Assessment of anti-biofilm activity and bifidogenic growth stimulator (BGS) effect of lyophilized exopolysaccharides (l-EPSs) from Lactobacilli strains

In the present study, a total of used six lactic acid bacterial strains that isolated from children’s feces and milk products. This study investigated the exopolysacch aride production of these strains. In the culture broth, highest exopolysaccharide production capacity was detected in GD-11, LB-69, and B-3 strains. Monosaccharide compositions of the exopolysaccharides that were purified and lyophilized in the culture medium were investigated and dominant monomer was found to be mannose in all strains. The biofilm-forming capacity of 13 pathogen bacteria was determined and four strains with the highest biofilm-forming capacity were selected as test bacteria for anti-biofilm studies. Different concentrations of l-exopolysaccharides of three lactic acid bacteria were tested for anti-biofilm effect. Highest anti-biofilm effects were exhibited by the L. fermentum LB-69 (90%) against B. cereus RSKK 863. In the statistical analyses performed using the available data, a strong correlation was found between the amount of mannose present in l-exopolysaccharides and the anti-biofilm activity exhibited by l-exopolysaccharides (p < 0.01). In this study, it was concluded that the l-exopolysaccharides of the L. fermentum LB-69 strain could be used as efficient agents with both bifidogenic growth stimulator effect and anti-biofilm effect. If these agents are taken via oral route, it would be possible to provide beneficial effects for the host by increasing the development of bifidobacteria present in the gastrointestinal tract as well as to prevent and treat the diseases by hindering the biofilm formation by harmful bacteria.     

Enzymatic activity of lactic acid bacteria (with antimicrobial properties) isolated from a traditional Spanish cheese

Twenty-four strains of lactic acid bacteria (LAB) isolated from a traditional Spanish cheese (Genestoso cheese) were evaluated for their enzymatic activities (acidifying and proteolytic abilities and carboxypeptidase, aminopeptidase, dipeptidase, caseinolytic and esterase activities), in order to select indigenous strains of technical interest for the manufacture of cheese. These strains were selected on the basis of their antimicrobial activity relative to five reference strains and were identified as Lactococcus lactis subsp. lactis (thirteen strains), Leuconostoc mesenteroides (two strains), Leuconostoc pseudomesenteroides (one strain), Lactobacillus paracasei (two strains), Lactobacillus plantarum (one strain) and Enterococcus faecalis (five strains).     

从蒸馏废液中制造高香气米烧酒(2)

0　前言使用多拷贝糖化酶和耐酸性α -淀粉酶基因的工程菌ＴＲ12菌株 ,烧酒制造中的蒸馏废液可不经排放重复地被利用而制得优质米烧酒 ,但使用常规白曲酶菌 (ＡｓｐｅｒｇｉｌｌｕｓＫａｗａｃｈｉｉ)却未见到同样效果。在蒸馏废液中补充与使用工程菌ＴＲ12菌株时相当量的糖化酶制剂 ,也未能获得更好的收率和取得更好品质的烧酒。在通过对米曲、蒸馏废液以及多种市售酶的比较分析后 ,选取了低酸性蛋白酶含量的一组糖化酶制剂作为添加物 ,进行了实验室规模米烧酒发酵的废液重复利用试验。结果发现 ,蒸馏废液经两次的循环利用 ,在半固体状态的…     

Potential of a lactic acid bacterial starter culture with gamma-aminobutyric acid (GABA) activity for production of fermented sausage

The ability of lactic acid bacterial starter cultures to produce gamma-aminobutyric acid (GABA) during sausage fermentation was studied. Among 305 strains of lactic acid bacteria isolated from kimchi samples, 11 strains were selected as starter candidates based on the following criteria: growth speed, pH lowering ability, and biogenic amine productivity including GABA-producing activity. During in vitro tests, the Y8 (Lactobacillus brevis), O52, and KA20 strains produced 39.00 ± 1.36, 49.73 ± 3.80, and 64.59 ± 0.61 mg/kg of GABA, respectively. Interestingly, although isolate Y8 showed low productivity in vitro, the GABA content it produced during in situ tests (61.30 ± 2.61 mg/kg) was similar to that produced by isolate PM3 (L. brevis) used as positive control (69.64 ± 2.20 mg/kg). Therefore, isolate Y8 was selected as the best functional starter culture for the production of fermented sausage because it exhibited rapid growth, safety, and abundant GABA productivity.     

Antimicrobial activity of neutralized extracellular culture filtrates of lactic acid bacteria isolated from a cultured Indian milk product (‘dahi’)

Neutralized extracellular culture filtrate obtained from isolates of Lactobacillus acidophilus, Lactobacillus delbruecki ssp. bulgaricus, Lactobacillus salivarius and Lactococcus lactis ssp. lactis from ‘dahi’ showed weal to moderate inhibition of Staphylococcus aureus, Bacillus cereus, Escherichia coli, Bacillus brevis, Bacillus circulans, Bacillus coagulans, Bacillus laterosporus, Bacillus subtilis and Pseudomonas aeruginosa when tested by the diffusion agar well assay method. The effective minimum quantity of lactic culture filtrates required to obtain complete inhibition of an inoculum of 10³ cfu/ml of the bacteria tested was between 20 and 26% (vol/vol), as determined by the agar incorporation method. Neutralized extracellular culture filtrate of these lactic cultures added at a level of 10% in sterile, 10% reconstituted non-fat dry milk was able to either suppress or retard growth of selected bacterial cultures when incubated at 37°C for 24 h. This study indicated the antimicrobial activity of dahi and the potential of using neutralized extracellular culture filtrate of lactic acid bacteria in the biopreservation of foods.     

Production and functional evaluation of a protein concentrate from giant squid (Dosidicus gigas) by acid dissolution and isoelectric precipitation

A protein concentrate from giant squid (Dosidicus gigas) was produced under acidic conditions and its functional–technological capability evaluated in terms of its gel-forming ability, water holding capacity and colour attributes. Technological functionality of the concentrate was compared with that of squid muscle and a neutral concentrate. Protein–protein aggregates insoluble at high ionic strength (I = 0.5 M), were detected in the acidic concentrate as result of processing with no preclusion of its gel-forming ability during the sol-to-gel thermal transition. Even though washing under acidic condition promoted autolysis of the myosin heavy chain, the acidic concentrate displayed an outstanding ability to gel giving samples with a gel strength of 455 and 1160 g cm at 75% and 90% compression respectively, and an AA folding test grade indicative of high gel strength, elasticity, and cohesiveness. The process proved to be a good alternative for obtaining a functional protein concentrate from giant squid muscle.     

Assessment of a new lateral flow immunochromatographic (LFIC) assay for the okadaic acid group of toxins using naturally contaminated bivalve shellfish from the Portuguese coast

A new rapid assay for the okadaic acid group of toxins, based on lateral flow immunochromatographic (LFIC) test strips developed by Jellett Rapid Testing Ltd., was assessed on naturally contaminated bivalves from the Portuguese coast. One prototype was evaluated using samples harvested during 2005, extracted with 80% methanol, followed by dilution with the running buffer of a methanolic extract after alkaline hydrolysis for esters. The second prototype was assessed using samples harvested during 2006, extracted with 100% methanol and, after alkaline hydrolysis, the methanol was evaporated by a nitrogen stream followed by re-suspension with the running buffer. The first prototype failed to detect 20% of samples that were positive by LC–MS in the range 160–480 µg kg^?1, and were classified as negative or trace level by LFIC. The presence of methanol in the extracts made correct detection of toxins more difficult. The second prototype classified as positive all samples above 160 µg kg^?1, as confirmed by LC-MS. However, in the second prototype, matrix effects were a major drawback and led to 45% false positives, particularly for mussels, due to compounds in shellfish extracts interfering with the antibodies and reducing the test line intensity. Extraction with a higher percentage of methanol was thought responsible for these matrix effects. Regarding sample migration, both prototypes needed one hour before reading. In an attempt to speed-up sample preparation, a direct digestion of bivalve tissues with sodium hydroxide was evaluated. Low recoveries for esters were found by LC–MS with this hydrolysis technique compared to conventional hydrolysis of methanolic extracts. While prototype A was not sensitive enough, prototype B had too many false positives to be of use to the shellfish industry or in a monitoring program.     

Influence of light exposure, ethanol and copper(II) on the formation of a precursor for xanthylium cations from tartaric acid

The production of xanthylium cation pigments was greatly increased when an aged, tartaric acid buffered, 12% (v/v) aqueous ethanol solution was used in a model white wine system. This suggested the formation of a precursor to the pigments during the ageing of the tartaric acid solution. On examining factors responsible for the generation of tartaric acid oxidation products in wine-like solutions it was observed that on exposure of samples to sunlight, glyoxylic acid, a known precursor to xanthylium cations, was produced. The production of glyoxylic acid was achieved in both the absence and presence of ethanol and copper(II). Hydrogen peroxide was also detected in these solutions. The results were consistent with the presence of glyoxylic acid in the aged tartaric acid buffered, 12% (v/v) aqueous ethanol solution that had frequent aeration and periodic exposure to sunlight throughout its storage. Studies on the role of hydrogen peroxide in the production of glyoxylic acid were also investigated. On the addition of hydrogen peroxide to tartaric acid solutions, with heating at 45°C in darkness, glyoxylic acid was only determined in solutions without ethanol.     

Validation of a tertiary model for predicting variation of Salmonella typhimurium DT104 (ATCC 700408) growth from a low initial density on ground chicken breast meat with a competitive microflora

Growth of a multiple antibiotic-resistant strain (ATCC 700408) of Salmonella Typhimurium definitive phage type 104 (DT104) from a low initial density (10(0.6) most probable number [MPN] or CFU/g) on ground chicken breast meat with a competitive microflora was investigated and modeled as a function of time and temperature (10 to 40 degrees C). MPN and viable counts (CFU) on a selective medium with four antibiotics enumerated the pathogen. Data from five replicate challenge studies per temperature were combined and fit to a primary model to determine maximum specific growth rate (micro), maximum population density (Nmax), and the 95% prediction interval (PI). Nonlinear regression was used to obtain secondary models as a function of temperature for micro, Nmax, and PI, which ranged from 0.04 to 0.4 h(-1), 1.6 to 9.4 log MPN or CFU/g, and 1.4 to 2.4 log MPN or CFU/g, respectively. Secondary models were combined with the primary model to create a tertiary model for predicting variation (95% PI) of pathogen growth among batches of ground chicken breast meat with a competitive microflora. The criterion for acceptable model performance was that 90% of observed MPN or CFU data had to be in the 95% PI predicted by the tertiary model. For data (n=344) used in model development, 93% of observed MPN and CFU data were in the 95% PI predicted by the tertiary model, whereas for data (n=236) not used in model development but collected using the same methods, 94% of observed MPN and CFU data were in the 95% PI predicted by the tertiary model. Thus, the tertiary model was successfully verified against dependent data and validated against independent data for predicting variation of Salmonella Typhimurium DT104 growth among batches of ground chicken breast meat with a competitive microflora and from a low initial density.     

A simple method to recover Norovirus from fresh produce with large sample size by using histo-blood group antigen-conjugated to magnetic beads in a recirculating affinity magnetic separation system (RCAMS)

Human norovirus (NoV) outbreaks are major food safety concerns. The virus has to be concentrated from food samples in order to be detected. PEG precipitation is the most common method to recover the virus. Recently, histo-blood group antigens (HBGA) have been recognized as receptors for human NoV, and have been utilized as an alternative method to concentrate human NoV for samples up to 40 mL in volume. However, to wash off the virus from contaminated fresh food samples, at least 250 mL of wash volume is required. Recirculating affinity magnetic separation system (RCAMS) has been tried by others to concentrate human NoV from large-volume samples and failed to yield consistent results with the standard procedure of 30 min of recirculation at the default flow rate. Our work here demonstrates that proper recirculation time and flow rate are key factors for success in using the RCAMS. The bead recovery rate was increased from 28% to 47%, 67% and 90% when recirculation times were extended from 30 min to 60 min, 120 min and 180 min, respectively. The kinetics study suggests that at least 120 min recirculation is required to obtain a good recovery of NoV. In addition, different binding and elution conditions were compared for releasing NoV from inoculated lettuce. Phosphate-buffered saline (PBS) and water results in similar efficacy for virus release, but the released virus does not bind to RCAMS effectively unless pH was adjusted to acidic. Either citrate-buffered saline (CBS) wash, or water wash followed by CBS adjustment, resulted in an enhanced recovery of virus. We also demonstrated that the standard curve generated from viral RNA extracted from serially-diluted virus samples is more accurate for quantitative analysis than standard curves generated from serially-diluted plasmid DNA or transcribed-RNA templates, both of which tend to overestimate the concentration power. The efficacy of recovery of NoV from produce using RCAMS was directly compared with that of the PEG method in NoV inoculated lettuce. 40, 4, 0.4, and 0.04 RTU can be detected by both methods. At 0.004 RTU, NoV was detectable in all three samples concentrated by the RCAMS method, while none could be detected by the PEG precipitation method. RCAMS is a simple and rapid method that is more sensitive than conventional methods for recovery of NoV from food samples with a large sample size. In addition, the RTU value detected through RCAMS-processed samples is more biologically relevant.