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1.
2.
Phylogenetic analyses were carried out on a total of 118 Listeria monocytogenes isolates from foods or food processing environments, and 7 isolates from listeriosis patients in Japan to evaluate the genetic variation in the pathogen in this country. Isolates of serotypes 1/2a, 1/2b and 4b were mainly examined to assess the risk of exposure of humans to L. monocytogenes from foods in Japan. The nucleotide sequences of the part of the iap gene that contains the region encoding the threonine-asparagine repeat units were determined in order to construct phylogenetic trees of the isolates investigated. A phylogram showed high genetic diversity among lineage 2 isolates, while the lineage 1 isolates showed clonal characteristics. The results of the genetic analyses suggested the presence of rare putative lineage 3 isolates and epidemic clone I (ECI) isolates in foods in Japan. The results showed that ECI was also isolated from listeriosis patients. The genetic variation in L. monocytogenes in Japan reported here suggests the necessity of monitoring the pathogen in foods and environments in addition to surveillance of listeriosis patients.  相似文献   

3.
One hundred one strains of Listeria monocytogenes isolated from seafood and cheese industry samples and from patients with listeriosis were assessed using a microtiter plate method for adhesion to polystyrene and stainless steel surfaces. The adhesion rate for these strains ranged from 3.10 to 35.29% with an inoculum of 8 x 10(8) cells per well. A strong correlation was found between adhesion to polystyrene and stainless steel microtiter plates, indicating that the intrinsic ability of L. monocytogenes to adhere to inert surfaces is stronger than the influence of the surface's physicochemical properties. The clinical strains were less adherent to inert surfaces than were the industrial strains. By integrating other factors such as location of the industrial strains, contamination type of the clinical strains, serotype, and pulsotype into the analysis, some weak but significant differences were noted. For the industrial isolates, the number of cells attached to both surfaces differed significantly depending on whether they were isolated from food or food-processing environments in the seafood and cheese industry. For clinical isolates, sporadic strains exhibited greater adhesion to polystyrene than did epidemic strains. Strains belonging to the pulsed-field gel electrophoretype clusters A and M (lineages II and I, respectively) were less able to adhere to polystyrene and stainless steel than were strains in the more common clusters.  相似文献   

4.
The aim of this study was to investigate the potential effect of adaptive stationary phase acid tolerance response (ATR) of Listeria monocytogenes Scott A cells on their attachment to stainless steel (SS) under low pH or/and high salt conditions and on the subsequent resistance of sessile cells to strong acid challenge. Nonadapted or acid-adapted stationary-phase L. monocytogenes cells were used to inoculate (ca. 10? CFU/ml) Brain Heart (BH) broth (pH 7.4, 0.5% w/v NaCl) in test tubes containing vertically placed SS coupons (used as abiotic substrates for bacterial attachment). Incubation was carried out at 16 °C for up to 15 days, without any nutrient refreshment. L. monocytogenes cells, prepared as described above, were also exposed to low pH (4.5; adjusted with HCl) or/and high salt (5.5% w/v NaCl) stresses, during attachment. On the 5th, 10th and 15th day of incubation, cells attached to SS coupons were detached (through bead vortexing) and enumerated (by agar plating). Results revealed that ATR significantly (p<0.05) affected bacterial attachment, when the latter took place under moderate acidic conditions (pH 4.5, 0.5 or 5.5% w/v NaCl), with the acid-adapted cells adhering slightly more than the nonadapted ones. Regardless of acidity/salinity conditions during attachment, ATR also enhanced the resistance of sessile cells to subsequent lethal acid challenge (exposure to pH 2 for 6 min; pH adjusted with either hydrochloric or lactic acid). The trend observed with viable count data agreed well with conductance measurements, used to indirectly quantify remaining attached bacteria (following the strong acid challenge) via their metabolic activity. To sum, this study demonstrates that acid adaptation of L. monocytogenes cells during their planktonic growth enhances their subsequent attachment to SS under extended exposure (at 16 °C for up to 15 days) to mild acidic conditions (pH 4.5), while it also improves the resistance of sessile cells to extreme acid treatment (pH 2). Therefore, the ATR of bacterial cells should be carefully considered when applying acidic decontamination strategies to eradicate L. monocytogenes attached to food processing equipment.  相似文献   

5.
The ability of 30 Listeria monocytogenes strains, 15 of meat origin and 15 of clinical origin, to use carnitine as an osmoprotectant and to resist acid stress was determined. All strains examined were able to use carnitine as an osmoprotectant, indicating the importance of this characteristic to the survival of L. monocytogenes in natural environments. Clinical and meat strains, however, differed with respect to this characteristic. Specifically, 73% of meat strains reached a lower maximum cell density in the presence of carnitine with osmotic stress than in its absence with no stress. Only 33% of clinical strains displayed the same feature whereas the remaining clinical strains reached a higher maximum cell density in the presence of carnitine with osmotic stress than in its absence with no stress. The physiological reasons and advantage of this difference are unclear. When exposed to conditions of severe acid stress (pH 2.5) for 2 h, only two L. monocytogenes strains (L66 and L78), both of meat origin, displayed significant reductions (P < 0.05) in number (3.51 and 2.79 log cfu, respectively). Acid-sensitive strains were not found among the clinical isolates examined, highlighting the importance of acid stress resistance in the infection process.  相似文献   

6.
Listeria monocytogenes is an important pathogen responsible for major outbreaks associated with food products. Adhesion to surfaces leads to significant modifications in cell physiology. The aim of this work was to determine the adhesion ability of 10 isolates of L. monocytogenes to eight materials commonly used in kitchens and to evaluate the viability of the adhered cells. The materials assayed were stainless steel 304, marble, granite, glass, polypropylene from a bowl and from a cutting board, and two kinds of silestone. All L. monocytogenes strains attached to all surfaces, although to different extents. L. monocytogenes adhered most tightly to granite and marble, followed by stainless steel 304, glass, silestones, and finally polypropylene surfaces. Surfaces at the threshold between hydrophobicity and hydrophilicity, with high electron acceptor capability and a regular pattern of roughness, were more prone to attachment. Polypropylene surfaces displayed the highest percentage of viable bacteria (nearly 100%), whereas marble and granite had a lower percentage of cultivable cells, 69.5 and 78.7%, respectively. The lowest percentage of culturable bacteria was found on white silestone (18.5%). These results indicate that there are differences in adhered cell viability on different materials. Cell viability assays are important to better understand the cross-contamination process because only adhered bacteria that remain viable are responsible for postprocess contamination.  相似文献   

7.
The survival and growth of Listeria monocytogenes and L. innocua on ready-to-use (RTU) packaged vegetables (lettuce, swedes, dry coleslaw mix, bean-sprouts) were studied. The effects of acid adaptation of Listeria spp. on their survival during subsequent storage were also investigated. Listeria innocua behaviour was similar to that of L. monocytogenes on all vegetables examined. The survival and growth patterns of Listeria varied with the packaged product. Populations on packaged lettuce and swedes significantly increased ( P  < 0.05, by 1–1.5 log cycles) during a 14-day storage period. During the same period, Listeria counts gradually decreased (by 1–1.5 log cycles) on coleslaw mix. Acid adaptation enhanced survival of Listeria spp. during storage in packages of vegetables which had relatively high in-pack CO2 levels (i.e. 25% in packaged coleslaw, bean-sprouts). It is concluded that adapting listerial cells to mildly acidic conditions rendered cultures more resistant to relatively high (25–30%) CO2 atmospheres.  相似文献   

8.
Some subtypes of Listeria monocytogenes can persist in the food-processing industry, but the reasons for such persistence are not known. In the present study, 10 strains of L. monocytogenes representing known persistent randomly amplified polymorphic DNA (RAPD) types from fish processing plants were compared to eight strains of different RAPD type and origin (clinical, food, and animal). All 18 strains of L. monocytogenes had similar growth patterns at different temperatures (5 or 37 degrees C) or different salinities (0.5 or 5% NaCl), and all strains formed a thin layer of adhered cells on a plastic surface when cultured in tryptone soya broth (TSB) with a total of 1% glucose. Many ready-to-eat foods, such as cold-smoked fish, contain NaCl at concentrations of 2 to 5%, and NaCl is present in the processing environment. Adding NaCl to TSB changed the adhesion patterns of all strains, and all adhered better when NaCl was added. Also, the addition of NaCl caused a marked aggregation of 13 of the strains; however, 5 of the 18 strains did not aggregate in the presence of up to 5% NaCl. The aggregates stuck to the plastic surface, and this occurred in all but one of the persistent RAPD types. Four strains represented one particular RAPD type that has been isolated as a persistent RAPD type in several fish processing plants for up to 10 years. Because this RAPD type often can contaminate fish products, it is important to address its potential virulence. The 18 strains differed markedly in their ability to invade Caco-2 cells, and the four strains representing the universal persistent RAPD type were the least invasive (10(2) to 10(3) CFU/ml), whereas other strains invaded Caco-2 cells at levels of 10(4) to 10(5) CFU/ml. Five of the 18 strains belonged to the genetic lineage 1 and were the most invasive. Although the most commonly isolated persistent RAPD type was low invasive, it is important to understand why moderate salinity facilitates aggregation and biofilm formation, for this understanding can be beneficial in developing procedures to reduce processing plant contamination.  相似文献   

9.
Enterococcus faecalis INIA 4 produced the bacteriocin enterocin 4 during growth in raw ewe's milk at 30 degrees C. Enterocin activity reached 2,200 to 3,600 AU/ml after 8 h, with a 1 to 8% (vol/vol) level of inoculum from an 18-h culture. An enterocin activity of 500 AU/ml significantly decreased counts of Listeria monocytogenes Ohio when incubated for 6 h in a model system consisting of filtrates from cultures of E. faecalis INIA 4 in raw ewe's milk, at pH 6.0 and 30 degrees C. However, an enterocin activity of 2,400 AU/ml was needed in the same conditions to significantly decrease counts of L. monocytogenes Scott A. All 22 wild L. monocytogenes strains isolated from ewe's milk and tested were inhibited by a filtrate containing 400 AU/ml of enterocin 4. Incubation in the filtrate for 6 h significantly lowered counts of 16 L. monocytogenes strains, and incubation for 24 h, counts of 21 strains.  相似文献   

10.
Listeria monocytogenes is a Gram-positive foodborne pathogen responsible for a severe disease occurring in immuno-compromised populations. Foodborne illness caused by L. monocytogenes is a serious public health concern because of the high associated mortality. Study of the closely related, but nonpathogenic Listeria innocua has accounted for a better understanding of the behavior of L. monocytogenes in environments beyond the laboratory. Traditionally, the ecological co-habitation, genomic synteny, and physiological similarity of the two species have supported use of L. innocua for predicting the behavior of L. monocytogenes in farm and food processing environments. However, a careful review of the current literature indicates that in a given situation it may not be prudent to use L. innocua as a surrogate for L. monocytogenes without prior confirmation of their similar phenotypes, as an increasing number of studies have arisen demonstrating differences in L. monocytogenes and L. innocua stress response, and furthermore, there are differences among the L. monocytogenes subgroups. Future research should take into consideration that multiple surrogates might be required to accurately model even a single condition depending on the L. monocytogenes subgroup of interest.  相似文献   

11.
Contaminated surfaces of food processing equipment are believed to be a significant source of Listeria monocytogenes to foods. However, very little is known about the survival of Listeria in processing environments. In a mixed bacterial biofilm of L. monocytogenes and Flavobacterium spp., the number of L. monocytogenes cells attaching to stainless steel increased significantly compared to when L. monocytogenes was in a pure culture. The L. monocytogenes cells in the mixed biofilms were also recoverable for significantly longer exposure periods. On colonized coupons held at 15 degrees C and 75% humidity, decimal reduction times were 1.2 and 18.7 days for L. monocytogenes in pure and mixed biofilms, respectively. With increasing exposure time, the proportion of cells that were sublethally injured (defined as an inability to grow on selective agar) increased from 8.1% of the recoverable cell population at day 0 to 91.4% after 40 days' exposure. At 4 and -20 degrees C, decimal reduction times for L. monocytogenes in pure culture were 2.8 and 1.4 days, respectively, and in mixed culture, 10.5 and 14.4 days, respectively. The enhanced colonization and survival of L. monocytogenes on "unclean" surfaces increase the persistence of this pathogen in food processing environments, while the increase in the percentage of sublethally injured cells in the population with time may decrease the ability of enrichment regimes to detect it.  相似文献   

12.
Twenty-nine strains of the foodborne pathogen Listeria monocytogenes were tested for their ability to colonize alfalfa, radish, and broccoli sprouts and their capacity to withstand acid and oxidative stress, two stresses common to the sprouting environment. Wide variation in the ability of different strains to colonize alfalfa sprouts were confirmed, but the variations among radish and broccoli sprouts were not as large. With a few exceptions, strains that were poor colonizers of alfalfa tended to be among the poorer colonizers of radish and broccoli and vice versa. The strains also were variable in their resistance to both acid and oxidative stress. Statistical analysis revealed no correlation between acid stress and sprout colonization, but there was a positive correlation between resistance to oxidative stress and colonization of all three sprout types. Although the response to oxidative stress is important for L. monocytogenes virulence, it also may be important for life outside of a host.  相似文献   

13.
The objective of this study was to evaluate the survival and growth of acid-adapted and nonadapted Listeria monocytogenes inoculated onto fresh beef subsequently treated with acid or nonacid solutions. Beef slices (2.5 by 5 by 1 cm) from top rounds were inoculated with acid-adapted or nonadapted L. monocytogenes (4.6 to 5.0 log CFU/cm2) and either left untreated (control) or dipped for 30 s in water at 55 degrees C, water at 75 degrees C, 2% lactic acid at 55 degrees C, or 2% acetic acid at 55 degrees C. The beef slices were vacuum packaged and stored at 4 or 10 degrees C and were analyzed after 0, 7, 14, 21, and 28 days of storage. Dipping in 75 degrees C water, lactic acid, and acetic acid resulted in immediate pathogen reductions of 1.4 to 2.0, 1.8 to 2.6, and 1.4 to 2.4 log CFU/cm2, respectively. After storage at 10 degrees C for 28 days, populations of L. monocytogenes on meat treated with 55 degrees C water increased by ca. 1.6 to 1.8 log CFU/cm2. The pathogen remained at low population levels (1.6 to 2.8 log CFU/cm2) on acid-treated meat, whereas populations on meat treated with 75 degrees C water increased rapidly, reaching levels of 3.6 to 4.6 log CFU/cm2 by day 14. During storage at 4 degrees C, there was no growth of the pathogen for at least 21 days in samples treated with 55 and 75 degrees C water, and periods of no growth were longer for acid-treated samples. There were no differences between acid-adapted and nonadapted organisms across treatments with respect to survival or growth. In conclusion, the dipping of meat inoculated with L. monocytogenes into acid solutions reduced and then inhibited the growth of the pathogen during storage at 4 and 10 degrees C, while dipping in hot water allowed growth despite initial reductions in pathogen contamination. The results of this study indicate a residual activity of acid-based decontamination treatments compared with water-based treatments for refrigerated (4 degrees C) or temperature-abused (10 degrees C) lean beef tissue in vacuum packages, and these results also indicate that this activity may not be counteracted by prior acid adaptation of L. monocytogenes.  相似文献   

14.
The effects of sodium hypochlorite (NaOCl) and peracetic acid/hydrogen peroxide (PAH) on the inactivation of adherent Listeria monocytogenes LO28 cells were examined. The surfaces tested were stainless steel and polytetrafluoroethylene (PTFE) conditioned or not with an anionic biosurfactant produced by Pseudomonas fluorescens. Dilution-neutralization methods were used to assess the effectiveness of sanitizer solutions on planktonic and adherent cells. Tests were performed on L. monocytogenes cultivated at 37 degrees Celsius (body temperature) or 20 degrees Celsius (ambient temperature). The results demonstrated that i) a total deficiency in nutrients induced by the incubation of cells in 0.15 M NaCl favored the action of NaOCl and PAH on planktonic cells; ii) by reducing the number of cells adhering to stainless steel, pre-conditioning of the surface with the biosurfactant reduced the level of contamination of the surface and thus favored the bactericidal activities of the disinfectants; and iii) the weak binding energies involved in the adsorption of the biosurfactant on PTFE surfaces resulted in there being no reduction by the polymer of the surface contamination. Furthermore, this study confirmed that adherent cells exhibited increased resistance to the actions of the disinfectants when compared to the resistance of planktonic cells.  相似文献   

15.
Comparison of pathogenic bacterial strains of clinical origin with strains of the same species isolated from the environment may be a valuable tool for microbial risk assessment, especially for foodborne pathogens. Thus, a number of Listeria monocytogenes strains responsible for human cases of listeriosis, in relation to the consumption of contaminated seafood, have been compared with "natural" L. monocytogenes strains isolated from similar seafood products. Complete factorial designs were used to assess quantitatively the growth abilities of four clinical and four seafood isolates of L. monocytogenes placed in various environmental conditions. The cells were submitted to acid and osmotic stress as they were in stationary phase (constant condition) or in exponential phase (dynamic condition). The effects and interactions of pH (5-7) and NaCl concentration (0.5-8% v/v) were studied at two growth temperatures (10 and 20 degrees C). Growth parameters (lag and generation times calculated with Gompertz equation) were used to compare the behavior of the strains with respect to the conditions of culture. The results indicated an overall weak effect of acid stress alone, whereas osmotic stress clearly affected bacterial growth and a synergic effect between these two factors was observed. Clinical strains displayed better adaptation than seafood strains in stationary phase, however, this difference was not verified in exponential phase. Low temperature (10 degrees C) usually confirmed the observations at 20 degrees C, and the differences between clinical and food strains were more pronounced. Finally, a classification of the eight strains, based on the collected data, showed three groups: (i) seafood strains, (ii) three clinical strains and (iii) the last clinical strain, alone due to its high resistance to adverse conditions.  相似文献   

16.
The influence of adaptation to habitat structure on the growth of Listeria monocytogenes on processed meats was investigated. Triplicate populations of each of two L. monocytogenes strains were adapted to growth in homogeneous (Tryptic Soy Broth, TSB) or structured (Tryptic Soy Agar) environments by serial transfer in these media incubated at 25 degrees C for 48 h. Adapted (serial transfer for 56 days) and unadapted control (single 48 h growth period) populations were inoculated onto the surface of bologna or farmers' sausage disks, vacuum packaged and stored at 4 degrees C for 4 weeks. Initially and after 4 weeks, L. monocytogenes numbers were determined on Modified Oxford Agar incubated at 25 degrees C for 48 h. Control populations of both strains grown in the structured habitat reached significantly higher levels (1-1.5 log cfu g(-1), P<0.05) on both products than their counterparts grown in homogeneous habitat. Populations of both strains adapted to homogeneous habitats grew to significantly higher levels (0.4-1 log cfu g(-1), P<0.05), while those adapted to growth in structured habitats grew to significantly lower levels (0.3-1 log cfu g(-1), P<0.05), on both products as compared to their unadapted control populations. Populations of both strains adapted to homogeneous habitats grew to significantly higher levels (0.2 to 0.5 log cfu g(-1), P<0.05) on farmers' sausage, and significantly lower levels (0.2 to 0.4 log cfu g(-1), P<0.05) on bologna, than those adapted to structured habitats. Habitat adaptation affected L. monocytogenes growth on processed meats, but the practical significance and reasons for these findings were not established.  相似文献   

17.
《International Dairy Journal》2002,12(2-3):273-283
Minimal processing relies on the use of multiple sub-lethal stresses (or processes) to achieve a similar level of microbial control as that traditionally achieved using a single lethal stress. The benefit to the consumer is products which are less obviously processed than a frozen or canned, acidified or heavily salted food item. However, our increasing understanding of how bacteria can adapt to sub-lethal stresses in a manner which can render them less susceptible to additional insults, should be borne in mind when designing safety or extended shelf-life into a minimally processed product. Listeria monocytogenes is a target organism for many minimally processed food manufacturers because of its ability to tolerate adverse conditions such as low Aw and low temperature. In this communication we use L. monocytogenes as a model system to describe some of the consequences of stress adaptation in terms of improved survival in minimally processed foods and, importantly, the consequences in terms of the virulence of the target organism.  相似文献   

18.
While the majority of human listeriosis cases appear to be linked to consumption of processed ready-to-eat foods (e.g., deli meats), a few listeriosis outbreaks have been linked to consumption of contaminated vegetables. In this study, we assessed four isolates representing the major Listeria monocytogenes lineages for their abilities to attach to and grow on Arabidopsis thaliana, a well-characterized plant model. When plants were dipped for 5min into 3ml of water containing 8.8logCFU of L. monocytogenes and rinsed repeatedly, L. monocytogenes was recovered from the leaves at densities from 1.52 to 2.17logCFU/cm(2). Ten days after exposure, bacterial numbers had increased over initial numbers by 2.60-2.95logCFU/cm(2). Using L. monocytogenes expressing GFP, bacteria were visualized in the intercellular spaces of A. thaliana leaves, suggesting internalization through stomata. These data indicate that L. monocytogenes can rapidly attach to and multiply on plant surfaces and colonize intercellular spaces in A. thaliana leaves where it may be protected from sanitation treatments. When A. thaliana seeds were exposed to L. monocytogenes, between 4.23 and 4.57logCFU/cm(2) were recovered from leaves 7 days post-germination, suggesting that contaminated seeds can produce contaminated plants. Overall, our study demonstrates that prevention of L. monocytogenes contamination of plants throughout growing stages is critical, consistent with recommendations for other produce-transmitted foodborne pathogens.  相似文献   

19.
Wild Listeria isolates representing serovars found in artisanal cheeses commercialized in Asturias (northern Spain) were assessed for their susceptibility to several bacteriocins. Pediocin PA-1 was the most active bacteriocin followed by enterocin AS-48, nisin, and plantaricin C. However, some Listeria monocytogenes and Listeria innocua strains were already highly resistant to PA-1. Among the wild L. monocytogenes populations, the frequency of development of nisin resistance ranged from 10(-6) up to 10(-3), depending on the strain. Highly stable mutants with increased nisin resistance (two- to fourfold) were isolated and tested for potential cross-resistance to lysozyme, EDTA, and various NaCl concentrations and pH values. All mutants were cross-resistant to lysozyme but sensitive to EDTA. In contrast, no clear correlation could be established between nisin resistance and an altered susceptibility to NaCl or pH changes. Nisin-resistant variants were able to survive and even to multiply in milk fermented by a nisin-producing Lactococcus, but the growth of the wild-type strain was inhibited. The different phenotypes evaluated in this study are indicative of the unpredictability of the consequences of the development of nisin resistance in a dairy environment. This resistance should be considered when making a risk assessment of the long-term use of nisin to control L. monocytogenes.  相似文献   

20.
Dairy products, in particular soft cheeses, pose a major concern to the dairy industry and public health authorities as they are the leading source of listeriosis outbreaks, a severe foodborne infection affecting pregnant women, children, elderly and immunocompromised people, with a high (20–30%) mortality rate. Cheeses offer a suitable environment for the survival and growth of Listeria monocytogenes, allowing this pathogen to display tolerance responses that can favour its presence in cheese and persistence in dairy processing plants. Extensive food safety regulations in the EU towards prevention of contamination of dairy products with L. monocytogenes have been implemented. However due to the specific abilities of this pathogen to overcome the processing hurdles, its control remains a challenge. Compliance with the Good Manufacturing Practices, observation of Hazard Analysis Critical Control Points (HACCP) and the surveillance of the pathogen in the cheese processing environment are crucial to provide consumers with a safe product. This review aims to provide an overview on the current knowledge about the potential for the transmission of L. monocytogenes in cheese and its abilities to overcome the challenging processing conditions and implications for the behaviour of the pathogen in the host.  相似文献   

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