Biofilm formation of the food spoiler Brochothrix thermosphacta on different industrial surface materials using a biofilm reactor.

Brochothrix thermosphacta is considered as a major food spoiler bacteria. This study evaluates biofilm formation by B. thermosphacta CD337(2) - a strong biofilm producer strain - on three food industry materials (polycarbonate (PC), polystyrene (PS), and stainless steel (SS)). Biofilms were continuously grown under flow at 25 °C in BHI broth in a modified CDC biofilm reactor. Bacterial cells were enumerated by plate counting, and biofilm spatial organization was deciphered by combining confocal laser scanning microscopy and image analysis. The biofilms had the same growth kinetics on all three materials and reach 8log CFU/cm2 as maximal concentration. Highly structured biofilms were observed on PC and PS, but less structured ones on SS. This difference was confirmed by structural quantification analysis using the image analysis software tool BiofilmQ. Biofilm on SS show less roughness, density, thickness and volume. The biofilm 3D structure seemed to be related to the coupon topography and roughness. The materials used in this study do not affect biofilm growth. However, their roughness and topography affect the biofilm architecture, which could influence biofilm behaviour.

Characterization and interactions of spoilage of Pseudomonas fragi C6 and Brochothrix thermosphacta S5 in chilled pork based on LC-MS/MS and screening of potential spoilage biomarkers.

Pseudomonas and Brochothrix are the main spoilage organisms in pork, and each of these plays an essential role in the spoilage process. However, the effect of co-contamination of these two organisms in pork has not been elucidated. The changing bacterial communities during spontaneous spoilage of pork at 4 °C were evaluated using high-throughput sequencing. The dominant spoilage bacteria were isolated and these were identified as Pseudomonas fragi C6 and Brochothrix thermosphacta S5. Chilled pork was then experimentally contaminated with these strains, individually and in combination, and the progression of spoilage was assessed by analyzing various physicochemical indicators. These included total viable counts (TVC), pH, color, total volatile basic nitrogen (TVB-N), and detection of microbial metabolites. After 7 days of chilled storage, co-contaminated pork produced higher TVC and TVB-N values than mono-contaminated samples. Metabolomic analysis identified a total of 8,084 metabolites in all three groups combined. Differential metabolites were identified, which were involved in 38 metabolic pathways. Among these pathways, the biosynthesis of alkaloids derived from purine and histidine was identified as an important pathway related to spoilage. Specifically, histidine, histamine, AMP, IMP, GMP, succinic acid, and oxoglutaric acid were identified as potential spoilage biomarkers. The study showed that the combined presence of P. fragi C6 and B. thermosphacta S5 bacteria makes chilled pork more prone to spoilage, compared to their individual presence. This study provides insights that can assist in applying appropriate techniques to maintain quality and safety changes in meat during storage and further the assessment of freshness.

Isolation and characterization of Brochothrix phage ADU4.

B. thermosphacta is a psychrotrophic bacterium that often forms the predominant part of the spoilage microflora of aerobically and anaerobically stored meats. Bacteriophages are natural enemies of bacteria and their potential for use in environmentally friendly biocontrol of specific pathogens in food is being intensively studied. In this study, we reported the isolation and characterization of the newly isolated lytic Brochothrix phage ADU4, which is capable of infecting the B. thermosphacta bacterium. For the characterization of Brochothrix phage ADU4; host range, multiplicity of infection values (MOI), phage growth parameters (latent period and burst size), stability at various temperatures and pH, reduction growth of bacteria, effect on biofilm, and molecular characterization were investigated. The spot-test analysis showed positivity with B. thermosphacta strains, while no infection was observed in any other species and genera of bacteria tested. The optimal MOI value of the phage was determined as 0.1. The phage latent period and burst sizes were 40-50 min and 311 PFU/ml per infected host cell, respectively by one-step growth curve analysis. Brochothrix phage ADU4 reduced bacteria immediately after infection, which is shown by optical density (OD) measurement and colony counting (<10 CFU/ml) for 3 days. The degradation of B. thermosphacta in biofilm by Brochothrix phage ADU4 was analyzed and it was found that high titer phage breakdown the existing biofilm and also persistently inhibited biofilm formation. Brochothrix phage ADU4 genome was found to be 127,819 bp, and GC content 41.65%. The genome contains 217 putative open reading frames (ORFs), 4 tRNAs, and additionally, no known virulence and antibiotic resistance genes (AMR) were identified. Brochothrix phage ADU4 showed a high identity (96.09%) to the A9 phage that belongs to the Herelleviridae family. Nevertheless, the assembly module and its around appeared less conserved, and some DNA fragments in Brochothrix phage ADU4 genome were not found in A9 genome and vice versa. A9 contains TnpB, a transposase accessory protein involved in lysogenicity which is absent in Brochothrix phage ADU4. In contrary Brochothrix phage ADU4 had auxiliary metabolic genes (AMG) mostly carried by lytic phages. All these results showed that the Brochothrix phage ADU4 has excellent properties such as strong antibacterial activity, short latent period, high burst size, stability in different conditions, inhibition of biofilms, and absence of virulence and AMR genes. Based on all these features, this newly isolated phage is promising to control B. thermosphacta contamination in meat and meat products, and has the potential to be used alone or in combination with phage cocktails.

Antimicrobial mechanism of linalool against Brochothrix thermosphacta and its application on chilled beef.

This work aimed to explore the antibacterial ability and potential mechanism of linalool against Brochothrix thermosphacta (B. thermosphacta), providing knowledge of the preservation of chilled beef with linalool. The results found that linalool had an encouraging inhibitory effect on B. thermosphacta with a minimum inhibitory concentration (MIC) of 1.5 mL/L. Results of FESEM and zeta potential combined with probe labeling confirmed that linalool destroyed the cell structure thereby causing the leakage of intracellular components (AKP, protein, nucleic acid and ion). In addition, linalool caused respiratory disturbance by measuring the key enzyme activities including PK, SDH, MDH and ATPase. Energy limitation also appeared under linalool stress as seen from changes in ATP content (decreased by 56.06% and 69.24% in MIC and 2MIC groups, respectively). The respiratory inhibition rate of linalool to B. thermosphacta was 23.58% and the superposing rate with malonic acid was minimal (35.52%), suggesting that respiratory depression was mainly caused by the TCA cycle. Furthermore, accumulation of ROS and increase in MDA content (increased by 71.17% and 78.03% in MIC and 2MIC groups, respectively) accompanied by decreased activities of detoxification enzymes CAT and POD suggested that oxidative stress contributed to the bactericidal mechanism. Finally, linalool has been shown to effectively inhibit quality deterioration of chilled beef during storage by measuring pH, TVB-N and TVC without affecting sensory acceptability. All these highlight the great promise of using linalool as natural preservative for food industry.

Metabolomics reveals spoilage characteristics and interaction of Pseudomonas lundensis and Brochothrix thermosphacta in refrigerated beef.

Pseudomonas lundensis and Brochothrix thermosphacta are key spoilage microorganisms in aerobically stored chilled meat. The present study aimed to investigate the physicochemical and metabolomic profiles of refrigerated ground beef inoculated P. lundensis (PL) and B. thermosphacta (BT) as mono- or co-culture (BP). P. lundensis was the dominant spoilage strain in the co-culture of ground beef. A large amount of TCA-soluble peptide, TVB-N and TBA were formed in the PL and BP, while acetion was mainly produced in the BT, as accompanied by the different sensory and color changes. Meat metabolome indicated that 95, 396, and 409 metabolites with significant differences, were identified in ground beef inoculated BT, PL, and BP, respectively. These differential metabolites covered 58 metabolic pathways, in which histidine metabolism was identified as an important pathway related to spoilage in the three groups. Specifically, creatine, inosine, anserine, uracil, alanine, glutamine, 3-methylhistidine and 3-hydroxycapric acid were enriched as potential spoilage biomarkers. Taken together, those findings reveal the complex and competitive interactions of their co-culture of B. thermosphacta and P. lundensis, which provided a comprehensive insight into microbial spoilage mechanism in chilled beef.

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.

Suitability of lactic acid bacteria and deriving antibacterial preparations to enhance shelf-life and consumer safety of emulsion type sausages.

Ready-to-eat (RTE) sliced emulsion type sausages are sensitive to recontamination with Listeria (L.) monocytogenes during processing and packaging steps. Since Listeria spp. are able to grow on those products under cold storage conditions, taking steps to reduce the recontamination risk and implementing antibacterial hurdles contribute to consumer safety and increase the product quality. With this study data about the suitability of culture broth, cell-free supernatant (CFS) or concentrated bacteriocin preparations (CFSconc) of bacteriocin-producing lactic acid bacteria (LAB) obtained from fermented sausages from Germany as protective culture or antibacterial additive were provided. In different challenge tests, the potential of selected LAB or their preparations were investigated for their potential to reduce growth of L. monocytogenes and/or Brochothrix (B.) thermosphacta on sliced RTE emulsion type sausages under modified atmosphere or vacuum during refrigerated storage for a 21-day period. Applied LAB culture broth and CFS could not reduce the growth of L. monocytogenes or B. thermosphacta. On the other hand, samples treated with CFSconc obtained from Pediococcus spp. strains showed a significant inhibition (p < 0.05) of more than 1.5 log10 of the applied L. monocytogenes strains during the storage period. The growth of B. thermosphacta could not be influenced. Thereby, the need for concentrating preparations was shown to be important to obtain a suitable antibacterial preparation that would contribute to consumer safety and food quality when applied as a protective additive.

Development of two specific multiplex qPCRs to determine amounts of Pseudomonas, Enterobacteriaceae, Brochothrix thermosphacta and Staphylococcus in meat and heat-treated meat products.

Culturing methods are conventionally applied to investigate the contamination of food with several microorganisms after heat processing. However, with these methods, it is not possible to evaluate whether heat-treated meat products, such as cooked sausages, contained parts of spoiled meat. Therefore, two specific multiplex qPCRs were developed in this study in order to determine the microbiological quality of the raw materials used for these products. The PCR targets focused on four bacterial groups often found on meat (family Enterobacteriaceae, genus Pseudomonas, genus Staphylococcus and species Brochothrix thermosphacta). Specificity as well as sensitivity of the developed multiplex qPCRs, validated by using 68 microbial species, were 100%. The applicability of both multiplex qPCRs compared to culturing methods was performed using 96 meat samples (fresh and naturally spoiled) and 12 inhouse-made "Lyoner" sausages containing variable ratios of spoiled meat (0%, 5%, 12% and 25%; n = 3 for each group). Both methods showed similar results by evaluating the ∆log10 cfu/g, the relative accuracy and the t-test analysis (p > 0.05). Comparing qPCR results of the different sausage groups, a significant difference between sausages containing fresh meat and sausages containing spoiled meat (12% and 25%) was found only for Pseudomonas and B. thermosphacta in both raw and cooked sausages. The statistical difference between 5% vs. 12% and 25% spoiled meat in cooked sausages, was also found only for these two bacterial groups. The developed multiplex qPCRs were further applied to 30 commercially available "Bologna-type" sausages. The results showed a total of 14 sausages considered to be suspicious for Food Fraud. While the role of Staphylococcus spp. in meat spoilage remains unclear, Pseudomonas, Enterobacteriaceae and B. thermosphacta could together be used as an indicator for "spoiled meat" used in sausages. The developed qPCR systems in this study allow the detection of four relevant bacterial groups in the heated Bologna-type sausages and provide information about the hygienic quality of raw materials used. This method could thus be helpful for screening food suspected of Food Fraud.

Differentiation of meat-related microorganisms using paper-based surface-enhanced Raman spectroscopy combined with multivariate statistical analysis.

Surface-enhanced Raman spectroscopy (SERS) with subsequent chemometric evaluation was performed for the rapid and non-destructive differentiation of seven important meat-associated microorganisms, namely Brochothrix thermosphacta DSM 20171T, Pseudomonas fluorescens DSM 4358, Salmonella enterica subsp. enterica sv. Enteritidis DSM 14221, Listeria monocytogenes DSM 19094, Micrococcus luteus DSM 20030T, Escherichia coli HB101 and Bacillus thuringiensis sv. israelensis DSM 5724. A simple method for collecting spectra from commercial paper-based SERS substrates without any laborious pre-treatments was used. In order to prepare the spectroscopic data for classification at genera level with a subsequent chemometric evaluation consisting of principal component analysis and discriminant analysis, a data pre-processing method with spike correction and sum normalisation was performed. Because of the spike correction rather than exclusion, and therefore the use of a balanced data set, the multivariate analysis of the data is significantly resilient and meaningful. The analysis showed that the differentiation of meat-associated microorganisms and thereby the detection of important meat-related pathogenic bacteria was successful on genera level and a cross-validation as well as a classification of ungrouped data showed promising results, with 99.5% and 97.5%, respectively.

Tigecycline in the treatment of fulminant Mycoplasma pneumoniae pneumonia non-responsive to azithromycin and fluoroquinolone: A case report.

RATIONALE: Fulminant macrolide-resistant Mycoplasma pneumoniae pneumonia (MPP) has seldom been reported, and cases of MPP usually show rapid improvement after fluoroquinolones or tetracyclines addition. The purpose of this case report is to highlight the importance of proper selection of antibiotics for treatment of severe MPP and increase awareness concerning the emergence of fluoroquinolone-resistant MPP. PATIENT CONCERNS: A case of severe life-threatening pneumonia in a 26-year-old man with high fever and cough was non-responsive to azithromycin and fluoroquinolones. DIAGNOSES: The patient was diagnosed with MPP based on the test results of bronchoalveolar lavage using real-time quantitative PCR method. INTERVENTIONS: Tigecycline was given to the patient after azithromycin and fluoroquinolones failed. OUTCOMES: The patients fever subsided within the first day of tigecycline therapy. He showed rapid symptom resolution and improvement in lung infiltration after 4 days of tigecycline therapy. LESSONS: The case suggests that fulminant MPP should be timely treated with proper antibiotics, and the possible emergence of fluoroquinolone-resistant MPP should be of concern.

Effect of glucose, pH and lactic acid on Carnobacterium maltaromaticum, Brochothrix thermosphacta and Serratia liquefaciens within a commercial heat-shrunk vacuum-package film.

Carnobacterium maltaromaticum, Brochothrix thermosphacta and Serratia liquefaciens are common spoilage organisms found within the microbiome of refrigerated vacuum-packaged (VP) beef. Extending and predicting VP beef shelf-life requires knowledge about how spoilage bacteria growth is influenced by environmental extrinsic and intrinsic factors. Multifactorial effects of pH, lactic acid (LA) and glucose on growth kinetics were quantified for C. maltaromaticum, B. thermosphacta and S. liquefaciens within a heat shrink-wrapped VP commercial film containing a simulated beef medium. LA, pH, and undissociated lactic acid (UDLA) significantly affected bacterial growth rate (p < 0.001), whereas 5.55 mM glucose produced a marginal effect. At 1.12 mM UDLA, growth rate and maximum population density decreased 20.9 and 3.5%, 56 and 7%, and 11 and 2% for C. maltaromaticum, B. thermosphacta, and S. liquefaciens, respectively.

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).

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.

Metatranscriptomic analysis of modified atmosphere packaged poultry meat enables prediction of Brochothrix thermosphacta and Carnobacterium divergens in situ metabolism.

In this study, in situ-expressed metabolic routes of Brochothrix (B.) thermosphacta and Carnobacterium (C.) divergens were evaluated based on a metatranscriptomic dataset from bacteria growing on MAP chicken meat (O2/CO2; N2/CO2). Both species exhibited no (C. divergens) or minor transcription regulation (B. thermosphacta) within their main metabolic routes in response to different atmospheres. Both employ pathways related to glucose and ribose. Gluconeogenesis from lipid-borne glycerol is active in the progressing lack of carbohydrates. Pyruvate fates in both species comprise lactate, ethanol, acetate, CO2, formate, C4-compounds and H2O2 (only B. thermosphacta). Both species express genes for a minimal aerobic respiratory chain, but do not possess the genetic setting for a functional citric acid cycle. While products of carbohydrate and glycerol metabolism display mild to medium sensorial off-characteristics, predicted end products of their amino acid metabolism comprise, e.g., isobutyrate and isovalerate (B. thermosphacta) or cadaverine and tyramine (C. divergens) as potent spoilage compounds.

Comparison of Stomaching versus Rinsing for Recovering Bacterial Communities from Rainbow Trout (Oncorhynchus mykiss) Fillets.

ABSTRACT: The use of high-throughput methods allows a better characterization of food-related bacterial communities. However, such methods require large amounts of high-quality bacterial DNA, which may be a challenge when dealing with a complex matrix that has a low concentration of bacteria, such as fresh fish fillets. Therefore, the choice of method used to recover bacteria from a food matrix in a cost-effective way is critical, yet little information is available on the performance of commonly used methods. We assessed the recovery capacity of two such methods: stomaching and mechanical rinsing. The efficiency of the methods was evaluated through quantitative recovery and compatibility with end-point quantitative PCR (qPCR). Fresh rainbow trout (Oncorhynchus mykiss) fillets were inoculated with a bacterial marker, Brochothrix thermosphacta, at different concentrations (7.52 to 1.52 log CFU/g). The fillets were processed by one of the two methods, and the recovery of the marker in the suspensions was assessed by plate counting and qPCR targeting B. thermosphacta-rpoC. The same analyses were performed on six noninoculated fresh fillets. Stomaching and mechanical rinsing allowed efficient and repeatable recovery of the bacterial communities from the 42 inoculated fillets. No significant differences in recovery ratios were observed between the marker enumerated in the inoculation suspensions and in the corresponding recovery suspensions after rinsing and stomaching. However, the stomaching method allowed too many particles to pass through the filters bag, making necessary a limiting supplementary filtration step. As a consequence, only the rinsing recovery method allowed proper PCR quantification of the inoculated B. thermosphacta. The mean recovered bacterial level of the fillets was approximately 3 log CFU/g. It seems more relevant and cost-effective to recover the endogenous bacterial microbiota of a fish fillet structure using the rinsing method rather than the stomaching method.

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.

Antimicrobial Activity and Proposed Action Mechanism of 3-Carene against Brochothrix thermosphacta and Pseudomonas fluorescens.

3-Carene is an antimicrobial monoterpene that occurs naturally in a variety of plants and has an ambiguous antibacterial mechanism against food-borne germs. The antibacterial effects and action mechanism of 3-carene against Gram-positive Brochothrix thermosphacta ACCC 03870 and Gram-negative Pseudomonas fluorescens ATCC 13525 were studied. Scanning electron microscopy (SEM) examination and leakage of alkaline phosphatase (AKP) verified that 3-carene caused more obvious damage to the morphology and wall structure of B. thermosphacta than P. fluorescens. The release of potassium ions and proteins, the reduction in membrane potential (MP), and fluorescein diacetate (FDA) staining further confirmed that the loss of the barrier function of the cell membrane and the leakage of cytoplasmic contents were due to the 3-carene treatment. Furthermore, the disorder of succinate dehydrogenase (SDH), malate dehydrogenase (MDH), pyruvate kinase (PK), and ATP content indicated that 3-carene could lead to metabolic dysfunction and inhibit energy synthesis. In addition, the results from the fluorescence analysis revealed that 3-carene could probably bind to bacterial DNA and affect the conformation and structure of genomic DNA. These results revealed that 3-carene had strong antibacterial activity against B. thermosphacta and P. fluorescens via membrane damage, bacterial metabolic perturbations, and genomic DNA structure disruption, interfering in cellular functions and even causing cell death.

Inactivation of Yersinia enterocolitica and Brochothrix thermosphacta on pork by UV-C irradiation.

Ultraviolet (UV) irradiation has gained interest as a decontamination method for food for several years. This study investigated how UV-C affected the microbial load of pork, inoculated with Yersinia (Y.) enterocolitica and Brochothrix (B.) thermosphacta. The initial effect as well as the effect after 1, 7 and 14 days of storage were investigated. Additionally, the meat quality parameters color, pH value, myoglobin redox form percentages and antioxidant capacity were analyzed. During storage, the bacterial load on pork was significantly reduced up to 1.2 log10 using doses of 408 or 2040 mJ/cm2. In contrast to this, in vitro experiments with bacterial suspensions showed that calculated UV doses of 16.16 and 19.30 mJ/cm2 resulted in a 3.0 log10 reduction of Y. enterocolitica and B. thermosphacta, respectively. The analyzed meat quality parameters were not influenced by UV-C treatment. Hence, UV-C light can reduce microbial surface contamination without negatively affecting meat quality.

Encapsulation of cinnamon oil in cyclodextrin nanosponges and their potential use for antimicrobial food packaging.

The main goal of this work is the encapsulation of cinnamon essential oil in cyclodextrin nanosponges and the assessment of their antimicrobial activity against foodborne pathogens. After nanosponge synthesis, a headspace-solid phase microextraction coupled to gas chromatography-mass spectrometry (HS-SPME-GC-MS) method was validated to quantify essential oil major compounds. Results showed that essential oil was successfully encapsulated in cyclodextrin nanosponges with α-NS and ß-NS being able to encapsulate higher essential oil amounts. Cinnamon essential oil, alone and encapsulated in nanosponges, proved to have antimicrobial activity against foodborne bacteria. Time-kill assays proved that the essential oil, alone or encapsulated, had a bacteriostatic effect against all bacteria tested, with the exception of Y. enterocolitica where a bactericidal action was observed. Furthermore, the controlled release achieved by its encapsulation, allowed cinnamon essential oil to be effective at a much lower concentration in culture medium than when solely dissolved in culture medium. Thus, the results described herein encourage the use of cyclodextrin nanosponges as encapsulating agents for active food packaging applications.

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.