Development of enterococci and production of tyramine during the manufacture and ripening of Cheddar cheese

peer-reviewedThe effect of six strains of enterococci (three strains of Enterococcus faecalis, and one strain each of Ec. faecium, Ec. durans and Ec. casseliflavus) on flavour development and tyramine production in Cheddar cheese during manufacture and ripening was studied in two trials. No strain produced gelatinase or haemolysin and all of them grew well during manufacture reaching 107 colony forming units (cfu)/g in 6 h, after which they remained more or less constant during at least 48 weeks of ripening. There was no relationship between tyramine production in a broth containing tyrosine and tyramine production in the cheese. All strains, except Ec. casseliflavus, produced tyramine in the cheese, with the greatest concentration (162 mg/kg) being produced by Ec. durans after 9 months ripening at 8 ºC. There was no statistically significant difference (P > 0.05) between the flavour of the control cheese and any cheese containing an enterococcus. Nevertheless, cheese made with Ec. faecium E-24 received the best score in each trial at both time points. No off-flavours were found. Regarding proteolysis, only Ec. faecalis E-140 showed significant (P < 0.05) increases in both phosphotungstic acid and pH 4.6 soluble N. It is concluded that enterococci have little effect on the flavour of Cheddar cheese.This project was partly financed under FAIR Contract CT97-3078 from the E

Three New Escherichia coli Phages from the Human Gut Show Promising Potential for Phage Therapy

peer-reviewedWith the emergence of multi-drug resistant bacteria the use of bacteriophages (phages) is gaining renewed interest as promising anti-microbial agents. The aim of this study was to isolate and characterize phages from human fecal samples. Three new coliphages, ɸAPCEc01, ɸAPCEc02 and ɸAPCEc03, were isolated. Their phenotypic and genomic characteristics, and lytic activity against biofilm, and in combination with ciprofloxacin, were investigated. All three phages reduced the growth of E. coli strain DPC6051 at multiplicity of infection (MOI) between 10−3 and 105. A cocktail of all three phages completely inhibited the growth of E. coli. The phage cocktail also reduced biofilm formation and prevented the emergence of phage-resistant mutants which occurred with single phage. When combined with ciprofloxacin, phage alone or in cocktail inhibited the growth of E. coli and prevented the emergence of resistant mutants. These three new phages are promising biocontrol agents for E. coli infections

Expression of the malolactic enzyme gene (mle) from Lactobacillus plantarum under winemaking conditions

Malolactic fermentation (MLF) plays an important role in the production of wine, especially red wines, resulting in microbial stability, deacidification, as well as contributing to the aroma profile. MLF can be influenced by a number of factors. In this study, the influence of pH and ethanol on expression of the structural malolactic enzyme gene (mle) from Lactobacillus plantarum was investigated in a synthetic wine media, as well as in wine using quantitative PCR. Expression of mle was shown to be inducible by the presence of malic acid, with increased expression in the middle of MLF. Expression of mle was also shown to be increased at low pH values and decreased in the presence of ethanol. This indicates the role of MLF in acid tolerance and the negative impact of ethanol on the completion of MLF. The results therefore provide further evidence that L. plantarum should be applied as co-inoculation for MLF where alcohol will initially not have a negative impact on the malic acid degradation. © 2011 Springer Science+Business Media, LLC.Articl

Development of a quantitative PCR for detection of Lactobacillus plantarum starters during wine malolactic fermentation

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Characterisation and selection of probiotic lactobacilli for a preliminary minipig feeding trial and their effect on serum cholesterol levels, faeces pH and faeces moisture content

Three out of 297 Lactobacillus strains isolated from pig faeces were selected for a feeding trial on account of their high bile-salt hydrolase (BSH) activity, bile-salt resistance, low pH tolerance and the production of antimicrobial substances. Two strains were identified as Lactobacillus johnsonii and one as Lactobacillus reuteri by DNA-DNA hybridisation. L. johnsoniii BFE 1061 produced a bacteriocin active against a range of lactic acid bacteria (LAB) and nonrelated bacteria including Clostridium perfringens. Six minipigs were maintained on a high-fat, high-cholesterol ('Western Style') diet for 17 weeks after which the diet was supplemented with the 'probiotic mixture' containing the above mentioned three Lactobacillus strains at 2 x 1012 CFU per pig per day for five weeks. The mixture was given as a resuspended lyophilisate. During a two week follow-up period the minipigs received only the 'Western-style' diet without probiotic supplementation. A lowering effect on serum cholesterol levels was indicated after three weeks probiotic feeding, concomitant with an increase in the moisture content of the faeces and Lactobacillus cell numbers. Triglycerides, pH and number of lactic acid bacteria in faeces were not significantly influenced by probiotic supplementation.Articl