超高压处理对副溶血性弧菌胞内蛋白质的影响

通过研究超高压处理对水产品源副溶血性弧菌细胞内可溶性蛋白质的影响,进一步探讨超高压处理灭活微生物的机理。从水产品中分离获得的副溶血性弧菌(V.?parahaemolyticus?D)和其经超高压驯化后的副溶血性弧菌(V.?parahaemolyticus?NYD)两株菌经0、100、150、200、250 MPa压力处理后,进行活菌计数,胞内蛋白质十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE)以及细胞内溶物泄漏量测定和胞内蛋白质和总巯基含量测定。结果表明,在250 MPa条件下处理10 min,菌株D和菌株NYD完全失活,培养上清液中核酸物质和蛋白质泄漏量达到最大。随着处理压力的增加,两株菌胞内分离得到的蛋白质的蛋白含量和总巯基含量下降。SDS-PAGE分析结果表明,超高压处理对两株菌分子质量为140~180 k D的胞内蛋白质影响最大。超高压处理可造成副溶血性弧菌细胞内蛋白质变性从而导致细胞死亡。     

酸性电解水对副溶血性弧菌和单核细胞增生李斯特氏菌杀菌效果的比较研究

目的研究不同浓度酸性电解水对副溶血性弧菌和单核细胞增生李斯特氏菌单增李斯特菌的杀菌作用,比较酸性电解水对革兰氏阴性菌和革兰氏阳性菌的杀菌效果。方法采用传统的平板计数法进行细菌计数,扫描电镜观察细菌细胞的形态变化,琼脂糖胶电泳检测细菌DNA变化和BSA法进行细菌蛋白质泄露测定。结果酸性电解水对副溶血性弧菌和单核细胞增生李斯特氏菌均具有很强的杀灭效果,其杀菌效果随着电解水浓度的增加而增强。较高浓度的电解水处理后,副溶血性弧菌几乎全部杀灭,单核细胞增生李斯特氏菌菌落总数降低了1.45 log CFU/mL。扫描电镜实验表明,经酸性电解水处理的副溶血性弧菌坍塌明显,且随着酸性电解水浓度的升高,细胞形态崩解严重,细菌形状模糊。结论相较于单核细胞增生李斯特氏菌,酸性电解水对革兰氏阴性菌副溶血性弧菌有明显的杀菌效果     

Efficacy of High Hydrostatic Pressure Treatment in Reducing Escherichia coli O157 and Listeria monocytogenes in Alfalfa Seeds

ABSTRACT: The application of high hydrostatic pressure (HHP) technology as a seed decontamination technique was evaluated. Alfalfa seeds inoculated with Escherichia coli O157 and Listeria monocytogenes were air-dried and subjected to independent HHP treatments of 275 to 575 MPa for 2 min or at 475 MPa for 2 to 8 min (40°C). There were 1.4-log and 2.0-log reductions in E. coli O157 populations at 575 MPa (2 min) and 475 MPa (8 min), respectively. However, these treatments caused only 0.8-log and 1.1-log reductions in L. monocytogenes counts. Treated seeds took longer to germinate, achieving germination rate of up to 34%, whereas 95% of the control germinated. Results suggest that L. monocytogenes is more resistant to the bactericidal effects of HHP than E. coli O157. Although HHP treatments achieved a greater reduction in E. coli O157, it was at the expense of seed germination. Overall, our results indicate that although HHP treatments reduced the populations of E. coli O157 and L. monocytogenes in alfalfa seeds, they did not completely eliminate these microorganisms.     

Potato Cubes Response to Water Blanching and High Hydrostatic Pressure

High hydrostatic pressure treatment (400 MPa, 15 min, 5–50°C) of potato cubes (2 × 2 × 2 cm) was compared with hot water blanching (100°C, 30–180 set). Inactivation of microorganisms and polyphenoloxidase, texture and density change, leaching of potassium and loss of ascorbic acid were monitored. Reduction of microbial counts was comparable for both methods and polyphenoloxidase was completely inactivated at 20°C, when dilute citric acid solutions (0.5 or 1.0%) were used as “immersion” medium. Hot water blanching and high pressure treatment resulted in samples of similar softness. Leaching of potassium was reduced 20% after pressure treatment on vacuum-packaged samples. Retention of ascorbic acid in pressure-treated samples was temperature dependent ranging from = 90% at 5°C to = 35% at 50°C.     

酸性电解水结合超高压技术对虾仁中副溶血性弧菌风险降低的研究

本研究首次将酸性电解水与超高压技术相结合,用于杀灭虾仁中的副溶血性弧菌,并使用定量微生物风险评估模型对这项杀菌技术的风险降低能力进行了评价。结果表明,这种新型的杀菌技术可显著地降低虾仁中副溶血性弧菌的患病风险,与未经处理的虾仁相比,处理后的虾仁中副溶血性弧菌的最终污染量从1.98 Log CFU/g降低到-2.53 Log CFU/g,平均患病概率从1.28×10~(-1)降低到2.01×10~(-3),每万人中平均患病人数从1280人降低到20人,风险降低率为98%。敏感性分析显示,该技术的处理压力、处理时间及制备电解水时所需的NaCl浓度均与患病概率呈负相关性。此外,本研究还表明该技术可与冷链物流技术相结合,服务于食品工业的生产链,从而最大限度地降低副溶血性弧菌的患病风险,为维护公共卫生提供强有力的技术支持。     

Response of Vibrio parahaemolyticus to ethanol shock

In this study, the growth and survival of Vibrio parahaemolyticus in the presence of 0.0-8.0% ethanol was first examined. V. parahaemolyticus was then exposed to a sub-lethal dose of 5.0% ethanol for 30 and 60 min (ethanol shock). Morphological changes and alterations in cell leakage, thermal tolerance at 47 degrees C, and susceptibility to 8% ethanol and low temperature (4 and -18 degrees C) of V. parahaemolyticus caused by ethanol shock were investigated. In addition, recoveries of the ethanol-shocked cells of V. parahaemolyticus on thiosulfate-citrate-bile salts-sucrose agar (TCBS) and TSA-3.0% NaCl were also compared. The findings revealed that the presence of ethanol in TSB-3.0% NaCl at 6.0-8.0% and 5.0% or less, exerted bactericidal and partial growth inhibition effect, respectively, on V. parahaemolyticus. Recovery of ethanol-shocked cells of V. parahaemolyticus was significantly (P<0.05) less on TCBS than on TSA-3.0% NaCl. A significantly (P<0.05) marked increase of protein and nucleic acid material in the supernatant of cell suspension was found after cells of V. parahaemolyticus were exposed to ethanol shock. Extensive cell disruption, wrinkling and cell-wall pitting, indicative of cell-surface damage were also noted on the ethanol-shocked cells. Ethanol-shocked cells of V. parahaemolyticus exhibited a similar yet higher susceptibility at 4 and -18 degrees C compared with the control cells. Moreover, there was a marked increase in the thermal tolerance and resistance to 8.0% ethanol with cells of V. parahaemolyticus after ethanol shock. Finally, the duration of ethanol shock testing did not affect the extent of increased thermal tolerance. While cells of V. parahaemolyticus subjected to ethanol shock for 60 min showed an increase in their resistance to 8.0% ethanol, they also showed an increase in susceptibility at -18 degrees C, than those ethanol shocked for 30 min.     

多重荧光定量PCR法检测海产品中副溶血性弧菌、沙门菌和单增李斯特菌

目的建立检测海产品中副溶血性弧菌、沙门菌和单增李斯特菌的多重荧光定量PCR体系。方法针对副溶血性弧菌tlh基因,沙门菌Ompc基因和单增李斯特菌hly基因设计引物和Taq Man探针,建立多重荧光定量PCR体系,进行特异性与敏感性研究;利用该体系检测海产品中的副溶血性弧菌、沙门菌和单增李斯特菌。结果副溶血性弧菌、沙门菌和单增李斯特菌可得到特异性扩增,而共存于海产品中的其他细菌均未见扩增曲线。敏感性试验显示,该体系对副溶血性弧菌、沙门菌和单增李斯特菌的最低检测限分别为72、40、80 cfu/ml。对舟山采集的150份样品进行检测,检出32份副溶血性弧菌、11份沙门菌、5份单增李斯特菌,与国标法检测结果一致。结论本研究建立的基于Taq Man探针的多重荧光定量PCR检测方法可以特异、灵敏、简单快速地实现对海产品中副溶血性弧菌、沙门菌和单增李斯特菌的检测。     

Reuterin and High Hydrostatic Pressure Treatments on the Inactivation of Listeria monocytogenes and Effect on the Characteristics of Cold-Smoked Salmon

The effect of reuterin and high hydrostatic pressure (HHP) processing at 450 MPa for 5 min on the inactivation of Listeria monocytogenes and the characteristics of cold-smoked salmon during 35 days at 4 and 10 °C were investigated. The growth rate of the pathogen was reduced by reuterin addition and a synergistic antimicrobial effect against L. monocytogenes was recorded when the biopreservative was applied in combination with HHP at 450 MPa for 5 min. This combined treatment prevented the pathogen recovery observed with individual treatments and delayed the spoilage of smoked salmon maintaining total viable counts under 3.5 log units during 35 days of storage at 4 °C. All treatments assayed induced changes in lightness (L*) and redness (a*), resulting in a brighter appearance of smoked salmon, whereas no modifications were recorded in shear strength values immediately after treatments. Moreover, reuterin and HHP treatments, individually or in combination, avoided the formation of biogenic amines during the 35 days of storage at 4 and 10 °C. The addition of reuterin in combination with HHP at 450 MPa for 5 min might be applied as a hurdle technology to improve the safety and extend the shelf life of lightly preserved seafood products, such as cold-smoked salmon.     

Response of heat-shocked Vibrio parahaemolyticus to subsequent physical and chemical stresses

Vibrio parahaemolyticus foodborne strains 405, 556, and 690 and a V. parahaemolyticus chopping board isolate were heat shocked at 42 degrees C for 15, 30, or 45 min. Heat shock, regardless of heating periods tested, caused an increased demand for NaCl during recovery from heat injury. Further study with strain 690 and the chopping board isolate also revealed that heat shock generally increased the survival of the test organism during subsequent exposure to 47 degrees C, 20 ppm H202, and 8% ethanol and reduced the tolerance of the test organism to low temperatures (5 and -18 degrees C). The extent of the heat shock response of V. parahaemolyticus varied with strain and the duration of treatment. Furthermore, heat shock treatments in the present study caused the leakage of nucleic acids from V. parahaemolyticus cells. This effect was most pronounced with cells heat shocked at 42 degrees C for 45 min.     

High Hydrostatic Pressure Processing of Cheese

Abstract: High hydrostatic pressure (HHP) is a cutting‐edge processing technology attracting research and industrial interest in the food sector due to its potential to produce microbiologically safe products, modify the functional properties of proteins and polysaccharides, and alter biochemical reactions without significantly affecting the nutritional and sensory properties of food. Currently, there are only a limited number of pressure‐treated cheese products available in the market. Nevertheless, results from numerous research studies on various cheese varieties seem promising, especially since HHP technology is today more cost‐effective than in the past. Considering the progress made in the application of HHP on cheese during the past 15 years, this paper reviews the direct application of HHP treatments to cheese and the effects it has on its microbiology and ripening process, as well as on quality parameters such as physicochemical, rheological, and sensory properties. Detailed information of published studies is presented with the aim of providing a clear picture of the use of this technology on cheese processing. Areas of research in need of more attention are also identified.     

低温条件下即食虾中单增李斯特菌与副溶血性弧菌共存分子预测模型的建立

本研究旨在应用实时荧光定量聚合酶链式反应（quantitative real-time polymerase chain reaction,qPCR）技术描述即食虾中的单增李斯特菌与副溶血性弧菌的生长行为,构建混菌模式下的分子预测模型。将单增李斯特菌与副溶血性弧菌等量（（5.0±0.5）（lg（CFU/g）））混合接种于即食虾中,置于低温环境（4 ℃和10 ℃）下培养,并利用qPCR定量检测单增李斯特菌与副溶血性弧菌数量的动态变化。运用生长模型（修正Gompertz、Logistic、Baranyi）和失活模型（Log-linear、Weibull）分别拟合两株菌的生长和失活趋势。结果表明：低温条件下,修正Gompertz、Logistic和Baranyi模型均可成功拟合单增李斯特菌的生长曲线,其决定系数（R2）均大于0.98。对于副溶血性弧菌,在4 ℃条件下,Log-linear和Weibull模型能够清晰地描述其失活情况,R2分别为0.950和0.945;而10 ℃条件下,2 个失活模型均难以描述其行为变化,R2仅为0.784和0.775。本研究运用分子生物学技术描述即食虾中两种致病菌混合培养的菌量变化,探究混菌模式下微生物的生长失活情况,为分子预测模型的进一步研究提供了新的思路。     

Gelation of Surimi by High Hydrostatic Pressure

The effects of high hydrostatic pressure (HHP) on gel strength of Pacific whiting and Alaska pollock surimi were determined by torsion. Pacific whiting gels were made with and without 1% beef plasma protein (BPP) as protease inhibitor. HHP treated whiting (1% BPP added) and pollock gels showed greatly increased strain values at all pressure/temperature combinations compared with heat-set controls. Stress values for the same samples were variable depending on treatment and species. A three-fold increase in strain and stress was found for HHP treated whiting gels made without inhibitor.     

High Hydrostatic Pressure Pasteurization of Red Wine

ABSTRACT:  Pasteurization of low-alcohol wine using a high hydrostatic pressure (HHP) process was studied. A total of 10 mL grape wine sealed in a nylon/LLDPE bag was placed inside the HHP chamber. The pressure applied to the treatment chamber was maintained at 1000 to 3500 atm for 0 to 30 min. The effects of HHP treatments on the physiochemical properties (alcohol, pH, acidity, total sugar) and microbes (aerobic bacteria, yeast, and lactic acid bacteria) were examined. The HHP treatments had little impact on the physiochemical properties. The pasteurization effect of the HHP treatments increased with treatment pressure and time. A total of 2 different stages in the microbial inactivation were noticed when the 1st-order reaction model was used to fit the inactivation data. The inactivation rate was higher in the initial stages than in the later stages, suggesting that might be 2 different groups of the microorganisms, a more HHP-susceptible group and a less HHP-susceptible group.     

Evaluation of High Pressure Processing as an Additional Hurdle to Control Listeria monocytogenes and Salmonella enterica in Low-Acid Fermented Sausages

Low‐acid fermented sausages (fuet and chorizo) were manufactured to evaluate the combined effect of high pressure processing (HPP) and ripening on foodborne pathogens. Raw sausages inoculated with a three‐strain cocktail of Salmonella ser. Derby, London, and Schwarzengrund, and a three‐strain cocktail of L. monocytogenes ser. 1/2 c and 4b were pressurized at 300 MPa for 10 min at 17 °C. Afterwards, sausages were ripened at 12 °C and 80% RH for 27 d. Salmonella counts decreased in all studied sausages during ripening. However, the application of HPP as an additional hurdle to the ripening process produced a greater decrease in the Salmonella population, showing lower counts (3 MPN/g) in ripened sausages. By contrast, lower values of L. monocytogenes counts were obtained in non‐treated (NT) than in pressurized sausages due to the delay of pH drop caused by HPP inactivation of endogenous lactic acid bacteria. After pressurization of raw sausages at 300 MPa, a discoloration of sausages was observed, coinciding with an increase in L* values.     

High Hydrostatic Pressure Effects on Vegetable Structure

Cryo-fracture scanning electron microscopy revealed the changes that cauliflower and spinach leaves undergo after treatment with high hydrostatic pressure. High pressure changed cell permeability and enabled movement of water and metabolites from inside to outside of the cell. The ultrastructure showed that parenchyma organization disappeared in spinach leaf and cavity formation occurred after treatment. However, cauliflower exhibited a firm structure with a soaked appearance and would be more suitable than spinach for high hydrostatic pressure treatment.     

High Hydrostatic Pressure Treatments for Beer Stabilization

ABSTRACT: Crude cloudy beers from a small-scale brewery were treated with high hydrostatic pressure (HHP-600 MPa, 5 min) or heat pasteurized (60 °C, 10 min). The treatments did not affect pH, ethanol, extract, bitterness, or phenolics in comparison with untreated beers; HHP beers retained permanent haze, similar to untreated samples. Heat pasteurized beer resulted in a more rapid increase of the a* (red) and a slower decrease of the b* (yellow) color parameters than in HHP samples. The microbiological stability of HHP beers was comparable with heat-treated beers, and the development of yeast and lactic acid bacteria was inhibited for 49 d of storage.     

High Hydrostatic Pressure Processing of Fruit and Vegetable Products

High hydrostatic pressure (HHP) as a minimal thermal technology is a valuable tool for microbiologically safe and shelf-stable fruit and vegetable production. Microorganisms and deteriorative enzymes can be inhibited or inactivated depending on the amount of pressure and time applied to the product. The resistance of microorganisms and enzymes to pressure in fruit and vegetable products also is dependent on both the type and the amount of enzymes or microorganisms as well as food composition. While on one hand, microorganisms (other than spores) can be inactivated at mild pressures (< 300 MPa), on the other, enzymes can be very resistant to pressure and their resistance may increase when isolated forms are pressurized. Nevertheless, microbiologically safe fruit and vegetable products can be obtained without change in flavor if temperature is not increased beyond pasteurization temperatures. The remaining enzyme activity in HHP processed fruit and vegetable products can be delayed if a combination of obstacles, such as refrigeration temperatures, low pH, and antibrowning agents, are used to increase the shelf life of these types of products. Therefore, HHP is a promising minimal thermal technology that can be used to deliver more variety of less processed fruit and vegetable products than consumers are demanding today.     

High Salinity Relaying to Reduce Vibrio parahaemolyticus and Vibrio vulnificus in Chesapeake Bay Oysters (Crassostrea virginica)

Cases of Vibrio infections in the United States have tripled from 1996 to 2009 and these infections are most often associated with the consumption of seafood, particularly oysters (Crassostrea virginica). Information is needed on how to reduce numbers of Vibrio parahaemolyticus and Vibrio vulnificus in bi‐valve molluscan shellfish (for example, oysters). The purpose of this study was to evaluate the effectiveness of high salinity relaying or treatment in recirculating aquaculture systems (RASs) as methods to reduce the abundance of V. parahaemolyticus and V. vulnificus in oysters. For relaying field trials, oysters were collected from approved harvest waters, temperature abused outside under a tarp for 4 h, and then transferred to high (29 to 33 ppt.) and moderate (12 to 19 ppt.) salinities. For RAS treatment trial, oysters were transferred to 32 to 34 ppt. salinity at 15 °C. After 7, 14, 21, and in some instances 28 d, oysters were collected and analyzed for V. parahaemolyticus and V. vulnificus levels using multiplex real‐time PCR. Initial levels of V. parahaemolyticus and V. vulnificus ranged from 3.70 to 5.64 log₁₀ MPN/g, and were reduced by 2 to 5 logs after 21 to 28 d in high salinity water (29 to 34 ppt.). Oyster mortalities averaged 4% or less, and did not exceed 7%. Relaying of oysters to high salinity field sites or transfer to high salinity RAS tanks was more effective in reducing V. vulnificus compared with V. parahaemolyticus. These results suggest that high salinity relaying of oysters is more effective in reducing V. vulnificus than V. parahaemolyticus in the oyster species used in this study.