Fluctuation in contamination dynamics of L. monocytogenes in quargel (acid curd cheese) lots recalled during the multinational listeriosis outbreak 2009/2010

For the first time it has been possible to determine the contamination level of Listeria monocytogenes in the very cheese lots of acid curd cheese that caused a multinational outbreak between 2009/2010. The listeriosis outbreak accounted for 34 clinical cases and eight deaths. The cheese, which was distributed in Austria, Germany, the Czech Republic, Poland and Slovakia, was recalled on the 23rd January 2010. All recalled lots were immediately investigated after call back from the retail market. The company manufactured two different cheese types, (i) red smear ripened--and (ii) mold coated/white veined--acid curd cheese. Depending on the lot production dates, cheese samples (n=1045) were analyzed at three different time points: (i) beginning to mid shelf-life (lot nos. 15-18; production period 5.1.2010-13.1.2010); (ii) end of shelf-life (lot nos. 9-18; production period 21.12.2009-13.1.2010) and, (iii) ≤46days after the expiry date (lot nos. 1-18; production period 1.12.2009-13.1.2010). Qualitative and quantitative examinations of cheese samples were performed according to ISO 11290-1&2. Examination of the samples, according to ISO 11290-1, resulted in 16 L. monocytogenes positive (red smear type) and two negative lots (mold coated type). These results were confirmed by a combined enrichment/real-time PCR method. The contamination values obtained by quantitative ISO 11290-2 varied from ≤log 2 cell forming units (CFU)/g to log 8.1CFU/g. Three out of sixteen L. monocytogenes positive lots revealed a contamination level of ≤log 2CFU/g at the beginning of their shelf-life when stored at 4°C. Nevertheless, by increasing the storage life and/or the storage temperature (15, 22°C) the contamination level could be raised to between log 5 and log 6CFU/g. Our data indicate that 81.3% (13/16) of the recalled red smear quargel cheese lots were highly contaminated with L. monocytogenes. All this implies that the main contamination of the quargel cheese took place during the red smear process and that quargel cheese can easily support growth of L. monocytogenes.     

Growth potential of Listeria monocytogenes strains in mixed ready-to-eat salads

In this study, a microbiological challenge test in three artificially contaminated retail mixed mayonnaise-based ready-to-eat salads stored at refrigerator temperatures (3 °C and 7 °C) for 48 h was carried out. Shrimp-tomato salad, smoked ham salad and garlic cheese salad were separately contaminated by a suspension of particular Listeria monocytogenes strains. The number of L. monocytogenes, Enterobacteriaceae, staphylococci and total plate count (CFU/g) was determined. Listeria monocytogenes growth potential in the salads was calculated and evaluated.A significant increase in total plate count and L. monocytogenes count throughout storage of all three investigated salads was found. Enterobacteriaceae levels were high at the beginning in all salads but significantly (p < 0.05) decreased throughout the experiment depending on the temperature.All investigated L. monocytogenes strains demonstrated growth at both temperatures but expressed different growth potential. Especially garlic cheese salad and smoked ham salad were able to support the growth of Listeria. Shrimp-tomato salad supported growth the least. The growth potential increased with the increasing temperature and exceeded 0.5 log₁₀ CFU/g in many cases. If the potential for growth is > 0.5 log₁₀ CFU/g, food products can potentially endanger human health. Reference strain (ATCC 7644) showed the least growth potential almost in all cases in comparison with strains isolated from frozen pollock loins and from thermally treated specialty sausage containing preservatives. To eliminate the occurrence of microbiological risks, the shelf-life of the studied salads was estimated.     

Rapid detection of Listeria monocytogenes in food using culture enrichment combined with real-time PCR

A rapid method for the detection of Listeria monocytogenes in foods combining culture enrichment and real-time PCR was compared to the ISO 11290-1 standard method. The culture enrichment component of the rapid method is based on the ISO standard and includes 24 h incubation in half-Fraser broth, 4 h incubation in Fraser broth followed by DNA extraction and real-time PCR detection of the ssrA gene of L. monocytogenes. An internal amplification control, which is co-amplified with the same primers as the L. monocytogenes DNA, was also included in the assay. The method has a limit of detection of 1–5 CFU/25 g food sample and can be performed in 2 working days compared to up to 7 days for the ISO standard. A variety of food samples from retail outlets and food processing plants (n = 175) and controls (n = 31) were tested using rapid and conventional methods. The rapid method was 99.44% specific, 96.15% sensitive and 99.03% accurate when compared to the standard method. This method has the potential to be used as an alternative to the standard method for food quality assurance providing rapid detection of L. monocytogenes in food.     

High prevalence,low counts and uncommon serotypes of Listeria monocytogenes in linguiça, a Brazilian fresh pork sausage

Linguiça is a highly popular and appreciated pork product in Brazil, frequently consumed undercooked. Aiming at collection of data for a future risk assessment, this study evaluated the prevalence and counts of Listeria monocytogenes in linguiça samples collected at retail level in Sao Paulo, SP, Brazil. ISO methods were used for detection and enumeration of the pathogen (11290-1 and 11290-2, respectively). Isolates were submitted to Simplex-PCR for hlyA gene and those with biochemical features of L. monocytogenes and hlyA positive were serotyped using a Multiplex PCR. Ninety percent of the samples were positive for Listeria spp., and L. monocytogenes was detected in 42% of the samples, with counts below 10² CFU/g in all samples. A prevalence of uncommon serotypes 4a and 4c was observed.     

Listeria monocytogenes and ready-to-eat seafood in Spain: Study of prevalence and temperatures at retail

The aim of this study was to obtain data from refrigerated ready-to-eat seafood products at retail in Spain (young eels, crabstick and smoked salmon), regarding prevalence and levels of Listeria monocytogenes, storage temperatures and the impact of transport conditions (type of bag) on the temperature of the product. The one-year surveillance period was carried out according to the EC Regulation No. 2073/2005, taking 5 units/batch and analyzing 250 samples following ISO 11290-1/A1 and ISO 11290-2/A methodologies. Low prevalence of L. monocytogenes was observed in surimi products, while 4.8% of smoked salmon samples were positive for Listeria with low levels (<10 cfu/g) and uneven pathogen distribution. A single company was responsible for 80% of the positive lots. All purchased products showed values higher than 4 °C at retail and an average increase of 2.5 °C or up to 6.2 °C was recorded when isothermal or plastic shopping bags were used for transport, respectively. To avoid noncompliance of the Food Safety Objective for L. monocytogenes in seafood RTE products more efforts from all stakeholders are needed, with special attention so as to improve control and maintenance of refrigerators at retail and to enhance consumer education regarding food safety practices.     

Smearing of soft cheese with Enterococcus faecium WHE 81, a multi-bacteriocin producer,against Listeria monocytogenes

Enterococcus faecium WHE 81, a multi-bacteriocin producer, was tested for its antimicrobial activity on Listeria monocytogenes in Munster cheese, a red smear soft cheese. The naturally delayed and superficial contamination of this type of cheese allowed the use of E. faecium WHE 81 at the beginning of the ripening as a surface culture. A brine solution inoculated at 10⁵ CFU of E. faecium WHE 81 per mL was sprayed on the cheese surface during the first smearing operation. On day 7, smearing of cheese samples with a brine solution at 10² CFU of L. monocytogenes per mL yielded initial cell counts of approximately 50 CFU g⁻¹ of the pathogen on the cheese surface. Although, in some instances, L. monocytogenes could survive (<50 CFU g⁻¹) in the presence of E. faecium WHE 81, it was unable to initiate growth. In control samples however, L. monocytogenes counts often exceeded 10⁴ CFU g⁻¹. In other respects, E. faecium WHE 81, which naturally existed in Munster cheese, did not adversely impact on the ripening process.     

Validation of a stochastic modelling approach for Listeria monocytogenes growth in refrigerated foods

A stochastic modelling approach was developed to describe the distribution of Listeria monocytogenes contamination in foods throughout their shelf life. This model was designed to include the main sources of variability leading to a scattering of natural contaminations observed in food portions: the variability of the initial contamination, the variability of the biological parameters such as cardinal values and growth parameters, the variability of individual cell behaviours, the variability of pH and water activity of food as well as portion size, and the variability of storage temperatures. Simulated distributions of contamination were compared to observed distributions obtained on 5 day-old and 11 day-old cheese curd surfaces artificially contaminated with between 10 and 80 stressed cells and stored at 14 °C, to a distribution observed in cold smoked salmon artificially contaminated with approximately 13 stressed cells and stored at 8 °C, and to contaminations observed in naturally contaminated batches of smoked salmon processed by 10 manufacturers and stored for 10 days a 4 °C and then for 20 days at 8 °C. The variability of simulated contaminations was close to that observed for artificially and naturally contaminated foods leading to simulated statistical distributions properly describing the observed distributions. This model seems relevant to take into consideration the natural variability of processes governing the microbial behaviour in foods and is an effective approach to assess, for instance, the probability to exceed a critical threshold during the storage of foods like the limit of 100 CFU/g in the case of L. monocytogenes.     

Loss of viability of Listeria monocytogenes in contaminated processed cheese during storage at 4, 12 and 22 °C

The behaviour of Listeria monocytogenes in a processed cheese product was evaluated over time by inoculating the product with three different L. monocytogenes strains (Scott A, CA and a strain isolated from processed cheese) at three different inoculation levels (ca. 6 × 10⁵, ca. 6 × 10³ and 10² CFU/g of cheese or less) and after storage of the contaminated products at 4, 12 or 22 °C. Growth of L. monocytogenes was not observed in any of the experimental trials (experiments involving different combinations of strain, inoculum level and storage temperature) throughout the storage period. L. monocytogenes populations decreased over time with a rate that was strain- and storage temperature-dependent. Nonetheless, for cheeses that had been inoculated with the higher inoculum and stored at 4 °C viable populations of L. monocytogenes could be detected for up to nine months post-inoculation. The L. monocytogenes survival curves obtained from the different trials were characterised by a post-inoculation phase during which the populations remained essentially unchanged (lag phase) followed by a phase of logarithmic decline. The duration of the lag phase and the rate of inactivation of L. monocytogenes in the different trials were estimated based on data from the linear descending portions of the survival curves. In addition, a non-linear Weibull-type equation was fitted to the data from each survival curve with satisfactory results. The results of the present study emphasize that, according to the definition laid down in the European Union Regulation 1441/2007, the processed cheese product tested in this work should be considered and classified as one that does not support the growth of L. monocytogenes under reasonable foreseeable conditions of distribution and storage. However, post-processing contamination of the product should be austerely avoided as the pathogen can survive in the product for extended periods of time, particularly under refrigerated storage (4 °C).     

The effect of sodium reduction with and without potassium chloride on the survival of Listeria monocytogenes in Cheddar cheese

Sodium chloride (NaCl) in cheese contributes to flavor and texture directly and by its effect on microbial and enzymatic activity. The salt-to-moisture ratio (S/M) is used to gauge if conditions for producing good-quality cheese have been met. Reductions in salt that deviate from the ideal S/M range could result in changing culture acidification profiles during cheese making. Lactococcus lactis ssp. lactis or Lc. lactis ssp. cremoris are both used as cultures in Cheddar cheese manufacture, but Lc. lactis ssp. lactis has a higher salt and pH tolerance than Lc. lactis ssp. cremoris. Both salt and pH are used to control growth and survival of Listeria monocytogenes and salts such as KCl are commonly used to replace the effects of NaCl in food when NaCl is reduced. The objectives of this project were to determine the effects of sodium reduction, KCl use, and the subspecies of Lc. lactis used on L. monocytogenes survival in stirred-curd Cheddar cheese. Cheese was manufactured with either Lc. lactis ssp. lactis or Lc. lactis ssp. cremoris. At the salting step, curd was divided and salted with a concentration targeted to produce a final cheese with 600 mg of sodium/100 g (control), 25% reduced sodium (450 mg of sodium/100 g; both with and without KCl), and low sodium (53% sodium reduction or 280 mg of sodium/100 g; both with and without KCl). Potassium chloride was added on a molar equivalent to the NaCl it replaced to maintain an equivalent S/M. Cheese was inoculated with a 5-strain cocktail of L. monocytogenes at different times during aging to simulate postprocessing contamination, and counts were monitored over 27 or 50 d, depending on incubation temperature (12 or 5°C, respectively). In cheese inoculated with 4 log₁₀ cfu of L. monocytogenes/g 2 wk after manufacture, viable counts declined by more than 3 log₁₀ cfu/g in all treatments over 60 d. When inoculated with 5 log₁₀ cfu/g at 3 mo of cheese age, L. monocytogenes counts in Cheddar cheese were also reduced during storage, but by less than 1.5 log₁₀ cfu/g after 50 d. However, cheese with a 50% reduction in sodium without KCl had higher counts than full-sodium cheese at the end of 50 d of incubation at 4°C when inoculated at 3 mo. When inoculated at 8 mo postmanufacture, this trend was only observed in 50% reduced sodium with KCl, for cheese manufactured with both cultures. This enhanced survival for 50% reduced-sodium cheese was not seen when a higher incubation temperature (12°C) was used when cheese was inoculated at 3 mo of age and monitored for 27 d (no difference in treatments was observed at this incubation temperature). In the event of postprocessing contamination during later stages of ripening, L. monocytogenes was capable of survival in Cheddar cheese regardless of which culture was used, whether or not sodium had been reduced by as much as 50% from standard concentrations, or if KCl had been added to maintain the effective S/M of full-sodium Cheddar cheese.     

Foci of contamination of Listeria monocytogenes in different cheese processing plants

Listeria monocytogenes is a ubiquitous bacterium widely distributed in the environment that can cause a severe disease in humans when contaminated foods are ingested. Cheese has been implicated in sporadic cases and in outbreaks of listeriosis worldwide. Environmental contamination, in several occasions by persistent strains, has been considered an important source of finished product contamination. The objectives of this research were to (i) evaluate the presence of L. monocytogenes within the factory environments and cheeses of three processing plants, artisanal producer of raw ewe's milk cheeses (APC), small-scale industrial cheese producer (SSI) and industrial cheese producer (ICP) each producing a distinct style of cheese, all with history of contamination by L. monocytogenes (ii) and identify possible sources of contamination using different typing methods (arsenic and cadmium susceptibility, geno-serotyping, PFGE). The presence of markers specific for 3 epidemic clones (ECI–ECIII) of L. monocytogenes was also investigated. Samples were collected from raw milk (n = 179), whey (n = 3), cheese brining solution (n = 7), cheese brine sludge (n = 505), finished product (n = 3016), and environment (n = 2560) during, at least, a four-year period. Listeria monocytogenes was detected in environmental, raw milk and cheese samples, respectively, at 15.4%, 1.1% and 13.6% in APC; at 8.9%, 2.9% and 3.4% in SSI; and at 0%, 21.1% and 0.2% in ICP. Typing of isolates revealed that raw ewe's milk and the dairy plant environment are important sources of contamination, and that some strains persisted for at least four years in the environment. Although cheeses produced in the three plants investigated were never associated with any case or outbreak of listeriosis, some L. monocytogenes belonging to specific PFGE types that caused disease (including putative epidemic clone strains isolated from final products) were found in this study.     

Efficacy of bacteriophage LISTEXP100 combined with chemical antimicrobials in reducing Listeria monocytogenes in cooked turkey and roast beef

The aim of this study was to verify the effectiveness of the commercially available anti-Listeria phage preparation LISTEX^™P100 in reducing Listeria monocytogenes on ready-to-eat (RTE) roast beef and cooked turkey in the presence or absence of the chemical antimicrobials potassium lactate (PL) and sodium diacetate (SD). Sliced RTE meat cores at 4 and 10 °C were inoculated with cold-adapted L. monocytogenes to result in a surface contamination level of 10³ CFU/cm². LISTEX^TMP100 was applied at 10⁷ PFU/cm² and samples taken at regular time intervals during the RTE product's shelf life to enumerate viable L. monocytogenes. LISTEX^™P100 was effective during incubation at 4 °C with initial reductions of L. monocytogenes of 2.1 log₁₀ CFU/cm² and 1.7 log₁₀ CFU/cm², respectively, for cooked turkey and roast beef without chemical antimicrobials (there was no significant difference to the initial L. monocytogenes reductions in the presence of LISTEX^TMP100 for cooked turkey containing PL and roast beef containing SD-PL). In the samples containing no chemical antimicrobials, the presence of phage resulted in lower L. monocytogenes numbers, relative to the untreated control, of about 2 log CFU/cm² over a 28-day storage period at 4 °C. An initial L. monocytogenes cell reduction of 1.5 log₁₀ CFU/cm² and 1.7 log₁₀ CFU/cm², respectively, for cooked turkey and roast beef containing no chemical antimicrobials was achieved by the phage at 10 °C (abusive temperature). At this temperature, the L. monocytogenes cell numbers of samples treated with LISTEX™ P100 remained below those of the untreated control only during the first 14 days of the experiment for roast beef samples with and without antimicrobials. On day 28, the L. monocytogenes numbers on samples containing chemical antimicrobials and treated with LISTEX^TMP100 stored at 4 and 10 °C were 4.5 log₁₀ CFU/cm² and 7.5 log₁₀ CFU/cm², respectively, for cooked turkey, and 1.2 log₁₀ CFU/cm² and 7.2 log₁₀ CFU/cm², respectively, for roast beef. In both cooked turkey samples with and without chemical antimicrobials stored at 10 °C, the phage-treated samples had significantly lower numbers of L. monocytogenes when compared to the untreated controls throughout the 28-day storage period (P < 0.0001). For roast beef and cooked turkey containing chemical antimicrobials treated with LISTEX^TMP100 and stored at 4 °C, no more than a 2 log CFU/cm² increase of L. monocytogenes was observed throughout the stated shelf life of the product. This study shows that LISTEX^™P100 causes an initial reduction of L. monocytogenes numbers and can serve as an additional hurdle to enhance the safety of RTE meats when used in combination with chemical antimicrobials.     

Multiple-locus variable number of tandem repeat analysis (MLVA) of Listeria monocytogenes directly in food samples

Listeria monocytogenes is the etiologic agent of listeriosis responsible for severe and fatal infections in humans. Listeria contamination occurs quite often in a wide range of foods due to its ubiquitous nature. Isolates need to be characterized to a strain level for accurate diagnosis of Listeria infection, epidemiological studies, investigation of outbreaks and effective prevention and control of food-borne listeriosis. The purpose of this research was to evaluate the multiple-locus variable number of tandem repeat analysis (MLVA) for sub-typing L. monocytogenes isolates in pure cultures and in food matrices. Two multiplex PCR assays were formulated to amplify six specific loci using fluorescently-labeled primers; and the amplicons were analyzed by capillary electrophoresis. The MLVA method resulted in 34 unique DNA fingerprint patterns from 46 L. monocytogenes isolates of 10 serotypes which had 29 or 30 PFGE patterns with a single restriction enzyme and 34 AFLP patterns. The MLVA patterns of the 46 isolates remained unchanged in the presence of pre-enriched food matrices including sausage, ham, chicken, milk and lettuce. The MLVA method successfully typed L. monocytogenes strains spiked in cheese, roast beef, egg salad and vegetable samples after 48 h enrichment at the initial inoculation levels of 1-5 CFU per 25 g of food or higher. The limits of detection (typing) of the MLVA method were 10³-10⁴ CFU/mL of pre-enriched food broth when evaluated using post-spiked sausage, ham, chicken, milk and lettuce samples. The MLVA method was simple, highly discriminatory, and easy to perform with portable (numerical) results. To our knowledge, this is the first report that describes the application of the MLVA method directly to food samples and demonstrates the possibility to obtain rapid and accurate subtyping results before an isolate is obtained.     

Development of a membrane filtration method for enumeration of Listeria monocytogenes from soft cheese

A new and simple analytical method has been developed to quantify low levels (≤50 cfu g⁻¹) of Listeria monocytogenes in soft cheese. The technique allows the analysis of 1 g of cheese instead of 0·1 g or 0·01 g using the ISO 11290-2 standard method. The analysis protocol combines filtration of the decimally diluted cheese suspension through a 0·45-μm pore size cellulose ester membrane, and a culture of the filter on a Palcam agar. A tween 80–trypsin treatment used to increase filterability of cheese did not reduce L. monocytogenes counts. The tested method provided more precise results (nearer to the true value) compared to the ISO 11290-2 standard method in the enumeration of L. monocytogenes from artificially contaminated cheese. However, it improves neither repeatability nor reproducibility since the selected medium, Palcam, does not allow distinction between the different Listeria species.     

Production of wara, a West African soft cheese using lemon juice as a coagulant

As an important protein source for West African consumers, wara cheese made from the leave extract of Calotropis procera has extremely short shelf life of only 2-3 days [Adegoke, G. O., Nse, E. N., & Akanni, A. O. (1992). Effects of heat, processing time, and pH on the microflora, aflatoxin content, and storability of wara, a soft white cheese. Die Nahrung, 36(3), 259-264; Umoh, V. J., & Solomon, O. (2001). Safety assessment and critical control point of milk product and some cereal beverages in Northern Nigeria. In: Proceedings of USDA/USAID/NIGERIA international conference on food safety and security, August 1-3 (pp. 122-127). Ibadan, Nigeria: IITA; Belewu, M. A., Belewu, K. Y., & Nkwunonwo, C.C. (2005). Effect of biological and chemical preservatives on the shelflife of West African soft cheese. African Journal of Biotechnology, 4, 1076-1079; Adetunji, A. O., Alonge, D. O., & Chen, J. (Unpublished). Microbial quality of wara, a southwestern Nigerian soft cheese]. Lemon juice was used in this study as a substitute coagulant during wara manufacture in order to improve the microbial quality of wara. The cheese was manufactured from pasteurized milk inoculated with 10¹ or 10² CFU ml⁻¹ of Listeria monocytogenes. Samples of the milk or cheese were taken along the manufacturing steps and during a 5 d storage period at 15 and 28 °C in order to determine the populations of L. monocytogenes, total aerobes, Enterobacteriaceae, and psychrotrophs, as well as mold and yeast. On the 4th day of storage, portions of the un-inoculated control cheese from 28 °C were deep fried in vegetable oil, mimicking the practice of West African local cheese processors. The results showed that L. monocytogenes, at both inoculation levels, did not survive the manufacture of wara. In samples initially inoculated with 10¹ CFU ml⁻¹ of L. monocytogenes, the Enterobacteriaceae counts decreased from the initial 1.78 to 1.00 Log₁₀ CFU g⁻¹ with the addition of lemon juice, and became undetectable (<1.00 Log₁₀ CFU g⁻¹) at the curdling point as well as during the 5 d storage period at both temperatures. The total aerobic counts increased from the undetectable level on the 1st day of storage to 7.65 and 3.39 Log₁₀ CFU g⁻¹, respectively at 28 or 15 °C on the 5th day of storage. The psychrotrophic, as well as the yeast and mold counts increased from the undetectable levels on the 1st day of storage to 7.11 and 5.03 Log₁₀ CFU g⁻¹, respectively at 28 °C. At 15 °C however, the population of pyschrotrophs remained undetectable throughout the 5 d storage period whereas, the yeast and molds count increased to 3.08 Log₁₀ CFU g⁻¹ on day 3 before quickly decreasing to the undetectable levels on the 5th day of storage. A similar trend was observed in cheese made from the milk with an initial Listeria inoculation level of 10² CFU ml⁻¹. The results of this study showed that lemon juice significantly reduced the populations of the sampled microorganisms, especially the populations of Enterobacteriaceae.     

Salmonella surveillance and control at post-harvest in the Belgian pork meat chain

Salmonella remains the primary cause of reported bacterial food borne disease outbreaks in Belgium. Pork and pork products are recognized as one of the major sources of human salmonellosis. In contrast with the primary production and slaughterhouse phases of the pork meat production chain, only a few studies have focussed on the post-harvest stages. The goal of this study was to evaluate Salmonella and Escherichia coli contamination at the Belgian post-harvest stages. E. coli counts were estimated in order to evaluate the levels of faecal contamination. The results of bacteriological analysis from seven cutting plants, four meat-mincing plants and the four largest Belgian retailers were collected from official and self-monitoring controls. The prevalence of Salmonella in the cutting plants and meat-mincing plants ranged from 0% to 50%. The most frequently isolated serotype was Salmonella typhimurium. The prevalence in minced meat at retail level ranged from 0.3% to 4.3%. The levels of Salmonella contamination estimated from semi-quantitative analysis of data relating to carcasses, cuts of meat and minced meat were equal to −3.40 ± 2.04 log CFU/cm², −2.64 ± 1.76 log CFU/g and −2.35 ± 1.09 log CFU/g, respectively. The E. coli results in meat cuts and minced meat ranged from 0.21 ± 0.50 to 1.23 ± 0.89 log CFU/g and from 1.33 ± 0.58 to 2.78 ± 0.43 log CFU/g, respectively. The results showed that faecal contamination still needs to be reduced, especially in specific individual plants.     

Control of Listeriamonocytogenes on commercially-produced frankfurters prepared with and without potassium lactate and sodium diacetate and surface treated with lauric arginate using the Sprayed Lethality in Container (SLIC®) delivery method

Viability of Listeriamonocytogenes was monitored on frankfurters formulated with or without potassium lactate and sodium diacetate at a ratio of ca. 7:1 and treated with lauric arginate (LAE; 22 or 44 ppm) using the Sprayed Lethality in Container (SLIC®) delivery method. Without antimicrobials, pathogen numbers remained relatively constant at ca. 3.3 log CFU/package for ca. 30 d, but then increased to ca. 8.4 log CFU/package over 120 d. Regardless of whether or not lactate and diacetate were included, when treated with LAE, pathogen numbers decreased from ca. 3.3 log CFU/package to ca. 1.5 log CFU/package within 2 h, but then increased to 7.3 and 6.7 log CFU/package, respectively, after 120 d. When frankfurters were formulated with lactate and diacetate and treated with LAE, pathogen numbers decreased by ca. 2.0 log CFU/package within 2 h and remained relatively unchanged over the 120 d. These data confirm that LAE provides an initial lethality towards L. monocytogenes and when used in combination with reduced levels/ratio of lactate and diacetate as an ingredient for frankfurters provides inhibition throughout shelf life.     

Detection and characterization of Listeria monocytogenes in Sao Jorge (Portugal) cheese production

Listeria monocytogenes is a foodborne pathogen that can cause serious invasive disease in humans. Because human listeriosis cases have previously been linked to consumption of contaminated cheese, control of this pathogen throughout the cheese production chain is of particular concern. To understand the potential for L. monocytogenes transmission via São Jorge cheese, a Portuguese artisanal cheese variety that bears a Protected Denomination of Origin classification, 357 raw milk, curd, natural whey starter, and cheese samples representative of the production chain of this cheese were collected over one year and tested for the presence of L. monocytogenes and selected physicochemical parameters. Although neither L. monocytogenes nor other Listeria spp. were detected in whey, curd, or cheese samples, 2 of the 105 raw milk samples analyzed were positive for L. monocytogenes. These 2 raw milk isolates represented a ribotype that has previously been linked to multiple human listeriosis outbreaks and cases elsewhere, indicating the potential of these isolates to cause human listeriosis. On average, physicochemical parameters of São Jorge cheese ripened for 4 mo presented values that likely minimize the risk of L. monocytogenes outgrowth during ripening and storage (mean pH = 5.48; mean moisture = 37.79%; mean NaCl concentration = 4.73%). However, some cheese samples evaluated in this study were characterized by physicochemical parameters that may allow growth and survival of L. monocytogenes. Even though our results indicate that raw milk used for São Jorge cheese manufacture as well as finished products is rarely contaminated with L. monocytogenes, continued efforts to control the presence of this pathogen in the São Jorge cheese production chain are urged and are critical to ensure the safety of this product.     

Spray application of liquid smoke to reduce or eliminate Listeria monocytogenes surface inoculated on frankfurters

In a simulated post process contamination scenario liquid smoke was sprayed on the frankfurters after peeling, and then inoculated with Listeria monocytogenes (Lm). Samples that did not receive a liquid smoke spray remained at approximately 2 log cfu/cm² during the 48 h of storage while the levels on the liquid smoke treated frankfurters continued to decline until they were below detection level (1 cfu/100 cm²). A shelf-life study lasting 140 days indicated that liquid smoke suppressed the growth of Lm for up to 130 days. An application of 2 or 3 ml liquid smoke at packaging resulted in at least a 1 log reduction of Lm within 12 h post packaging.     

Effects of γ-irradiation on Listeria monocytogenes population,colour, texture and sensory properties of Feta cheese during cold storage

Feta, a white brine cheese, was produced and contaminated with Listeria monocytogenes. Contamination occurred either at the beginning (pre-process contamination) or at the end of Feta manufacturing (post-process contamination). In the first case the milk was contaminated with 10³ cfu/ml, and 2 months later, in the final product, the L. monocytogenes population was approximately 10⁵ cfu/g. In the second case, the brine (NaCl, 7% w/v), in which the Feta was packaged, was contaminated with 10³ cfu/ml. Contaminated Feta samples were vacuum-packaged and exposed to irradiation doses of 1.0, 2.5 and 4.7 kGy and stored at 4 °C for a month. In the pre-process contaminated samples none of the irradiation doses eliminated L. monocytogenes; however the highest dose reduced the viable population to a level which is in compliance with EC regulations. In the post-process contamination, the 2.5 kGy and 4.7 kGy doses reduced L. monocytogenes counts below the detection limit. Irradiation had no effect on the texture of Feta. Irradiation at 4.7 kGy increased Feta's redness and decreased its yellowness and lightness. Sensorial analyses showed that at the 4.7 kGy dose, the aroma profile of Feta was temporarily affected, since it was restored after 30 days of cold storage.     

Use of high-concentration-short-time chlorine dioxide gas treatments for the inactivation of Salmonella enterica spp. inoculated onto Roma tomatoes

Salmonella outbreaks have been recently linked to the consumption of fresh tomatoes. Thus, there is a need to develop systems that reduce the risk of microbial contamination to increase product shelf-life and keep fresh fruit attributes. The objectives of this study were to evaluate high-concentration-short-time chlorine dioxide gas treatments effects on Salmonella-inoculated Roma tomatoes and determine the optimal treatment conditions for microbial inactivation and shelf-life extension. Effects of ClO₂ concentration (2, 5, 8 and 10 mg/l) and exposure time (10, 30, 60, 120 and 180 s) on inoculated Roma tomatoes were studied. Salmonella enterica strains, serotype Montevideo, Javiana and Baildon, were used to experimentally inoculate the food product. After ClO₂ treatments, tomatoes were stored at room temperature for 28 days. Inherent microbial population, change in tomato color, and chlorine dioxide gas residuals were evaluated. ANOVA analysis showed that both ClO₂ concentration and treatment time were significant (p < 0.01) for Salmonella inactivation. Surviving Salmonella populations of 3.09, 2.17 and 1.16 log CFU/cm² were obtained treating tomatoes with 8 mg/l ClO₂ for 60 s, 10 mg/l ClO₂ for 120 s, and 10 mg/l for 180 s, respectively (initial Salmonella population: 6.03 ± 0.11 log CFU/cm²). The selected treatments significantly reduced background microflora (p < 0.05), while fruit color and residual contents were not significantly different (p > 0.05), as compared to the control. Results suggest the potential for high-concentration-short-time treatments ClO₂ gas as an effective pathogen inactivation technology for large-scale produce packing operations.