Wheat-durum pasta added of inactivated Bifidobacterium animalis decreases glucose and total cholesterol levels and modulates gut microbiota in healthy rats.

This study investigated the effects of consumption of wheat-durum pasta added of Bifidobacterium animalis inactivated by gamma-irradiation (paraprobiotic) on health and gut microbiota of rats. Twenty-one male rats were divided into three groups as follow: control (Control) receiving standard diet; pasta control (PC) receiving diet containing pasta, and paraprobiotic pasta Bifidobacterium-irradiated (PPBI), receiving paraprobiotic pasta. The serum levels of glucose and total cholesterol were reduced (p < 0.05) in PPBI when compared to the Control (p < 0.05). PPBI showed high abundance (p ˂ 0.05) of Firmicutes and Actinobacteria and a lower abundance of Bacteriodes compared to Control and PC. Besides, the PPBI showed high abundance (p ˂ 0.05) of Clostridiales, Lactobacillales; Bifidobacteriales, Bacillales, and Coriobacteriales in comparison to Control. Specific genera such as Bifibacterium, Lactobacillus, Ruminococcus, Allobaculum, and Blautia were more related to PPBI. Findings reveal wheat-durum pasta as a potential vehicle for delivering paraprobiotics B. animalis.

Paraprobiotics obtained by six different inactivation processes: impacts on the biochemical parameters and intestinal microbiota of Wistar male rats.

This study investigated the effects of feeding paraprobiotics obtained by six processes [heat, ultrasound, high pH, low pH, irradiation and supercritical carbon dioxide (CO2)] on biochemical parameters and intestinal microbiota of Wistar male rats. Daily administration of paraprobiotics did not affect (p ≥ 0.05) the food intake, body weight, glucose and triglycerides levels, expression of antioxidant enzymes or thermal shock proteins in comparison to the control. Bifidobacterium lactis inactivated by irradiation and supercritical CO2 decreased the total cholesterol levels in serum (p < 0.05). Bifidobacterium lactis inactivated by supercritical CO2 increased the albumin and creatinine levels, while decreased the HDL-cholesterol levels (p < 0.05). Clostridiales (45.6-56%), Bacteroidales (31.9-44.2%) and Lactobacillales (3.9-7.8%) corresponded to the major orders in paraprobiotic groups. The properties of paraprobiotics are dependent on the method of inactivation, the intensity of the method employed and on the strain used.

Bread as probiotic carriers: Resistance of Bacillus coagulans GBI-30 6086 spores through processing steps.

This study evaluated the resistance of Bacillus coagulans GBI-30, 6086 (BC) spores through the processing and storage of white and whole wheat bread. The physicochemical parameters of the probiotic bread formulations were also characterized. Loaves of bread containing or not BC were prepared. Throughout the processing, samples were collected (after mixture, after fermentation, and after baking) for enumeration of BC. In addition, BC was enumerated in different parts of loaves of bread (crust, crumb, and whole slice) collected after baking (day zero) and at different storage times (3, 7, and 10 days). The incorporation of BC did not affect the moisture, specific volume, texture and color parameters, water activity, and pH of loaves of bread. Mixing and fermentation steps did not reduce the BC survival in white or whole wheat bread. The highest (p < 0.05) number of decimal reductions (γ) was caused by baking in the crust for both loaves of bread. Baking caused around two γ of BC in the crust and 1.5 γ of BC in crumb and a whole slice of white and whole bread. Generally, storage did not increase the γ caused by baking, regardless of the evaluated part or type of bread. Results show the impacts of baking on BC and highlight the formulated white and whole wheat loaves of bread as suitable carriers for delivering the probiotic BC.

Development of a semi-dynamic in vitro model and its testing using probiotic Bacillus coagulans GBI-30, 6086 in orange juice and yogurt.

A dynamic system mimicking the gastrointestinal tract (GIT) conditions (fluids, pH, temperature, and residence time) was used to evaluate the behavior of Bacillus coagulans GBI-30, 6086 (BC) incorporated in yogurt and orange juice. BC counts were monitored in samples collected before the in vitro digestion, after initial contact with gastric fluids (30 min), static (1 h 15 min) and dynamic (2 h) stages in the gastric compartment, static (3 h) and dynamic (4 h) stages in the duodenal compartment, static (5 h) and dynamic (6 h) stages in the jejunal compartment, and after digestion. BC presented high survival in juice and yogurt over the digestion stages. The number of decimal reductions (γ) of BC caused by exposure to simulated GIT conditions was ≥0.89 in orange juice and ≥1.17 in yogurt. No differences (p ≥ 0.05) were observed on the survival of BC among the samples collected over the digestion in juice or yogurt, or between these matrices. After the in vitro digestion, BC counts were ≥7 log CFU/mL or g. Results show the great survival of BC under GIT conditions and suggest both, juice and yogurt as appropriate carries for delivering this probiotic to the diet. The semi-dynamic in vitro system was easily built and to operate, comprising an intermediate approach to assess the resistance of probiotic or potentially probiotic strains under simulated gut conditions.

Quantifying the impact of eight unit operations on the survival of eight Bacillus strains with claimed probiotic properties.

This study assessed the impact of eight unit operations [slow pasteurization, high-temperature short time (HTST) pasteurization, cooking, baking, drying, fermentation, supercritical carbon dioxide (CO2), irradiation and extrusion] in different food matrices (milk, orange juice, meatballs, bread, crystallized pineapple, yogurt, orange juice, ground black pepper, snacks, and spaghetti) on the resistance of eight (Bacillus flexus Hk1 Bacillus subtilis Bn1, Bacillus licheniformis Me1, Bacillus mojavensis KJS3, Bacillus subtilis PXN21, Bacillus subtilis PB6, Bacillus coagulans MTCC 5856 and Bacillus coagulans GBI-30, 6086) Bacillus strains with claimed probiotic properties (PB). The number of decimal reductions (γ) caused by the unit operations varied (p < 0.05) amongst the PB. Most of the unit operations caused ≤ 2 γ of PB in the food matrices evaluated. Irradiation caused up to 4.9 γ (p < 0.05) amongst the PB tested. B. subtilis Bn1, B. mojavensis KJS3, B. licheniformis Me1, and B. coagulans GBI-30 showed higher resistance to most of the tested unit operations. These results indicate that the choice of PB for application in foods should also be based on their resistance to unit operations employed during processing. Finally, the high resistance of PB to the unit operations tested comprise valuable data for the development and diversification of probiotic foods with sporeforming strains with claimed probiotic properties.

Orange Juice and Yogurt Carrying Probiotic Bacillus coagulans GBI-30 6086: Impact of Intake on Wistar Male Rats Health Parameters and Gut Bacterial Diversity.

This study aimed to investigate the impact of the food matrix (orange juice and yogurt) on the effects of the spore-forming probiotic microorganism Bacillus coagulans GBI-30 6086 in health parameters and gastrointestinal tract (gut) bacterial diversity in Wistar male rats. Rats (n = 48) were randomly distributed into six groups. The groups were the Control (which received sterile distilled water), Juice (which received orange juice), Yogurt (which received yogurt), Probiotic Bacillus (which received B. coagulans GBI-30 6086 in distilled water), Probiotic Juice (which received orange juice with B. coagulans GBI-30 6086), and Probiotic Yogurt (which received yogurt with B. coagulans GBI-30 6086). Each animal belonging to the different groups was treated for 21 days. The daily administration of probiotic juice or probiotic yogurt did not affect the rats' food or body weight. Rats fed with Probiotic Yogurt showed lower glucose and triglycerides levels (p < 0.05) in comparison to the control group (p < 0.05), while no changes in these parameters were observed in the rats fed with Probiotic Juice. Rats fed with Probiotic Yogurt showed a higher gut bacterial diversity than the control group (p < 0.05), and higher abundance (p < 0.05) of Vibrionales, Enterobacteriales, Burkholderiales, Erysipelotrichales, and Bifidobacteriales compared to all other groups. No changes were observed in the expression levels of antioxidant enzymes or heat shock protein 70 of rats fed with probiotic yogurt or probiotic juice. Results reveal that the consumption of yogurt containing B. coagulans GBI-30 6086 decreases triglycerides and glucose levels and positively impacts the gut bacterial ecology in healthy rats. These animal model findings indicate that the matrix also impacts the functionality of foods carrying spore-forming probiotics. Besides, this research indicates that yogurt is also a suitable food carrier of Bacillus coagulans GBI-30 6086.

Paraprobiotic obtained by ohmic heating added in whey-grape juice drink is effective to control postprandial glycemia in healthy adults.

The effect of paraprobiotic Lacticaseibacillus casei 01 inactivated by ohmic heating (8 V/cm, 95 °C/7 min, 60 Hz) whey-grape juice drink at the postprandial glycemia was evaluated. In vitro hypoglycemic activity was assessed by the α-glucosidase and α-amylase inhibition, while in vivo activity was determined using 15 healthy subjects, which consumed bread + probiotic whey drink, bread + paraprobiotic whey drink, and bread alone as a control. The probiotic and paraprobiotic grape-flavored whey drinks showed similar α-glucosidase and α-amylase inhibition (51.2 vs 51.8% and 43.2 vs 44.2%, respectively). The consumption of both paraprobiotic and probiotic whey drinks increased the incremental glucose rate when compared to the control due to the presence of sugar in its composition, without changes in the other parameters evaluated (maximum glucose value, glucose incremental percentage, and peak blood glucose time), showing a reduced glycemic response. In addition, the consumption of the paraprobiotic drink maintained the maximum glucose increase similar to the control, while an increase in this parameter was observed after the consumption of the probiotic drink. Therefore, the paraprobiotic grape-flavored whey drink may be an effective alternative to replace the probiotic product in reducing the postprandial glycemia in healthy individuals.

Ohmic heating as a method of obtaining paraprobiotics: Impacts on cell structure and viability by flow cytometry.

This study aimed to evaluate the effects of ohmic heating (OH) on probiotic inactivation, cell viability and morphology of the probiotic strains Lactobacillus acidophilus LA 05 (LA), Lacticaseibacillus casei 01 (LC), and Bifidobacterium animalis Bb 12 (BA) to develop paraprobiotics. OH at different electric field magnitudes (4, 8, and 12 V/cm at 60 Hz) and conventional heat treatment (CONV) were performed to determine the most adequate condition for the obtainment of paraprobiotics. Analysis of culturability, flow cytometry (FC), and Scanning electron microscope (SEM) was carried out. The complete inactivation by CONV was achieved only in the following conditions: LA - 95 °C/5 min, LC and BA - 95 °C/7 min. The same temperature profile was used in OH treatments to study the OH electrical effects. The OH treatment (8 V/cm) caused lower damage to the cell membrane integrity compared to the CONV treatment (p < 0.05). The OH showed to be adequate technology for the efficient production of paraprobiotics.

Behavior of different Bacillus strains with claimed probiotic properties throughout processed cheese ("requeijão cremoso") manufacturing and storage.

Even though spore-forming bacteria have been mainly linked to spoilage or foodborne pathogens vehiculated through foods, some strains of Bacillus can potentially present probiotic properties. The advantage of incorporating probiotic Bacillus strains in foods relies mainly on the fact that these microorganisms present high resistance to harsh processing conditions. "Requeijão cremoso" is a type of processed cheese highly appreciated in Brazil. During processing, this product is submitted to several harsh conditions (heating at 90 °C, for instance), leading to the inactivation of probiotic bacteria belonging to Lactobacillus and Bifidobacterium genera. That fact has precluded the development of probiotic "requeijão cremoso" products; however, probiotic Bacillus strains may comprise a promising alternative to overcome the low resistance of traditional probiotics to food processing. The objective of this study was to evaluate the behavior of different Bacillus strains with claimed probiotic properties throughout processed cheese ("requeijão cremoso") manufacturing. A total of five different Bacillus strains with claimed probiotic properties (B. coagulans MTCC 5856, B. coagulans GBI-30 6086, B. subtilis PXN 21, B. subtilis PB6, and B. flexus HK1) were individually inoculated at different stages of manufacture - curd pasteurization, coagulation, and fusion - of "requeijão cremoso" and their survival in each of these stages was determined. The survival of B. coagulans GBI-30 6086 was further assessed throughout "requeijão cremoso" production and shelf life (45 days at 6 °C). Besides, the chemical composition, level of proteolysis, and fatty acid profile of the treatments during shelf life were evaluated. The fusion stage was found as the most appropriate for the addition of B. coagulans GBI-30 6086, which allows the production of probiotic "requeijão cremoso" and facilitates the technological process while preventing the occurrence of final product recontamination.