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Detection, identification and characterization of bacteriocin-producing lactic acid bacteria from retail food products.

Forty bacteriocin-producing (Bac+) lactic acid bacteria (LAB) were isolated from food samples purchased from retail supermarkets and local farms. Of the 40 Bac+ isolates, 18 were isolated from 85 food samples by enrichment (21% isolation rate) whereas eight were obtained from 63 samples by direct plating (13% isolation rate). By direct plating, Bac+ LAB were detected at levels up to 2.4 x 10(5) cfu/g in ready-to-eat meats. The Bac+ isolates were identified by carbohydrate fermentation patterns, SDS-PAGE protein patterns, and other biochemical characteristics; SDS-PAGE proved invaluable in identifying strains that could not be identified by other means. Differential inhibitory spectra against indicator microorganisms assisted in the identification of 19 unique Bac+ isolates. Bac+ LAB included Enterococcus faecalis, Lactobacillus curvatus, Lb. delbrueckii, Lb. plantarum, Lactococcus lactis, and Pediococcus acidilactici. Lb. curvatus (four strains) and Lc. lactis (nine strains) were the only isolates inhibitory to foodborne pathogens including Listeria monocytogenes, Bacillus cereus, Clostridium perfringens and Staphylococcus aureus. Some Lc. lactis isolates inhibited as many as nine Gram-positive genera. Lb. curvatus FS47 and FS65 grew to high cell densities and produced bacteriocin at 6 degrees C; however, Lc. lactis FS56 produced greater levels of bacteriocin at lower cell densities. The high incidence of Bac+ LAB detected in retail foods indicates that the public is consuming a wide variety of Bac+ LAB that occur as natural contaminants. These data suggest a greater role for bacteriocins as biopreservatives in food.

Animals↗

Antibacterial activity of Lactobacillus plantarum UG1 isolated from dry sausage: characterization, production and bactericidal action of plantaricin UG1.

Lactobacillus plantarum UG1 isolated from dry sausage produced an antimicrobial substance that inhibited other strains of the genera Lactobacillus and Lactococcus, and some foodborne pathogens including Listeria monocytogenes, Bacillus cereus, Clostridium perfringens and Clostridium sporogenes. This antibacterial substance was inactivated by proteolytic enzymes and showed a bactericidal mode of action. Consequently, it was characterized as a bacteriocin, and was designated plantaricin UG1. This bacteriocin was stable in the pH range 4.5 to 7.0, partially inactivated by amylolytic enzymes and relatively thermostable. It was not affected by organic or lipolytic enzymes. Production of plantaricin UG1 was pH- and temperature-dependant and maximum yields were obtained in MRS broth cultures maintained at initial pH 6.5, and incubated at 25 degrees C to 30 degrees C, in the exponential to the early stationary growth phase of the producer organism. Ultrafiltration studies indicated that plantaricin UG1 has a molecular weight between 3 and 10 KDa. Curing experiments with L. plantarum UG1 resulted in the appearance of variants that lost bacteriocin production ability but were still immune to the bacteriocin. Plantaricin UG1 production appeared to be chromosomal encoded. Sensitive and insensitive Gram-positive bacteria adsorbed plantaricin UG1 irrespective of their susceptibility to it. In contrast, Gram-negative bacteria did not adsorb plantaricin UG1. The bactericidal action of plantaricin UG1 did not depend on the physiological state of the indicator culture and did not cause cell lysis. The resistance of two indicator strains to plantaricin UG1 has been studied.

Adsorption↗

Rumen contents as a reservoir of enterohemorrhagic Escherichia coli.

We investigated the role of the rumen fermentation as a barrier to the foodborne pathogen, Escherichia coli O157:H7. Strains of E. coli, including several isolates of O157:H7, grew poorly in media which simulated the ruminal environment of a well-fed animal. Strains of E. coli O157:H7 did not display a superior tolerance to ruminal conditions which may facilitate their colonization of the bovine digestive tract. Unrestricted growth of E. coli was observed in rumen fluid collected from fasted cattle. Growth was inhibited by rumen fluid collected from well-fed animals. Well-fed animals appear less likely to become reservoirs for pathogenic E. coli. These results have implications for cattle slaughter practices and epidemiological studies of E. coli O157:H7.

Anaerobiosis↗

A molecular beacon DNA microarray system for rapid detection of E. coli O157:H7 that eliminates the risk of a false negative signal.

A DNA hybridization based optical detection platform for the detection of foodborne pathogens has been developed with virtually zero probability of the false negative signal. This portable, low-cost and real-time assaying detection platform utilizes the color changing molecular beacon as a probe for the optical detection of the target sequence. The computer-controlled detection platform exploits the target hybridization induced change of fluorescence color due to the Förster (fluorescence) resonance energy transfer (FRET) between a pair of spectrally shifted fluorophores conjugated to the opposite ends of a beacon (oligonucleotide probe). Unlike the traditional fluorophore-quencher beacon design, the presence of two fluorescence molecules allows to actively visualize both hybridized and unhybridized states of the beacon. This eliminates false negative signal detection characteristic for the fluorophore-quencher beacon where bleaching of the fluorophore or washout of a beacon is indistinguishable from the absence of the target DNA sequence. In perspective, the two-color design allows also to quantify the concentration of the target DNA in a sample down to < =1 ng/microl. The new design is suitable for simultaneous reliable detection of hundreds of DNA target sequences in one test run using a series of beacons immobilized on a single substrate in a spatial format.

DNA, Bacterial↗

Occurrence of non-O157 shiga toxin-producing Escherichia coli in ready-to-eat food from supermarkets in Argentina.

Between June 2000 and December 2001, 500 food samples were collected from supermarkets and shops selling ready-to-eat food in Rosario, Argentina, and examined for Escherichia coli. Forty-nine E. coli isolates from food samples were further characterized for virulence genes by multiplex polymerase chain reaction (PCR) targeting the stx1, stx2, stx2e, eaeA, CNF1, CNF2, Einv, LTI, STI, and STII genes in four groups. Out of 49 E. coli isolates screened by multiplex PCR, only 10 possessed Shiga toxin genes, stx1 and stx2 genes and none possessed the other genes. The Shiga toxin positive E. coli strains (STEC) were isolated from soft, cottage cheeses, chicken with sauce and vegetables mayonase. These E. coli isolates were serogrouped and belonged to O18 (two strains), O8, O57w, O79, O44, and O128; three strains were untypeable. Pulsed-field gel electrophoresis (PFGE) with XbaI generated a unique profile for each, having 10-15 bands ranging from 50 to 500 kb, except that strain ARG 20 generated small bands and was partly degraded. These strains are potential foodborne pathogens and their presence in ready-to-eat food illustrates the need to keep a careful watch for the source of pathogens and then develop methods to control them.

Argentina↗

Inhibition of Listeria monocytogenes by a lactic acid bacterium isolated from Italian salami.

Listeria monocytogenes is an opportunistic psychrotroph foodborne pathogen that has been used as a model organism to study the efficacy of many different preservation methods. This work aimed to test the antilisterial activity of lactic acid bacteria isolated from Italian salami and study the development of resistance. Isolates were obtained from naturally fermented Italian salami and cultures that retained activity in the supernatants after pH neutralization and catalase treatment were further characterized. The isolate showing highest inhibitory activity (PD 6.9) was tested for sensibility to proteases, heat and pH. To evaluate if resistance developed, sensitive strains were transferred with sub-lethal doses of the partially purified inhibitory substance and then inoculated into media containing higher doses of the extract. Isolate PD 6.9 inhibited several L. monocytogenes strains obtained from different origins and retained its activity over a wide range of pH and temperature. When increasing concentrations (10-100 AU ml(-1)) of the partially purified inhibitory substance were added to culture media, growth of L. monocytogenes did not occur even after 12 h of incubation. Cultures of Listeria that were transferred with sub-lethal doses (10 AU ml(-1)) of the partially purified inhibitory substance could resist higher doses of the extract (50 AU ml(-1)), but were inhibited when the concentration was further increased (100 AU ml(-1)). These results indicate that isolate PD 6.9 could potentially be used as a bioprotective culture for salami fermentation.

Animals↗

Metal-organic frameworks nanozyme-integrated portable microneedle patch for visual bacterial monitoring in meat.

Foodborne microbial contamination is a major global health concern, with conventional methods often being time-consuming and complex. Herein, we developed a novel portable biosensor by integrating microneedle patch technology and a metal-organic framework (Fe/Cu-NBDC MOF) nanozyme, enabling rapid, on-site, visual detection of bacteria in meat. The sensing system works by encapsulating aptamer-functionalized MOF nanozymes within a hydrogel patch, where their catalytic sites are initially blocked by the aptamer. In the presence of Staphylococcus aureus (S. aureus) as the target, the specific aptamer's binding to bacteria exposes numerous catalytic sites, further activating the chromogenic reaction of the tetramethylbenzidine&#x2011;hydrogen peroxide (TMB-H&#x2082;O&#x2082;) system, enabling visual detection of S. aureus. The biosensor demonstrates a detection limit of 82&#xa0;CFU/mL with excellent specificity to successfully apply to commercial mutton. By integrating sampling, enrichment, and visual detection into a single compact device, this platform offers a practical, efficient solution for rapid on-site screening of foodborne pathogens.

Biosensing Techniques↗

Artificial neural network data fusion-mediated dual-mode sensor based on Fe3O4@PdIr for Salmonellatyphimurium detection in food.

Salmonella Typhimurium (S. typhimurium) is a major foodborne pathogen that poses a serious threat to public health. In this study, a colorimetric/electrochemical dual-mode biosensor assisted by artificial neural network (ANN) was developed for the sensitive detection of S. typhimurium. Fe3O4@PdIr nanocomposites with enhanced peroxidase-like activity and electrochemical performance were prepared and conjugated with an aptamer specific to S. typhimurium to obtain Fe3O4@PdIr-Apt. Through the sandwich binding of Fe3O4@PdIr-Apt and Apt to the target, the nanocomposites were attached to microplates or Au electrodes, thereby generating colorimetric and electrochemical signals. The ANN model deeply resolved the complex nonlinear relationship between the dual signals, enabling mutual correction and ultimately performing data fusion to output a single detection result, which significantly reduced the mean square error while improving detection sensitivity and reliability. This sensor exhibited a wide linear range of 2.7-2.7&#xa0;&#xd7;&#xa0;108&#xa0;CFU/mL and a low detection limit of 1.66&#xa0;CFU/mL. Additionally, this method was successfully applied to the detection of S. typhimurium in pork and milk, with a recovery rate of 95.19%&#xa0;&#x223c;&#xa0;104.07%. It indicated that the constructed sensor holds great practical potential for S. typhimurium detection.

Neural Networks, Computer↗

Regional genomic analysis of lineage distribution and transferable multidrug resistance among chicken-associated Salmonella Kentucky isolates in China.

Salmonella enterica serovar Kentucky is an important multidrug-resistant foodborne pathogen in the poultry meat supply chain. Although recent broader genomic studies have elucidated the population structure and epidemiological significance of major lineages in China (e.g., ST198 and ST314), the regional dynamics within local poultry supply chains remain insufficiently characterized. In this study, 31 chicken meat-derived isolates from Shanghai and 39 publicly available genomes from China were analyzed using antimicrobial susceptibility testing, whole-genome sequencing, phylogenetic analysis, conjugation experiments, and complete sequencing of representative plasmids. This enabled a systematic characterization of the molecular epidemiological features of the population and the mechanisms underlying resistance dissemination. Population genomic analysis revealed a lineage composition markedly different from the global epidemiological pattern: ST314 was the predominant sequence type among the Shanghai chicken-derived isolates (74.2%), whereas the internationally recognized high-risk clone ST198 accounted for only 25.8% of the local isolates. However, risk stratification analysis indicated that although ST198 was detected less frequently, it carried a significantly greater burden of acquired resistance genes and therefore represented a higher-risk resistant lineage. Functional and structural validation further elucidated the molecular basis of resistance dissemination within this high-risk lineage. Conjugation experiments confirmed the co-transfer of a multidrug resistance module carrying blaTEM-1 and blaCTX-M-267 to the recipient strain Escherichia coli J53. Complete plasmid analysis revealed that these two &#x3b2;-lactam resistance genes were co-localized on a 242-kb transferable plasmid flanked by Tn1331, Tn3, and multiple transposase-associated elements, thereby providing a structural basis for their horizontal transfer. This study provides important molecular epidemiological evidence for lineage-specific surveillance and risk-stratified control of resistant Salmonella in the poultry meat supply chain and further underscores the need for continuous monitoring of mobile genetic elements within a One Health framework.

Animals↗

Inhibitory mechanism of phloretin on the AgrA LytTR domain-agr operon complex formation and its application in beef.

Staphylococcus aureus (S. aureus) represents a major foodborne pathogen whose enterotoxin production poses significant challenges to food safety due to its high environmental resistance and limited efficacy of conventional sterilization. Since the expression of enterotoxins is predominantly governed by the agr quorum sensing system, targeting this regulatory pathway has become a strategic choice for virulence control. This study elucidated the mechanism by which phloretin, a potential quorum sensing inhibitor, interferes with the agr system to attenuate virulence. To achieve this, the recombinant AgrA LytTR domain was expressed and purified, and its interaction with phloretin was characterized using thermal shift assays (TSA), electrophoretic mobility shift assays (EMSA), and molecular dynamics (MD) simulations. The results showed that phloretin specifically binds to the AgrA LytTR domain, enhancing its thermal stability and disrupting AgrA LytTR-agr operon binding by reducing the free energy of interaction between them, without causing significant structural rearrangement. Mechanistic analysis indicated that phloretin sterically hinders key &#x3b2;-sheet turn residues (HIS169, ASN201, ARG233), thereby impairing DNA recognition, downregulating RNAIII transcription, and inhibiting agr signaling. In cooked beef, phloretin significantly inhibited the secretion of enterotoxins and &#x3b1;-hemolysin, while delaying lipid oxidation and protein degradation, and maintaining the meat texture. These findings suggested that phloretin is a multifunctional substance with anti-virulence, antioxidant, and preservative properties, demonstrating its potential as a natural food preservative.

Phloretin↗

Temporal shifts in gyrA mutation types and sublineage replacement in ST11 Salmonella enterica&#xa0;serovar Enteritidis over a decade (2014-2023): A genomic epidemiological study in Guangxi, China.

The overuse or abuse of antibiotics drives the global health threat of antimicrobial resistance. Although bans on certain veterinary antibiotics, such as colistin, have proven effective, the impact of fluoroquinolone stewardship on the evolution of the foodborne pathogen Salmonella enterica serovar Enteritidis (S. Enteritidis) remains unclear. Here, we conducted a decade-long (2014-2023) retrospective longitudinal genomic epidemiological analysis of 441&#xa0;ST11 S. Enteritidis isolates from Guangxi, China, alongside a global reference dataset of 4297 genomes. Our aim was to elucidate the effect of real-world antibiotic stewardship on the shift of gyrA point mutations and lineage distribution. Surveillance identified three global epidemic clade sublineages (GEC-L2, L3, L4), with the multidrug-resistant GEC-L4 (i.e., GC-c or MMC2), characterized by the gyrA mutation with amino acid substitution D87Y, being domestically dominant (70.07%, 309/441). Following China's 2016 ban on the veterinary use of critical fluoroquinolones, the proportion of the highly resistant GEC-L4 sublineage decreased continuously (from 86.84% in 2017 to 56.00% in 2023), while the less resistant GEC-L3 sublineage (i.e., GC-b or MMC1), mainly characterized by gyrA D87G, increased simultaneously (from 13.16% to 44.00%). This phenomenon might be attributed to the fact that the GEC-L4 sublineage exhibited a higher fitness cost compared with the GEC-L3 sublineage, as confirmed by the competition assay. A Random Forest Model validated that the gyrA mutation with amino acid substitution&#xa0;D87Y was the paramount feature for these sublineages' identification. In contrast, global data showed a continuous increase in gyrA mutations (from 8.63% in 2006 to 68.85% in 2024), primarily D87Y (from 1.44% to 31.15%) and D87N (from 4.32% to 22.95%), correlating with rising average fluoroquinolone consumption. This study provides direct genomic evidence that national-level antibiotic stewardship can drive the replacement of highly resistant sublineages with moderately resistant ones. These findings offer crucial scientific evidence for evaluating the impact of antibiotic management policies and inform strategies for the rational use of antimicrobials.

China↗

Quantitative contamination and transfer of Escherichia coli from foods by houseflies, Musca domestica L. (Diptera: Muscidae).

The housefly, Musca domestica L. (Diptera: Muscidae), is recognized as an important factor in the dissemination of various infectious diseases such as cholera, shigellosis, and salmonellosis. They can also serve as a cross-contamination vector for other foodborne pathogens. However, the potential for bacterial transfer by houseflies has been demonstrated in a qualitative rather than quantitative manner. In this study, the numbers of bacteria a housefly can carry on its body and transfer to a clean surface after exposure to a sugar-milk aqueous solution, steak, and potato salad contaminated with a fluorescent gene Escherichia coli (8 log10 CFU/ml) were determined. In the first series of experiments to quantify bacterial numbers on the flies, about 40-60 flies were transferred into a sterile cage, exposed to the food for 30 min, the flies immobilized and the attached E. coli on each fly enumerated. Detectable E. coli (>1.7 log10 CFU/fly) were found on 43% (29/67), 53% (23/43), and 62% (32/52) of the flies in the cages with sugar/milk, steak, and potato salad, respectively. For the positive flies, the geometric mean carriage (log10 CFU/fly) was 2.93+/-1.24 for sugar-milk, 3.77+/-1.28 for steak, and 2.25+/-0.64 for the potato salad. In the second series of experiments, the transfer of bacteria by individual flies from contaminated food to the inner surface of a sterile jar per each landing was determined. E. coli transferred from the sugar-milk was 3.5+/-0.7 log10 CFU/fly-landing, 3.9+/-0.7 for steak and 2.61+/-1.16 for the potato salad. From the initial contamination levels of bacteria and the number of transferred bacteria, it can be calculated that flies contaminate clean surfaces with approximately 0.1 mg of food per landing.

Animals↗

Factors affecting production of extracellular carbohydrate complexes by Escherichia coli O157:H7.

Production of extracellular carbohydrate complexes (ECC) by foodborne pathogens on raw fruits and vegetables may result in protection against removal or inactivation by sanitizers. The influence of environmental conditions on cell growth, the total amount of ECC produced, and the amount of ECC produced on a per cell basis by Escherichia coli O157:H7 strains ATCC 43895 (wild type) and 43895-exopolysaccharides (EPS) (natural mutant, extensive EPS producer) was studied. To determine the effects of pH on the production of ECC on a per cell basis, E. coli O157:H7 was grown aerobically at 12 and 22 degrees C on tryptic soy agar (TSA) acidified to pH 7.0, 6.5, 6.0, 5.5, 5.0, 4.5, and 4.0. Lettuce, alfalfa sprout, cantaloupe, tomato, and apple juice agars (pH 4.46-6.50) were also evaluated for their support of the ECC production. Conditions generally favorable for growth of E. coli O157:H7 were a rich nutrient medium (TSA) vs. heated lettuce juice agar (HLJA) or minimal salts medium (MSM), 22 degrees C compared to 12 degrees C, and an aerobic atmosphere compared to modified atmosphere (1% O(2), 10% CO(2), and 89% N(2)). Conditions favorable for production of ECC on a per cell basis were HLJA, 12 degrees C, and an aerobic atmosphere. This suggests that modified atmosphere packaging of lettuce may not only decrease the growth of E. coli O157:H7 but also its propensity to form biofilm. There was a negative relationship between cell growth and production of ECC on a per cell basis, and environmental conditions that affected the total amount of ECC produced based on initial population reflected a combination of environmental conditions influencing both cell growth and ECC production on a per cell basis. A relative growth index factor (RGIF) was calculated to better understand ECC production as affected by various environmental conditions simultaneously. The production of ECC on a per cell basis by strain 43895-EPS showed a negative linear relationship with pH of TSA at both 12 and 22 degrees C. This strain generally produced a greater amount of ECC on fresh juice agar than on TSA at the same pH, but production of ECC on alfalfa sprout juice agar (FJA, pH 6.45) at 22 degrees C was significantly less than on TSA (pH 6.50). This indicates that nutrient limitation is not based only on nutrient availability. There may be other factors that repress the production of ECC on FJA, and the effects of those factors may be temperature dependent. Further studies will be required to better understand the relationship between nutrient availability and other factors on the production of ECC by E. coli O157:H7 on raw produce.

Biofilms↗

Metabiotic associations of molds and Salmonella Poona on intact and wounded cantaloupe rind.

Salmonella Poona, a serotype rarely implicated in human infections, has recently caused several cantaloupe-associated outbreaks of salmonellosis. Metabiotic associations of molds and foodborne pathogens on produce have been reported. We tested proteolytic activity and measured changes in the pH of cantaloupe rind caused by growth of Alternaria alternata, Cladosporium cladosporioides, Epicoccum nigrum, Geotrichum candidum, and Penicillium expansum. Survival and growth characteristics of Salmonella Poona co-infected with each mold on the surface rind and in wounded rind tissue as affected by temperature were determined. C. cladosporioides, G. candidum, and P. expansum, but not A. alternata and E. nigrum, showed proteolytic activity on agar media containing gelatin and/or casein, with concurrent increases in pH, thus favoring survival and growth of salmonellae. Intact and mechanically wounded tissue of cantaloupe rinds were inoculated with a five-strain mixture of S. Poona and/or test mold. Five inoculation schemes were used: mold only, S. Poona only, mold and S. Poona simultaneously, mold then S. Poona 3 days later, and S. Poona then mold 3 days later. The pH of cantaloupe rinds inoculated with molds and stored at 20 degrees C for 14 days was significantly higher (P < or =0.05) than on day 0. Only the pH of rinds inoculated with C. cladosporioides or G. candidum was significantly higher (P < or =0.05) on day 21 than on day 0, when cantaloupes were stored at 4 degrees C. An initial population of S. Poona increased from 3.3 log(10) cfu/sample (ca. 7 cm(2)) of cantaloupe rind to populations as high as 9.5 log(10) cfu/sample during storage at 20 degrees C for up 14 days, regardless of co-inoculation with molds. Populations of S. Poona decreased or remained constant at 4 degrees C for up to 21 days. Results demonstrate that persistence and growth of S. Poona on intact, wounded, and decaying cantaloupe rind are not markedly affected by the presence of molds.

Colony Count, Microbial↗

Contamination of chicken carcasses in Gauteng, South Africa, by Salmonella, Listeria monocytogenes and Campylobacter.

The presence of the foodborne pathogens, Salmonella spp., Listeria monocytogenes and Campylobacter spp., on 99 fresh and frozen chicken carcasses sourced from various retailers in Gauteng, South Africa, was investigated. Using culture methods, 60.6% of the carcasses were found to be contaminated with one or more pathogens, with 19.2%, 19.2% and 32.3% of the carcasses being found to harbour Salmonella, L. monocytogenes and Campylobacter, respectively. The extent of contamination with one or more pathogens was not significantly different (p>0.1) between fresh or frozen samples or between samples from butcheries, supermarkets or street vendors. Significantly more (p<0.1) fresh carcasses from butcheries than from other outlets were contaminated with Salmonella, while more fresh carcasses from supermarkets were contaminated with Campylobacter. The proportion of carcasses with L. monocytogenes from all sources were similar. Polymerase chain reaction (PCR) results indicate an even higher extent of pathogen contamination, but the PCR techniques need to be further refined before they can be used routinely.

Animals↗

Development of a global stochastic model relating the distribution of individual cell and population physiological states.

Our ability to predict the lag (lambda) prior to growth of foodborne pathogens is limited by our lack of understanding of the physiological changes taking place in the individual cell during the adaptation process. Theoretical models have been developed to describe the stochastic nature of individual cells, and probability distributions have been used to assign hypothetical values of the physiological state to individual cells (p(i)). The aim of this study is to develop a polynomial model which will link distributions of p(i) values to the physiological state of the population (h(0)), and thus to the lambda. Risk analysis software was used to simulate values of p(i) for populations of cells drawn from lognormal distributions with parameters alpha and beta, and growth curves were simulated using a modified continuous-discrete-continuous (CDC) model. Values for h(0) were then obtained for each growth curve by fitting with the heterogeneous population model (HPM). Multiple regression analysis was used to develop a polynomial function which described the subsequent h(0) value as a function of alpha and beta (R(2)=0.9957). Outputs from simulations using the polynomial model agree well with results from related stochastic models, and suggest that distributions can accurately describe the physiological state of cell populations.

Adaptation, Physiological↗

Effect of prior growth temperature, type of enrichment medium, and temperature and time of storage on recovery of Listeria monocytogenes following high pressure processing of milk.

A five-isolate cocktail of Listeria monocytogenes (10(3) cfu/ml in skim or whole raw milk) was subjected to 450 MPa for 900 s or 600 MPa for 90 s. The effects of prior growth temperature, type of milk (skim vs. whole), type of recovery-enrichment media (optimized Penn State University [oPSU] broth, Listeria Enrichment Broth [LEB], Buffered LEB [BLEB], Modified BLEB [MBLEB], and milk), storage temperature and storage time on the recovery of L. monocytogenes were examined. Optimized PSU broth significantly increased the recovery of L. monocytogenes following high pressure processing (HPP), and was 63 times more likely to recover L. monocytogenes following HPP, compared to LEB, BLEB and MBLEB broths (p<0.05; Odds Ratio=63.09, C.I. 23.70-167.96). There was a significant main effect for prior growth temperature (p<0.05). However, this relationship could not be interpreted given the significant interaction effects between temperature and both pressure and milk type. HPP-injured L. monocytogenes could be recovered using both LEB and oPSU broths after storage of milk at 4, 15 and 30 degrees C, with recovery being maximal after 24 to 72 h of storage; however, recovery yield dropped to 0% after prolonged storage of milk at 4 and 30 degrees C. In contrast, storage of milk at 15 degrees C yielded the most rapid rate of recovery and the highest recovery yield (100%), which remained high throughout the 14 days of storage at 15 degrees C. The above factors need to be taken into consideration when designing challenge studies to insure complete inactivation of L. monocytogenes and possibly other foodborne pathogens during high pressure processing of foods.

Animals↗

Model for the combined effects of temperature, pH and sodium chloride concentration on survival of Shigella flexneri strain 5348 under aerobic conditions.

Shigella is recognized as a major foodborne pathogen; however, relatively few studies have been reported on its growth and survival characteristics, particularly under conditions relevant to food. A fractional factorial design was used to measure the effects and interactions of temperature (4-37 degrees C), pH (2-6) and NaCl (0.5-9%) on survival kinetics of Shigella flexneri strain 5348 in BHI broth. Stationary-phase cells were inoculated into sterile media to give initial populations of 6-7 log(10) CFU/ml and bacterial populations were determined periodically by aerobic plate counts. A total of 267 cultures, representing 83 variable combinations of temperature, pH and NaCl concentration, were analyzed. Survivor curves were fitted from plate count data by means of a two-phase linear model to determine lag times and slopes of the curves, from which decimal reduction times (D-values) and times to a 4-log10 inactivation (t 4D) were calculated. Second order response surface models in terms of temperature, initial pH and NaCl concentration were obtained for the inactivation kinetics parameters of S. flexneri using regression analysis. The use of log10 transformation of the inactivation kinetics parameters yielded models with R2 values of >0.8. These models can provide an estimate of Shigella inactivation. The data obtained suggest that Shigella is resistant to acid and salt and that low pH foods stored at low temperatures may serve as vehicles for gastrointestinal illness.

Colony Count, Microbial↗