The effect of micro-architectural structure of cabbage substratum and or background bacterial flora on the growth of Listeria monocytogenes

The effect of micro-architectural structure of cabbage (Brassica oleracea var. capitata L.) substratum and or background bacterial flora on the growth of Listeria monocytogenes as a function of incubation temperature was investigated. A cocktail mixture of Pseudomonas fluorescens, Pantoea agglomerans and Lactobacillus plantarum was constituted to a population density of approximately 5 log CFU/ml in order to pseudo-simulate background bacterial flora of fresh-cut cabbage. This mixture was co-inoculated with L. monocytogenes (approximately 3 log CFU/ml) on fresh-cut cabbage or in autoclaved cabbage juice followed by incubation at different temperatures (4-30 degrees C). Data on growth of L. monocytogenes were fitted to the primary growth model of Baranyi in order to generate the growth kinetic parameters of the pathogen. During storage, microbial ecology was dominated by P. fluorescens and L. plantarum at refrigeration and abuse temperature, respectively. At all temperatures investigated, lag duration (lambda, h), maximum specific growth rate (micro(max), h(-1)) and maximum population density (MPD, log CFU/ml) of L. monocytogenes were only affected by medium micro-architectural structure, except at 4 degrees C where it had no effect on the micro(max) of the pathogen. Comparison of observed values of micro(max) with those obtained from the Pathogen Modelling Program (PMP), showed that PMP overestimated the growth rate of L. monocytogenes on fresh-cut cabbage and in cabbage juice, respectively. Temperature dependency of micro(max) of L. monocytogenes, according to the models of Ratkowsky and Arrhenius, showed linearity for temperature range of 4-15 degrees C, discontinuities and linearity again for temperature range of 20-30 degrees C. The results of this experiment have shown that the constituted background bacterial flora had no effect on the growth of L. monocytogenes and that micro-architectural structure of the vegetable was the primary factor that limited the applicability of PMP model for predicting the growth of L. monocytogenes on fresh-cut cabbage. A major limitation of this study however is that nutrient profile of the autoclaved cabbage juice may be different from that of the raw juice thus compromising realistic comparison of the behaviour of L. monocytogenes as affected by micro-architectural structure.     

Rapid detection of Salmonella in pet food: design and evaluation of integrated methods based on real-time PCR detection

Reducing the risk of Salmonella contamination in pet food is critical for both companion animals and humans, and its importance is reflected by the substantial increase in the demand for pathogen testing. Accurate and rapid detection of foodborne pathogens improves food safety, protects the public health, and benefits food producers by assuring product quality while facilitating product release in a timely manner. Traditional culture-based methods for Salmonella screening are laborious and can take 5 to 7 days to obtain definitive results. In this study, we developed two methods for the detection of low levels of Salmonella in pet food using real-time PCR: (i) detection of Salmonella in 25 g of dried pet food in less than 14 h with an automated magnetic bead-based nucleic acid extraction method and (ii) detection of Salmonella in 375 g of composite dry pet food matrix in less than 24 h with a manual centrifugation-based nucleic acid preparation method. Both methods included a preclarification step using a novel protocol that removes food matrix-associated debris and PCR inhibitors and improves the sensitivity of detection. Validation studies revealed no significant differences between the two real-time PCR methods and the standard U.S. Food and Drug Administration Bacteriological Analytical Manual (chapter 5) culture confirmation method.     

Nonstarter lactobacilli isolated from soft and semihard Argentinean cheeses: genetic characterization and resistance to biological barriers

Nonstarter lactic acid bacteria isolated from Argentinean cheeses were identified and characterized by focusing on their resistance to biological barriers, along with other physiological features of potential interest, in the search for future probiotic organisms. Lactobacilli were enumerated and isolated from semihard and soft cheeses made with multistrain Streptococcus thermophilus starters. Lactobacilli counts in 1-week-old cheeses were between 10(5) and 10(7) CFU/g and then reached 10(7) CFU/ g in all 1-month samples, while streptococci were always above 10(9) CFU/g. A total number of 22 lactobacilli isolates were retained, identified, and characterized by in vitro tests. Species identity was determined by carbohydrate metabolism and species-specific PCR assays. Genetic diversity was explored by random amplified polymorphic DNA (RAPD) PCR analysis. The Lactobacillus strains were assigned to the species L. casei, L. plantarum, L. rhamnosus, L. curvatus, L. fermentum, and L. perolens. All the strains studied tolerated 25 ppm of lysozyme, and most of them showed resistance to 0.3% bile. After incubation in gastric solution (pH 2.0), counts decreased by several log units, ranging from 3.2 to 7.0. The strains were able to grow in the presence of bile salts, but only three isolates were capable of deconjugation. The nonstarter lactobacilli that were assayed fermented the prebiotic substrates (especially lactulose and inulin). Some strains showed high cell hydrophobicity and beta-galactosidase activity, as well as inhibitory activity against pathogenic bacteria. It was concluded that most of the lactobacilli isolated in this study demonstrated resistance to biological barriers and physiological characteristics compatible with probiotic properties, which make them suitable for further research in in vivo studies aimed at identifying new probiotic organisms.     

Chitosan effects on quality properties of Greek style fresh pork sausages

The effect of chitosan (0.5% and 1%) added individually or in combination with nitrites (150 ppm) on microbiological (Total Viable Counts, Lactic acid bacteria, Pseudomonas spp., Brochothrix thermosphacta, Enterobacteriaceae, yeasts and moulds), physicochemical-chemical (pH, chemical composition, lipid oxidation) and sensory properties of fresh pork sausages stored at 4 °C for 28 days was investigated. Chitosan addition resulted in significant (p < 0.05) inhibition of microbial growth, while nitrites did not seem to protect sausages from microbial spoilage. A gradual reduction of nitrites was observed till the end of storage, when nitrites were almost depleted in all nitrite containing samples. The rate of lipid oxidation in fresh pork sausages was significantly decreased (p < 0.05) by addition of increasing levels of chitosan, while samples containing both chitosan and nitrites showed the lowest malondialdehyde (MDA) values, indicating a synergistic antioxidative effect. Consequently, the samples containing the combination of nitrites and chitosan at any level deteriorated less rapidly and were judged as more acceptable than all the other samples.     

Development and application of Multiple-Locus Variable number of tandem repeat Analysis (MLVA) to subtype a collection of Listeria monocytogenes

In this study we report the development and application of a Multiple-Locus Variable number of tandem repeat Analysis (MLVA) strategy for subtyping Listeria monocytogenes. Genome profiles of a collection of forty-five food-borne L. monocytogenes isolates were compared using MLVA. These isolates were obtained as part of an active surveillance programme of foods in the south-east region of Ireland. MLVA successfully discriminated amongst the isolates. The method was easy to perform, relatively fast and could be deployed in any molecular laboratory with basic laboratory equipment. This approach is a valuable tool, which has the capability to provide comparable results when compared with other more established typing methods, including pulsed-field gel electrophoresis (PFGE).     

Isolation and pulsed-field gel electrophoresis typing of Listeria monocytogenes from modified atmosphere packaged fresh-cut vegetables collected in Ireland

The incidence of Listeria monocytogenes in modified atmosphere packaged fresh-cut fruits and vegetables from chill cabinets of a supermarket in Ireland was investigated over a 2-year period. Overall, 9.58% of fresh-cut produce was contaminated with Listeria spp. Various species of Listeria were isolated from samples, including L. monocytogenes, L. seeligeri, L. innocua, L. welshimeri, and L. ivanovii. No fruit samples contained detectable L. monocytogenes. Overall, a total of 21 L. monocytogenes isolates (2.9% of samples) were recovered from a range of products, including dry coleslaw mix (80% shredded cabbage and 20% shredded carrot), bean sprouts, and leafy vegetables such iceberg, romaine, and radicchio lettuce and mixed salad leaves (curly endive, escarole, and radicchio leaves). Dry coleslaw mix appeared to have the highest incidence of Listeria contamination (20%) compared with other products. Listeria contamination was more frequent (P < 0.05) during the summer and autumn months than during the winter and spring months. The 21 L. monocytogenes isolates were subsequently subtyped by genomic macrorestriction techniques using ApaI with pulsed-field gel electrophoresis (PFGE). PFGE of digested DNA produced bands of 79 to 518 kb. Four PFGE profiles were identified, and approximately 50% of the isolates were associated with profile 1. This study indicates that fresh-cut vegetables packaged under a modified atmosphere can support growth of numerous species of Listeria, including L. monocytogenes.     

Uncertainty analysis of the use of a retailer fidelity card scheme in the assessment of food additive intake

The feasibility of using a retailer fidelity card scheme to estimate food additive intake was investigated in an earlier study. Fidelity card survey information was combined with information provided by the retailer on levels of the food colour Sunset Yellow (E110) in the foods to estimate a daily exposure to the additive in the Swiss population. As with any dietary exposure method the fidelity card scheme is subject to uncertainties and in this paper the impact of uncertainties associated with input variables including the amounts of food purchased, the levels of E110 in food, the proportion of food purchased at the retailer, the rate of fidelity card usage, the proportion of foods consumed outside of the home and bodyweights and with systematic uncertainties was assessed using a qualitative, deterministic and probabilistic approach. An analysis of the sensitivity of the results to each of the probabilistic inputs was also undertaken. The analysis identified the key factors responsible for uncertainty within the model and demonstrated how the application of some simple probabilistic approaches can be used quantitatively to assess uncertainty.     

Effect of a nonionic surfactant on biodegradation of slowly desorbing PAHs in contaminated soils

The influence of the nonionic surfactant Brij 35 on biodegradation of slowly desorbing polycyclic aromatic hydrocarbons (PAHs) was determined in contaminated soils. We employed a soil originated from a creosote-polluted site, and a manufactured gas plant soil that had been treated by bioremediation. The two soils differed in their total content in five indicator 3-, 4-, and 5-ring PAHs (2923 mg kg(-1) and 183 mg kg(-1) in the creosote-polluted and bioremediated soils, respectively) but had a similar content (140 mg kg(-1) vs 156 mg kg(-1)) of slowly desorbing PAHs. The PAHs present in the bioremediated soil were highly recalcitrant. The surfactant at a concentration above its critical micelle concentration enhanced the biodegradation of slowly desorbing PAHs in suspensions of both soils, but it was especially efficient with bioremediated soil, causing a 62% loss of the total PAH content. An inhibition of biodegradation was observed with the high-molecular-weight PAHs pyrene and benzo[a]pyrene in the untreated soil, possibly due to competition effects with other solubilized PAHs present at relatively high concentrations. We suggest that nonionic surfactants may improve bioremediation performance with soils that have previously undergone extensive bioremediation to enrich for a slowly desorbing profile.     

Levels and distribution of polybrominated diphenyl ethers in water, surface sediments, and bivalves from the San Francisco Estuary

Polybrominated diphenyl ethers (PBDEs) were found in water, surface sediments, and bivalve samples that were collected from the San Francisco Estuary in 2002. sigmaPBDE concentrations in water samples ranged from 3 to 513 pg/ L, with the highest concentrations found in the Lower South Bay (range 103-513 pg/L) region, which receives approximately 26% of the Estuary's wastewater treatment plant effluents. The sigmaPBDEs in sediments ranged from below detection limits to 212 ng/g dry wt, with the highest concentration found at a South Bay station (212 ng/g dry wt), which was up to 3 orders of magnitude higher than other stations. The sigmaPBDE concentrations ranged from 9 to 64 ng/g dry wt in oysters (Crassostrea gigas), from 13 to 47 ng/g dry wt in mussels (Mytilus californianus), and from 85 to 106 ng/g dry wt in clams (Corbicula fluminea). Only three PBDE congeners were detected in bivalves, BDE-47, BDE-99, and BOE-100; these are the most bioaccumulative congeners from the commercial Penta-BDE mixture.