Variation of antibiotic resistance in Salmonella Enteritidis,Escherichia coli O157:H7, and Listeria monocytogenes after exposure to acid,salt, and cold stress

Bacteria with antibiotic-resistant could seriously threaten to human health, increasing the treatment cost for infections and negatively affecting treatment outcomes. Stress adaptation is one possible mechanism for the acquisition or enhancement of antibiotic resistance in bacteria as a result of cross-protection. In this study, the effects of acid, salt, and cold stress on the antibiotic resistance of Salmonella Enteritidis, Listeria monocytogenes, and Escherichia coli O157:H7 were investigated using the disc diffusion method. For S. Enteritidis, acidic growth conditions increased resistance to ciprofloxacin and erythromycin (p < .05), and addition of 4% NaCl to growth media decreased resistance to chloramphenicol (p < .05). Irrespective of pH and the NaCl concentration of the growth medium, refrigerated E. coli O157:H7 showed increased resistance to amoxycillin, ciprofloxacin, gentamicin, streptomycin, and erythromycin (p < .05). Acid-adapted L. monocytogenes showed decreased the resistance to amoxycillin, ampicillin, chloramphenicol, ciprofloxacin, erythromycin, gentamicin, streptomycin, and tetracycline (p < .05). In conclusion, prolonged exposure of foodborne pathogens to acid, salt, and cold stress alters their antibiotic resistance. However, the effect of acid, salt, and cold stress on bacterial antibiotic resistance depend on both the bacterial species and the specific antibiotic. Therefore, multiple factors need to be considered for a foodborne antimicrobial resistant risk assessment.     

Efficacies of Garlic and L. sakei in Wine‐Based Marinades for Controlling Listeria monocytogenes and Salmonella spp. in Chouriço de Vinho,a Dry Sausage Made from Wine‐Marinated Pork

Chouriço de vinho is made from roughly minced (10 to 30 mm) pork and fat, seasoned with a marinade made from wine, salt, garlic, and other facultative seasonings used according to the recipe of each producer. The batter is maintained at 4 to 7 ºC for 24 to 48 h. It is then stuffed into natural thin pork gut, cold smoked and matured at a low temperature for 1 to 4 wk. The effect of garlic used in wine‐based marinade and a starter culture of indigenous Lactobacillus sakei on the behavior of Listeria monocytogenes and Salmonella spp. in the processing of chouriço was investigated. The garlic (as powder and fresh juice) was found to contribute (P < 0.05) to the control of both pathogens in broth. Garlic dose, as tested within the usual limits used for seasoning, did not impact the reduction of pathogens. Garlic‐wine‐based marinade and a starter culture of indigenous L. sakei contribute to controlling L. monocytogenes and Salmonella spp. in the processing of chouriço. Their presence was responsible for the loss of viability of L. monocytogenes and Salmonella spp. following 5 d of drying, even sooner than situations where no garlic was used. The results of the present work show that the use of a wine‐based marinade with garlic has an important role in ensuring the safety of the product.     

Modeling dependence of growth inhibition of Salmonella Typhimurium and Listeria monocytogenes by oregano or thyme essential oils on the chemical composition of minced pork

The aim of this study was to compare the antimicrobial effect of selected essential oils (EOs) at concentration of 1% in minced pork from five different meat cuts (loin, ham, shoulder, neck, and belly) and to assess the influence of the chemical parameters of meat on EO efficiency, determined by the decrease of pathogens in comparison to a control in samples after storage at 3°C/7 d in a vacuum. The inhibition was significant (p < .05) only in minced pork from meat cuts with a fat content lower than 5% (leg, loin and shoulder). Salmonella enterica serovar Typhimurium was more sensitive than Listeria monocytogenes and oregano was more effective than thyme (p < .001). The inhibition of S. Typhimurium depended mainly on the fat content (logarithmic dependence) and to a lesser degree on the pH, whereas pH was not found to be a significant predictor in the case of L. monocytogenes. The results of this study show that only lean minced meat (with a fat content max. 5%) is a suitable matrix for pathogen control by EOs.     

Control of Growth of L. monocytogenes in Fresh Salmon using Microgard and Nisin

Pathogens like Listeria monocytogenes in fresh, or even in cold smoked salmon, have become a major public health concern for the salmon processing industry and government agencies. The effect of bacteriocin solutions (Microgard^™ and Nisin) on reducing total microbial counts, inhibiting Listeria monocytogenes, and prolonging the shelf-life was evaluated. Listeria monocytogenes was inoculated onto chilled and on frozen and thawed salmon samples. The combination of Nisin and Microgard reduced the total aerobic bacteria populations of fresh chilled salmon by 2 log (P<0.05) and increased its shelf-life, at 6 °C, by 3-4 d, as compared with the control. The above bacteriocin combination also reduced the growth of inoculated Listeria monocytogenes in frozen-thawed salmon and increased its shelf-life from 5 to 10 d at 6 °C. The bacteriocin treatment did not affect surface pH values or color of the fish.     

Antimicrobial activity of whey protein based edible films incorporated with oregano,rosemary and garlic essential oils

The use of edible films to release antimicrobial constituents in food packaging is a form of active packaging. Antimicrobial properties of spice extracts are well known, however their application to edible films is limited. In this study, antimicrobial properties of whey protein isolate (WPI) films containing 1.0–4.0% (wt/vol) ratios of oregano, rosemary and garlic essential oils were tested against Escherichia coli O157:H7 (ATCC 35218), Staphylococcus aureus (ATCC 43300), Salmonella enteritidis (ATCC 13076), Listeria monocytogenes (NCTC 2167) and Lactobacillus plantarum (DSM 20174). Ten millilitres of molten hard agar was inoculated by 200 μl of bacterial cultures (colony count of 1 × 10⁸ CFU/ml) grown overnight in appropriate medium. Circular discs of WPI films containing spice extracts, prepared by casting method, were placed on a bacterial lawn. Zones of inhibition were measured after an incubation period. The film containing oregano essential oil was the most effective against these bacteria at 2% level than those containing garlic and rosemary extracts (P < 0.05). The use of rosemary essential oil incorporated into WPI films did not exhibit any antimicrobial activity whereas inhibitory effect of WPI film containing garlic essential oil was observed only at 3% and 4% level (P < 0.05). The results of this study suggested that the antimicrobial activity of some spice extracts were expressed in a WPI based edible film.     

Thiol oxidation and protein cross-link formation during chill storage of pork patties added essential oil of oregano,rosemary, or garlic

The effect of two levels (0.05% and 0.4%) of essential oil of rosemary, oregano, or garlic on protein oxidation in pork patties was studied during storage under modified atmosphere (MAP: 70% O₂: 20% CO₂: 10% N₂) or under aerobic conditions (AE) at 4 °C. The oxidative stability of the meat proteins was evaluated as loss of thiols for up to 9 days of storage, and as formation of myosin cross-links analyzed by SDS-PAGE after 12 days of storage. Protein thiols were lost during storage to yield myosin disulfide cross-links. Essential oils of rosemary and oregano were found to retard the loss of thiols otherwise resulting in myosin cross-links. Garlic essential oil, on the contrary, was found to promote protein oxidation, as seen by an extreme loss in thiol groups, and elevated myosin cross-link formation compared to control.     

Synergistic effect of peppermint and garlic essential oil on fate of Kluyveromyces marxianus in Zucchini Bouranee

The purpose of the study was to evaluate the inhibitory effect of garlic (GEO) and peppermint (PEO) essential oils on Kluyveromyces marxianus (K. marxianus) in zucchini bouranee and also to assess the product's functional physicochemical properties during storage. The results showed that the combined use of the GEO and PEO caused a faster decrease in the yeast count. The growth rate of K. marxianus reduced significantly with increasing the EOs concentration (p < .05). No significant difference was found between amounts of the dry matter of the treatments until the 14th day. The syneresis percentages were significantly different between the treatments during storage. Meanwhile, the GEO treatments had a higher syneresis rate compared with the PEO treatments (p < .05). The organoleptic assays showed that the GEO 0.05% treatment obtained the highest score. As a result, the antimicrobial properties of the EOs, especially GEO, could increase the zucchini bouranee shelf life.     

Incorporation of Essential Oils and Nanoparticles in Pullulan Films to Control Foodborne Pathogens on Meat and Poultry Products

The incorporation of essential oils and nanotechnology into edible films has the potential to improve the microbiological safety of foods. The aim of this study was to evaluate the effectiveness of pullulan films containing essential oils and nanoparticles against 4 foodborne pathogens. Initial experiments using plate overlay assays demonstrated that 2% oregano essential oil was active against Staphylococcus aureus and Salmonella Typhimurium, whereas Listeria monocytogenes and Escherichia coli O157:H7 were not inhibited. Two percent rosemary essential oil was active against S. aureus, L. monocytogenes, E. coli O157:H7, and S. Typhimurium, when compared with 1%. Zinc oxide nanoparticles at 110 nm were active against S. aureus, L. monocytogenes, E. coli O157:H7, and S. Typhimurium, when compared with 100 or 130 nm. Conversely, 100 nm silver (Ag) nanoparticles were more active against S. aureus than L. monocytogenes. Using the results from these experiments, the compounds exhibiting the greatest activity were incorporated into pullulan films and found to inhibit all or some of the 4 pathogens in plate overlay assays. In challenge studies, pullulan films containing the compounds effectively inhibited the pathogens associated with vacuum packaged meat and poultry products stored at 4 °C for up to 3 wk, as compared to control films. Additionally, the structure and cross‐section of the films were evaluated using electron microscopy. The results from this study demonstrate that edible films made from pullulan and incorporated with essential oils or nanoparticles may improve the safety of refrigerated, fresh or further processed meat and poultry products.     

Antimicrobial activity of some plant extracts and essential oils against foodborne pathogens in vitro and on the fate of inoculated pathogens in chocolate

The efficacy of commercially available plant extracts and essential oils used extensively as flavour ingredients in confectionery products were used as antimicrobials in laboratory media against the following microorganisms: Escherichia coli O157:H7, Salmonella Enteritidis, Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus. Using the disc diffusion method, inhibition zones in diameter >20 mm were observed by adding 10 μl of each antimicrobial substance on the following microorganisms: lemon flavour applied on E. coli O157:H7, lemongrass essences against S. aureus, plum using a B. cereus strain and strawberry flavour using a L. monocytogenes strain. E. coli O157:H7 strains were the most susceptible microorganisms inhibited by 18 extracts, followed by S. Typhimurium and S. aureus which were inhibited by 17 extracts. Lemon flavour, lemongrass essences, pineapple and strawberry flavour inhibited the foodborne pathogens at the lowest concentration (5 ml/100 ml). Plant extracts and essential oils with potent antimicrobial activities were tested in chocolate held at different temperatures (7 and 20 °C) in dry or humidified environment, which resulted in different a_w values of the product (i.e. 0.340, 0.450, and 0.822), in order to determine their efficacy on the fate of the inoculated pathogens. The most inhibitory action was observed by lemon flavour applied on chocolate inoculated with E. coli cocktail culture after storage at 20 °C for 9 days. Plant extracts tested on chocolate show an enhanced inhibitory effect during storage at 20 °C indicating that their application may provide protection in case of storage at the above temperature or even higher.     

Biodegradable gelatin–chitosan films incorporated with essential oils as antimicrobial agents for fish preservation

Essential oils of clove (Syzygium aromaticum L.), fennel (Foeniculum vulgare Miller), cypress (Cupressus sempervirens L.), lavender (Lavandula angustifolia), thyme (Thymus vulgaris L.), herb-of-the-cross (Verbena officinalis L.), pine (Pinus sylvestris) and rosemary (Rosmarinus officinalis) were tested for their antimicrobial activity on 18 genera of bacteria, which included some important food pathogen and spoilage bacteria. Clove essential oil showed the highest inhibitory effect, followed by rosemary and lavender. In an attempt to evaluate the usefulness of these essential oils as food preservatives, they were also tested on an extract made of fish, where clove and thyme essential oils were the most effective. Then, gelatin–chitosan-based edible films incorporated with clove essential oil were elaborated and their antimicrobial activity tested against six selected microorganisms: Pseudomonas fluorescens, Shewanella putrefaciens, Photobacterium phosphoreum, Listeria innocua, Escherichia coli and Lactobacillus acidophilus. The clove-containing films inhibited all these microorganisms irrespectively of the film matrix or type of microorganism. In a further experiment, when the complex gelatin–chitosan film incorporating clove essential oil was applied to fish during chilled storage, the growth of microorganisms was drastically reduced in gram-negative bacteria, especially enterobacteria, while lactic acid bacteria remained practically constant for much of the storage period. The effect on the microorganisms during this period was in accordance with biochemical indexes of quality, indicating the viability of these films for fish preservation.     

Optimising chitosan–pectin hydrogel beads containing combined garlic and holy basil essential oils and their application as antimicrobial inhibitor

Chitosan–pectin hydrogel beads that trap and release the maximal amount of combined garlic and holy basil essential oils to inhibit food microorganisms were developed based on the central composite design, with chitosan (0.2–0.7% w/v), pectin (3.5–5.5% w/v) and calcium chloride (CaCl₂) (5.0–20.0% w/v) contents. The optimal bead consisted of 0.3–0.6% w/v chitosan, 3.9–5.1% w/v pectin and 8.0–17.0% w/v CaCl₂, which had a high encapsulation efficiency (62.16–79.06%) and high cumulative release efficiency (31.55–37.81%) after storage at 5 °C for 15 days. Optimal hydrogel beads were packed into a cellulose bag to evaluate antimicrobial activity by the disc volatilisation method. The beads inhibited Bacillus cereus, Clostridium perfringens, Escherichia coli, Pseudomonas fluorescens, Listeria monocytogenes and Staphylococcus aureus but did not affect Lactobacillus plantarum and Salmonella Typhimurium. The oil-containing beads could potentially be applied in food packaging to inhibit the mentioned microorganisms.     

Antimicrobial activities of spice extracts against pathogenic and spoilage bacteria in modified atmosphere packaged fresh pork and vacuum packaged ham slices stored at 4 °C

The antimicrobial activity of 14 spice extracts against four common meat spoilage and pathogenic bacteria (Listeria monocytogenes, Escherichia coli, Pseudomonas fluorescens and Lactobacillus sake) was screened in cultured media (experiment 1). The results showed that individual extracts of clove, rosemary, cassia bark and liquorice contained strong antimicrobial activity, but the mixture of rosemary and liquorice extracts was the best inhibitor against all four types of microbes. Subsequently, mixed rosemary/liquorice extracts were spray-applied to inoculated fresh pork in modified atmosphere packaging (experiment 2) and to inoculated ham slices in vacuum packaging (experiment 3). The meat samples were stored at 4 °C over a 28-day period and microbial growth was monitored regularly. The L. monocytogenes population on fresh pork by day 28 decreased 2.9, 3.1 and 3.6 logs, the MAB decreased 2.7, 2.9 and 3.1 logs, the Pseudomonas spp. count decreased 1.6, 2.1 and 2.6 logs and the total coliform count decreased 0.6, 0.8 and 1.2 logs, corresponding to 2.5, 5.0 and 10.0 mg/ml of spray, respectively, when compared to control (P < 0.05). The number of L. monocytogenes on ham slices decreased 2.5, 2.6 and 3.0 logs, the MAB plate counts decreased 2.9, 3.0 and 3.2 logs and the LAB counts decreased 2.4, 2.6 and 2.8 logs (P < 0.05), respectively, after 28-days, by the same levels of mixed rosemary/liquorice extract treatments. The results demonstrated strong potential of mixed rosemary and liquorice as a natural preservative in fresh pork and ham products.     

Evaluation of pre‐heating and extraction solvents in antioxidant and antimicrobial activities of garlic,and their application in fresh pork patties

The objectives of this study were to screen the optimum conditions for antioxidant and antimicrobial activities of garlic as affected by pre‐heating and different extraction solvents, and to evaluate the antioxidant and antimicrobial effects of these extracts in ground meat during refrigerated storage. Methanol extracted garlic had a greater total phenolic content, 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH)‐radical scavenging activity and reducing power than water extracted one (P < 0.05), whereas the latter had a greater yield and iron chelating ability than the former (P < 0.05). Moreover, water extract from fresh garlic (WEFG) and methanol extract from heated garlic (MEHG) produced an inhibition zone against Escherichia coli O157:H7 and Listeria monocytogenes. The addition of garlic extracts (WEFG, MEHG and their combinations WEFMEHG)) to pork patties decreased the pH, hunter a values (redness), thiobarbituric acid substances values and the number of total plate count and Enterobacteriaceae (P < 0.05), while the hunter b values (yellowness) increased (P < 0.05). Results of this study indicated that the use of the garlic extracts was able to control lipid oxidation and microbial growth in pork patties.     

芳香精油在食品保藏中的应用性研究进展

芳香精油是重要的植物次生代谢物,药食应用历史悠久,因其多方面的生物活性,已引起人们对其在食品保藏领域替代化学防腐剂的极大兴趣。文中概述了芳香精油及其成分在采后果蔬、预切果蔬片、肉制品、乳制品和水产品等食品保鲜和贮藏领域中应用性和安全性研究的最新研究进展。许多研究表明,牛至、丁香、罗勒、胡荽、肉桂、百里香、薄荷、迷迭香、芥茉、芫荽、红根草等芳香精油以及丁子香酚、香芹酚、肉桂酸、己醛、草蒿脑、麝香草酚、香芹酮、肉桂醛、柠檬醛、香叶醇等单体成分具有优良的抗食源性腐败菌和致病菌活性。通过与食品生产前处理工艺、气调包装、配方优化等手段的结合,一些芳香精油及其成分显示出天然食品保藏剂良好的商用价值。     

Evaluation of the effect of defrosting practices of ground beef on the heat tolerance of Listeria monocytogenes and Salmonella Enteritidis

The effect of common defrosting practices of ground beef, including (i) defrosting in the refrigerator (5 °C for 15 h), (ii) defrosting at room temperature (25 °C for 12 h) and (iii) defrosting in the microwave, on the heat tolerance of artificially inoculated Listeria monocytogenes and Salmonella Enteritidis, was studied. The thermal inactivation of S. Enteritidis was not, overall, affected by defrosting practices. In contrast, defrosting at room temperature resulted, overall, in an increased heat tolerance of L. monocytogenes compared to the rest tested defrosting practices. Inactivation kinetics of the two pathogens for the different defrosting practices were determined by fitting the data to the Weibull model. The δ parameter of the Weibull model (heat challenge time (min) required for the first 1-log reduction) for S. Enteritidis and for defrosting at 25 °C, microwave defrosting, defrosting at 5 °C and for the control (fresh ground beef inoculated with the pathogens just before the heat challenge trials) was 1.13, 1.62, 1.60 and 0.96, respectively, while the corresponding values for L. monocytogenes were 20.13, 10.82, 9.95 and 9.47, respectively. The findings of this study should be useful in risk assessments and in developing food handling guidelines for the consumers.     

Antimicrobial activity of recombinant human lactoferrin from <Emphasis Type="Italic">Aspergillus awamori</Emphasis>, human milk lactoferrin and their hydrolysates

The antibacterial activity of human lactoferrin from milk (hLF), recombinant human lactoferrin from Aspergillus awamori (rhLF) and their hydrolysates obtained with pepsin was investigated against Escherichia coli O157:H7, Salmonella Enteritidis and Listeria monocytogenes. The minimum inhibitory concentrations (MIC) and the minimum bactericidal concentrations (MBC) were determined for all the bacteria and the proteins assayed. Taking into account the MICs found for both lactoferrins studied, we can say that they behave very similarly, except for L. monocytogenes for which rhLF was more active. We studied the effect that heat treatments exerted on the antibacterial activity of the two types of lactoferrin and the only heat treatment that had a negative effect on that activity was 85 °C for 10 min. The activity of hLF and rhLF in UHT milk and whey against E. coli O157:H7 and L. monocytogenes, was also assayed. Our results showed a reduction in the number of viable cells for both microorganisms when were incubated with rhLF or hLF, but this decrease was lower than in broth media.     

Effects of sodium hypochlorite and acidified sodium chlorite on the morphological,microbiological, and sensory qualities of selected vegetables

The effects of washing with sodium hypochlorite (NaClO) and acidified sodium chlorite (ASC) on the reduction of various pathogens contaminated produce, as well as the change in sensory qualities during storage and morphological characteristics of the surface of the washed produce, including cherry tomato, cucumber, and carrot were evaluated. Produce was inoculated with Bacillus cereus, B. cereus spore, Campylobacter jejuni, Salmonella Typhimurium, and Listeria monocytogenes and then washed with 100, 150, or 200 ppm NaClO and stored for 11 days at 4°C. The produce contaminated with S. Typhimurium or L. monocytogenes was also washed with 100 ppm NaClO containing 5% acetic acid and 500 ppm ASC. The greatest washing efficacy was observed in B. cereus, followed by S. Typhimurium, L. monocytogenes, and C. jejuni. NaClO with 5% acetic acid was significantly (p<0.05) more effective than 500 ppm ASC at S. Typhimurium, but not at L. monocytogenes, regardless of the types of produce, while ASC was the most effective sanitizer for L. monocytogenes in cucumber. Scanning electron microscopy showed that washing with NaClO removed the pathogens, but also part of the wax layer from the surface, which might affect sensory qualities of the produce during storage.     

Effect of Eryngium caeruleum essential oil on microbial and sensory quality of minced fish and fate of Listeria monocytogenes during the storage at 4°C

The present study investigated in vitro antimicrobial activity of Eryngium caeruleum essential oil (EEO) against five foodborne pathogenic bacteria based on microdilution and disk diffusion methods. Moreover, its effects on specific spoilage microorganisms, inoculated Listeria monocytogenes, and its sensory changes in minced fish were evaluated during 12 days of storage at refrigeration temperature. The results showed that Staphylococcus aureus and Escherichia coli were the most sensitive and the most resistant bacteria with a minimum inhibitory concentration of 0.125 and 1 mg/ml, as well as inhibition zones of 15.66 and 11.66 mm, respectively. Regarding the antimicrobial effect of EEO on the microbial profile and inoculated L. monocytogenes, treating with 0.4% EEO caused a significant decrease in the studied microorganisms when compared to the control group (p < 0.05). In addition, considering the sensory evaluation, the best scores were observed for the samples treated with 0.2% and 0.4% EEO. However, none of the groups obtained acceptable scores until the final day of storage except for the color attribute. In general, sensory evaluation and its correlation with microbial counting indicated that the treatment with 0.4% EEO was able to preserve the microbial quality of the minced fish at refrigeration temperature without any undesirable sensory effects.     

Effect of oregano and thyme essential oils on the microbiological and chemical quality of refrigerated (4 °C) ready‐to‐eat squid rings

The efficacy of oregano and thyme essential oils (OEO and TEO, respectively) in the quality retention of a refrigerated (4 °C) squid (Loligo vulgaris) ring ready‐to‐eat (RTE) product was studied. Essential oils were added at different concentrations to the coating medium during processing. An inhibitory (P < 0.05) effect of OEO on the microbial activity (aerobes, anaerobes, Enterobacteriaceae, psychrotrophs) of the squid rings was observed, with a more pronounced effect as OEO concentration increased. The addition of OEO also led to an inhibitory (P < 0.05) effect on lipid oxidation, as determined by peroxide, thiobarbituric acid‐reactive substance and interaction compound formation; however, no effect (P > 0.05) of the OEO concentrations on lipid oxidation development was detected. The addition of TEO did not lead to an inhibitory effect (P > 0.05) on the microbial activity of the refrigerated RTE squid, although a slight inhibitory (P < 0.05) effect on lipid oxidation was observed in the batches including the higher TEO concentrations.     

Antimicrobial activity of 405 nm light-emitting diode (LED) in the presence of riboflavin against Listeria monocytogenes on the surface of smoked salmon

This study investigated the antimicrobial activity of 405 nm light-emitting diode (LED) with and without riboflavin against Listeria monocytogenes in phosphate buffered saline (PBS) and on smoked salmon at different storage temperatures and evaluated its impact on food quality. The results show that riboflavin-mediated LED illumination in PBS 25 °C significantly inactivated L. monocytogenes cells by 6.2 log CFU/mL at 19.2 J/cm², while illumination alone reduced 1.9 log CFU/mL of L. monocytogenes populations at 57.6 J/cm². L. monocytogenes populations on illuminated smoked salmon decreased by 1.0–2.2 log CFU/cm² at 1.27–2.76 kJ/cm² at 4, 12, and 25 °C, regardless of the presence of riboflavin. Although illumination with and without riboflavin caused the lipid peroxidation and color change in smoked salmon, this study demonstrates the potential of a 405 nm LED to preserve the smoked salmon products, reducing the risk of listeriosis.