Spoilage potential and sensory profile associated with bacteria isolated from cold-smoked salmon

Off-odours/flavours associated with cold-smoked salmon spoilage are due to the activity of microflora. This study evaluated the spoilage potential of nine bacterial groups (Shewanella putrefaciens, Brochothrix thermosphacta, Aeromonas spp., Lactobacillus alimentarius, Lactobacillus sake,Lactobacillus farciminis, Carnobacterium piscicola, Photobacterium phosphoreum and Serratia liquefaciens) isolated from cold-smoked salmon. Five different isolates from each group were inoculated into sterile cold-smoked salmon blocks, and chemical and sensory changes were studied after five weeks of storage in vacuum packs at 6°C. Bacterial growth was monitored weekly during the storage period. A sensory profile was assigned to each group. Principal component analysis allowed some bacterial species to be characterised by a specific odour, and correspondence factorial analysis discriminated among the species according to their spoilage potential. The bacteria mainly responsible for spoilage were L. sake, L. farciminis and B. thermosphacta, which produced sulphurous, acidic and rancid off-odours respectively. Some strains of S. liquefaciens produced rubbery, cheesy or acidic off-odours. Some P. phosphoreum isolates were characterised by an acidic effect.     

Sensory and physicochemical evolution of tropical cooked peeled shrimp inoculated by Brochothrix thermosphacta and Lactococcus piscium CNCM I-4031 during storage at 8°C

This study investigated the sensory quality and physicochemical evolution (pH, glucose, l-lactic acid, biogenic amine, free amino-acids and volatile compounds) during storage at 8 °C of cooked peeled shrimp inoculated with the specific spoilage bacteria Brochothrix thermosphacta alone or mixed with the protective strain Lactococcus piscium CNCM I-4031. Growth of both bacteria was monitored at regular intervals during storage by microbial counts and the thermal temperature gradient gel electrophoresis (TTGE) technique. Bacterial counts showed that L. piscium and B. thermosphacta inoculated at 7 log CFU/g and 3 log CFU/g were well adapted to shrimp, reaching a maximum level of 9 log CFU/g after 4 days and 10 days respectively. In mixed culture, the growth of B. thermosphacta was reduced by 3.2 ± 0.1 log CFU/g. The TTGE technique allowed monitoring the colonisation of the strains on the shrimp matrix and confirming the dominance of L. piscium in mixed culture throughout the experiment. Sensory analysis confirmed that B. thermosphacta spoiled the product after 11 days, when its cell number attained 8 log CFU/g with the emission of strong butter/caramel off-odours. This sensory profile could be linked to the production of 2,3 butanedione, cyclopentanol, 3-methylbutanol, 3-methylbutanal, 2-methylbutanal, 4-methyl-3-chloro-3-pentanol and ethanol, which were produced in more significant quantities in the B. thermosphacta batch than in the batches in which the protective strain was present. On the contrary, TVBN and TMA were not suitable as quality indicators for B. thermosphacta spoilage activity. In the products where the protective L. piscium strain was present, no adverse effect on sensory quality was noted by the sensory panels. Moreover, biogenic amine assessment did not show any histamine or tyramine production by this strain, underlining its safety profile. Both strains produced lactic acid (1850 mg/kg in L. piscium and B. thermosphacta batch on days 3 and 10 respectively; 3830 mg/kg on day 7 in mixed culture) and the pH decrease from 6.6 ± 0.0 to 5.9 ± 0.1 was similar in all batches. Lactic acid production or competition for free amino-acid was not involved in the inhibition mechanism; however rapid glucose consumption by L. piscium could partially explain the growth limitation of the spoilage micro-organism. This study demonstrated the spoilage characteristic of B. thermosphacta and the usefulness of L. piscium as a bioprotective culture for tropical cooked peeled shrimp without any adverse effect on the sensory quality of the product.     

Evaluation of Natural Antimicrobials on Typical Meat Spoilage Bacteria In Vitro and in Vacuum-Packed Pork Meat

ABSTRACT: The aim of this study was to investigate the inhibitory effect of natural antimicrobials on the growth of typical spoilage bacteria from marinated pork. Minimum inhibitory concentrations (MIC) of thymol, cinnamaldehyde, allyl isothiocyanate, citric acid, ascorbic acid, a rosemary extract, and a grapefruit seed extract against Lactobacillus algidus, Leuconostoc mesenteroides, Leuconostoc carnosum, Carnobacterium maltaromaticum, Carnobacterium divergens, Brochothrix thermosphacta, and Serratia proteamaculans were determined in a microplate assay. Combinations of antimicrobials were tested and several combinations showed synergistic effects in inhibiting bacterial growth. Single and combined antimicrobials were added to vacuum-packed pork meat to evaluate preserving effects. Antimicrobial concentrations of up to 10 times the MIC values showed no effect on total bacterial growth in vacuum packed pork meaning that although most antimicrobials inhibited the growth of spoilage bacteria in vitro, results from the microplate assay could not be transferred to the meat system. Most natural antimicrobials possess strong odor and flavor that limit their use as a food preservative. In conclusion, this study showed that the use of natural antimicrobials in meat products is limited and that bacterial quality and shelf life was not enhanced under the chosen conditions.     

Characterization of psychrotrophic bacterial communities in modified atmosphere-packed meat with terminal restriction fragment length polymorphism

Characterization of psychrotrophic lactic acid bacteria (LAB) and Brochothrix thermosphacta communities is needed to understand the microbial ecology of spoilage of modified atmosphere-packed (MAP) meats. To overcome the limitations of the currently used methods for the characterization of psychrotrophic bacterial communities in meat, we developed a culture-independent, 16S rRNA gene-targeted terminal restriction fragment length polymorphism (T-RFLP) method. An identification library consisting of 100 Gram-positive and 30 Gram-negative meat-associated bacterial strains was set up to identify the terminal restriction fragments derived from the communities. The taxonomic resolution level of the T-RFLP method was in between genus and species within the investigated LAB strains and within family and genus within the investigated Gram-negative strains. The established library was applied to identify the members of bacterial communities in MAP minced meat at the end of the shelf life. The T-RFLP results and plate counts on Man-Rogosa-Sharpe, Violet Red Bile Glucose, and Streptomycin sulfate thallium acetate actidione agars indicated that LAB and B. thermosphacta predominated in meat. The bacterial taxa associated with the T-RFLP results were compared to those identified among plate-grown LAB isolates by numerical ribopattern analysis. Both methods agreed that Leuconostoc spp. and Carnobacterium spp. prevailed in the LAB community in minced meat followed by Lactobacillus algidus, Lactococcus spp. and Weissella spp. Colony identification revealed that Leuconostoc gasicomitatum, L. gelidum, Carnobacterium divergens and C. maltaromaticum were the predominant LAB species. The T-RFLP results were shown to correlate with viable counts of Leuconostoc spp. and B. thermosphacta. The T-RFLP method was found to be a useful tool enabling rapid and high-throughput characterization of psychrotrophic bacteria prevailing in MAP meat.     

Spoilage-related microbiota associated with chilled beef stored in air or vacuum pack

In order to study the spoilage-related microbiota of beef at species level, a combination of culture-independent and culture-dependent methods was used to analyse nine different beef samples stored at 4 °C in air or in vacuum pack. Plate counts on selective agars after 0, 7 and 20 days of storage showed that vacuum packaging reduced the viable counts of Brochothrix thermosphacta, Pseudomonas spp. and Enterobacteriaceae, whereas the growth of lactic acid bacteria (LAB) was unaffected. Storage in vacuum pack mainly affected viable counts and not necessarily the species diversity of microbial populations on meat. Such populations were studied by PCR-DGGE of DNA directly extracted from meat and from bulk cells from culture media, followed by sequencing of DGGE fragments. Pseudomonas spp., Carnobacterium divergens, B. thermosphacta, Rahnella spp. and Serratia grimesii, or close relatives were detected in the meat at time zero. The use of the culture-independent method highlighted the occurrence of species that were not detected by plating. Photobacterium spp. occurred in most meat samples stored in air or in vacuum pack, which indicates this organism probably has a role in spoilage. In contrast, culture-dependent analysis allowed detection of bacterial species that were not found in DNA extracted directly from meat. This was the case for several species of Serratia or Rhanella among the enterobacteria, and Leuconostoc spp. among the LAB. Besides advancing our knowledge of the species involved in the spoilage of vacuum-packaged meat, this study shows the benefits of combining culture-based and direct approaches to enhance understanding of populations of spoilage bacteria.     

Technology-induced selection towards the spoilage microbiota of artisan-type cooked ham packed under modified atmosphere

The microbiota associated with a highly-perishable Belgian artisan-type cooked ham was analyzed through plating and (GTG)₅-fingerprinting of isolates throughout its processing chain. The raw tumbled meat was characterized by the presence of a versatile microbiota around 4.8 log(cfu g⁻¹), consisting of lactic acid bacteria, staphylococci, Brochothrix thermosphacta, Gram-negative bacteria, and yeasts. Pasteurisation of the ham logs reduced bacterial counts below 2 log(cfu g⁻¹) and subsequent manipulations selected for leuconostocs and carnobacteria. Also, B. thermosphacta and several Enterobacteriaceae were found at this stage. During storage in an intermediate high-care area for 2 days, a selection towards certain Enterobacteriaceae (Hafnia alvei, Enterobacter spp., and Pantoea agglomerans) and lactic acid bacteria (mainly vagococci and Streptococcus parauberis) was observed. B. thermosphacta, Leuconostoc carnosum and carnobacteria were also detected, but only after allowing bacterial outgrowth by incubating the meat logs at 7 °C for four weeks. After a mild post-pasteurisation process and subsequent handling, incubation of the meat logs at 7 °C for four weeks led to outgrowth of Enterobacteriaceae (mainly Enterobacter spp. and Serratia spp.). B. thermosphacta, and lactic acid bacteria (Enterococcus faecalis, Leuc. carnosum, and Carnobacterium maltaromaticum) were also found. After slicing and packaging under modified atmosphere, the microbiota of the refrigerated end-product consisted of leuconostocs, carnobacteria, and B. thermosphacta.     

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.     

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/cm² was recorded after six weeks at 0, 2 and 7 °C, and ~ 3 log₁₀ CFU/cm² 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 16 s 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.     

Short communication: impact of pH and temperature on the acidifying activity of Carnobacterium maltaromaticum

The acidifying activity of Carnobacterium maltaromaticum LMA28, a strain isolated from French soft cheese, was studied in trypticase soy broth with yeast extract (TSB-YE) medium and in milk. In TSB-YE supplemented with lactose, glucose, or galactose, lactose and glucose were metabolized with a maximum growth rate of 0.32 h⁻¹ and galactose was not metabolized. During hydrolysis of lactose, the galactose moiety was not excreted. The major product was l(+) lactic acid, with no significant difference in the lactic acid yield. Glucose was not completely metabolized because cell growth stopped when pH values reached an average of 5.0. In sterilized UHT milk, the addition of 1 g/L of YE enhanced its coagulation. Compared with commercial starter lactic acid bacteria such as Lactococcus lactis DSMZ 20481 or Streptococcus thermophilus INRA 302, Carnobacterium maltaromaticum LMA 28 was shown to be a slow acidifying strain. However, in spite of this weak acidifying ability, C. maltaromaticum LMA 28 can sustain low pH values in coculture with Lc. lactis DSMZ 20481 or S. thermophilus INRA 302. The individual and interactive effects of initial pH values (5.2 to 8.0) and incubation temperatures (23 to 37°C) on acidifying activity were studied by response surface methodology. The 3 strains displayed different behaviors depending on pH and temperature. The psychrotrophic lactic acid strain C. maltaromaticum LMA 28 was able to grow at alkaline pH values and during storage conditions. It could be used as a potential ripening flora in soft cheese.     

Characterisation of the spoilage microbiota in raw salmon (Salmo salar) steaks stored under vacuum or modified atmosphere packaging combining conventional methods and PCR-TTGE

In order to characterise the spoilage related to microbiota of raw salmon, a combination of culture-dependent and -independent methods, including PCR–TTGE, was used to analyse 3 raw salmon batches stored for 3 days at chilled temperature in modified atmosphere packaging (MAP) (50% CO₂/50% N₂) or under vacuum. Sensory evaluation, microbiological enumeration and chemical analysis were performed after 3, 7 and 10 days of storage. At the onset of spoilage, 65 bacterial isolates were picked from the plates. Thus, 13 different genera or species were identified by phenotypic and molecular tests: Serratia spp., Photobacterium phosphoreum, Yersinia intermedia, Hafnia alvei, Buttiauxella gaviniae, Pseudomonas sp., Carnobacterium maltaromaticum, Carnobacterium divergens, Lactococcus piscium, Lactobacillus fuchuensis, Vagococcus carniphilus, Leuconostoc gasicomitatum and Brochothrix thermosphacta. The PCR–TTGE profiles and band identification enabled a shift of the dominant populations during the storage to be visualised for all the batches, probably due to the temperature change and the packaging. At the beginning of storage, Pseudomonas sp. dominated the raw salmon microbiota while in the following days (7 and 10), P. phosphoreum and L. piscium were identified as the main bacterial groups. This study enhances the knowledge of MAP and vacuum-packed raw salmon spoilage microbiota.     

Incorporation of bacteriocin in plastic retains activity and inhibits surface growth of bacteria on meat

The bacteriocin, nisin, was incorporated into a polyethylene based plastic film and retained activity against the indicator bacteria Lactobacillus helveticus and Brochothrix thermosphacta . Beef carcass surface tissue sections (BCT) topically inoculated with the psychrotrophic spoilage bacterium B. thermosphacta were vacuum-packaged both with and without wrapping with the nisin impregnated plastic and held at 4° C. An initial reduction of 2 log₁₀cycles of B. thermosphacta was observed with nisin-impregnated wrapped BCT within the first 2 days of storage. After 20 days of refrigerated storage, B. thermosphacta populations from nisin impregnated plastic wrapped samples were significantly less than (P<0·05) control vacuum-packaged samples; log₁₀5·8 vs 7·2 cfu cm⁻²respectively. Temperature abuse was simulated by shifting inoculated packs from 4° C (after 2 days) to 12° C. Again, by 20 days, the B. thermosphacta populations of treated samples wrapped with nisin impregnated plastic were significantly less than (P<0·05) control vacuum-packaged samples; log₁₀3·6 vs 6·3 cfu cm⁻²respectively. This work highlights the potential for incorporating antimicrobial peptides with a wider and different range of inhibitory activity directly into plastics of different properties for use in controlling food spoilage as well as preservation to enhance product microbial safety.     

Use of Carnobacterium maltaromaticum cultures and hydroalcoholic extract of Lippia sidoides Cham. against Listeria monocytogenes in fish model systems

Minimally processed refrigerated ready-to-eat fishes may offer health risk of severe infection to susceptible individuals due to contamination by the psychrotolerant bacterium L. monocytogenes. In this work, inhibition of L. monocytogenes by a plant extract and lactic acid bacteria (LAB) was studied in model fish systems kept at 5 °C for 35 days. For that, fillets of tropical fish “surubim” (Pseudoplatystoma sp.) and hydroalcoholic extract of the plant Lippia sidoides Cham. (“alecrim pimenta”) were used. Fish peptone broth (FPB), “surubim” broth and “surubim” homogenate were inoculated with combinations of L. monocytogenes and bacteriocin-producing Carnobacterium maltaromaticum (C2 and A9b⁺) and non bacteriocin-producing C. maltaromaticum (A9b^-), in the presence or absence of extract of “alecrim pimenta” (EAP). In all model systems, monocultures of L. monocytogenes and carnobacteria reached final populations ≥ 10⁸ CFU/ml after 35 days, except for L. monocytogenes in “surubim” homogenate (10⁴ CFU/ml). In FPB, EAP alone and combined with cultures of LAB inhibited L. monocytogenes but carnobacteria without EAP were only weakly antilisterial. In “surubim” broth, EAP alone did not prevent L. monocytogenes growth but cultures of carnobacteria combined or not with EAP inhibited L. monocytogenes, with more pronounced effect being observed for C. maltaromaticum C2, which produced bacteriocin. In “surubim” homogenate, EAP alone and combined with cultures of C. maltaromaticum A9b⁻ and A9b⁺ were strongly inhibitory to L. monocytogenes, while C. maltaromaticum C2 with EAP caused transient inhibition of L. monocytogenes. No significant inhibition of L. monocytogenes was observed for carnobacteria in “surubim” homogenate without EAP. In conclusion, it was observed that the use of EAP and cultures of carnobacteria have potential to inhibit L. monocytogenes in fish systems and the applications should be carefully studied, considering the influence of food matrix.     

Potential of a simple HPLC-based approach for the identification of the spoilage status of minced beef stored at various temperatures and packaging systems

The shelf life of minced beef stored (i) aerobically, (ii) under modified atmosphere packaging (MAP), and (iii) under MAP with oregano essential oil (MAP/OEO) at 0, 5, 10, and 15 °C was investigated. The microbial association of meat and the temporal biochemical changes were monitored. Microbiological analyses, including total viable counts (TVC), Pseudomonas spp., Brochothrix thermosphacta, lactic acid bacteria, Enterobacteriaceae, and yeasts/moulds, were undertaken, in parallel with sensory assessment, pH measurement and HPLC analysis of the organic acid profiles. Spectral data collected by HPLC were subjected to statistical analysis, including principal component analysis (PCA) and factorial discriminant analysis (FDA). This revealed qualitative discrimination of the samples based on their spoilage status. Partial least squares regression (PLS-R) was used to evaluate quantitative predictions of TVC, Pseudomonas spp., Br. thermosphacta, lactic acid bacteria, Enterobacteriaceae, and yeasts/moulds. Overall, the HPLC analysis of organic acids, was found to be a potential method to evaluate the spoilage and microbial status of a meat sample regardless of the storage conditions. This could be a very useful tool for monitoring the quality of meat batches during transportation and storage in the meat food chain.     

4-己基间苯二酚对凡纳滨对虾3 种腐败菌的抑菌活性及虾品质的影响

为研究4-己基间苯二酚（4-hexylresorcinol,4-HR）的抑菌活性及对虾品质的影响,通过体外及接菌实验分别评价了其对来源于凡纳滨对虾的麦芽糖肉食杆菌（Carnobacterium maltaromaticum）、腐败希瓦氏菌（Shewanella putrefaciens）和杀鲑气单胞菌（Aeromonas salmonicida）的抑菌活性及其对凡纳滨对虾冷藏期间品质变化的影响。牛津杯法和抑菌生长曲线显示4-HR能够显著抑制这3?种腐败菌的生长。通过对比处理前后腐败希瓦氏菌的电导率和流式细胞术分析,发现其抑菌机理可能与增强细菌膜通透性有关。将0.25?mg/mL的4-HR溶液涂膜于凡纳滨对虾上,发现在冷藏期间（4?℃）凡纳滨对虾的黑变程度较低而白度值较高,表明0.25?mg/mL?4-HR能有效抑制凡纳滨对虾的黑变。但涂膜4-HR的凡纳滨对虾适冷菌总数和挥发性盐基氮值仅在贮藏的前4?d低于未涂膜的对照组;而到贮藏终点时,涂膜4-HR的凡纳滨对虾适冷菌总数较对照组高0.14（lg（CFU/g））。结果表明尽管4-HR对3?种腐败菌具有良好的体外抑菌活性,但在凡纳滨对虾贮藏过程中4-HR优先作为防黑变剂抑制对虾黑变。     

Influence of temperature, pH and NaCl concentration on the maximal growth rate of Brochothrix thermosphacta and a bioprotective bacteria Lactococcus piscium CNCM I-4031

The maximum specific growth rate (μ_max) of Brochothrix thermosphacta, a spoilage bacteria of cooked peeled shrimp, and Lactococcus piscium CNCM I-4031, a bioprotective strain, was investigated under different conditions of temperature, NaCl concentrations and pH. The basic modelling approach used was the Gamma concept (γ-concept) and the model developed was then adapted to shrimp. Cardinal growth parameters were quite similar for the two strains, except for NaCl. No NaCl was required for growth and the NaCl_max was three-times higher for B. thermosphacta than for L. piscium (62 and 23 g l⁻¹ respectively). However, tolerance to NaCl was higher in seafood than in liquid broth, possibly due to presence of osmoltically active molecules. L. piscium and B. thermosphacta were psychrotolerant, with T_min = −4.8 and −3.4 °C, T_opt = 23.4 and 27.0 °C and T_max = 27.2 and 30.8 °C respectively. The optimal pH was neutral and growth possible till pH = 4.8 for the two strains, assuming possible applications of the bioprotective strain in lightly marinated seafood. The μ_max of B. thermosphacta in shrimp was a little higher than in L. piscium whatever the environmental conditions. Validation of the model showed that the γ-concept was suitable for predicting μ_max of B. thermosphacta in shrimp. Data generated in this study can be used to adapt the model to other foods with few additional experiments and the effect of different parameters may be added in the future. The model was less accurate for the bioprotective strain and the effect of NaCl must be studied in more detail directly in the matrix.     

Protective action of Lactobacillus curvatus CRL705 on vacuum-packaged raw beef. Effect on sensory and structural characteristics

Lactobacillus curvatus CRL705 was examined for its effectiveness as protective culture in the biopreservation of vacuum-packaged fresh beef stored during 60 days at 2 °C. For this purpose, L. curvatus CRL705, producer of lactocin 705 and lactocin AL705, was inoculated on the meat surface (10⁶ cfu g⁻¹). This microorganism became the dominating population throughout the storage period controlling the growth of Brochothrix thermosphacta and spoilage lactic acid bacteria naturally present on the meat. When the microstructural characteristics of the meat were evaluated using light microscopy, beef samples inoculated with the bioprotective culture showed a 10 days delay for the appearance of tissue degradation signs. Sensory analysis demonstrated that beef samples treated with L. curvatus CRL705 only developed an “acid” off-flavor after 60 days of refrigerated storage, and no undesirable off-odors were found. Therefore, inoculation with this bacteriocinogenic strain would provide an additional hurdle to improve storage life of refrigerated vacuum-packaged beef without affecting its sensory and structural characteristics.     

Characterization of Carnobacterium maltaromaticum LMA 28 for its positive technological role in soft cheese making

Carnobacterium maltaromaticum is a lactic acid bacterium isolated from soft cheese. The objective of this work was to study its potential positive impact when used in cheese technology. Phenotypic and genotypic characterization of six strains of C. maltaromaticum showed that they belong to different phylogenetic groups. Although these strains lacked the ability to coagulate milk quickly, they were acidotolerant. They did not affect the coagulation capacity of starter lactic acid bacteria, Lactococcus lactis and Streptococcus thermophilus, used in dairy industry. The impact of C. maltaromaticum LMA 28 on bacterial flora of cheese revealed a significant decrease of Psychrobacter sp. concentration, which might be responsible for cheese aging phenomena. An experimental plan was carried out to unravel the mechanism of inhibition of Psychrobacter sp. and Listeria monocytogenes and possible interaction between various factors (cell concentration, NaCl, pH and incubation time). Cellular concentration of C. maltaromaticum LMA 28 was found to be the main factor involved in the inhibition of Psychrobacter sp. and L. monocytogenes.     

Real-Time Detection of Volatiles Released During Meat Spoilage: a Case Study of Modified Atmosphere-Packaged Chicken Breast Fillets Inoculated with <Emphasis Type="Italic">Br. thermosphacta</Emphasis>

The microbial spoilage of meat is accompanied by the release of volatile organic compounds (VOCs), many of which are odorous. These compounds give spoiled meat its characteristic pungent, sour, sulphury odour that provides consumers with an indication that the meat is unpalatable. Characterising meat spoilage based on volatile markers is of interest to the food industry in view of developing food freshness indicators (FFIs) to maintain food quality and reduce food waste. Conventional analytical methods for detecting VOCs developing during food spoilage involve intermittent sampling that delivers only snapshots of the release processes and thereby only limited information on the kinetics of their production and release. Proton-transfer-reaction mass spectrometry (PTR-MS) is an on-line technique that enables the detection of VOCs in real-time, thereby offering the possibility to follow the release of VOCs with a high time resolution. An analytical method using PTR-MS was developed to enable the continuous detection of VOCs released from chicken breast fillets inoculated with Brochothrix thermosphacta and stored under modified atmosphere (30 % CO₂, 70 % O₂) at 4 °C for 1 week. The meat spoilage VOCs detected by PTR-MS displayed different temporal dynamics of production and release, depending on the extent of spoilage. This paper describes the development of an analytical set-up for real-time detection of volatile spoilage markers using PTR-MS. This case study using inoculated chicken breast fillets demonstrates the applicability of this method to characterise individual VOC release patterns, which is of potential utility in the development of FFIs for the consumer market.     

Effect of storage time and heat processing on the volatile profile of Senegalese sole (Solea senegalensis Kaup, 1858) muscle

The effect of heat treatment and the presence or absence of fish skin on the volatile composition of Senegalese sole muscle was studied. The volatile profile of Senegalese sole at different storage periods was also evaluated. All samples were analysed by HS-SPME–GC–IT/MS and subjected to sensory evaluation. As expected, cooking enhanced the production/liberation of volatile compounds. Fish with the skin present, after cooking, had higher levels of sulphur compounds, 2-nonanone, ethyl octanoate and lower contents of hexanol and heptanol than skinned fish; moreover, the samples with the skin had a better overall sensory acceptability. During storage, changes on the volatile composition of Senegalese sole samples were found. The major differences were obtained after 2 weeks of storage. Compounds such as hexanal, heptanal, octanal, decanal, (E)-2-hexenal, (E)-2-decen-1-al, (E,Z)-2,6-nonadienal, benzaldehyde, 4-ethyl-benzaldehyde, 1-penten-3-ol, heptanol and (E)-2-octen-1-ol decreased after 2 weeks of storage, and other compounds, such as 3-methyl-1-butanal, 2-methyl-1-butanal, 2-heptanone, dimethyl trisulphide, dimethyl tetrasulphide and 2-methyltetrahydrothiophen-3-one increased. These differences were confirmed by sensory evaluation. Principal component analysis was applied to the chemical data.