Raman spectroscopic study of electron-beam irradiated cold-smoked salmon

Raman spectroscopy studies have been carried out to determine the effect of electron-beam irradiation on cold-smoked salmon. Vacuum packed samples were electron-beam irradiated at doses between 0 and 8 kGy. The irradiation at 8 kGy originated modifications on the protein secondary structure with a decrease (p < 0.05) in α-helix and the concomitant increase (p < 0.05) in β-sheet, turns and unordered content. Irradiation (?1 kGy) provokes a decrease of 1518 cm^?1 band intensity of cold-smoked salmon indicating that the treatment could produce a decrease of carotenoid content.     

Considerations for post-lethality treatments to reduce Listeria monocytogenes from fully cooked bologna using ambient and pressurized steam

During processing of ready-to-eat (RTE) deli meats, any secondary processing procedures such as peeling and cutting introduce the distinct possibility of cross-contamination between equipment, personnel, and food. To eliminate or reduce pathogens such as Listeria monocytogenes and ensure food safety, RTE deli meats can be pasteurized prior to or after packaging. In this study, ambient steam in-package pasteurization was compared with pressurized steam prepackaging pasteurization to reduce L. monocytogenes from fully cooked RTE bologna. The bologna (14 cm diameter×1.5 cm thickness) samples were surface-inoculated to contain about 8 log₁₀ of L. monocytogenes. To achieve 2 log reductions for L. monocytogenes, the bologna samples needed to be treated for about 10 s in pressurized steam at 131 °C or for about 2.5 min in ambient steam at 100 °C. The pasteurization time using pressurized steam treatment was about 75–90% shorter than using ambient steam treatment. Pressurized steam treatment may be integrated into a vacuum packaging unit to effectively eradicate L. monocytogenes from RTE meats just prior to sealing the retail packages to further reduce the treatment time, avoid post-treatment recontaminations by pathogens, and improve food safety without detrimentally affecting meat quality.     

Relating physicochemical and microbiological safety indicators during processing of linguiça,a Portuguese traditional dry-fermented sausage

Linguiça is a Portuguese traditional fermented sausage whose microbiological quality and safety can be highly variable. In order to elucidate risk factors and the particularities of the manufacturing technology that explain the between-batch variability in total viable counts (TVC), Enterobacteriaceae, Staphylococcus aureus and Listeria monocytogenes in the product; microbiological and physicochemical characterisation of linguiça at five stages of production (i.e., raw pork meat, mixed with ingredients, macerated, smoked and ripened) was carried out. A total of six production batches were surveyed from two factories; one utilised curing salts and polyphosphate in their formulation (Factory II). The delayed fermentation in the nitrite-formulated sausages was partly responsible for the increase (p < 0.01) in Enterobacteriaceae, S. aureus and L. monocytogenes from raw meat (3.21 log CFU/g, 1.30 log CFU/g and 22.2 CFU/g, respectively) to the end of maceration (4.14 log CFU/g, 2.10 log CFU/g and 140 CFU/g, respectively) while the better acidification process in the nitrite-free sausages (Factory I) led to lower counts of S. aureus (2.64 log CFU/g) and L. monocytogenes (10 CFU/g) in the finished products. In Factory II, although L. monocytogenes entered the chain at the point of mixing, it became steadily inactivated during smoking and ripening (< 50 CFU/g), despite the initially-delayed fermentation. Nitrite had a strong effect on reducing Enterobacteriaceae throughout smoking (r = − 0.73) and ripening (r = − 0.59), while it failed to control the growth of S. aureus. The main hurdle preventing the development of S. aureus in linguiça is the pH, and other factors contributing to its control are: longer ripening days (p = 0.019), low S. aureus in raw meat (p = 0.098), properly-washed casings (p = 0.094), and less contamination during mixing (p = 0.199). In the case of L. monocytogenes, at least three hurdles hinder its development in linguiça: low a_w (p = 0.004), low pH (p = 0.040) and nitrite (p = 0.060), and other factors contributing to its control are: longer ripening (p = 0.072) and maceration (p = 0.106) periods, lower a_w at the end of smoking (p = 0.076) and properly-washed casings (p = 0.099). Results have shown that there is a need to standardise the productive process of linguiça, to optimise the initial acidification process, and to reinforce proper programmes of quality control of ingredients and good hygiene practices, so as to minimise the introduction of Enterobacteriaceae and pathogens from external sources.     

Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes in milk by caprylic acid and monocaprylin

Listeria monocytogenes and Escherichia coli O157:H7 are major foodborne pathogens implicated in various outbreaks involving pasteurized or unpasteurized milk, and various dairy products. The objective of this study was to determine the antibacterial effect of caprylic acid (CA, C_8:0) and its monoglyceride, monocaprylin (MC) on L. monocytogenes and E. coli O157:H7 in whole milk. A five-strain mixture of E. coli O157:H7 or L. monocytogenes was inoculated in autoclaved milk (10⁶ CFU/ml) containing 0, 25, or 50 mM of CA or MC. At 37°C, all the treatments, excepting 25 mm CA, reduced the population of both pathogens by approximately 5.0 log CFU/ml in 6 h. At 24 h of storage at 8°C, MC at both levels and CA at 50 mM decreased L. monocytogenes and E. coli O157:H7, respectively by >5.0 log CFU/ml. At 48 h of 4°C storage, populations of L. monocytogenes and E. coli O157:H7 were decreased to below detection level (enrichment negative) by 50 mm of MC and CA, respectively. Results indicate that MC could potentially be used to inhibit L. monocytogenes and E. coli O157:H7 in milk and dairy products, but sensory studies need to be conducted before recommending their use.     

PFGE characterisation and adhesion ability of Listeria monocytogenes isolates obtained from bovine carcasses and beef processing facilities

Listeria monocytogenes is a pathogen capable of adhering to many surfaces and forming biofilms, which may explain its persistence in food processing environments. This study aimed to genetically characterise L. monocytogenes isolates obtained from bovine carcasses and beef processing facilities and to evaluate their adhesion abilities. DNA from 29 L. monocytogenes isolates was subjected to enzymatic restriction digestion (AscI and ApaI), and two clusters were identified for serotypes 4b and 1/2a, with similarities of 48% and 68%, respectively. The adhesion ability of the isolates was tested considering: inoculum concentration, culture media, carbohydrate source, NaCl concentration, incubation temperature, and pH. Each isolate was tested at 10⁸ CFU mL^− 1 and classified according to its adhesion ability as weak (8 isolates), moderate (17) or strong (4). The isolates showed higher adhesion capability in non-diluted culture media, media at pH 7.0, incubation at 25 °C and 37 °C, and media with NaCl at 5% and 7%. No relevant differences were observed for adhesion ability with respect to the carbohydrate source. The results indicated a wide diversity of PFGE profiles of persistent L. monocytogenes isolates, without relation to their adhesion characteristics. Also, it was observed that stressing conditions did not enhance the adhesion profile of the isolates.     

Survival of Listeria monocytogenes in Galotyri,a traditional Greek soft acid-curd cheese,stored aerobically at 4°C and 12°C

Galotyri is a traditional Greek soft acid-curd cheese, which is made from ewes’ or goats’ milk and is consumed fresh. Because cheese processing may allow Listeria monocytogenes post-process contamination, this study evaluated survival of the pathogen in fresh cheese during storage. Portions (0.5 kg) of two commercial types (<2% salt) of Galotyri, one artisan (pH 4.0±0.1) and the other industrial (pH 3.8±0.1), were inoculated with ca. 3 or 7 log cfu g⁻¹ of a five-strain cocktail of L. monocytogenes and stored aerobically at 4°C and 12°C. After 3 days, average declines of pathogen's populations (PALCAM agar) were 1.3–1.6 and 3.7–4.6 log cfu g⁻¹ in cheese samples for the low and high inocula, respectively. These declines were independent (P>0.05) of the cheese type or the storage temperature. From day 3, however, declines shifted to small or minimal to result in 1.4–1.8 log cfu g⁻¹ of survivors at 28 days of storage of all cheeses at 4°C, indicating a strong “tailing” independent of initial level of contamination. Low (1.2–1.7 log cfu g⁻¹) survival of L. monocytogenes also occurred in cheeses at 12°C for 14 days, which were prone to surface yeast spoilage. When ca. 3 log cfu g⁻¹ of L. monocytogenes were inoculated in laboratory scale prepared Galotyri of pH ≅4.4 and ≅3% salt, the pathogen died off at 14 and 21 days at 12°C and 4°C, respectively, in artisan type cheeses fermented with the natural starter. In contrast, the pathogen survived for 28 days in cheeses fermented with the industrial starter. These results indicate that L. monocytogenes cannot grow but may survive during retail storage of Galotyri despite its low pH of or slightly below 4.0. Although contamination of Galotyri with L. monocytogenes may be expected low (<100 cfu g⁻¹) in practice, that long-term survival of the pathogen in commercial cheeses was shown to be unaffected by the artificial contamination level (3 or 7 logs) and the storage temperature (4°C or 12°C), which should be a concern.     

Incidence,populations and diversity of fungi from raw materials,final products and air of processing environment of multigrain whole meal bread

This study aimed to assess the incidence, to quantify and to assess the diversity of fungi in a multigrain whole meal bread processing plant. Two hundred and eight one (n = 281) samples were analyzed, including raw materials (n = 120), air samples (n = 136) and multigrain breads (n = 25). Among the raw materials, the whole corn flour showed the highest counts of fungi (4.8 log CFU/g), followed by whole-wheat flour (3.1 log CFU/g). The counts of fungi in the air of processing environment were higher in post-baking steps (oven output, cooling, slicing, packaging) than in pre-baking steps (weighing and mixer) (p < 0.05). Species of fungi isolated from spoiled bread samples stored at 5, 20, 25 and 30, and 40 °C corresponded mostly to Penicillium paneum and Penicillium polonicum isolated from 20 and 24% of samples, respectively. These species were also isolated from raw materials (P. paneum and P. polonicum) and air collected at different processing sampling points (P. polonicum). The high counts of filamentous fungi in raw materials and air samples in processing steps such as cooling, slicing, and packaging, suggest that contamination that may occur in these steps can be critical for the shelf life of breads. The results of this study highlight that the prevention of contamination of breads by fungal spores is still a challenge for bakery industries and that other strategies such as control of germination and growth of spoilage fungi through the development of more stable formulations have to be developed.     

Modeling the protective effect of aw and fat content on the high pressure resistance of Listeria monocytogenes in dry-cured ham

High pressure processing (HPP) is a promising food preservation technology as an alternative to thermal processing for microbial inactivation. The technological parameters, the type of microorganism, and the food composition can greatly affect the microbicidal potential of HPP against spoilage and pathogenic microorganisms. Presently, the number of available models quantifying the influence of food characteristics on the pathogen inactivation is scarce. The aim of this study was to model the inactivation of Listeria monocytogenes CTC1034 in dry-cured ham, as a function of pressure (347–852 MPa, 5 min/15 °C), water activity (a_w, 0.86–0.96) and fat content (10–50%) according to a Central Composite Design. The response surface methodology, based on the equation obtained with a stepwise multivariate linear regression, was used to describe the relationship between bacterial inactivation and the studied variables. According to the best fitting polynomial equation, besides pressure intensity, both a_w and fat content exerted a significant influence on HP-inactivation of L. monocytogenes. A clear linear piezoprotection trend was found lowering the a_w of the substrate within the whole range of tested pressure. Fat content was included in the model through the quadratic term and as interaction term with pressure, resulting in a particular behavior. A protective effect due to the presence of high fat content was identified for pressure treatments above ca. 700 MPa. At lower pressure, higher inactivation of L. monocytogenes occurred by increasing the fat content above 30%. The results emphasize the relevant influence of intrinsic factors on the L. monocytogenes inactivation by HPP, making necessary to assess and validate the effectiveness of HPP on specific food products and consequently set process criteria adjusted to each particular food product.     

Influence of composition and processing of Frankfurter sausages on the growth dynamics of Listeria monocytogenes under vacuum

Ready-to-eat (RTE) meat products, such as Frankfurter sausages, are often linked to cases of listeriosis, which is a disease caused by the foodborne pathogen Listeria monocytogenes. Frankfurter sausages vary, from manufacturer to manufacturer, in many aspects: (i) composition, (ii) physicochemical characteristics, (iii) background flora, (iv) processing technology and (v) organoleptic characteristics. Some of these factors have been widely investigated for their effect on L. monocytogenes on food products. However, studies on the specific effect of composition and processing technology on L. monocytogenes dynamics between two sausages are lacking. In this study, the growth dynamics of L. monocytogenes on two types of Frankfurter sausages, fresh and in brine, were studied at constant storage temperatures (4, 8 and 12 °C). Additionally, the physicochemical and compositional characteristics of both types of sausages were analysed. In order to study the isolated effect of preservatives, L. monocytogenes growth dynamics, at 4 °C and 30 °C, were studied in two types of liquid systems. These were prepared with the same level of preservatives as in the two types of sausages. Results indicated no major significant differences in physicochemical characteristics for the two types of sausages; but, statistically significant variability was detected in the concentration of preservatives. In liquid systems, the maximum specific growth rate (μ_max) remained unaffected by the effect of preservatives, but the lag phase was longer in the system mimicking fresh sausages. In sausages, the ‘in brine’ type had two-fold higher μ_max at all temperatures and shorter lag phase at 4 °C. The presence/absence of sausage skin, which was found to be impermeable from L. monocytogenes cells and was present in the fresh sausage, could explain those differences. In conclusion, this study highlighted the influence of processing factors, and specifically of the sausage casing on L. monocytogenes growth dynamics. Therefore, an edible membrane, which is heat resistant and impermeable to the cells, could be a hurdle strategy to control the microbiological food safety.     

Investigating the control of Listeria monocytogenes on alternatively-cured frankfurters using natural antimicrobial ingredients or post-lethality interventions

The objective of this study was to investigate natural antimicrobials including cranberry powder, dried vinegar and lemon juice/vinegar concentrate, and post-lethality interventions (lauric arginate, octanoic acid, thermal treatment and high hydrostatic pressure) for the control of Listeria monocytogenes on alternatively-cured frankfurters. Lauric arginate, octanoic acid, and high hydrostatic pressure (400 MPa) reduced L. monocytogenes populations by 2.28, 2.03, and 1.88 log10 CFU per g compared to the control. L. monocytogenes grew in all post-lethality intervention treatments, except after a 600 MPa high hydrostatic pressure treatment for 4 min. Cranberry powder did not inhibit the growth of L. monocytogenes, while a dried vinegar and a vinegar/lemon juice concentrate did. This study demonstrated the bactericidal properties of high hydrostatic pressure, octanoic acid and lauric arginate, and the bacteriostatic potential of natural antimicrobial ingredients such as dried vinegar and vinegar/lemon juice concentrate against L. monocytogenes.     

Effect of heat shock on thermal tolerance and susceptibility of Listeria monocytogenes to other environmental stresses

In the present study, Listeria monocytogenes Scott A, V7 and CCRC 14930 were first subjected to heat shock at 45°C for 1 h or at 48°C for 10 min. Thermal tolerance at 55°C and survival of the heat shocked as well as non-heat shocked cells of L. monocytogenes in the presence of 25% NaCl, 0.01% crystal violet, 0.1% H₂O₂, and 18% ethanol were examined.It was found that heat shock response of L. monocytogenes varied with strains, the condition of heat shock treatment and type of subsequent stress. Compared with the non-heat shocked cells, the 45°C-1 h heat shocked cells of strains Scott A and V7 showed an increased survival after exposure to 55°C for 60 min. Meanwhile, survival of the 48°C-10 min heat shocked L. monocytogenes cells, regardless of strain, exhibited no significant difference (p>0.05) with their respective control cells. Generally, heat shocking at 45°C for 1 h increased the tolerance of L. monocytogenes to NaCl, ethanol and crystal violet. On the other hand, heat shocking at 48°C for 10 min although increased the resistance of L. monocytogenes to NaCl reduced its resistance to H₂O₂ and crystal violet.     

Addition of probiotic bacteria in a semi-hard goat cheese (coalho): Survival to simulated gastrointestinal conditions and inhibitory effect against pathogenic bacteria

In this study, the survival of the probiotics Lactobacillus acidophilus (LA-5), Lactobacillus casei subsp. paracasei (L. casei 01) and Bifidobacterium lactis (BB12) incorporated in a Brazilian semi-hard goat cheese (coalho) when exposed to in vitro simulated conditions of digestion was assessed. The inhibitory effects of these probiotic bacteria were also evaluated against Listeria monocytogenes and Staphylococcus aureus in the goat coalho cheese during refrigerated storage. At the end of the in vitro digestion, all of the probiotic tested strains presented decreased (p < 0.05) viable cell counts (5.5–6.0 log cfu/g) with respect to those determined before exposure to the mouth conditions (7–8 log cfu/g). L. casei subsp. paracasei presented inhibition rate of 7.87% and 23.63% against S. aureus on the 14th and 21st day of storage at 10 °C, respectively; against L. monocytogenes these values were 12.96 and 32.99%. Positive inhibition rates of B. lactis toward S. aureus were found on the 1st, 14th and 21st days of storage (16.32%, 10.12% and 3.67%, respectively); and against L. monocytogenes only on the 1st day of storage (3.28%). From these results, goat coalho cheese could be an interesting carrier of probiotic strains of L. acidophilus, L. casei subsp. paracasei and B. lactis. Moreover, L. casei subsp. paracasei, could be used as protective culture for delaying the growth of S. aureus and L. monocytogenes in goat coalho cheese.     

High pressure carbon dioxide combined with high power ultrasound processing of dry cured ham spiked with Listeria monocytogenes

Traditionally, thermal treatments for the inactivation of Listeria monocytogenes in meat products involve undesirable changes of the product quality. In recent years, efforts have been carried out to develop alternative methods to inactivate L. monocytogenes without affecting the product quality attributes. In this context, the feasibility of combined high pressure carbon dioxide and high power ultrasound (HPCO₂ + HPU) treatment to inactivate L. monocytogenes inoculated on the surface of dry cured ham was investigated. Inactivation data were determined at 6, 8 and 12 MPa, as a function of temperature (22, 35, 45 °C) and treatment time (0.5 to 30 min), and compared to those obtained after thermal and HPCO₂ treatments.Color, pH and acidity changes of the samples after both thermal and HPCO₂ + HPU treatments were measured and compared, sensorial profile of the treated samples was evaluated by a sensory panel and shelf-life was determined by a storage study for 4 weeks at 4 °C.The results clearly revealed that HPU alone was not able to induce any microbial inactivation while HPCO₂ + HPU treatment always assured a certain level of inactivation, variable with the process temperature used: the inactivation efficiency was demonstrated higher at 35 °C rather than 22 °C and no enhancement was observed at 45 °C compared to 35 °C. Process conditions of 12 MPa, 35 °C, at 10 W for 5 min assured inactivation to undetectable level of L. monocytogenes spiked on the surface of the product with an initial concentration of about 10⁹ CFU/g. No differences were detected between acidity, pH, color and sensory attributes of the untreated and HPCO₂ + HPU treated dry cured ham surface, while slight differences were measured between the values obtained for the untreated and thermally treated samples. Additionally, the storage study demonstrated that a full microbial and quality shelf-life was assured for 4 weeks at 4 °C. The results obtained may open the doors to the application of such an innovative process at industrial level, in particular to treat ham-type or meat products.     

Antimicrobial activity of lactic acid bacteria against Listeria monocytogenes on frankfurters formulated with and without lactate/diacetate

Contamination by Listeria monocytogenes has been a constant public health threat for the ready-to-eat (RTE) meat industry due to the potential for high mortalities from listeriosis. Lactic acid bacteria (LAB) have shown protective action against various pathogenic bacteria. The aim of this study was to evaluate the antilisterial activity of a combination of three LAB strains (Lactiguard®) on L. monocytogenes. The combination of the LAB was inhibitory to L. monocytogenes inoculated onto frankfurters not containing lactate/diacetate after 8 weeks of refrigerated storage (0.6 log reduction compared to L. monocytogenes only control), and when a cell free extract (CFS) of the LAB was added with LAB even more inhibition was obtained (1.2 log reduction compared with L. monocytogenes only). In frankfurters containing lactate/diacetate the LAB and the LAB plus CFS were more effective in reducing growth of L. monocytogenes after 8 weeks of refrigerated storage (2 and 3.3 log reductions respectively).     

Genipin cross-linked antimicrobial nanocomposite films and gamma irradiation to prevent the surface growth of bacteria in fresh meats

A 125 μg/mL of nisin and 30 mM of disodium ethylenediaminetetraacetate (EDTA) were immobilized on the surface of the nanocrystal (CNC)/chitosan nanocomposite films by using genipin as a cross-linking agent. The effect of low-dose gamma irradiation on the antimicrobial activity of the films was tested in vitro against Escherichia coli and Listeria monocytogenes. The genipin cross-linked films prepared by irradiating at 1.5 kGy demonstrated the highest antimicrobial activity against both the bacteria at the end of 35 days of storage at 37 °C showing an inhibition zone of 27.1 mm for E. coli and 27.7 mm for L. monocytogenes as compared to 23.4 mm and 23.8 mm for the same respective bacteria at day 1. The films restricted the growth of psychrotrophs, mesophiles and Lactobacillus spp. (LAB) in fresh pork loin meats and increased the microbiological shelf-life of meat sample by more than 5 weeks. The films also reduced the count of E. coli and L. monocytogenes in meat samples by 4.4 and 5.7 log CFU/g, respectively, after 35 days of storage.Industrial relevanceFoodborne diseases are responsible for 9.4 million illnesses, 55,961 hospitalization and 1,391 deaths each year in the United States. In the context of a constantly growing population and globalization of markets, and the increase of the demand for ready to eat foods without synthetic additives, the development of new technologies to prevent food contamination and to reduce foodborne illnesses is important.The development of antimicrobial packaging containing natural antimicrobials has been proposed as a novel technology to assure food safety. This technology is gaining interest from researchers and industries due to its potential to prevent the surface growth of pathogenic bacteria in meat products.The limit of the use of natural polymers, in order to reduce the packaging wastes, is their high water permeability and low resistance. The use of nanocellulose can permit to reinforce film and can improve their physico-chemical properties. We have developed a novel biopolymeric matrix reinforced with a nanofiber. The nanofiber is non-toxic, natural and obtainable from renewable sources. Chitosan, is also obtained from renewable sources, non-toxic, biodegradable, has biocompatible properties, and found application in several fields including food packaging.     

Efficacy of plant essential oils as antimicrobial agents against Listeria monocytogenes in hotdogs

The purpose of this study was to evaluate the antimicrobial activity of different plant essential oils against Listeria monocytogenes, both in vitro and in a food system. Essential oils of thyme, clove, and pimenta were found to be most effective, based on disc diffusion experiments. Thyme and clove proved to be highly effective against L. monocytogenes in peptone water (1 g/l) and reduced the bacterial population below detection limits at concentrations of 1 ml/l. Experiments were also carried out in hotdogs of different fat content (zero-, low-, and full-fat) to evaluate the antimicrobial activity of essential oils against L. monocytogenes. Thyme essential oil reduced bacterial populations significantly (P⩽0.05) at 1 ml/l level in zero- and low-fat hotdogs, but not in full-fat hotdogs. At 10 ml/l level it reduced the bacterial population >1.3 log₁₀ cfu/g in zero-fat hotdogs, but was less effective in low- and full-fat hotdogs. Clove essential oil also exhibited antimicrobial activity at 1 ml/l in all hotdogs, and was more effective than thyme at 5 ml/l. However, increasing concentration to 10 ml/l did not result in significant (P⩽0.05) reduction of bacterial population. It is concluded that efficacy of essential oils was reduced in a food system due to interaction with food components.     

Effect of combinations of high-pressure treatment and bacteriocin-producing lactic acid bacteria on the survival of Listeria monocytogenes in raw milk cheese

The combined effect of high-pressure (HP) treatment and bacteriocin-producing lactic acid bacteria (BP-LAB) on the survival of Listeria monocytogenes Scott A in cheeses made from raw milk that was inoculated with the pathogen at 4.80 log cfu mL⁻¹, a commercial starter and one of seven strains of BP-LAB was investigated. On day 3, the counts of L. monocytogenes were 7.03 log cfu g⁻¹ in a control cheese (without BP-LAB, not HP treated), 6.06–6.74 log cfu g⁻¹ in cheeses with BP-LAB, 6.13 log cfu g⁻¹ in a cheese without BP-LAB and treated on day 2 at 300 MPa, 2.01 log cfu g⁻¹ in a cheese without BP-LAB and treated on day 2 at 500 MPa, 3.83–5.43 log cfu g⁻¹ in cheeses with BP-LAB and treated on day 2 at 300 MPa, and 1.81 log cfu g⁻¹ or less in cheeses with BP-LAB and treated on day 2 at 500 MPa. HP treatment was more effective on day 51 than on day 2.     

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

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

Antimicrobial activity of pediocin-producing Lactococcus lactis on Listeria monocytogenes,Staphylococcus aureus and Escherichia coli O157:H7 in cheese

The antimicrobial activity of two pediocin-producing transformants obtained from wild strains of Lactococcus lactis on the survival of Listeria monocytogenes, Staphylococcus aureus and Escherichia coli O157:H7 during cheese ripening was investigated. Cheeses were manufactured from milk inoculated with the three pathogens, each at approximately 6 log cfu mL⁻¹. Pediococcus acidilactici 347 (Ped⁺), Lc. lactis ESI 153, Lc. lactis ESI 515 (Nis⁺) and their respective pediocin-producing transformants Lc. lactis CL1 (Ped⁺) and Lc. lactis CL2 (Nis⁺, Ped⁺) were added at 1% as adjuncts to the starter culture. After 30 d, L. monocytogenes, S. aureus and E. coli O157:H7 counts were 5.30, 5.16 and 4.14 log cfu g⁻¹ in control cheese made without adjunct culture. On day 30, pediocin-producing derivatives Lc. lactis CL1 and Lc. lactis CL2 lowered L. monocytogenes counts by 2.97 and 1.64 log units, S. aureus by 0.98 and 0.40 log units, and E. coli O157:H7 by 0.84 and 1.69 log units with respect to control cheese. All cheeses made with nisin-producing LAB exhibited bacteriocin activity throughout ripening. Pediocin activity was only detected throughout the whole ripening period in cheese with Lc. lactis CL1. Because of the antimicrobial activity of pediocin PA-1, its production in situ by strains of LAB growing efficiently in milk would extend the application of this bacteriocin in cheese manufacture.