Effect of simulated gastric fluid and bile on survival of Vibrio vulnificus and Vibrio vulnificus phage

Bacteria and phages may be exposed to acid conditions in the stomach and to bile in the intestine. Survival of three strains of Vibrio vulnificus and three strains of its phages was examined at 37 degrees C after exposure to simulated gastric fluid at pH 3 to 4 or to 0, 1, and 2% bile in broth or buffer. Mean D-values (decimal reduction times) at pH 4 and 3 were 3.3 and 1.3 min for V. vulnificus and 97.8 and 0.7 min for its phages. No V. vulnificus survivors were found at pH 2.0. There were few survival differences among strains of V. vulnificus or its phages. Numbers of V. vulnificus increased 1 log in tryptic soy broth containing 1 or 2% bile after 3 h. Numbers of V. vulnificus and its phages remained constant in phosphate-buffered saline regardless of bile concentrations up to 3 h. Those V. vulnificus bacteria and phages that survive stomach acidity may proliferate in the small intestine, since they are resistant to bile.     

Resistance of cold- and starvation-stressed Vibrio vulnificus to heat and freeze-thaw exposure

The effects of cold storage and starvation on the subsequent heat resistance and freeze-thaw resistance of Vibrio vulnificus were studied. Three strains of V. vulnificus were evaluated. Cold stress had no effect on freeze-thaw resistance (P > 0.05). Starvation enhanced freeze-thaw resistance for one strain compared to controls (P < 0.05). V. vulnificus was not heat resistant; control populations were inactivated within 12 min at 47 degrees C. Starvation increased heat tolerance for one strain, but differences were small from a processing perspective (P < 0.05). Cold stress had no effect on heat resistance (P > 0.05). Cold adaptation (holding 4 h at 15 degrees C) enhanced cold temperature (5 degrees C) tolerance. This information will be helpful in the development of methods to minimize V. vulnificus risk.     

Conditions for a 5-log reduction of Vibrio vulnificus in oysters through high hydrostatic pressure treatment

Vibrio vulnificus is frequently associated with oysters, and since oysters are typically consumed raw on a half shell, they can pose a threat to public health due to ingestion of this pathogenic marine microorganism. Oysters should be processed to reduce the number of this pathogen. High pressure processing is gaining more and more acceptance among oyster processors due to its ability to shuck oysters while keeping the fresh-like characteristics of oysters. Nine strains of V. vulnificus were tested for their sensitivities to high pressure. The most pressure-resistant strain of V. vulnificus, MLT 403, was selected and used in the subsequent experiments to represent a worst case scenario for evaluation of the processing parameters for inactivation of V. vulnificus in oysters. To evaluate the effect of temperature on pressure inactivation of V. vulnificus, oyster meats were inoculated with V. vulnificus MLT 403 and incubated at room temperature for 24 h. Oyster meats were then blended and treated at 150 MPa for 4 min, and 200 MPa for 1 min. Pressure treatments were carried out at -2, 1, 5, 10, 20, 30, 40, and 45 degrees C. Cold temperatures (<20 degrees C) and slightly elevated temperatures (>30 degrees C) substantially increased pressure inactivation of V. vulnificus. For example, a 4-min treatment of 150 MPa at -2 and 40 degrees C reduced the counts of V. vulnificus by 4.7 and 2.8 log, respectively, while at 20 degrees C the same treatment only reduced counts by 0.5 log. Temperatures of -2 and 1 degrees C were used to determine the effect of pressure level, temperature, and treatment time on the inactivation of V. vulnificus infected to live oysters through feeding. To achieve a >5-log reduction in the counts of V. vulnificus in a relatively short treatment time (or=250 MPa at -2 or 1 degrees C.     

低温贮藏条件下创伤弧菌和副溶血性弧菌失活模型的建立

为掌握对虾中创伤弧菌和副溶血性弧菌在5℃和一18~C低温贮藏条件下失活动力学特征,分别采用线性模型、Weibull模型、Logistic模型对创伤弧菌和副溶血性弧菌的失活曲线进行拟合。研究结果表明,在5~C条件下,创伤弧茵VvHB09的耐冷力较强;．18℃条件下,副溶血性弧菌ATCC17802和创伤弧菌VvSH09的耐冷力较强。线性模型比weibull模型、Logistic模型更适合拟合创伤弧菌和副溶血性弧菌的失活特征。     

Recovery and detection of Vibrio vulnificus during cold storage

Different cultural techniques and molecular methods for the detection of Vibrio vulnificus during cold storage in a model broth system were compared. Two strains of V. vulnificus were grown to stationary phase and inoculated (10(6) CFU/mL) into tryptic soy broth with 2% sodium chloride (TSBN2) or artificial seawater (ASW), both pre-chilled to 5 degrees C. These were stored for 10 days, with sub-sampling conducted at time 0 and every 2 days thereafter. Each subsample was plated, by both pour and spread plate techniques, onto tryptic soy agar 2% sodium chloride (TSAN2) with or without catalase (400 or 600 U) or sodium pyruvate (80 or 160 mg) supplementation. Nucleic acids were extracted from subsamples and subjected to PCR and RT-PCR with hemolysin as the target. Higher recoveries of V. vulnificus were obtained with spread plating compared to pour plating (P<0.05). The addition of sodium pyruvate (80 mg) or catalase (400 U) significantly increased cell recovery (P<0.05). PCR amplification signals were stronger than RT-PCR signals at each timepoint, and results were generally consistent between TSAN2 and ASW for each strain. These results will aid in the design of optimum methods to recover and/or detect V. vulnificus cells subjected to sublethal stress that might be encountered in food processing and storage.     

Use of mutant strain for evaluating processing strategies to inactivate Vibrio vulnificus in oysters.

Vibrio vulnificus is a ubiquitous marine bacterium frequently isolated from shellfish and associated with severe and often fatal disease in humans. Various control strategies to reduce the disease risk associated with V. vulnificus contamination in shellfish have been proposed. However, evaluating the efficacy of these control strategies is complicated because of the difficulty in distinguishing V. vulnificus from the high levels of background environmental Vibrio spp. The purpose of this research was to develop a model indicator V. vulnificus strain that could be readily differentiated from background microflora and used to facilitate the evaluation of processing efficacy. A spontaneous nalidixic acid-resistant strain of V. vulnificus (Vv-NA) was prepared from a wild-type parent (Vv-WT) using selective plating techniques. Vv-NA was very similar to Vv-WT with respect to biochemical characteristics, appearance on selective plating media, detection limits using most probable number and polymerase chain reaction, and growth rate. In comparative freeze inactivation studies on pure cultures, Vv-WT and Vv-NA had similar freeze inactivation profiles at -20 degrees C (conventional freezing), at -85 degrees C (cold blast freezing), and in liquid nitrogen (cryogenic freezing). In oyster homogenates artificially inoculated with Vv-NA, the organism was inactivated 95 to 99% after freezing, irrespective of freezing temperature. Thermal inactivation comparisons of pure cultures of Vv-WT and Vv-NA using the capillary tube method revealed statistically significant differences in D values at 47 degrees C (2.2 versus 3.0 min, respectively) and 50 degrees C (0.83 versus 0.56 min, respectively), but nearly identical values at 52 degrees C (0.21 versus 0.22 min, respectively). However, these D values were notably higher than those reported by other investigators and hence provided a conservative means by which to evaluate thermal inactivation. In oyster homogenates seeded with Vv-NA, D values of 1.3+/-0.09 min and 0.41+/-0.01 min were obtained at 46 degrees C and 48 degrees C, respectively. This study demonstrated that Vv-NA is readily enumerated and could be used as a surrogate for evaluating the degree of V. vulnificus inactivation provided by freezing and thermal treatments of oyster homogenates.     

Effect of pH, NaCl content, and temperature on growth and survival of Arcobacter spp

Growth and survival of six human isolates of the pathogenic Arcobacter spp. in the presence of selected environmental factors were studied. Four strains of Arcobacter butzleri and two strains of Arcobacter cryaerophilus were exposed to pH levels of 3.5 to 8.0. Most strains grew between pH 5.5 and 8.0, with optimal growth of most A. butzleri and A. cryaerophilus strains at pH 6.0 to 7.0 and 7.0 to 7.5, respectively. The 24-h optimal growth range in the presence of NaCl was 0.5 to 1.0% for A. cryaerophilus. However, after 96 h, the optimum was between 0.5 and 2.0% NaCl. The optimum range for growth of A. butzleri strains was 0.09 to 0.5% NaCl after 96 h. The upper growth limits were 3.5 and 3.0% NaCl for A. butzleri and A. cryaerophilus, respectively. Survival at 25 degrees C in up to 5% NaCl was noted for A. butzleri 3556 and 3539 and A. cryaerophilus 3256. Decimal reduction times (D-values) at pH 7.3 in phosphate-buffered saline for three A. butzleri strains were 0.07 to 0.12 min at 60 degrees C, 0.38 to 0.76 min at 55 degrees C, and 5.12 to 5.81 min at 50 degrees C. At pH 5.5, decreased thermotolerance was observed, with D-values of 0.03 to 0.11 min at 60 degrees C, 0.30 to 0.42 min at 55 degrees C, and 1.97 to 4.42 min at 50 degrees C. Calculated z-values ranged from 5.20 to 6.28 degrees C. D-values of a three-strain mixture of A. butzleri in raw ground pork were 18.51 min at 50 degrees C and 2.18 min at 55 degrees C. Mild heat (50 degress C) followed by cold shock (4 or 8 degrees C exposure) had a synergistic lethal effect, reducing more cells than with an individual 50 degrees C treatment or with cold shock temperatures of 12 or 16 degrees C.     

Virulence and stress susceptibility of clinical and environmental strains of Vibrio vulnificus isolated from samples from Taiwan and the United States

Vibrio vulnificus is an estuarine bacterium that causes severe wound infection and septicemia with high mortality. It also can be transmitted through the consumption of raw contaminated seafood and is an important foodborne pathogen. A total of 40 environmental and clinical V. vulnificus strains isolated from the United States and Taiwan were analyzed for virulence in animals, the presence of virulence-associated factors, and susceptibility to environmental stresses. Virulence in mice was exhibited by 85% of the environmental strains and 95% of the clinical strains. Strains from environmental or clinical sources were similar in virulence-associated phenotypes (protease activity, utilization of transferrin-bound iron, hemolysis, and inactivation in serum) and susceptibility to various stresses (4 and 52 degrees C, 0.1 and 10% NaCl, and pH 3.2), except freeze-thaw treatment. The clinical strains killed experimental animals after a shorter incubation time than did the environmental strains. Most of the 15 virulence-associated genes examined were present in most of the strains, regardless of their sources or virulence, with the exception of vvh, flgF, and purH. vvh was significantly more common in clinical strains than in environmental strains, and vvh, flgF, and purH were more common in virulent strains than in nonvirulent strains. These data may be helpful in devising strategies to manage or reduce the presence of V. vulnificus in foods.     

广州地区海产品中创伤弧菌病原学特征分型

目的研究广州地区海产品中分离的创伤弧菌的毒力相关基因与脉冲场凝胶电泳(PFGE)分子分型状况,初步建立区域PFGE溯源数据库,为该菌的分子流行病学研究和食源性疾病溯源提供支持。方法对68株创伤弧菌进行毒力相关基因鉴定和PFGE分型,采用Bio Numerics软件分析聚类关系。结果海产品中分离的创伤弧菌均为溶血素A基因(vvh A)核酸阳性,vcg C/E和16S rRNA A/B分型结果显示,共有5种基因型别,分别有CB型、EA型、CAB型、CA型和EB型。共检测出42株为BT1型。PFGE分型结果显示68株创伤弧菌有52种PFGE型别。结论广州地区海产品中分离的创伤弧菌PFGE型别多,且相互之间关系分散;毒力相关基因型以CB型为主。     

Characterization of the low-salinity stress in Vibrio vulnificus

Vibrio vulnificus is a marine pathogenic bacterium commonly found in seawater or seafood. This organism encounters low-salinity stress in its natural environment and during food processing. This study was designed to investigate the response of V. vulnificus YJ03 to lethal low salinity (0.04% NaCl) and its adaptation to sublethal salinity (0.12% NaCl with 20 amino acids added). A short period in the nonculturable state was induced by lethal low-salinity stress followed by cell death after 30 min of stress. Addition of 1 mM glycine betaine or 0.5 mM sucrose reduced the damage. Low-salinity adaptation was achieved in the exponential-phase cells but not in the stationary-phase cells. Significant protection against lethal low-salinity stress was attained when the cells were adapted for as little as 1.5 min. The adapted cells were significantly protected against lethal low salinity and 2.4% sodium sorbate but sensitized to the challenge of heat (52 degrees C) and acid (pH 3.2). Nonlethal low-salinity treatment of seafood should be avoided to prevent stress adaptation of V. vulnificus.     

Response of Vibrio parahaemolyticus 03:K6 to a hot water/cold shock pasteurization process.

Vibrio vulnificus and V. parahaemolyticus are natural inhabitants of estuarine environments world wide. Pathogenic strains of these bacteria are often transmitted to humans through consumption of raw oysters, which flourish in the same estuaries. Previous studies reported the effective use of hot water pasteurization followed by cold shock to eliminate from raw oysters naturally and artificially incurred environmental strains of V. vulnificus and V. parahaemolyticus common to the Gulf of Mexico. The present study focused on the use of the same pasteurization method to reduce a highly process resistant Vibrio strain, V. parahaemolyticus O3:K6 to non-detectable levels. Oysters were artificially contaminated with 10(4) and 10(6) V. parahaemolyticus 03:K6 cfu g(-1) oyster meat. Contaminated oysters were pasteurized between 50 and 52 degrees C for up to 22 min. Samples of processed oysters were enumerated for V. parahaemolyticus O3:K6 at 2-min intervals beginning after the 'come-up time' to achieve an oyster internal temperature of at least 50 degrees C. The D value (D(52)deg C) was 1.3-1.6 min. V. parahaemolyticus O3:K6 proved more process resistant than non-pathogenic environmental strains found in Gulf of Mexico waters. A total processing time of at least 22 min at 52 degrees C was recommended to reduce this bacterium to non-detectable levels (< 3 g(-1) oyster meat).     

Heat resistance of juice spoilage microorganisms

The heat resistance of various yeasts (Saccharomyces cerevisiae, Rhodotorula mucilaginosa, Torulaspora delbrueckii, and Zygosaccharomyces rouxii), molds (Penicillium citrinum, Penicillium roquefortii, and Aspergillus niger), and lactic acid bacteria (Lactobacillus fermentum and Lactobacillus plantarum) obtained from spoiled acid or acidified food products was determined in 0.1 M citrate buffer at pH values of 3.0, 3.5, and 4.0. S. cerevisiae was the most heat resistant of the microorganisms in citrate buffer, and its heat resistance was further evaluated in apple, grapefruit, calcium-fortified apple, and tomato juices as well as in a juice base with high fructose corn syrup. Decimal reduction times (D-values) and changes in temperature required to change the D-value (z-values) for S. cerevisiae were higher in the juices than in citrate buffer at all pH values tested. The D57 degrees C(135 degrees F)-values varied from 9.4 min in the juice product with pH 2.8 to 32 min in a calcium-added apple juice with pH 3.9. The S. cerevisiae strain used in this study can be used in thermal-death-time experiments in acidic products to calculate process conditions and in challenge tests to validate the calculated temperatures and hold times during processing.     

Susceptibility of the heat-, acid-, and bile-adapted Vibrio vulnificus to lethal low-salinity stress

As a marine pathogenic bacterium that inhabits seawater or seafood, Vibrio vulnificus encounters low salinity and other stresses in the natural environment and during food processing. This investigation explores the cross-protective response of sublethal heat-, acid-, or bile-adapted V. vulnificus YJ03 against lethal low-salinity stress. Experimental results reveal that the acid (pH 4.4)- and heat (41 degrees C)-adapted V. vulnificus were not cross-protected against the lethal low-salinity challenge (0.04% NaCl). The bile (0.05%)-adapted exponential- and stationary-phase cells were cross-protected against low salinity, whereas low-salinity (0.12% NaCl)-adapted stationary cells were sensitized against 12% bile stress. Results of this study provide further insight into the interaction between low salinity and other common stresses in V. vulnificus.     

Relationship between the effects of stress induced by human bile juice and acid treatment in Vibrio cholerae

The effects of low pH and human bile juice on Vibrio cholerae were investigated. A mild stress condition (exposure to acid shock at pH 5.5 or exposure to 3 mg of bile per ml for 20 min) slightly decreased (by < or = 1 log unit) V. cholerae cell viability. However, these treatments induced tolerance to subsequent exposures to more severe stress. In the O1 strain, four proteins were induced in response to acid shock (ca. 101, 94, 90, and 75 kDa), whereas only one protein (ca. 101 kDa) was induced in response to acid shock in the O139 strain. Eleven proteins were induced in response to bile shock in the O1 strain (ca. 106, 103, 101, 96, 88, 86, 84, 80, 66, 56, and 46 kDa), whereas only one protein was induced in response to bile shock in the O139 strain (ca. 88 kDa). V. cholerae O1 and O139 cells that had been preexposed to mild acid shock were twofold more resistant to pH 4.5 (with times required to inactivate 90% of the cell population [D-values] of 59 to 73 min) than were control cells (with D-values of 24 to 27 min). Likewise, cells that were preexposed to mild bile shock (3 mg/ml) were almost twofold more tolerant of severe bile shock (30 mg/ml; D-values, 68 to 87 min) than were control cells (with D-values of 37 to 43 min). These protective effects persisted for at least 1 h after the initial shock but were abolished when chloramphenicol was added to the culture during the shock. Cells preexposed to acid shock exhibited cross-protection against subsequent bile shock. However, cells preexposed to bile shock exhibited no changes in acid tolerance. Bile shock induced a modest reduction (0 to 20%) in enterotoxin production in V. cholerae, whereas acid shock had no effect on enterotoxin levels. Adaptation to acid and bile juice and protection against bile shock in response to preexposure to acid shock would be predicted to enhance the survival of V. cholerae in hosts and in foods. Thus, these adaptations may play an important role in the development of cholera disease.     

PROTEIN PROFILE CHANGES IN ACID ADAPTED LISTERIA MONOCYTOGENES EXHIBITING CROSS-PROTECTION AGAINST AN ACTIVATED LACTOPEROXIDASE SYSTEM IN TRYPTIC SOY BROTH

Foodborne pathogens often tolerate and survive environmental stress conditions including extreme acidity to varying degrees. One possible reason for this survival may be the production of protective stress proteins during acid shock (ASR) and/or tolerance (ATR) responses. The ASR and ATR of Listeria monocytogenes strains V7, V37 and CA in tryptic soy broth without dextrose acidified with lactic acid were studied. Possible cross-protection of acid adapted cells against an activated lactoperoxidase system was also determined. The strains were either directly challenged at pH 4.0 and 3.5 to study their ASR or initially adapted at pH 5.5 for the equivalent of 1 generation before challenging at pH 4.0 and 3.5 to study their ATR. Adapted and nonadapted cells were challenged at pH 4.5 with or without an activated lactoperoxidase system. In all cases viability was determined by enumeration over a period of 24 or 48 h after challenge and the production of stress proteins analyzed by 2-dimensional gel electrophoresis. While there were some differences in the survival responses for each strain, the acid adapted cells of each strain survived to a greater degree than nonadapted cells at both pH 4.0 (at least 10 fold at 24 h) and pH 3.5 (at least 1000 fold at 6 h) but not at pH 4.5. The acid adapted cells exposed to the lactoperoxidase system survived better (at least 5-fold) than their nonadapted counterparts for all 3 strains at 24 and 48 h. The 2-dimensional gel analysis for all 3 strains showed that the adapted and nonadapted cells underwent a change in their physiology, (at pH 4.0 compared to the control at pH 7.0; at pH 4.5 with the addition of lactoperoxidase system components) in that there was induction as well as repression of several proteins.     

Detection and Enumeration of Vibrio vulnificus by Direct Colony Immunoblot

ABSTRACT:  A direct colony immunoblot method (DCI) for the enumeration of Vibrio vulnificus was developed. Bacterial colonies were transferred from agar plates to membranes, which were then dried and blocked with bovine serum albumin. Subsequently, the membranes were treated with anti- V. vulnificus H antibodies, washed and incubated with peroxidase-conjugated goat anti-rabbit IgG. After a final wash, the membranes were exposed to a substrate mixture containing H₂O₂ which resulted in the development of a purple color by V. vulnificus colonies. The DCI detected all clinical and environmental V. vulnificus strains tested and did not cross-react with other Vibrio species including V. cholerae , V. parahaemolyticus , or V. fluvialis . The DCI was then compared to the DNA hybridization procedure (DNAH) using V. vulnificus agar plates inoculated with mixed cultures of V. vulnificus and V. parahaemolyticus and V. vulnificus -seeded oyster homogenates. Both DCI and DNAH detected 1 to 2 log colony forming units (CFU)/mL V. vulnificus mixed with 4 log CFU/mL V. parahaemolyticus . Both methods were comparable and demonstrated no significant statistical differences when enumerating V. vulnificus in mixed cultures or in oyster homogenates seeded with levels of V. vulnificus from 2 to 6 log CFU/mL. The DCI demonstrated clearer color development and was less time consuming than the DNAH.     

Temperature Effects on the Depuration of Vibrio parahaemolyticus and Vibrio vulnificus from the American Oyster (Crassostrea virginica)

ABSTRACT:  This study investigated temperature effects on depuration for reducing Vibrio parahaemolyticus and Vibrio vulnificus in American oyster ( Crassostrea virginica ). Raw oysters were inoculated with 5-strain cocktail of V. parahaemolyticus or V. vulnificus to levels of 10⁴ to 10⁵ MPN (most probable number)/g and depurated in artificial seawater (ASW) at 22, 15, 10, and 5 °C. Depuration of oysters at 22 °C had limited effects on reducing V. parahaemolyticus or V. vulnificus in the oysters. Populations of V. parahaemolyticus and V. vulnificus were reduced by 1.2 and 2.0 log MPN/g, respectively, after 48 h of depuration at 22 °C. Decreasing water temperature to 15 °C increased the efficacy of depuration in reducing V. parahaemolyticus and V. vulnificus in oysters. Reductions of V. parahaemolyticus and V. vulnificus in oysters increased to 2.1 and 2.9 log MPN/g, respectively, after 48 h of depuration at 15 °C. However, depurations at 10 and 5 °C were less effective than at 15 °C in reducing the Vibrio spp. in oysters. Extended depuration at 15 °C for 96 h increased reductions of V. parahaemolyticus and V. vulnificus in oysters to 2.6 and 3.3 log MPN/g, respectively.     

Thermal resistance of spores from virulent strains of Bacillus anthracis and potential surrogates

The objective of this study was to determine the thermal resistance of spores of Bacillus anthracis and potential surrogates. The heat resistance of spores suspended in buffer (pH 7.0 or 4.5), milk, or orange juice was determined at 70, 80, and 90 degrees C. D-values for B. anthracis strains Sterne, Vollum, and Pasteur ranged from < 1 min at 90 degrees C to approximately 200 min at 70 degrees C and were lower under acidic than under neutral conditions. The D-values for B. anthracis spores fell within the range obtained for spores from eight strains of Bacillus cereus, Bacillus thuringiensis, Bacillus mycoides, and Bacillus subtilis. However, there were significant differences (P < 0.001) among the D-values of the strains. The z-values in pH 7.0 buffer and milk averaged approximately 10.5 degrees C and were not significantly different among strains (P < 0.05). The z-values in pH 4.5 buffer and orange juice averaged 12.9 and 13.9 degrees C, respectively, significantly (P < 0.05) higher than those obtained in milk or in pH 7.0 buffer. The significance of this difference was driven by large differences among a few strains. The z-values for B. anthracis strain Pasteur were twice as high in the acid media than in the neutral media. This study confirms that B. anthracis spores are not unusually heat resistant and that spores from validated Bacillus species are appropriate surrogates for thermal resistance studies.     

酸胁迫对鼠伤寒沙门氏菌抗酸性、细胞膜及膜蛋白的影响

目的：以鼠伤寒沙门氏菌CGMCC 1.1190为对象，研究柠檬酸反复胁迫处理对其抗酸性、细胞膜及其膜蛋白的影响。方法：将CGMCC 1.1190分别在经柠檬酸调节到pH值为3.0、2.7、2.5的胰蛋白胨大豆肉汤培养基中胁迫处理并转接12 次培养后，获得3 株CGMCC 1.1190的抗酸性菌株，测定了这3 株抗酸性菌株的D值、菌落形态、个体形态、膜通透性、膜流动性和膜蛋白的变化。结果：这3 株抗酸性菌株的D值随着酸处理次数的增加不断增大，酸胁迫的pH值越低，D值越大，菌落形态与个体形态变化越明显；与原始对照菌株相比，当这3 株抗酸性菌株置于pH 2.0的强酸环境下时，其碘化丙啶染色区域的死菌比例显著降低（P＜0.05），表明酸胁迫pH值越低，这3 株抗酸性菌株的细胞膜对H＋通透性越低；随着酸胁迫pH值的降低，CGMCC 1.1190抗酸性菌株细胞膜磷脂的熔点升高，表明细胞膜磷脂的相变温度升高，细胞膜流动性降低；十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结果表明，这3 株抗酸性菌株分子质量35～180 kDa范围内的细胞膜蛋白表达量增加，在135 kDa和180 kDa处均增加了特异条带。结论：随着酸胁迫pH值的降低，CGMCC 1.1190抗酸性增加，菌落变小，个体形态变长，细胞膜通透性和流动性均降低，部分膜蛋白表达量和表达种类增加，这些变化有利于其适应在酸胁迫环境下生长，提高生存能力。