A Big World in Small Grain: A Review of Natural Milk Kefir Starters

Milk kefir is a traditional fermented milk product whose consumption is becoming increasingly popular. The natural starter for kefir production is kefir grain, which consists of various bacterial and yeast species. At the industrial scale, however, kefir grains are rarely used due to their slow growth, complex application, bad reproducibility and high costs. Instead, mixtures of defined lactic acid bacteria and sometimes yeasts are applied, which alter sensory and functional properties compared to natural grain-based milk kefir. In order to be able to mimic natural starter cultures for authentic kefir production, it is a prerequisite to gain deep knowledge about the nature of kefir grains, its microbial composition, morphologic structure, composition of strains on grains and the impact of environmental parameters on kefir grain characteristics. In addition, it is very important to deeply investigate the numerous multi-dimensional interactions among different species, which play important roles on the formation and the functionality of grains

Development of Antioxidant Activity during Milk Fermentation by Wild Isolates of Lactobacillus helveticus

Background and Objective: Oxidative stress, due to free radicals, brings injury to the body by attacking large molecules and cell organs, and is the main reason of many diseases. Fermentation of foods containing large amount of proteins such as milk by special species of lactic acid bacteria is a potential way in enhancement of the antioxidative activity of foods. This study aimed at evaluating non-common starter species isolates of Lactobacillus helveticus for their capability to produce fermented milk enriched in antioxidant peptides.Materials and Methods: Reconstituted skim milk (11%) was inoculated with 7 wild isolates of Lactobacillus helveticus, and after 24 h fermentation at 37ºC, the samples were kept 4ºC and for 14 days. Viable cell number, acidification and proteolysis degree in the milk fermented by each isolate were assessed in 1, 7 and 14 days. Development of antioxidant activity was measured using DPPH and ABTS●+ radial scavenging activities during the storage period.Results and Conclusion: Though some slight strain-dependent differences were observed in growth, acidification and proteolysis, all the samples showed considerably strong antioxidant activity (at least 62.32±3.66% and 57.64±1.42% measured using DPPH and ABTS●+ radicals, respectively) through the whole storage period. In vitro simulated gastrointestinal digestion indicated that DPPH radical-scavenging activity of the antioxidative peptidic supernatants was not affected significantly by consecutive pepsin-pancreatin hydrolysis in most of the samples. These evidences support Lactobacillus helveticus as a promising functional culture able to promote health benefits in dairy-based functional foods.Conflict of interest: The authors declare that there is no conflict of interest

In Vitro characterization of Lactococcus lactis strains Isolated from Iranian Traditional Dairy Products as a Potential Probiotic

Few studies have been reported regarding probiotic properties of Lactococcus lactis strains although they are extensively used as starter cultures in the production of dairy products. In this study 8 wild isolates of Lactococcus lactis were evaluated in vitro with regard to resistance to simulated gastric and intestinal juices, adherence ability to Caco-2 cells and HT29-MTX-E12 cell lines, anti-microbial activity, hydrophobicity and antibiotic susceptibility. The results revealed that all isolates had better survival after exposure to simulated gastrointestinal tract stresses in comparison to control probiotic Lactobacillus rhamnosus GG. Regarding adherence efficiency, almost all isolates exhibited similar adherence with control. Three isolates showed antibacterial activity against Gram-positive pathogens (Staphylococcus aureus and Listeria monocytogenes) through spot-agar method. Almost all isolates (seven out of eight) showed similar hydrophobicity to control probiotic. Regarding to antibiotic resistance, all isolates were susceptible to gentamicin, ampicillin, ciprofloxacin, erythromycin, tetracycline, penicillin, kanamycin and nitrofurantoin. Although, further investigations are necessary, it was concluded that strains derived from raw milk and home-made dairy products could be a remarkable reservoir for identification of new potential probiotic strains

Screening of Lactic Acid Bacteria Isolated from Iranian sourdoughs for Antifungal Activity: Enterococcus faecium showed the Most Potent Antifungal Activity in Bread

Background and Objective: The use of antifungal lactic acid bacteria as starter for bread making could be a good alternative to improve the stability of bread shelf life.Material and Methods: In this study, a total of 57 lactic acid bacteria were isolated from spontaneously fermented wheat sourdoughs collected in Chahar-Mahalo Bakhryari province of Iran. The isolates were screened for in vitro antifungal activity (towards Aspergilus niger or Penicillium roqueforti); and the selected isolates (six isolates) were applied in flat bread making. The freshly baked breads were nebulized with a suspension of either molds, containing 104 spores ml-1, and the fungal growth on breads was monitored over a 7-day storage period.Results and Conclusion: Bread produced with either isolates AN3 and MB1 (both were identified as Enterococcus faecium) restrained the growth of Aspergillus niger for up to 5 days. Even though none of the isolates were strong enough to inhibit the growth of Penicillium roquforti on bread, the surface area of breads contaminated by this fungus was significantly lower than the control samples. To our knowledge, it was the first report indicating the anti-mold activity of Enterococcus faecium strains isolated from sourdough. These isolates seem to be promising for further analysis and their application in bread industry for prolonging the shelf life.Conflict of interest: The authors declare that there is no conflict of interest

Manufacture of a functional fermented milk enriched of Angiotensin-I Converting Enzyme (ACE)-inhibitory peptides and γ-amino butyric acid (GABA)

Abstract This study aimed at developing and characterizing a fermented milk with a potentially anti-hypertensive effect due to the concurrent presence of Angiotensin-I Converting Enzyme (ACE)-inhibitory peptides and γ-amino butyric acid (GABA). Preliminarily, lactic acid bacteria strains were screened based on the capacity of releasing ACE-inhibitory peptides and of synthesizing GABA. The most potent RP-FPLC fraction from the milk fermented with Lactococcus lactis DIBCA2 had an ACE-inhibitory activity, expressed in terms of IC 50 , of 5 ± 2 μg/mL, a value which compares well to those reported for the majority of ACE-inhibitory peptides identified in fermented milks. This fraction contained a mixture of six peptides, five of which shared motifs in common with ACE-inhibitory peptides previously reported. Lactobacillus plantarum PU11 was selected as the highest producer of GABA, ca. 77.4 mg/kg after 120 h of milk fermentation. Both the strains were selected and used as starters for consecutive milk fermentation. After fermentation, the milk contained both the lactic acid bacteria at the cell density of ca. 8.0 log CFU/mL, had a pH of ca. 4.45, and showed ACE-inhibitory activity (IC 50 = 0.70 ± 0.07 mg/mL) and a concentration of GABA (ca. 144.5 mg/kg), which were compatible with the dosage for mild antihypertensive effect

Quantification of Major Bacteria and Yeast Species in Kefir Consortia by Multiplex TaqMan qPCR

Kefir grains are complex microbial systems of several groups of microorganisms. The identification and quantification of the microbial composition of milk kefirs was described in several studies, which provided an insight into the microbial consortia in this complex ecosystem. Nevertheless, the current methods for identification and quantification are not appropriate for deeper studies on kefir consortia, e.g., population dynamics and microbial interactions in kefir grains. This requires another sensitive and reliable quantitative method. Therefore, this study aims to develop multiplexed qPCR assays to specifically detect and quantify, as an example, several microorganisms of the milk kefir microbial community. Primer-probe sets, which target species-specific genes in six bacteria and five yeasts, were designed, and their sensitivity and specificity to the target species was analyzed in simplex as well as four multiplex qPCR assays. The self-designed multiplex assays were applied for the detection of target bacteria and yeast species in milk kefirs, in both, grain and beverage fractions. Detection of all target microorganisms in simplex and multiplex qPCR was achieved by good linearity, efficiency, repeatability and reproducibility in all assays. When the designed assays were applied on six kefirs, all target microorganisms were detected in different samples, but not all in one kefir sample. The two ubiquitous lactobacilli Lactobacillus kefiranofaciens and Lb. kefiri were present in all six kefirs studied, but were associated with different other yeasts and bacteria. Especially on the yeast community a significant diversity was observed. In general, multiplex TaqMan qPCR as developed here was proven to have high potential for specific identification of target microorganisms in kefir samples and for the first time, eleven target bacteria and yeasts of kefir microbiota were rapidly detected and quantified. This study, thus, provides a fast and reliable protocol for future studies on kefir and other similar microbial ecosystems.DFG, 414044773, Open Access Publizieren 2019 - 2020 / Technische Universität Berli

The Efficiency of Health Performance in Universities of Medical Sciences in Iran: Data Envelopment Analysis

Background: Technical efficiency, which is measured by calculating the ratio of products to resources, is the most important factor in assessing the efficiency status of organizations. Data envelopment analysis is useful to measure the efficiency score of all the units which have homogeneous input resources and output products and to rank them. The aim of this study was to measure and compare the efficiency of health performance in medical universities in Iran.Methods: The present research is a cross-sectional study to measure the efficiency of health performance using the national information of the health system of Iran. Input data include hospital beds, specialists, general physicians, dentists, pharmacists, nurses, midwives, computerised topography scan and magnetic resonance imagination devices, and Gini Index; also, the output data include pregnancy care coverage, infant mortality rate, low birth weight, and in-patient days. These data were attained from the annual Ministry of Health and Medical Education report in 2017 for 46 medical universities. To estimate the efficiency of health performance of each medical university using data envelopment analysis, we designed an input-oriented model with Variable Returns to Scale in GAMS 28.2.0. The effect of contextual factors on the efficiency score was calculated using the Tobit Regression model.Results: Results showed that only 19 (41%) medical universities were on the efficiency frontier. The highest mean of efficiency score was attributed to eastern areas, followed by the western and northern areas, and the worst status was related to southern parts of the country. The efficiency scores of universities located in northern areas were closer, while there was more difference among the efficiency scores of the universities of central areas of the country. Tobit regression shows that significant factors inefficiency include life expectancy and medical university class.Conclusion: The results of this study emphasized the differences in the performance efficiency of medical universities. Considering the inefficiency of smaller universities, we need to make careful decisions in establishing new universities in small cities

An evaluation of the effect of pulsed wave low-level laser therapy on the biomechanical properties of the vertebral body in two experimental osteoporosis rat models.

Osteoporosis (OP) increases vertebral fragility as a result of the biomechanical effects of diminished bone structure and composition. This study has aimed to assess the effects of pulsed wave low-level laser therapy (PW LLLT) on cancellous bone strength of an ovariectomized (OVX-d) experimental rat model and a glucocorticoid-induced OP (GIOP) experimental rat model. There were four OVX-d groups and four dexamethasone-treated groups. A group of healthy rats was used for baseline evaluations. The OVX-d rats were further subdivided into the following groups: control rats with OP, OVX-d rats that received alendronate, OVX-d rats treated with PW LLLT, and OVX-d rats treated with alendronate and PW LLLT. The remaining rats received dexamethasone and were divided into four groups: control, alendronate-treated rats, laser-treated rats, and laser-treated rats with concomitant administration of alendronate. PW LLLT (890 nm, 80 Hz, 0.972 J/cm(2)) was performed on the spinal processes of the T12, L1, L2, and L3 vertebras. We extracted the L1 vertebrae and submitted them to a mechanical compression test. Biomechanical test findings showed positive effects of the PW LLLT and alendronate administration on increasing bending stiffness and maximum force of the osteoporotic bones compared to the healthy group. However, laser treatment of OVA-d rats significantly increased stress high load compared to OVA-d control rats. PW LLLT preserved the cancellous (trabecular) bone of vertebra against the detrimental effects of OV-induced OP on bone strength in rats compared to control OV rats

Volatilomics-Based Microbiome Evaluation of Fermented Dairy by Prototypic Headspace-Gas Chromatography–High-Temperature Ion Mobility Spectrometry (HS-GC-HTIMS) and Non-Negative Matrix Factorization (NNMF)

Fermented foods, such as yogurt and kefir, contain a versatile spectrum of volatile organic compounds (VOCs), including ethanol, acetic acid, ethyl acetate, and diacetyl. To overcome the challenge of overlapping peaks regarding these key compounds, the drift tube temperature was raised in a prototypic high-temperature ion mobility spectrometer (HTIMS). This HS-GC-HTIMS was used for the volatilomic profiling of 33 traditional kefir, 13 commercial kefir, and 15 commercial yogurt samples. Pattern recognition techniques, including principal component analysis (PCA) and NNMF, in combination with non-targeted screening, revealed distinct differences between traditional and commercial kefir while showing strong similarities between commercial kefir and yogurt. Classification of fermented dairy samples into commercial yogurt, commercial kefir, traditional mild kefir, and traditional tangy kefir was also possible for both PCA- and NNMF-based models, obtaining cross-validation (CV) error rates of 0% for PCA-LDA, PCA-kNN (k = 5), and NNMF-kNN (k = 5) and 3.3% for PCA-SVM and NNMF-LDA. Through back projection of NNMF loadings, characteristic substances were identified, indicating a mild flavor composition of commercial samples, with high concentrations of buttery-flavored diacetyl. In contrast, traditional kefir showed a diverse VOC profile with high amounts of flavorful alcohols (including ethanol and methyl-1-butanol), esters (including ethyl acetate and 3-methylbutyl acetate), and aldehydes. For validation of the results and deeper understanding, qPCR sequencing was used to evaluate the microbial consortia, confirming the microbial associations between commercial kefir and commercial yogurt and reinforcing the differences between traditional and commercial kefir. The diverse flavor profile of traditional kefir primarily results from the yeast consortium, while commercial kefir and yogurt is primarily, but not exclusively, produced through bacterial fermentation. The flavor profile of fermented dairy products may be used to directly evaluate the microbial consortium using HS-GC-HTIMS analysis

Transcriptional drug repositioning and cheminformatics approach for differentiation therapy of leukaemia cells.

Differentiation therapy is attracting increasing interest in cancer as it can be more specific than conventional chemotherapy approaches, and it has offered new treatment options for some cancer types, such as treating acute promyelocytic leukaemia (APL) by retinoic acid. However, there is a pressing need to identify additional molecules which act in this way, both in leukaemia and other cancer types. In this work, we hence developed a novel transcriptional drug repositioning approach, based on both bioinformatics and cheminformatics components, that enables selecting such compounds in a more informed manner. We have validated the approach for leukaemia cells, and retrospectively retinoic acid was successfully identified using our method. Prospectively, the anti-parasitic compound fenbendazole was tested in leukaemia cells, and we were able to show that it can induce the differentiation of leukaemia cells to granulocytes in low concentrations of 0.1 μM and within as short a time period as 3 days. This work hence provides a systematic and validated approach for identifying small molecules for differentiation therapy in cancer