Exploratory Tests of Crude Bacteriocins from Autochthonous Lactic Acid Bacteria against Food-Borne Pathogens and Spoilage Bacteria

The aim of the present work was to test in vitro inhibition of food pathogens and spoilage bacteria by crude bacteriocins from autochthonous lactic acid bacteria. Thirty autochthonous lactic acid bacteria isolated previously, belonging to the genera: Lactobacillus, Carnobacterium, Lactococcus, Vagococcus, Streptococcus, and Pediococcus, have been screened by an agar spot test and a well diffusion assay against Gram-positive and Gram-negative harmful bacteria: Bacillus cereus, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 8739, Salmonella typhimurium ATCC 14028, Staphylococcus aureus ATCC 6538, and Pseudomonas aeruginosa under conditions means to reduce lactic acid and hydrogen peroxide effect to select bacteria with high bacteriocinogenic potential. Furthermore, crude bacteriocins semiquantification and heat sensitivity to different temperatures (80, 95, 110°C, and 121°C) were performed. Another exploratory test concerning the response of St. aureus ATCC 6538 to the presence of crude bacteriocins was realized. It has been observed by the agar spot test that fifteen candidates were active toward Gram-positive targets strains. The secondary screening demonstrated an antagonistic activity oriented only against St. aureus ATCC 6538, leading to the selection of five isolates: Lm14, Lm21, Lm23, Lm24, and Lm25 with a larger inhibition zone compared to the others. The ANOVA statistical analysis reveals a small variation of repeatability: Lm21: 0.56%, Lm23: 0%, Lm25: 1.67%, Lm14: 1.88%, Lm24: 2.14%. Conversely, slight variation was reported in terms of inhibition diameters: 9.58± 0.40, 9.83± 0.46 and 10.16± 0.24 8.5 ± 0.40 10 mm for, Lm21, Lm23, Lm25, Lm14and Lm24, indicating that the observed potential showed a heterogeneous distribution (BMS = 0.383, WMS = 0.117). The repeatability coefficient calculated displayed 7.35%. As for the bacteriocins semiquantification, the five samples exhibited production amounts about 4.16 for Lm21, Lm23, Lm25 and 2.08 AU/ml for Lm14, Lm24. Concerning the sensitivity the crude bacteriocins were fully insensitive to heat inactivation, until 121°C, they preserved the same inhibition diameter. As to, kinetic of growth , the µmax showed reductions in pathogens load for Lm21, Lm23, Lm25, Lm14, Lm24 of about 42.92%, 84.12%, 88.55%, 54.95%, 29.97% in the second trails. Inversely, this pathogen growth after five hours displayed differences of 79.45%, 12.64%, 11.82%, 87.88%, 85.66% in the second trails, compared to the control. This study showed potential inhibition to the growth of this food pathogen, suggesting the possibility to improve the hygienic food quality.

The Effect of Sodium Chloride and pH on the Antimicrobial Effectiveness of Essential Oils Against Pathogenic and Food Spoilage Bacteria:Implications in Food Safety

The purpose of this study was to elucidate the factors affecting antimicrobial effectiveness of essential oils against food spoilage and pathogenic bacteria. The minimum inhibition concentrations (MIC) of the essential oils, were determined by turbidimetric technique using Biocreen C, analyzer. The effects of pH ranging from 7.3 to 5.5 in absence and presence of essential oils and/or NaCl on the lag time and mean generation time of the bacteria at 370C, were carried out and results were determined showed that, combination of low pH and essential oil at 370C had additive effects against the test micro-organisms. The combination of 1.2 % (w/v) of NaCl and clove essential oil at 0.0325% (v/v) was effective against E. coli. The use of concentrations less than MIC in combination with low pH and or NaCl has the potential of being used as an alternative to “traditional food preservatives".

Inhibitory Effect of Lactic Acid and Nisin on Bacterial Spoilage of Chilled Shrimp

Lactic acid alone and its combined application with nisin were evaluated for reducing population of naturally occurring microorganisms on chilled shrimp. Fresh shrimps were dipped in 0, 1.0% and 2.0% (v/v) lactic acid alone and their combined application with 0.04 (g/L/kg) nisin solution for 10 min. Total plate counts of aerobic bacteria (TPCs), Psychrotrophic counts, population of Pseudomonas spp., H2S producing bacteria and Lactic acid bacteria (LAB) on shrimps were determined during storage at 4 °C. The results indicated that total plate counts were 2.91 and 2.63 log CFU/g higher on untreated shrimps after 7 and 14 days of storage, respectively, than on shrimps treated with 2.0% lactic acid combined with 0.04 (g/L/kg) nisin. Both concentrations of lactic acid indicated significant reduction on Pseudomonas counts during storage, while 2.0% lactic acid combined with nisin indicated the highest reduction. In addition, H2S producing bacteria were more sensitive to high concentration of lactic acid combined with nisin during storage.