Efficacy of plant essential oils against foodborne pathogens and spoilage bacteria associated with ready-to-eat vegetables: antimicrobial and sensory screening

The objectives of this study were to evaluate the antimicrobial activity of plant essential oils (EOs) against foodborne pathogens and key spoilage bacteria pertinent to ready-to-eat vegetables and to screen the selected EOs for sensory acceptability. The EOs basil, caraway, fennel, lemon balm, marjoram, nutmeg, oregano, parsley, rosemary, sage, and thyme were evaluated. The bacteria evaluated were Listeria spp., Staphylococcus aureus, Lactobacillus spp., Bacillus cereus, Salmonella, Enterobacter spp., Escherichia coli, and Pseudomonas spp. Quantitative antimicrobial analyses were performed using an absorbance-based microplate assay. Efficacy was compared using MIC, the half maximum inhibitory concentration, and the increase in lag phase. Generally, gram-positive bacteria were more sensitive to EOs than were gram-negative bacteria, and Listeria monocytogenes strains were among the most sensitive. Of the spoilage organisms, Pseudomonas spp. were the most resistant. Oregano and thyme EOs had the highest activity against all the tested bacteria. Marjoram and basil EOs had selectively high activity against B. cereus, Enterobacter aerogenes, E. coli, and Salmonella, and lemon balm and sage EOs had adequate activity against L. monocytogenes and S. aureus. Within bacterial species, EO efficacy was dependent on strain and in some cases the origin of the strain. On a carrot model product, basil, lemon balm, marjoram, oregano, and thyme EOs were deemed organoleptically acceptable, but only oregano and marjoram EOs were deemed acceptable for lettuce. Selected EOs may be useful as natural and safe additives for promoting the safety and quality of ready-to-eat vegetables.     

Antimicrobial activity of plant essential oils using food model media: Efficacy,synergistic potential and interactions with food components

The aim of this study was to optimise the antimicrobial efficacy of plant essential oils (EOs) for control of Listeria spp. and spoilage bacteria using food model media based on lettuce, meat and milk. The EOs evaluated were lemon balm, marjoram, oregano and thyme and their minimum inhibitory concentrations (MIC) were determined against Enterobacter spp., Listeria spp., Lactobacillus spp., and Pseudomonas spp. using the agar dilution method and/or the absorbance based microplate assay. MICs were significantly lower in lettuce and beef media than in TSB. Listeria strains were more sensitive than spoilage bacteria, and oregano and thyme were the most active EOs. EO combinations were investigated using the checkerboard method and Oregano combined with thyme had additive effects against spoilage organisms. Combining lemon balm with thyme yielded additive activity against Listeria strains. The effect of simple sugars and pH on antimicrobial efficacy of oregano and thyme was assessed in a beef extract and tomato serum model media. EOs retained greater efficacy at pH 5 and 2.32% sugar, but sugar concentrations above 5% did not negatively impact EO efficacy. In addition to proven antimicrobial efficacy, careful selection and investigation of EOs appropriate to the sensory profile of foods and composition of the food system is required. This work shows that EOs might be more effective against food-borne pathogens and spoilage bacteria when applied to foods containing a high protein level at acidic pH, as well as moderate levels of simple sugars.     

Antibacterial activity of extracts from some edible plants commonly consumed in Asia

Extracts of edible plants (26 species) from China, Japan, Thailand and Yemen were screened for their antibacterial activity against Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli and Salmonella infantis. Buffered methanol (80% methanol and 20% PBS) and acetone extracted inhibitory substances against tested bacteria from 16 plants, as revealed by the disc assay. The minimum inhibitory concentrations (MICs) of extracts determined by the agar dilution method ranged from 165 to 2640 mg l⁻¹. The most sensitive microorganism to extracts from Azadirachta indica, Cinnamomum cassia, Rumex nervosus, Ruta graveolens, Thymus serpyllum and Zingiber officinale was B. cereus, with MIC of 165 to 660 mg l⁻¹. E. coli and S. infantis were only inhibited by Cinnamomum cassia extracts at the highest MIC (2640 mg l⁻¹). L. monocytogenes (Tottori) was more resistant than the ATCC 7644 strain to extracts from Ruta chalepensis, Artemisia absinthium and Cissus spp. EDTA (0.85 mM) reduced the MICs of Cinnamomum cassia and Cissus rotundifolia by at least 50% when tested against E. coli, S. infantis, S. aureus and L. monocytogenes.     

Antioxidant and Antimicrobial Properties of Selected Spice Extracts

Tinctures of eleven plants used as spices (basil, celery, dill, horsetail, lovage, marjoram, milfoil, oregano, parsley, rosemary, and thyme) were tested for their antiradical properties by means of the DPPH (1,1-diphenyl-2-picrylhydrazyl) assay over a two year period. Seven of these plants (basil, lovage, marjoram, milfoil, oregano, rosemary and thyme) were selected to obtain a mixture, which was tested in situ as antioxidant on vegetable and animal fats by an accelerated oxidation test at 110°C. The herbal extract also showed antimicrobial activity against Staphylococcus aureus (ATCC 25923), Streptococcus pyogenes (ATCC 49399), Escherichia coli (ATCC 25922) and Candida albicans (ATCC 24433).     

Antimicrobial activity of whey protein isolate edible films with essential oils against food spoilers and foodborne pathogens

The use of antimicrobial edible film is proposed as a means of improving food safety and extending the shelf-life of food systems by controlling the release of antimicrobials on food surfaces. In this work we first selected and studied 8 different essential oils (EOs) from plants, namely, oregano, clove, tea tree, coriander, mastic thyme, laurel, rosemary, and sage as natural antimicrobials against 2 gram-positive bacteria (Listeria innocua and Staphylococcus aureus) and 2 gram-negative bacteria (Salmonella enteritidis and Pseudomona fragi) by using the agar disk diffusion method. EOs from oregano, clove, and tea tree produced the largest surfaces of inhibition against the growth of the 4 bacterial strains tested. Second and following the assessment of compatibility, stable antimicrobial edible films based on whey protein isolate (WPI) with increasing concentrations (0.5% to 9%) of the 8 EOs were developed and tested for antimicrobial activity against the same gram-positive and gram-negative bacteria. WPI-edible films incorporating oregano or clove EO were found to have the most intense inhibitory effect of microbial growth. The bacterial strain gram-negative P. fragi presented the less susceptibility to the effect of those films. Moreover, only the edible films based on these 2 EOs were active against all 4 studied microorganisms. On the other hand, the edible films incorporating tea tree, coriander, mastic thyme, laurel, rosemary, or sage EOs even at high concentrations (7% to 9%) did not cause any antimicrobial effect against the pathogens S. aureus or S. enteritidis. PRACTICAL APPLICATION: Potential applications of this technology can introduce direct benefits to the food industry by improving safety and microbial product quality. The results of this research have direct application in the food industry with potential applications in various foodstuffs, including meat and poultry products where the control of spoilage bacteria such as P. fragi throughout their chilled storage or the improvement of food safety by controlling pathogens such as S. enteritidis are topics of particular interest for the industry.     

Synergistic antimicrobial activity of two binary combinations of marjoram,lavender, and wild thyme essential oils

The combination of essential oils (EOs) is a novel alternative to improve their preservative effects and to reduce their organoleptic impact in food. In this context, this work aims to investigate the antibacterial combined effect of two EOs combinations through the calculation of the fractional inhibitory concentration index. The combinations tested consists of Lavandula dentata/Origanum majorana EOs and Thymus serpyllum/Origanum majorana EOs. Their chemical compositions were identified by CG/MS analyses. The main compounds of O. majorana EO were (-) - terpinene-4-ol and trans-4-thujanol. Those of L. dentata EO were β-pinene and 1,8-cineole, and those of T. serpyllum EO were p-cymene and γ-terpinene. Regarding the outcomes, results highlighted partial synergistic and additive interactions. Two combinations of marjoram and thyme EOs had antibacterial activities against S. aureus. The first one corresponded to the quarter of the minimum inhibitory concentration of marjoram and half that of thyme. The second one was the mix of half and quarter of the minimum inhibitory concentration of respectively marjoram and thyme EOs. This last combination also showed an antibacterial effect against E. coli. The quarter and the half of their minimum inhibitory concentration of marjoram and lavender combination, respectively, gave a partial synergy against both strains. Henceforth, these findings could be largely exploited in food preservation through the use of minimal doses of these plant products without affecting the antibacterial and the organoleptic properties in foods.     

Antimicrobial effect of spices and herbs on Shigella sonnei and Shigella flexneri

Of 17 spices and herbs tested at 1% (wt/vol) in Mueller-Hinton (MH) agar, only cloves, thyme, oregano, allspice, basil, rosemary, and marjoram showed antimicrobial effects on Shigella. The MICs of thyme, oregano, basil, and rosemary (as determined by the agar dilution method) ranged from 0.5 to 1% (wt/vol) depending on the Shigella strain used. With the use of various combinations of temperatures (12, 22, and 37 degrees C), pHs (5.0, 5.5, and 6.0), and NaCl concentrations (1, 2, 3, and 4%, wt/vol) and the inclusion or exclusion of thyme or basil at 1% (wt/vol) in an MH agar model system, it was established that basil or thyme can contribute to combination processing as a growth-inhibitory factor for Shigella spp. In the presence of basil and thyme, Shigella flexneri did not develop CFU during the 7-day incubation period for, respectively, 14 and 16 of the 18 tested combinations, while growth was noted in the corresponding temperature-pH-NaCl concentration combinations without basil or thyme. A growth-inhibitory effect on Shigella sonnei was also noted. The results of an orientation study involving the addition of basil and thyme to spaghetti sauce prior to autoclaving and S. sonnei inoculation indicated that basil and thyme contributed to the reduction of S. sonnei after 16 days at 12 degrees C but not at 4 degrees C.     

Interactive inhibition of meat spoilage and pathogenic bacteria by lysozyme,nisin and EDTA in the presence of nitrite and sodium chloride at 24 degrees C

To develop a nisin- and lysozyme-based antimicrobial treatment for use with processed ham and bologna, in vitro experiments were conducted to determine whether inhibition enhancing interactions occur between the antimicrobials lysozyme, chrisin (a commercial nisin preparation), EDTA, NaCl and NaNO₂. Inhibitory interactions were observed between a number of agents when used against specific pathogenic and spoilage bacteria. The observed interactions included lysozyme with EDTA (Enterococcus faecalis and Weissella viridescens), chrisin with EDTA (all Gram-positive organisms), EDTA with NaCl (Escherichia coli, Salmonella enterica serovar Typhimurium, Serratia grimesii), EDTA with nitrite (E. coli, Lactobacillus curvatus, Leuconostoc mesenteroides, Listeria monocytogenes, S. Typhimurium), chrisin with nitrite (Lc. mesenteroides, L. monocytogenes), and NaCl with nitrite (S. Typhimurium, Shewanella putrefaciens). Previous reports have described interactions between nisin with EDTA that resulted in enhanced antimicrobial effect against Gram-negative bacteria, or lysozyme with nisin against Gram-positive bacteria. These interactions were not observed in these experiments. We observed that unlike previous studies, these experiments were conducted on growing cells in nutrient broth, rather than under conditions of nutrient limitation. We propose that screening of antimicrobials for use in food systems in nutrient-deficient systems is inappropriate and that new protocols should be developed.     

Survival of pathogenic microorganisms in an egg-nog-like product containing 7% ethanol

A liquor consisting of whole egg, saccharose (25% w/v) and ethanol (7.0% w/v) was artificially contaminated with Salmonella enteritidis, S. typhimurium, Staphylococcus aureus (three different strains), Bacillus cereus and Listeria monocytogenes. After 3 weeks of incubation at 22°C the numbers of Salmonella, S. aureus and L. monocytogenes decreased more than 3 log₁₀ units. Under such conditions, however, the total number of microorganisms increased 3 log₁₀ units. At 4°C the decrease of pathogenic microorganisms was much slower and a decrease of 3 log₁₀ units was observed only after 7 weeks of incubation. Egg-nog, without ethanol, incubated at 22°C allowed growth of Salmonella and S. aureus, while the numbers of B. cereus spores remained unchanged. Vegetative cells of B. cereus as well as L. monocytogenes decreased in numbers. However, after prolonged incubation the numbers of L. monocytogenes increased significantly.     

Assessment of the influence of some spice extracts on the formation of heterocyclic amines in meat

Beef meat samples were heated in the presence of basil, oregano, marjoram, rosemary, sweet grass, savory, thyme and coriander extracts and the effects on the formation of heterocyclic amines (HAs) were measured by HPLC with fluorescence detection. The results obtained after heating meat in diethylene glycol showed that the addition of 0.2% and 0.5% of thyme, savory and oregano extracts slightly decreased the formation of 1-methyl-6-phenyl-1H-imidazo[4,5-b]pyridin-2-amine (PhIP). Other extracts did not have any influence on the concentration of PhIP, while in some cases they promoted its formation, particularly with basil extract. Oregano and basil extracts, which showed the best positive and negative effects on the formation of PhIP, were tested by heating meat in vegetable oil. Both extracts increased PhIP amount, but decreased Trp-P-2 concentration (see Section 2.1). The content of Trp-P-1 decreased in the samples with basil and increased in those with oregano. No correlation between the anti-oxidant activity of extracts and the formation of PhIP was found.     

Inhibition of Listeria monocytogenes by Elite Clonal Extracts of Oregano (Origanum vulgare)

Food safety continues to be a major concern for the food industry in recent years. One of the industry's top priorities has been to find alternative ways to preserve their newly developed foods while satisfying the increasing consumer demand to produce safe, all-natural products. In order to achieve this “clean label”, much research has been devoted to the use of effective plant-based antimicrobials, such as those from herbs and spices, to replace chemical preservatives. However, due to the cross-pollination character of herbs and spices, there is a lot of genetic heterogeneity among different batches of the same plant species. This poses a problem for the routine use of plants, and their extracts, as a barrier towards microbial growth. In order to combat this, a unique tissue-culture-based selection strategy was used to isolate an elite phenolic phytochemical-producing clonal line of oregano (Origanum vulgare). Ethanol extracts of this elite clonal line of oregano were then used to study its inhibitory action against Listeria monocytogenes in both broth and meat systems. Thymol and carvacrol, two of the main phenolic constituents of oregano extracts, were also tested in both systems to evaluate their activity against that of the whole oregano extract.

Results indicate that thymol, carvacrol, and the clonal oregano line were all effective in inhibiting the growth of L. monocytogenes in both systems. Approximately 150-200 ppm of pure carvacrol or thymol was needed in order to significantly inhibit the growth of L. monocytogenes in broth, while at least 1200 ppm (corresponding to 27.8 μg phenolics/ml) of the elite clonal oregano extract was needed to do the same. Inconclusive results were obtained when the clonal line was compared to store-brand samples of oregano. In meat systems, 800 ppm of the oregano extract was able to significantly inhibit the growth of the pathogen more so than 800 ppm of carvacrol. A possible explanation for this is that the oregano extract was able to work more effectively at the interface of the lipid and water-soluble portions of the meat than the carvacrol. These results are promising for the food industry since we have now developed an approach for a highly consistent and reliable natural source of antimicrobial activity for future studies.     

Impact of Essential Oils on the Taste Acceptance of Tomato Juice,Vegetable Soup,or Poultry Burgers

Despite the vast body of available literature on the possibilities of essential oils (EOs) as food preservatives or functional ingredients, the sensory impact of their addition to foods has barely been approached. This work focuses on the hedonic taste acceptance of 3 food products (tomato juice, vegetable soup, and poultry burgers) when they are incorporated with potentially antimicrobial concentrations (20 to 200 μL/L) of 6 selected EOs (lemon, pennyroyal mint, thyme, and rosemary) and individual compounds (carvacrol, p‐cymene). Although addition of 20 μL/L of pennyroyal mint or lemon EO did not change the taste acceptance of tomato juice, higher concentrations of these compounds or any concentration of the other 4 compounds did. In vegetable soup, the tolerance limit for rosemary EO, thyme EO, carvacrol, or p‐cymene was 20 μL/L, while the addition of 200 μL/L of lemon EO was accepted. Tolerance limits in poultry burgers were established in 20 μL/L for carvacrol and thyme EOs, 100 μL/L for pennyroyal mint EO and p‐cymene, and 200 μL/L for lemon and rosemary EOs. Moreover, incorporation of pennyroyal mint EO to tomato juice or poultry burgers, and enrichment of vegetable soup with lemon EO, could contribute to the development of food products with an improved sensory appeal.     

Characterization of the Effect of Sprouting or Solid-State Bioprocessing by Dietary Fungus on the Antibacterial Activity of Soybean Extracts Against Listeria monocytogenes

Listeria monocytogenes is one of the most severe food-borne bacterial infections causing Listeriosis. As L. monocytogenes can survive harsh adverse conditions - such as low pH, high NaCl, and refrigeration temperatures - as well as resist current antimicrobial measures such as the use of disinfectants and antibiotics, there is a need for alternative anti-Listeria strategies. In the search for new antimicrobial agents, much recent research has focused on the potential of dietary phenolic compounds. In this study, soybean extracts enriched for phenolic content via dark-germination sprouting or solid-state bioprocessing by the dietary fungus Rhizopus oligosporus or Lentinus edodes were investigated for in vitro antibacterial activity against L. monocytogenes.L. monocytogenes growth was inhibited most effectively by R. oligosporus bioprocessed soybean extracts, which showed anti-Listeria activity at total phenolic concentrations as low as 10 µg 100 µL^-1. In both sprouted soybean extract and L. edodes-bioprocessed soybean extract the anti-Listeria activity was not observed until at least 200 µg total phenolic content 100 µL^-1 was used. Anti-Listeria activity by soybean extract was associated with phenolic mobilization but not with antioxidant activity. Further, R. oligosporus bioprocessed soybean extracts were shown to inhibit the growth of L. monocytogenes in fish and meat systems at refrigeration temperatures. The potential involvement of mobilization of antimicrobial versus non-antimicrobial phenolics during sprouting and solid-state bioprocessing was hypothesized and discussed.     

Effect of drying method on the antioxidant capacity of six Lamiaceae herbs

The present study investigated the changes in total phenols (TP), rosmarinic acid content and antioxidant capacity of six Lamiaceae herbs (rosemary, oregano, marjoram, sage, basil and thyme) after three drying treatments (air-, freeze- and vacuum oven-drying) stored for 60 days at −20 °C and compared to fresh samples. Ferric reducing antioxidant property (FRAP) and oxygen radical absorbance capacity (ORAC) were used as markers for antioxidant capacity. Air-dried samples had significantly (p < 0.05) higher TP, rosmarinic acid content and antioxidant capacity than had freeze-dried and vacuum oven-dried samples throughout the storage period. Fresh samples had the lowest values for the parameters tested. Vacuum oven-drying resulted in higher TP and FRAP values in rosemary and thyme during 60 days of storage than did freeze-drying. In ORAC assay, the difference was significantly higher only in thyme. Storage did not show any effect on the dried samples for the parameters tested.     

Comparing predicting models for heat inactivation of Listeria monocytogenes and Pseudomonas aeruginosa at different pH

Under the same experimental conditions it has been demonstrated that whereas survival curves of Listeria monocytogenes in the range of temperatures from 54 to 62 °C followed a first-order kinetic, those of Pseudomonas aeruginosa in the range of temperatures from 50 to 56 °C were not linear showing a shoulder followed by a linear region. The first order kinetic model did not describe survival curves of P. aeruginosa. A model based on the Weibull distribution (Log₁₀(N_t/N₀)=(1/−2.303)*(t/b)ⁿ)) accurately described the inactivation kinetics of both microorganisms at the three pHs of 4, 5.5, 7.4 investigated. For both microorganisms, the b value depended on the treatment temperature and the pH of the treatment medium. Whereas for L. monocytogenes the n value was independent of the treatment conditions, for P. aeruginosa the n value depended on the pH of the treatment medium.

The model based on the Weibull distribution was capable of accurately predicting the treatment time to inactivate five Log₁₀ cycles of both microorganisms at the three pHs investigated.     

Characterization of Listeria spp. isolated from ready-to-eat products in Argentina using SDS–PAGE and restriction endonuclease

Listeria spp. are considered of interest in public health since their presence indicates the potencial existence of L. monocytogenes. Total cellular proteins and DNA from four strains of L. monocytogenes serotype 4, four strains of L. monocytogenes belonging to serotype 1, twelve strains of L. innocua, four strains of L. seeligeri and two strains of L. welshimeri isolated from ready–to–eat food were studied by SDS–PAGE and restriction endonuclease digestion. SDS–PAGE protein profiles obtained were species specific and could be evaluated by visual comparison. Enzyme for restriction endonuclease analysis was EcoRI, discriminating L. monocytogenes from other Listeria spp. These methodologies might be a helpful tool and a good alternative for epidemiological tracking of listeriosis in laboratories, where other methods are not available.     

Microbiological quality of 18 degrees C ready-to-eat food products sold in Taiwan

A total of 164 samples of 18 °C ready-to-eat (RTE) food products, purchased in 1999–2000 from convenience stores and supermarkets in central Taiwan, were examined to determine the microbiological quality of these products. The 18 °C RTE food products, manufactured by 16 factories, were divided into groups based on the type of food and their major ingredients. Aerobic plate count, coliforms, Escherichia coli, Bacillus cereus, Staphylococcus aureus and psychrotrophic Pseudomonas spp. were evaluated. The incidence of E. coli and coliforms in these 18 °C RTE food products was 7.9% and 75.0%, respectively, while 49.8% and 17.9% of the samples were found to contain B. cereus and S. aureus, respectively. Among the samples tested, 1.3% of the food products contained more than 10⁵ CFU g⁻¹ of B. cereus and 0.7% contained more than 10⁵ CFU g⁻¹ of S. aureus. The pH values of the samples were all below 7.0, except for cold noodles, which had pH values ranging from 5.18 to 8.20. Among the five types of 18 °C food products tested, the highest incidence of E. coli (16%) and Pseudomonas spp. (64.0%) were detected in hand-rolled sushi in a cone shape. On the other hand, the highest incidence rate of coliforms, B. cereus, and S. aureus were found in sandwiches (88%), cold noodles (66.7%) and rice balls rolled in seaweed (25.0%), respectively. Food products made of ham contained the highest incidence of coliforms (88.0%) and E. coli (16.0%), while food products containing meat and ham as the major ingredients had the highest incidence rates of B. cereus (62.5%) and S. aureus (26.1%), respectively. For coliforms, E. coli, B. cereus and S. aureus, the percentage of 18 °C RTE food products exceeding the microbiological standards for RTE food accepted by Republic of China was 75.0%, 7.9%, 49.8% and 17.9%, respectively.     

Detection and Rapid Purification of Internalin B as a Protein Marker in Listeria monocytogenes

Clinical and food strains of Listeria monocytogenes have been found to express InternalinB (InlB) without polymorphism. InlB, which is a 67-kDa surface protein, behaves as an invasion and adhesion protein of the bacterium into cells. Thus, InlB could be a good candidate as a protein marker to detect L. monocytogenes. Three strains of L. monocytogenes (ATCC 19115, serotype 4b, ATCC 19111, serotype 1/2a, ATCC 7644, serotype 1/2c) were tested to detect and purify InlB. L. grayii (ATCC 25400) and L. innocua (isolated from soft cheese) were used as controls that did not express InlB due to the absence of its gene on the chromosome. InlB was quantitatively extracted in a solubilized form by treatment of L. monocytogenes with 1 M Tris-Cl at pH 7.5. Immunoblot analysis using anti-InlB polyclonal antibody revealed that L. monocytogenes 19115 had the lowest expression, which required enrichment of InlB for its detection. Simple spin-ion exchange chromatography with strong acidic cation exchanger was used to enrich the InlB protein that is strongly basic. Most impurities in the column were washed with 25 mM sodium acetate whereas the InlB protein was only protein retained in the column and can be eluted by 1 M NaCl. The data presented here showed that spin-ion exchange chromatography was found to be a simple and rapid method to enrich and purify InlB within 20 min.     

Caffeic acid derivatives in dried Lamiaceae and Echinacea purpurea products

The concentrations of caffeic acid derivatives within Lamiaceae and Echinacea (herb, spice, tea, and dietary supplement forms) readily available in the US marketplace (n = 72) were determined. After the first identification of chicoric acid in Ocimum basilicum (basil), the extent to which chicoric acid could be found within the family Lamiaceae was investigated. The dominant phenolic acid in all Lamiaceae samples was rosmarinic acid, which ranged from 2.04 mg/100 g (one of 12 oregano samples) to 622.28 mg/100 g (lemon balm). Of the herbs tested in this study (marjoram, oregano, peppermint, rosemary, sage, spearmint, and thyme from the family Lamiaceae), only basil and lemon balm (Melissa officinalis) contained chicoric acid. Basil samples (starting material and resulting end product) obtained from an industry cooperator, showed substantial phenolic deficiency as a result of processing (approximately 78% loss).     

Comparison of the hydrophobic grid-membrane filter DNA probe method and the Health Protection Branch standard method for the detection of Listeria monocytogenes in foods

The standard Health Protection Branch (HPB) method for the detection of L. monocytogenes in foods involves lengthy enrichment, selection and biochemical testing, requiring up to 8 days to complete. A hydrophobic grid-membrane filter (HGMF) method employing a digoxigenin-labelled listeriolysin O probe required 5 days to complete, and included an image-analysis system for electronic data acquisition. A total of 200 food samples encompassing 8 high-risk food groups (soft and semi-soft cheeses, packaged raw vegetables, frozen cooked shrimp, ground poultry, ground pork, ground beef, jellied meats, and pâté) were screened for the presence of L. monocytogenes by the two methods. Overall, 32 (16%) and 30 (15%) of the naturally-contaminated food samples tested positive for L. monocytogenes by the HPB and DNA methods, respectively. The DNA probe method was highly specific in discriminating L. monocytogenes from other Listeria spp. present in 50 of the samples tested. Results showed 94% sensitivity and 100% specificity between the two methods. The HGMF DNA probe method is an efficient and reliable alternative to the HPB standard method for detecting L. monocytogenes in foods.