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.

Effect of Different Lactic Acid Bacteria on Phytic Acid Content and Quality of whole Wheat Toast Bread

Nowadays, consumption of whole flours and flours with high extraction rate is recommended, because of their high amount of fibers, vitamins and minerals. Despite nutritional benefits of whole flour, concentration of some undesirable components such as phytic acid is higher than white flour. In this study, effect of several lactic acid bacteria sourdough on Toast bread is investigated. Sourdough from lactic acid bacteria (Lb. plantarum, Lb. reuteri) with different dough yield (250 and 300) is made and incubated at 30°C for 20 hour, then added to dough in the ratio of 10, 20 and 30% replacement. Breads that supplemented with Lb. plantarum sourdough had lower phytic acid. Higher replacement of sourdough and higher DY cause higher decrease in phytic acid content. Sourdough from Lb. plantarum, DY = 300 and 30% replacement cause the highest decrease in phytic acid content (49.63 mg/100g). As indicated by panelists, Lb. reuteri sourdough can present the greatest effect on overall quality score of the breads. DY reduction cause a decrease in bread quality score. Sensory score of Toast bread is 81.71 in the samples that treated with Lb. reuteri sourdough with DY = 250 and 20% replacement.