Abstract Four bacteriocin producing lactic acid bacteria isolated from vegetables were identified as Lactococcus lactis strains on the basis of physiological and biochemical characteristics, carbohydrate fermentation patterns and analysis of total soluble protein pattern by SDS PAGE. The bacteriocins had a wide spectrum of activity as antagonism was detected not only towards a variety of lactic acid bacteria, but also to Staphylococcus aureus and Listeria monocytogenes . These bacteriocins were resistant to heating at 121 °C for 15 minutes and showed highest activity at low pH (<5.0). They were inactivated by the proteolytic enzymes α‐chymotrypsin and proteinase K, but not by lipase, α‐amylase, catalase or lysozyme. These bacteriocinogenic Lactococcus strains were all immune to the bacteriocins produced as well as to commerical nisin. Bacteriocin producer culture supernatants showed a high degree (70 or 100%) of cross‐reactivity in the nisin ELISA, suggesting similarity of the produced bacteriocins to nisin. The potential application of bacteriocin producing lactococci of vegetable origin for safety assurance of vegetable foods and controlling vegetable fermentations is discussed.
To test the effects of C : N : P ratio modification of a well-known nutrient medium formulation, the Endo formulation on biofilm formation by Enterobacter cloacae Ecl and Citrobacter freundii Cf1 in both single-species and binary species biofilms.The C : N : P atom : atom ratio of a well-known nutrient medium formulation, the Endo formulation, that has been applied in fermentative biohydrogen studies, was modified to include two different C concentrations, one containing 17.65 g l(-1) and the other 8.84 g l(-1) sucrose, each containing four different C : N : P ratios, two at higher C : N : P ratios (334 : 84 : 16.8 and 334 : 84 : 3) and two at lower C : N : P ratios (334 : 28 : 5.6 and 334 : 28 : 1). Attached cells were enumerated after dislodging the biofilms that had formed on granular activated carbon (GAC). The modified medium containing 17.65 g l(-1) sucrose and having a C : N : P ratio of 334 : 28 : 5.6 resulted in significantly (P < 0.05) higher counts of attached cells for both single-species biofilms at 7.73 log(10) CFU g(-1) GAC and 9.3 log(10)CFU g(-1) GAC for Ent. cloacae Ecl and Cit. freundii Cf1, respectively, and binary species biofilms at 8.2 log(10) CFU g(-1) GAC and 6.34 log(10) CFU g(-1) GAC for Ent. cloacae Ecl and Cit. freundii Cf1, respectively. Scanning electron micrographs showed qualitative evidence that the 334 : 28 : 5.6 ratio encouraged more complex and extensive biofilm growth for both single-species and binary species biofilms.The differences in the attachment numbers between the different ratios were found not to be a result of the individual actions of the bacterial isolates involved but rather because of the effects of the various C : N : P ratios. The 334 : 28 : 5.6 ratio showed significantly (P < 0.05) higher counts of attached cells for both single-species and binary species biofilms.This study indicates that C : N : P ratios should be a key consideration with regard to maximizing biofilm formation in shake flask and fluidized bed bioreactor studies as well as understanding fundamental factors affecting biofilm growth in natural environments.
Daily exposure to 100 p.p.m. chlorine dioxide of single species and binary biofilms of dairy-associated Bacillus cereus DL5 and Pseudomonas fluorescens M2, attached to stainless steel surfaces in a laboratory flow system, was studied.Surfaces were sampled daily before and after sanitizer treatment and cells and spores dislodged and enumerated by standard methods. Duplicate surfaces were prepared for confocal scanning laser microscopy (CSLM) and scanning electron microscopy. Higher counts of Ps. fluorescens M2 were obtained in single species biofilms, microcolonies stained green (viable) in CSLM images and were closely packed on attachment surfaces. By contrast, higher counts of B. cereus DL5 were obtained in binary biofilms, microcolonies stained green in CSLM images, but were more spread out. Lower spore counts were obtained for B. cereus DL5 in binary biofilms. The survival of Ps. fluorescens M2 cells after exposure to chlorine dioxide was apparently enhanced by the presence of B. cereus DL5 in binary biofilms. By contrast, B. cereus DL5 showed increased susceptibility to sanitizer treatment in the presence of Ps. fluorescens M2.Co-cultured bacteria in biofilms influence each other with respect to attachment capabilities and sanitizer resistance/susceptibility.Binary biofilms endemic in food-processing industries can survive sanitation regimes and may represent reservoirs of product contamination leading to subsequent spoilage and/or food safety risks.
Abstract This study assessed the in vitro responses of Bacillus (B.) strains isolated from ropey bread to natural antimicrobials under optimum growth conditions. The responses of six Bacillus strains [ B. subtilis (2), B. licheniformis (2) and B. pumilus (2)] to acetic acid (AA), lactic acid (LA), calcium lactate (CL) and a lactate‐containing cocktail (LCC), singly and in combinations were determined and compared to calcium propionate (CP). Isolates were each inoculated into flasks containing Nutrient Broth (NB) and the respective antimicrobial treatments and pHs were left unadjusted. A duplicate set of flasks, also containing NB and the respective antimicrobials, but adjusted to pH's corresponding to those of baked brown bread containing the same antimicrobials was also inoculated. Growth curves were obtained spectrophotometrically and used to estimate lag times. The organic acids used in this study [0.1% (v/v) AA and 0.25% (v/v) LA] singly and in combination with each other and with CL, CP or LCC, completely inhibited the growth of all six Bacillus strains, but only at non‐adjusted pHs. The efficacies of LA, AA and CL notably decreased when the pH of the test media containing the respective preservatives was adjusted to the corresponding in situ (bread) values. However, the natural antimicrobials were still as effective as CP in retarding growth of the six Bacillus strains at the in situ (bread) pH values.
Rope and mould spoilage decrease the shelf life of bread and may result in substantial monetary losses to producers. Five test bakes were carried out to determine the efficacy of sodium diacetate or vinegar in combination with calcium propionate, in inhibiting rope and mould spoilage of brown bread. Their effect on yeast activity was also evaluated. Baked loaves were stored at 30 degrees C and analysed for 6 days post-baking by duplicate plate counting, viable staining using 0.5% tetrazolium salt solution, mould-free shelf life determination and pH measurement. Changes in bacterial counts over time were used to rank treatments statistically. Vinegar combined with calcium propionate at 0.10% each achieved the longest rope-free shelf life, while 0.30% calcium propionate resulted in the longest mould-free shelf life. A combination of sodium diacetate and calcium propionate at 0.10% each showed the lowest reduction in yeast activity.
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