Recovery of Escherichia coli ATCC 25922 in phosphate buffered saline after treatment with high hydrostatic pressure

Escherichia coli ATCC 25922 in phosphate buffered saline solution (PBS, pH 7.1, 10(8) CFU/ml) was inactivated by high hydrostatic pressure (HHP, 400 to 600 MPa) treatment at 25 degrees C for 10 min. Colonies of E. coli were not detected on non-selective plate count agar immediately after a HHP-treatment of at least 550 MPa. E. coli subjected to at least 500 MPa in PBS were incubated at 4, 25, and 37 degrees C for 120 h. No colonies were detected on plate count agar throughout the 120-h incubation period at 4 or 37 degrees C. In contrast, the number of E. coli during storage at 25 degrees C increased from an undetectable level (< 1 CFU/ml) to the level of initial cell counts regardless of the treatment pressure level. The recovery in PBS required a maximum time of 48 h, while the period during which cell numbers remained at an undetectable level increased from 24 to 72 h as the treatment pressure increased. E. coli treated at 550 and 600 MPa in PBS were inoculated into trypticase soy broth (TSB) and stored at 4, 25, and 37 degrees C for 120 h. No recovery was recorded in TSB during the 120-h storage at 37 degrees C. In contrast, the number of E.coli during storage at 25 degrees C in TSB increased beyond the level of initial cell counts regardless of the treatment pressure level. The recovery of cell numbers observed in TSB was faster than that in PBS samples, as bacterial growth in TSB assisted faster recovery. When the incubation temperature in PBS was shifted to 25 degrees C after 120-h at 4 or 37 degrees C, recovery of E. coli was observed in samples shifted from 4 to 25 degrees C regardless of the treatment pressure. However, the time during which cell numbers remained at an undetectable level was extended by increasing the level of treatment pressure, and recovery required a maximum time of 48 h. On the other hand, no recovery was observed with HHP-treated E. coli subjected to an incubation temperature shift from 37 to 25 degrees C. This study indicates that an appropriate incubation temperature after HHP-treatment is needed to optimize the recovery of HHP-injured bacteria and thus prevent overestimation of the lethal effect of HHP-treatment.     

The enumeration of coliforms and E. Coli on naturally contaminated beef: A comparison of the petrifilm(™) method with the Australian Standard

Petrifilm(?) PEC was compared to Australian Standard (AS) methods for the enumeration of coliforms and Escherichia coli on 50 naturally contaminated beef samples from three meat works. The standard methods consist of a 3-tube most probable number test or a direct plate count on tryptone bile agar for E. coli, and violet red bile agar and the most probable number test for conliorms (AS 1766.2.12.1984 and AS 1766.2.3.1992). No significant difference was found between the methods, except that the count of E. coli on the direct plate method with a resuscitation step was significantly higher than the count on Petrifilm(?), most probable number or direct plate count without resuscitation.     

High-performance hybrid composites made of recycled Nomex,Kevlar, and polyester selvages: mechanical property evaluations

In modern age, prosperous business, industrial, social, and leisure activities improve and ensure the quality of lives considerably, but are inevitably accompanied with insecurity and dangers that lurk anywhere anytime. People are concerned about safety and have increasing demands on self-protection in terms of working/living environment, accidents, and raising crime rates. The increasing demands of protective textiles cause the mass production of high performance fabrics, leaving considerable amount of high performance selvages. In this study, three kinds of high performance recycled selvages, including Nomex, Kevlar, and PET selvages, are used to make high-performance hybrid composites. The recycled selvages are smashed into staple fibers using a nonwoven manufacturing process. Each recycled staple fibers are individually combined with low-melting-point polyester (LPET) fibers at ratios of 9:1, 7:3, and 5:5, and processed with thermal compression. The LPET fibers contribute to the hybrid composites with thermal bonding points, which decrease the voids between fibers, limit the fiber slide, and enhance the friction force between fibers. The shearing, expelling, and friction functionalities of LPET fibers are expected have a positive influence on the mechanical properties of the high-performance hybrid composites. The morphology of hybrid composites is observed using a scanning electron microscope, and the air permeability, tensile strength, tearing strength, and bursting strength of hybrid composites are evaluated, examining the optimal parameters.     

利用EHEC琼脂平板从食物中分离检测产志贺毒素大肠杆菌

为利用肠出血性大肠杆菌(EnterohaemorrhagicEscherichiacoli,EHEC)琼脂平板,从食物中分离、检测产志贺毒素大肠杆菌(shigatoxin-producingEscherichiacoli,STEC),采用STEC毒力基因(hlyA基因)检测溶血素的产生,建立了一种利用EHEC琼脂平板快速、准确地从食物中分离、检测STEC的方法。该方法可检测STEC不同血清O157∶H7、O26、O111。     

Penetration of Escherichia coli O157:H7 into lettuce tissues as affected by inoculum size and temperature and the effect of chlorine treatment on cell viability

Penetration of Escherichia coli O157:H7 into iceberg lettuce tissues and the effect of chlorine treatment on cell viability were evaluated. Attachment of different inoculum levels (10(9), 10(8), and 10(7) CFU/ml) was examined by determining the number of cells at the surface and the cut edge of lettuce leaves (2 by 2 cm). E. coli O157:H7 attached preferentially to cut edges at all inoculum levels, with greater attachment per cm2 of lettuce at higher inoculum levels. A longer attachment time allowed more cells to attach at both sites. Immunostaining with a fluorescein isothiocyanate-labeled antibody revealed that cells penetrated into lettuce leaves from cut edges. Cells showed greater penetration when lettuce was held at 4 degrees C compared with 7, 25, or 37 degrees C and were detected at an average of 73.5 +/- 16.0 microm below the surfaces of cut tissues. Penetrating cells were mostly found at the junction of lettuce cells. The viability of attached cells after treatment with 200 mg/liter (200 ppm) of free chlorine for 5 min was examined by plating on tryptic soy agar and by a nalidixic acid elongation method. Although chlorine treatment caused significant reduction in attachment (0.7- and 1.0-log reduction at surfaces and cut edges, respectively), cells remained attached at high numbers (7.9 and 8.1 log CFU/cm2 at surfaces and cut edges, respectively). Elongated cells were observed in stomata and within the tissues of the lettuce, indicating they were protected from contact with chlorine.     

Detection of Enterotoxigenic Escherichia coil in Foods by DNA Colony Hybridization

A DNA colony hybridization procedure was used to identify and enumerate a heat-labile toxin-producing strain of Escherichia coli (H10407) in various types of foods. Foods were seeded with H10407 cells and examined by DNA hybridization on nitrocellulose filters with ³²P- labeled heat-labile toxin gene fragments. The number of cells recovered on plate count agar and eosine methylene blue agar was compared. With nitrocellulose filters, recoveries were about 81% on plate count agar and about 76% on eosine methylene blue agar. No significant differences in recovery were observed with ten different food types.     

天津主要河流中耐高温蛭弧菌的分离及生物学特性

从天津4条主要河流(独流减河,南运河,,北运河,子牙河)中分离出了耐高温的蛭弧菌并进行了纯化.通过采用宿主双层琼脂平板法对从河水中筛选出耐高温菌株并对其进行宿主范围、pH、温度等生物学特性的测定,结果表明:耐高温蛭弧菌在pH 7.0～7.5生长最好;对热致死的大肠杆菌,嗜水气单胞菌,荧光假单胞菌等3种细菌仍有裂解作用且...     

EVALUATION OF A RIBOFLAVIN-METHIONINE MIXTURE TO IMPROVE THE QUALITY OF AQUACULTURED CATFISH FILETS DURING THE CHILLING PROCESS

An optimized riboflavin-methionine (RM) mixture with Escherichia coli (target organism) was evaluated for improving the quality of catfish filets during the chilling process. The RM mixture (pH 7.2) containing riboflavin (10μM), methionine (20 mM) and EDTA (1 mM) was photoactivated. E. coli cells grew in a medium containing l-methionine when exposed to fluorescent light and grew in a RM mixture in the dark. However, the activated RM mixture injured and killed E. coli cells. As the concentration of riboflavin increased, the number of survivors decreased. The concentration of methionine did not significantly decrease the survival rate of E. coli. As pH increased, the number of killed and injured E. coli cells increased. The RM mixture injured and/or killed a low load of E. coli cells faster than a high load. Methionine solution or RM mixture added to the suspension in phosphate buffer (15 mM, pH 7.2) and photoactivated at 0 and 24C for 8h to simulate the chilling water significantly reduced the total coliform counts but not the standard plate counts. However, when the catfish filets were suspended in a RM mixture at 0 and 24C for 2h to simulate the chilling process, there was no reduction in standard plate or total coliform counts.     

聚乙烯/聚酯纤维卷曲对热风非织造材料性能的影响

为获得蓬松、棉柔兼具丝柔爽滑特点的卫生用热风非织造材料,选取皮芯结构的聚乙烯/聚酯纤维为原料,通过控制生产工艺改变纤维的卷曲度,制备20 g/m^2的不同结构的热风非织造材料。通过对热风非织造材料性能的测试分析,研究聚乙烯/聚酯纤维的卷曲度对热风非织造材料的机械、透气、透湿、渗透、回渗、摩擦性能的影响。结果表明:聚乙烯/聚酯纤维的卷曲度降低,制得的热风非织造材料的液体渗透速率、摩擦性能均有所提高,机械、透气、透湿性能基本不变。当聚乙烯/聚酯纤维的卷曲度为12.08%时,制得的热风非织造材料的渗透时间为0.61 s,回渗量为0.04 g,试样横、纵向摩擦因数分别为0.217、0.190,其透湿、渗透、摩擦性能均优于同面密度下的纺粘非织造材料,可应用于一次性卫生用品面层材料。     

海藻酸铜纤维的抗菌及抑菌性能

为研究海藻酸铜纤维的抗菌及抑菌性能,以大肠埃希氏杆菌、铜绿假单胞菌、金黄色葡萄球菌、枯草芽孢杆菌为实验菌株,采用琼脂平皿扩散法和烧瓶振荡法对海藻酸铜纤维的抗菌性能进行研究和评价,并借助扫描电子显微镜技术对其抗菌机制进行分析.结果表明:海藻酸铜纤维对革兰氏阴性菌和革兰氏阳性菌均具有良好的抗菌性;细菌与纤维接触后由于铜离子...     

Survival of Escherichia coli O157:H7 in frozen foods: impact of the cold shock response

The survival of Escherichia coli O157:H7 strains in both frozen foods and trypticase soy broth (TSB) was investigated following cold shocking at 10 degrees C for 1.5 h. Using both trypticase soy agar (TSA) and violet red bile agar (VRBA) as recovery media, it was demonstrated that survival levels between cold shocked (CS) and non-cold shocked (NS) E. coli in ground beef or pork were not significantly different (P < or = 0.05). In contrast, cold shocking E. coli in either milk, whole egg or sausage resulted in a significant(P < or = 0.05) enhancement in survival. For milk, survival levels of CS E. coli, by 28 days of frozen storage, were 1.89 and 1.66 log10 cfu/ml higher on TSA and VRBA, respectively, when compared to NS cells. In egg these values were 0.64 and 1.31, while in sausage, values of 0.74 and 1.19 were obtained. In TSB (pH 7) survival of CS E. coli for some strains was about 3 log10 cfu/ml higher when compared to NS cells. Acidification of TSB (pH 5), however, appeared to negate the protective effects of the cold shock treatment. In milk, increasing the differential between the growth and cold shock temperatures resulted in higher numbers of survivors. In this regard the growth-cold shock temperature protocol giving optimum protection was 37-10 degrees C. In contrast, growth of E. coli at 20 degrees C followed by cold shocking at 10 degrees C did not result in any significant freeze protection. In addition, increased protection due to cold shocking was correlated with the appearance of a novel protein appearing at pI 4.8 following isoelectric focusing analysis, thus demonstrating an alteration of protein synthesis.     

Penetration of Escherichia coli O157:H7 into lettuce as influenced by modified atmosphere and temperature.

The effects of temperature and atmospheric oxygen concentration on the respiration rate of iceberg lettuce and Escherichia coli O157:H7 cells attachment to and penetration into damaged lettuce tissues were evaluated. Respiration rate of lettuce decreased as the temperature was reduced from 37 to 10 degrees C. Reducing the temperature further to 4 degrees C did not affect the respiration rate of lettuce. Respiration rate was also reduced by lowering the atmospheric oxygen concentration. Lettuce was submerged in E. coli O157:H7 inoculum at 4, 10, 22, or 37 degrees C under 21 or 2.7% oxygen. Attachment and penetration of E. coli O157:H7 were not related to the respiration rate. The greatest numbers of E. coli O157:H7 cells attached to damaged lettuce tissues at 22 degrees C at both oxygen concentrations. More cells were attached under 21% oxygen than under 2.7% oxygen at each temperature, but this difference was small. Penetration of E. coli O157:H7 into lettuce tissue was determined by immunostaining with a fluorescein isothiocyanate-labeled antibody. Under 21% oxygen, E. coli O157:H7 cells showed greatest penetration when lettuce was held at 4 degrees C, compared to 10, 22. or 37 degrees C, and were detected at an average of 101 microm below the surfaces of cut tissues. However, under 2.7% oxygen, there were no differences in degree of penetration among four incubation temperatures. The degree of E. coli O157:H7 penetration into lettuce tissue at 4 or 22 degrees C was greater under 21% oxygen than under 2.7% oxygen; however, no difference was observed at 37 degrees C. Conditions that promote pathogen penetration into tissue could decrease the effectiveness of decontamination treatments.     

Study of stiffness and abrasion resistance of needle‐punched nonwoven blankets

The properties of needled fabrics depend on the nature of component fibers and the manner in which fibers are arranged in the structure. Fiber properties along with the various machine and web parameters contribute to the structure that emerges from the needling operation. In this paper, the effect of machine parameters such as depth of needle penetration and punch density on fabric stiffness and abrasion resistance of needled blankets has been studied. The effect of calendering and sandwiching hollow polyester fibers between two layers of fine polyester fibers on the abrasion resistance and fabric stiffness has also been studied. It was observed that fabric stiffness first increases and then decreases as the depth of needle penetration increases. Increase in punch density leads to a decrease in fabric stiffness only at higher levels of depth of penetration due to fiber rupture. Calendering improves the fabric abrasion resistance properties but fabric stiffness also increases. Sandwiching of hollow polyester fibers between the two layers of fine denier polyester fibers improves the abrasion resistance without increasing the fabric stiffness.     

碳化硅纤维预制体编织损伤特性研究

针对编织过程中碳化硅(SiC)纤维束的摩擦磨损会导致其可编织性下降等问题,在传统抱合力机基础上通过自制载荷可控的抱合夹具,模拟SiC纤维束在编织过程中与机械部件之间的摩擦行为,研究了法向载荷、摩擦速度和摩擦次数对SiC纤维束磨损行为的影响。研究表明:SiC纤维拉伸断裂表现出明显的脆性断裂行为,SiC纤维束的摩擦断裂循环次数随着法向载荷和摩擦速度的增加而显著减少,拉伸强度和断裂伸长率随着摩擦次数的增加而明显降低;当摩擦次数增加到100次时,SiC纤维束的拉伸断裂强力和断裂伸长率较原样分别减小了73%和53%;随着摩擦次数的增加,SiC纤维束破坏主要经历纤维分散、纤维起毛和纤维断裂等损伤过程。     

Evaluation of a rapid fluorogenic method for the detection of Escherichia coli in dairy products.

A rapid fluorogenic medium was evaluated for the detection of Escherichia coli in dairy products. The medium was capable of detecting Esch. coli after 7.5 h incubation at 41.5 degrees C. Samples of pasteurized milk (136), raw milk (63), soft cheese (60) and pasteurized cream (39) were examined with media based on 4-methylumbelliferyl-beta-D-glucuronide (MUG-7) and Violet red bile agar and there were no significant differences between the numbers of Esch. coli detected on the two media. MUG-7 medium had a specificity of 98.6% and the small number of organisms giving a false positive reaction were identified as Klebsiella pneumoniae. The incidence of false negative results was approximately 2%. MUG-7 medium was suitable for pour plate, spread plate and membrane filtration methods. Possible applications of the method are discussed.     

Microbial evaluation of selected fresh produce obtained at retail markets

The microbial quality of five types of fresh produce obtained at the retail level was determined by standard quantitative techniques. These techniques included aerobic plate count (APC), total coliform counts, Escherichia coli counts, and yeast and mold counts. Three different methods were used to determine total coliform counts, which consisted of MacConkey agar plate counts, Colicomplete most probable number counts, and Petrifilm E. coli (EC) plate counts. The mean APCs for sprouts, lettuce, celery, cauliflower, and broccoli were 8.7, 8.6, 7.5, 7.4. and 6.3 log10 CFU/g, respectively. MacConkey agar counts indicated that 89 to 96% of the APCs consisted of gram-negative bacteria. Yeast and mold counts were in a range expected of fresh produce. Fresh produce was also analyzed for human pathogens. Samples were analyzed for Staphylococcus spp., Bacillus spp., Salmonella spp., Listeria spp., and Campylobacter spp. One isolate of Staphylococcus was found to be enterotoxigenic, and one species of Bacillus was also toxigenic. Neither Salmonella spp. nor Campylobacter spp. were detected in any of the produce samples. A variety of Listeria spp., including Listeria monocytogenes, were found in fresh produce.     

南五味子提取物对食品常见致病菌抑制作用的研究

探讨南五味子提取物对食品常见致病菌的抑制作用。采用乙醚、70%乙醇和水3种提取剂对南五味子进行提取,以大肠杆菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)和乙型副伤寒沙门氏菌(Salmonella paratyphi B)为供试菌种,通过琼脂平板扩散法测定抑菌圈;琼脂平板二倍稀释法测定最低抑菌浓度(MIC,g/mL)。3种提取物的抑菌效果大小为:70%乙醇提取物>水提取物>乙醚提取物。70%乙醇提取物抑菌效果最好,对大肠杆菌、金黄色葡萄球菌和沙门氏菌的抑菌圈分别达到26.4,25.2,23.3 mm,MIC分别为0.0125,0.0125,0.025 g/mL。南五味子提取物对食品常见致病菌的抑制作用显著,可开发成安全有效的天然食品防腐剂。     

Influence of punctures, cuts, and surface morphologies of golden delicious apples on penetration and growth of Escherichia coli O157:H7

The ability of Escherichia coli O157:H7 to penetrate and grow within punctures, fresh-cut surfaces, and calyces of Golden Delicious apples was investigated. A three-strain cocktail of E. coli O157:H7 resistant to ampicillin was used to inoculate fresh and 48-h-old punctures, fresh-cut surfaces, and open or closed calyces. A concentric cutting procedure was used to evaluate depth of penetration within punctures and prevent cross contamination during sampling. Within 2 h, E. coli O157:H7 penetrated vertically through the fresh punctures and 3.4 mm within the underlying parenchyma. After 48 h, E. coli O157: H7 cells penetrated up to 5.5 mm within the punctures and >2.6 mm horizontally away from fresh punctures. However, 48-h-old punctures did not permit penetration beyond their boundaries. Fresh-cut surfaces permitted up to 2.8 mm penetration after 24 h. Onset of growth of E. coli O157:H7 occurred 4 to 8 h postinoculation on fresh punctures and fresh-cut surfaces with populations increasing by 3 logs after 48 h. E. coli O157:H7 penetrated within calyces regardless of the extent of opening or method of inoculation. However, E. coli O157:H7 was never recovered from the inner core of apples. Computed tomography scan imaging revealed that closed calyces effectively prevented penetration of sodium iodide solutions within the calyx cavity. Lack of solution penetration may explain why sanitizing treatments are ineffective in inactivating microbial cells within the calyx. Understanding the role of morphological differences in permitting or restricting bacterial penetration may lead to development of more effective strategies to enhance the safety of fresh horticultural products.     

Inactivation of Salmonella typhimurium and Escherichia coli O157:H7 in apple juice by a combination of nisin and cinnamon

Pasteurized apple juice with nisin (0, 25, 50, 100, and 200 ppm, wt/vol) and cinnamon (0 and 0.3%, wt/vol) was inoculated with Salmonella Typhimurium and Escherichia coli O157:H7 at 10(4) CFU/ml and stored at 5 and 20 degrees C. Counts on tryptic soy agar (TSA), selective medium (xylose Lysine desoxycholate agar for Salmonella Typhimurium, and MacConkey sorbitol agar for E. coli O157:H7), and thin agar layer (TAL) were determined at 1 h and 1, 3, 7, and 14 days. The TAL method (selective medium overlaid with TSA) was used for recovery of sublethally injured cells. The pathogens were gradually inactivated by the acidic pH of apple juice. Nisin and cinnamon greatly contributed to the inactivation. The killing effect was more marked at 20 degrees C, with counts in all treated samples being undetectable by direct plating in 3 days for Salmonella Typhimurium and 7 days for E. coli O157:H7. Thus, several factors influenced the decrease in counts: low pH, addition of nisin and cinnamon, and storage temperature. The TAL method was as effective as TSA in recovering injured cells of the pathogens. The combination of nisin and cinnamon accelerates death of Salmonella Typhimurium and E. coli O157:H7 in apple juice and so enhances the safety of the product.     

Comparison of recovery methods for freeze-injured Listeria monocytogenes,Salmonella Typhimurium,and Campylobacter coli in cell suspensions and associated with pork surfaces

Cells injured as a result of freezing, heating, and acidification treatments may not grow during conventional microbiological procedures owing to the presence of selective agents, compounds, or dyes in the media, impairing the cell's ability to repair itself and grow. Injured cells can be recovered by combining selective and nonselective media into a single system. With such combinations, the diffusion of the selective compounds or dyes is controlled, allowing for the resuscitation of injured cells of interest while also inhibiting the growth of undesirable background microflora. In this study, Listeria monocytogenes, Salmonella Typhimurium, and Campylobacter coli suspended in buffer or associated with pork surfaces were subjected to a freeze-thaw cycle (-15 degrees C for 24 h, 4 degrees C for 4 h). Following treatments, freeze-injured cells were plated on appropriate media for the overlay (OV), thin agar layer (TAL), and Lutri plate (LP) recovery methods. The levels of L. monocytogenes and Salmonella Typhimurium recovered from cell suspensions and pork surfaces by the TAL, OV, and LP methods following freeze treatments were not statistically different (P > 0.05) from recovery levels associated with nonselective media. Conversely, levels of pathogens on selective media were significantly reduced compared with those for the other methods employed. The TAL method's recovery of C. coli was not significantly different from that achieved with the nonselective media. Overall, the results presented in this study demonstrate that the TAL method not only was easier to perform, but also allowed improved isolation of single colonies for further characterization. This study may provide researchers with better methods to determine the effectiveness of industry-employed chilling processes in reducing pathogenic bacteria associated with red meat surfaces.