RESUMO
Refrigerated pickles are characterized by crisp, crunchy texture, opaque flesh, and fresh flavor. Typically produced without a thermal process, microbial safety relies on preventive controls, brine composition, and sufficient hold time prior to consumption. We hypothesized that brief blanching of whole cucumbers prior to pickling could provide an additional hurdle for pathogenic microbes without negatively impacting finished product quality. Blanch treatments (15, 90, or 180 s) in 80°C water were conducted in duplicate on two lots of cucumbers prior to cutting into spears, acidifying, and storing at 4°C. Enumeration of total aerobes, lactic acid bacteria, and glucose-fermenting coliforms was conducted for fresh and blanched cucumber. Texture, color, cured appearance development, and volatile compound profiles were analyzed for fresh and blanched cucumber and corresponding pickle products during refrigerated storage. The 90 s blanch consistently achieved a minimum 2-log reduction in cucumber microbiota and a predicted 5-log reduction of Escherichia coli O157:H7 up to 1.1 mm into the cucumber fruit. Blanching had no impact on tissue firmness during refrigerated storage for 1 year (p > 0.098). There were no differences in flavor-active lipid oxidation products (E,Z)-2,6-nonadienal and (E)-2-nonenal, and consumers (n = 110) were unable to differentiate between control and 90 s blanched cucumber pickles stored for 62 days. Exocarp color and mesocarp opacity were preserved by the blanching treatment, potentially extending product shelf life. This method offers processors an option for reducing the risk of microbial contamination while maintaining the quality attributes associated with refrigerated cucumber pickles. PRACTICAL APPLICATION: Refrigerated pickles do not undergo thermal processing, which can leave them vulnerable to microbial contamination. This study illustrates that adding a brief blanching step in refrigerated pickle processing can reduce indigenous microbiota without negatively impacting quality attributes. This blanching process could assist pickled vegetable manufacturers in providing additional safeguards for consumers while maintaining a high-quality product.
Assuntos
Cucumis sativus , Escherichia coli O157 , Alimentos Fermentados , Contagem de Colônia Microbiana , Cucumis sativus/microbiologia , FrutasRESUMO
Microbiological safety has been a critical issue for acid and acidified foods since it became clear that acid-tolerant pathogens such as Escherichia coli O157:H7 can survive (even though they are unable to grow) in a pH range of 3 to 4, which is typical for these classes of food products. The primary antimicrobial compounds in these products are acetic acid and NaCl, which can alter the intracellular physiology of E. coli O157:H7, leading to cell death. For combinations of acetic acid and NaCl at pH 3.2 (a pH value typical for non-heat-processed acidified vegetables), survival curves were described by using a Weibull model. The data revealed a protective effect of NaCl concentration on cell survival for selected acetic acid concentrations. The intracellular pH of an E. coli O157:H7 strain exposed to acetic acid concentrations of up to 40 mM and NaCl concentrations between 2 and 4% was determined. A reduction in the intracellular pH was observed for increasing acetic acid concentrations with an external pH of 3.2. Comparing intracellular pH with Weibull model predictions showed that decreases in intracellular pH were significantly correlated with the corresponding times required to achieve a 5-log reduction in the number of bacteria.
Assuntos
Ácido Acético/farmacologia , Escherichia coli O157/efeitos dos fármacos , Escherichia coli O157/crescimento & desenvolvimento , Modelos Biológicos , Cloreto de Sódio/farmacologia , Animais , Bovinos , Contagem de Colônia Microbiana , Escherichia coli O157/metabolismo , Microbiologia de Alimentos , Concentração de Íons de Hidrogênio , Verduras/microbiologiaRESUMO
ZnO/Nylon 6 nanofiber mats were prepared by an electrospinning-electrospraying hybrid process in which ZnO nanoparticles were dispersed on the surface of Nylon 6 nanofibers without becoming completely embedded. The prepared ZnO/Nylon 6 nanofiber mats were evaluated for their abilities to kill bacteria or inhibit their growth and to catalytically detoxify chemicals. Results showed that these ZnO/Nylon 6 nanofiber mats had excellent antibacterial efficiency (99.99%) against both the Gram-negative Escherichia coli and Gram-positive Bacillus cereus bacteria. In addition, they exhibited good detoxifying efficiency (95%) against paraoxon, a simulant of highly toxic chemicals. ZnO/Nylon 6 nanofiber mats were also deposited onto nylon/cotton woven fabrics and the nanofiber mats did not significantly affect the moisture vapor transmission rates and air permeability values of the fabrics. Therefore, ZnO/Nylon 6 nanofiber mats prepared by the electrospinning-electrospraying hybrid process are promising material candidates for protective applications.
RESUMO
Biofilm formation by Listeria monocytogenes is generally associated with its persistence in the food-processing environment. Serotype 1/2a strains make up more than 50% of the total isolates recovered from food and the environment, while serotype 4b strains are most often associated with major outbreaks of human listeriosis. Using a microplate assay with crystal violet staining, we examined biofilm formation by 18 strains of each serotype in tryptic soy broth with varying concentrations of glucose (from 0.25% to 10.0%, wt/vol), sodium chloride (from 0.5% to 7.0%, wt/vol) and ethanol (from 1% to 5.0%, vol/vol), and at different temperatures (22.5 degrees C, 30 degrees C, and 37 degrees C). A synergistic effect on biofilm formation was observed for glucose, sodium chloride, and temperature. The serotype 1/2a strains generally formed higher-density biofilms than the 4b strains under most conditions tested. Interestingly, most serotype 4b strains had a higher growth rate than the 1/2a strains, suggesting that the growth rate may not be directly related to the capacity for biofilm formation. Crystal violet was found to stain both bacterial cells and biofilm matrix material. The enhancement in biofilm formation by environmental factors was apparently due to the production of extracellular polymeric substances instead of the accumulation of viable biofilm cells.
Assuntos
Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Glucose/metabolismo , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/fisiologia , Cloreto de Sódio/metabolismo , Corantes/farmacologia , Etanol/metabolismo , Matriz Extracelular/metabolismo , Violeta Genciana/farmacologia , Humanos , Coloração e Rotulagem , TemperaturaRESUMO
The majority of Listeria monocytogenes isolates recovered from foods and the environment are strains of serogroup 1/2, especially serotypes 1/2a and 1/2b. However, serotype 4b strains cause the majority of human listeriosis outbreaks. Our investigation of L. monocytogenes biofilms used a simulated food-processing system that consisted of repeated cycles of growth, sanitation treatment, and starvation to determine the competitive fitness of strains of serotypes 1/2a and 4b in pure and mixed-culture biofilms. Selective enumeration of strains of a certain serotype in mixed-culture biofilms on stainless steel coupons was accomplished by using serotype-specific quantitative PCR and propidium monoazide treatment to prevent amplification of extracellular DNA or DNA from dead cells. The results showed that the serotype 1/2a strains tested were generally more efficient at forming biofilms and predominated in the mixed-culture biofilms. The growth and survival of strains of one serotype were not inhibited by strains of the other serotype in mixed-culture biofilms. However, we found that a cocktail of serotype 4b strains survived and grew significantly better in mixed-culture biofilms containing a specific strain of serotype 1/2a (strain SK1387), with final cell densities averaging 0.5 log(10) CFU/cm(2) higher than without the serotype 1/2a strain. The methodology used in this study contributed to our understanding of how environmental stresses and microbial competition influence the survival and growth of L. monocytogenes in pure and mixed-culture biofilms.
Assuntos
Antibiose , Biofilmes/crescimento & desenvolvimento , Microbiologia Ambiental , Listeria monocytogenes/fisiologia , Contagem de Colônia Microbiana/métodos , Humanos , Listeria monocytogenes/crescimento & desenvolvimento , Viabilidade Microbiana , Reação em Cadeia da Polimerase/métodos , Aço InoxidávelRESUMO
A number of studies on the influence of acid on Escherichia coli O157:H7 have shown considerable strain differences, but limited information has been reported to compare the acid resistance based on the different sources of E. coli O157:H7 isolates. The purpose of this study was to determine the survival of E. coli O157:H7 strains isolated from five sources (foods, bovine carcasses, bovine feces, water, and human) in 400 mM acetic acid solutions under conditions that are typical of acidified foods. The isolates from bovine carcasses, feces, and water survived acetic acid treatment at pH 3.3 and 30 degrees C significantly (P < or = 0.05) better than did any food or human isolates. However, resistance to acetic acid significantly increased as temperature decreased to 15 degrees C for a given pH, with little (P > or = 0.05) difference among the different isolation sources. All groups of E. coli O157:H7 strains showed more than 1.8- to 4.5-log reduction at pH 3.3 and 30 degrees C after 25 min. Significantly reduced (less than 1-log reduction) lethality for all E. coli O157:H7 strain mixtures was observed when pH increased to 3.7 or 4.3, with little difference in acetic acid resistance among the groups. The addition of glutamate to the acetic acid solution or anaerobic incubation provided the best protection compared with the above conditions for all groups of isolates. These results suggest that temperature, pH, and atmospheric conditions are key factors in establishing strategies for improving the safety of acidified foods.
Assuntos
Ácido Acético/farmacologia , Farmacorresistência Bacteriana , Escherichia coli O157/efeitos dos fármacos , Temperatura , Animais , Bovinos/microbiologia , Contagem de Colônia Microbiana , Microbiologia Ambiental , Escherichia coli O157/crescimento & desenvolvimento , Fezes , Contaminação de Alimentos , Manipulação de Alimentos/métodos , Humanos , Concentração de Íons de Hidrogênio , Microbiologia da ÁguaRESUMO
HIGHLIGHTS: NaCl and CaCl2 concentrations affected LAB and STEC strains differently. Growth rates at 6% NaCl were reduced for STEC more than LAB in vegetable broth. Extent of growth was reduced for STEC versus LAB for most vegetable fermentations. Death rates were minimally affected by salt type or concentration with lactic acid. Correlations between salt and STEC die-off were inconsistent for fermentation.
Assuntos
Escherichia coli O157 , Cloreto de Cálcio , Fermentação , Microbiologia de Alimentos , Concentração de Íons de Hidrogênio , Cloreto de Sódio , VerdurasRESUMO
Shiga toxin producing Escherichia coli (STEC) strains vary in acid resistance; however, little is known about the underlying mechanisms that result in strain specific differences. Among 25 STEC O157:H7 strains tested, 7 strains flocculated when grown statically for 18 h in minimal salts medium at 37°C, while 18 strains did not. Interestingly, the flocculation phenotype (cells came out of suspension) was found to correlate with degree of acid sensitivity in an assay with 400 mM acetic acid solution at pH 3.3 targeting acidified foods. Strains exhibiting flocculation were more acid sensitive and were designated FAS, for flocculation acid sensitive, while the acid resistant strain designated PAR for planktonic acid resistant. Flocculation was not observed for any strains during growth in complex medium (Luria Bertani broth). STEC strains B201 and B241 were chosen as representative FAS (2.4 log reduction) and PAR (0.15 log reduction) strains, respectively, due to differences in acid resistance and flocculation phenotype. Results from electron microscopy showed evidence of fimbriae production in B201, whereas fimbriae were not observed in B241.Curli fimbriae production was identified through plating on Congo red differential medium, and all FAS strains showed curli fimbriae production. Surprisingly, 5 PAR strains also had evidence of curli production. Transcriptomic and targeted gene expression data for B201 and B241indicated that csg and hde (curli and acid induced chaperone genes, respectively) expression positively correlated with the phenotypic differences observed for these strains. These data suggest that FAS strains grown in minimal medium express curli, resulting in a flocculation phenotype. This may be regulated by GcvB, which positively regulates curli fimbriae production and represses acid chaperone proteins. RpoS and other regulatory mechanisms may impact curli fimbriae production, as well. These findings may help elucidate mechanisms underlying differences among STEC strains in relating acid resistance and biofilm formation.
RESUMO
In many biological science and food processing applications, it is very important to control or modify pH. However, the complex, unknown composition of biological media and foods often limits the utility of purely theoretical approaches to modeling pH and calculating the distributions of ionizable species. This paper provides general formulas and efficient algorithms for predicting the pH, titration, ionic species concentrations, buffer capacity, and ionic strength of buffer solutions containing both defined and undefined components. A flexible, semi-mechanistic, partial buffering (SMPB) approach is presented that uses local polynomial regression to model the buffering influence of complex or undefined components in a solution, while identified components of known concentration are modeled using expressions based on extensions of the standard acid-base theory. The SMPB method is implemented in a freeware package, (pH)Tools, for use with Matlab. We validated the predictive accuracy of these methods by using strong acid titrations of cucumber slurries to predict the amount of a weak acid required to adjust pH to selected target values.
Assuntos
Íons/química , Modelos Químicos , Soluções/química , Algoritmos , Soluções Tampão , Cucumis sativus/química , Concentração de Íons de Hidrogênio , Concentração Osmolar , Sensibilidade e EspecificidadeRESUMO
The level of acid resistance among Escherichia coli O157:H7 strains varies, and strains with higher resistance to acid may have a lower infectious dose. The complete genome sequences belonging to two strains of Escherichia coli O157:H7 with different levels of acid resistance are presented here.
RESUMO
The objective of this study was to compare the acid resistance (AR) of non-O157 Shiga toxin-producing Escherichia coli (STEC) strains belonging to serogroups O26, O45, O103, O104, O111, O121, and O145 with O157:H7 STEC isolated from various sources in 400 mM acetic acid solutions (AAS) at pH 3.2 and 30 °C for 25 min with or without glutamic acid. Furthermore, the molecular subgrouping of the STEC strains was analyzed with the repetitive sequence-based PCR (rep-PCR) method using a DiversiLab(TM) system. Results for a total of 52 strains ranged from 0.31 to 5.45 log reduction CFU/mL in the absence of glutamic acid and 0.02 to 0.33 CFU/mL in the presence of glutamic acid except for B447 (O26:H11), B452 (O45:H2), and B466 (O104:H4) strains. Strains belonging to serogroups O111, O121, and O103 showed higher AR than serotype O157:H7 strains in the absence of glutamic acid. All STEC O157:H7 strains exhibited a comparable DNA pattern with more than 95% similarity in the rep-PCR results, as did the strains belonging to serogroups O111 and O121. Surprisingly, the DNA pattern of B458 (O103:H2) was similar to that of O157:H7 strains with 82% similarity, and strain B458 strain showed the highest AR to AAS among the O103 strains with 0.44 log reduction CFU/mL without glutamic acid. In conclusion, STEC serotypes isolated from different sources exhibited diverse AR and genetic subtyping patterns. Results indicated that some non-O157 STEC strains may have higher AR than STEC O157:H7 strains under specific acidic conditions, and the addition of glutamic acid provided enhanced protection against exposure to AAS.
Assuntos
Ácido Acético/farmacologia , DNA Bacteriano/análise , Escherichia coli O157/genética , Sorogrupo , Toxinas Shiga , Escherichia coli Shiga Toxigênica/genética , Escherichia coli O157/efeitos dos fármacos , Escherichia coli O157/metabolismo , Proteínas de Escherichia coli , Reação em Cadeia da Polimerase , Escherichia coli Shiga Toxigênica/efeitos dos fármacos , Escherichia coli Shiga Toxigênica/metabolismoRESUMO
Bacillus licheniformis has been shown to cause pH elevation in tomato products having an initial pH below 4.6 and metabiotic effects that can lead to the growth of pathogenic bacteria. Because of this, the organism poses a potential risk to acidified vegetable products; however, little is known about the growth and metabolism of this organism in these products. To clarify the mechanisms of pH change and growth of B. licheniformis in vegetable broth under acidic conditions, a cucumber juice medium representative of a noninhibitory vegetable broth was used to monitor changes in pH, cell growth, and catabolism of sugars and amino acids. For initial pH values between pH 4.1 to 6.0, pH changes resulted from both fermentation of sugar (lowering pH) and ammonia production (raising pH). An initial pH elevation occurred, with starting pH values of pH 4.1 to 4.9 under both aerobic and anaerobic conditions, and was apparently mediated by the arginine deiminase reaction of B. licheniformis. This initial pH elevation was prevented if 5 mM or greater acetic acid was present in the brine at the same pH. In laboratory media, under favorable conditions for growth, data indicated that growth of the organism was inhibited at pH 4.6 with protonated acetic acid concentrations of 10 to 20 mM, corresponding to 25 to 50 mM total acetic acid; however, growth inhibition required greater than 300 mM citric acid (10-fold excess of the amount in processed tomato products) products under similar conditions. The data indicate that growth and pH increase by B. licheniformis may be inhibited by the acetic acid present in most commercial acidified vegetable products but not by the citric acid in many tomato products.
Assuntos
Ácido Acético/farmacologia , Arginina/farmacologia , Bacillus/efeitos dos fármacos , Cucumis sativus/microbiologia , Sucos de Frutas e Vegetais/microbiologia , Bacillus/crescimento & desenvolvimento , Ácido Cítrico/farmacologia , Contagem de Colônia Microbiana , Fermentação , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Concentração de Íons de Hidrogênio , Sais/químicaRESUMO
The biochemistry and microbial ecology of 2 similar types of watery (mul) kimchi, containing sliced and unsliced radish and vegetables (nabak and dongchimi, respectively), were investigated. Samples from kimchi were fermented at 4, 10, and 20 °C were analyzed by plating on differential and selective media, high-performance liquid chromatography, and high-throughput DNA sequencing of 16S rDNA. Nabak kimchi showed similar trends as dongchimi, with increasing lactic and acetic acids and decreasing pH for each temperature, but differences in microbiota were apparent. Interestingly, bacteria from the Proteobacterium phylum, including Enterobacteriaceae, decreased more rapidly during fermentation at 4 °C in nabak cabbage fermentations compared with dongchimi. Although changes for Proteobacterium and Enterobacteriaceae populations were similar during fermentation at 10 and 20 °C, the homolactic stage of fermentation did not develop for the 4 and 10 °C samples of both nabak and dongchimi during the experiment. These data show the differences in biochemistry and microbial ecology that can result from preparation method and fermentation conditions of the kimchi, which may impact safety (Enterobacteriaceae populations may include pathogenic bacteria) and quality (homolactic fermentation can be undesirable, if too much acid is produced) of the product. In addition, the data also illustrate the need for improved methods for identifying and differentiating closely related lactic acid bacteria species using high-throughput sequencing methods.
Assuntos
DNA Bacteriano/isolamento & purificação , Fermentação , Microbiologia de Alimentos , Verduras/microbiologia , DNA Bacteriano/química , DNA Bacteriano/classificação , DNA Ribossômico/química , Enterobacteriaceae/classificação , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Lactobacillus/classificação , Proteobactérias/classificação , Proteobactérias/genética , Proteobactérias/isolamento & purificação , RNA Ribossômico 16S/genética , Raphanus/microbiologia , Análise de Sequência de DNA , TemperaturaRESUMO
In this study, the adhesion of bacteria to fresh cucumber surfaces in aqueous suspension was shown to be dependent on time of incubation, inoculum species and concentration, and temperature. The adhesion of bacteria to the fruit in wash water was less extensive at lower temperatures and shorter exposure times. Various species of bacteria were adsorbed to cucumber surfaces in the following relative order: Salmonella Typhimurium > Staphylococcus aureus > Lactobacillus plantarum > Listeria monocytogenes. Cells were adsorbed at all temperatures tested (5, 15, 25, and 35 degrees C) at levels that depended on incubation time, but the numbers of cells adsorbed were larger at higher incubation temperatures. Levels of adhesion of bacteria to dewaxed fruit were higher for L. monocytogenes and lower for Salmonella Typhimurium, L. plantarum, and S. aureus than were levels of adhesion to waxed fruit.
Assuntos
Cucumis sativus/microbiologia , Contaminação de Alimentos , Microbiologia de Alimentos , Aderência Bacteriana , Cucumis sativus/ultraestrutura , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/fisiologia , Listeria monocytogenes/crescimento & desenvolvimento , Listeria monocytogenes/fisiologia , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/fisiologia , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/fisiologia , Temperatura , Fatores de TempoRESUMO
UNLABELLED: Refrigerated cucumber pickle products cannot be heat processed due to the loss of characteristic sensory attributes. Typically brined refrigerated pickles contain less than 100 mM acetic acid with pH values of 3.7 to 4.0. Refrigeration (4 to 10 °C) helps to inhibit the growth of spoilage bacteria and maintain flavor, texture, and appearance of the pickles. Previous research has shown that pathogenic Escherichia coli strains are unusually acid resistant and survive better in refrigerated acid solutions than at higher temperatures. We found that E. coli O157:H7 can survive for 1 mo or longer at 4 °C in brines typical of commercial refrigerated pickles. Our objective was to develop methods to assure a 5-log reduction of pathogenic E. coli in these types of products, while maintaining the sensory characteristics. A novel brine formulation was developed, based on current commercial refrigerated pickle brines, which contained 25 mM fumaric acid, 5 mM benzoic acid, 70 mM acetic acid, and 342 mM (2%) sodium chloride, with a pH of 3.8. Sensory data indicate that this formulation did not affect flavor or other sensory attributes of the product, compared to traditional formulations. We achieved a 5-log reduction of E. coli O157:H7 at 30 °C for 1.52 ± 0.15 d, at 20 °C for 3.12 ± 0.34 d, or at 10 °C for 8.83 ± 0.56 d. Growth of lactic acid bacteria was also inhibited. These results can be used by manufacturers to assure a 5-log reduction in cell numbers of E. coli O157:H7 and Salmonella without a heat process during the manufacture of refrigerated pickle products. PRACTICAL APPLICATION: While refrigerated acidified vegetable products are exempt from the acidified foods regulations, we have shown that the vegetative microbial pathogens E. coli O157:H7 can survive for up to 1 mo in these products, given current commercial production practices. To improve the safety of refrigerated pickle products, a brine formulation with reduced acetic acid, but containing fumaric acid, was developed to assure a 5-log reduction in cell numbers of E. coli O157:H7 without a heat process. The formulation can be used to assure the safety of refrigerated pickled vegetables without altering sensory characteristics.
Assuntos
Cucumis sativus/microbiologia , Escherichia coli O157/crescimento & desenvolvimento , Contaminação de Alimentos/prevenção & controle , Manipulação de Alimentos/métodos , Microbiologia de Alimentos , Refrigeração/métodos , Ácido Acético/análise , Contagem de Colônia Microbiana , Comportamento do Consumidor , Qualidade de Produtos para o Consumidor , Cucumis sativus/química , Fermentação , Conservação de Alimentos/métodos , Fumaratos/análise , Humanos , Concentração de Íons de Hidrogênio , Ácido Láctico/análise , Dinâmica não Linear , Salmonella/crescimento & desenvolvimento , Sais/análise , Sais/químicaRESUMO
UNLABELLED: Bacterial pathogens have been reported on fresh cucumbers and other vegetables used for commercial fermentation. The Food and Drug Administration currently has a 5-log reduction standard for E. coli O157:H7 and other vegetative pathogens in acidified pickle products. For fermented vegetables, which are acid foods, there is little data documenting the conditions needed to kill acid resistant pathogens. To address this knowledge gap, we obtained 10 different cucumber fermentation brines at different stages of fermentation from 5 domestic commercial plants. Cucumber brines were used to represent vegetable fermentations because cabbage and other vegetables may have inhibitory compounds that may affect survival. The 5-log reduction times for E. coli O157:H7 strains in the commercial brines were found to be positively correlated with brine pH, and ranged from 3 to 24 d for pH values of 3.2 to 4.6, respectively. In a laboratory cucumber juice medium that had been previously fermented with Lactobacillus plantarum or Leuconostoc mesenteroides (pH 3.9), a 5-log reduction was achieved within 1 to 16 d depending on pH, acid concentration, and temperature. During competitive growth at 30 °C in the presence of L. plantarum or L. mesenteroides in cucumber juice, E. coli O157:H7 cell numbers were reduced to below the level of detection within 2 to 3 d. These data may be used to aid manufacturers of fermented vegetable products determine safe production practices based on fermentation pH and temperature. PRACTICAL APPLICATION: Disease causing strains of the bacterium E. coli may be present on fresh vegetables. Our investigation determined the time needed to kill E. coli in cucumber fermentation brines and how E. coli strains are killed in competition with naturally present lactic acid bacteria. Our results showed how brine pH and other brine conditions affected the killing of E. coli strains. These data can be used by producers of fermented vegetable products to help assure consumer safety.
Assuntos
Cucumis sativus/microbiologia , Escherichia coli O157/crescimento & desenvolvimento , Conservação de Alimentos/métodos , Frutas/microbiologia , Viabilidade Microbiana , Ácido Acético/metabolismo , Antibiose , Contagem de Colônia Microbiana , Escherichia coli O157/metabolismo , Fermentação , Frutose/metabolismo , Glucose/metabolismo , Concentração de Íons de Hidrogênio , Ácido Láctico/metabolismo , Lactobacillus plantarum/crescimento & desenvolvimento , Lactobacillus plantarum/metabolismo , Leuconostoc/crescimento & desenvolvimento , Leuconostoc/metabolismo , Concentração Osmolar , Sais/química , Cloreto de Sódio na Dieta/análise , Temperatura , Fatores de TempoRESUMO
Outbreaks of disease due to vegetative bacterial pathogens associated with acid foods (such as apple cider) have raised concerns about acidified vegetables and related products that have a similar pH (3.2 to 4.0). Escherichia coli O157:H7 and related strains of enterohemorrhagic E. coli (EHEC) have been identified as the most acid resistant vegetative pathogens in these products. Previous research has shown that the lack of dissolved oxygen in many hermetically sealed acid or acidified food products can enhance survival of EHEC compared with their survival under aerobic conditions. We compared the antimicrobial effects of several food acids (acetic, malic, lactic, fumaric, benzoic, and sorbic acids and sulfite) on a cocktail of EHEC strains under conditions representative of non-heat-processed acidified vegetables in hermetically sealed jars, holding the pH (3.2) and ionic strength (0.342) constant under anaerobic conditions. The overall antimicrobial effectiveness of weak acids used in this study was ranked, from most effective to least effective: sulfite > benzoic acid > sorbic acid > fumaric acid > L- and D-lactic acid > acetic acid > malic acid. These rankings were based on the estimated protonated concentrations required to achieve a 5-log reduction in EHEC after 24 h of incubation at 30°C. This study provides information that can be used to formulate safer acid and acidified food products and provides insights about the mode of action of weak acids against EHEC.
Assuntos
Ácidos/farmacologia , Antibacterianos/farmacologia , Escherichia coli O157/efeitos dos fármacos , Contaminação de Alimentos/análise , Conservação de Alimentos/métodos , Oxigênio/farmacologia , Contagem de Colônia Microbiana , Qualidade de Produtos para o Consumidor , Escherichia coli O157/crescimento & desenvolvimento , Humanos , Concentração de Íons de Hidrogênio , Concentração OsmolarRESUMO
Six bacteriophages active against Leuconostoc fallax strains were isolated from industrial sauerkraut fermentation brines. These phages were characterized as to host range, morphology, structural proteins, and genome fingerprint. They were exclusively lytic against the species L. fallax and had different host ranges among the strains of this species tested. Morphologically, three of the phages were assigned to the family Siphoviridae, and the three others were assigned to the family Myovidae: Major capsid proteins detected by electrophoresis were distinct for each of the two morphotypes. Restriction fragment length polymorphism analysis and randomly amplified polymorphic DNA fingerprinting showed that all six phages were genetically distinct. These results revealed for the first time the existence of bacteriophages that are active against L. fallax and confirmed the presence and diversity of bacteriophages in a sauerkraut fermentation. Since a variety of L. fallax strains have been shown to be present in sauerkraut fermentation, bacteriophages active against L. fallax are likely to contribute to the microbial ecology of sauerkraut fermentation and could be responsible for some of the variability observed in this type of fermentation.
Assuntos
Bacteriófagos/isolamento & purificação , Genoma Viral , Leuconostoc/virologia , Bacteriófagos/genética , Bacteriófagos/crescimento & desenvolvimento , Bacteriófagos/metabolismo , Bacteriófagos/ultraestrutura , Impressões Digitais de DNA , DNA Viral/análise , Fermentação , Polimorfismo de Fragmento de Restrição , Técnica de Amplificação ao Acaso de DNA Polimórfico , Proteínas Virais/análiseRESUMO
We developed a mechanistic mathematical model for predicting the progression of batch fermentation of cucumber juice by Lactococcus lactis under variable environmental conditions. In order to overcome the deficiencies of presently available models, we use a dynamic energy budget approach to model the dependence of growth on present as well as past environmental conditions. When parameter estimates from independent experimental data are used, our model is able to predict the outcomes of three different temperature shift scenarios. Sensitivity analyses elucidate how temperature affects the metabolism and growth of cells through all four stages of fermentation and reveal that there is a qualitative reversal in the factors limiting growth between low and high temperatures. Our model has an applied use as a predictive tool in batch culture growth. It has the added advantage of being able to suggest plausible and testable mechanistic assumptions about the interplay between cellular energetics and the modes of inhibition by temperature and end product accumulation.
Assuntos
Metabolismo Energético , Lactococcus lactis/metabolismo , Modelos Biológicos , Calibragem , Fermentação , Reprodutibilidade dos Testes , TemperaturaRESUMO
Lactic acid bacterial strains were isolated from brines sampled after 7 days of an industrial sauerkraut fermentation, and six strains were selected on the basis of susceptibility to bacteriophages. Bacterial growth in cabbage juice was monitored, and the fermentation end products were identified, quantified, and compared to those of Leuconostoc mesenteroides. Identification by biochemical fingerprinting, endonuclease digestion of the 16S-23S intergenic transcribed spacer region, and sequencing of variable regions V1 and V2 of the 16S rRNA gene indicated that the six selected sauerkraut isolates were Leuconostoc fallax strains. Random amplification of polymorphic DNA fingerprints indicated that the strains were distinct from one another. The growth and fermentation patterns of the L. fallax isolates were highly similar to those of L. mesenteroides. The final pH of cabbage juice fermentation was 3.6, and the main fermentation end products were lactic acid, acetic acid, and mannitol for both species. However, none of the L. fallax strains exhibited the malolactic reaction, which is characteristic of most L. mesenteroides strains. These results indicated that in addition to L. mesenteroides, a variety of L. fallax strains may be present in the heterofermentative stage of sauerkraut fermentation. The microbial ecology of sauerkraut fermentation appears to be more complex than previously indicated, and the prevalence and roles of L. fallax require further investigation.