Efficacy of pulsed electric fields for the inactivation of indicator microorganisms and foodborne pathogens in liquids and raw chicken

This study investigated the ability of pulsed electric fields (PEF) to inactivate a range of microorganisms in liquid media and on raw chicken meat. The susceptibilities of ten Campylobacter isolates (seven Campylobacter jejuni isolates and three Campylobacter coli isolates), Escherichia coli (ATCC 25922) and Salmonella Enteritidis (ATCC 13076) to PEF in liquid media were investigated. All Campylobacter isolates tested in liquid were susceptible to PEF treatment (65 kV/cm, 5 μs, 500 Hz) with reductions of between 4.33 and 7.22 log₁₀ CFU/mL observed for the longest treatment (30 s). Significant differences in susceptibility were observed between Campylobacter isolates subjected to equivalent PEF treatments ranging from 2.41 to 5.19 log₁₀ CFU/mL. Campylobacter isolates processed in liquid media were found to be more sensitive to PEF than E. coli and S. Enteritidis (P < 0.05). The application of PEF (3.75 and 15 kV/cm, 10 μs, 5 Hz) to inoculated samples of raw chicken did not result in any significant reductions in total viable counts, Enterobacteriaceae, C. jejuni, E. coli or S. Enteritidis. Therefore, under the conditions used in this study, PEF technology may not be suitable as a food safety intervention measure for the control of microbial contaminants on broilers during processing although it may have potential to reduce contamination of process water (e.g. scald or spin chill water).     

Prevalence and quantification of Campylobacter contamination on raw chicken carcasses for retail sale in China

The quantitative contamination load of Campylobacter on raw chicken carcasses at retail outlets in seven provinces and cities of China was determined. A total of 1587 carcasses over 12 consecutive months were sampled. The overall Campylobacter contamination rate was 45.1% and 19.8% of contaminated carcasses was higher in the load than 3.0 log₁₀CFU/g. The median load was 2.1 log₁₀CFU/g with 1.4 log₁₀CFU/g as the 25th percentile and 2.9 log₁₀CFU/g as the 75th percentile, respectively. Using logistic regression, it observed the significant provincial and monthly variations in prevalence, freshly slaughtered chicken carcasses (63.1%) were found to be 2.3 times higher compared to chilled carcasses (32.9%), while no statistical significance was observed in prevalence among chicken carcasses that had various packaging solution (P = 0.370) nor from different market types (P = 0.680) suggesting that possible cross-contamination had occurred during processing and transportation. The present study found that 52.1%, 59.9%, 2.9%, 8.8% and 8.3% of carcasses were contaminated with C. jejuni, C. coli, C. lari, C. fetus, and C. upsaliensis, respectively. A higher proportion of C. coli (45.4%) than C. jejuni (39.5%) were cultured from the contaminated carcasses and this finding was suggestive of a high degree of cross-contamination occurred not just among poultry products, but also with pork products before/at retail points in China. This study provided quantitative data suitable for a risk assessment model designed to evaluate useful intervention methods to facilitate a reduction in the risk of campylobacteriosis arising from the consumption of contaminated domestically produced chicken meat in China.     

Derivation of performance objectives for Campylobacter in broiler carcasses taking into account impact of selected factors on pathogen prevalence and counts

Food safety standards in the European Union include microbiological criteria and targets in primary production. The current paper provides a strategy to elucidate risk-based metrics such a potential Food Safety Objective for Campylobacter used as benchmark to derive possible Performance Objectives for the pathogen in broiler carcasses tested after chilling. The Performance Objectives were developed using the EFSA data collected on broiler carcasses during the monitoring study performed in 2008 in the European Union according to the evaluation of the different risk factors included in the survey. The FSO for Campylobacter was set at −1.2 log₁₀ cfu/g (∼6 cfu/100 g). The Campylobacter concentrations after chilling resulting in a final concentration equal or below this proposed FSO were suggested as possible POs. The results obtained indicated that batches originating from previously thinned flocks can be more at risk of being colonized with Campylobacter. In fact, the estimated mean concentrations of Campylobacter on carcasses were 1.05 and 2.38 log₁₀ cfu/g for non thinned and thinned flocks, respectively. Further, the impact of high Campylobacter contamination on carcasses (>2.5 log₁₀ cfu/g) was shown since for those carcasses a reduction in PO values higher than 1.5 log₁₀ cfu/g is needed to meet the FSO. In contrast no significant differences for PO values estimated were found between slaughterhouses with different capacity and for carcasses tested at different times from collection. This study provides a validated methodology for the estimation of risk-based metrics based on a quantitative approach allowing food safety authorities to develop specific microbiological criteria.     

Removal of Listeria monocytogenes, Staphylococcus aureus and Escherichia coli O157:H7 biofilms on stainless steel using scallop shell powder

Biofilms on steel surfaces containing Listeria monocytogenes, Staphylococcus aureus and Escherichia coli O157:H7 continue to threaten dairy and meat processors. In this study, the ability of scallop shell powder (SSP) to remove biofilms formed by these three pathogens on stainless steel plates was examined. Whey powder solution (WPS) and bench wash water (BWW) provided by dairy and meat factories, respectively, were inoculated with L. monocytogenes, S. aureus or E. coli O157:H7 (9 log₁₀ CFU/ml). Stainless steel plates (10 cm²) were placed in the inoculated fluids and incubated at 20 °C at 48 h to form biofilms. After drying and washing in sterile water, the plates were treated with 0.0, 0.25, or 0.50% (w/v) SSP slurries for 1, 5, or 10 min and then quantitatively examined for the three pathogens. Both 0.25 and 0.50% SSP reduced L. monocytogenes on the plates by 4 log CFU/cm² with a 1 min exposure to 0.50% SSP decreasing S. aureus by 5 logs CFU/cm². After 1 min in 0.25 and 0.50% SSP, E. coli O157:H7 populations in WPS and BWW biofilms decreased 4 and 6 log CFU/cm² and 3 and 5 log CFU/cm², respectively. Increasing the concentration of SSP led to significantly increased efficacy against the tested pathogens (P < 0.05). In conclusion, this study showed that SSP slurries could significantly reduce the numbers of L. monocytogenes, S. aureus and E. coli O157:H7 in biofilms on stainless steel surfaces.     

Quantitative estimation of Campylobacter cross-contamination in carcasses and chicken products at an abattoir

This study aimed to investigate Campylobacter contamination in carcasses and chicken products derived from a Campylobacter-negative flock when the flock is slaughtered immediately after a Campylobacter-positive flock. The first 2 flocks slaughtered on 10 different dates were investigated at an abattoir. Eighteen of the 20 flocks tested were positive for Campylobacter. A Campylobacter-negative flock was slaughtered immediately after a Campylobacter-positive flock on only 1 of the 10 slaughter dates. In this case, Campylobacter was detected in the carcasses and chicken products originating from the Campylobacter-negative flock, and all the flaA genotypes of these isolates were identical to those present in the caecal contents, carcasses, and chicken products from the Campylobacter-positive flock. The Campylobacter concentrations in the products originating from the Campylobacter-negative flock were: close to the enumeration limit (1.7 log₁₀ cfu/carcass) in the carcass samples; and below the enumeration limit (2.0 log₁₀ cfu/g) in the liver samples. The mean Campylobacter concentrations in the carcasses and liver products originating from the 18 Campylobacter-positive flocks were 3.8 log₁₀ cfu/carcass and 2.6 log₁₀ cfu/g, respectively. While 91% (246/270) of chicken products originating from Campylobacter-positive flocks were positive for Campylobacter, chicken products originating from the remaining Campylobacter-negative flock were free from Campylobacter cross-contamination by slaughter prior to a Campylobacter-positive flock. These results prove that slaughtering Campylobacter-negative flocks does not introduce Campylobacter into the abattoirs and indicate that although carcasses and chicken products originating from the Campylobacter-negative flock were cross-contaminated with Campylobacter from the Campylobacter-positive flock slaughtered immediately before, the Campylobacter contamination levels were lower than those in carcasses and chicken products from Campylobacter-positive flocks. Based on these findings, the reduction of Campylobacter prevalence in broiler flocks should be taken as an effective control measure for preventing introduction of Campylobacter into abattoirs and consequently for reducing Campylobacter prevalence in chicken products in addition to the good hygienic practice at abattoirs and logistic slaughter.     

Quantification and variability of Escherichia coli and Staphylococcus aureus cross-contamination during serving and consumption of cooked thick porridge in Lungwena rural households, Malawi

The study investigated bacterial transfer to cooked thick porridge via ladles and hands during serving in 29 households in Lungwena, rural Malawi. Household stored water used for hand and ladle washing, was contaminated with Escherichia coli and Staphylococcus. aureus from hands of members of the household or from contaminated ladles used in food preparation. The results showed that hands became contaminated with E. coli and S. aureus cells in the range 0.6–3.7 and 2.2–4.3 log₁₀ CFU/cm², respectively, following washing with the contaminated water. Ladles became contaminated with 0.9–3.2 log₁₀ CFU/cm² of E. coli cells whereas contamination with S. aureus on ladles ranged between 1.9 and 4.6 log₁₀ CFU/cm². Bacterial transfer from hands to food ranged from <1 to 3.6 log₁₀ CFU/g for E. coli and 2.1 to 4.2 log₁₀ CFU/g for S. aureus. Ladle surfaces transferred from 1.3 to 3.1 and from 1.2 to 4.3 log₁₀ CFU/g of E. coli and S. aureus, respectively, on to the food. Contamination of food by hands was significantly (p < 0.05) higher than that of ladles and transfer of S. aureus was significantly (p < 0.05) higher than that of E. coli. The amount of bacteria transferred to the recipient depended on the wash water type and bacteria type. The study has demonstrated that although the traditional cooking of thick porridge inactivates S. aureus and E. coli, the porridge can be contaminated with bacteria during consumption using hands and serving on to a plate with wooden ladles.     

Systematic review-meta-analysis of the effect of chilling on Campylobacter spp. during primary processing of broilers

Carcass chilling is a critical control point for Campylobacter spp. during the primary processing of broiler chickens. Our objective was to evaluate chilling intervention research that measured the change in Campylobacter prevalence and concentration on broiler chicken carcasses during primary processing using systematic review-meta-analysis (SR-MA) methodology. Experimental and observational research published in English that investigated impacts of chilling on Campylobacter spp. during primary processing of broiler chicken carcasses were considered. Random-effects MA of air chilling resulted in heterogenous summary effect estimates (mean reduction = 0.74 log₁₀ CFU/carcass, 95% CI: 0.32–1.17, I² = 91.3%; and odds ratio = 7.42, 95% CI: 0.32–174.05, I² = 92.3%). Random-effects MA of immersion chilling with chlorine resulted in heterogenous summary effect estimates (mean reduction = 1.74 log₁₀ CFU/carcass, 95% CI: 1.32–2.16, I² = 86.4%; and odds ratio = 0.50, 95% CI: 0.20–1.28, I² = 90.6%). Effects of immersion chilling with unspecified disinfectants were also determined and varied depending on study design. The SR-MA indicated that air chilling and immersion chilling reduce Campylobacter concentrations. Due to conflicting results across studies, the estimated average effect of air chilling on Campylobacter prevalence is not informative. Immersion chilling with chlorine demonstrated a trend towards reduced Campylobacter prevalence, but this result was not significant; results should be interpreted with caution because the overall methodological soundness of included studies was low. Existing research on the effectiveness of broiler carcass chilling on Campylobacter concentration or prevalence is limited and heterogenous. Results generated herein can inform decisions makers and stakeholders on potential effective chilling interventions, and can be used to inform quantitative microbial risk assessment to estimate processing measure impacts on public health.     

Glycerol monocaprate (monocaprin) reduces contamination by Escherichia coli and Salmonella enteritidis on hard surfaces

Emulsions of glycerol monocaprate (monocaprin) kill a variety of pathogenic bacteria and viruses in suspension. In this study the microbicidal activity of monocaprin against enterobacteria was tested on contaminated hard surfaces. Surfaces were contaminated with nutrient broth or meat juice containing large numbers of Escherichia coli or Salmonella enteritidis. They were then treated with acidified monocaprin emulsions and the surviving bacteria counted. Monocaprin killed S. enteritidis in chicken meat juice on plastic cutting boards and reduced the number of viable E. coli and S. enteritidis by more than 5 log₁₀ in 2 min on a laminated plastic kitchen counter contaminated with nutrient broth. Monocaprin rapidly killed E. coli on glass, stainless steel, laminated plastic, glazed ceramic tiles and polypropylene boards. It was most effective on glass and stainless steel and more effective on dry than on wet surfaces. It was concluded that acidified monocaprin emulsions reduce contamination by pathogenic enterobacteria on hard surfaces. They may be useful as sanitizers in the home, and possibly in public places, where contaminated surfaces are a potential source of transmission of pathogens to humans. Cleaning with monocaprin emulsions may therefore be a means to improve hygiene and infection control.     

Microbiological evaluation of fresh-cut organic vegetables produced in Zambia

This study was undertaken to assess the microbiological quality of fresh-cut organic vegetables produced in Zambia. Fresh-cut organic mixed vegetables and green beans produced in Zambia were analysed for aerobic plate counts, coliforms, Enterobacteriaceae, Escherichia coli, Bacillus cereus, Clostridium perfringens, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus, and yeast and mould counts. The study included 160 samples for most of the parameters. The vegetables were grown on farms meant primarily for the export market. The vegetables were treated/washed with 150 μg ml⁻¹ chlorine solution at the processing plant prior to sampling. The aerobic plate count ranged between 3 log₁₀ and 9.7 log₁₀ CFU/g, with the highest count recorded for green beans. The largest grouping (26.1%) of vegetable samples fell between 3 and 4 log₁₀ CFU/g. Coliform counts were between 1.0 log₁₀ and 7.7 log₁₀ CFU/g. The highest incidence level was 31.4% for total coliform counts between 3 log₁₀ and 4 log₁₀ CFU/g. E. coli was only detected on mixed vegetables in the range of 0.6 log₁₀ to 3 log₁₀ CFU/g, while Enterobacteriaceae counts ranged between 1.6 log₁₀ and 9.8 log₁₀ CFU/g with the highest counts being found on green beans. The highest incidence level was of 25.8% for counts within the same range as the aerobic plate counts. Yeast and mould counts showed the highest incidence level between 5 log₁₀ and 6 log₁₀ CFU/g with an overall range between 1.5 log₁₀ and 5.6 log₁₀ CFU/g. L. monocytogenes, Salmonella spp. and S. aureus were detected in 20%, 23.1% and 83.9% of samples, respectively . C. perfringens and B. cereus were not detected in any of the samples analysed.     

Reducing microbial counts on chicken and turkey carcasses using lactic acid

A 4% concentration of lactic acid, buffered to pH = 3.7, was applied by spraying directly after the inside-outside washer of a commercial chicken and a turkey production line with the aim of reducing microbial counts on the carcasses. Slight paling of the skin and slight greying of the leaf fat was observed on the carcasses but this was considered to be commercially acceptable. Microbial reductions varied with length of storage after killing, and after treatment with acid. For both chicken and turkey, there was a highly significant difference (p < 0.001) in aerobic plate counts between control and treated carcasses up to Day 9, but no difference on Days 13 and 16. The largest difference between the aerobic plate counts on control and treated carcasses was 2.1 log₁₀ cfu/g on chicken (Day 6) and 1.1 log₁₀ cfu/g on turkey (Day 9). The prevalence of Campylobacter on turkey carcasses on the day after slaughter was reduced significantly (p < 0.001) by the application of the acid. The results indicate that the application of lactic acid offers a method of reducing microbial counts on poultry and may have implications for shelf life and food safety.     

Prevalence and counts of Campylobacter spp. in poultry meat at retail level in Estonia

Campylobacter contamination of poultry meat at retail level was studied in two surveys during the twelve-month period of 2012 in Estonia. The data from these surveys were combined and analyzed, partially together, in order to comprehensively estimate the prevalence and possible seasonality of Campylobacter in poultry and in poultry meat products in Estonia. Mostly Estonian, Lithuanian and Latvian products, representing the most typical origins of poultry products on the Estonian retail market, were sampled and analyzed in these surveys. The first survey, organized by the Estonian Veterinary and Food Board, focused on Campylobacter prevalence in poultry meat at retail level. The second survey, at the Estonian University of Life Sciences, focused on Campylobacter prevalence and counts in fresh broiler chicken meat at retail level. Additionally, broiler chicken caecal samples were collected at slaughterhouse level for the estimation of the seasonal variation of Campylobacter colonization. Caecal samples were collected weekly from a broiler chicken slaughterhouse belonging to a company representing over 95% of all commercial broiler production in Estonia. A total of 606 poultry meat samples at retail level and 380 broiler chicken caecal samples at slaughterhouse level were collected and analyzed. A total of 20.8% of the poultry meat and 39.2% of the caecal samples were found positive for Campylobacter spp. The mean number of Campylobacters in fresh broiler chicken meat in the positive samples was 3.20 log₁₀CFU/g. A distinct seasonal variation in the Campylobacter contamination of broiler chicken meat was observed, which peaked during the warm summer period.     

Effects of spray volume, type of surface tissue and inoculum level on the survival of Escherichia coli on beef sprayed with 5% lactic acid

Membrane, fat and cut muscle surfaces of beef were inoculated with Escherichia coli at numbers about 4, 1 or −1 log cfu/cm². The inoculated meat was sprayed with water or 5% lactic acid at volumes of 0.5, 0.1 or 0.02 ml/cm². Spraying with water reduced the numbers of E. coli on membrane surfaces by up to 1 log unit, but had little effect on the numbers of E. coli on fat or cut muscle surfaces. Spraying with 5% lactic acid reduced the highest numbers of E. coli on membrane surfaces by up to 4 log units; but those numbers on fat or cut muscle surfaces were reduced by ≤1.5 log unit, and the reductions declined with decreasing volumes of 5% lactic acid. With inocula of 1 log cfu/cm², spraying lactic acid in any volume reduced the numbers of E. coli on membrane or fat surfaces by about 1 log unit, and the numbers on cut muscle surfaces by between 0.8 and 0.2 log unit. E. coli were detected in enrichment cultures of samples from all surfaces inoculated with E. coli at −1 log cfu/cm² and sprayed with 5% lactic acid at 0.5 ml/cm². The findings indicate that spraying relatively heavily contaminated cuts or trimmings with 5% lactic acid at ≥0.1 ml/cm² can be expected to reduce numbers of E. coli and, presumably, associated pathogens by between 0.5 and 1 log unit. However, such a treatment is likely to be at best marginally effective for reduce the numbers of these organisms on lightly contaminated product.     

Fate of Listeria monocytogenes and Escherichia coli O157:H7 in the presence of natural background microbiota on conventional and organic lettuce

Foodborne illnesses due to the consumption of contaminated raw vegetables is a continuing food safety concern. The limited efficacy of chlorine products to disinfect in fresh-cut industries, has led to study other methods or strategies to improve the safety of processed fresh-cut products. It has been reported that the presence of competing microorganisms on the surfaces of fresh produce can contribute to the reduction of pathogens. The aim of this study was to evaluate the interactions between the natural background microbiota of shredded conventional and organic lettuce and Listeria monocytogenes and Escherichia coli O157:H7. The effect of different initial load of background microbiota (‘low’, ‘medium’ and ‘high’) was tested for its ability to reduce L. monocytogenes and E. coli O157:H7 populations on shredded lettuce during storage at 10 ± 1 °C for 8 days. After the different pre-conditioning steps in order to obtain different initial loads of microbiota, in general, we observed that varying its size had no effect on L. monocytogenes and E. coli O157:H7 survival/growth during the storage period. Only differences on the survival/growth of L. monocytogenes and E. coli O157:H7 inoculated onto organic and conventional lettuce, respectively at the end of storage period at 10 °C were found. These results highlight the necessity for corrective measures to avoid contamination of fresh-cut vegetables with foodborne pathogens.     

A risk-based approach for evaluation of hygiene performance at pig slaughter

In Denmark, the pig slaughterhouses have a daily input of pigs infected and/or contaminated with Salmonella, and the slaughter hygiene has major influence on the level of Salmonella contamination on the meat leaving the slaughterhouse. However, the relationship between the effect of improved hygiene performance and the consequential reduction of human health risk has not been estimated so far. In this study, swab samples from 2702 pig carcasses were collected, originally for other purposes, from five large Danish slaughterhouses in a period from 2005 to 2007, covering all seasons of the year. The samples were analysed quantitatively for E. coli and semi-quantitatively for Salmonella. A positive association between the number of E. coli on carcasses and the prevalence of Salmonella positive carcasses was shown. For carcasses positive for Salmonella, a positive association was also shown between the number of E. coli and the number of Salmonella on the carcass. As no biological association has been reported between faecal shedding of E. coli and presence of Salmonella, the relationship was considered to be associated with the level of faecal contamination. The positive association between E. coli and Salmonella was used as basis for developing a quantitative risk assessment model for Salmonella, using the level E. coli as model input. The model output associated the hygiene performance with a relative risk estimate of human salmonellosis. The overall objective was to develop a decision support tool that can be used to support risk-based hygiene interventions in pig slaughterhouses.     

Antibiotic resistance in Campylobacter jejuni and Campylobacter coli isolated from food in Poland

This study presents the results of investigations on the susceptibility of Campylobacter spp. strains isolated from chicken meat and giblets to fluorochinolones (ciprofloxacin), macrolides (erythromycin), tetracyclines (tetracycline) and aminoglycosides (gentamicin) andV an analysis of the molecular mechanisms of resistance to the selected antibiotics.Between January 2008 and December 2009 a total of 218 samples of chicken meat and giblets from retail trade in Poland were examined. Campylobacter bacteria were found in 143 samples, that is in 65.6% of the total number embraced by the study. Campylobacter coli was the most ubiquitous - its presence was determined in 108 samples out of 143 (75.5%), whereas Campylobacter jejuni was found in 35 of the contaminated samples (24.5%).The results obtained point to the high percentage (97.9%) of Campylobacter isolates resistant to ciprofloxacin. 92 strains (64.3%) were resistant to tetracycline, 14 (9.1%) to erythromycin and only 9 (6.3%) to gentamicin. Moreover, ten out of the 143 resistant Campylobacter strains (7.0%) were found to be resistant to at least three unrelated antibiotics.     

Effect of Hibiscus sabdariffa extract and Nigella sativa oil on the growth and aflatoxin B1 production of Aspergillus flavus and Aspergillus parasiticus strains

This study was undertaken to evaluate the inhibitory effect of Hibiscus sabdariffa calyx extract at concentrations of 5, 7.5, 10 and 12.5 g/100 ml and Nigella sativa oil at concentrations of 1, 2 and 3 ml/100 ml on the growth and aflatoxin B₁ production by Aspergillus parasiticus (CBS 921.7) and Aspergillus flavus (SQU 21) strains. The inhibition of aflatoxin B₁ production by the different concentrations of H. sabdariffa calyx ranged between 91.5-97.9% and 87.1-93.3% for A. flavus and A. parasiticus strains, respectively, whereas the inhibition by different concentrations of N. sativa oil ranged between 47.9 and 58.3% for A. flavus and 32-48% for A. parasiticus strains. The different concentrations of H. sabdariffa calyx and N. sativa oil had no significant effect on the growth of either Aspergillus species. Neither H. sabdariffa calyx nor N. sativa oil detoxified pure aqueous aflatoxin. Our results suggest that H. sabdariffa calyx and N. sativa oil extracted from seeds had metabolic effects on aflatoxin biosynthesis pathway of both Aspergillus species and can be used as an effective biocontrol and non-toxic biopreservatives in food industry against aflatoxin contamination.     

The incidence of significant foodborne pathogens in domestic refrigerators

The interior surfaces of household refrigerators are at risk of becoming contaminated with foodborne pathogens, increasing the risks of cross-contamination to other food items, including higher risk ready-to-eat foods. This study determined the incidence of a number of significant foodborne pathogens, and the general hygienic status (as estimated by total viable counts (TVCs), and total coliform counts (TCCs)) on the interior surfaces of domestic refrigerators (n = 342). Campylobacter spp., Salmonella spp. and Escherichia coli O157:H7 were not recovered from any refrigerators, but Staphylococcus aureus was recovered from 6.4%, Listeria monocytogenes and E. coli from 1.2% and Yersinia enterocolitica from 0.6% of examined refrigerators. As the recovered species can survive and grow under refrigeration or conditions of mild temperature abuse, such pathogens may transfer to (and develop to clinically significant numbers in) food in domestic fridges. Such risks are of particular concern in relation to “ready-to-eat” foods, which will not receive further bactericidal treatments (cooking) before consumption. The study estimated TVCs ranging from 2.91 log₁₀ cfu/cm² to 8.78 log₁₀ cfu/cm² and TTCs ranging from 0.045 log₁₀ cfu/cm² to 5.96 log₁₀ cfu/cm² indicating very poor standards of consumer refrigerator management and hygiene, and posing risks to consumer health. The study findings highlight the importance of adequate temperature control and thorough, regular cleaning of domestic refrigerators to ensure food safety, and of effective cooking as the last link in the domestic food service chain.     

Real-time PCR with internal amplification control for the detection of Cronobacter spp. (Enterobacter sakazakii) in food samples

Cronobacter spp. (Enterobacter sakazakii) is an opportunistic pathogen and is linked with life-threatening infections in neonates. The organism has been isolated from a wide variety of foods and environments. In this study, a Taqman real-time PCR assay incorporating an internal amplification control (IAC) was developed and evaluated for specific detection of Cronobacter spp. in foods. Previously reported macromolecular synthesis (MMS) operon sequence was selected for specificity, and 67 bacterial strains, including four strains of Cronobacter spp., were evaluated. All Cronobacter strains were successfully identified, however no cross-reactivity was observed with non-Cronobacter strains. Detection limit of the assay in pure culture and formula infant without enrichment was 1.2 × 10³ CFU/ml (1.2 × 10¹ CFU/assay). After 24 h enrichment in broth, as few as 10⁰ CFU/ml or g of Cronobacter could be detected in artificially contaminated food samples (infant formula, sterilized milk and chicken meat). The detection limit of the real-time PCR assay however remained unaffected in the presence of 10⁸ CFU/ml Salmonella typhimurium in another analysis. A total of 92 food samples were analyzed for the presence of Cronobacter, out of which two pork samples were found as positive by real-time PCR, whereas only one was detected by the ISO standard method. The adjusted real-time PCR assay can be adopted to rapidly detect Cronobacter spp. in food samples with high specificity and sensitivity, and can prevent false negative results by using the IAC.     

Inhibition of Listeria monocytogenes by pomegranate (Punica granatum) peel extract in meat paté at different temperatures

Natural antimicrobials are being more and more considered as alternative approach for controlling growth of microorganisms in food. The objective of this study was to evaluate the pomegranate extract’s (PE) potential to be used as a natural preservative in ready to eat meats. Listeria monocytogenes was the main target. In a preliminary assessment with the disk diffusion method PE showed inhibitory effect against all five tested species, in the following order of increasing sensitivity: L. monocytogenes, Bacillus subtilis, Bacillus cereus, Escherichia coli and Staphylococcus aureus.No viable cells of L. monocytogenes were detected after incubation in BHI broth in presence of 7.5% v/v of the liquid PE (or 24.7 mg dry PE/ml). This concentration was considered as the Minimal Bactericidal Concentration (MBC) of the tested PE. Two pure components commonly found in PE, namely gallic and ellagic acids were also tested in BHI broth, however they did not show considerable inhibition of L. monocytogenes. PE in a concentration equal to the measured MBC was tested against L. monocytogenes in meat paté at different temperatures. At 4 °C during 46 days the extract inhibited the growth in meat paté by 4.1 log CFU/g compared to the control, which had reached log 9.2 CFU/g already on the 18th day. Inhibition was less pronounced at higher temperatures. The results indicate that the PE has a potential to be used as a natural preservative in meat products.