A Mix of Potentially Probiotic Limosilactobacillus fermentum Strains Alters the Gut Microbiota in a Dose- and Sex-Dependent Manner in Wistar Rats.

Multi-strain Limosilactobacillus (L.) fermentum is a potential probiotic with reported immunomodulatory properties. This study aimed to evaluate the composition, richness, and diversity of the gut microbiota in male and female rats after treatment with a multi-strain of L. fermentum at different doses. Thirty rats (fifteen male and fifteen female) were allocated into a control group (CTL), a group receiving L. fermentum at a dose of 108 CFU (Lf-108), and a group receiving L. fermentum at a dose of 1010 CFU (Lf-1010) for 13 weeks. Gut microbiota and serum cytokine levels were evaluated after L. fermentum treatment. Male CTL rats had a lower relative abundance of Bifidobacteriaceae and Prevotella and a lower alpha diversity than their female CTL counterparts (p < 0.05). In addition, male CTL rats had a higher Firmicutes/Bacteroidetes (F/B) ratio than female CTL rats (p < 0.05). In female rats, the administration of L. fermentum at 108 CFU decreased the relative abundance of Bifidobacteriaceae and Anaerobiospirillum and increased Lactobacillus (p < 0.05). In male rats, the administration of L. fermentum at 1010 CFU decreased the F/B ratio and increased Lachnospiraceae and the diversity of the gut microbiota (p < 0.05). The relative abundance of Lachnospiraceae and the alpha-diversity of gut microbiota were negatively correlated with serum levels of IL1ß (r = -0.44) and TNFα (r = -0.39), respectively. This study identified important changes in gut microbiota between male and female rats and showed that a lower dose of L. fermentum may have more beneficial effects on gut microbiota in females, while a higher dose may result in more beneficial effects on gut microbiota in male rats.

Safety Evaluation of a Novel Potentially Probiotic Limosilactobacillus fermentum in Rats.

Limosilactobacillus (L) fermentum (strains 139, 263, 296) is a novel probiotic mixture isolated from fruit processing by-products. The use of this formulation has been associated with improvements in cardiometabolic, inflammatory, and oxidative stress parameters. The present study evaluated the safety of a potential multi-strain probiotic by genotoxicity (micronucleus assay) and subchronic toxicity study (13-week repeated dose). In the genotoxicity evaluation, L. fermentum 139, 263, 296 did not increase the frequency of micronuclei in erythrocytes of rats of both sexes at doses up to 1010 CFU/mL. In the subchronic toxicity study, the administration of L. fermentum did not promote adverse health effects, such as behavioral changes, appearance of tumors, changes in hematological and biochemical parameters. In addition, higher doses of L. fermentum 139, 263, 296 have been shown to reduce the levels of pro-inflammatory cytokines. Administration of potentially probiotic L. fermentum did not promote adverse health effects in rats and could be evaluated as a potential probiotic for humans.

Limosilactobacillus fermentum Strains with Claimed Probiotic Properties Exert Anti-oxidant and Anti-inflammatory Properties and Prevent Cardiometabolic Disorder in Female Rats Fed a High-Fat Diet.

This study assessed the effects of a mixed formulation containing Limosilactobacillus (L.) fermentum 139, L. fermentum 263, and L. fermentum 296 on cardiometabolic parameters, inflammatory markers, short-chain fatty acid (SCFA) fecal contents, and oxidative stress in colon, liver, heart, and kidney tissues of female rats fed a high-fat diet (HFD). Female Wistar rats were allocated into control diet (CTL, n = 6), HFD (n = 6), and HFD receiving L. fermentum formulation (HFD-LF, n = 6). L. fermentum formulation (1 × 109 CFU/mL of each strain) was administered two twice a day for 4 weeks. Administration of L. fermentum increased acetate and succinate fecal contents and reduced hyperlipidemia and hyperglycemia in rats fed a HFD (p < 0.05). Administration of L. fermentum decreased low-grade inflammation and improved antioxidant capacity along the gut, liver, heart, and kidney tissues in female rats fed a HFD (p < 0.05). Administration of L. fermentum prevented dyslipidemia, inflammation, and oxidative stress in colon, liver, heart, and kidney in female rats fed a HFD.

Application of Potentially Probiotic Fruit-Derived Lactic Acid Bacteria Loaded into Sodium Alginate Coatings to Control Anthracnose Development in Guava and Mango During Storage.

This study evaluated the efficacy of potentially probiotic fruit-derived lactic acid bacteria (LAB) strains loaded into sodium alginate (SA) coatings to control the anthracnose development in guava cv. Paluma and mango cv. Palmer caused by distinct pathogenic Colletotrichum species (C. asianum, C. fructicola, C. tropicale, C. siamense, C. karstii, and C. gloeosporioides) during 15 days of room temperature storage (25 ± 0.5 °C). The effects of the formulated coatings on physicochemical parameters indicative of overall postharvest quality of guava and mango were evaluated. The eight examined LAB strains caused strong inhibition on the mycelial growth of all target Colletotrichum species in vitro. LAB strains with the highest inhibitory effects (Levilactobacillus brevis 59, Lactiplantibacillus pentosus 129, and Limosilactobacillus fermentum 263) on the target Colletotrichum species were incorporated into SA coatings. These strains had viable counts of > 6 log CFU/mL in SA coatings during 15 days of room temperature storage. Application of coatings with SA + L. brevis 59, SA + L. pentosus 129, and SA + L. fermentum 263 delayed the development and decreased the severity of anthracnose lesions in guava and mango artificially contaminated with either of the tested Colletotrichum species. These coatings impacted positively on some physicochemical parameters indicative of postharvest quality and more prolonged storability of guava and mango. The formulated SA coatings loaded with tested fruit-derived potentially probiotic LAB strains could be innovative and effective strategies to control postharvest anthracnose and extend the storability of guava and mango.

Effects of a Mixed Limosilactobacillus fermentum Formulation with Claimed Probiotic Properties on Cardiometabolic Variables, Biomarkers of Inflammation and Oxidative Stress in Male Rats Fed a High-Fat Diet.

High-fat diet (HFD) consumption has been linked to dyslipidemia, low-grade inflammation and oxidative stress. This study investigated the effects of a mixed formulation with Limosilactobacillusfermentum 139, L. fermentum 263 and L. fermentum 296 on cardiometabolic parameters, fecal short-chain fatty acid (SCFA) contents and biomarkers of inflammation and oxidative stress in colon and heart tissues of male rats fed an HFD. Male Wistar rats were grouped into control diet (CTL, n = 6), HFD (n = 6) and HFD with L. fermentum formulation (HFD-Lf, n = 6) groups. The L.fermentum formulation (1 × 109 CFU/mL of each strain) was administered twice a day for 4 weeks. After a 4-week follow-up, biochemical parameters, fecal SCFA, cytokines and oxidative stress variables were evaluated. HFD consumption caused hyperlipidemia, hyperglycemia, low-grade inflammation, reduced fecal acetate and propionate contents and increased biomarkers of oxidative stress in colon and heart tissues when compared to the CTL group. Rats receiving the L. fermentum formulation had reduced hyperlipidemia and hyperglycemia, but similar SCFA contents in comparison with the HFD group (p < 0.05). Rats receiving the L. fermentum formulation had increased antioxidant capacity throughout the colon and heart tissues when compared with the control group. Administration of a mixed L. fermentum formulation prevented hyperlipidemia, inflammation and oxidative stress in colon and heart tissues induced by HFD consumption.

Current Advances on the Development and Application of Probiotic-Loaded Edible Films and Coatings for the Bioprotection of Fresh and Minimally Processed Fruit and Vegetables.

The application of probiotics has emerged as an innovative bioprotection technology to preserve fresh and minimally processed fruit and vegetables. This review discusses the most recent advances on the development and application of probiotic-loaded edible films/coatings as a strategy to preserve fresh or minimally processed fruit and vegetables. Available studies have shown a variety of materials, including hydrocolloids (polysaccharides and proteins) and lipids, used alone or in combination to formulate edible films/coatings loaded with probiotics. Plasticizers and surfactants are usually required to formulate these edible films/coatings. The reported antimicrobial effects of probiotic-loaded edible films/coating and quality parameters of coated fruit and vegetables could vary according to the characteristics of the materials used in their formulation, loaded probiotic strain and its dose. The antimicrobial effects of these films/coatings could be linked to the action of various metabolites produced by embedded probiotic cells with inhibitory effects on microorganisms contaminating fruit and vegetable surfaces. The implication of the use of probiotic-loaded edible films/coatings should be their antimicrobial effects against pathogenic and spoilage microorganisms and efficacy to control the ripening of fruit and vegetables, helping the coated products to maintain their safety, quality, nutritional and functional characteristics for a more prolonged storage period.

Coatings with chitosan and phenolic-rich extract from acerola (Malpighia emarginata D.C.) or jabuticaba (Plinia jaboticaba (Vell.) Berg) processing by-product to control rot caused by Lasiodiplodia spp. in papaya (Carica papaya L.) fruit.

This study evaluated if coatings with chitosan (Chi) and phenolic-rich extract from acerola (Malpighia emarginata D.C., PEA) or jabuticaba (Plinia jaboticaba (Vell.) Berg, PEJ) processing by-products are effective to control the development of rot caused by Lasiodiplodia pseudotheobromae, L. viticola, L. euphorbicola, L. theobromae and L. hormozganensis in papaya (Carica papaya L.) fruit. Effects of formulated coatings on some physicochemical parameters indicative of postharvest quality of papaya were investigated. Twenty-six different phenolics were found in PEA and PEJ, including flavonoids, stilbenes, tannins and phenolic acids. Chi (1-5 mg/mL), PEA and PEJ (25-100 mg/mL) separately caused mycelial growth inhibition on all isolates. Combinations of Chi (3 and 4 mg/mL) and PEA (50 and 75 mg/mL) or PEJ (75 and 100 mg/mL) had additive interactions. Coatings with Chi (4 mg/mL) and PEA (50 or 75 mg/mL) or PEA (75 or 100 mg/mL) inhibited rot development in papaya fruit infected with Lasiodiplodia isolates during 8 days of room temperature storage. Coatings with 4 mg/mL Chi and 75 mg/mL PEA or 100 mg/mL PEJ were the most effective to control rot development. These coatings did not affect negatively physicochemical parameters indicative of postharvest quality of papaya fruit during storage. Coatings with combined Chi and PEA or PEJ could be novel strategies to control postharvest rot caused by Lasiodiplodia in papaya fruit.

Chitosan produced from Mucorales fungi using agroindustrial by-products and its efficacy to inhibit Colletotrichum species.

This study evaluated corn steep liquor (CSL) and papaya peel juice (PPJ) in mixture as substrates for the cultivation (96h, 28°C, pH 5.6, 150rpm) of Mucorales fungi for chitosan production, and determined the growth-inhibitory effect of the fungal chitosan (FuCS) obtained under optimized conditions against phytopathogenic Colletotrichum species. All Mucorales fungi tested were capable of growing in CSL-PPJ medium, showing FuCS production in the range of 5.02 (Fennelomyces heterothalicus SIS 28) - 15.63mg/g (Cunninghamella elegans SIS 41). Highest FuCS production (37.25mg/g) was achieved when C. elegans was cultivated in medium containing 9.43% CSL and 42.5% PPJ. FuCS obtained under these conditions showed a deacetylation degree of 86%, viscosity of 120cP and molecular weight of 4.08×104g/mol. FuCS at 5000, 7500 and 10,000ppm inhibited the growth of all Colletotrichum species tested. FuCS also induced alterations in the morphology of C. fructicola hyphae. CSL-PPJ mixtures are suitable substrates for the cultivation of Mucorales fungi for FuCS production. Chitosan from C. elegans cultivated in CSL-PPJ medium is effective in inhibiting phytopathogenic Colletotrichum species.

Control of anthracnose caused by Colletotrichum species in guava, mango and papaya using synergistic combinations of chitosan and Cymbopogon citratus (D.C. ex Nees) Stapf. essential oil.

This study assessed the efficacy of chitosan (Chi) and Cymbopogon citratus (D.C. ex Nees) Stapf. essential oil (CCEO) combinations to control the mycelial growth of five pathogenic Colletotrichum species (C. asianum, C. siamense, C. fructicola, C. tropicale and C. karstii) in vitro, as well as the anthracnose development in guava (Psidium guajava L.) cv. Paluma, mango (Mangifera indica L.) cv. Tommy Atkins and papaya (Carica papaya L.) cv. Papaya artificially inoculated with these species. Combinations of Chi (2.5, 5 or 7.5mg/mL) and CCEO (0.15, 0.3, 0.6 or 1.25µL/mL) inhibited the mycelial growth of all tested fungal species in vitro. Examined Chi-CCEO combinations showed additive or synergistic interactions to inhibit the target Colletotrichum species based on the Abbott index. Coatings formed by synergistic Chi (5mg/mL) and CCEO (0.15, 0.3 or 0.6µL/mL) combinations decreased anthracnose lesion development in guava, mango and papaya inoculated with any of the tested Colleotrichum species during storage. Overall, anthracnose lesion development inhibition in fruit coated with synergistic Chi-CCEO combinations was higher than that observed in fruit treated with synthetic fungicides. These results show that the application of coatings formed by Chi-CCEO synergistic combinations could be effective to control postharvest anthracnose development in fruit.

Synergistic mixtures of chitosan and Mentha piperita L. essential oil to inhibit Colletotrichum species and anthracnose development in mango cultivar Tommy Atkins.

This study assessed the efficacy of chitosan (CHI) and Mentha piperita L. essential oil (MPEO) alone or in combination to control the mycelial growth of five different Colletotrichum species, C. asianum, C. dianesei, C. fructicola, C. tropicale and C. karstii, identified as potential anthracnose-causing agents in mango (Mangifera indica L.). The efficacy of coatings of CHI and MPEO mixtures in controlling the development of anthracnose in mango cultivar Tommy Atkins was evaluated. CHI (2.5, 5, 7.5 and 10 mg/mL) and MPEO (0.3, 0.6, 1.25, 2.5 and 5 µL/mL) alone effectively inhibited mycelial growth of all tested Colletotrichum strains in synthetic media. Mixtures of CHI (5 or 7.5 mg/mL) and MPEO (0.3, 0.6 or 1.25 µL/mL) strongly inhibited mycelial growth and showed additive or synergistic inhibitory effects on the tested Colletotrichum strains based on the Abbott index. The application of coatings of CHI (5 or 7.5 mg/mL) and MPEO (0.6 or 1.25 µL/mL) mixtures that presented synergistic interactions decreased anthracnose lesion severity in mango artificially contaminated with either of the tested Colletotrichum strains over 15 days of storage at 25 °C. The anthracnose lesion severity in mango coated with the mixtures of CHI and MPEO was similar or lower than those observed in mango treated with the synthetic fungicides thiophanate-methyl (10 µg a.i./mL) and difenoconazole (0.5 µg a.i./mL). The application of coatings containing low doses of CHI and MPEO may be an effective alternative for controlling the postharvest development of anthracnose in mango cultivar Tommy Atkins.

Influence of carvacrol and 1,8-cineole on cell viability, membrane integrity, and morphology of Aeromonas hydrophila cultivated in a vegetable-based broth.

This study investigated the effects of carvacrol (CAR) and 1,8-cineole (CIN) alone (at the MIC) or in combination at subinhibitory amounts (both at 1/8 MIC) on the cell viability, membrane permeability, and morphology of Aeromonas hydrophila INCQS 7966 (A. hydrophila) cultivated in a vegetable-based broth. CAR and CIN alone or in combination severely affected the viability of the bacteria and caused dramatic changes in the cell membrane permeability, leading to cell death, as observed by confocal laser microscopy. Scanning and transmission electron microscopy images of bacterial cells exposed to CAR or CIN or the mixture of both compounds revealed severe changes in cell wall structure, rupture of the plasma membrane, shrinking of cells, condensation of cytoplasmic content, leakage of intracellular material, and cell collapse. These findings suggest that CAR and CIN alone or in combination at subinhibitory amounts could be applied to inhibit the growth of A. hydrophila in foods, particularly as sanitizing agents in vegetables.