Effects of lactic acid and lauricidin on the survival of Listeria monocytogenes,Salmonella enteritidis and Escherichia coli O157:H7 in chicken breast stored at 4 °C

Lauricidin and lactic acid were evaluated for their effects on growth and survival of Listeria monocytogenes (L55), Salmonella enteritidis (S552) and Escherichia coli O157:H7 (E19) inoculated onto raw chicken breast. Fresh, raw chicken breasts were purchased immediately after slaughter and transported on ice to the laboratory within 20 min. Each chicken breast was decontaminated by briefly dipping in 70% ethanol and passed through a flame of a Bunsen burner and then allowed to cool. The decontaminated Chicken breast was dipped in TSB broth, at room temperature (25 °C) for 15 min, containing approximately log 9 CFU/ml of L. monocytogenes, S. enteritidis or E. coli O157:H7. Initial counts of L. monocytogenes, S. enteritidis or E. coli O157:H7 counts in chicken breast immediately after dipping in TSB broth were in the range of log 7–log 8 CFU/g. After inoculation, the chicken breasts were kept at room temperature for 20 min to allow attachment. Each inoculated chicken breast (25 °C) was dipped in 0 (control – sterile water), 0.5%, 1%, 1.5% or 2% of lauricidin (w/v) or lactic acid (v/v) for 10, 20 or 30 min and then individually placed in oxygen-permeable polyethylene bags. Breasts were subjected to microbiological analyses after treatment (day 0) and after storage for 2, 5, 7, 10 and 14 d at 4 °C. Initial counts of L. monocytogenes, S. enteritidis and E. coli O157:H7, in chicken breast treated with lauricidin decreased by 2.90, 1.31 and 2.27 log CFU/g, respectively. Lauricidin was more effective in reducing L. monocytogenes population than S. enteritidis and E. coli O157:H7 population. Dipping chicken breast in lauricidin for 30 min caused a significant reduction of L. monocytogenes, S. enteritidis and E. coli O157:H7 population compared to 10 and 20 min dipping. Initial L. monocytogenes, S. enteritidis and E. coli O157:H7 counts on chicken breast treated with lactic acid decreased by 1.97, 1.71 and 2.59 log CFU/g, respectively. Lactic acid caused a higher reduction in initial S. enteritidis and E. coli O157:H7 counts compared to lauricidin.     

Effect of organic acids,hydrogen peroxide and mild heat on inactivation of Escherichia coli O157:H7 on baby spinach

Minimally processed baby spinach contaminated with Escherichia coli O157:H7 has been associated with multiple outbreaks of foodborne illnesses recently. Chlorinated water is widely used to wash vegetables commercially, but this washing procedure has limited efficacy and can lead to the formation of carcinogenic substances. This study was conducted to determine the effects of organic acids and hydrogen peroxide alone and in binary combinations with or without mild heat (40 and 50 °C) on the inactivation of Escherichia coli O157:H7 on baby spinach. Baby spinach leaves were dip-inoculated with E. coli O157:H7 to a level of 6 log CFU/g and stored at 4 °C for 24 h before treatment. Individual washing solutions (1% and 2% lactic acid [LA], citric acid [CA], malic acid [MA], tartaric acid [TA], acetic acid [AA], hydrogen peroxide [H₂O₂] as well as binary combinations of LA, CA, MA and H₂O₂ at final concentrations of 1% were used to decontaminate spinach leaves at 22, 40 or 50 °C for 2–5 min to test their efficacy in reducing E. coli O157:H7. Chlorinated water (200 ppm free chlorine) decreased the population of E. coli O157:H7 on baby spinach by only 1.2–1.6 log CFU/g, which was not significantly different from DI water washing. Washing with 1% LA at 40 °C for 5 min was the most effective treatment achieving a 2.7 log reduction of E. coli O157:H7 which is significantly higher than chlorine washing. Washing with LA + CA or LA + HP at 40 °C for 5 min was equally effective against E. coli O157:H7, resulting in a 2.7 log reduction of E. coli O157:H7. The application of mild heat significantly enhanced the efficacy of washing solutions on the inactivation of E. coli O157:H7. There was, however, no significant difference between treatments at 40 °C for 5 min and 50 °C for 2 min. The results suggested that the use of organic acids in combination with mild heat can be a potential intervention to control E. coli O157:H7 on spinach.     

Effect of combined ozone and organic acid treatment for control of Escherichia coli O157:H7 and Listeria monocytogenes on enoki mushroom

This study evaluated the effects of ozonated water (1, 3, and 5 ppm) alone with different exposure times (0.5, 1, 3, or 5 min), and combinations of 3 ppm ozone with 1% organic acids (acetic, citric, or lactic acids) during 5-min exposure for control of Escherichia coli O157:H7 and Listeria monocytogenes on enoki mushroom and to observe the regrowth of these pathogenic bacteria on treated enoki mushroom during storage for 10 days at 15 °C. Results showed that ozone treatment gave less than 1.0- and 0.5-log count reductions on E. coli O157:H7 and L. monocytogenes, respectively. Efficacy was improved with combined 3 ppm ozone and 1% citric acid treatment, resulting in 2.26- and 1.32-log count reductions, respectively. During storage at 15 °C (10 days) after combined treatment and packaging, populations of E. coli O157:H7 and L. monocytogenes increased to more than 8.0 log cfu/g, indicating that the combined treatment did not have a residual antimicrobial effect during storage. Although the storage study did not show control of these pathogens, the combined ozone–organic acid treatment was more effective than individual treatments in reducing initial population levels of these pathogens on enoki mushroom.     

Inactivation of Salmonella,E. coli and Listeria monocytogenes in phosphate-buffered saline and apple juice by ultraviolet and heat treatments

Two strains of Escherichia coli (K-12 and O157:H7), Salmonella (enteritidis and typhimurium) and Listeria monocytogenes (AS-1 and M24-1) were individually suspended in phosphate-buffered saline (PBS) and apple juice prior to exposure to UV radiation (220–300 nm) and heating at 55 °C. The calculated decimal reduction times (D value, min) varied with suspending medium and mode of inactivation. The AS-1 and M24-1 strains of L. monocytogenes were found to be most resistant to UV in PBS (0.28–0.29 min) while the AS-1 strain was most resistant in juice (1.26 min). The AS-1 strain of L. monocytogenes and E. coli O157:H7 were most heat resistant when suspended in PBS (4.41 min) and juice (4.43 min), respectively. Results obtained from this study may be used by apple juice processors in selecting appropriate organisms for UV irradiation or heat treatment lethality validations.     

Evaluation of chlorine,benzalkonium chloride and lactic acid as sanitizers for reducing Escherichia coli O157:H7 and Yersinia enterocolitica on fresh vegetables

The effectiveness in the assurance of fresh vegetable microbiological quality of wash solutions containing 200 ppm free chlorine, 0.1 mg/ml benzalkonium chloride, 0.2% and 1% lactic acid was assessed on Escherichia coli O157:H7 and Yersinia enterocolitica contaminated lettuce and tomatoes. Y. enterocolitica reduction on tomatoes (5.08, 4.77 and 4.21 log after 0.2% lactic acid, 200 ppm chlorine and 0.1 mg/ml benzalkonium chloride treatments, respectively) were significantly higher than those for Y. enterocolitica on lettuce and E. coli O157:H7 on both vegetables. Antimicrobial treatment effects on bacterial counts and product quality after subsequent 7 day storage (4 °C and 22 °C) were determined. No pathogens were found in natural microflora of fresh vegetables.     

Survival of bacteria in less than thorough cooked,brine-injected steaks

Bacteria that may include enteric pathogens are introduced into muscle tissues when they are injected with brine. Consequently, steaks of such non-intact, non-comminuted tissues should be cooked to temperatures sufficient to render them microbiologically safe. Studies were conducted to identify the minimum safe temperature for cooking those meats. Suspensions of Escherichia coli and Listeria innocua at numbers about 8 log cfu/ml, in broth or in brines containing 2% or 5% of each of NaCl and sodium tripolyphosphate that were not supplemented or were supplemented with 2% soy protein or 2% emulsified sunflower oil were injected into the centres of 3 cm thick steaks. Groups of four steaks injected with each type of suspension were cooked to temperatures at the centres of 50, 55, 60, 65 or 70 °C. Bacteria were recovered from samples from the centres of steaks, on agars that did or did not allow resuscitation of injured cells of each organism. The results indicated that E. coli but not L. innocua was injured by the brines; that supplementing brines with protein or oil did not protect the organisms from injury or inactivation by heating; and that cooking to a central temperature > 60 ? 65 °C was sufficient to inactivate all bacteria in the meat. The centres of steaks were subsequently injected with 0.1 ml portions of five strain cocktails of E. coli O157:H7 or Listeria monocytogenes in broth containing a dye and the bacteria at numbers >8 log cfu/ml. Steaks were cooked to temperatures of 63, 64 or 65 °C, with holding for 0, 1 or 2 min after cooking before excision of all the dyed tissue from each steak and enumeration of bacteria from the tissue on resuscitating media. The results indicated that cooking to 65 °C without holding would be sufficient to reduce numbers of E. coli O157:H7 or L. monocytogenes in non-intact, non-comminuted steaks by ?7 log units.     

Antimicrobial activity of essential oils against Escherichia coli O157:H7 in organic soil

Soil can be a significant source of preharvest contamination of produce by pathogens. Demand for natural pesticides such as essential oils for organic farming continues to increase. We examined the antimicrobial activity of several essential oils against Escherichia coli O157:H7 in soil. Two essential oils (cinnamaldehyde and eugenol), two bio-pesticides (Ecotrol and Sporan) containing essential oils, and an organic acid (acetic acid) at 0.5%, 1.0%, 1.5% and 2.0%, were mixed with organic sandy soil and inoculated with five different strains of E. coli O157:H7. Soils were incubated at room temperature (22 °C) and samples obtained at 1, 7 and 28 days were enumerated to determine survival. E. coli O157:H7 populations in soil were reduced by up to 5 log cfu/g after 24 h incubation at room temperature with 2% cinnamanaldehyde, Ecotrol, Sporan or vinegar. Reduction in E. coli O157:H7 by eugenol was not significantly different from control. Overall, E. coli O157:H7 strain 4406 was the most sensitive of all the five strains tested and cinnamaldehyde was superior to other treatments in reducing E. coli O157:H7 in soil. In general, increases in essential oil concentrations corresponded to reduced survival of E. coli O157:H7 with all oils used in this study. The results suggest that oils can reduce potential contamination of fresh organic produce inadvertently contaminated by soil.     

Efficiency of high pressure treatment on inactivation of pathogenic microorganisms and enzymes in apple,orange, apricot and sour cherry juices

The purpose of this study was to investigate the effect of high hydrostatic pressure with a mild heat treatment on Staphylococcus aureus 485, Escherichia coli O157:H7 933 and Salmonella Enteritidis FDA in apple, orange, apricot and sour cherry juices. The effectiveness of the treatment on polyphenol oxidase activity in apple juice and pectinesterase activity in orange juice were also determined. An inoculum of microorganisms was completely inactivated at 350 MPa and 40 °C in 5 min. The residual polyphenol oxidase activity in apple juice after treatment at 450 MPa and 50 °C for 60 min was obtained as 9 ± 2.2%. The residual pectinesterase activity in orange juice after treatment at 450 MPa and 50 °C for 30 min was determined as approximately 7 ± 1.6%. It compares with 12 ± 0.2% at a treatment of 40 °C and 450 MPa for 60 min. Pressure resistant isoenzymes were thought to be responsible for the final residual activity. The inactivation is irreversible and the enzyme is not reactivated upon storage. High pressure processing constitutes an effective technology to inactivate the enzymes in fruit juices. Pressures higher than 400 MPa can be combined with mild heat (<50 °C) to accelerate enzyme inactivation.     

Combined effects of heat,acetic acid,and salt for inactivating Escherichia coli O157:H7 in laboratory media

Heat, acid, and salt have been commonly used to ensure the microbial safety of foods and are often used in combinations in many food products. When combined, they can produce different results, such as additive, synergistic, and antagonistic effects. However, there has been little investigation into the effect of these combination treatments. Therefore, in this study, the effect of combined treatment of heat, acid, and salt was investigated in laboratory media. All possible paired combinations among three factors, heat (55 °C), acid (0.25% acetic acid, [v/v]) and salt (3%, [w/v]) were tested and compared with individual treatments for killing E. coli O157:H7 in laboratory media. When salt was combined with heat, there was no significant difference in reduction of E. coli O157:H7 (additive effect). However, when acid was combined with heat, there was a higher reduction of E. coli O157:H7 (synergistic effect). When salt was combined with acid treatment, salt gave protection against acid treatment (antagonistic effect), thus, there was lower reduction of E. coli O157:H7 in the combined treatment than in the single acid treatment. Depending on the combination of preserving factors, results were different.     

Pathogenic microorganism survival in dulce de leche

In order to evaluate the survival of Salmonella typhimurium, Listeria monocytogenes, Escherichia coli O157:H7 and Staphylococcus aureus in dulce de leche, aliquots of this sweet were experimentally contaminated with these pathogenic microorganisms at 10³ (C1) and 10¹ (C2) bacterial cells per gram, and later analysed to evaluate microorganism viability after storage for 0, 1, 2, 3, 5, 10, 20 and 30 days. Salmonella and L. monocytogenes were recovered from all aliquots. E. coli O157:H7 and S. aureus were recovered from all aliquots at C1 concentration, and up to the 5th and 10th day, respectively, at C2 concentration. Relevant public-health danger pathogenic microorganisms were found to survive up to 30 days in dulce de leche.     

Efficacy of sanitizers in reducing Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes populations on fresh-cut carrots

Shredded carrots were inoculated with Escherichia coli O157:H7, Salmonella or Listeria monocytogenes and washed for 1 or 2 min with chlorine (Cl; 200 ppm), peroxyacetic acid (PA; 40 ppm) or acidified sodium chlorite (ASC; 100, 200, 500 ppm) under simulated commercial processing conditions. After washed, the carrots were spin dried, packaged and stored at 5 °C for up to 10 days. Bacterial enumeration was significantly (P ⩽ 0.05) reduced by 1, 1.5 and 2.5 log CFU/g after washing with ASC 100, 250 and 500 ppm, respectively. All sanitizers reduced pathogen load below that of tap water wash and unwashed controls. During storage at 5 °C the bacterial load of all treatments increased gradually, but to different extent in different treatments. ASC inhibited bacterial growth more effectively than the other sanitizers and also maintained the lowest pathogen counts (<1 log CFU/g) during storage. Organic matter in the process water significantly (P ⩽ 0.05) reduced the antibacterial efficacy of Cl, but not that of PA or ASC. Therefore, ASC shows the potential to be used as a commercial sanitizer for washing shredded carrots.     

Combined treatment with silver ions and organic acid enhances growth-inhibition of Escherichia coli O157:H7

Individual and combined treatment with silver ions and selected organic acids were evaluated for growth-inhibition of Escherichia coli O157:H7. The concentrations of silver ion were 0.1, 0.3, 0.5, and 1.0 ppm and, citric, lactic, maleic, succinic, and tartaric acids (0.05% and 0.1%) were used. The inhibitory effect was measured during the growth of E. coli O157:H7 in LB broth by optical density at 600 nm. For individual treatments, silver ion at all concentrations and organic acids at 0.1% had a stronger inhibitory effect than untreated control and 0.05% organic acid treatments. For combined treatments, 1.0 ppm of silver ion with 0.1% of organic acids had the greatest inhibitory effect on E. coli O157:H7 growth. With different concentrations, combination with 1.0 ppm of silver ion and 0.05% of organic acids showed greater inhibitory effect than combination with 0.3 ppm of silver ion and 0.1% of organic acids. Combined treatment with silver ion and organic acid showed the enhanced inhibitory effect of E. coli O157:H7 compared to individual treatments. The present study indicated that growth-inhibiting effect of combined treatment was dependant on the concentration of silver ion rather than that of organic acid.     

Antimicrobial resistance of Salmonella spp. and Escherichia coli isolated from table eggs

The antimicrobial sensitivity of Salmonella spp. and Escherichia coli isolated from the shells and contents of table eggs sampled from sale outlets in Trinidad was determined using the disc diffusion method. The phage types of S. Enteritidis isolates, the phenotypic characteristics of E. coli isolates and the presence of O157 strain were also investigated. Of a total of 74 isolates of Salmonella tested, 17 (22.9%) exhibited resistance to one or more of the seven antimicrobial agents used compared with 104 (88.1%) of 118 E. coli isolates. The difference was statistically significant (P < 0.05; X²). For both microorganisms, resistance was relatively high to streptomycin (54.2%) and tetracycline (35.9%) but low to gentamicin (11.5%) and sulphamethoxazole/trimethoprim (9.4%). Only 1 (1.4%) isolate of Salmonella was multi-resistant while 55 (46.6%) of E. coli isolates were resistant to three or more antimicrobial agents. The frequency of resistance to antimicrobial agents amongst both bacteria was not significantly (P > 0.05; X²) affected by the location of isolation on the egg (shell or content) or source of eggs (farms, shopping malls or other retailers). Eight (19.5%) of 41 S. Enteritidis isolates tested were resistant compared to 4 (26.7%) of 15 isolates of S. Ohio. All S. Enteritidis isolates belonged to phage type 1 (PT1) and all E. coli isolates were non-haemolytic, non-mucoid, sorbitol fermenters and non-O157 strains. It was concluded that the relatively high resistance amongst the bacteria tested could pose therapeutic problems in consumers, particularly in egg-borne salmonellosis or colibacillosis.     

Efficacy of aqueous chlorine dioxide and fumaric acid for inactivating pre-existing microorganisms and Escherichia coli O157:H7, Salmonella typhimurium,and Listeria monocytogenes on broccoli sprouts

The combined effect of aqueous chlorine dioxide (ClO₂) and fumaric acid as a chemical treatment to inactivate pre-existing microorganisms was evaluated using broccoli sprouts. Broccoli sprouts were treated with distilled water, 50 ppm ClO₂, 0.5% fumaric acid, and a combination of 0.5% fumaric acid and 50 ppm ClO₂. Treatment with 50 ppm ClO₂ and 0.5% fumaric acid reduced the initial populations of total aerobic bacteria, yeasts and molds, and coliforms in broccoli sprouts by 2.70, 2.46, and 1.71 log CFU/g, respectively. In addition, the combined treatment of 50 ppm ClO₂ and 0.5% fumaric acid reduced the initial populations of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes inoculated on broccoli sprouts by 2.39, 2.74, and 2.65 log CFU/g, respectively, compared to the control. These results suggest that the combination of aqueous ClO₂ and fumaric acid can be useful as a hurdle for extending the shelf life of broccoli sprouts during storage.     

Inactivation effect of newly developed low concentration electrolyzed water and other sanitizers against microorganisms on spinach

The efficacy of newly developed low concentration electrolyzed water (LcEW) was investigated to inactivate the pathogens on spinach leaves as a convenient and safe alternative sanitizer and it was compared to other sanitizers. Spinach leaves were inoculated with Escherichia coli O157:H7 and Listeria monocytogenes and dip treated with deionized water (DIW), LcEW, strong acid electrolyzed water (SAEW), aqueous ozone (AO), 1% citric acid (CA) and sodium hypochlorite solution (NaOCl) for 3 min at room temperature (23 ± 2 °C). For all pathogens, the similar pattern of microbial reduction on spinach was apparent with LcEW and SAEW washing. In the present study, it was found that LcEW inactivated, at maximum, 1.64–2.80 log cfu/g and DIW resulted in lowest reduction, 0.31–0.95 log cfu/g of background or pathogenic microflora present on spinach leaves compared to the unwashed control. The findings of this study indicate that LcEW and SAEW did not differ significantly (P > 0.05) in reducing background or pathogenic microflora on spinach and LcEW may be a promising sanitizer for washing vegetables without environmental pollution instead of using electrolyzed oxidizing (EO) water or SAEW.     

Prevalence of foodborne pathogens in open markets and supermarkets in Thailand

This study was conducted in Thailand (Bangkok and Pathum Thani provinces), from June 2006 to July 2007, in order to assess the prevalence of Listeria monocytogenes, Escherichia coli O157, Salmonella, Shigella and Vibrio parahaemolyticus in foods. Retail raw meats and seafood, including chicken (n = 109), pork (n = 80), beef (n = 108), shrimp (n = 43) and oysters (n = 48), from open markets and supermarkets were analyzed. Salmonella was found in 22 of 61 (36%) open market samples (48% of chicken, none of pork and beef, and 53% of shrimp) and in 12 of 75 (16%) samples from supermarkets (57%, 12%, 24%, 0% respectively). However, a small number of L. monocytogenes were isolated, where 6 of 217 (3%) were samples from open markets (6% of chicken and 3% of pork) and 17 of 171 (10%) were from supermarkets (3% of beef, 4% of chicken, and 32% of pork). In both markets, L. monocytogenes was not detected from shrimps, neither from oysters. E. coli O157, Shigella and tdh-positive V. parahaemolyticus were not isolated in this collection. Several Salmonella and L. monocytogenes isolates were multidrug-resistant. Both markets would need better assessment, since multidrug-resistant strains have been isolated and they may lead to therapeutic failure.     

Antimicrobial activity of clove (Syzgium aromaticum) oil in inhibiting Listeria monocytogenes on chicken frankfurters

The ability of Listeria monocytogenes to survive and grow at refrigeration temperature in some ready to eat (RTE) poultry products is a public health concern. The inhibitory effect of clove oil (1% and 2%, v/w) applied to the surface of RTE chicken frankfurters was determined on seven strains of L. monocytogenes inoculated at low (10²–10³ cfu/g) or high cell numbers (10⁴–10⁶ cfu/g), and stored at 5 °C for 2 weeks or at 15 °C for 1 week. All strains of L. monocytogenes survived and grew on control frankfurters at 5 °C and 15 °C but growth was inhibited under both storage conditions in the presence of either 1% or 2% clove oil. Depending on the sensory considerations, the addition of clove oil to frankfurters may be an effective strategy to control L. monocytogenes in chicken frankfurters.     

Responses of E. coli O157:H7, L. monocytogenes 1/2 c and Salmonella enteritidis to pH,aw and temperature stress combinations

This study compared the responses of Escherichia coli O157:H7, Listeria monocytogenes 1/2 c and Salmonella enteritidis toward different combinations of physicochemical stresses (pH: 3–8; a_w: 0.93–0.99; temperature: 3–62 °C). Results showed that L. monocytogenes generally had lower inactivation rates and was able to exhibit growth in most number of tested combinations, including those with very high and very low factor settings. Temperature introduced the greatest variation in the measured growth and death parameters but the contributions of both pH and a_w in all temperature ranges were also noted. The results of this study may be applied in the selection of appropriate pathogens in the evaluation of safety of foods that are preserved using individual or combined physicochemical factor control.     

Prevalence of foodborne pathogens in Turkish Van otlu (Herb) cheese

The survey was conducted on 50 unripened Van otlu cheese samples obtained in Van and Hakkari markets at retail level to determine the microbial characteristics with special emphasis on Staphylococcus aureus, Escherichia coli, E. coli O157:H7 and Salmonella spp. The results revealed that S. aureus and E. coli were present in extremely high numbers, with a mean 6.10 and 3.68 log CFU/g, respectively. S. aureus was found in all samples ranging from 2.48 to 7.15 log CFU/g and was present in more than 5.0 × 10⁵ CFU/g in 54% of the samples whereas E. coli was found in 62% of the samples. None of the samples contained E. coli O157:H7; but 3 of the 50 samples had Salmonella spp. The results indicate that Van otlu cheese presents a potential hazard for public health; and the necessary precaution will have to be taken to improve the sanitary practices and cheese manufacturing technique.     

Survival and growth of Listeria monocytogenes in lubricants used in the food industry

The survival and growth of three Listeria monocytogenes strains in 10 lubricants (synthetic and mineral-oil based) used in the food industry, and rapeseed oil, was investigated at room temperature (20 °C) and refrigerated (5 °C). Additionally, the transfer of L. monocytogenes from lubricants to stainless steel surfaces and vice versa was investigated. Though the amount of L. monocytogenes in most lubricants, both pure and soiled, decreased significantly (p < 0.05) during the 14 d test period, lubricants may act as sources of contamination on the basis of the results obtained on the survival of L. monocytogenes. In general, temperature had significant effect (p < 0.05) on listericidal effect of lubricants contrary to soiling (p > 0.05), however the effect of both factors was dependent on lubricant (p < 0.05). The results clearly showed that L. monocytogenes survived in synthetic conveyer belt lubricant diluted in water. In addition, L. monocytogenes was transferred significantly (p < 0.05) from stainless steel surfaces into conveyer-belt lubricants and into mineral-oil based hydraulic oil.