The effect of four alternative chilling regimes on the bacterial load on beef carcasses.

The objective of this study was to compare the effect of current (10 °C for 10 h followed by 0 °C with a low fan speed) versus four alternative beef carcass chilling regimes, ranging from -6 °C to 0 °C and wind speeds between 1.5 and 6 m/s on the microbiology of beef carcasses. The temperature and relative humidity (RH) in the chillers, the carcass core and surface temperature, pH, water activity (aw) and carcass weight (drip) loss were recorded. Bacterial concentrations (total viable counts (TVC), total Enterobacteriaceae counts (TEC), Pseudomonas spp., lactic acid bacteria (LAB) and Brochothrix thermosphacta) were also monitored. Similar pH, aw and drip loss (2%) values were obtained regardless of chilling regime. For the most part, bacterial concentrations were also similar and, where statistically significant (P < 0.05) counts occurred, the reductions were low (≤1 log10 cfu/cm2). It was concluded that the current chilling regime was as effective as the tested alternatives in terms of the bacterial quality of the carcasses.

Identification of biofilm hotspots in a meat processing environment: Detection of spoilage bacteria in multi-species biofilms.

Biofilms are comprised of microorganisms embedded in a self-produced matrix that normally adhere to a surface. In the food processing environment they are suggested to be a source of contamination leading to food spoilage or the transmission of food-borne pathogens. To date, research has mainly focused on the presence of (biofilm-forming) bacteria within food processing environments, without measuring the associated biofilm matrix components. Here, we assessed the presence of biofilms within a meat processing environment, processing pork, poultry and beef, by the detection of microorganisms and at least two biofilm matrix components. Sampling included 47 food contact surfaces and 61 non-food contact surfaces from eleven rooms within an Austrian meat processing plant, either during operation or after cleaning and disinfection. The 108 samples were analysed for the presence of microorganisms by cultivation and targeted quantitative real-time PCR based on 16S rRNA. Furthermore, the presence of the major matrix components carbohydrates, extracellular DNA and proteins was evaluated. Overall, we identified ten biofilm hotspots, among them seven of which were sampled during operation and three after cleaning and disinfection. Five biofilms were detected on food contact surfaces (cutters and associated equipment and a screw conveyor) and five on non-food contact surfaces (drains and water hoses) resulting in 9.3 % of the sites being classified as biofilm positive. From these biofilm positive samples, we cultivated bacteria of 29 different genera. The most prevalent bacteria belonged to the genera Brochothrix (present in 80 % of biofilms), Pseudomonas and Psychrobacter (isolated from 70 % biofilms). From each biofilm we isolated bacteria from four to twelve different genera, indicating the presence of multi-species biofilms. This work ultimately determined the presence of multi-species biofilms within the meat processing environment, thereby identifying various sources of potential contamination. Especially the identification of biofilms in water hoses and associated parts highlights the need of a frequent monitoring at these sites. The knowledge gained about the presence and composition of biofilms (i.e. chemical and microbiological) will help to prevent and reduce biofilm formation within food processing environments.

qPCR quantification of Carnobacterium maltaromaticum, Brochothrix thermosphacta, and Serratia liquefaciens growth kinetics in mixed culture.

Quantifying growth kinetics of specific spoilage microorganisms in mixed culture is required to describe the evolution of food microbiomes. A qPCR method was developed to selectively amplify individual meat spoilage bacteria, Carnobacterium maltaromaticum, Brochothrix thermosphacta and Serratia liquefaciens, within a broth medium designed to simulate the composition of beef. An optimized method of DNA extraction was produced for standard curve construction. Method specificity was determined by individual single peaks in melt curves. Reaction efficiency for standard curves of C. maltaromaticum, B. thermosphacta and S. liquefaciens was high (R2 = 0.98-0.99), and linear quantification was achieved over a 5 log CFU/ml range. Coefficient of variation was calculated considering both threshold cycle (Ct) and bacterial concentration; the value did not exceed 14% for inter- or intra-runs for either method. Comparison of growth kinetic parameters derived from plate count and qPCR showed no significant variation (P > .05) for growth rate (GR) and maximum population density (MPD); lag phase duration (LPD) was not included in this comparison due to high innate variability. Log quantification of each isolate was validated in a mixed-culture experiment for all three species with qPCR and plate count differing less than 0.3 log CFU/ml (average 0.10 log CFU/ml, R2 = 0.98).

The complex multicellular morphology of the food spoilage bacteria Brochothrix thermosphacta strains isolated from ground chicken.

Herein we describe a highly structured, filamentous growth phenotype displayed by an isolate of the food spoilage microorganism Brochothrix thermosphacta. The growth morphology of this B. thermosphacta strain (strain BII) was dependent on environmental factors such as the growth media, incubation temperatures, and the inoculum concentration. Inoculation of cultures in highly dilute suspensions resulted in the formation of isolated, tight aggregates resembling fungal growth in liquid media. This same strain also formed stable, mesh-like structures in 6-well tissue culture plates under specific growth conditions. The complex growth phenotype does not appear to be unique to strain BII but was common among B. thermosphacta strains isolated from chicken. Light and electron micrographs showed that the filaments of multiple BII cells can organize into complex, tertiary structures resembling multistranded cables. Time-lapse microscopy was employed to monitor the development of such aggregates over 18 h and revealed growth originating from short filaments into compact ball-like clusters that appeared fuzzy due to protruding filaments or cables. This report is the first to document this complex filamentous growth phenotype in a wild-type bacterial isolate of B. thermosphacta.

Analysis of microbiome in raw chicken meat from butcher shops and packaged products in South Korea to detect the potential risk of foodborne illness.

Chicken meat is one of the most widely consumed meats worldwide. The microbiota on the whole body of chicken is a potential source of foodborne pathogens that can be transmitted to humans during the preparation of raw meat. However, to date, there have been no studies comparing the microbiota of packaged chicken products and those of raw chicken carcasses from butcher shops, although such information could be useful for identifying sources of contamination in cases of food poisoning. We addressed this in the present study by analyzing the microbiota of 80 chicken meat samples collected from various butcher shops and processing plants in South Korea with the Illumina MiSeq system based on the 16S rRNA gene sequence. The bacterial amounts in chicken samples were estimated by quantitative real-time PCR. Although different microbial members were present in unpackaged meat from butcher shops as compared to those in packaged products from commercial sources, seasonal differences (sample obtained in January vs. July) in microbiota were more significant even in the packaged products from the same company. We also investigated the influence of contaminated foodborne pathogen on the indigenous microbiota (64 chicken samples) by artificially inoculated with Salmonella enterica serotype Virchow on chicken carcasses under various conditions, and carrying out 16S rRNA gene and whole metagenome sequencing. The amount of contaminated Salmonella in chicken meat samples was the highest and lowest in samples stored at 27 °C and 4 °C after washing, respectively. Additionally, the relative abundance of virulence genes was detected lower in samples stored at 4 °C after washing in both butcher shop and commercial samples. These results could be useful for reducing the risk of foodborne illness caused by cross-contamination during the preparation of chicken meat.

Genotypic and phenotypic characterization of the food spoilage bacterium Brochothrix thermosphacta.

Microbial food spoilage is responsible for significant economic losses. Brochothrix thermosphacta is one of the major bacteria involved in the spoilage of meat and seafood. Its growth and metabolic activities during food storage result in the production of metabolites associated with off-odors. In this study, we evaluated the genotypic and phenotypic diversity of this species. A collection of 161 B. thermosphacta strains isolated from different foods, spoiled or not, and from a slaughterhouse environment was constituted from various laboratory collections and completed with new isolates. A PCR test based on the rpoB gene was developed for a fast screening of B. thermosphacta isolates. Strains were typed by MALDI-TOF MS, rep-PCR, and PFGE. Each typing method separated strains into distinct groups, revealing significant intra-species diversity. These classifications did not correlate with the ecological origin of strains. The ability to produce acetoin and diacetyl, two molecules associated with B. thermosphacta spoilage, was evaluated in meat and shrimp juices. The production level was variable between strains and the spoilage ability on meat or shrimp juice did not correlate with the substrate origin of strains. Although the B. thermosphacta species encompasses ubiquitous strains, spoiling ability is both strain- and environment-dependent.

Dynamics of bacterial communities and interaction networks in thawed fish fillets during chilled storage in air.

Thawed hake (Merluccius capensis and M. paradoxus) and plaice (Pleuronectes platessa) fillets were used as a model to evaluate the effect of storage temperature (0 or 10 °C) and biological variability (fish species, lot to lot) on bacterial growth kinetics and microbial successions. Both culture dependent methods (plate counts on non-selective and selective media) and culture independent methods (qPCR and 16S rRNA gene metabarcoding) were used. Bacterial counts exceeded 107 cfu/g within 2-3 days at 10 °C and 7-8 days at 0 °C. Plate counts on three media (Plate Count Agar +0.5% NaCl, Iron Agar Lyngby and Pseudomonas Selective medium) and 16S rRNA gene counts estimated by qPCR were highly correlated. Growth was modelled using the D-model and specific growth rate ranged between 0.97 and 1.24 d-1 and 3.54 and 5.90 d-1 at 0 and 10 °C, respectively. The initial composition of the microbiota showed lot-to-lot variation, but significant differences between the two fish species were detected. Alpha diversity significantly decreased during storage. When bacterial counts exceeded 107 cfu/g, the microbiota was dominated by members of the genera Pseudomonas, Psychrobacter, Acinetobacter, Serratia, Flavobacterium, Acinetobacter, Carnobacterium, Brochothrix and Vagococcus. However, Photobacterium and Shewanella, two genera frequently associated with fish spoilage, were either absent or minor components of the microbiota. As expected, storage temperature significantly affected the abundance of several species. The inference of microbial association networks with three different approaches (an ensemble approach using the CoNet app, Sparse Correlations for Compositional data, and SParse InversE Covariance Estimation for Ecological Association Inference) allowed the detection of both a core microbiota, which was present throughout storage, and a number of taxa, which became dominant at the end of spoilage and were characterized by a disproportionate amount of negative interactions.

Influence of lactate and acetate removal on the microbiota of French fresh pork sausages.

The microbiota of fresh French pork sausages were characterised in five batches of comminuted pork meat that were equally divided into two formulations either containing the acid-based preservatives lactate and acetate, or no preservatives. Conventional microbiological analysis and high-throughput 16S rDNA amplicon sequencing methods were performed on meat batches packed under modified atmosphere (70% oxygen and 30% carbon dioxide) during chilled storage. In addition, meat pH and colour, and gas composition of the packages were monitored until the end of the shelf-life. During storage, the population of mesophilic and lactic acid bacteria increased from 4 log CFU/g to 8 log CFU/g after 15 days of chilled storage, both with and without preservatives. Despite similar changes of the physical and chemical parameters, such as pH and package gas composition, spoilage was delayed in the meat containing the preservatives, suggesting that lactate and acetate are effective against spoilage. Metagenetic analysis showed that at the end of the shelf-life, the species distribution differed between both the formulations and the batches. Lactic acid bacteria were shown to dominate both with and without preservatives; however, samples containing no preservatives were characterised by the presence of an increased population of Brochothrix spp. and Pseudomonas spp. whereas, Leuconostoc mesenteroides/pseudomesenteroides and Lactobacillus curvatus/graminis were more abundant in the meat with preservatives.

Exploring lot-to-lot variation in spoilage bacterial communities on commercial modified atmosphere packaged beef.

Understanding the factors influencing meat bacterial communities is important as these communities are largely responsible for meat spoilage. The composition and structure of a bacterial community on a high-O2 modified-atmosphere packaged beef product were examined after packaging, on the use-by date and two days after, to determine whether the communities at each stage were similar to those in samples taken from different production lots. Furthermore, we examined whether the taxa associated with product spoilage were distributed across production lots. Results from 16S rRNA amplicon sequencing showed that while the early samples harbored distinct bacterial communities, after 8-12 days storage at 6 °C the communities were similar to those in samples from different lots, comprising mainly of common meat spoilage bacteria Carnobacterium spp., Brochothrix spp., Leuconostoc spp. and Lactococcus spp. Interestingly, abundant operational taxonomic units associated with product spoilage were shared between the production lots, suggesting that the bacteria enable to spoil the product were constant contaminants in the production chain. A characteristic succession pattern and the distribution of common spoilage bacteria between lots suggest that both the packaging type and the initial community structure influenced the development of the spoilage bacterial community.

Comparison of hot versus cold boning of beef carcasses on bacterial growth and the risk of blown pack spoilage.

Primals were prepared from beef Longissimus thoracis et lumborum (LTL), psoas major (PM), quadriceps femoris (QF) and semitendinosus (S) muscles from cold and hot boned carcasses, vacuum-packaged and stored for 42 or 100days at 2°C and 7°C. Storage temperature, carcass or primal surface temperature, pH and aw were monitored. Samples were taken periodically and tested for total viable count mesophilic (TVCm), TVC psychrophilic (TVCp), total Enterobacteriaceae count (TEC), presumptive Pseudomonas spp., lactic acid bacteria (LAB), Clostridium spp. and Brochothrix thermosphacta. A fifth muscle, biceps femoris (BF), was used to examine the impact of hot boning on blown pack spoilage (BPS). Primal counts increased to 6-7log10cfucm-2 after 6weeks. Significantly (P<0.05) higher TEC, Pseudomonas spp. and Br. thermosphacta counts were observed on cold versus hot boned primals. In contrast, significantly (P<0.05) higher TVC, LAB and Clostridium spp. concentrations were obtained on hot boned beef. Moreover, BPS pack distension/bursting occurred considerably sooner in hot boned product.

The microbiology of beef carcasses and primals during chilling and commercial storage.

The primary objective of this study was to characterise (microbiology and physical parameters) beef carcasses and primals during chilled storage. A minor aim was to compare observed growth of key spoilage bacteria on carcasses with that predicted by ComBase and the Food Safety Spoilage Predictor (FSSP). Total viable count (TVC), total Enterobacteriacae count (TEC), Pseudomonas spp., lactic acid bacteria (LAB), Brochothrix thermosphacta and Clostridium spp. were monitored on beef carcasses (n = 30) and primals (n = 105) during chilled storage using EC Decision 2001/471/EC and ISO sampling/laboratory procedures. The surface and/or core temperature, pH and water activity (aw) were also recorded. Clostridium (1.89 log10 cfu/cm2) and Pseudomonas spp. (2.12 log10 cfu/cm2) were initially the most prevalent bacteria on carcasses and primals, respectively. The shortest mean generation time (G) was observed on carcasses with Br. thermosphacta (20.3 h) and on primals with LAB (G = 68.8 h) and Clostridium spp. (G = 67 h). Over the course of the experiment the surface temperature decreased from 37 °C to 0 °C, pH from 7.07 to 5.65 and aw from 0.97 to 0.93 The observed Pseudomonas spp. and Br. thermosphacta growth was more or less within the range of predictions of Combase. In contrast, the FSSP completely overestimated the growth of LAB. This study contributes to the very limited microbiological data on beef carcasses and primals during chilling.

Quality assessment of ice-stored tropical yellowfin tuna (Thunnus albacares) and influence of vacuum and modified atmosphere packaging.

Metagenomic, microbial, chemical and sensory analyses of Thunnus albacares from Martinique stored in ice (AIR - 0 °C), vacuum (VP - 4/8 °C) and modified atmosphere packaging (MAP - 4/8 °C) (70% CO2 - 30% O2) were carried out. The organoleptic rejection of AIR tuna was observed at day 13 when total bacterial counts equaled 10(6)-10(7) CFU g(-1). No extension of shelf-life was provided by VP and MAP. According to 16S rRNA gene sequence analyzed by Illumina MiSeq and PCR-TTGE, Rhodanobacter terrae was the main species of the freshly caught tuna. At the sensory rejection time, Brochothrix thermosphacta and Pseudomonas dominated the AIR products while B. thermosphacta alone or a mix of B. thermosphacta, Enterobacteriaceae and lactic acid bacteria (LAB) dominated the microbiota of MAP and VP products, respectively. The pH value remained stable in all trials, ranging from 5.77 to 5.97. Total volatile basic nitrogen (TVBN) and trimethylamine (TMA-N) concentrations were weak and not significantly different between batches. Lipid oxidation increased in the samples containing O2 (MAP > AIR). The initial concentration of histamine was high (75-78 mg kg(-1)) and stable up to 8 days but then significantly decreased in all trials to reach 25-30 mg kg(-1), probably due to the presence of histamine-decomposing bacteria.

Identification and growth dynamics of meat spoilage microorganisms in modified atmosphere packaged poultry meat by MALDI-TOF MS.

Modified atmosphere packaging (MAP) is widely used in food industry to extend the microbiological shelf-life of meat. Typically, poultry meat has been packaged in a CO2/N2 atmosphere (with residual low O2). Recently, some producers use high O2 MAP for poultry meat to empirically reach comparable shelf lifes. In this work, we compared spoilage microbiota of skinless chicken breast in high (80% O2, 20% CO2) and low O2 MAP (65% N2 and 35% CO2). Two batches of meat were incubated in each atmosphere for 14 days at 4 °C and 10 °C. Atmospheric composition of each pack and colony forming units (25 °C, 48 h, BHI agar) of poultry samples were determined at seven timepoints. Identification of spoilage organisms was carried out by MALDI-TOF MS. Brochothrix thermosphacta, Carnobacterium sp. and Pseudomonas sp. were the main organisms found after eight days at 4 °C and 10 °C in high O2 MAP. In low O2 MAP, the main spoilage microbiota was represented by species Hafnia alvei at 10 °C, and genera Carnobacterium sp., Serratia sp., and Yersinia sp. at 4 °C. High O2 MAP is suggested as preferential gas because were less detrimental and pathogens like Yersinia were not observed.

Spoilage potential of Vagococcus salmoninarum in preservative-free, MAP-stored brown shrimp and differentiation from Brochothrix thermosphacta on streptomycin thallous acetate actidione agar.

AIMS: During a previous study concerning brown shrimp (Crangon crangon), selective streptomycin thallous acetate actidione (STAA) agar was used to determine the growth of Brochothrix thermosphacta. However, the growth of Vagococcus salmoninarum on this medium was also noticed. This study explores the spoilage potential of this organism when inoculated on sterile shrimp. METHODS AND RESULTS: Isolates growing on STAA were identified using (GTG)5 clustering followed by partial 16S rRNA gene sequence analysis. Their biochemical spoilage potential was analysed for H2 S production and enzymatic activities were tested using an APIZYM test. Headspace solid phase micro-extraction (SPME) and gas chromatography-mass spectrometry (GC-MS) were used to analyse the volatile organic compounds (VOCs) produced during storage of inoculated shrimp. CONCLUSION: Fifty-five per cent of isolates taken from STAA could be identified as V. salmoninarum, while no apparent morphological difference with B. thermosphacta isolates was identified upon the prescribed incubation conditions. For isolates identified as V. salmoninarum, production of 2-heptanone, 2-nonanone, 2-undecanone was found, as was the possibility to form H2 S. SIGNIFICANCE AND IMPACT OF THE STUDY: When using the STAA medium for detecting B. thermosphacta, one should consider the possible abundant presence of V. salmoninarum as well. Based on this study, V. salmoninarum does not exhibit great spoilage potential, although it can produce H2 S and formed VOCs which are also found in other spoiled seafood products.

Microbiological spoilage and volatiles production of gutted European sea bass stored under air and commercial modified atmosphere package at 2 °C.

Microbiological, sensory, TVB-N and TMA-N changes and Volatile Organic Compounds (VOCs) detection using the SPME/GC-MS technique, were performed to evaluate potential chemical spoilage indices (CSI) of gutted sea bass (Dicentrarchus labrax) stored at 2 °C under air and in modified atmosphere packaging (MAP CO2: 60%, O2: 10%, N2: 30%). Shelf-life, determined by sensory evaluation, of gutted sea bass stored at 2 °C under air and MAP was 9 and 13 d respectively. Pseudomonas and H2S producing bacteria were among the dominant spoilage microorganisms under both storage conditions, while Lactic Acid Bacteria (LAB) and Brochothrix thermosphacta were co-dominant with Pseudomonas and H2S producing bacteria under MAP. The traditional CSIs such as TVB-N and TMA-N were increased substantially only at the late stages of storage or after rejection of the products, making them unsuitable for freshness/spoilage monitoring throughout storage. A substantial number of VOCs attributed to microbiological action or chemical activity, were detected including alcohols, aldehydes, ketones, organic acids and esters. The level of microbial origin VOCs such as ethanol, 2-ethyl-1-hexanol, 3-methyl-1-butanol, 2-methyl-1-butanol, 3-methylbutanal, 2-methylbutanal and some ethyl esters increased during storage, suggesting their potential as CSIs.

The spoilage characteristics of Brochothrix thermosphacta and two psychrotolerant Enterobacteriacae in vacuum packed lamb and the comparison between high and low pH cuts.

The spoilage potential of Brochothrix thermosphacta, Serratia proteamaculans and Rahnella aquatilis was investigated in vacuum packaged high (5.9 to 6.4) and low (5.4 to 5.8) pH lamb. Vacuum packaged fore shank (m. extensor carpi radialis) and striploins (m. longissimus dorsi) (n=306) inoculated with ~100 CFU of individual bacteria were stored for twelve weeks at temperatures -1.5, 0, 2 and 7°C. Spoilage characteristics and bacterial numbers were recorded and analysed in comparison to un-inoculated control samples. All three bacterial species were shown to grow in vacuum packaged lamb of pH values between 5.4 and 6.4, when stored at chilled temperatures (-1.5 to 7°C) for up to 84 days. B. thermosphacta and S. proteamaculans caused spoilage to the meat under these conditions whilst R. aquatilis spoiled high pH meat at 7°C. These results go against previous beef models stipulating that Brochothrix and Enterobacteriacae species cannot grow on or cause spoilage of low pH meat in the absence of oxygen.

Nondestructive measurement of total volatile basic nitrogen (TVB-N) in pork meat by integrating near infrared spectroscopy, computer vision and electronic nose techniques.

Total volatile basic nitrogen (TVB-N) content is an important reference index for evaluating pork freshness. This paper attempted to measure TVB-N content in pork meat using integrating near infrared spectroscopy (NIRS), computer vision (CV), and electronic nose (E-nose) techniques. In the experiment, 90 pork samples with different freshness were collected for data acquisition by three different techniques, respectively. Then, the individual characteristic variables were extracted from each sensor. Next, principal component analysis (PCA) was used to achieve data fusion based on these characteristic variables from 3 different sensors data. Back-propagation artificial neural network (BP-ANN) was used to construct the model for TVB-N content prediction, and the top principal components (PCs) were extracted as the input of model. The result of the model was achieved as follows: the root mean square error of prediction (RMSEP) = 2.73 mg/100g and the determination coefficient (R(p)(2)) = 0.9527 in the prediction set. Compared with single technique, integrating three techniques, in this paper, has its own superiority. This work demonstrates that it has the potential in nondestructive detection of TVB-N content in pork meat using integrating NIRS, CV and E-nose, and data fusion from multi-technique could significantly improve TVB-N prediction performance.

The near-quantitative sampling of genomic DNA from various food-borne Eubacteria.

BACKGROUND: The disruption of the bacterial cell wall plays an important part in achieving quantitative extraction of DNA from Eubacteria essential for accurate analyses of genetic material recovered from environmental samples. RESULTS: In this work we have tested a dozen commercial bacterial genomic DNA extraction methodologies on an average of 7.70 × 10(6) (±9.05%), 4.77 × 10(8) (±31.0%), and 5.93 × 10(8) (±4.69%) colony forming units (CFU) associated with 3 cultures (n = 3) each of Brochothrix thermosphacta (Bt; Gram-positive), Shigella sonnei (Ss; Gram-negative), and Escherichia coli O79 (Ec; Gram-negative). We have utilized real-time PCR (qPCR) quantification with two specific sets of primers associated with the 16S rRNA "gene" to determine the number of copies CFU(-1) by comparing the unknown target DNA qPCR results with standards for each primer set. Based upon statistical analyses of our results, we determined that the Agencourt Genfind v2, High Pure PCR Template Prep Kit, and Omnilyse methods consistently provided the best yield of genomic DNA ranging from 141 to 934, 8 to 21, and 16 to 27 16S rDNA copies CFU(-1) for Bt, Ss, and Ec. If one assumes 6-7 copies of the 16S rRNA gene per genome, between 1 and 3 genomes per actively dividing cell and ≥ 100 cells CFU(-1) for Bt (found to be a reasonable assumption using an optical method expounded upon herein) or between 1 and 2 cells CFU(-1) for either Ss or Ec, then the Omnilyse procedure provided nearly quantitative extraction of genomic DNA from these isolates (934 ± 19.9 copies CFU(-1) for Bt; 20.8 ± 2.68 copies CFU(-1) for Ss; 26.9 ± 3.39 copies CFU(-1) for Ec). The Agencourt, High Pure, and Omnilyse technologies were subsequently assessed using 5 additional Gram-positive and 10 Gram-negative foodborne isolates (n = 3) using a set of "universal" 16S rDNA primers. CONCLUSION: Overall, the most notable DNA extraction method was found to be the Omnilyse procedure which is a "bead blender" technology involving high frequency agitation in the presence of zirconium silicate beads.

Spoilage characteristics of Brochothrix thermosphacta and campestris in chilled vacuum packaged lamb, and their detection and identification by real time PCR.

The spoilage potential of Brochothrix campestris and Brochothrix thermosphacta was investigated in vacuum-packed lamb. Striploins (n=338) were inoculated and stored for twelve weeks at temperatures -1.5, 0, 2 and 7 °C. Growth around 5-6 log10 CFU/cm(2) was recorded after six weeks at 0, 2 and 7 °C, and ~3 log10 CFU/cm(2) after nine weeks at -1.5 °C. B. campestris was shown to cause spoilage by nine weeks at temperatures above 0 °C by the presence of green drip and unacceptable odours. Molecular based assays for the detection and differentiation of B. thermosphacta and B. campestris were developed and validated. A TaqMan assay was designed to target a unique single-nucleotide polymorphism in the Brochothrix 16s rRNA gene with a sensitivity of <7 CFU per reaction. Secondly a specific PCR was designed for B. campestris targeting the structural genes, brcA and brcB. These testing regimes offer a rapid and cost effective method for the detection and screening of Brochothrix species in meat products and processing environments.

Effect of thyme oil on the preservation of vacuum-packaged chicken liver.

In this study fresh chicken liver meat was stored under vacuum packaging (VP) and under refrigeration (4 °C). The following treatments were used: V (control samples, stored under VP), VT1 (thyme oil; 0.1% v/w, stored under VP) and VT2 (thyme oil; 0.3% v/w, stored under VP). Lipid oxidation was low, as judged by determination of malondialdehyde (MDA) values, in vacuum-packaged chicken liver meat, both in the absence or presence of thyme essential oil (EO) during the entire storage period. Of the color parameters (L*, a*, b*) monitored during storage time, L* (lightness) values for vacuum-packaged chicken liver, irrespective of treatment, showed a varying trend, whereas a* (redness) values for V, VT1, and VT2 liver samples decreased, showing no significant differences. As determined by sensory analysis the observed shelf life of chicken liver samples was longest for VT2 (>12 d) followed by VT1 (12 d) and control (V) samples (7 d). Our results suggest that a Brochothrix thermosphact count (7 log CFU/g) and appearance of visible colonies can be used as indicators of chicken liver spoilage.