Aerobic bacterial, coliform, Escherichia coli and Staphylococcus aureus counts of raw and processed milk from selected smallholder dairy farms of Zimbabwe

A cross sectional study was conducted to enumerate total viable bacteria (TBC), coliforms, Escherichia coli and Staphylococcus aureus in raw (n = 120) and processed (n = 20) milk from individual farms from three smallholder dairy schemes of Zimbabwe between October, 2009 and February, 2010. Data on management factors were collected using a structured questionnaire. A standard pour plate technique was used to enumerate total viable bacteria, while for coliforms, E. coli and S. aureus, counts were assessed by the spread plate technique. The association of total viable bacterial counts and management factors was assessed using univariable and a linear regression model. The log₁₀ TBC for raw milk differed significantly (P < 0.05) amongst the schemes with the lowest (5.6 ± 4.7 log₁₀ cfu/ml) and highest (6.7 ± 5.8 log₁₀ cfu/ml) recorded from Marirangwe and Nharira respectively. The mean log₁₀ of TBC of processed milk (6.6 ± 6.0 log₁₀ cfu/ml) were marginally higher than those of raw milk (6.4 ± 5.6 log₁₀ cfu/ml) but not significant (P > 0.05). The coliform, E. coli and S. aureus counts for raw milk significantly differed (P < 0.05) amongst the study areas. The variation in TBC, coliforms, E. coli and S. aureus counts amongst the schemes could be attributed to differences in milking hygiene where farms with more access to training and monitoring of microbiological quality of milk had lower counts. Linear regression analysis revealed dairy scheme, delivery time and season of milking as independently associated with increased TBC of raw milk. The high TBC of raw and processed milk generally indicated low levels of milking hygienic practices, and high level of post-processing contamination, respectively. The high TBC, coliform, E. coli and S. aureus counts of both raw and processed milk may present a public health hazard. Thus, educating the farmers on general hygienic practices, quickening the delivery of milk to collection centres, or availing cooling facilities on-farm will improve the microbiological quality and safety of milk.     

Microbiological quality and potential public health risks of export meat from springbok (Antidorcas marsupialis) in Namibia

To assess the microbiological quality and safety of export game meat; i) a total of 80 pooled meat samples for aerobic plate count (APC) and Enterobacteriaceae ii) water used in harvesting and processing for microbiological quality and iii) meat and rectal contents for Salmonella spp. and Shiga toxin Escherichia coli (STEC) were evaluated in 2009 and 2010. No differences (p > 0.05) in the APCs were observed between the years, but the mean Enterobacteriaceae count for 2009 was 1.33 ± 0.69 log₁₀ cfu/cm² compared to 2.93 ± 1.50 log₁₀ cfu/cm² for 2010. Insignificant Heterotrophic Plate Count (HPC) levels were detected in 9/23 field water samples, while fecal bacterial (coliforms, Clostridium perfringens and enterococci) were absent in all samples. No Salmonella spp. was isolated and all E. coli isolates from meat were negative for STEC virulence genes (stx1, stx2, eae and hlyA), suggesting a negligible role by springbok in the epidemiology of STEC and Salmonella.     

In vitro inactivation of Escherichia coli, Staphylococcus aureus and Salmonella spp. using slightly acidic electrolyzed water

In the current study, the effectiveness of slightly acidic electrolyzed water (SAEW) on an in vitro inactivation of Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and Salmonella spp. was evaluated and compared with other sanitizers. SAEW (pH 5.6, 23 mg/l available chlorine concentration; ACC; and 940 mV oxidation reduction potential; ORP) was generated by electrolysis of dilute solution of HCl (2%) in a chamber of a non-membrane electrolytic cell. One milliliter of bacteria suspension (ca. 10–11 log₁₀CFU/ml) was mixed with 9 ml of SAEW, strong acidic electrolyzed water (StAEW; ca. 50 mg/l ACC), sodium hypochlorite solution (NaOCl; ca.120 mg/l ACC) and distilled water (DW) as control and treated for 60 s. SAEW effectively reduced the population of E. coli, S. aureus and Salmonella spp. by 5.1, 4.8, and 5.2 log₁₀CFU/ml. Although, ACC of SAEW was more than 5 times lower than that of NaOCl solution, they showed no significant bactericidal difference (p > 0.05). However, the bactericidal effect of StAEW was significantly higher (p < 0.05) than SAEW and NaOCl solution in all cases. When tested with each individual test solution, E. coli, S. aureus and Salmonella spp. reductions were not significantly different (p > 0.05). These findings indicate that SAEW with low available chlorine concentration can equally inactivate E. coli, S. aureus and Salmonella spp. as NaOCl solution and therefore SAEW shows a high potential of application in agriculture and food industry as an environmentally friendly disinfection agent.     

Efficacy of Chlorine Dioxide, Ozone, and Thyme Essential Oil or a Sequential Washing in Killing Escherichia coli O157:H7 on Lettuce and Baby Carrots

Chlorine dioxide (ClO₂), ozone, and thyme essential oil has been found to be effective in reducing pathogens, including Escherichia coli O157:H7, on selected produce. The efficacy of these sanitizers was evaluated, alone or through their sequential washing to achieve a 3 or more log reduction of mixed strains of E. coli O157:H7 on shredded lettuce and baby carrots. Samples sprinkle inoculated with mixed strains of E. coli O157:H7 were air-dried for 1 h at 22±2°C in a biosafety cabinet, stored at 4°C for 24 h, and then treated with different concentrations of disinfectants and exposure time. Sterile deionized water washing resulted in approximately 1log reduction ofE. coli O157:H7 after 10 min washing of lettuce and baby carrots. Gaseous treatments resulted in higher log reductions in comparison to aqueous washing. However, decolorization of lettuce leaves was observed during long exposure time. A logarithmic reduction of 1.48-1.97log₁₀ cfu/g was obtained using aqueous ClO₂ (10.0 mg/L for 10 min) ozonated water (9.7 mg/L for 10 min) or thyme oil suspension (1.0 mL/L for 5 min) on lettuce and baby carrots. Of the three sequential washing treatments used in this study, thyme oil followed by aqueous ClO₂/ozonated water, or ozonated water/aqueous ClO₂ were significantly (P<0.05) more effective in reducing E. coli O157:H7 (3.75 and 3.99log, and 3.83 and 4.34 log reduction) on lettuce and baby carrots, respectively. The results obtained from this study indicate that sequential washing treatments could achieve 3-4log reduction of E. coli O157:H7 on shredded lettuce and baby carrots.     

Effect of mild heat pre-treatment with alkaline electrolyzed water on the efficacy of acidic electrolyzed water against Escherichia coli O157:H7 and Salmonella on Lettuce

Cut lettuce dip-inoculated with Escherichia coli O157:H7 and Salmonella was treated with alkaline electrolyzed water (AlEW) at 20°C for 5 min, and subsequently washed with acidic electrolyzed water (AcEW) at 20°C for 5 min. Pre-treatment with AlEW resulted in an approximate 1.8 log₁₀ cfu/g reduction of microbial populations, which was significantly (p⩽0.05) greater than microbial reductions resulting from other pre-treatment solutions, including distilled water and AcEW. Repeated AcEW treatment did not show a significant bacterial reduction. Mildly heated (50°C) sanitizers were compared with normal (20°C) or chilled (4°C) sanitizers for their bactericidal effect. Mildly heated AcEW and chlorinated water (200 ppm free available chlorine) with a treatment period of 1 or 5 min produced equal reductions of pathogenic bacteria of 3 log₁₀ and 4 log₁₀ cfu/g, respectively. The procedure of treating with mildly heated AlEW for 5 min, and subsequent washing with chilled (4°C) AcEW for period of 1 or 5 min resulted in 3–4 log₁₀ cfu/g reductions of both the pathogenic bacterial counts on lettuce. Extending the mild heat pre-treatment time increased the bactericidal effect more than that observed from the subsequent washing time with chilled AcEW. The appearance of the mildly heated lettuce was not deteriorated after the treatment. In this study, we have illustrated the efficacious application of AlEW as a pre-wash agent, and the effective combined use of AlEW and AcEW.     

Adaptation of Escherichia coli O157:H7 to acid in traditional and commercial goat milk amasi

Acid resistance of Escherichia coli O157:H7 strains UT 10 and UT 15 were determined in traditional Amasi fermented for 3 days at ambient temperature (ca 30 °C) and commercial Amasi fermented at 30 °C for 24 h and stored at 7 °C for 2 days. Escherichia coli O157:H7 counts in commercial Amasi were detected at 2.7 log₁₀ cfu/ml after 3 days while those in traditional Amasi could not be detected after the same period. There was no significant difference (p ? 0.05) in the survival of acid adapted (AA) and non-adapted (NA) E. coli O157:H7 in traditional Amasi, while in commercial Amasi, the NA strain survived significantly (p ? 0.05) better than its AA counterpart. Regardless of prior adaptation to acid, E. coli O157:H7 can survive during fermentation and storage of fermented goat milk Amasi. Also, the fermentation time, pH and storage temperature affects the survival of E. coli O157:H7 in the fermented milk.     

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.     

Microbiological quality of fresh lettuce from organic and conventional production

Previously there was no available information on the levels of indicator bacteria and the prevalence of pathogens in fresh lettuce grown in organic and conventional farms in Spain. A total of 72 lettuce samples (18 farms for 4 repetitions each) for each type of the agriculture were examined in order to assess the bacteriological quality of the lettuces, in particular the prevalence of selected pathogens. The lettuce samples were analyzed for the presence of aerobic mesophilic, psychrotrophic microorganisms, yeasts and moulds, Enterobacteriaceae, mesophilic lactic acid bacteria, Pseudomonas spp. and presumptive Escherichia coli, Salmonella spp. and Listeria monocytogenes. The mean aerobic mesophilic counts (AM) were 6.35 ± 0.69 log₁₀ cfu g⁻¹ and 5.67 ± 0.80 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. The mean counts of psychrotrophic microorganisms were 5.82 ± 1.01 log₁₀ cfu g⁻¹ and 5.41 ± 0.92 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. Yeasts and moulds (YM) mean counts were 4.74 ± 0.83 log₁₀ cfu g⁻¹ and 4.21 ± 0.96 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. Lactic acid bacteria (LAB) were present in low numbers and the mean counts were 2.41 ± 1.10 log₁₀ cfu g⁻¹ and 1.99 ± 0.91 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. Pseudomonas spp. mean counts were 5.49 ± 1.37 log₁₀ cfu g⁻¹ and 4.98 ± 1.26 log₁₀ cfu g⁻¹ in organic and conventional lettuce, respectively. The mean counts for Enterobacteriaceae were 5.16 ± 1.01 log₁₀ cfu g⁻¹ and 3.80 ± 1.53 log₁₀ cfu g⁻¹ in organic and conventional lettuce, respectively. E. coli was detected in 22.2% (16 samples) of organic lettuce and in 12.5% (9 samples) of conventional lettuce. None of the lettuce samples was positive for E. coli O157:H7, L. monocytogenes and Salmonella spp. From the samples analyzed by principal component analysis (PCA) a pattern with two different groups (conventional and organic) can be observed, being the highest difference between both kinds of samples the Enterobacteriaceae count.     

Variation of microbial load and visual quality of ready-to-eat salads by vegetable type,season, processor and retailer

Microbial components and visual quality were determined on 1158 consumer units of ready-to-eat salads from several processors, two per each of 579 process lots, with residual shelf-life varying around a mode of five days, collected over 19 months in the years 2006–2008 from retail stores of two Italian cities close to a major producing and processing area. The salads were mainly baby leaf of single species (lettuce, arugula, spinach, lamb’s lettuce), with approximately 10% of the lots made up by mixes of 2–4 species. One unit per lot was analyzed on the day of collection and the other at the consume-by date. No Salmonella or Listeria monocytogenes was found (detection limit: presence in 25 g). Escherichia coli was detected in 27% of the lots (detection limit: 5 cfu/g), with probability of occurrence and counts highest in Autumn and for lettuce and arugula. Average visual quality was higher and other components of the microbial load were lower in Winter and Spring compared to Summer and Autumn (−0.6 log cfu/g of total aerobic counts, −1.3 log cfu/g of coliforms, −0.6 log cfu/g of yeasts and moulds). Lactic acid bacteria were detected more frequently in Spring and Summer (up to 50% of the lots). The rate of increase of microbial populations during shelf life was not affected by the level of initial contamination. At the consume-by date total aerobic count exceeded 7.2 log cfu/g for 50% of the lots and 7.7 log cfu/g for 25%. Salads from the biggest processor and retailer showed slightly higher visual quality scores, lower odds of E. coli occurrence and lower microbial loads. Visual quality scores showed significant negative relationships with the levels of lactic acid bacteria, coliforms and total viable counts.     

A mathematical risk model for Escherichia coli O157:H7 cross-contamination of lettuce during processing

A stochastic simulation modelling approach was taken to determine the extent of Escherichia coli O157:H7 contamination in fresh-cut bagged lettuce leaving the processing plant. A probabilistic model was constructed in Excel to account for E. coli O157:H7 cross contamination when contaminated lettuce enters the processing line. Simulation of the model was performed using @Risk Palisade© Software, providing an estimate of concentration and prevalence in the final bags of product. Three different scenarios, named S1, S2, and S3, were considered to represent the initial concentration on the contaminated batch entering the processing line which corresponded to 0.01, 1 and 100 cfu/g, respectively. The model was satisfactorily validated based on Standard Error of Prediction (SEP), which ranged from 0.00-35%. ANOVA analysis performed on simulated data revealed that the initial concentration in the contaminated batch (i.e., S1, S2, and S3) did not influence significantly (p = 0.4) the E. coli O157:H7 levels in bags derived from cross contamination. In addition, significantly different (p < 0.001) prevalence was observed at the different levels simulated (S1; S2 and S3). At the lowest contamination level (0.01 cfu/g), bags were cross-contaminated sporadically, resulting in very low E. coli O157:H7 populations (mean: ≤2 cfu/bag) and prevalence levels (<1%). In contrast, higher average prevalence levels were obtained for S2 and S3 corresponding to 3.05 and 13.39%, respectively. Furthermore, the impact of different interventions on E. coli O157:H7 cross-contamination (e.g., pathogen testing, chlorination, irradiation, and cleaning and disinfection procedures) was evaluated. Model showed that the pathogen was able to survive and be present in the final bags in all simulated interventions scenarios although irradiation (0.5 KGy) was a more effective decontamination step in reducing prevalence than chlorination or pathogen testing under the same simulated conditions.     

Mixed species biofilms of Listeria monocytogenes and Lactobacillus plantarum show enhanced resistance to benzalkonium chloride and peracetic acid

We investigated the formation of single and mixed species biofilms of Listeria monocytogenes strains EGD-e and LR-991, with Lactobacillus plantarum WCFS1 as secondary species, and their resistance to the disinfectants benzalkonium chloride and peracetic acid. Modulation of growth, biofilm formation, and biofilm composition was achieved by addition of manganese sulfate and/or glucose to the BHI medium. Composition analyses of the mixed species biofilms using plate counts and fluorescence microscopy with dual fluorophores showed that mixed species biofilms were formed in BHI (total count, 8-9 log₁₀ cfu/well) and that they contained 1-2 log₁₀ cfu/well more L. monocytogenes than L. plantarum cells. Addition of manganese sulfate resulted in equal numbers of both species (total count, 8 log₁₀ cfu/well) in the mixed species biofilm, while manganese sulfate in combination with glucose, resulted in 1-2 log₁₀ more L. plantarum than L. monocytogenes cells (total count, 9 log₁₀ cfu/well). Corresponding single species biofilms of L. monocytogenes and L. plantarum contained up to 9 log₁₀ cfu/well. Subsequent disinfection treatments showed mixed species biofilms to be more resistant to treatments with the selected disinfectants. In BHI with additional manganese sulfate, both L. monocytogenes strains and L. plantarum grown in the mixed species biofilm showed less than 2 log₁₀ cfu/well inactivation after exposure for 15 min to 100 μg/ml benzalkonium chloride, while single species biofilms of both L. monocytogenes strains showed 4.5 log₁₀ cfu/well inactivation and single species biofilms of L. plantarum showed 3.3 log₁₀ cfu/well inactivation. Our results indicate that L. monocytogenes and L. plantarum mixed species biofilms can be more resistant to disinfection treatments than single species biofilms.     

Microbial safety considerations of flooding in primary production of leafy greens: A case study

This study evaluated the effects of a flood event, floodplain and climatic parameters on microbial contamination of leafy greens grown in the floodplains. Additionally, correlations between pathogenic bacteria and levels of indicator microorganisms have been also determined. To diagnose the microbial contamination after the flood event, sampling was carried out in weeks 1, 3, 5 and 7 after the flooding in four flooded lettuce fields. To assess the impact of flooding on the microbial contamination of leafy greens, indicator microorganisms (coliforms, Escherichia coli and Enterococcus) and pathogenic microorganisms (Salmonella spp., VTEC (E. coli O157:H7 and other verocytotoxin producing E. coli, O26, O103, O111, O145) and Listeria monocytogenes) were evaluated. Irrigation water, soil and lettuce samples showed levels of coliforms and E. coli higher than 5 and 3 log cfu/g or 100 mL, respectively when sampled 1 week after flooding. However, bacterial counts drastically declined three weeks after the flooding. Climatic conditions after flooding, particularly the solar radiation (6–8 MJ/m²), affected the survival of bacteria in the field. L. monocytogenes was not detected in lettuce samples, except for 2 samples collected 3 weeks after the flooding. The presence of Salmonella was detected in irrigation water, soil and lettuce by multiplex PCR one week after the flooding, but only 2 samples of soil and 1 sample of water were confirmed by colony isolation. Verotoxigenic E. coli was detected in soil and lettuce samples by multiplex PCR. Therefore, the implication of flood water as the source of VTEC contamination of soil and lettuce was not clear. E. coli counts in irrigation water were positively correlated with those in lettuce. A significant correlation (P < 0.005) was found between the presence of pathogens and E. coli counts, highlighting a higher probability of detection of pathogens when high levels of E. coli are found. The results obtained in the present study confirm previous knowledge which defined flooding as a main risk factor for the microbial contamination of leafy greens.     

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.     

Cross-contamination of fresh-cut lettuce after a short-term exposure during pre-washing cannot be controlled after subsequent washing with chlorine dioxide or sodium hypochlorite

Chlorine dioxide (ClO₂) has been postulated as an alternative to sodium hypochlorite (NaClO) for fresh-cut produce sanitization to avoid risks associated with chlorination by-products. Experiments were performed to determine the prevention of cross-contamination of fresh-cut lettuce by Escherichia coli using chlorine dioxide (3 mg/L) or sodium hypochlorite (100 mg/L) as sanitation agents. The efficacy of these sanitation solutions was evaluated simulating as much as possible the conditions of a fresh-cut processing line. Thus, to evaluate the potential risk of cross-contamination during pre-washing, inoculated fresh-cut lettuce was pre-washed and after that non-inoculated lettuce was then pre-washed in the same water. After this pre-washing, non-inoculated lettuce was cross-contaminated, changing from 0 to 3.4 log units of E. coli cells. During washing with sanitizers, none of the tested sanitation agents significantly reduced E. coli counts in both inoculated and cross-contaminated lettuce. These results suggest that when cross-contamination occurs, even if the event is recent, subsequent sanitation steps are inefficient for inactivating E. coli cells on the vegetable tissue. However, chlorine dioxide and sodium hypochlorite solutions were able to inactivate most E. coli cells that passed from inoculated product to wash water. Therefore, they might be able to avoid cross-contamination between clean and contaminated product during the washing step. Scanning electron microscopy micrographs indicated that bacterial cells were mainly located in clusters or tissue stomata where they might be protected, which explains the low efficacy of sodium hypochlorite and chlorine dioxide solutions observed in this study.     

Efficacy of various plant hydrosols as natural food sanitizers in reducing Escherichia coli O157:H7 and Salmonella Typhimurium on fresh cut carrots and apples

In the present study, inhibitory effects of the hydrosols of thyme, black cumin, sage, rosemary and bay leaf were investigated against Salmonella Typhimurium and Escherichia coli O157:H7 inoculated to apple and carrots (at the ratio of 5.81 and 5.81 log cfu/g for S. Typhimurium, and 5.90 and 5.70 log cfu/g for E. coli O157:H7 on to apple and carrot, respectively). After the inoculation of S. Typhimurium or E. coli O157:H7, shredded apple and carrot samples were washed with the hydrosols and sterile tap water (as control) for 0, 20, 40 and 60 min. While the sterile tap water was ineffective in reducing (P > 0.05) S. Typhimurium and E. coli O157:H7, 20 min hydrosol treatment caused a significant (P < 0.05) reduction compared to the control group. On the other hand, thyme and rosemary hydrosol treatments for 20 min produced a reduction of 1.42 and 1.33 log cfu/g respectively in the E. coli O157:H7 population on apples. Additional reductions were not always observed with increasing treatment time. Moreover, thyme hydrosol showed the highest antibacterial effect on both S. Typhimurium and E. coli O157:H7 counts. Inhibitory effect of thyme hydrosol on S. Typhimurium was higher than that for E. coli O157:H7. Bay leaf hydrosol treatments for 60 min reduced significantly (P < 0.05) E. coli O157:H7 population on apple and carrot samples. In conclusion, it was shown that plant hydrosols, especially thyme hydrosol, could be used as a convenient sanitizing agent during the washing of fresh-cut fruits and vegetables.     

Assessment of Escherichia coli O157:H7-specific bacteriophages e11/2 and e4/1c in model broth and hide environments

The efficacy of bacteriophages e11/2 and e4/1c as potential biocontrol agents for Escherichia coli O157:H7 in food applications was assessed under conditions relevant to the food chain environment. The stability of each phage was determined following exposure to varying environmental conditions (pH, temperature, water activity, and sodium chloride) and the ability of each phage to infect and reduce E. coli O157:H7 numbers under selected conditions was also examined. Both e11/2 and e4/1c significantly (p < 0.05) reduced numbers of E. coli O157:H7 when exposed to pH values ranging from pH > 4 to pH 9, temperatures from 4 °C to 37 °C, water activity values of 0.87 or 0.91 to 1.00 and NaCl concentrations of 1% to 2.5%. Subsequently, a cocktail of both phages was used (e11/2 and e4/1c) to assess reduction of E. coli O157:H7 on cattle hide pieces. This involved inoculating pieces of hide (20 × 20 cm) with E. coli O157:H7 (approximately 10⁶ cfu/cm²) which were subsequently treated with either a suspension of a phage cocktail, consisting of e11/2 and e4/1c (multiplicity of infection of 1000 and 10,000, respectively) or water or not treated. Two different investigations were carried out; immediately or 1 h after treatment application was performed in different experiments. Swab samples taken immediately after phage treatment showed no significant (p > 0.05) reduction of E. coli O157:H7 numbers compared to the water treated or untreated samples. However, an extended exposure time of 1 h following phage application revealed a significant reduction (p < 0.05) (1.5 log₁₀ cfu/cm² reduction) in E. coli O157:H7 numbers compared to the numbers recovered on samples treated with water only. These findings demonstrate the potential use of e11/2 and e4/1c phages as a biocontrol agent for E. coli O157:H7 within various stages of the food chain, including on cattle hide.     

Effect of cooking procedures of kiymali pide,a traditional Turkish fast-food,on destruction of Escherichia coli O157:H7

The objective of the present study was to obtain data about cooking time and temperature of kiymali pide in the restaurants and to investigate thermal inactivation of E. coli O157:H7 during experimental kiymali pide making. A field study was conducted in randomly selected 23 of 87 pide restaurants. Processing parameters including oven temperature, cooking period and post-cooking temperature were determined. Kiymali pide samples were prepared using ground beef filling experimentally inoculated with E. coli O157:H7 (7.6 log₁₀ CFU/g). Pide samples were cooked at a conventional oven at 180 °C for 180, 240, 270, 300 and 330 s. Results of the current study suggest that cooking kiymali pide at 180 °C for at least 330 s (5.5 min) may provide sufficient food safety assurance (≥ 6 log₁₀ CFU/g) for E. coli O157:H7.     

Application of high pressure processing to reduce verotoxigenic E. coli in two types of dry-fermented sausage

The effect of high pressure processing (HPP) on the survival of verotoxigenic Escherichia coli (VTEC) in two types of Norwegian type dry-fermented sausages was studied. Two different types of recipes for each sausage type were produced. The sausage batter was inoculated with 6.8 log₁₀ CFU/g of VTEC O103:H25. After fermentation, drying and maturation, slices of finished sausages were vacuum packed and subjected to two treatment regimes of HPP. One group was treated at 600 MPa for 10 min and another at three cycles of 600 MPa for 200 s per cycle. A generalized linear model split by recipe type showed that these two HPP treatments on standard recipe sausages reduced E. coli by 2.9 log₁₀ CFU/g and 3.3 log₁₀ CFU/g, respectively. In the recipe with higher levels of dextrose, sodium chloride and sodium nitrite E. coli reduction was 2.7 log₁₀ CFU/g in both treatments. The data show that HPP has a potential to make the sausages safer and also that the effect depends somewhat on recipe.     

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.     

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.