Draft genome sequence of Lactobacillus helveticus OSU-BDGOAK2 and Lactobacillus kefiranofaciens OSU-BDGOA1, kefir grain isolates with potential antibacterial activity.

We present the draft genome sequence and assembly of Lactobacillus helveticus OSU-BDGOAK2 and Lactobacillus kefiranofaciens OSU-BDGOA1 isolated from kefir grains that exhibited in vitro antibacterial activity against Escherichia coli ATCC 25922, Listeria innocua ATCC 51742, and Staphylococcus epidermidis ATCC 1222. Genome analysis of both strains revealed gene clusters encoding bacteriocins.

Enhanced probiotic potential of Lactobacillus kefiranofaciens OSU-BDGOA1 through co-culture with Kluyveromyces marxianus bdgo-ym6.

Introduction: Due to the increasing consumer demand for the development and improvement of functional foods containing probiotics, new probiotic candidates need to be explored as well as novel means to enhance their beneficial effects. Lactobacillus kefiranofaciens OSU-BDGOA1 is a strain isolated from kefir grains that has demonstrated probiotic traits. This species is the main inhabitant of kefir grains and is responsible for the production of an exopolysaccharide (EPS) whit vast technological applications and potential bioactivities. Research has shown that interkingdom interactions of yeast and lactic acid bacteria can enhance metabolic activities and promote resistance to environmental stressors. Methods: Comparative genomic analyses were performed to distinguish OSU-BDGOA1 from other strains of the same species, and the genome was mined to provide molecular evidence for relevant probiotic properties. We further assessed the cumulative effect on the probiotic properties of OSU-BDGOA1 and Kluyveromyces marxianus bdgo-ym6 yeast co-culture compared to monocultures. Results: Survival during simulated digestion assessed by the INFOGEST digestion model showed higher survival of OSU-BDGOA1 and bdgo-ym6 in co-culture. The adhesion to intestinal cells assessed with the Caco-2 intestinal cell model revealed enhanced adhesion of OSU-BDGOA1 in co-culture. The observed increase in survival during digestion could be associated with the increased production of EPS during the late exponential and early stationary phases of co-culture that, by enhancing co-aggregation between the yeast and the bacterium, protects the microorganisms from severe gastrointestinal conditions as observed by SEM images. Immune modulation and barrier function for recovery and prevention of flagellin-mediated inflammation by Salmonella Typhimurium heat-killed cells (HKSC) in Caco-2 cells were also measured. OSU-BDGOA1 in mono- and co-culture regulated inflammation through downregulation of pro-inflammatory cytokine expression and increased membrane barrier integrity assessed by TEER, FD4 permeability, and expression of tight junctions. Discussion: The results of the study warrant further research into the application of co-cultures of yeast and LAB in functional probiotic products and the potential to increase EPS production by co-culture strategies.

Mitigation of undesirable volatile aroma compounds in kefir by freeze drying and vacuum evaporation.

Commercial kefir was recently found to be effective in curing recurrent Clostridioides difficile infection when consumed alongside antibiotic treatment. However, kefir products have limited acceptance among Western consumers due to their characteristic flavor and texture. Plain, unsweetened commercial kefir with 1% milkfat was subjected to vacuum evaporation and freeze-drying processes to assess the effect on volatile organic compound concentration, sensory quality, and microbial viability. Vacuum evaporation and freeze-drying both significantly decreased the concentration of 26 out of 27 volatiles in the kefir (p < 0.05), by an average of 61%. Vacuum evaporation exhibited an apparent greater reduction than freeze-drying in the concentrations of the ketones and aldehydes, while freeze-drying showed a more noticeable reduction in the concentrations of the acids, alcohols, and esters. While lowering the concentration of volatiles did not significantly improve the average consumer acceptance of commercial kefir, both treatments showed differences in the range of acceptability responses. Both treatments also produced a small but significant reduction in the concentrations of Lactobacillus and Lactococcus species present.

Metagenomic analysis and antibacterial activity of kefir microorganisms.

The microbiota composition of kefir grain and milk kefir was assessed via a metagenomic approach. Significant microorganisms were isolated and identified using molecular methods. A safety assessment was conducted based on antibiotic susceptibility and blood hemolysis. Probiotic traits such as resistance to gastric tract conditions, surface characteristics, adhesion to intestinal cells, and antibacterial activity were also assessed. Metagenomic analysis revealed that kefir grains are a more stable community with clear dominant species as compared to milk kefir. Lactobacillus kefiranofaciens BDGO-A1, Lactobacillus helveticus BDGO-AK2, and Lactobacillu kefiri strains showed tolerance to acidic pH and the presence of bile salts, adhesion capability to Caco-2 cells, in vitro antibacterial activity, and the production of antibacterial proteins. In the metagenomic analysis, contigs associated with these species showed the presence of genes involved in exporting polyketide antibiotics and bacteriocin production. To fully exploit the potential probiotic properties of these microorganisms to help human health, further investigation is necessary to elucidate the mechanisms behind the biological activity and the genotypic characteristics of the isolated strains.

Invited review: Milk kefir microbiota-Direct and indirect antimicrobial effects.

Kefir is a fermented dairy product with well recognized probiotic properties. Recently, consumer interest in fermented products with probiotic microorganisms has increased due to the accumulating evidence of the effects of kefir microorganisms on the modulation of gut microbiota and their antimicrobial activity. Although the health properties of kefir have been reviewed in other works, the present review addresses the antimicrobial effects of kefir microbiota and associated compounds. The antimicrobial activity of kefir microorganisms could derive from different mechanisms. The microorganisms' capacity to adhere to the intestinal epithelium, preventing the adhesion of pathogens, and their immunomodulation properties are among the mechanisms suggested. Bacteria and yeast isolated from kefir have been shown to have in vivo and in vitro antimicrobial activity against enteropathogenic bacteria and spoilage fungi. However, most reports have focused their approach on single-strain antimicrobial properties; evaluation of antimicrobial activity of cocultures of kefir microbiota and their potential mechanisms of action has been neglected. Kefir microbiota and associated compounds have shown promising antimicrobial effects; however, more research needs to be done to discern the mechanisms of action.

Functional Properties of a High Protein Beverage Stabilized with Oat-ß-Glucan.

This study evaluated the effect of oat flour and milk protein on the functional properties and sensory acceptability of shelf stable high protein dairy beverages containing at least 0.75 g of oat-ß-glucan per serving size. Formulations adjusted to levels of 1.50% to 2.30% oat flour and 2.50% to 4.00% milk protein isolate (MPI) were thermal processed in a rotary retort. The finished product exhibited good suspension stability (>80%). The increase of oat and MPI contents lead to nectar-like beverages (51 to 100 mPas). However, oat flour was the component showing the highest effect on the viscosity coefficient values of the beverages. Sensory evaluation indicated that formulations with less than 1.9% oat flour and 2.5% MPI (thin liquid, <50 mPas) were the most accepted. Mouthfeel (perceived thickness), sweetness and aftertaste had the most influence on overall liking of the beverages. PRACTICAL APPLICATION: Overall, this study comprises the development of a functional food product. Supplementation of beverages with fiber from oats is an innovative approach to stabilize high protein beverages. Ready to drink protein beverage formulations use gums to stabilize the product and provide a desirable mouthfeel. The levels of oat-ß-glucan used in the beverage increased the thickness and meet the requirement of the FDA approved health claim for reduction of the cardiovascular disease risk (21 CFR 101.81).

Antihyperlipidemic effect of methanolic extract from Opuntia joconostle seeds in mice fed a hypercholesterolemic diet.

The purpose of this study was to evaluate the hypolipidemic effect of a methanolic extract from Opuntia joconostle seeds fed to mice in a hypercholesterolemic diet. Acute toxicity of the methanolic extract was investigated by an established method. Phenolic composition and antioxidant activity were determined by high-performance liquid chromatography and DPPH, respectively. The total phenolic content of Opuntia joconostle seeds was 47.85 ± 1.29 mg gallic acid equivalents/g dry weight. The main phenolic compounds were identified as quercetin, rutin, and cafeic acid. Percent inhibition of DPPH⁺ was 49.76 ± 0.49 %. The oral LD50 for the methanolic extract from the Opuntia joconostle seeds was >5,000 mg/kg BW. Mice fed a hypercholesterolemic diet for six days exhibited significantly (P ≤ 0.001) higher plasma lipid levels than mice fed a normal diet. Remarkably, supplementation with methanolic extract from Opuntia joconostle at doses of 1, 2, and 5 g/kg body weight significantly (P ≤ 0.001) prevented the increase in total cholesterol, low-density lipoprotein cholesterol, triglycerides level, and atherogenic index. Similar concentrations of the HDL cholesterol were observed in both treated and control groups. A significant dose-dependent reduction in lipid levels was noted for treated groups compared to the hypercholesterolemic group. We attribute this result to the seeds' phenolic composition. This methanolic extract has potential to be included in short-term hypercholesterolemia treatment regimens as it exhibits hypolipidemic activity with no apparent toxic manifestations.

Ohmic heating of peaches in the wide range of frequencies (50 Hz to 1 MHz).

UNLABELLED: The ohmic heating (OH) rate of peaches was studied at fixed electric field strength of 60 V.cm⁻¹, square-shaped instant reversal bipolar pulses, and frequencies varying within 50 Hz to 1 MHz. Thermal damage of tissue was evaluated from electrical admittivity. It showed that the time for half disruption (τ(T)) of tissue was required more than 10 h at temperatures below 40 °C. However, cellular thermal disruption occurred almost instantly (τ(T) < 1 s) at high temperatures (> 90 °C). Electrical conductivity σ(o) and admittivity σ(o)* of tissue at T(o)= 0 °C and their temperature coefficients (m, m*) were calculated. For freeze-thawed tissues, σ and σ* as well as m and m* were nearly indifferent to the frequency. However, for the intact tissue, both σ(o), σ(o)* and m, m* were frequency dependent. For freeze-thawed product, the power factor (P) was approximately equal to 1 and indifferent to the frequency and temperature. On the other hand, strong frequency dependence was observed for intact tissue with the minimum P approximately equal to 0.68 in the range of tens of kHz. The time required to reach a target temperature t(f) was evaluated. The t(f) increased with frequency up to the middle of the range of tens of kHz and thereafter continuously decreased. Samples exposed to the low-frequency electric field demonstrated faster electro-thermal damage rates. The textural relaxation data supported more intense damage kinetics at low-frequency OH. It has been demonstrated that a combination of high-frequency OH with pasteurization at moderate temperature followed by rapid cooling minimizes texture degradation of peach tissue. PRACTICAL APPLICATION: In this study, we investigated the electric field frequency effect on the rate of OH of peaches. It was shown that the time required for reaching the target temperature is strongly dependent upon the frequency. Samples exposed to low-frequency OH demonstrated higher electro-thermal damage rates. It has been shown that the combination of high-frequency OH with pasteurization at moderate temperature followed by rapid cooling minimizes texture degradation of peach tissue. Obtained results provide new information on the impact of electric field frequency on OH, which is useful for OH process design.