Distribution,stability, and protein interactions of Aflatoxin M1 in fresh cheese

Aflatoxin M₁ (AFM₁), a metabolite resulting from the hepatic metabolism of aflatoxin B₁, is a potential carcinogen that can be found in milk, cheese, and others dairy products. In this work, we aimed to elucidate the distribution and fate of AFM₁ in fresh cheese and whey during cheese manufacturing and storage as well as the level of interaction of this toxin with various milk, cheese, and whey proteins. Additionally, we analyzed the in vitro behavior of casein, α-lactalbumin, β-lactoglobulin, bovine serum albumin (BSA), lactoferrin, and mixtures thereof, with a fixed concentration of AFM₁ after covalent crosslinking. Our results show that up to 70% of the AFM₁ levels present in milk spiked with 0.5 and 1.5 μg L⁻¹ are released in whey during fresh cheese manufacturing. The whey and milk proteins with more AFM₁ molecules bound were α-lactalbumin and casein, with 88% and 81%, respectively. We also observed a substantial decrement in AFM₁ concentration during ripening that correlated inversely with plate counts of lactic acid bacteria and directly with the whey-draining process that occurs during fresh cheese maturation or storage. This knowledge may serve as a basis for interventions in the dairy industry aiming to increase the security of cheese and other dairy products.     

Survival of Listeria innocua in Minas Traditional Serro cheese during ripening

Cheese produced with raw milk can be a risk to consumer health. It is known that lactic acid bacteria present in raw milk and in natural starters can produce antimicrobial compounds against some foodborne bacteria. This work aimed to evaluate the survival of Listeria innocua in Minas Traditional Serro cheese during cheese ripening. The cheeses were inoculated with 10¹, 10² or 10³ CFU mL⁻¹ of the bacterium and were analyzed for 60 days of ripening at 30 °C. It was observed that the time and the dose of bacteria inoculated affected (p < 0.05) the survival of L. innocua. Even when the lowest dose was inoculated, at the end of the 60 days, approximately 10² CFU mL⁻¹ of L. innocua was detected in the cheese. The lactic acid bacteria present in the milk and in the natural starter were not sufficient to guarantee the absence of L. innocua in Minas Traditional Serro cheese even after 60 days of storage, as is required by Brazilian legislation.     

Effectiveness of a peracetic acid-based disinfectant against spores of Bacillus cereus under different environmental conditions

A peracetic acid based disinfectant was tested for its efficacy against spores of different Bacillus cereus -strains (DSM 318, 4312, 4313 and 4384). To determine the influence of different factors like exposure-time, temperature and presence of protein quantitative and qualitative suspension tests were performed. Spore suspensions of B. cereus were treated with various concentrations of a representative peracetic acid based disinfectant at three temperatures (10, 15 and 20 °C), with protein load and with different exposure times (5, 30 and 60 min). Temperature, level of concentration and exposure-time had a significant influence on reduction of spores of B. cereus (p < 0.05). The susceptibility of spores of different strains greatly differed. A treatment of spores of DSM 4384 with 2.0% for 30 min even at 10 °C inactivated all present spores (initial number 6.18–6.71 log CFU/ml). Spores of B. cereus strain DSM 4313 had only reductions of 0.16–0.97 log CFU/ml at same treatment conditions. The presence of inactivated bovine serum as interfering substance had no significant influence on reduction (p > 0.05).     

Microbial composition of turbid rice wine (Makgeolli) at different stages of production in a real processing line

The aim of this study was to examine for changes in microbial populations during the production of turbid rice wine (also known as Makgeolli) at different production plants and to identify critical points for the control of microbial quality. Samples from raw ingredients (water, rice and wheat flour), materials from different production stages (steamed ingredients, the base of fermentation, primary and secondary fermentation stages at different time points), and the final rice wine products (non–sterilized and sterilized) were analyzed. The microbiological content of samples was assessed by quantitative (aerobic plate counts, lactic acid bacteria, fungi, acetic acid bacteria, coliforms and Bacillus cereus) and qualitative (Escherichia coli and eight foodborne pathogens) analyses. Aerobic plate counts for rice and wheat flour were relatively low (3.1 and 1.9 log CFU/g, respectively), as were those for lactic acid bacteria (1.6 and 2.1 log CFU/g), and fungi counts (2.2 and 0.7 log CFU/g). All counts increased to 7.8–7.9 log CFU/ml in the base of fermentation after the addition of Nuruk and Koji, and these counts were maintained at 8–9 log CFU/ml during fermentation. Acetic acid bacteria were not detected in the ingredients, but were isolated from the base of fermentation (1.2–2.8 log CFU/ml). Heat sterilization reduced aerobic pate counts significantly from 8.4 to 2.1 log CFF/ml, and lactic acid bacteria, fungi and acetic acid bacteria were reduced to non-detectable or negligible levels. Isolated microorganisms were considered as autochthonous to the environment or were artificially introduced with the starter culture. B. cereus was widely distributed throughout the manufacturing process, and may have been introduced from the raw material (present in 100% rice and 41.7% wheat flour samples). B. cereus counts were not significantly affected by sterilization, suggesting that it may exist at low levels as spores in the final products. No coliforms and other pathogens were detected in any samples. The raw materials, the base of fermentation, and sterilization stage were identified as important points for the control of microbial quality during the fermentation process.     

Growth inhibition of Listeria monocytogenes by bacteriocin-producing Staphylococcus equorum SE3 in cheese models

The anti-listerial activity of Staphylococcus equorum SE3 isolated from cheese brine was tested in two different model cheese systems to ascertain its potential for use as a protective culture for smear cheese ripening. Co-cultivation of Listeria monocytogenes L129 and the antilisterial S. equorum SE3 was performed on “milk agar” or “modified milk agar” (model cheese surface systems). S. equorum inoculated at concentrations of 10⁶ cfu/cm² completely inhibited growth of L. monocytogenes inoculated at 10–500 cfu/cm² on modified milk agar within 24 h of incubation, in the negative controls L129 grew to >10⁷ cfu/cm². At a higher inoculation level, growth inhibition was still more than 7 log units after 24 h. L. monocytogenes strains of different serotypes were also inhibited. Co-cultivation of S. equorum SE3 with other smear bacteria or yeasts, however, showed no growth inhibition of these important ripening microorganisms. The antilisterial effect was not diminished on the modified milk agar when co-cultivation was performed with the added smear cheese microbiota. However, on milk agar with no adjuncts (“green cheese model”), only a slight (<1 log unit) growth inhibition of L. monocytogenes was observed. Addition of peptides or amino acids to milk agar could restore growth inhibition of listeriae at different levels.     

Determination of microbiological contamination sources during Turkish white cheese production

This study has been conducted to determine the microbiological contamination sources during production of white cheese in a local dairy plant, Bursa, Turkey. Twenty nine different control points or sample types (raw milk, pasteurized milk, milk in cheese vat, curd, moulded cheese before salting, moulded cheese after salting, cheese at cold holding and vacuum packaged cheese; samples from starter culture, rennet, calcium chloride solution, brine, cheese vat, cheese cloth, polyethylene separator sheet, milk stirrer, curd cutting knife, side pressure plate, upper pressure plate, moulded cheese cutting knife, cheese tray, packaging material used during production; workers’ hands, cold room and production room air, floor, wall, and potable water) have been examined for the enumeration of total aerobic mesophilic bacteria, staphylococci, coagulase positive staphylococci, Enterobacteriaceae, enterococci, coliforms, Escherichia coli, psychrophilic bacteria, and yeast and molds.We determined: starter culture as the possible contamination source of coagulase positive staphylococci, enterococci and psychrophilic bacteria; brine and upper pressure plate as the contamination source for staphylococci; floor and packaging material as the contamination source of psychrophilic bacteria; cheese vat, cheese cloth and curd cutting knife as the contamination source of total aerobic mesophilic bacteria; cold room and production room air as the contamination sources for yeast and molds.     

Short communication: Fate of major foodborne pathogens and Bacillus cereus spores in sterilized and non-sterilized Korean turbid rice wine (Makgeolli)

The objective of this study was to examine the fate of foodborne pathogens (Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella Typhimurium, and Staphylococcus aureus) and B. cereus spores in Korean turbid rice wine (Makgeolli). Samples of sterilized and non-sterilized turbid rice wine were inoculated with each of the vegetative bacteria or B. cereus spores at 3–4 log CFU/ml. The samples were stored at 5 °C or 22 °C, and bacterial survival was monitored over 28 days. Despite the harsh environment (alcohol content: 6–7% and pH: 3.43–3.98), long-term survival of pathogens was observed. Survival time was different depending on the type of beverage (pathogens survived longer in sterilized wine than in non-sterilized wine), cellular state (spores survived longer than vegetative cells), species (B. cereus survived longer than other species), and storage temperature (pathogens survived longer at 5 °C then at 22 °C). The number of B. cereus spores remained constant at both temperatures. The vegetative B. cereus population declined rapidly within 1 day, but then remained steady for up to 28 days (1.20–1.55 log CFU/ml in sterile wine). These results indicate that B. cereus formed spores that survived for a long time; therefore, it is possible that B. cereus may exist as spores in turbid rice wine. E. coli O157:H7, L. monocytogenes, S. Typhimurium, and S. aureus survived for up to 28, 14, 14, and 14 days, respectively, in sterilized wine at 5 °C. Thus, the health implications of the long-term survival of pathogens in alcoholic beverages should be carefully considered. The results provide new information that may be useful in predicting the potential microbiological hazards associated with turbid rice wine.     

Extracellular enzyme production and enterotoxigenic gene profiles of Bacillus cereus and Bacillus thuringiensis strains isolated from cheese in Turkey

The aim of the present study was to investigate the biochemical characteristics, extracellular enzyme production and enterotoxigenic genes contents of 6 Bacillus cereus and 22 Bacillus thuringiensis strains, isolated from 100 cheese samples in Turkey. Crystal morphologies of B. thuringiensis strains were found either spherical (n = 12) or spherical and irregular-shaped (n = 10) by phase contrast microscopy. B. cereus and B. thuringiensis strains were found to produce extracellular enzymes, respectively: gelatinase (83% and 91%), DNase (83% and 77%), lecithinase (83% and 95%), protease on skim milk agar (100% and 100%), protease on milk agar (100% and 91%), protease on casein agar (83% and 77%), xylanase (100% and 45%), and cellulase (0% and 41%), and amylase (83% and 27%). All of the strains, except for Bt-D1, hydrolyzed Tween 20 (96%), but not Tween 80 or tributyrin. Pectinolytic activity was obtained to be the least frequent (4%). PCR analysis showed that all strains contained nheA, nheB, nheC and hblD genes. The hblA and hblC genes were present in 2 and 4 of B. thuringiensis strains, respectively. The bceT gene was detected in 1 B. cereus and 9 B. thuringiensis strains. The entFM gene was detected more frequently in B. thuringiensis (82%) than in B. cereus strains (50%). To our knowledge, this is the first report about the isolation and identification of enterotoxigenic B. cereus and B. thuringiensis strains from cheese samples in Turkey.     

Inactivation of Listeria innocua in brined white cheese by a combination of nisin and heat

The objective of this study was to investigate the effect of nisin alone and in combination with heat (63 °C/5 min) on the inactivation of Listeria innocua in white cheese. Nisin was added at different concentrations (500, 1000, and 1500 IU ml⁻¹) to pasteurized milk before curd formation. The curd was soaked for 24 h in 10% solution of brine containing ca 10⁶ CFU ml⁻¹ of a cocktail mixture of three strains of L. innocua. Part of the nisin treated samples were heat treated at 63 °C/5 min. Total mesophilic count (TMC), L. innocua survivors and changes in the pH of white cheese were monitored each 2 d for a period of 12 d of storage at 4 or 10 °C. Nisin at 500 IU ml⁻¹ did not diminish TMC in white cheese compared to the control. The combination of heat and nisin (1000 or 1500 IU ml⁻¹) exhibited a bacteriostatic effect on TMC throughout the storage period at 4 or 10 °C. Nisin at 500 IU ml⁻¹ had a marginal inhibitory activity against L. innocua. However, nisin at 1000 and 1500 IU ml⁻¹l resulted in a more than 2 log₁₀ reduction in L. innocua count and the effect was more prominent at 10 °C. In comparison, the combination of nisin (1000 or1500 IU ml⁻¹) and heat treatment exhibited a synergistic inhibitory activity against L. innocua, where a complete elimination of the organism was accrued after 6 and 8 d of storage at 10 and 4 °C. Therefore, nisin and heat combination could be used as a prudent hurdle to preclude the growth of Listeria in white cheese, especially under the condition of abused refrigeration conditions.     

Detection of non-emetic and emetic Bacillus cereus by propidium monoazide multiplex PCR (PMA-mPCR) with internal amplification control

Bacillus cereus is an etiological agent of food-borne disease that can cause a type of emesis. To develop a sensitive and reliable diagnosis technique for detecting all the species of the B. cereus group, specific primers were designed to target a recently discovered part of the cereulide synthetase gene (cesB) for emetic B. cereus and 16S rRNA for non-emetic B. cereus. To detect PCR signals only from viable cells, propidium monoazide (PMA) was selected to eliminate the false-positive results. In addition, an internal amplification control (IAC) was applied to meet diagnostic multiplex PCR requirements that will prevent the occurrence of false-negative results. The inclusivity and exclusivity of the mPCR assay were estimated using a panel of 100 strains, including 2 emetic B. cereus, 77 non-emetic B. cereus and 21 non-Bacillus strains. The limit of detection (LOD) for dead B. cereus without PMA treatment in pure bacteria culture was 4.0 × 10² CFU/mL, as low as 7.5 × 10⁰ CFU/mL for viable B. cereus without PMA treatment, and 7.5 × 10¹ CFU/mL for viable B. cereus with PMA treatment. B. cereus in spiked food produce was detected specifically and sensitively at 1.0 × 10³ CFU/g which was the lowest concentration detected. This novel PMA-mPCR-IAC assay is rapid and reliable, providing an efficient diagnostic tool with promising application in monitoring food samples.     

Isolation, identification and monitoring of contaminant bacteria in Iranian Kefir type drink by 16S rDNA sequencing

Iranian Kefir type drink (IKTD) is a highly consumed, traditional Iranian, fermented milk product. To improve monitoring procedures for food safety 32 industrial Kefir type drinks from 4 brands and 8 different production dates as well as 32 samples from pasteurized milk of the same Kefir manufacturers and air of the production sites were analyzed for contaminations. 16S rDNA extraction from Kefir samples as well as 16S rDNA obtained from samples incubated on Columbia agar were analyzed using PCR/DGGE, cloning, sequencing and phylogenetic classification. Already DGGE analysis indicated contaminations including Bacillus strains. Subsequently analysis of cultured clones indicated contaminations with Bacillus sp. including Bacillus cereus, Bacillus thuringiensis and Paenibacillus sp. in 9 (28%) from all analyzed samples. Also 38% of pasteurized milk samples were contaminated with B. cereus. The average count of B. cereus was 74 ± 19 cfu/ml. B. cereus and B. thuringiensis were found as contaminant bacteria in the air of the all manufacturing sites. These results suggest that milk is one of the most important sources of contamination with Bacillus sp., especially B. cereus for Kefir products in Iran. But bacterial contamination in Kefir samples might also originate from the air of the production sites. 16S rDNA analysis accelerates monitoring strategies.     

Incidence of Listeria monocytogenes and Escherichia coli O157:H7 in two Kasar Cheese processing environments

This study was designed to determine the presences of two environmental pathogens in two dairy factories in Sakarya, Turkey. A total of 264 environmental samples, raw milk and cheese samples were taken at four different seasons. According to the results, Listeria monocytogenes or Escherichia coli O157:H7 was isolated from 26 or 2.7% of the samples collected from both factories, respectively. None of the cheese or curd samples were found to be positive for Listeria or E. coli O157:H7. However, 50% of raw milk samples contained Listeria innocua. Listeria was mostly isolated from the swap samples taken from the drains or the floors in processing or packaging areas. However, E. coli was also isolated from the swap samples taken from the workers’ hands and gloves as well as the drains and the floor. Only one raw milk sample contained E. coli O157:H7. A higher prevalence of both pathogens was observed in the summer months than in the other months.     

A Turkey survey of hygiene indicator bacteria and Yersinia enterocolitica in raw milk and cheese samples

In this research, a total of 200 dairy (raw milk, cheese) samples obtained from Ankara, were examined for the presence of Yersinia spp., total coliform and Escherichia coli. As expected, raw milk 55% (55/100) were significantly contaminated with Yersinia spp., than cheese samples 14% (14/100). Y. enterocolitica was the most commonly isolated species, and was recovered from 47.3% in raw milk 35.7% in cheese samples. The other Yersinia spp. were identified as Y. frederiksenii (31.0%, 21.4%), Y. kristensenii (12.7%), Y. intermedia (7.2%, 7.1%) and atypical Yersinia spp. (1.8%, 35.7%) in raw milk and cheese samples, respectively. All the samples of cheese examined were negative for Y. kristensenii. All Y. enterocolitica strains tested gave negative results in the autoagglutination tests and crystal violet binding test.Furthermore total coliform bacteria and E. coli were investigated in these samples. According to the analysis results, most of the samples containing Yersinia spp.were found high number of viable count cell total coliform than E. coli. Total coliform bacteria and E. coli, were detected (3.0 × 10⁴ cfu/ml and 1.5 × 10⁴ cfu/ml) in four milk samples containing Y. enterocolitica while, they were detected (2.5 × 10⁴ cfu/ml and 1.0 × 10⁴ cfu/ml) in eight milk samples containing Y. enterocolitica. In the present study, total coliform bacteria and E. coli were detected >1.1 × 10⁵ cfu/g in six cheese samples containing Y. enterocolitica and atypical Yersinia spp. The results indicate that Yersinia spp. is more likely to be isolated from foods with a high level of coliforms than from foods with low coliform counts.     

Occurrence of aflatoxin M1 in Italian cheese: Results of a survey conducted in 2010 and correlation with manufacturing, production season, milking animals, and maturation of cheese

Aflatoxin M₁ (AFM₁), a carcinogenic metabolite secreted into milk by animals fed with crops contaminated by aflatoxin B₁, can be found in dairy products because of its relative stability to treatments used to produce foodstuffs, and also to long-term storage. Maximum admissible limits of AFM₁ in milk have been set up worldwides; specific regulations regarding dairy products have also been established in some countries. Nevertheless, little and rather discordant data on the occurrence of such a contaminant in cheese and other dairy products is available, and mainly in those countries which are important producers and consumers of cheese, such as, for example, Italy. Therefore, a one-year survey was conducted by measuring AFM₁ contamination in cheese purchased on the Italian market. More than a hundred samples representing the highest variability in terms of type of cheese, origin, cheese-making process, and maturation were collected and analysed through a previously described ELISA method coupled to a very rapid, simple and solvent-free extraction. More than 83% of samples showed detectable levels of AFM₁ (>25 ng kg⁻¹); most of them were found to be contaminated at a level between 50 and 150 ng kg⁻¹. The measured AFM₁ concentration was correlated to four factors which were presumed to influence the contamination level: manufacturing, production season, milking animal, and maturation. Statistical analyses demonstrated that milking animals and manufacturing affect AFM₁ concentrations, as cheeses obtained from cows’ milk and from artisanal production are more contaminated than cheeses produced with milk belonging to other animals and in industrial contexts. The others two factors showed statistically non-significant differences between groups.     

Fate of aflatoxin M1 during production and storage of parmesan cheese

The fate of AFM₁ during production of a long maturing cheese (Parmesan cheese) was assessed. Different levels of AFM₁ contamination and of the fat/casein (F/C) ratio of milk were considered, in order to evaluate if these factors can influence the enrichment factor (EF) of AFM₁ in cheese. For this purpose, 24 cheese-makings were carried out using naturally contaminated milk at 3 different AFM₁ levels and at 2 F/C ratios. AFM₁ analysis was performed by HPLC in raw milk, cream, cauldron milk, liquid cattle rennet, whey, curd and cheese at 3, 9, 16 and 24 months of ageing. The mass balances of the cheese-making processes were close to 100%; in whey, AFM₁ concentration was about 40% less than the concentration in cauldron milk. The EF in curd was between 4.0 and 5.2, with an average value of 4.7 ± 0.4; this factor was not significantly affected by either AFM₁ contamination level or F/C ratio. During maturation, AFM₁ concentration and consequently EF increased from curd to 16 ageing months; successively, AFM₁ slightly decreased at 24 months and consequently the EF. At 3, 9, 16 months of maturation, the EF was significantly higher for cheeses prepared using milk with low F/C than those with high F/C milk; on the contrary, EF was not significantly influenced by the AFM₁ contamination level. In cheeses, EF values were between 4.7 and 6.3; from these results, the maximum admissible level for AFM₁ in Parmesan cheese should be about 0.275 μg kg⁻¹.     

Rapid detection of viable Bacillus cereus emetic and enterotoxic strains in food by coupling propidium monoazide and multiplex PCR (PMA-mPCR)

Bacillus cereus can cause emetic and diarrheal food poisoning. It is widespread in nature and therefore, considered a major foodborne pathogen. To develop a sensitive and reliable assay for detecting enterotoxin genes (nheA, entFM, hblD, cytK) and emetic toxin (ces), specific primers each targeting one individual gene were designed. Propidium monoazide (PMA) was coupled with the developed multiplex PCR (mPCR) for the detection of viable B. cereus. The inclusivity and exclusivity of the PMA-mPCR was confirmed using a panel of 44 strains including 17 emetic and 9 enterotoxic B. cereus reference strains and 18 non-target strains. The limit of detection (LOD) without PMA treatment in pure DNA was 2 pg/reaction tube. The LOD of mPCR assay in pure heat-killed dead bacteria was 4.0 × 10² CFU/mL. Also, the LOD on the viable bacteria with or without PMA treatment was similar (3.8 × 10² CFU/mL) showing that the PMA treatment did not significantly decrease sensitivity. Finally, the newly developed PMA-mPCR successfully detected 4.8 × 10³ and 3.6 × 10³ CFU/g of viable B. cereus F4810/72 (emetic) and B. cereus ATCC 12480 (enterotoxic) reference strains, respectively, in food samples. Hence, this study combines PMA and mPCR to detect viable B. cereus with a wide range of toxin detection (5 toxins). Thus, the novel PMA-mPCR assay developed in this study is a rapid and efficient diagnostic tool for the monitoring of viable B. cereus in food samples and potentially other samples via appropriate DNA extraction.     

Impact of Clostridium spp. on cheese characteristics: Microbiology,color, formation of volatile compounds and off-flavors

The impact of autochthonous and type-strains of Clostridium tyrobutyricum, Clostridium butyricum, Clostridium beijerinckii and Clostridium sporogenes on spoilage (late blowing defect, LBD), physico-chemical characteristics and volatile profile of cheese has been investigated. Five semi-hard cheeses were produced from ewe milk inoculated with 10⁴ spores/mL of five Clostridium strains and ripened for 60 d. One cheese without clostridial spores served as control. C. tyrobutyricum CECT 4011 and INIA 68 resulted potent cheese spoilers, and caused the appearance of the earliest and greatest symptoms of LBD, affecting cheese pH and color, and leading to accumulation of volatile compounds like butyric, propionic and pentanoic acids and some aldehydes, alcohols and esters associated with cheese rancid and pungent off-odors. Cheeses contaminated with C. beijerinckii INIA 63 and C. sporogenes INIA 71 showed milder and late LBD symptoms, and a volatile profile characterized by higher levels of 2-butanone, 2,3-butanedione and 2-butanol than the rest of cheeses. Despite cheese inoculated with C. butyricum CECT 361 presented a slight blown-pack at the end of ripening, it showed physico-chemical characteristics and a volatile profile similar to control cheese. The first two axes of a principal component analysis (PCA) performed for the 21 significant volatile compounds out of 38, accounting for 91% of the variability between cheeses, separated cheeses made with C. tyrobutyricum CECT 4011 and INIA 68, with severe LBD symptoms, from the rest of cheeses, and also differentiated control cheese and cheese made with C. butyricum CECT 361, from cheeses with milder LBD symptoms made with C. beijerinckii INIA 63 and C. sporogenes INIA 71.     

Behavior of Listeria monocytogenes type1 355/98 (85) in meat emulsions as affected by temperature,pH, water activity,fat and microbial preservatives

The aim of this work was to analyze the effect of temperature (10–30 °C), fat content (20–50%), sodium chloride (2.5–5.0%) and preservative concentrations: sodium nitrite (0–150 ppm), lactic acid (50–500 mM) and nisin (0–100 IU/g (international units per gram)) on the growth of Listeria monocytogenes in a meat emulsion system.Individual and simultaneous effects of the parameters were tested and the results were mathematical modeled; inhibition indexes were calculated in each case. The addition of 7.5% NaCI inhibited the growth of L. monocytogenes at 20 and 30 °C, however, at 10 °C, microbial counts reached approximately 10⁶ CFU/g. The addition of 50 mM of lactic acid to obtain a pH ≤ 5 inhibited the growth of L. monocytogenes. The combinations of lactic acid with sodium nitrite or with nisin showed an enhancement of the inhibitory effect. However, considering the low toxicity of nisin, the combination of lactic acid (50 mM) and nisin (20 IU/g) would be more acceptable in the prevention of the growth of Listeria monocytogenes.     

The antimicrobial effect of wine on Bacillus cereus in simulated gastro-intestinal conditions

This study aimed to evaluate the antimicrobial activity of wine against Bacillus cereus vegetative cells and spores. The results clearly show that wine exerts a strong inactivation effect against vegetative cells of B. cereus. The red wine tested inactivated stationary phase cultures to undetectable numbers in less than 10 s. Thus, further inactivation assays were carried out with wine diluted with water (1:4 and 1:8). Diluted wine 1:4 caused a reduction of approximately 5 log cycles on viable cell counts in 20 s. On the other hand, B. cereus spores were found to be highly resistant to wine exposure. The influence of wine components (organic acids, ethanol and phenolic compounds) was investigated on vegetative cells. The wine organic acids tested exhibited a strong inactivation effect, and, when combined with ethanol, a slight synergistic effect was observed. The wine phenolic compounds assayed displayed no activity against the vegetative cells at the concentrations tested. At the simulated gastric conditions studied (in the presence of food), wine diminished considerably the number of B. cereus viable cells in addition to the effect of the synthetic gastric fluid. The behaviour of B. cereus spores under gastro-intestinal conditions was also evaluated. In a consumption-like scenario, the addition of wine led to lower total counts (vegetative cells + spores) of B. cereus in the simulated intestine conditions, showing that wine inhibits the proliferation of the vegetative cells obtained from the germination of spores. This work provides evidence that consumption of wine during a meal may diminish the number of viable cells of B. cereus and reduces the impact of the germination of spores that may occur in the small intestine, thus lowering the risk of toxi-infection that may be caused by this pathogen.     

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

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