Antibacterial and Antiparasitic Activity of Propyl-Propane-Thiosulfinate (PTS) and Propyl-Propane-Thiosulfonate (PTSO) from Allium cepa against Gilthead Sea Bream Pathogens in In Vitro and In Vivo Studies.

The use of phytogenic extracts is considered a sustainable strategy for the prevention of fish diseases, including Alliaceae as a potential option due to their variety of bioactive compounds. In this study, we analyzed the antibacterial and antiparasitic potential of propyl-propane-thiosulfinate (PTS) and propyl-propane-thiosulfonate (PTSO) from onions. The in vitro activity against Pseudomonas anguilliseptica, Tenacibaculum maritimum, and Photobacterium damselae of both compounds was tested. In addition, the viability of Sparicotyle chrysophrii larvae was evaluated. Moreover, a diet that consisted of a blend of PTS/PTSO (ALLIUM) was used. A total of 90 gilthead sea bream juveniles were tested against P. damselae subsp. Piscicida after 12 weeks of dietary administration. Furthermore, 150 fish with a rate of 10-15 parasites/fish were fed for 21 days and the number of gill parasites was recorded. All strains were sensitive to both compounds. PTSO showed the highest inhibitory effect against all target strains, while PTS showed higher effectiveness against S. chrysophrii. Fish from ALLIUM group presented the highest probability of survival, increasing up to 91.1%, whereas in the control group, the probability of survival was 66.7%. The number of parasites in the gilthead sea bream decreased in the ALLIUM group over time. These results suggest the inclusion of PTS and PTSO in feed as a natural strategy to prevent antibacterial and antiparasitic fish diseases.

Phytochemicals Determination, and Antioxidant, Antimicrobial, Anti-Inflammatory and Anticancer Activities of Blackberry Fruits.

A comprehensive characterization of the phytochemicals present in a blackberry fruit extract by HPLC-TOF-MS has been carried out. The main compounds in the extract were ursane-type terpenoids which, along with phenolic compounds, may be responsible for the bioactivity of the extract. In vitro antioxidant capacity was assessed through Folin-Ciocalteu (31.05 ± 4.9 mg GAE/g d.w.), FRAP (637.8 ± 3.2 µmol Fe2+/g d.w.), DPPH (IC50 97.1 ± 2.4 µg d.w./mL) and TEAC (576.6 ± 8.3 µmol TE/g d.w.) assays. Furthermore, the extract exerted remarkable effects on in vitro cellular antioxidant activity in HUVEC cells at a concentration of 5 mg/mL. Antimicrobial activity of the extract was also tested. Most sensible microorganisms were Gram-positive bacteria, such as E. faecalis, B. cereus and Gram-negative E. coli (MBC of 12.5 mg/mL). IC50 values against colon tumoral cells HT-29 (4.9 ± 0.2 mg/mL), T-84 (5.9 ± 0.3 mg/mL) and SW-837 (5.9 ± 0.2 mg/mL) were also obtained. Furthermore, blackberry extract demonstrated anti-inflammatory activity inhibiting the secretion of pro-inflammatory IL-8 cytokines in two cellular models (HT-29 and T-84) in a concentration-dependent manner. These results support that blackberry fruits are an interesting source of bioactive compounds that may be useful in the prevention and treatment of different diseases, mainly related to oxidative stress.

Establishment of a Sonotrode Extraction Method and Evaluation of the Antioxidant, Antimicrobial and Anticancer Potential of an Optimized Vaccinium myrtillus L. Leaves Extract as Functional Ingredient.

Vaccinium myrtillus L. (bilberry) leaves are an important by-product of berry production that may be used as a source of phenolic compounds which have a positive effect on human health. Therefore, an ultrasound-assisted extraction via sonotrode has been used for the first time to recover bioactive compounds from bilberry leaves. The extraction has been optimized using a Box-Behnken design. The influence of ethanol:water ratio (v/v), time of extraction (min) and amplitude (%) were evaluated considering total phenolic content (TPC) and antioxidant capacity (DPPH and FRAP assays) as dependent variables in a response surface methodology (RSM). Optimum values for the independent factors were 30:70 ethanol/water (v/v), 5 min of extraction and 55% amplitude. The empirical values of the independent variables using the optimized conditions were 217.03 ± 4.92 mg GAE/g d.w. (TPC), 271.13 ± 5.84 mg TE/g d.w. (DPPH) and 312.21 ± 9.30 mg TE/g d.w. (FRAP). The validity of the experimental design was confirmed using ANOVA and the optimal extract was characterized using HPLC-MS. A total of 53 compounds were tentatively identified, of which 22 were found in bilberry leaves for the first time. Among them, chlorogenic acid was the most abundant molecule, representing 53% of the total phenolic compounds identified. Additionally, the antimicrobial and anticancer activities of the optimum extract were tested. Gram-positive bacteria demonstrated high sensitivity to bilberry leaves extract in vitro, with MBC values of 6.25 mg/mL for Listeria monocytogenes, Listeria innocua and Enterococcus faecalis, and 0.8 mg/mL for Staphylococcus aureus and Bacillus cereus. Furthermore, bilberry leaves extract exerted in vitro antiproliferative activity against HT-29, T-84 and SW-837 colon tumor cells with IC50 values of 213.2 ± 2.5, 1140.3 ± 5.2 and 936.5 ± 4.6 µg/mL, respectively. Thus, this rapid ultrasound-assisted extraction method has demonstrated to be an efficient technique to obtain bilberry leaves extract with in vitro antioxidant, antimicrobial and anticancer capacities that may be useful for the food industry as natural preservative or even for the production of functional foods or nutraceuticals.

In Vitro Antiparasitic Activity of Propyl-Propane-Thiosulfinate (PTS) and Propyl-Propane-Thiosulfonate (PTSO) from Allium cepa against Eimeria acervulina Sporozoites.

Among the alternatives to control avian coccidiosis, alliaceous extracts stand out due to their functional properties. Despite this, most of the references are focused just on garlic. In this study, we analyze the in vitro effects of propyl-propane thiosulfinate (PTS) and propyl-propane thiosulfonate (PTSO), two organosulfur compounds from onion, on MDBK cells infected with sporozoites of Eimeria acervulina. To this aim, two different experiments were performed. In the first experiment, sporozoites were previously incubated for 1 h at 1, 5 and 10 µg/mL of PTS or PTSO and added to MDBK cells. In the second experiment, MDBK cells were first incubated for 24 h at different concentrations of PTS or PTSO and then infected with E. acervulina sporozoites. Then, 24 h after inoculation, the presence of E. acervulina was quantified by qPCR. MDBK viability was measured at 72 h post-infection. Sporozoites incubated at 10 µg/mL of PTS and PTSO inhibited the capability to penetrate the cells up to 75.2% ± 6.44 and 71.7% ± 6.03, respectively. The incubation of MDBK with each compound resulted in a preventive effect against sporozoite invasion at 1 µg/mL of PTS and 1 and 10 µg/mL of PTSO. Cells incubated with PTSO obtained similar viability percentages to uninfected cells. These results suggest that the use of PTS and PTSO is a promising alternative to coccidiosis treatment, although further in vivo studies need to be performed.

Antifungal Activity of Propyl-Propane-Thiosulfinate (PTS) and Propyl-Propane-Thiosulfonate (PTSO) from Allium cepa against Verticillium dahliae: In Vitro and in Planta Assays.

Verticillium wilt, caused by Verticillium dahliae, is the most devastating soil-borne fungal disease of olive trees worldwide. Currently, there is no effective measure available to control the pathogen in diseased plants in open field conditions. Searching more effective and sustainable solutions are a priority for the olive sector. The existing alternatives for disease control include the use of biological control microorganisms and compounds of natural origin from plants, such as Alliaceae. Propyl propane thiosulfinate (PTS) and propyl propane thiosulfonate (PTSO) are two organosulfur compounds derived from Allium cepa with a widely documented antimicrobial activity. The aim of this study was to evaluate the antifungal activity of PTS and PTSO against the defoliating and non-defoliating V. dahliae pathotypes. Firstly, several in vitro tests were performed (Minimum Antifungal Concentration, susceptibility studies according to the Kirby-Bauer disk-diffusion method, antifungal activity through aerial diffusion and effect on mycelial growth). The ability of both compounds to sanitize soil was evaluated using a sterile substrate inoculated with V. dahliae. Finally, challenges in growth chambers were carried out. PTS and PTSO generated growth inhibition zones in agar diffusion and the gas phase, and the mycelial growth of all the V. dahliae strains was significantly altered. The V. dahliae population in soil was considerably reduced after the sanitization. Finally, in planta assays demonstrated the ability of these compounds to reduce disease related parameters and their contribution to control the phytopathogen. In conclusion, the results showed that the PTS and PTSO from Allium cepa display in vitro and in vivo antifungal activity against V. dahliae and suggested that both compounds could be used as natural and environmentally friendly tools for Verticillium wilt management.