of chlorine) or organic acids (2% AA and 2%LA) for 1 min or 5 min under different treated conditions. The effect of AEW was better than that of organic acids, the number of survival V. parahaemolyticus cells on shrimp was reduced by 0.9 log CFU/g after treatment for 5 min with AEW without vibration, while 1.0 log CFU/g bacteria cells reduced with vibration. No significant difference (p > 0.05) was observed between AEW and organic acids in the bactericidal activity with or without vibration. The effective order of temperatures on bactericidal activities of AEW was 50 °C > 20 °C > 4 °C, and a 3.1 log CFU/g reduction of V. parahaemolyticus cells on shrimp was detected with treatment of AEW at 50 °C. Mild heat greatly enhanced efficacy of electrolyzed water against V. parahaemolyticus. Basic electrolyzed water (BEW) (50 °C) pretreatment combined with AEW (50 °C) treatment remarkably reduced bacterial cells by 5.4 log CFU/g on shrimp after treatment for 5 min. There was a significant change in physicochemical properties (pH, ORP, ACC) of AEW, after it was used to wash shrimp (P < 0.05). This study suggests that BEW (50 °C) pretreatment followed by AEW (50 °C) treatment could be a possible method to effectively control V. parahaemolyticus contamination on shrimp.     

Epidemiology of foodborne disease outbreaks caused by Vibrio parahaemolyticus during 2010–2014 in Zhejiang Province,China

Vibrio parahaemolyticus is one of the most commonly reported species causing foodborne diseases in China, to gain understanding of epidemiology characteristics of V. parahaemolyticus outbreaks in Zhejiang Province, China, we summarized data on V. parahaemolyticus outbreaks from 2010 to 2014 in Zhejiang Province reported to China National Foodborne Diseases Surveillance Network. A food item was implicated if V. parahaemolyticus was isolated from food or based on epidemiologic evidence. From 2010 to 2014, 71 outbreaks due to V. parahaemolyticus were reported, resulting in 933 illnesses and 117 hospitalizations without death. The median annual incidence rate of confirmed V. parahaemolyticus outbreaks reported was 2.75 per million population (rang, 1.11–5.60). About 90.1% (64/71) of outbreaks caused by V. parahaemolyticus occurred during May and October. Aquatic products (27 outbreaks, 38.0%), meat and meat products (9 outbreaks, 12.7%), plant-based foods (6 outbreaks, 8.4%), mixed foods (5 outbreaks, 7.0%) were the most commonly implicated foods. Outbreaks most frequently occurred in restaurants (50.7%), private residents (21.1%), and cafeteria (12.7%). Cross contamination (39.4%), improper cooking (21.1%), improper storage (16.9%) were main reasons. To prevent V. parahaemolyticus outbreaks caused by cross contamination, improper cooking and improper storage in high-temperature seasons, regulations for seafood safety from the production stage to the consumption stage should be strengthened.     

Application of grape seed extract in depuration for decontaminating Vibrio parahaemolyticus in Pacific oysters (Crassostrea gigas)

This study investigated the potential application of grape seed extract (GSE) in depuration to increase its efficacy in reducing Vibrio parahaemolyticus populations in Pacific oysters (Crassostrea gigas). Pacific oysters were inoculated with five clinical strains of V. parahaemolyticus to 10⁴⁻⁵ MPN/g and depurated in UV-irradiated artificial seawater (ASW) containing GSE at a level of 1.0% (1.8 mg/mL total phenolic contents as gallic acid equivalents) or 1.5% (3.1 mg/mL total phenolic contents as gallic acid equivalents) at 12.5 °C for up to 5 days. Changes of V. parahaemolyticus populations in oysters during depuration were analyzed every 24 h using the three-tube most probable number (MPN) method. The populations of V. parahaemolyticus in inoculated oysters decreased by 3.06 and 3.71 log MPN/g, respectively, after 4 and 5 days of depuration in ASW at 12.5 °C. However, populations of V. parahaemolyticus in oysters were reduced by 3.01 and 4.18 log MPN/g after 2 days of depuration at 12.5 °C in ASW containing 1.0 and 1.5% GSE, respectively. Further studies confirmed that depuration using ASW containing 1.5% GSE with 3.1 mg/mL total phenolic contents as gallic acid equivalents at 12.5 °C for two days was capable of achieving >3.52 log MPN/g reductions of V. parahaemolyticus in Pacific oysters.     

Seasonal prevalence of Vibrio species in retail shrimps with an emphasis on Vibrio parahaemolyticus

Seasonal prevalence of Vibrio species in shrimp samples from retail outlets in the South-western part of Iran was studied. A total of 300 samples were analyzed (75 samples in each season). Special attention was paid to the prevalence of total and pathogenic Vibrio parahaemolyticus. All the TCBS isolates were first identified to the genus level with PCR and then identified to the species level using a battery of biochemical reactions and tests. To investigate the pathogenicity of the isolated V. parahaemolyticus, multiplex PCR (tl, tdh and trh genes) was performed. Vibrios were detected during the whole investigation period, depending on the sampling season. They were detected in 18.6% of the winter samples, 64% of the spring samples, 70.6% of the summer samples and 41.3% of the autumn samples. Vibrio calviensis and Vibrio alginolyticus were dominant in samples of different seasons, with the average prevalence of 18.6% and 17.6%, respectively. V. parahaemolyticus was found in 4.0% of the winter samples, 13.3% of the spring samples, 18.6% of the summer samples and 8% of the autumn samples. During the period of this study, two tdh-positive strains were isolated, while no trh-positive V. parahaemolyticus strain was detected in samples of different seasons.