Recent advances in understanding the fitness and survival mechanisms of Vibrio parahaemolyticus.

The presence of Vibrio parahaemolyticus (Vp) in different production stages of seafood has generated negative impacts on both public health and the sustainability of the industry. To further better investigate the fitness of Vp at the phenotypical level, a great number of studies have been conducted in recent years using plate counting methods. In the meantime, with the increasing accessibility of the next generation sequencing and the advances in analytical chemistry techniques, omics-oriented biotechnologies have further advanced our knowledge in the survival and virulence mechanisms of Vp at various molecular levels. These observations provide insights to guide the development of novel prevention and control strategies and benefit the monitoring and mitigation of food safety risks associated with Vp contamination. To timely capture these recent advances, this review firstly summarizes the most recent phenotypical level studies and provide insights about the survival of Vp under important in vitro stresses and on aquatic products. After that, molecular survival mechanisms of Vp at transcriptomic and proteomic levels are summarized and discussed. Looking forward, other newer omics-biotechnology such as metabolomics and secretomics show great potential to be used for confirming the cellular responses of Vp. Powerful data mining tools from the field of machine learning and artificial intelligence, that can better utilize the omics data and solve complex problems in the processing, analysis, and interpretation of omics data, will further improve our mechanistic understanding of Vp.

Pilot Scale Assessment of High-Pressure Processing (HPP) to Enhance Microbiological Quality and Shelf Life of Fresh Ready-to-Eat (RTE) Blue Crab Meat.

Blue crab (Callinectes sapidus) is a highly valuable wild fishery species of crab native to the waters of the western Atlantic Ocean and the Gulf of Mexico. The annual commercial production of live blue crabs is approximately 50,000 metric tons with a dockside value of USD 200 million. Presently the US blue crab processing industry sells crab meat in three basic forms: fresh crab meat, pasteurized crab meat, and frozen crab meat. By far "Fresh" is the most desirable form of crab meat. However, fresh crab meat has a limited shelf life. This study evaluated the effects of high-pressure processing (HPP) on enhancing the microbiological quality and shelf life of blue crab meat. Live blue crabs were pressure-cooked in a retort (≥115 °C for 4-6 min). The crab meat was handpicked, packed in plastic containers with seals, subjected to HPP treatment, and stored at 4 °C. Container integrity and water leakage issues were examined by observation in addition to weight comparison before and after HPP treatment; the shelf life of crab meat with and without HPP treatments was examined via microbiological tests and sensory evaluations. Results show that polypropylene containers sealed with 10K OTR (oxygen transmission rate) film could withstand high pressure without water leakage issues; HPP treatment at 600 MPa for 3 min could extend the shelf life of fresh, cooked, and handpicked crab meat from 6 days to 18 days based on the strictest APC (aerobic plate account) limit (APC ≤ 100,000 CFU/g). The sensory quality of the HPP-treated crab meat was well accepted throughout the 3-week storage period. The results support the use of HPP as an effective non-thermal processing technology to enhance the microbiological quality and extend the shelf life of fresh RTE blue crab meat.

Controlled Recirculating Wet Storage Purging V. parahaemolyticus in Oysters.

This work explored the effects of salinity and temperature on the efficacy of purging V. parahaemolyticus from eastern oysters (Crassostrea virginica). Oysters were inoculated with a 5-strain cocktail of V. parahaemolyticus to levels of 104 to 105 MPN (most probable number)/g and depurated in a controlled re-circulating wet-storage system with artificial seawater (ASW). Both salinity and temperature remarkably affected the efficacy for the depuration of V. parahaemolyticus from oysters during wet-storage. The wet-storage process at salinity 20 ppt at 7.5 °C or 10 °C could achieve a larger than 3 log (MPN/g) reduction of Vibrio at Day 7, which meets the FDA's requirement as a post-harvest process for V. parahaemolyticus control. At the conditions of 10 °C and 20 ppt, a pre-chilled system could achieve a 3.54 log (MPN/g) reduction of Vibrio in oysters on Day 7. There was no significant difference in the shelf life between inoculated and untreated oysters before the depuration, with a same survival rate (stored in a 4 °C cooler for 15 days) of 93%.

Seafood and health: What you need to know?

Seafood, including fish and shellfish, provides an ideal package of nutrients and is an important part of a healthy diet. Strong evidence has shown that eating fish and other seafoods improve brain, eye, and heart health. The new 2020-2025 Dietary Guidelines for Americans (DGA) recommend that Americans of all ages should eat more seafood-at least twice a week-particularly pregnant women and young children. However, less than one in five Americans heed that advice. About one-third of Americans eat seafood once a week, while nearly half eat fish only occasionally or not at all. This calls for a drastic shift in the American diet to vary protein sources and include more seafood products in order to receive the most health benefits. This chapter covers (1) seafood nutrition and health benefits, (2) seafood's protective effects against mercury toxicity, (3) selenium health benefit values (HBVs), and (4) challenges and opportunities for seafood production, demand and sustainability. This chapter aims to convey recent advances in science-based information to increase public awareness of seafood safety, nutrition and health benefits of seafood as part of a healthy diet, and to advocate healthy eating with smart food choices by promoting two servings of seafood per week. This will support the healthy eating patterns and promotes a minimum two to three servings of seafood recommended by the current DGA.

Efficacy of Vibrio parahaemolyticus depuration in oysters (Crassostrea gigas).

This study investigated the influences of seawater to oyster ratio on depuration for decontaminating V. parahaemolyticus in raw oysters with a goal of identifying the proper ratio of oyster to seawater capable of improving the efficacy of the depuration process. The water to oyster ratios tested in this study ranged from 1.0 to 2.5 L of artificial seawater (ASW) per oyster (40 oysters in 40, 60, 80 and 100 L ASW). The depuration efficacy for purging V. parahaemolyticus from oysters was highest when we applied a 2:1, followed by 1.5:1, 2.5:1, and 1:1 L of ASW/oyster. Further studies of depuration with 2:1 L of ASW/oyster found that the concentration of V. parahaemolyticus in oysters decreased in a nonlinear manner. The depuration curve was fitted to a one phase decay model with a coefficient of determination (R2) of 0.933. The time for a 3 log reduction was 1.75 days with a 95% confidence interval from 1.65 to 1.85 days, which meets the FDA's requirement of larger than a 3.0 log (MPN/g) reduction as a post-harvest process for V. parahaemolyticus control. After 4 days levels in all trials were <100 MPN/g meeting performance standards established by Japan and Canada. Furthermore, the time for a 3.52 log reduction was 3.17 days with a 95% confidence interval from 2.92 to 3.54 days but it took 5 days to reduce levels to <30 MPN/g, which satisfies FDA's requirement as a post-harvest control process (>3.52 log MPN/g reduction) for the purpose of making safety added labeling claims for V. parahaemolyticus.

Behavior of Salmonella and Listeria monocytogenes in Raw Yellowfin Tuna during Cold Storage.

Behavior of Salmonella and Listeria monocytogenes in raw yellowfin tuna during refrigeration and frozen storage were studied. Growth of Salmonella was inhibited in tuna during refrigerated storage, while L. monocytogenes was able to multiply significantly during refrigerated storage. Populations of Salmonella in tuna were reduced by 1 to 2 log after 12 days of storage at 5-7 °C, regardless levels of contamination. However, populations of L. monocytogenes Scott A, M0507, and SFL0404 in inoculated tuna (104-105 CFU/g) increased by 3.31, 3.56, and 3.98 log CFU/g, respectively, after 12 days of storage at 5-7 °C. Similar increases of L. monocytogenes cells were observed in tuna meat with a lower inoculation level (10²-10³ CFU/g). Populations of Salmonella and L. monocytogenes declined gradually in tuna samples over 84 days (12 weeks) of frozen storage at -18 °C with Salmonella Newport 6962 being decreased to undetectable level (<10 CFU/g) from an initial level of 10³ log CFU/g after 42 days of frozen storage. These results demonstrate that tuna meat intended for raw consumption must be handled properly from farm to table to reduce the risks of foodborne illness caused by Salmonella and L. monocytogenes.

[The study on the fluorescence spectrometry for a novel photosensitizer of chlorine e6-C15-monomethyl ester and its application in biosamples analysis].

This paper studied the fluorescence spectral characteristic of chlorin e6-C15-monomethyl ester in different solvents to develop a fluorescence spectrometry for determining the concentration of chlorin e6-C15-monomethyl ester in plasma. By comparing the fluorescence spectral characteristics of chlorin e6-C15-monomethyl ester in six different solvents including methanol, ethanol, acetone, acetonitrile, phosphate buffered saline and water, the influence of different solvents on the fluorescence spectral characteristic of chlorin e6-C15-monomethyl ester has been examined. The results indicated that methanol and acetonitrile are the ideal solvent system, and the effect of phosphate buffered saline is better than water. The effect of organic solvent content and solution pH value on emission wavelength and fluorescence intensity was further evaluated. It was found that the fluorescence intensity of chlorin e6-C15-monomethyl ester was strong and stable in pH 7.2 phosphate buffered saline -acetonitrile (3 : 7) solution and can be detected at the excitation wavelength of 498.00 nm and the emission wavelength of 664.05 nm. Based on this, we developed the fluorescence spectrometry for determining chlorin e6-C15-monomethyl ester. It is specific and sensitive with good linearity over the range of 0.5-50 microg x mL(-1). The intra-batch and inter-batch precisions (RSD) were less than 10%, the extraction recoveries were all over 90%. The established method in this paper was simple, fast, effective and could be successfully applied to study the pharmacokinetics of chlorine e6-C15-monomethyl ester in SD rats.

Degradable copolymer based on amphiphilic N-octyl-N-quatenary chitosan and low-molecular weight polyethylenimine for gene delivery.

BACKGROUND: Chitosan shows particularly high biocompatibility and fairly low cytotoxicity. However, chitosan is insoluble at physiological pH. Moreover, it lacks charge, so shows poor transfection. In order to develop a new type of gene vector with high transfection efficiency and low cytotoxicity, amphiphilic chitosan was synthesized and linked with low-molecular weight polyethylenimine (PEI). METHODS: We first synthesized amphiphilic chitosan - N-octyl-N-quatenary chitosan (OTMCS), then prepared degradable PEI derivates by cross-linking low-molecular weight PEI with amphiphilic chitosan to produce a new polymeric gene vector (OTMCS-PEI). The new gene vector was characterized by various physicochemical methods. We also determined its cytotoxicity and gene transfecton efficiency in vitro and in vivo. RESULTS: The vector showed controlled degradation. It was very stable and showed excellent buffering capacity. The particle sizes of the OTMCS-PEI/DNA complexes were around 150-200 nm with proper zeta potentials from 10 mV to 30 mV. The polymer could protect plasmid DNA from being digested by DNase I at a concentration of 2.25 U DNase I/µg DNA. Furthermore, they were resistant to dissociation induced by 50% fetal bovine serum and 1100 µg/mL sodium heparin. OTMCS-PEI revealed lower cytotoxicity, even at higher doses. Compared with PEI 25 KDa, the OTMCS-PEI/DNA complexes also showed higher transfection efficiency in vitro and in vivo. CONCLUSION: OTMCS-PEI was a potential candidate as a safe and efficient gene vector for gene therapy.

[Photodynamic effect of two kinds of phycobiliproteins on human liver cancer cell line SMMC-7721 in vitro].

We studied the effect of photodynamic therapy with phycobiliproteins on human liver cancer cells in vitro. With 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT assay), we used two phycobiliproteins, R-phycoerythrin (R-PE) and C-phycocyanin (C-PC) prepared from Porphyra yezoensis, to determine the killing rates and apoptosis rates of human liver cancer cells (SMMC-7721) mediated by laser. When the concentration of R-PE was 120 mg/L, the survival rate of human liver cancer cells was 27% after treated by Argon laser with 100 J/cm2 doses, while the survival rate in the control group (without adding R-PE) was 65%. When the C-PC concentration was 120 mg/L, the survival cell rate was 47% after treated by He-Ne laser with 35 J/cm2 dose, while the survival rate in the control group (without adding C-PC) was 70%. After handled only with these two kinds of phycobiliproteins for 72 h, the growth of cancer cells presented significant inhibition. The maximal inhibition rates reached up to 31% with R-PE (120 mg/L concentration) and 27% with C-PC (250 mg/L concentration) respectively. After irradiated by laser for 8 h, the maximal cell apoptosis rates were 31.54% with R-PE and 32.54% with C-PC, respectively. It indicated that R-PE and C-PC extracted from Porphyra yezoensis could develop to new photosensitizers for cancer photodynamic therapy.

Effect of temperature on uptake and survival of Vibrio parahaemolyticus in oysters (Crassostrea plicatula).

This study investigated accumulation of Vibrio parahaemolyticus in Zhe oyster (Crassostrea plicatula) from culture water and effectiveness of frozen and chilled storage on reducing V. parahaemolyticus in oysters. Freshly harvested oysters were placed in artificial seawater containing V. parahaemolyticus (10(4)CFU/mL) at 16, 20, 26, and 32 degrees C for 96 h. Contaminated oysters were stored at chilled temperatures (0, 5, and 15 degrees C) and frozen at -18 and -30 degrees C and changes of V. parahaemolyticus populations in oysters were determined using the most probable number (MPN) method. Accumulations of V. parahaemolyticus in C. plicatula reached the peaks at 6.66 (32 degrees C), 5.72 (26 degrees C), 5.04 (20 degrees C), 4.72 (16 degrees C) log MPN/g after 32 h in contaminated artificial seawater. Holding contaminated Zhe oysters at 5 and 0 degrees C reduced V. parahaemolyticus populations in both shell stock and shucked oysters. Populations of V. parahaemolyticus in shell stock and shucked oysters declined by 1.42 and 2.0 log MPN/g, respectively, after 96 h of storage at 5 degrees C and by 2.11 and 2.38 log MPN/g, respectively, after 96 h of storage at 0 degrees C. However, populations of V. parahaemolyticus increased by 2.44 log MPN/g in shell stock oysters and by 1.64 og MPN/g in shucked oysters when stored at 15 degrees C for 60 h. Frozen storage was effective in inactivating V. parahaemolyticus. Populations of V. parahaemolyticus in shell stock and shucked oysters decreased from 5.46log MPN/g to 1.66 and 0.38 log MPN/g, respectively, after 75 days of storage at -30 degrees C. No V. parahaemolyticus cells were detected (<3 log MPN/g) in the shucked oysters after 60 days of storage at -18 degrees C. These results demonstrated that accumulation of V. parahaemolyticus in cultured C. plicatula increases as water temperature increases. Harvested C. plicatula should be stored at 5 degrees C or lower to control the hazard of V. parahaemolyticus.

Effects of flash freezing, followed by frozen storage, on reducing Vibrio parahaemolyticus in Pacific raw oysters (Crassostrea gigas).

This study investigated the effects of flash freezing, followed by frozen storage, on reducing Vibrio parahaemolyticus in Pacific raw oysters. Raw Pacific oysters were inoculated with a five-strain cocktail of V. parahaemolyticus at a total level of approximately 3.5 x 10(5) most probable number (MPN) per gram. Inoculated oysters were subjected to an ultralow flash-freezing process (-95.5 degrees C for 12 min) and stored at -10, -20, and -30 degrees C for 6 months. Populations of V. parahaemolyticus in the oysters declined slightly by 0.22 log MPN/g after the freezing process. Subsequent storage of frozen oysters at - 10, -20, and -30 degrees C resulted in considerable reductions of V. parahaemolyticus in the oysters. Storing oysters at -10 degrees C was more effective in inactivating V. parahaemolyticus than was storage at -20 or -30 degrees C. Populations of V. parahaemolyticus in the oysters declined by 2.45, 1.71, and 1.45 log MPN/g after 1 month of storage at -10, -20, and -30 degrees C, respectively, and continued to decline during the storage. The levels of V. parahaemolyticus in oysters were reduced by 4.55, 4.13, and 2.53 log MPN/g after 6 months of storage at -10, -20, and -30 degrees C, respectively. Three process validations, each separated by 1 week and conducted according to the National Shellfish Sanitation Program's postharvest processing validation-verification interim guidance for Vibrio vulnificus and Vibrio parahaemolyticus, confirmed that a process of flash freezing, followed by storage at -21 +/- 2 degrees C for 5 months, was capable of achieving greater than 3.52-log (MPN/g) reductions of V. parahaemolyticus in half-shell Pacific oysters.

Vibrio parahaemolyticus: a concern of seafood safety.

Vibrio parahaemolyticus is a human pathogen that is widely distributed in the marine environments. This organism is frequently isolated from a variety of raw seafoods, particularly shellfish. Consumption of raw or undercooked seafood contaminated with V. parahaemolyticus may lead to development of acute gastroenteritis characterized by diarrhea, headache, vomiting, nausea, and abdominal cramps. This pathogen is a common cause of foodborne illnesses in many Asian countries, including China, Japan and Taiwan, and is recognized as the leading cause of human gastroenteritis associated with seafood consumption in the United States. This review gives an overview of V. parahaemolyticus food poisoning and provides information on recent development in methods for detecting V. parahaemolyticus and strategies for reducing risk of V. parahaemolyticus infections associated with seafood consumption.

Bactericidal effects of wine on Vibrio parahaemolyticus in oysters.

The bactericidal effects of wines on Vibrio parahaemolyticus in oysters were studied to evaluate potential inactivation of V. parahaemolyticus in contaminated oysters by wine consumption. Shucked whole oyster and oyster meat homogenate were inoculated with V. parahaemolyticus and mixed with red or white wine. Survivals of V. parahaemolyticus in inoculated oysters were determined at 7 and 25 degrees C. Populations of V. parahaemolyticus in inoculated whole oysters (5.52 log most probable number [MPN] per g) decreased slightly to 4.90 log MPN/g (a 0.62-log reduction) after 24 h at 7 degrees C but increased to 7.37 log MPN/g over the same period at 25 degrees C. However, the populations in wine-treated whole oysters decreased by >1.7 and >1.9 log MPN/g after 24 h at 7 and 25 degrees C, respectively. Both red and white wines were more effective in inactivating V. parahaemolyticus in oyster meat homogenate than in whole oyster. Populations of V. parahaemolyticus in oyster meat homogenate (7.8 x 10(3) MPN/g) decreased rapidly to nondetectable levels (< 3 MPN/g) after 30 min of mixing with wine at 25 degrees C (a 3.89-log MPN/g reduction). These results suggest that chewing oysters before swallowing when eating raw oysters may result in greater inactivation of V. parahaemolyticus if wine is consumed. More studies are needed to determine the bactericidal effects of wine on V. parahaemolyticus in the complicated stomach environment.

Efficiency of electrolyzed oxidizing water on reducing Listeria monocytogenes contamination on seafood processing gloves.

Food processing gloves are typically used to prevent cross-contamination during food preparation. However, gloves can be contaminated with microorganisms and become a source of contamination. This study investigated the survival of Listeria monocytogenes on gloves and determined the efficacy of electrolyzed oxidizing (EO) water for reducing L. monocytogenes contamination on seafood processing gloves. Three types of reusable gloves (natural rubber latex, natural latex, and nitrile) and two types of disposable gloves (latex and nitrile) were cut into small pieces (4 x 4 cm(2)) and inoculated with 5-strain L. monocytogenes cocktail (5.1 x 10(7) CFU/cm(2)) with and without shrimp meat residue attached to surfaces. L. monocytogenes did not survive well on clean reusable gloves and its populations decreased rapidly to non-detectable levels within 30 min at room temperature. However, high levels of Listeria cells were recovered from clean disposable gloves after 30 min of inoculation. Presence of shrimp meat residue on gloves enhanced the survival of L. monocytogenes. Cells of L. monocytogenes were detected on both reusable and disposal gloves even after 2 h at room temperature. Soaking inoculated gloves in EO water at room temperature for 5 min completely eliminated L. monocytogenes on clean gloves (>4.46 log CFU/cm(2) reductions) and significantly (p<0.05) reduced the contamination on soil-containing gloves when compared with tap water treatment. EO water could be used as a sanitizer to reduce L. monocytogenes contamination on gloves and reduce the possibility of transferring L. monocytogenes from gloves to RTE seafoods.

Effects of electrolyzed oxidizing water on reducing Listeria monocytogenes contamination on seafood processing surfaces.

The effects of electrolyzed oxidizing (EO) water on reducing Listeria monocytogenes contamination on seafood processing surfaces were studied. Chips (5 x 5 cm(2)) of stainless steel sheet (SS), ceramic tile (CT), and floor tile (FT) with and without crabmeat residue on the surface were inoculated with L. monocytogenes and soaked in tap or EO water for 5 min. Viable cells of L. monocytogenes were detected on all chip surfaces with or without crabmeat residue after being held at room temperature for 1 h. Soaking contaminated chips in tap water resulted in small-degree reductions of the organism (0.40-0.66 log cfu/chip on clean surfaces and 0.78-1.33 log cfu/chip on dirty surfaces). Treatments of EO water significantly (p<0.05) reduced L. monocytogenes on clean surfaces (3.73 log on SS, 4.24 log on CT, and 5.12 log on FT). Presence of crabmeat residue on chip surfaces reduced the effectiveness of EO water on inactivating Listeria cells. However, treatments of EO water also resulted in significant reductions of L. monocytogenes on dirty surfaces (2.33 log on SS and CT and 1.52 log on FT) when compared with tap water treatments. The antimicrobial activity of EO water was positively correlated with its chlorine content. High oxidation-reduction potential (ORP) of EO water also contributed significantly to its antimicrobial activity against L. monocytogenes. EO water was more effective than chlorine water on inactivating L. monocytogenes on surfaces and could be used as a chlorine alternative for sanitation purpose. Application of EO water following a thorough cleaning process could greatly reduce L. monocytogenes contamination in seafood processing environments.