Antimicrobial activities of spice extracts against pathogenic and spoilage bacteria in modified atmosphere packaged fresh pork and vacuum packaged ham slices stored at 4 °C

The antimicrobial activity of 14 spice extracts against four common meat spoilage and pathogenic bacteria (Listeria monocytogenes, Escherichia coli, Pseudomonas fluorescens and Lactobacillus sake) was screened in cultured media (experiment 1). The results showed that individual extracts of clove, rosemary, cassia bark and liquorice contained strong antimicrobial activity, but the mixture of rosemary and liquorice extracts was the best inhibitor against all four types of microbes. Subsequently, mixed rosemary/liquorice extracts were spray-applied to inoculated fresh pork in modified atmosphere packaging (experiment 2) and to inoculated ham slices in vacuum packaging (experiment 3). The meat samples were stored at 4 °C over a 28-day period and microbial growth was monitored regularly. The L. monocytogenes population on fresh pork by day 28 decreased 2.9, 3.1 and 3.6 logs, the MAB decreased 2.7, 2.9 and 3.1 logs, the Pseudomonas spp. count decreased 1.6, 2.1 and 2.6 logs and the total coliform count decreased 0.6, 0.8 and 1.2 logs, corresponding to 2.5, 5.0 and 10.0 mg/ml of spray, respectively, when compared to control (P < 0.05). The number of L. monocytogenes on ham slices decreased 2.5, 2.6 and 3.0 logs, the MAB plate counts decreased 2.9, 3.0 and 3.2 logs and the LAB counts decreased 2.4, 2.6 and 2.8 logs (P < 0.05), respectively, after 28-days, by the same levels of mixed rosemary/liquorice extract treatments. The results demonstrated strong potential of mixed rosemary and liquorice as a natural preservative in fresh pork and ham products.     

Antimicrobial Activity of Vanillin and Mixtures with Cinnamon and Clove Essential Oils in Controlling <Emphasis Type="Italic">Listeria monocytogenes</Emphasis> and <Emphasis Type="Italic">Escherichia coli</Emphasis> O157:H7 in Milk

The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of vanillin against Listeria monocytogenes Scott A and Escherichia coli O157:H7 was determined in tripticase soy broth (TSB), pH 7 and 6, incubated at 35 °C/24 h and in semi-skim milk incubated at 35 °C/24 h and 7 °C/14 days. The influence of the fat content of milk on the antimicrobial activity of vanillin was tested in whole and skim milk incubated at 7 °C/14 days. Mixtures of clove and cinnamon with vanillin were also evaluated in semi skim milk incubated at 7 °C. The MICs for L. monocytogenes were 3,000 ppm in TSB (pH 7) and 2,800 ppm in TSB (pH 6). The MICs for E. coli O157:H7 were 2,800 ppm in TSB (pH 7) and 2,400 ppm in TSB (pH 6). The MBCs in TSB were 8,000 ppm for L. monocytogenes and 6,000 ppm for E. coli O157:H7. The pH values assayed did not influence significantly the MIC or MBC in TSB. The MICs in semi-skim milk for L. monocytogenes and E. coli O157:H7 were 4,000 and 3,000 ppm at 35 °C/24 h, and 2,500 and 1,000 ppm at 7 °C/7 days, respectively. The MBCs were 20,000 ppm for L. monocytogenes and 11,000 ppm for E. coli O157:H7. High incubation temperatures did not affect the MBC but increased the MIC of the vanillin in milk. This effect could be attributed to the increased membrane fluidity and to the membrane perturbing activity of vanillin at low temperatures. The fat in milk reduced significantly the antimicrobial activity of vanillin, probably due to effect protective of the fat molecules. Mixtures of clove and cinnamon leaves inhibited the growth of L. monocytogenes in a similar way that vanillin alone but had a synergistic effect on the E. coli O157:H7. Mixtures of cinnamon bark and vanillin had always a synergistic effect and some of the combination assayed showed bactericidal activity on the population of L. monocytogenes and E. coli O 157:H7.     

Growth and thermal inactivation of Listeria monocytogenes and Escherichia coli O157:H7 in four kinds of traditionally non-fermented soya bean products

This study evaluated growth and thermal inactivation of Listeria monocytogenes and Escherichia coli O157:H7 inoculated in tofu, dougan, qianzhang and doupi which were stored at 4, 25 and 37 °C and heated at 55, 60, 65 and 70 °C. Growth of the two pathogens in four soya bean products increased with temperature or A_w of soya bean products increasing. At the same temperature, lag time (LT) values of L. monocytogenes (16.32–0.94 h) and E. coli O157:H7 (2.66–0.98 h) in tofu which has the highest A_w were the lowest. When inoculated soya bean products were stored at 4 °C, L. monocytogenes grew slowly, while E. coli O157:H7 did not grow but survived for 14 days. With temperature increasing, δ-values of L. monocytogenes and E. coli O157:H7 in the four soya bean products were decreased. With A_w of soya bean products increasing, thermal resistance of L. monocytogenes decreased, while that of E. coli O157:H7 increased. This study could assist retail soya bean products processors and food industry to enhance safety of soya bean products and design thermal processing regimes.     

Analysis and the potential applications of essential oil and leaf extracts of <Emphasis Type="Italic">Silene armeria</Emphasis> L. to control food spoilage and food-borne pathogens

The aim of this study was to analyze the chemical composition of essential oil isolated from the floral parts of Silene armeria L. by hydrodistillation and to test the efficacy of essential oil and the various leaf extracts against a diverse range of microorganisms comprising food spoilage and food-borne pathogenic bacteria. The chemical composition of essential oil was analyzed by the GC–MS. It was determined that 28 compounds, which represented 89.03% of total oil, were present in the oil. The oil contained mainly methylamine (21.48%), β-butene (17.97%), α-butene (46.40%), coumaran (0.22%), eugenol (0.21%), α-humulene (0.07%), farnesol (0.05%) and linalool (0.12%). The essential oil (5 μl/ml, corresponding to 1,000 ppm/disc) and various leaf extracts of methanol, ethyl acetate, chloroform and hexane (7.5 μl/ml, corresponding to 1,500 ppm/disc) exhibited promising antibacterial effect against Bacillus subtilis ATCC6633, Listeria monocytogenes ATCC19166, Staphylococcus aureus KCTC1916, S. aureus ATCC6538, Pseudomonas aeruginosa KCTC2004, Salmonella typhimurium KCTC2515, Salmonella enteritidis KCTC2021, Escherichia coli O157-Human, E. coli ATCC8739, E. coli O57:H7 ATCC43888 and Enterobacter aerogenes KCTC2190. The zones of inhibition of different concentrations of essential oil and the various leaf extracts against the tested bacterial pathogens were found in the range of 10–19 and 7–13 mm, respectively, along with their respective MIC values ranging from 125 to 1,000 and 250–2,000 μg/ml. Also, the essential oil had a potential effect on the viable count of the tested bacteria. The results of this study suggest that the essential oil and leaf extracts derived from S. armeria could be used for the development of novel types of antibacterial agents to control food spoilage and food-borne pathogens.     

Determination of the effect of plant essential oils obtained by supercritical fluid extraction on the growth and viability of Listeria monocytogenes in broth and food systems using flow cytometry

The antimicrobial properties of oregano, rosemary and laurel extracts obtained by supercritical fluid extraction were investigated by examining their influence on the growth and viability of Listeria monocytogenes in laboratory medium and broccoli juice at 30 and 8 °C. Important decreases in the L. monocytogenes population were shown in presence of all the extracts obtained from rosemary and one oregano extract. The counts were reduced below the level of detection after 4 h of exposure at 30 °C in laboratory medium. A bactericidal effect was observed also when L. monocytogenes was exposed to rosemary at 30 °C and 8 °C in broccoli juice. Significant reductions in growth rate and an increase in lag phase of L. monocytogenes were observed in presence of some of the laurel and oregano extracts at both temperatures.Flow cytometry was used as a rapid method to determine the antibacterial effect of supercritical extracts and the physiological state of L. monocytogenes. Bacterial viability performed by dual staining of L. monocytogenes with SYTO 9 and propidium iodide revealed three different cell populations, specifically, living, dead and compromised cells. Live cell percentage decreased with the time of exposure, whereas the percentage of compromised cells remained constant and the dead cells increased in the same period.     

Antimicrobial activity of Thymus zygis essential oil against Listeria monocytogenes and its application as food preservative

Thymus zygis is an aromatic plant used in folk medicine. This work aimed to evaluate the anti-Listeria monocytogenes activity of T. zygis essential oil (EO), whose thymol is its major compound. Furthermore, the attenuation of this bacterium's virulence, namely by the inhibition of biofilm formation, motility and invasion of human cells, and the possible application of the EO in food were evaluated. The T. zygis EO showed antibacterial activity against L. monocytogenes with a minimum inhibitory concentration (MIC) of 0.05%, while showing a bactericidal effect. The EO significantly reduced the biofilm formation (inhibition from 16.85 to 89.86%) and motility (halos between 6.66 and 10.98 mm, compared to controls 13.12 to 17.22 mm), and not inducing cross-resistance to antibiotics, such as ampicillin, cefotaxime, erythromycin, gentamicin, tetracycline, and vancomycin. L. monocytogenes counts (initial inoculum of ~10⁶ CFU/mL) were lowered by the use of 2× MIC of T. zygis EO in the chicken juice (1.53 log CFU/mL) and lettuce model (to below the detection limit) after two days of storage. The use of EO (0.2% (v/v)) for sanitizing fresh vegetables, reduce L. monocytogenes and natural microbiota for values below the detection limit of the method for iceberg lettuce after an immersion of 5 min. For the spinach, L. monocytogenes was reduced in 4.35 log CFU/mL and the natural microbiota was diminished in a range of 4.47 to 5.94 log CFU/mL, when compared with the washing with water. Overall, the T. zygis EO has demonstrated a promising antimicrobial activity and these findings point to the potential of the EO as a natural food preservative or sanitizer for controlling L. monocytogenes in food products.     

Antimicrobial activity of recombinant human lactoferrin from <Emphasis Type="Italic">Aspergillus awamori</Emphasis>, human milk lactoferrin and their hydrolysates

The antibacterial activity of human lactoferrin from milk (hLF), recombinant human lactoferrin from Aspergillus awamori (rhLF) and their hydrolysates obtained with pepsin was investigated against Escherichia coli O157:H7, Salmonella Enteritidis and Listeria monocytogenes. The minimum inhibitory concentrations (MIC) and the minimum bactericidal concentrations (MBC) were determined for all the bacteria and the proteins assayed. Taking into account the MICs found for both lactoferrins studied, we can say that they behave very similarly, except for L. monocytogenes for which rhLF was more active. We studied the effect that heat treatments exerted on the antibacterial activity of the two types of lactoferrin and the only heat treatment that had a negative effect on that activity was 85 °C for 10 min. The activity of hLF and rhLF in UHT milk and whey against E. coli O157:H7 and L. monocytogenes, was also assayed. Our results showed a reduction in the number of viable cells for both microorganisms when were incubated with rhLF or hLF, but this decrease was lower than in broth media.     

Interpretation of the results of antimicrobial susceptibility analysis of Escherichia coli isolates from bovine milk,meat and associated foodstuffs

Strains of indicator bacteria Escherichia coli (E. coli) contaminating raw milk can, under specific conditions, become vectors of genes encoding resistance to antimicrobial drugs. The susceptibility of 1162 E. coli isolates originating from milk, milk products, meat, meat products and swabs from meat processing plant facilities to selected antimicrobial drugs (amikacin—AMI, ampicillin—AMP, cephalothin—CLT, cotrimoxazole—SXT, cefotaxime—CTX, gentamicin—GEN, neomycin—NEO, streptomycin—STR, tetracycline—TET, norfloxacin—NOR, chloramphenicol—CMP and erythromycin—ERY) was analysed. A standard microdilution method was used to detect minimal inhibition concentrations (MIC). The current criteria for interpretation (clinical breakpoints) do not allow identification of bacterial strains with acquired and potentially transmissible resistance to antimicrobial drugs. Microbiological breakpoints may be more appropriate because they take into account MIC distribution in a bacterial population of a certain species only. The aim of this study was to determine the microbiological breakpoints. Critical MIC_R values (microbiological breakpoints) were derived, which enabled E. coli strains to be characterized by MIC values into two subgroups; the first one comprises susceptible strains (MIC<MIC_R) and the second one comprises strains which are resistant to a given antimicrobial drug (MIC⩾MIC_R). We obtained the following values of microbiological breakpoints: AMI-32, AMP-32, CLT-64, CTX-8, CMP-32, GEN-8, ERY-⩾512, NEO-16, NOR-1, STR-32, SXT-0.8/15.2 and TET-16 μg/ml.     

Microbial profiles of frozen trimmings and silver sides prepared at Indian buffalo meat packing plants

To assess microbiological quality of buffalo meat trimmings (TT = 114) and silver sides (SS = 41), samples were collected from four different Indian meat packing plants. The aim of this study was: (i) to evaluate standard plate count (SPC), psychrotrophic count (PTC), Enterococcus feacalis count (EFC), Staphylococcus aureus count (SAC) and Escherichia coli count (ECC) and the presence of Salmonella spp. and Listeria monocytogenes; and (ii) also to determine vero toxic E. coli (VTEC) by polymerase chain reaction (PCR). TT samples had significantly higher (P < 0.001) SPC, PTC, EFC, and SAC than SS, while across the meat types there was no difference (P > 0.05) in ECC. E. coli was recovered from 32.4% TT and 19.5% SS samples. The prevalence rate of Salmonella spp. and L. monocytogenes in TT was 1.75% and 0.87%, respectively. But no SS sample was found to be positive for any of these two pathogens. VTEC was found in 2.58% of all the tested samples. This finding suggests that TT contain higher microbes but only small numbers of pathogens of latent zoonotic importance. The present study confirmed the importance of maintaining good process hygiene at meat plants for microbiological status of buffalo meat.     

The effect of American cranberry (Vaccinium macrocarpon) constituents on the growth inhibition,membrane integrity,and injury of Escherichia coli O157:H7 and Listeria monocytogenes in comparison to Lactobacillus rhamnosus

The antimicrobial properties of the American cranberry were studied against Escherichia coli O157:H7, Listeria monocytogenes, and Lactobacillus rhamnosus to determine the effects on growth inhibition, membrane permeability, and injury. Cranberry powder was separated using a C-18 Sep-Pak cartridge into sugars plus organic acids (F1), monomeric phenolics (F2), and anthocyanins plus proanthocyanidins (F3). Fraction 3 was further separated into anthocyanins (F4) and proanthocyanidins (F5) using an LH-20 Sephadex column. Each fraction was diluted in the brain heart infusion (BHI) broth to determine the minimum inhibitory/bactericidal concentrations (MIC/MBC). L. monocytogenes was the most susceptible to cranberry fraction treatment with the lowest MIC/MBC for each treatment, followed by E. coli O157:H7 and L. rhamnosus. Membrane permeability and potential was studied using LIVE/DEAD viability assay and using Bis (1, 3-dibutylbarbituric acid) trimethine oxonol (DiBAC₄), respectively. L. rhamnosus demonstrated the highest permeability followed by E. coli O157:H7, and L. monocytogenes. L. rhamnosus demonstrated the highest recovery followed by E. coli O157:H7, and L. monocytogenes. Each cranberry fraction demonstrated membrane hyperpolarization at their native pH, while F2, F3, and F5 demonstrated membrane depolarization at neutral pH. With this knowledge cranberry compounds may be used to prevent maladies and potentially substitute for synthetic preservatives and antibiotics.     

Quantifying Listeria monocytogenes prevalence and concentration in minced pork meat and estimating performance of three culture media from presence/absence microbiological testing using a deterministic and stochastic approach

Listeria monocytogenes poses a serious threat to public health, and the majority of cases of human listeriosis are associated with contaminated food. Reliable microbiological testing is needed for effective pathogen control by food industry and competent authorities. The aims of this work were to estimate the prevalence and concentration of L. monocytogenes in minced pork meat by the application of a Bayesian modeling approach, and also to determine the performance of three culture media commonly used for detecting L. monocytogenes in foods from a deterministic and stochastic perspective. Samples (n = 100) collected from local markets were tested for L. monocytogenes using in parallel the PALCAM, ALOA and RAPID'L.mono selective media according to ISO 11290-1:1996 and 11290-2:1998 methods. Presence of the pathogen was confirmed by conducting biochemical and molecular tests. Independent experiments (n = 10) for model validation purposes were performed. Performance attributes were calculated from the presence–absence microbiological test results by combining the results obtained from the culture media and confirmative tests. Dirichlet distribution, the multivariate expression of a Beta distribution, was used to analyze the performance data from a stochastic perspective. No L. monocytogenes was enumerated by direct-plating (<10 CFU/g), though the pathogen was detected in 22% of the samples. L. monocytogenes concentration was estimated at 14–17 CFU/kg. Validation showed good agreement between observed and predicted prevalence (error = −2.17%). The results showed that all media were best at ruling in L. monocytogenes presence than ruling it out. Sensitivity and specificity varied depending on the culture-dependent method. None of the culture media was perfect in detecting L. monocytogenes in minced pork meat alone. The use of at least two culture media in parallel enhanced the efficiency of L. monocytogenes detection. Bayesian modeling may reduce the time needed to draw conclusions regarding L. monocytogenes presence and the uncertainty of the results obtained. Furthermore, the problem of observing zero counts may be overcome by applying Bayesian analysis, making the determination of a test performance feasible.     

Effect of partial replacement of potassium lactate and sodium diacetate by natural green tea and grape seed extracts and postpackaging thermal treatment on the growth of Listeria monocytogenes in hotdog model system

Low‐ (5%) and high‐fat (20%) chicken and turkey hotdogs were formulated in three groups: no antimicrobials (control), chemical preservatives (potassium lactate and sodium diacetate) alone and partial replacement of chemical preservatives by green tea (GTE) and grape seed extracts (GSE), surface inoculated (c. 10³ CFU g^?1) with Listeria monocytogenes, treated with or without heat treatment (65 °C for 104 s) to determine the growth of L. monocytogenes until spoilage (28 days). Maximum growth inhibitions (c. 2.0 CFU g^?1) were observed in the treatments having chemical preservatives and plant extracts regardless of the meat and fat type. Furthermore, plant extracts along with chemical preservatives demonstrated additional inhibitory effect on the growth of L. monocytogenes survivors in chicken hotdog samples followed by postpackaging heat treatment. Results demonstrated that natural GTE and GSE can partially replace the chemical preservatives and further enhance the antilisterial activities when combined with heat treatment.     

Inhibition of biofilm formation,adhesion and invasion in Caco-2 cells by foodborne pathogens using phyto-mediated synthesised silver nanoparticles from industrial wastes

Certain foodborne diseases are associated with antibiotic resistance, a significant problem throughout the world. Silver nanoparticles (AgNPs) using industrial waste from Eucalyptus camaldulensis and sericin, a protein derived from Bombyx mori, were synthesised by a one-step approach. Spherical-shaped nanoparticles with the average size of 17.19 nm exhibited strong antioxidant activity. The minimum bactericidal concentrations against foodborne pathogens including Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7, Klebsiella pneumoniae, Salmonella Typhimurium, Shigella sonnei, Vibrio cholerae and Vibrio parahaemolyticus were between 2.96 and 11.83 µg/mL. Killing against L. monocytogenes and E. coli O157:H7 was observed within 4 h. Treatment with AgNPs at 0.25 – 0.5 × MIC significantly reduced biofilm production in all isolates (P < 0.05). AgNPs significantly impeded adhesion to and invasion of human epithelial Caco-2 cells by L. monocytogenes and E. coli O157:H7 (P < 0.05). Biocompatibility assessment of AgNPs with Caco-2 and human red blood cells demonstrated no toxic effects.     

Inhibition of Listeria monocytogenes by food antimicrobials applied singly and in combination

Abstract: Combining food antimicrobials can enhance inhibition of Listeria monocytogenes in ready-to-eat (RTE) meats. A broth dilution assay was used to compare the inhibition of L. monocytogenes resulting from exposure to nisin, acidic calcium sulfate, ɛ-poly-L-lysine, and lauric arginate ester applied singly and in combination. Minimum inhibitory concentrations (MICs) were the lowest concentrations of single antimicrobials producing inhibition following 24 h incubation at 35 °C. Minimum bactericidal concentrations (MBCs) were the lowest concentrations that decreased populations by ≥3.0 log₁₀ CFU/mL. Combinations of nisin with acidic calcium sulfate, nisin with lauric arginate ester, and ɛ-poly-L-lysine with acidic calcium sulfate were prepared using a checkerboard assay to determine optimal inhibitory combinations (OICs). Fractional inhibitory concentrations (FICs) were calculated from OICs and were used to create FIC indices (FIC_Is) and isobolograms to classify combinations as synergistic (FIC_I < 1.00), additive/indifferent (FIC_I= 1.00), or antagonistic (FIC_I > 1.00). MIC values for nisin ranged from 3.13 to 6.25 μg/g with MBC values at 6.25 μg/g for all strains except for Natl. Animal Disease Center (NADC) 2045. MIC values for ɛ-poly-L-lysine ranged from 6.25 to 12.50 μg/g with MBCs from 12.50 to 25.00 μg/g. Lauric arginate ester at 12.50 μg/g was the MIC and MBC for all strains; 12.50 mL/L was the MIC and MBC for acidic calcium sulfate. Combining nisin with acidic calcium sulfate synergistically inhibited L. monocytogenes; nisin with lauric arginate ester produced additive-type inhibition, while ɛ-poly-L-lysine with acidic calcium sulfate produced antagonistic-type inhibition. Applying nisin along with acidic calcium sulfate should be further investigated for efficacy on RTE meat surfaces. Practical Application: This study demonstrates the potential for combinations of antimicrobials to result in greater pathogen inhibition as compared to the application of a single antimicrobial. The data presented in this study can aid the food industry in developing more efficient and effective application of antimicrobials. These findings should also prompt further studies validating the inhibitory effect of combinations of antimicrobials on ready-to-eat surfaces.     

Nonaplex real-time PCR detection of Listeria monocytogenes, Campylobacter, Salmonella and enteropathogene E. coli after universal enrichment in food samples

A universal cultivation media for Listeria monocytogenes, Salmonella and enteropathogenic Escherichia coli in conjunction with a highly efficient nonaplex PCR for the determination of L. monocytogenes, Campylobacter, Salmonella and enteropathogenic E. coli was developed and compared to classical microbiological assays. The achieved detection limit was at 2 colony-forming units per g for all analytes. The method allows screening of food samples within 24 h and is partly automatable. A ring trial showed that it can be easily established in other laboratories showing its robustness. The presented method yields results similar to the classical ISO methods based on cultivation.     

Incorporation of Essential Oils and Nanoparticles in Pullulan Films to Control Foodborne Pathogens on Meat and Poultry Products

The incorporation of essential oils and nanotechnology into edible films has the potential to improve the microbiological safety of foods. The aim of this study was to evaluate the effectiveness of pullulan films containing essential oils and nanoparticles against 4 foodborne pathogens. Initial experiments using plate overlay assays demonstrated that 2% oregano essential oil was active against Staphylococcus aureus and Salmonella Typhimurium, whereas Listeria monocytogenes and Escherichia coli O157:H7 were not inhibited. Two percent rosemary essential oil was active against S. aureus, L. monocytogenes, E. coli O157:H7, and S. Typhimurium, when compared with 1%. Zinc oxide nanoparticles at 110 nm were active against S. aureus, L. monocytogenes, E. coli O157:H7, and S. Typhimurium, when compared with 100 or 130 nm. Conversely, 100 nm silver (Ag) nanoparticles were more active against S. aureus than L. monocytogenes. Using the results from these experiments, the compounds exhibiting the greatest activity were incorporated into pullulan films and found to inhibit all or some of the 4 pathogens in plate overlay assays. In challenge studies, pullulan films containing the compounds effectively inhibited the pathogens associated with vacuum packaged meat and poultry products stored at 4 °C for up to 3 wk, as compared to control films. Additionally, the structure and cross‐section of the films were evaluated using electron microscopy. The results from this study demonstrate that edible films made from pullulan and incorporated with essential oils or nanoparticles may improve the safety of refrigerated, fresh or further processed meat and poultry products.     

Antibacterial and antifungal activities of selected microalgae and cyanobacteria

In vitro activity of nine cyanobacterial and ten microalgal newly isolated or culture collection strains against eight significant food‐borne pathogens has been evaluated and compared. Water extracts and culture liquids of Gloeocapsa sp. and Synechocystis sp. demonstrated the widest spectrum of activity with minimal inhibitory concentration (MIC) ranging from 1.56 to 12.5 mg mL^?1. Culture liquid of Anabaena sp. had the highest activity (MIC = 0.39 mg mL^?1) but only to Gram‐positive bacteria. Ethanol extracts and fatty acids from all cyanobacteria and microalgae were active against Streptococcus pyogenes and/or Staphylococcus aureus. The fatty acids of Synechocystis sp. inhibited the growth of Bacillus cereus, Escherichia coli and Candida albicans (MIC values of 2.5–1.25 mg mL^?1, respectively). Exopolysaccharides (EPS) of Gloeocapsa sp. were the sample that exhibited activity against all test pathogens with lowest MIC values (0.125–1 mg mL^?1). High activity with a narrower range of susceptible targets demonstrated the exopolysaccharides of Synechocystis sp. and Rhodella reticulata. Antimicrobial activity was proven for phycobiliproteins isolated from Synechocystis sp., Arthrospira fusiformis, Porphyridium aerugineum and Porphyridium cruentum, respectively. In conclusion Gloeocapsa sp. and Synechocystis sp. and especially their exopolysaccharides showed the most promising potential against the examined food pathogens.     

Development of an Internal Amplification Control Using Multiplex PCR for the Detection of Listeria monocytogenes in Food Products

The bacterial pathogen Listeria monocytogenes is responsible for listeriosis, a food-borne disease, which may result in severe illness and possible death. Large outbreaks of listeriosis have been associated with food products including soft cheeses and ready to eat food products. Polymerase chain reaction (PCR) is a molecular identification method for food-borne pathogens; however, a drawback of this method is that false-positive or false-negative results may occur. To validate the accuracy of the PCR as a powerful molecular tool for pathogen detection, it is important that false-negative results be distinguishable from true-negative PCR results. The aim of this study was to design and include an internal amplification control (IAC) within the PCR to coamplify with L. monocytogenes in order to identify false-negative results of L. monocytogenes from ostrich meat and camembert cheese samples. The IAC had to be incorporated into the PCR without loss of specificity and sensitivity on the detection limit of L. monocytogenes and was developed and tested for use in a multiplex PCR detection system. A region of the pUC19 plasmid was selected as the IAC for this study. The optimal concentration at which pUC19 would coamplify with L. monocytogenes was determined to be 0.001 pg/μL. Following an enrichment procedure, the minimum number of organisms detected in a spiked food sample by the PCR was 8 CFU/mL L. monocytogenes; the same detection limit was attained when the pUC19 IAC was included in the PCR. An optimal pUC19 IAC concentration increased the reliability of the PCR for food diagnostic purposes.     

Reductionin Listeria monocytogenes,Salmonella enterica and Escherichia coli O157:H7 in vitro and on tomato by sophorolipid and sanitiser as affected by temperature and storage time

Increased consumption of produce by consumers has been attributed to perceived health benefits of postharvest produce. Pathogen control is crucial because periodic occurrences and contamination of tomato and leafy greens have exacerbated food safety risks for consumers. We investigated the effects of temperatures (5 and 25 °C), storage time (30 min and 24 h) for inactivation of Listeria monocytogenes, Salmonella enterica and Escherichia coli O157:H7 by sophorolipid (SL‐p) produced fermentatively using palmitic acid as a co‐substrate at different concentrations in vitro. Reduction in pathogenic bacteria on grape tomato by SL‐p, sanitiser (Lovit) and combinations of SL‐p and sanitiser was determined. Temperature and storage time significantly (P < 0.05) affected pathogen inactivations by SL‐p as pathogen reductions were greater at 25 °C and 24 h than at 5 °C and 30 min of storage. L. monocytogenes was the most sensitive to SL‐p treatment as reductions of 5 log relative to untreated controls were attained at 0.12% of SL‐p. Significant reductions in S. enterica (1.91–3.85 logs) and E. coli O157:H7 (0.87–4.09 logs) were recorded at 2–5% of SL‐p. Lower populations of Salmonella and E. coli O157:H7 were inactivated than L. monocytogenes. On grape tomato, pathogen populations inactivated increased at higher SL‐p levels at 25 °C. Sanitiser and sanitiser + SL‐p reduced bacterial populations on tomato by 5.29–5.76 logs and 0.71–3.3.66 logs, respectively. These results imply the interactions of temperature, storage time and SL‐p significantly (P < 0.05) affected pathogen strain reductions. The combination of SL‐p with sanitiser led to synergistic effect on E. coli O157:H7, but not L. monocytogenes and S. enterica.     

Synergistic antimicrobial activity of galangal (Alpinia galanga), rosemary (Rosmarinus officinalis) and lemon iron bark (Eucalyptus staigerana) extracts

BACKGROUND: In this study the synergistic antimicrobial activities of combinations of extracts from galangal (Alpinia galanga), rosemary (Rosmarinus officinalis) and lemon iron bark (Eucalyptus staigerana) were evaluated against Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, Salmonella typhimurium and Clostridium perfringens. Chemical compositions of these extracts were also determined to provide further insight into antimicrobial constituents and their potential mechanisms of action. RESULTS: Combinations of galangal with either rosemary or lemon iron bark showed synergistic antimicrobial activity. Specifically, galangal and rosemary showed synergistic activity against S. aureus and L. monocytogenes only, while galangal and lemon iron bark showed synergistic activity against E. coli and S. typhimurium. Chemical compositions of the extracts were determined by gas chromatographic–mass spectrometric analysis. The major chemical components of the galangal and lemon iron bark extracts were 1′‐acetoxy‐chavicol acetate (1′ACA) (63.4%) and neral (15.6%), respectively, while 1,8‐cineole (26.3%) and camphor (20.3%) were identified as major chemical components of the rosemary extract. CONCLUSION: The results of this study show that galangal, rosemary and lemon iron bark extracts contain components that may have different modes of antimicrobial action and combinations of these extracts may have potential as natural antimicrobials to preserve foods. Copyright © 2010 Society of Chemical Industry