Survival of Salmonella Newport in oysters

Salmonella enterica is the leading cause of laboratory-confirmed foodborne illness in the United States and raw shellfish consumption is a commonly implicated source of gastrointestinal pathogens. A 2005 epidemiological study done in our laboratory by Brands et al., showed that oysters in the United States are contaminated with Salmonella, and in particular, a specific strain of the Newport serovar. This work sought to further investigate the host-microbe interactions between Salmonella Newport and oysters. A procedure was developed to reliably and repeatedly expose oysters to enteric bacteria and quantify the subsequent levels of bacterial survival. The results show that 10 days after an exposure to Salmonella Newport, an average concentration of 3.7 × 10³ CFU/g remains within the oyster meat, and even after 60 days there still can be more than 10² CFU/g remaining. However, the strain of Newport that predominated in the market survey done by Brands et al. does not survive within oysters or the estuarine environment better than any other strains of Salmonella we tested. Using this same methodology, we compared Salmonella Newport's ability to survive within oysters to a non-pathogenic strain of E. coli and found that after 10 days the concentration of Salmonella was 200-times greater than that of E. coli. We also compared those same strains of Salmonella and E. coli in a depuration process to determine if a constant 120 L/h flux of clean seawater could significantly reduce the concentration of bacteria within oysters and found that after 3 days the oysters retained over 10⁴ CFU/g of Salmonella while the oysters exposed to the non-pathogenic strain of E. coli contained 100-times less bacteria. Overall, the results of this study demonstrate that any of the clinically relevant serovars of Salmonella can survive within oysters for significant periods of time after just one exposure event. Based on the drastic differences in survivability between Salmonella and a non-pathogenic relative, the results of this study also suggest that unidentified virulence factors may play a role in Salmonella's interactions with oysters.     

Direct detection of E. Coli O157:H7 in selected food systems by a surface plasmon resonance biosensor

The Spreeta^TM, surface plasmon resonance (SPR)-based biosensor, was used to detect Escherichia coli O157:H7 spiked in milk, apple juice and ground beef extract using specific antibodies. In the Spreeta^TM biosensor light from an LED is reflected off a gold surface, and the angle and intensity corresponding to the SPR minimum is measured and represented as a refractive index (RI) change corresponding to the antigen-antibody coupling at the sensor surface. Milk, apple juice, and ground beef patties spiked with E. coli O157:H7, at varying concentrations, were injected on the sensor surface immobilized with antibodies against the pathogen at a rate of 1 ml/min for a total of 2 min. The change in RI due to the binding of O157:H7 corresponding to each concentration was computed as an average of three replications over a 2 min interaction period. Assays were conducted at near real-time and results obtained after about 30 min of sample injection. Sensitivity of the E. coli O157:H7 assay was 10²-10³ colony forming unit (CFU)/ml. The biosensor assay was also specific to E. coli O157:H7 as other organisms (E. coli K12 and Shigella) did not produce any appreciable change in the sensogram. Further experiments are needed to establish well-defined methods for detecting other food-borne pathogens in more complex and specific food matrices.     

Efficacy of neutral electrolyzed water for sanitization of cutting boards used in the preparation of foods

The effectiveness of neutral electrolyzed water (NEW) to sanitize cutting boards used for food preparation was investigated. Cutting boards made of hardwood and bamboo were inoculated with Escherichia coli K12 and Listeria innocua, dried for 1 h, washed, rinsed and sanitized with NEW, sodium hypochlorite (NaClO) solution, or tap water (control). After each washing protocol, surviving bacterial populations were determined. Results showed that both NEW and NaClO sanitizing solutions produced similar levels of bacterial reductions. In manual washing, the population reductions by NEW and NaClO were 3.4 and 3.6 log₁₀ CFU/100 cm² for E. coli, and 4.1 and 3.9 log₁₀ CFU/100 cm² for L. innocua, respectively. In the automatic washing, the reductions by NEW and NaClO were 4.0 and 4.0 log₁₀ CFU/100 cm² for E. coli, and 4.2 and 3.6 log₁₀ CFU/100 cm² for L. innocua, respectively. No significant differences (P > 0.05) were observed in surviving bacteria counts when comparing board material types.     

Resistances to benzalkonium chloride of bacteria dried with food elements on stainless steel surface

To confirm the importance of washing food sediments from the surface of food-related environments, we examined resistances against benzalkonium chloride of pathogenic bacterial (Escherichia coli O26, Pseudomonas aeruginosa and Staphylococcus aureus) cells dried and adhered on stainless steel dishes with milk, beef gravy or tuna gravy. Suspensions (0.1 ml) of these bacteria (8-9 log cfu/ml) were put on a 5 cm ? stainless steel dish and dried at room temperature (20-24 °C) for 90 min in a bio-clean bench with ventilation. Though these bacteria suspended with distilled water decreased 30-40 fold during the drying period, milk and the gravies protected the bacteria. Without the food elements, the adhered E. coli and Stap. aureus were decreased from 6 to<2 log cfu/dish by 0.5 mg/ml benzalkonium chloride (BKC) for 10 min treatment. Although Ps. aeruginosa showed resistance to BKC, the adhered cells were inactivated by 2.0 mg/ml BKC. However, the bactericidal effect disappeared by the food elements, particularly with milk, even at 1.0 and/or 2.0 mg/ml BKC levels. The protective efficiency of milk on bacteria disappeared if washed with water.     

Inactivation of Escherichia coli by ozone treatment of apple juice at different pH levels

This research investigated the efficacy of gaseous ozone on the inactivation of Escherichia coli ATCC 25922 and NCTC 12900 strains in apple juice of a range of pH levels, using an ozone bubble column. The pH levels investigated were 3.0, 3.5, 4.0, 4.5 and 5.0. Apple juice inoculated with E. coli strains (10⁶ CFU/mL) was treated with ozone gas at a flow rate of 0.12 L/min and ozone concentration of 0.048 mg/min/mL for up to 18 min. Results show that inactivation kinetics of E. coli by ozone were affected by pH of the juice. The ozone treatment duration required for achieving a 5-log reduction was faster (4 min) at the lowest pH than at the highest pH (18 min) studied. The relationship between time required to achieve 5 log reduction (t_5d) and pH for both strains was described mathematically by two exponential equations. Ozone treatment appears to be an effective process for reducing bacteria in apple juice and the required applied treatment for producing a safe apple juice is dependant on its acidity level.     

Bacterial inactivation in fruit juices using a continuous flow Pulsed Light (PL) system

In this work, the susceptibility to pulsed light (PL) treatments of both a Gram-positive (L. innocua 11288) and a Gram-negative (E. coli DH5-??) bacteria inoculated in apple (pH = 3.49, absorption coefficient 13.9 cm^− 1) and orange juices (pH = 3.78, absorption coefficient 52.4 cm^− 1) was investigated in a range of energy dosages from 1.8 to 5.5 J/cm². A laboratory scale continuous flow PL system was set up for the experiments, using a xenon flash-lamp emitting high intensity light in the range of 100-1100 nm. The flashes lasted 360 ??s at a constant frequency of 3 Hz.The results highlighted how the lethal effect of pulsed light depended on the energy dose supplied, the absorption properties of liquid food as well as the bacterial strain examined. The higher the quantity of the energy delivered to the juice stream, the greater the inactivation level. However, the absorbance of the inoculated juice strongly influenced the dose deliver and, therefore, the efficiency of the PL treatment. Among the bacteria tested, E. coli cells showed a greater susceptibility to the PL treatment than L. innocua cells in both apple and orange juices. Following treatment at 4 J/cm², microbial reductions in apple and orange juices were, respectively, 4.00 and 2.90 Log-cycles for E. coli and 2.98 and 0.93 Log-cycles for L. innocua.Sublethally injured cells were also detected for both bacterial strains, thus confirming that membrane damage is an important event in bacterial inactivation by PL.     

The effect of diets supplemented with thyme essential oils and rosemary extract on the deterioration of farmed gilthead seabream (Sparus aurata) during storage on ice

The effect on quality were assessed for gilthead seabream fed five different diets: control (basal diet); BHT (basal diet with 200 mg kg⁻¹ of butylated hydroxytoluene); rosemary (basal diet with 600 mg kg⁻¹ of rosemary extract - Rosmarinus officinalis); carvacrol (basal diet with 500 mg kg⁻¹ of essential oil of Thymbra capitata, carvacrol chemotype); and thymol (basal diet with 500 mg g⁻¹ of essential oil of Thymus zygis, subspecies gracilis, thymol chemotype). After 18 weeks of experimentation, the animals were stored on ice at 4 °C for 0, 7, 14, and 21 days. Physical-chemical, microbiological and sensory analyses were carried out at each sampling point to determine the degree of deterioration in the gilthead seabream. Lower indices of oxidation were observed in animals who were administered feeds supplemented with BHT, carvacrol and (to a lesser degree) rosemary. Lower bacteria counts were observed for the BHT and thymol groups, in addition to a slower deterioration in terms of sensory perception. Accordingly, the addition of natural antioxidants to the diet may have an added effect on fish quality, delaying post mortem deterioration.     

Isolation and evaluation of the antioxidant activity of phenolic constituents of the Garcinia brasiliensis epicarp

A new glycosylated biflavonone, morelloflavone-4′″-O-β-d-glycosyl, and the known compounds 1,3,6,7-tetrahydroxyxanthone, morelloflavone (fukugetin) and morelloflavone-7″-O-β-d-glycosyl (fukugeside) were isolated from the epicarp of Garcinia brasiliensis collected in Brazil. The structures of these compounds were established using ¹H and ¹³C NMR, COSY, gHMQC and gHMBC spectroscopy. The compounds exhibited antioxidant activity. The greatest potency was displayed by morelloflavone (2), with IC₅₀ = 49.5 mM against DPPH and absorbance of 0.583 at 400 μg/mL for the reduction of Fe³⁺. The weakest potency was displayed by 1,3,6,7-tetrahydroxyxanthone (1), with IC₅₀ = 148 mM against DPPH and absorbance of 0.194 at 400 μg/mL for the reduction of Fe³⁺.     

Use of mild irradiation doses to control pathogenic bacteria on meat trimmings for production of patties aiming at provoking minimal changes in quality attributes

The objectives of the present work were to assess the use of moderate doses of gamma irradiation (2 to 5 kGy) and to reduce the risk of pathogen presence without altering the quality attributes of bovine trimmings and of patties made of irradiated trimmings. Microbiological indicators (coliforms, Pseudomonas spp and mesophilic aerobic counts), physicochemical indicators (pH, color and tiobarbituric acid) and sensory changes were evaluated during storage. 5 kGy irradiation doses slightly increased off flavors in patties. Two pathogenic markers (Listeria monocytogenes and Escherichia coli O157:H7) were inoculated at high or low loads to trimming samples which were subsequently irradiated and lethality curves were obtained. Provided that using irradiation doses ≤ 2.5 kGy are used, reductions of 2 log CFU/g of L. monocytogenes and 5 log CFU/g of E. coli O157:H7 are expected. It seems reasonable to suppose that irradiation can be successfully employed to improve the safety of frozen trimmings when initial pathogenic bacteria burdens are not extremely high.     

Real-time polymerase chain reaction for the quantitative detection of tetA and tetB bacterial tetracycline resistance genes in food

A new, rapid, sensitive and specific method was developed to directly detect and quantify tetA and tetB in food. Both tet genes are two of the most frequently present tetracycline resistance genes in Gram-negative bacteria. A set of primers and Taqman probes was designed for each gene. The standard curves were performed using Escherichia coli BM13 (C600 RifR)/RP4 and E. coli NCTC 50365, which carry tetA and tetB, respectively. Meat and fish samples inoculated with these reference strains were used as a matrix to construct the standard curves for the analysis of 20 samples of chicken meat and 10 samples of hake (Merlucius merlucius). The limits of detection in pure culture were 5 cfu/mL (0.7 log cfu/mL) in the case of tetA, 50 cfu/mL (1.7 log cfu/mL) for tetB and 5 × 10² cfu/g (2.7 log cfu/g) for both genes in food samples. The results obtained by real-time quantitative polymerase chain reaction (qPCR) were compared to counts of tetracycline-resistant bacteria obtained by plating extracts of poultry and hake samples in culture media supplemented with 16 mg/L of tetracycline. Counts of tetracycline-resistant bacteria obtained by qPCR showed a positive correlation, especially interesting when compared with microbiological counts of tetracycline-resistant Enterobacteriaceae in poultry meat (r = 0.5509) and with tetracycline-resistant mesophilic aerobic bacteria in hake samples (r = 0.7146). The obtained results demonstrate that this method could be a useful tool for the direct quantification of the amount of bacterial strains that carry tetA and/or tetB genes in food samples.     

Pasteurization of grapefruit juice using a centrifugal ultraviolet light irradiator

Studies are lacking on the nonthermal pasteurization of liquid foods using UV irradiators that centrifugally form very thin films to overcome the problem of limited penetration depth of UV. Grapefruit juice inoculated with Escherichia coli or Saccharomyces cerevisiae was processed at the following conditions: UV dose 4.8–24 mJ/cm²; treatment time 3.2 s, cylinder rotational speed 450–750 rpm, cylinder inclination angle 15–45°, outlet temperature 11 °C, and flow rate 300 ml/min, and was stored for 35 days. Appropriate dilutions of the samples were pour plated with TSA and TSA + 3% NaCl for E. coli and Sabouraud dextrose agar (SDA) and SDA + 5% NaCl for S. cerevisiae. Nonthermal UV processing at 19 mJ/cm², 450 rpm and 15° reduced E. coli in grapefruit juice by 5.1 log₁₀. A dose of 14 mJ/cm² reduced S. cerevisiae by 6.0 log₁₀. Inactivation increased linearly with increasing UV dose. The inactivations at 600 and 750 rpm were similar, and were better than at 450 rpm. The results at 30° and 45° were similar, and were better than at 15°. The occurrence of sublethal injury in either microorganism was not detected. Storing UV processed grapefruit juice at 4 and 10 °C reduced the surviving E. coli to below 1 log₁₀ cfu/ml in 14 days. Processing UV juice reduced the population of S. cerevisiae to less than 1 log₁₀ cfu/ml where it remained for 35 days during refrigerated storage. These results suggest that grapefruit juice may be pasteurized using a nonthermal UV irradiator that centrifugally forms a thin film.     

Population dynamics of Escherichia coli inoculated by irrigation into the phyllosphere of spinach grown under commercial production conditions

Recent outbreaks of food-borne illnesses associated with the consumption of fresh produce have increased attention on irrigation water as a potential source of pathogen contamination. A better understanding of the behaviour of enteric pathogens introduced into agricultural systems during irrigation will aid in risk assessments and support the development of appropriate farm-level water management practices. For this reason, the survival dynamics of two nalidixic acid resistant strains of Escherichia coli after their spray inoculation into the phyllosphere and soil of field spinach were examined over two growing seasons. E. coli strains NAR, an environmental isolate, and DM3n, a non-pathogenic serotype O157:H7, were applied at rates of 10⁴ to 10⁷ cfu/100 ml to the fully developed spinach plants that arose subsequent to the harvesting of their upper leafy portions for commercial purposes (secondary-growth plants). After 72 h, E. coli on spinach were reduced by 3-5 logs. Culturable E. coli were recovered from plants up to 6 days post-inoculation. Survival in soil was greater than in the phyllosphere. Under ambient conditions, the mean 72 h first order decay constant computed by Chick's Law was 0.1 h⁻¹. Although light reduction studies indicated UV irradiation negatively influenced the persistence of E. coli, a simple relationship between UV exposure and phyllosphere E. coli densities could not be established. E. coli introduced to the leafy portions of spinach via spray irrigation displayed rapid declines in their culturability under the open environmental conditions experienced during this study. A 6 day period between the last irrigation and harvest would minimize the risks of E. coli survival in the spinach phyllosphere. E. coli NAR was identified as a possible surrogate for the O157:H7 strain, DM3n.     

Antibacterial effect of plant-derived antimicrobials on major bacterial mastitis pathogens in vitro

The objective of this study was to investigate the antimicrobial effect of plant-derived antimicrobials including trans-cinnamaldehyde (TC), eugenol, carvacrol, and thymol on major bacterial mastitis pathogens in milk. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the aforementioned compounds on Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Staphylococcus aureus, and Escherichia coli were determined. In addition, the bactericidal kinetics of TC on the aforementioned pathogens and the persistence of the antimicrobial activity of TC in milk over a period of 2 wk were investigated. All 4 plant-derived molecules exhibited antimicrobial activity against the 5 mastitis pathogens tested, but TC was most effective in killing the bacteria. The MIC and MBC of TC on Staph. aureus, E. coli, and Strep. uberis were 0.1 and 0.45%, respectively, whereas that on Strep. agalactiae and Strep. dysgalactiae were 0.05 and 0.4%, respectively. The MIC and MBC of the other 3 molecules ranged from 0.4 to 0.8% and 0.8 to 1.5%, respectively. In time-kill assays, TC at the MBC reduced the bacterial pathogens in milk by 4.0 to 5.0 log₁₀ cfu/mL and to undetectable levels within 12 and 24 h, respectively. The antimicrobial effect of TC persisted for the duration of the experiment (14 d) without any loss of activity. Results of this study suggest that TC has the potential to be evaluated as an alternative or adjunct to antibiotics as intramammary infusion to treat bovine mastitis.     

Assessment of factors influencing antimicrobial activity of carvacrol and cymene against Vibrio cholerae in food

Carvacrol and cymene, phenolic compounds naturally present in the essential oil of oregano and thyme, were examined for their antimicrobial activity against Vibrio cholerae (ATCC 14033, VC1, and VC7) inoculated in carrot juice. Carvacrol exhibited a dose dependent inhibitory effect on the bacteria. Although cymene did not have antimicrobial activity against the bacteria, it enhanced the inhibitory ability of carvacrol. At 25 °C, the lowest concentrations of carvacrol and cymene required for zero detectable viable count varied depending on bacterial strains; 5 and 5 ppm, respectively, for VC7; 5 and 7.5 ppm, respectively, for VC1; and 7.5 and 7.5 ppm, respectively, for ATCC 14033. This study also examined several factors influencing the antimicrobial activity of carvacrol and cymene against V. cholerae ATCC 14033, including temperature, bacterial cell number, and food substrate. Carvacrol and cymene inhibited the bacterium in carrot juice at 25 °C more efficiently than at 15 and 4 °C. The doses of both compounds required for zero detectable viable count increased as the number of the bacterial cells in the carrot juice increased. The fat content and the complexity of foods were shown to decrease the antimicrobial activity of the compounds.     

Survival of Escherichia coli O157:H7 in channel catfish pond and holding tank water

Survival of Escherichia coli O157:H7 in channel catfish (Ictalurus punctatus), pond and holding tank water was investigated. Water from three channel catfish ponds was inoculated with ampicillin/nalidixic acid-resistant E. coli O157:H7 transformed with a plasmid encoding for green fluorescent protein at 10⁵, 10⁶, and 10⁷ CFU/ml. Samples were taken from surface, internal organs, and skin scrape of fish and pond water for E. coli O157:H7 enumeration on brain heart infusion (BHI) agar containing ampicillin and nalidixic acid. To determine the survival of E. coli O157:H7 in catfish holding tank water from two farmers markets, the water was inoculated with 10⁷E. coli O157:H7 CFU/ml. E. coli O157:H7 were detected by direct plating for 33 and 69 d in pond and holding tank water, respectively. A rapid decrease of the pathogen was observed in the first 2 weeks to reach 2 log CFU/ml. When E. coli O157:H7 was not recovered by direct plating, the pathogen was isolated by enrichment in TSB for approximately another 30 d from pond and holding tank water. The populations of E. coli O157:H7 found in the internal organs and skin scrape were 5.5 log and 2.5 log CFU/ml, respectively. E. coli O157:H7 from internal organs and water were recovered for at least 12 d. Results suggest that E. coli O157:H7 can survive in channel catfish pond and holding tank water and channel catfish may become a potential carrier of the pathogen.     

Inhibition of verocytotoxigenic Escherichia coli by antimicrobial peptides caseicin A and B and the factors affecting their antimicrobial activities

The antimic robial activities of caseicin A and B antimicrobial peptides (AMPs) were assessed against a selection of verocytotoxigenic Escherichia coli (VTEC) strains (n = 11), other bacterial pathogenic and spoilage bacteria (n = 7), using a model broth system. The ability of the AMPs to retain their antimicrobial activities against a strain of E. coli O157:H7 380-94 under various test conditions (pH, temperature, water activity, sodium chloride concentrations, inoculum size and the presence of competitive microflora) was assessed and the minimum inhibitory concentrations (MIC) and number of surviving E. coli O157:H7 calculated. The mean number of VTEC surviving after exposure to 2 mg/ml caseicin A and B was reduced by 4.96 and 4.19 log₁₀ cfu/ml compared to the respective controls. The susceptibility of E. coli O157:H7 to the caseicin AMPs decreased as temperature, pH, water activity and inoculum size were reduced. The presence of sodium chloride (0.5-2.5%) did not affect the activity of caseicin A (p > 0.05), however it did inhibit the activity of caseicin B. The presence of a competitive microflora cocktail did not significantly (p > 0.05) affect the activities of the AMPs for the majority of the concentrations tested. Using a quantitative PCR assay, the levels of verotoxins (vt1 and vt2) expressed by E. coli O157:H7 following exposure to a sub-inhibitory concentration (0.5 mg/ml) of caseicin A showed that the verotoxin levels did not differ from the levels produced by the control cultures. The antimicrobial activity of caseicin A against E. coli O157:H7 was also tested in a model rumen system, however concentrations of ≥ 2 mg/ml did not significantly (p > 0.05) reduce E. coli O157:H7 numbers in the model system over a 24 h period. The application of caseicin AMPs in food and/or animal production may be valuable in combination with other antimicrobials although further research is required.     

Herd-level relationship between antimicrobial use and presence or absence of antimicrobial resistance in gram-negative bovine mastitis pathogens on Canadian dairy farms

Concurrent data on antimicrobial use (AMU) and resistance are needed to contain antimicrobial resistance (AMR) in bacteria. The present study examined a herd-level association between AMU and AMR in Escherichia coli (n = 394) and Klebsiella species (n = 139) isolated from bovine intramammary infections and mastitis cases on 89 dairy farms in 4 regions of Canada [Alberta, Ontario, Québec, and Maritime Provinces (Prince Edward Island, Nova Scotia, and New Brunswick)]. Antimicrobial use data were collected using inventory of empty antimicrobial containers and antimicrobial drug use rate was calculated to quantify herd-level AMU. Minimum inhibitory concentrations (MIC) were determined using Sensititre National Antimicrobial Resistance Monitoring System (NARMS) gram-negative MIC plate (Trek Diagnostic Systems Inc., Cleveland, OH). Isolates were classified as susceptible, intermediate, or resistant. Intermediate and resistant category isolates were combined to form an AMR category, and multivariable logistic regression models were built to determine herd-level odds of AMR to tetracycline, ampicillin, cefoxitin, chloramphenicol, trimethoprim-sulfamethoxazole combination, sulfisoxazole, streptomycin and kanamycin in E. coli isolates. In the case of Klebsiella species isolates, logistic regression models were built for tetracycline and sulfisoxazole; however, no associations between AMU and AMR in Klebsiella species were observed. Ampicillin-intermediate or -resistant E. coli isolates were associated with herds that used intramammarily administered cloxacillin, penicillin-novobiocin combination, and cephapirin used for dry cow therapy [odds ratios (OR) = 26, 32, and 189, respectively], and intramammary ceftiofur administered for lactating cow therapy and systemically administered penicillin (OR = 162 and 2.7, respectively). Use of systemically administered penicillin on a dairy farm was associated with tetracycline and streptomycin-intermediate or -resistant E. coli isolates (OR = 5.6 and 2.8, respectively). Use of cephapirin and cloxacillin administered intramammarily for dry cow therapy was associated with increasing odds of having at least 1 kanamycin-intermediate or -resistant E. coli isolate at a farm (OR = 8.7 and 9.3, respectively). Use of systemically administered tetracycline and ceftiofur was associated with cefoxitin-intermediate or -resistant E. coli (OR = 0.13 and 0.16, respectively); however, the odds of a dairy herd having at least 1 cefoxitin-intermediate or -resistant E. coli isolate due to systemically administered ceftiofur increased with increasing average herd parity (OR = 3.1). Association between herd-level AMU and AMR in bovine mastitis coliforms was observed for certain antimicrobials. Differences in AMR between different barn types and geographical regions were not observed.     

Evaluation of ability of ferulic acid to control growth and fumonisin production of Fusarium verticillioides and Fusarium proliferatum on maize based media

The aim of this study was to evaluate the efficacy of ferulic acid (1, 10, 20 and 25 mM) at different water activity (a_w) values (0.99, 0.98, 0.96 and 0.93) at 25 °C on growth and fumonisin production by Fusarium verticillioides and Fusarium proliferatum on maize based media. For both Fusarium species, the lag phase significantly decreased (p ≤ 0.001), and the growth rates increased (p ≤ 0.001) at the lowest ferulic acid concentration used (1 mM), regardless of the a_w. However, high doses of ferulic acid (10 to 25 mM) significantly reduced (p ≤ 0.001) the growth rate of both Fusarium species, regardless of the a_w. In general, growth rate inhibition increased as ferulic acid doses increased and as media a_w decreased. Fumonisin production profiles of both Fusarium species showed that low ferulic acid concentrations (1–10 mM) significantly increased (p ≤ 0.001) toxin production, regardless of the a_w. High doses of ferulic acid (20–25 mM) reduced fumonisin production, in comparison with the controls, by both Fusarium species but they were not statistically significant in most cases. The results show that the use of ferulic acid as a post-harvest strategy to reduce mycotoxin accumulation on maize needs to be discussed.     

Synergistic antimicrobial effect of pyrophosphate on Listeria monocytogenes and Escherichia coli O157 in modified atmosphere packaged and refrigerated seabass slices

Effect of pyrophosphate (PP) in combination with modified atmosphere (MAP) (80% CO₂, 10% O₂ and 10% N₂) on the survival of Listeria monocytogenes and Escherichia coli O157 inoculated on seabass slices stored at 4 °C was investigated. PP pretreatment showed the synergistic effect on microbiological inhibition with MAP as evidenced by the lowered TVC and LAB counts, compared with samples stored in air and those kept under MAP. Microbiological changes of seabass slices inoculated with different levels of L. monocytogenes or E.coli O157 (10³ and 10⁵ cfu/g) were monitored during storage. PP pretreatment reduced colony count of E. coli O157 and extended the lag phase of L. monocytogenes. Therefore, MAP in combination with PP pretreatment not only retarded microbiological deterioration of seabass slices but also reduced or inactivated some pathogenic bacteria to some extent.     

The effects of X-ray radiation on Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri inoculated on whole Roma tomatoes

In the last two decades several foodborne disease outbreaks associated with produce were reported. Tomatoes, in particular, have been associated with several multi-state Salmonella outbreaks. Inactivation of inoculated Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri on whole Roma tomato surfaces by X-ray at 0.1, 0.5, 0.75, 1.0, and 1.5 kGy was studied. The main purpose of this study was to achieve a 5 log reduction in consistent with the recommendations of the National Advisory Committee on Microbiological Criteria for Foods. Moreover, the effect of X-ray on inherent microflora (mesophilic counts, psychrotrophic counts and yeast and mold counts) of untreated and treated Roma tomatoes, during storage at ambient temperature (22 °C) for 20 days was also determined. Mixtures of three or two strains of each tested organism was spot inoculated (100 μl) onto the surface of Roma tomatoes (approximately 7–9 log per tomato), separately, and air-dried, followed by treatment with X-ray doses at 22 °C and 55–60% relative humidity. Surviving bacterial populations on tomato surfaces were evaluated using a nonselective medium (tryptic soy agar) with a selective medium overlay for each bacteria; E. coli O157:H7 (CT-SMAC agar), L. monocytogenes (MOA), and S. enterica and S. flexneri (XLD). Treatment with X-ray significantly reduced the population of the tested pathogens on whole Roma tomato surfaces, compared with the control. Approximately 4.2, 2.3, 3.7 and 3.6 log CFU reduction of E. coli O157:H7, L. monocytogenes, S. enterica and S. flexneri per tomato were achieved by treatment with 0.75 kGy X-ray, respectively. More than a 5 log CFU reduction per tomato was achieved at 1.0 or 1.5 kGy X-ray for all tested pathogens. Furthermore, treatment with X-ray significantly reduced the inherent microflora on Roma tomatoes. Inherent levels were significantly (p < 0.05) lower than the control sample throughout storage for 20 days.