There has been continued interest in bacteriocins research from an applied perspective as bacteriocins have potential to be used as natural preservative. Four bacteriocinogenic lactic acid bacteria (LAB) strains of Lactobacillus curvatus (Arla-10), Enterococcus faecium (JFR-1), Lactobacillus paracasei subsp. paracasei (JFR-5) and Streptococcus thermophilus (TSB-8) were previously isolated and identified in our lab. The objective of this study was to determine the optimal growth conditions for both LAB growth and bacteriocins production. In this study, various growth conditions including culture media (MRS and BHI), initial pH of culture media (4.5, 5.5, 6.2, 7.4 and 8.5), and incubation temperatures (20, 37 and 44 °C) were investigated for LAB growth measured as optical density (OD), bacteriocin activity determined as arbitrary unit and viability of LAB expressed as log CFU ml−1. Growth curves of the bacteriocinogenic LAB were generated using a Bioscreen C. Our results indicated that Arla-10, JFR-1, and JFR-5 strains grew well on both MRS and BHI media at growth temperature tested whereas TSB-8 strain, unable to grow at 20 °C. LAB growth was significantly affected by the initial pH of culture media (p < 0.001) and the optimal pH was found ranging from 6.2 to 8.5. Bacteriocin activity was significantly different in MRS versus BHI (p < 0.001), and the optimal condition for LAB to produce bacteriocins was determined in MRS broth, pH 6.2 at 37 °C. This study provides useful information on potential application of bacteriocinogenic LAB in food fermentation processes.

The biopreservation of foods using bacteriocinogenic lactic acid bacteria (LAB) isolated directly from foods is an innovative approach. The objectives of this study were to isolate and identify bacteriocinogenic LAB from various cheeses and yogurts and evaluate their antimicrobial effects on selected spoilage and pathogenic microorganisms in vitro as well as on a food commodity. LAB were isolated using MRS and M17 media. The agar diffusion bioassay was used to screen for bacteriocin or bacteriocin-like substances (BLS) producing LAB using Lactobacillus sakei and Listeria innocua as indicator organisms. Out of 138 LAB isolates, 28 were found to inhibit these bacteria and were identified as strains of Enterococcus faecium, Streptococcus thermophilu s, Lactobacillus casei and Lactobacillus sakei subsp. sakei using 16S rRNA gene sequencing. Eight isolates were tested for antimicrobial activity at 5°C and 20°C against L. innocua, Escherichia coli, Bacillus cereus, Pseudomonas fluorescens, Erwinia carotovora, and Leuconostoc mesenteroides subsp. mesenteroides using the agar diffusion bioassay, and also against Penicillium expansum, Botrytis cinerea and Monilinia frucitcola using the microdilution plate method. The effect of selected LAB strains on L. innocua inoculated onto fresh-cut onions was also investigated. Twenty percent of our isolates produced BLS inhibiting the growth of L. innocua and/or Lact. sakei. Organic acids and/or H₂O₂ produced by LAB and not the BLS had strong antimicrobial effects on all microorganisms tested with the exception of E. coli. Ent. faecium, Strep. thermophilus and Lact. casei effectively inhibited the growth of natural microflora and L. innocua inoculated onto fresh-cut onions. Bacteriocinogenic LAB present in cheeses and yogurts may have potential to be used as biopreservatives in foods.

Canada is a leading producer of wild lowbush blueberries, most of which are mechanically harvested, washed, individually quick frozen (IQF), and bulk packaged. Still, some berries are harvested by more gentle methods and sold as fresh-packed products. These berries do not undergo a wash step, nor are subjected to antimicrobial treatments. The purpose of this study was to conduct a microbiological survey of berries harvested in the province of Nova Scotia to assess their potential for harborage of bacterial foodborne pathogens. A combination of standardized plate count methods and 3M-Petrifilm protocols were used to enumerate total aerobic mesophilic bacteria (APC), yeasts and molds (YMC), coliforms, and generic E. coli, the latter being an indicator of fecal contamination. Overall, APC and YMC levels were 1.2 and 0.5 log greater, respectively, for berries collected early in the harvest season versus those acquired late season and varied significantly (p < 0.05) between farm (location) and harvest practices used. Berries harvested by our team using sanitized hand rakes (SH) had consistently lower APC and YMC levels than those harvested by farm crews. Yet, when gentle harvesting (GH) methods (hand-raking, walk-behind or modified mechanical harvesters) were employed on farms, lower numbers were generally observed compared to berries harvested by traditional tractor-mounted mechanized harvesters (MH). The presence of coliforms (and their levels) was also impacted by the harvest method, with detection rates of ~29%, 73%, and 92% in SH, GH, and MH samples, respectively. Mean counts were < 2.5 log₁₀ CFU/g for both SH and GH berries, but significantly higher (p < 0.05) on MH berries (3.6 log₁₀ CFU/g). Although ~56% of all berry samples collected (n = 350) contained coliforms, only 12 were positive for E. coli, 9 of which were MH samples. Only the latter had numbers > 2 log₁₀ CFU/g, but none tested positive for Shiga toxin-producing serotype O157 (STEC O157) or Salmonella spp. when using internationally recognized selective enrichment and plating methods. ATP luminescence was used to assess the general hygiene of processing lines, whereby “hot spots” for microbial activity were identified, even after cleaning., Standard selective enrichment and plating methods were used for the detection of Listeria monocytogenes on 61 swab samples taken from berry totes or conveyor belts at different times during processing; 4 swabs tested positive for L. monocytogenes. However, the pathogen could not be detected by direct plating on selective agar without prior enrichment; this indicated its numbers were low. The results from this work demonstrated that alternative gentle harvest methods can reduce microbial numbers on wild blueberries. Although the frequency of fecal contamination in berry samples appeared to be low and targeted human pathogens were not detected; this represents a single study conducted over one harvest season. Therefore, it would be prudent for processors to seek effective antimicrobial technologies prior to packaging, while consumers should use caution and thoroughly wash produce before consumption. Where sporadic detection of L. monocytogenes was observed on environmental samples from the processing line, processors must ensure that effective sanitation programs are implemented to avoid potential food safety risks.