Cereus jamacaru DC. (mandacaru) fruit as a source of lactic acid bacteria with in vitro probiotic-related characteristics and its protective effects on Pediococcus pentosaceus during lyophilization and refrigeration storage.

This study isolated and identified autochthonous lactic acid bacteria (LAB) from mandacaru fruit and evaluated their potential probiotic and technological aptitudes in vitro, as well as the protective effects of freeze-dried mandacaru fruit on the most promising LAB isolate during lyophilization and refrigeration storage. Initially, 212 colonies were isolated from mandacaru fruit, and 34 were preliminarily identified as LAB. Thirteen isolates identified by 16S-rRNA sequencing as Pediococcus pentosaceus were negative for DNase, gelatinase, hemolytic, and biogenic amine production. The selected isolates showed proteolytic activity, diacetyl and exopolysaccharide production, and good tolerance to different NaCl concentrations while having low cellular hydrophobicity and antagonistic activity against pathogens. The survival of isolates sharply decreased after 3 h of exposure to pH 2 and had a good tolerance to 1 % bile salt. A principal component analysis selected P. pentosaceus 57 as the most promising isolate based on the examined technological and probiotic-related physiological properties. This isolate was lyophilized with mandacaru fruit and stored under refrigeration for 90 days. P. pentosaceus 57 lyophilized with mandacaru fruit had high viable cell counts (9.69 ± 0.03 log CFU/mL) and >50 % of physiologically active cells at 90 days of refrigeration storage. The results indicate that mandacaru fruit is a source of P. pentosaceus with aptitudes to be explored as potential probiotic and technological characteristics of interest for the food industry, besides being a good candidate for use in lyophilization processes and refrigeration storage of LAB due to its cryoprotective effects.

Limosilactobacillus fermentum Strains as Novel Probiotic Candidates to Promote Host Health Benefits and Development of Biotherapeutics: A Comprehensive Review.

Fruits and their processing by-products are sources of potentially probiotic strains. Limosilactobacillus (L.) fermentum strains isolated from fruit processing by-products have shown probiotic-related properties. This review presents and discusses the results of the available studies that evaluated the probiotic properties of L. fermentum in promoting host health benefits, their application by the food industry, and the development of biotherapeutics. The results showed that administration of L. fermentum for 4 to 8 weeks promoted host health benefits in rats, including the modulation of gut microbiota, improvement of metabolic parameters, and antihypertensive, antioxidant, and anti-inflammatory effects. The results also showed the relevance of L. fermentum strains for application in the food industry and for the formulation of novel biotherapeutics, especially nutraceuticals. This review provides evidence that L. fermentum strains isolated from fruit processing by-products have great potential for promoting host health and indicate the need for a translational approach to confirm their effects in humans using randomized, double-blind, placebo-controlled trials.

Unrevealing the in vitro impacts of Cereus jacamaru DC. cladodes flour on potentially probiotic strains, selected bacterial populations, and metabolic activity of human intestinal microbiota.

This study investigated the potential impacts of the flour from Cereus jamacaru cactus cladodes (CJF), a cactus native to the Brazilian Caatinga biome, on the growth and metabolism of different potentially probiotic strains, as well as on the abundance of selected intestinal bacterial populations and microbial metabolic activity during in vitro colonic fermentation with a pooled human fecal inoculum. Cultivation of the probiotics in a medium with C. jamacaru cladodes flour (20 g/L) resulted in viable cell counts of up to 9.8 log CFU/mL, positive prebiotic activity scores (0.73-0.91), decreased pH and sugar contents, and increased lactic, acetic, and propionic acid production over time, indicating enhanced probiotic growth and metabolic activity. CJF overall increased the relative abundance of Lactobacillus spp./Enterococcus spp. (2.12-3.29%) and Bifidobacterium spp. (4.08-4.32%) and decreased the relative abundance of Bacteroides spp./Prevotella spp. (8.35-6.81%), Clostridium histolyticum (6.91-3.59%), and Eubacterium rectale/Clostridium coccoides (7.70-3.95%) during 48 h of an in vitro colonic fermentation using a pooled human fecal inoculum. CJF stimulated the microbial metabolic activity, with decreased pH, sugar consumption, lactic and short-chain fatty acid production, alterations in overall metabolic profiling and phenolic compound contents, and maintenance of high antioxidant capacity during colonic fermentation. These results show that CJF stimulated the growth and metabolic activity of distinct potential probiotics, increased the relative abundance of beneficial intestinal bacterial groups, and stimulated microbial metabolism during in vitro colonic fermentation. Further studies using advanced molecular technologies and in vivo experimental models could forward the investigation of the potential prebiotic properties of CJF.

An In Vitro Study of the Impacts of Sweet Potato Chips with Potentially Probiotic Levilactobacillus brevis and Lactiplantibacillus plantarum on Human Intestinal Microbiota : Impacts of potato chips with probiotics on intestinal microbiota.

This study formulated sweet potato chips with powdered potentially probiotic Levilactobacillus brevis (SPLB) and Lactiplantibacillus plantarum (SPLP) and evaluated their impacts on human intestinal microbiota during 48 h of in vitro colonic fermentation. L. brevis and L. plantarum kept high viable cell counts (> 6 log CFU/g) on sweet potato chips after freeze-drying and during 60 days of storage. SPLB and SPLP had satisfactory quality parameters during 60 days of storage. SPLB and SPLP increased the relative abundance of Lactobacillus ssp./Enterococcus spp. (3.84-10.22%) and Bifidobacterium spp. (3.25-12.45%) and decreased the relative abundance of Bacteroides spp./Prevotella spp. (8.56-2.16%), Clostridium histolyticum (8.23-2.33%), and Eubacterium rectale/Clostridium coccoides (8.07-1.33%) during 48 h of in vitro colonic fermentation. SPLB and SPLP achieved high positive prebiotic indexes (> 8.24), decreased pH values and sugar contents, and increased lactic acid and short-chain fatty acid production, proving selective stimulatory effects on beneficial bacterial groups forming the intestinal microbiota. The results showed that SPLB and SPLP have good stability and high viable cell counts of L. brevis and L. plantarum when stored under room temperature and caused positive impacts on human intestinal microbiota, making them potentially probiotic non-dairy snack options.

Revealing the Potential Impacts of Nutraceuticals Formulated with Freeze-Dried Jabuticaba Peel and Limosilactobacillus fermentum Strains Candidates for Probiotic Use on Human Intestinal Microbiota.

This study evaluated the impacts of novel nutraceuticals formulated with freeze-dried jabuticaba peel (FJP) and three potentially probiotic Limosilactobacillus fermentum strains on the abundance of bacterial groups forming the human intestinal microbiota, metabolite production, and antioxidant capacity during in vitro colonic fermentation. The nutraceuticals had high viable counts of L. fermentum after freeze-drying (≥ 9.57 ± 0.09 log CFU/g). The nutraceuticals increased the abundance of Lactobacillus ssp./Enterococcus spp. (2.46-3.94%), Bifidobacterium spp. (2.28-3.02%), and Ruminococcus albus/R. flavefaciens (0.63-4.03%), while decreasing the abundance of Bacteroides spp./Prevotella spp. (3.91-2.02%), Clostridium histolyticum (1.69-0.40%), and Eubacterium rectale/C. coccoides (3.32-1.08%), which were linked to positive prebiotic indices (> 1.75). The nutraceuticals reduced the pH and increased the sugar consumption, short-chain fatty acid production, phenolic acid content, and antioxidant capacity, besides altering the metabolic profile during colonic fermentation. The combination of FJP and probiotic L. fermentum is a promising strategy to produce nutraceuticals targeting intestinal microbiota.

Investigating the effects of conventional and unconventional edible parts of red beet (Beta vulgaris L.) on target bacterial groups and metabolic activity of human colonic microbiota to produce novel and sustainable prebiotic ingredients.

This study investigated the effects of freeze-dried red beet root (FDBR) and freeze-dried red beet stem and leaves (FDBSL) on target bacterial groups and metabolic activity of human colonic microbiota in vitro. The capability of FDBR and FDBSL to cause alterations in the relative abundance of different selected bacterial groups found as part of human intestinal microbiota, as well as in pH values, sugar, short-chain fatty acid, phenolic compounds, and antioxidant capacity were evaluated during 48 h of in vitro colonic fermentation. FDBR and FDBSL were submitted to simulated gastrointestinal digestion and freeze-dried prior to use in colonic fermentation. FDBR and FDBSL overall increased the relative abundance of Lactobacillus spp./Enterococcus spp. (3.64-7.60%) and Bifidobacterium spp. (2.76-5.78%) and decreased the relative abundance of Bacteroides spp./Prevotella spp. (9.56-4.18%), Clostridium histolyticum (1.62-1.15%), and Eubacterium rectale/Clostridium coccoides (2.33-1.49%) during 48 h of colonic fermentation. FDBR and FDBSL had high positive prebiotic indexes (>3.61) during colonic fermentation, indicating selective stimulatory effects on beneficial intestinal bacterial groups. FDBR and FDBSL increased the metabolic activity of human colonic microbiota, evidenced by decreased pH, sugar consumption, short-chain fatty acid production, alterations in phenolic compound contents, and maintenance of high antioxidant capacity during colonic fermentation. The results indicate that FDBR and FDBSL could induce beneficial alterations in the composition and metabolic activity of human intestinal microbiota, as well as that conventional and unconventional red beet edible parts are candidates to use as novel and sustainable prebiotic ingredients.

Survival kinetics, membrane integrity and metabolic activity of Salmonella enterica in conventionally and osmotically dehydrated coconut flakes.

Many outbreaks involving Salmonella enterica in dehydrated coconut have been reported. Little is known about the survival of S. enterica in dehydrated coconut flakes at common retail or domestic storage conditions. This study evaluated the behavior of a S. enterica cocktail (S. Enteritidis PT4, S. Typhimurium PT4, S. Bredeney, S. Muenster and S. Agona) in conventionally and osmotically dehydrated coconut flakes under four storage regimes: 25 °C for 120 days, 25 °C for 30 days then 7 °C for 90 days, 7 °C for 30 days then 25 °C for 90 days, and 7 °C for 120 days. S. enterica membrane integrity (using with propidium iodide and bis-1,3-dibutylbarbutyric acid) and metabolic activity (using 5-cyano-2,3-ditolyl tetrazolium chloride) were assessed by flow cytometry analysis after dehydration and storage at 7 °C or 25 °C for 120 days. Lower S. enterica inactivation rates (kmax 0.02 to 0.04 1/days) were observed in conventionally dehydrated coconut flakes compared to osmotically dehydrated coconut flakes (kmax 0.16 to 0.20 1/days). Changes in storage temperature did not affect the behavior of S. enterica in conventionally or osmotically dehydrated coconut flakes. Results show that S. enterica inactivation in conventionally dehydrated coconut flakes could be described by log-linear with tail models. S. enterica inactivation in osmotically dehydrated coconut flakes could be described by log-linear with shoulder and tail models. Subpopulations of S. enterica cells with damaged membranes and without metabolic activity were larger in conventionally (32.1% and 90.9%, respectively) than osmotically dehydrated coconut (18.5% and 82.2%, respectively) flakes at the beginning of the storage. Subpopulations of S. enterica cells with damaged membrane decreased by 9.4-14.4%, while cells with membrane potential and intact membrane increased by 23.7 and 24.2% in conventionally dehydrated coconut flakes after 120 days of storage at 7 °C or 25 °C, respectively. Subpopulations of S. enterica with damaged membranes did not change significantly in osmotically dehydrated coconut flakes. Our findings suggest that S. enterica populations decline during storage occurs due in part to membrane integrity losses. These data can contribute to the development of risk management strategies for S. enterica in dehydrated coconut flakes.

Antifungal effects of Conyza bonariensis (L.) Cronquist essential oil against pathogenic Colletotrichum musae and its incorporation in gum Arabic coating to reduce anthracnose development in banana during storage.

AIM: This study evaluated the inhibitory effects on mycelial growth and damage on membrane integrity and enzymatic activity caused by Conyza bonariensis essential oil (CBEO) on distinct pathogenic Colletotrichum musae isolates, as well as the preventive and curative effects of coatings with gum Arabic (GA) and CBEO to reduce anthracnose development in banana during room temperature storage. The effects of GA-CBEO coatings on some physicochemical parameters of banana were investigated during room temperature storage. METHOD AND RESULTS: CBEO (0.4-1 µl ml-1 ) inhibited the mycelial growth of C. musae isolates in laboratory media. The exposure of C. musae conidia to CBEO (0.6 µl ml-1 ) for 3 and 5 days resulted in high percentages of conidia with damaged cytoplasmic membrane and without enzymatic activity. Coatings with GA (0.1 mg ml-1 ) and CBEO (0.4-1 µl ml-1 ) reduced the anthracnose development in banana artificially contaminated with C. musae during storage. In most cases, the disease severity indexes found for GA-CBEO-coated banana were lower than or similar to those for banana treated with commercial fungicide. GA-CBEO-coated banana had reduced alterations in physicochemical parameters during storage, indicating more prolonged storability. CONCLUSION: The application of GA-CBEO coatings is effective to delay the anthracnose development in banana during storage, which should help to reduce the amount of fungicides used to control postharvest diseases in this fruit. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study showing the efficacy of coatings formulated with GA and CBEO to delay the development of anthracnose in banana, as well as to decrease alterations in physicochemical parameters indicative of postharvest quality of this fruit during storage. In a practical point of view, GA-CBEO coatings could be innovative strategies to delay the anthracnose development and postharvest losses in banana.

Selection of Lactic Acid Bacteria with In Vitro Probiotic-Related Characteristics from the Cactus Pilosocereus gounellei (A. Weber ex. K. Schum.) Bly. ex Rowl.

Pilosocereus gounellei (A. Weber ex. K. Schum.) Bly. ex Rowl., popularly known as xique-xique, is a cactus from the Caatinga biome, which is rich in bioactive compounds but has not been previously studied as a source of lactic acid bacteria (LAB) with probiotic aptitudes. This study aimed to identify, characterize, and select LAB isolates with in vitro probiotic-related characteristics from xique-xique cladodes and fruit. Isolates with the most promising probiotic-related characteristics were evaluated regarding their in vitro technological properties and capability of surviving in chestnut milk, whey protein drink, and mate tea with mint during 21 days of refrigeration storage. Seventeen recovered isolates had typical characteristics of LAB. Six out of these seventeen LAB isolates passed the safety tests and were included in experiments to evaluate the in vitro probiotic-related characteristics. Based on the results of a principal component analysis, the isolates 69, 82, 98, and 108 had the best performances in experiments to evaluate the probiotic-related characteristics. In addition to showing good technological properties, the four selected LAB isolates had high viable counts (>7.3 log cfu/mL) and high sizes of physiologically active cell subpopulations in chestnut milk, whey protein drink, and mate tea during refrigeration storage. These four isolates were identified by 16S-rRNA sequencing as being Lacticaseibacillus paracasei or Lacticaseibacillus casei. The results indicate xique-xique as a source of potentially probiotic LAB isolates.

Effects of digested flours from four different sweet potato (Ipomoea batatas L.) root varieties on the composition and metabolic activity of human colonic microbiota in vitro.

This study evaluated the effects of flours from four different sweet potato root (SPR) varieties, being two with white peel and two with purple peel, on the composition and metabolic activity of human colonic microbiota in vitro. The capability of these SPR flours (20 g/L) to cause alterations in relative abundance of different bacterial groups found as part of human colonic microbiota, as well as in lactic acid and short-chain fatty acid production was evaluated during 48 hr of an in vitro colonic fermentation. The SPR flours were submitted to a simulated gastrointestinal digestion prior to use in experiments. The four SPR flours increased the relative abundance of Lactobacillus/Enterococcus (range: 0.49-4.48%) and Bifidobacterium (range: 0.32-3.27%) and decreased the relative abundance of Bacteroides/Prevotella (range: 0.29-7.49%), Clostridium histolyticum (range: 0.15-2.08%), and Eubacterium rectale/Clostridium coccoides (range: 0.28-3.86%) during the 48 hr of colonic fermentation. The four SPRF flours had positive prebiotic indexes (> 0.38) after 24 and 48 hr of colonic fermentation, reinforcing the occurrence of selective stimulatory effects on colonic microbiota. An increased metabolic activity of human colonic microbiota was caused by tested SPR flours, which was evidenced by decreased pH (range: 3.20-3.83) and increased lactic acid and short chain fatty acid production during the 48 hr of colonic fermentation. The four examined SPR flours were capable of causing positive alterations in composition and driving the metabolic activity of human colonic microbiota during in vitro colonic fermentation, which should be linked to their prebiotic properties. PRACTICAL APPLICATION: The four examined sweet potato root flours (SPRF) caused beneficial alterations in composition besides of driving the metabolic activity of human colonic microbiota in vitro. These results characterize the examined SPRF as candidates for use as prebiotic ingredients by food industry for formulation of value-added functional foods or dietary supplements.

Effects of Quercetin and Resveratrol on in vitro Properties Related to the Functionality of Potentially Probiotic Lactobacillus Strains.

The ability of probiotics to exert benefits on host has been associated with different physiological functionalities in these microorganisms, namely cell surface hydrophobicity, autoaggregation, coaggregation with pathogens, antagonistic activity against pathogens and ability to survive the exposure to gastrointestinal conditions. This study assessed the effects of different concentrations of quercetin (QUE) and resveratrol (RES) on the ability of six potentially probiotic Lactobacillus strains to tolerate different pH values and bile salt concentrations, to autoaggregate, coaggregate with and antagonize pathogens and survive the exposure to simulated gastrointestinal conditions. QUE and RES presented low inhibitory effects on all tested Lactobacillus strains, with minimum inhibitory concentration (MIC) ranging from 512 to >1024 µg/mL. In most cases, QUE and RES at all tested concentrations (i.e., MIC, 1/2 MIC, and 1/4 MIC) did not affect the tolerance of the Lactobacillus strains to acidic pH and bile salts. QUE increased cell surface hydrophobicity of most of the tested Lactobacillus strains and increases or decreases in cell surface hydrophobicity varied in the presence of different RES concentrations among some strains. QUE and RES increased the ability of tested Lactobacillus strains to autoaggregate and coaggregate with pathogens. QUE and RES did not negatively affect the antagonistic activity of the tested Lactobacillus strains against pathogens and did not decrease their survival rates when exposed to in vitro gastrointestinal conditions. In a few cases, the ability of some tested Lactobacillus strains to antagonize pathogens, as well as to survive specific steps of the in vitro digestion was increased by QUE and RES. QUE exerted overall better protective effects on the measured in vitro properties of tested Lactobacillus strains than RES, and L. fermentum and L. plantarum strains presented better responses when treated with QUE or RES. These results showed that probiotic Lactobacillus strains could present low susceptibility to QUE and RES. Combined use of QUE and RES with probiotic Lactobacillus could improve their functionalities on the host; however, the concentration of these polyphenols should be carefully selected to achieve the desirable effects and vary according to the selected probiotic strain.

Potential interactions among phenolic compounds and probiotics for mutual boosting of their health-promoting properties and food functionalities - A review.

Several foods are rich sources of phenolic compounds (PC) and their beneficial effects on human health may be increased through the action of probiotics. Additionally, probiotics may use PC as substrates, increasing their survival and functionality. This review presents available studies on the effects of PC on probiotics, including their physiological functionalities, interactions and capability of surviving during exposure to gastrointestinal conditions and when incorporated into food matrices. Studies have shown that PC can improve the adhesion capacity and survival of probiotics during exposure to conditions that mimic the gastrointestinal tract. There is strong evidence that PC can modulate the composition of the gut microbiota in hosts, improving a variety of biochemical markers and risk factors for chronic diseases. Available literature also indicates that metabolites of PC formed by intestinal microorganisms, including probiotics, exert a variety of benefits on host health. These metabolites are typically more active than parental dietary PC. The presence of PC commonly enhances probiotic survival in different foods. Finally, further clinical studies need to be developed to confirm in vitro and experimental findings concerning the beneficial interactions among different PC and probiotics.

The performance of five fruit-derived and freeze-dried potentially probiotic Lactobacillus strains in apple, orange, and grape juices.

BACKGROUND: This study assessed the survival of the fruit-derived and freeze-dried L. plantarum 49, L. brevis 59, L. paracasei 108, L. fermentum 111 and L. pentosus 129 strains during frozen storage and when incorporated into apple, orange, and grape juice stored under refrigeration. Physicochemical parameters of juices containing the freeze-dried Lactobacillus strains and the survival of the test strains in the fruit juices during in vitro digestion were also evaluated. RESULTS: No decreases in survival rates (log N/log N0) of the freeze-dried cells were observed in up to 1 month of storage. The survival rates of the freeze-dried strains L. plantarum 49 and L. paracasei 108 were > 0.75 in up to 4 months of storage. All freeze-dried strains exhibited survival rates of >0.75 in up to 2 weeks of storage in apple juice; only L. plantarum 49 and L. paracasei 108 showed similar survival rates in orange and grape juices in up to 2 weeks of storage. The contents of the monitored organic acids or sugars during storage varied depending on the added strain and the type of fruit juice. At the end of in vitro digestion, L. brevis 59, L. paracasei 108 and L. fermentum 111 showed survival rates of >0.80 in apple juice. CONCLUSION: Apple juice was the best substrate for the survival of the tested freeze-dried Lactobacillus strains over time. L. paracasei 108 and L. plantarum 49 were the strains presenting the best performance for incorporation in potentially probiotic fruit juices. © 2018 Society of Chemical Industry.

In Vitro Characterization of Lactobacillus Strains Isolated from Fruit Processing By-Products as Potential Probiotics.

Nine wild Lactobacillus strains, namely Lactobacillus plantarum 53, Lactobacillus fermentum 56, L. fermentum 60, Lactobacillus paracasei 106, L. fermentum 250, L. fermentum 263, L. fermentum 139, L. fermentum 141, and L. fermentum 296, isolated from fruit processing by-products were evaluated in vitro for a series of safety, physiological functionality, and technological properties that could enable their use as probiotics. Considering the safety aspects, the resistance to antibiotics varied among the examined strains, and none of the strains presented hemolytic and mucinolytic activity. Regarding the physiological functionality properties, none of the strains were able to deconjugate bile salts; all of them presented low to moderate cell hydrophobicity and were able to autoaggregate, coaggregate with Listeria monocytogenes and Escherichia coli, and antagonize pathogenic bacteria. Exposure to pH 2 sharply decreased the survival of the examined strains after 1- or 2-h exposure; variable decreases were noted after 3-h exposure to pH 3. Overall, exposure to pH 5 and to bile salts (0.15, 0.3, and 1%) did not decrease the strains' survival. Examined strains presented better ability to survive from the exposure to simulated gastrointestinal conditions in laboratorial media and milk than in grape juice. Considering the technological properties, all the strains were positive for proteolytic activity and EPS and diacetyl production, and most of them had good tolerance to 1-4% NaCl. These results indicate that wild Lactobacillus strains isolated from fruit processing by-products could present performance compatible with probiotic properties and technological features that enable the development of probiotic foods with distinct characteristics.