Evaluating the Probiotic Potential of Lactic Acid Bacteria Implicated in Natural Fermentation of Table Olives, cv. Cobrançosa.

The probiotic features of Lactiplantibacillus (L.) pentosus and L. paraplantarum strains, endogenous in Cobrançosa table olives from northeast Portugal, were assessed in terms of functional properties and health benefits. Fourteen lactic acid bacteria strains were compared with Lacticaseibacillus casei from a commercial brand of probiotic yoghurt and L. pentosus B281 from Greek probiotic table olives, in attempts to select strains with higher probiotic performances than those references. For functional properties, the i53 and i106 strains, respectively, exhibited: 22.2 ± 2.2% and 23.0 ± 2.2% for Caco-2 cell adhesion capacity; 21.6 ± 7.8% and 21.5 ± 1.4% for hydrophobicity; 93.0 ± 3.0% and 88.5 ± 4.5% for autoaggregation ability by 24 h of incubation; and ability to co-aggregate with selected pathogens-from 29 to 40% to Gram+ (e.g., Staphylococcus aureus ATCC 25923 and Enterococcus faecalis ATCC 29212); and from 16 to 44% for Gram- (e.g., Escherichia coli ATCC 25922 and Salmonella enteritidis ATCC 25928). The strains proved to be resistant (i.e., halo zone ≤14 mm) to some antibiotics (e.g., vancomycin, ofloxacin, and streptomycin), but susceptible (i.e., halo zone ≥ 20 mm) to others (e.g., ampicillin and cephalothin). The strains exhibited health-beneficial enzymatic activity (such as acid phosphatase and naphthol-AS-BI-phosphohydrolase), but not health-harmful enzymatic activity (such as ß-glucuronidase and N-acetyl-ß-glucosaminidase). Additionally, the antioxidant activity and cholesterol assimilation features, respectively, of the strains were 19.6 ± 2.8% and 77.5 ± 0.5% for i53, and 19.6 ± 1.8% and 72.2 ± 0.9% for i106. This study indicated that the addition of L. pentosus strains i53 and/or i106 to Cobrançosa table olives is likely to enhance the added value of the final product, in view of the associated potential benefits upon human health.

Study of Lactic Acid Bacteria Biodiversity in Fermented Cobrançosa Table Olives to Determine Their Probiotic Potential.

Current market trends point at increasing demand for functional foods, namely those carrying probiotics. In the case of table olives, presence of probiotics would convey a competitive advantage to Mediterranean-based diets, already established for their cultural heritage and gastronomic character. This work assessed the safety and resistance to gastrointestinal digestion of 19 native LAB strains from Cobrançosa table olives. Strains were identified via molecular sequencing (4 fingerprints/10 strains for Lactiplantibacillus pentosus, and 2 fingerprints/9 strains for L. paraplantarum), and exposed to simulated gastrointestinal fluids, as per the INFOGEST in vitro protocol with modifications. None of those strains proved dangerous for human consumption. Survivability to the gastrointestinal resistance test ranged from 29% to 70%, with strain-dependent variability. L. paraplantarum i18, i27, and i102, and L. pentosus i10 and i11 exhibited statistically lower survival rates (29−35%) than probiotic the Greek table olive reference strain L. pentosus B281 (53%). Among the other strains, L. paraplantarum i101 and L. pentosus i53 and i106 showed the highest survival rates but were not significantly different from the strain of Lacticaseibacillus casei isolated from commercial probiotic yoghurt (65−70%). In vitro results proved that strains retrieved from fermenting cultivar Cobrançosa possess the potential to be claimed as probiotics­thus deserving further attention toward the development of a specific starter culture.

Insights on sulfamethoxazole bio-transformation by environmental Proteobacteria isolates.

Although sulfonamide residues are frequently reported as freshwaters contaminants, information on the ability of native bacteria to modify these synthetic antibiotics is scarce. Our purpose was to investigate the potential of bacteria from different aquatic environments to cleave or transform sulfamethoxazole (SMX) and infer on their ability to reduce the toxicity of this antibiotic. From a collection of about 100 Proteobacteria, 47 strains previously isolated from drinking water, surface water, and wastewater grew in the presence of 200 µMSMX, and were further studied. Out of these, 14 strains, mostly from mineral drinking water, transformed SMX into equimolar amounts of the lesser toxic derivative N4-acetyl-sulfamethoxazole. The highest percentage of SMX transformation was recorded for two strains affiliated to Pseudomonas mandelii. For P. mandelii McBPA4 higher SMX transformation rate and extent were observed in fed-batch (∼8 µMSMX/h, 81%) than in batch conditions (∼5 µMSMX/h, 25%), but similar specific transformation rates were found in both cultivation modes (∼20 µmolSMX/gcell dry weight/h), indicating the dependence of the process on the microbial load. These results evidence that the capacity to transform synthetic antibiotics may be common among bacteria and highlight the potential of environmental bacteria in attenuating the potential adverse effects of pollution with sulfonamides.

Biodegradation of sulfamethoxazole and other sulfonamides by Achromobacter denitrificans PR1.

This study aimed to isolate and characterize a microbial culture able to degrade sulfonamides. Sulfamethoxazole (SMX)-degrading microorganisms were enriched from activated sludge and wastewater. The resultant mixed culture was composed of four bacterial strains, out of which only Achromobacter denitrificans PR1 could degrade SMX. This sulfonamide was used as sole source of carbon, nitrogen and energy with stoichiometric accumulation of 3-amino-5-methylisoxazole. Strain PR1 was able to remove SMX at a rate of 73.6 ± 9.6 µmol SMX/gcell dryweighth. This rate more than doubled when a supplement of amino acids or the other members of the mixed culture were added. Besides SMX, strain PR1 was able to degrade other sulfonamides with anti-microbial activity. Other environmental Achromobacter spp. could not degrade SMX, suggesting that this property is not broadly distributed in members of this genus. Further studies are needed to shed additional light on the genetics and enzymology of this process.