Analysis of volatile compounds produced by different species of lactobacilli in rye sourdough using multiple headspace extraction

Profiles of volatile organic compound (VOC) produced by nine individual lactic acid bacteria (LAB) during rye sourdough fermentation were compared by automated SPME and GC/MS‐Tof. The dough samples were inoculated with individual strains, placed inside the headspace vials and incubated during next 24 h. The production or loss of VOC‐s was followed by adsorbing volatiles onto 85‐m Car/PDMS fibre in every 4 h. Volatile profiles differed among LAB species and divided LAB into two main groups – hetero‐ and homofermentative. Hetrofermentative LAB (Lactobacillus brevis; Leuconostoc citreum; Lactobacillus vaginalis, Lactobacillus panis) showed high production of acetic acid, CO₂, ethanol, ethylacetate, producing also hexyl acetate, ethyl hexanoate and isopentyl acetate. Whereas homofermentative LAB species (Lactobacillus helveticus; Lactobacillus casei; Lactobacillus sakei; Lactobacillus curvatus) produced a considerable amount of 2,3‐butanedione. Production of l ‐leucine methyl ester was unique for Lb. sakei, Lb. casei and Lb. curvatus strains. Lb. helveticus was the only LAB that produced benzaldehyde.     

Study of the Lactobacillus sakei protective effect towards spoilage bacteria in vacuum packed cooked ham analyzed by PCR-DGGE

The effectiveness of Lactobacillus sakei B-2 inoculated as a protective culture on the inhibition of spoilage bacteria on sliced vacuum packed cooked ham was investigated by using culture-dependent and -independent approaches. Total microbial DNA was directly extracted from both control and treatment samples, and subjected to a nested PCR protocol, PCR–DGGE analysis was used to identify and monitor the dynamic changes in the microbial population, followed by partial 16S rDNA sequencing. The DGGE profile demonstrated that the protective culture effectively suppressed growth of predominant spoilage bacteria L. sakei, Lactobacillus curvatus and Leuconostoc mesenteroides in cooked ham during storage at 4 °C, however, growth of uncultured Leuconostoc was not inhibited. The shelf-life of this product inoculated with L. sakei B-2, at levels of 5.91 ± 0.04 log₁₀ CFU g⁻¹ was 35 days, compared to 15 days of control samples, when the ham was stored at 4 °C.     

Survival of biogenic amine‐producing dairy LAB strains at pasteurisation conditions

Biogenic amines (BAs) are low molecular weight organic bases with biological activity, synthesised by the microbial decarboxylation of precursor amino acids such as histidine, tyrosine or arginine. The consumption of food and beverages containing large amounts of BA can have toxicological effects. This paper reports the resistance of aminogenic lactic acid bacteria strains to pasteurisation (63 °C for 30 min). Skimmed milk inoculated with histamine‐ and tyramine‐producing strains of Enterococcus and Lactobacillus was subjected to pasteurisation. The viable number of Enterococcus durans IPLA 655 was reduced by 1 log, while that of Enterococcus faecalis BA64 was reduced by 3 logs. Lactobacillus brevis CECT 3810 did not survive pasteurisation, while the other Lactobacillus strains examined were more resistant; their viable numbers were reduced by about 2 logs. The two most resistant strains, E. durans IPLA 655 and Lactobacillus curvatus VI6, were subjected to higher temperatures. Enterococcus durans was completely inactivated at 78 °C, while some live cells of Lb. curvatus were observed. These results show that some native, metabolically active BA producers could be found in pasteurised milk owing to their resistance to the thermal treatment; their presence could have a negative impact on the final content of BAs in dairy products made from such milk.     

Inhibition by Lactobacillus sakei of other species in the flora of vacuum packaged raw meats during prolonged storage

The abilities of five Lactobacillus sakei strains and one Lactococcus lactis strain to retain inhibitory activity against several target organisms in the flora of product during 12 weeks storage of vacuum-packaged lamb and beef was investigated. L. sakei strains were generally found capable of developing dominant populations on both beef and lamb. L. lactis 75 grew poorly on lamb did not inhibit co-inoculated Brochothrix thermosphacta. Lamb inoculated with the Sakacin-A producer L. sakei Lb706 had lower Listeria monocytogenes populations than lamb inoculated with a bacteriocin-negative variant. In beef packs inoculated with Clostridium estertheticum spores and L. sakei strain 27, 44 or 63, the development of blown-pack spoilage was delayed by up to one week. Campylobacter jejuni inoculated onto beef was recovered from fewer packs when it was co-inoculated with 3000 CFU cm⁻² of L. sakei strain 27, 44 or 63. Observed inhibition did not always correlate with inhibition observed in earlier media-based studies, supporting the view that functionality identified using simple media-based screening methods may not be replicated in the complex environment of stored foods, and vice-versa. These findings further define a set of L. sakei strains with potential for the extended bio-preservation of minimally processed fresh beef and lamb.     

Sensory, physical, chemical and microbiological changes in European sea bass (Dicentrarchus labrax) fillets packed under modified atmosphere/air or prepared from whole fish stored in ice

Quality changes and shelf‐life in European sea bass fillets packed under 40% CO₂: 60% N₂ (MAP) and air (AIR) or prepared from the whole ungutted fish stored in ice (ROUND) were compared. Raw and cooked sensory scores, pH, colour, expressible water, malonaldehyde, total viable count, Pseudomonas spp., H₂S‐producing bacteria, Lactobacillus spp., Streptococcus spp., Staphylococcus spp., coliforms and proteolytic counts, were determined until AIR and MAP raw fillet spoilage. Raw ROUND fillets had the best quality and shelf‐life (10 vs. 8 vs. 7 days after slaughtering in ROUND, MAP and AIR, respectively), while the MAP fillets had better sensorial scores, lower pH values and better microbiological counts, but greater lightness values than the corresponding AIR fillets. MAP fillets also had the highest malonaldehyde levels. The higher correlation between Streptococcus spp. and odour scores (r = ?0.971, P < 0.01) compared to the other species could suggest that it is a specific spoilage organism in the MAP condition used.     

Comparison of the shelf lifes of map and vacuum packaged hot smoked rainbow trout (Onchoryncus mykiss)

The objective of this study was to determine the differences of microbiological and chemical quality in MAP and VP packages in refrigerated smoked trout fillets. Such information is important in evaluating the safety of these products. Three groups were constituted: group A – packed in 60%Co₂/40% N₂; group B – packed in 50%CO₂/50% N₂ and group C– vacuum packed. The ratio of fillet to package volume was 1/3 chemical analysis and microbiological analysis were done and also instrumental and sensory analysis were used to determined the shelf life. The observed shelf life of smoked trout was found at least 33 days in vacuum package, 47 days in 60% CO₂/40% N₂ gas mixture and 40 days in 50% CO₂/50% N₂ gas mixture according to results of microbiological analysis. The observed shelf life of smoked trout in MAP group A was determined as 14 days more than VP packs according to results of microbiological and sensory analysis in the study.     

Bacteriocin-producing lactobacilli in Spanish-style fermented sausages: characterization of bacteriocins

Three bacteriocins were studied. Sakacin K, a bacteriocin from Lactobacillus sake CTC494 was purified to homogeneity by a four-step system involving ammonium sulphate precipitation, binding to a cation exchanger, hydrophobic interaction and reverse phase chromatography in FPLC (fast performance liquid chromatography) system. The peptide sequence was determined by Edman degradation. The first 30 amino acid residues from the N-terminus of sakacin K were identical to those of curvacin A from Lactobacillus curvatus LTH1174 and sakacin A from Lactobacillus sakei Lb706. The structural gene of sakacin K in Lb. sake CTC494 was located on a 60 Kbp plasmid (as has been previously reported) by curvacin A in Lb. curvatus LTH1174 and sakacin A in Lb. sakei Lb706. Plantaricin D, a bacteriocin-like compound isolated from Lb. plantarum CTC305 was purified and sequenced using the same procedure as described for sakacin K. The first 15 amino acid residues of plantaricin D were identical to those obtained from the bacteriocin inducer peptide (termed plantaricin A) of the bacteriocinogenic system in Lb. plantarum C11. The structural gene of the plantaricin D peptide was located on a similar EcoRl chromosomal fragment when compared to plantaricin A in Lb. plantarum C11. These two isolated bacteriocin-like peptides (sakacin K and plantaricin D) were purified from two new different strains obtained from fermented meat, confirming the ecological importance of these substances. Lactobacillus sakei CTC372, the third bacteriocinogenic strain selected in this study from fermented sausages, produced a bacteriocin named sakacin T. The bacteriocin is not found free in the supernatant of the cells. It is active against Listeria monocytogenes and Staphylococcus aureus. The genetic determinant of this bacteriocin was localized in a 84·4 Kbp plasmid by conjugative transfer and curing assays.     

Modelling contributes to the understanding of the different behaviour of bacteriocin-producing strains in a meat environment

Bacteriocins from lactic acid bacteria are potent antibacterial agents that enable the producer cells to selectively and efficiently inhibit a part of the competing background flora, including several spoilage bacteria and foodborne pathogens. The use of bacteriocinogenic lactic acid bacterium strains, either as starters or as protective cultures, is very limited in the food industry, possibly because many questions about the in situ functionality of such strains remain unanswered. A modelling approach was developed to study the functionality of lactic acid bacterium bacteriocin producers in a meat environment. The kinetic properties of three potential bacteriocin-producing starter cultures for sausage fermentation, namely Lactobacillus sakei CTC 494, Lactobacillus curvatus LTH 1174, and Lactobacillus amylovorus DCE 471 were compared in MRS, used as a meat simulation medium. Moreover, the application possibilities of the bacteriocin-producing Enterococcus faecium RZS C5 strain were evaluated. It turned out that growth and bacteriocin production by Lb. sakei CTC 494 and Lb. curvatus LTH 1174 are adapted to the meat environment, whereas Lb. amylovorus DCE 471 is kinetically not able to dominate the meat fermentation process. Furthermore, the use of bacteriocin-producing enterococci as functional cocultures seems to be attractive.     

Microbial and physicochemical succession in fermented sausages produced with bacteriocinogenic culture of Lactobacillus sakei and semi-purified bacteriocin mesenterocin Y

The influence of the bacteriocinogenic culture Lactobacillus sakei (10⁵/g) and semi-purified bacteriocin mesenterocin Y (2560 AU/kg) on the safety and quality of traditional Croatian fermented sausages was investigated. The addition of Lb. sakei and/or mesenterocin Y reduced microbial counts (P < 0.05) in the final products. After 28 days of ripening, coagulase-negative cocci decreased 1.5–2.0 log, yeasts 1.2–1.4 log and enterococci 1.7–2.7 log. In the case of the addition of Lb. sakei, the lactic acid bacteria count was significantly (P < 0.05) higher at day 7 of ripening, and was accompanied by a lower pH and a higher amount of lactic acid (P < 0.05). In the final product the amount of acetic acid was significantly lower. More intensive proteolysis and an increase in ammonia content were found at the beginning of fermentation, and in the second phase of ripening in the control samples, respectively. The free fatty acid concentration was significantly lower during the entire ripening process compared to the control (P < 0.05). Semi-purified mesenterocin Y did not affect the sensory properties of the sausages, whilst the addition of Lb. sakei enhanced them.     

Identification of odour‐active compounds in muscle of brown trout (Salmo trutta) as affected by dietary lipid sources

The effect of fatty acid composition on odour‐active compounds in brown trout (Salmo trutta) muscle was evaluated. The fillets were obtained from three groups of fish fed experimental diets containing either fish oil (FO), soybean oil (SO) or linseed oil (LO). Muscle fatty acid composition was shown to be influenced by diet. Thirty‐one odorous compounds were detected by gas chromatography/olfactometry (frequency‐of‐detection method). Most of these compounds were formed by the oxidation of unsaturated fatty acids. Independently of diet, (E)‐2‐pentenal, (E)‐2‐pentenol and (E)‐2‐hexenol contribute strongly to the odour of brown trout. (E,Z)‐2,4‐Heptadienal was detected with high frequency in fish fed diets containing high levels of n‐3 PUFAs (FO and LO groups). Hexanal, (E)‐2‐hexenal and 2‐nonanol seem to contribute most to the odour of fish fed diets containing vegetable oils. Many odorous compounds were derived from the oxidation of mono‐ and di‐unsaturated fatty acids, which could be promoted by high levels of PUFAs. © 2002 Society of Chemical Industry     

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.     

EFFECT OF FILMS OF DIFFERENT OXYGEN TRANSMISSION RATE ON TOXIN PRODUCTION BY CLOSTRIDIUM BOTULINUM TYPE E IN VACUUM PACKAGED COLD AND HOT SMOKED TROUT FILLETS

Studies were done to determine the effect of film oxygen transmission rate (OTR) on the time to toxicity in vacuum packaged cold and hot smoked rainbow trout fillets challenged with C. botulinum type E (10² spores/g) and stored at refrigerated conditions (4C), and under mild (8C) and moderate (12C) temperature abuse conditions. While no samples were toxic at 4C, toxin was detected within 28 days at 8C for cold smoked trout fillets vacuum packaged in films with high OTR. Toxin was also detected for most vacuum packaged hot smoked trout fillets within 14–28 days at 8C, with the exception of trout fillets packaged in films with an OTR > 10,000 cc/m²/day. In most cases at 8C, spoilage, based on odor/color scores, preceded or occurred simultaneously with toxigenesis. At 12C all cold and hot smoked trout were toxic after 14–21 days and samples packaged in films with an OTR <5000 cc/m²/day became toxic before, or at the same time as, samples became spoiled. This study has shown that vacuum packaging of trout fillets in low gas barrier films, ranging in OTR from approximately 3,000 to approximately 10,000 cc/m²/day at 24C and 0% relative humidity (RH), did not prevent the growth and toxin production by C. botulinum in vacuum packaged cold and hot smoked trout fillets at 12C. Additional barriers, other than the OTR of the packaging film, need to be considered to ensure the safety of vacuum packaged trout fillets, particularly at mild to moderate temperature abuse storage conditions.     

Effect of the lactic acid bacteria on the control of listerial activity and shelf life of smoked salmon scraps

The potential protective activity of lactic acid bacteria (LAB) from cold smoked salmon scraps was evaluated towards Listeria monocytogenes. Seventy‐three LAB strains were isolated and identified by biomolecular methods; Lactobacillus curvatus and Lactobacillus sakei prevailed. Three of the strains tested, identified as L. sakei, profile O, had a significant inhibitory activity against L. monocytogenes ATCC 19115 and clones DUP‐1042 and DUP‐18596. The evolution of microbial populations and chemical parameters were determined at time intervals to verify the shelf life. Listeria monocytogenes was isolated in half the packages also exceeding the legal limit. The shelf life of scraps was set at 30 days. Clonal characterisation of L. monocytogenes was performed by ribotyping. DUP‐1042, one of the human pathogen clones, was the most represented pattern. The results suggest further studies aimed at the selection of autochthonous nonspoilage LAB strains as bioprotective agents for cold smoked salmon.     

Spoilage Processes and Proteolysis in Chicken as Detected by HPLC

Fresh poultry fillets inoculated or not with Pseudomonas fragi stored under aerobic, vacuum and 100% CO₂ packaging conditions to study the microbiological as well as the changes occurring in the concentration of glucose, lactate and water-soluble proteins, during the storage at 3 and 10°C. It was found that lactic acid bacteria were the dominant organisms in inoculated or uninoculated poultry fillets stored under these conditions. The low-molecular-weight compounds such as glucose and L -lactate were found to decrease always in these fillets during storage and the profile of water-soluble proteins as detected by HPLC differed among the different samples treated differently. © 1997 SCI.     

Production of organic acids by Lactobacillus strains in three different media

Ten strains of Lactobacillus (Lb). casei, Lb. rhamnosus, Lb. plantarum, Lb. paracasei and Lb. curvatus species were chosen to determine the production of organic acids after cultivation in skimmed milk, MRS broth and Jerusalem artichoke (JA) medium. The highest acidity was obtained in MRS broth and the weakest acidification was found in skimmed milk. Lb. casei Shirota produced the highest amount and Lb. rhamnosus VT1 the lowest amount of substances being estimated as titratable acidity. All strains produced lactic acid in the investigated broth and most of the strains produced acetic acid in MRS broth except Lb. curvatus 2768 and Lb. casei Shirota, in JA broth except Lb. paracasei SF1 and in skimmed milk except Lb. casei 2750, Lb. curvatus 2768, Lb. curvatus 2775 and Lb. casei Shirota. All strains, except Lb. plantarum 01, produced butyric acid in MRS broth. Beside the lactic and acetic acids, formic, citric, succinic and glutamic acids were also produced in MRS broth; formic and succinic acids were produced in skimmed milk and succinic acid in JA broth. Some strains showed change in their fermentation profile from homofermentative to mix-acid fermentation in milk. The antifungal efficiency of the lactic and acetic acid in the amount produced by lactobacilli was investigated. None of the investigated aspergilli were inhibited. The inhibitory effect of acids against Fusarium increased unequivocally with the increasing concentration. The study pointed at the dissimilarity of organic acid production of Lactobacillus strains, which was considerably influenced by the media.     

Probiotic monocultures in fermented goat milk beverages – sensory quality of final product

The aim of our study was to conduct a selection of the monocultures capable of providing the most attractive sensory features of the final product. Four fermented goat's milk beverages were produced with probiotic monocultures containing Lactobacillus (Lb. acidophilus La‐5, Lb. rhamnosus K3 and Lb. plantarum O20) and Bifidobacterium (Bif. animalis subsp. lactisBB‐12). A sensory analysis and microbiological assessment of fermented goat's milk beverages were made at the beginning of the study and after 3, 7, 10 and 14 days of refrigerated storage (5 ± 1 °C). We found that samples including monocultures Lb. plantarum O20 and Bif. animalis subsp. lactisBB‐12 were differentiated from other goat's milk beverages.     

Molecular Characterization of Lactic Acid Bacteria in Levačka Sausage

Levačka sausage is traditionally fermented dry sausage which is produced in central Serbia. It is made of beef and pork with the addition of solid fat and natural spices. The whole manufacturing process lasted for 21 days. The goal of this study was to create a collection of lactic acid bacteria isolated during the fermentation and identify them using molecular methods. A total of 50 isolates from different stages of fermentation were identified by molecular methods. Pediococcus pentosaceus, Ln. mesenteroides were predominant microorganisms in levačka sausage. Lb. curvatus, Lb. sakei and Lb. carnosum were also identified.     

The effect of a flaxseed oil-enhanced diet on the product quality of farmed brook trout (Salvelinus fontinalis) fillets

Abstract: The effects of dietary modification with flaxseed oil‐enhanced (Flax) feed on the product quality of brook trout fillets were examined. Trout were fed a commercial feed supplemented with fish oil (CD) or flaxseed oil (Flax) for 165 d before harvesting. Proximate composition and fatty acid profile were determined on fillets. Quality parameters of the raw fillets were examined over the storage period by measuring color (L*, a*, b*), muscle pH, and thiobarbituric acid reactive substances test. Evaluations on the cooked fillets included sensory evaluation with triangle tests and a paired preference test. There were no differences in proximate composition between the groups; however, the total omega‐3 fatty acids were greater in Flax fillets (P < 0.05). Diet and day were shown to interact in their effect on whiteness, pH, and lipid oxidation (P < 0.05); however, linear regression did not determine that malondialdehyde concentration was associated with time in either diet type implying that lipid oxidation in the vacuum‐packed fish was controlled at storage temperatures (4°C). Sensory panelists were able to choose the odd sample in a replicated triangle test analyzed using the β‐binomial model, and there was preference for Flax fillets (P < 0.05). Results indicate that a Flax‐enhanced diet would have favorable effects on product quality of farmed brook trout.     

Combined effect of an oxygen absorber and oregano essential oil on shelf life extension of rainbow trout fillets stored at 4 °C

In the present study the combined effect of an O₂ absorber and oregano essential oil (0.4% v/w) on shelf life extension of rainbow trout fillets (Onchorynchus mykiss) stored under refrigeration (4 °C) was investigated. The study was based on microbiological [TVC, Pseudomonas spp., Lactic Acid Bacteria, H₂S-producing bacteria including Shewanella putrefaciens, Enterobacteriaceae and Clostridium spp.), physicochemical (pH, PV, TBA, TVBN and Drip loss) and sensory (odor, taste) changes occurring in the product as a function of treatment and storage time. Aerobically-packaged rainbow trout fillets stored at 4 °C were taken as control samples. Results showed that TVC exceeded 7 log cfu/g on day 4 of storage for control samples, day 7–8 for samples containing oregano oil, day 9 for samples containing the O₂ absorber and day 12–13 for samples containing the O₂ absorber and oregano oil. Pseudomonas spp., Enterobacteriaceae and LAB were only partially inhibited by the O₂ absorber and/or the oregano oil. In all cases the inhibition effect was more pronounced when the combination of O₂ absorber with oregano essential oil was used. pH decreased from an initial value of 6.65–6.09 and subsequently increased to 6.86 due to formation of protein decomposition products. % Drip loss ranged between 7% and 11–12% at the end of the product shelf life. PV values ranged between 11.4 and 27.0 meq O₂/kg oil while malondialdehyde (MDA) ranged between 9.6 and 24.5 mg/kg. TVBN ranged between 10.6 and 54.6 mg/kg at the time of sensory rejection. Sensory shelf life was 4 days for the control samples, 7–8 days for samples containing oregano oil, 13–14 days for samples containing the O₂ absorber and 17 days for samples containing the O₂ absorber plus oregano oil.     

Inhibition of bacterial growth on ham and bologna by lysozyme,nisin and EDTA

Ham and bologna sausages were prepared with or without addition of 500 mg kg⁻¹ lysozyme:nisin, 1:3, and 500 mg. kg⁻¹ EDTA. Sausages were inoculated with one of; Brochothrix thermosphacta, Escherichia coli O157:H7, Lactobacillus sakei, Lactobacillus curvatus, Leuconostoc mesenteroides, Listeria monocytogenes, Salmonella typhimurium, Serratia grimesii or Shewanella putrefaciens, vacuum packed and stored for 4 weeks at 8°C. Plate counts were made on selective and nonselective media. Inhibitor treatment reduced initial populations of B. thermosphacta and Lc. mesenteroides on both meats. Treatment of ham and bologna prevented growth of B. thermosphacta, to week 4. Treatment reduced growth of Lb. curvatus on ham and bologna, to week 3. Treatment of bologna reduced growth of Lc. mesenteroides and L. monocytogenes for 2 weeks. Treatment of ham reduced growth of E. coli O157:H7 for 4 weeks. On treated ham the growth of S. typhimurium was increased from week 3. No difference was observed between control and treatment samples with other organisms.