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Adhesion of Lactobacillus strains on Caco2 cell cultures observed under oil immersion microscope (100X) after staining with Geimsa strain. A-Blank Caco2 cell line, B- L. plantarumCSCC 5276, C- L. plantarumLp91, D- L. plantarumLp9, E- L. plantarumLp72, F- L. plantarumLp75, G- L. plantarumLp77, H- L. plantarumLp90, I- L. delbrueckiisubsp. bulgaricusCH4.
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Adherence of bacteria to epithelial cells and mucosal surfaces is a key criterion for selection of probiotic. We assessed the adhesion property of selected indigenous probiotic Lactobacillus strains based on their hydrophobicity and ability to adhere to human epithelial cells.
Five human faecal Lactobacillus isolates, one from buffalo milk and one...
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Citations
... All the tested strains, L. fermentum RCEID47-7, L. fermentum RCEID 23-2, and L. oris RCEID 28-3, had similar abilities to adhere to both cell lines, and the highest adhesion level was obtained for the RCEID47-7 strain. Our results are in concordance with those of a previous study that demonstrated that the selected probiotic had similar adhesion capabilities for both Caco-2 and HT-29 cell lines (Duary et al., 2011). Factors affecting the adhesion of human intestinal mucosa by probiotic bacteria are mediated by a protein called "adhesin" on the bacterial cell surface to the mucus or intestinal epithelial cells (van Pijkeren et al., 2006). ...
Several bacterial genera, including Lactobacillus, have been evaluated for their beneficial effects in humans. This study aimed to evaluate the “probiotic” and “postbiotic” potential of three bacterial strains: Lactobacillus oris RCEID28-3, Limosilactobacillus fermentum RCEID23-2, and Limosilactobacillus fermentum RCEID47-7. Probiotics are defined as live bacterial cells, whereas postbiotics are inactivated or killed probiotics. Probiotic potential was evaluated by assessing the adhesion ability of the Caco-2 and HT-29 cell lines, immunomodulatory effects, and cholesterol assimilation. Meanwhile, postbiotic potential was assessed by evaluating their immunomodulatory effects through measuring cytokine production in human peripheral blood mononuclear cells (PBMCs) using an enzyme-linked immunosorbent assay (ELISA). The human intestinal cell adhesion assay showed that all bacterial strains exhibited adhesion of more than 90% to both the Caco-2 and HT-29 cell lines, except for L. oris RCEID28-3. The highest cell adhesion level was observed in L. fermentum RCEID47-7. The cytokine production assay revealed that probiotic and postbiotic strains stimulated the production of three cytokines: IL-10, IFN- γ, and TNF-α, in PBMCs with the different cytokines, indicating the strain-dependent property. Moreover, cholesterol assimilation by live probiotics showed that all the strains, especially L. fermentum RCEID47-7, could reduce cholesterol levels. Therefore, this study provides scientific evidence to support the possibility of applying probiotics and their inactivated forms (postbiotics) in humans in the near future.
... The binding potential of probiotics to intestinal epithelial cells is another selective characteristic of probiotics. Therefore, adherence ability is considered a critical criterion for selecting potential probiotic bacteria 51 . According to the results, strains Y33 (48.50%) and Y17 (26.40%) showed the strongest adhesion to human intestinal CaCO2 cells. ...
Today, probiotics are considered to be living microorganisms whose consumption has a certain number of beneficial effects on the consumer. The present study aimed to investigate the effect of a new probiotic extract (Lactobacillus delbrueckii subsp. lactis KUMS Y33) on the differentiation process of human adipose-derived stem cells (hADSCs) into adipocytes and osteocytes and, as a result, clarify its role in the prevention and treatment of bone age disease. Several bacteria were isolated from traditional yogurt. They were evaluated to characterize the probiotic’s activity. Then, the isolated hADSCs were treated with the probiotic extract, and then osteogenesis and adipogenesis were induced. To evaluate the differentiation process, oil red O and alizarin red staining, a triglyceride content assay, an alkaline phosphatase (ALP) activity assay, as well as real-time PCR and western blot analysis of osteocyte- and adipocyte-specific genes, were performed. Ultimately, the new strain was sequenced and registered on NBCI. In the probiotic-treated group, the triglyceride content and the gene expression and protein levels of C/EBP-α and PPAR-γ2 (adipocyte-specific markers) were significantly decreased compared to the control group (P < 0.05), indicating an inhibited adipogenesis process. Furthermore, the probiotic extract caused a significant increase in the ALP activity, the expression levels of RUNX2 and osteocalcin, and the protein levels of collagen I and FGF-23 (osteocyte-specific markers) in comparison to the control group (P < 0.05), indicating an enhanced osteogenesis process. According to the results of the present study, the probiotic extract inhibits adipogenesis and significantly increases osteogenesis, suggesting a positive role in the prevention and treatment of osteoporosis and opening a new aspect for future in-vivo study.
... The microbial adhesion to hydrocarbon assay to determine the cell surface hydrophobicity of isolates and standard culture using hexadecane (Sigma, India), xylene (Sigma, India) and toluene (Sigma, India) as solvents was done. 22 Cell surface hydrophobicity in terms of percent (H%) was calculated using the following formula: ...
... 34 Duary et al. determined the cell surface hydrophobicity of ve Lactobacillus strains isolated from human faecal samples; however, the values were comparatively higher than those reported in the present study. 22 The L. rhamnosus strain showed sensitivity against all the tested antibiotics. The formation of zone of inhibitions of 13 ± 1.414, 23 ± 1.514, 17 ± 1.614, 20 ± 1.414, 24 ± 1.214, 21 ± 1.414, 27 ± 1.494, 30 ± 1.414, 12 ± 1.464, 14 ± 1.414 and 15 ± 1.514 mm against ampicillin (10 mcg), chloramphenicol (25 mcg), streptomycin (10 mcg), sulphatriad (300 mcg), tetracycline (25 mcg), bacitracin (10 units), chloramphenicol (30 mcg), penicillin G (10 units), polymyxin B (300 mcg), gentamycin (10 mcg) and neomycin (30 mcg), respectively, shows the sensitivity to different antibiotics. ...
Water chestnut (Trapa bispinosa) starch has technical and functional qualities that are mostly comparable to those of traditional sources. In this study, low-fat flavoured probiotic yogurt was prepared with the addition of Lacticaseibacillus rhamnosus (ATCC 7469), mango pulp and water chestnut starch as a fat replacer as well as a biothickener. A central composite face centered experimental design (CCFD) optimization study was carried out using different combinations of fat (0.5–1.5%), starch (1.0–3.0%), fructo-oligosaccharides (0.5–1.5%) and inulin (0.5–1.5%). L. rhamnosus exhibited resistance to simulated gastric fluid, intestinal fluid and hydrogen peroxide, along with good cell surface hydrophobicity and antimicrobial properties against Escherichia coli, Bacillus subtilis, Salmonella typhimurium and Staphylococcus aureus. Water chestnut starch was found to have a characteristic A-type crystalline pattern with 28% crystallinity, and it was smooth with oval to irregular in shape. The proximate, texture, color, and sensory properties and the viability of L. rhamnosus in yogurt did not vary significantly during a 15 day storage period. Thus, the starch from water chestnuts can be explored as a possible fat substitute for formulating low-calorie food products.
... bacteria that are resistant to low acid environment and bile salt have a higher capacity to adhere to the intestinal epithelium and mucosa(Bao et al. 2010;Guo et al. 2015;Jin et al. 1998;Ross et al. 2005). L. plantarum probiotic bacteria strains, which is known to have high adhesion ability(Duary et al. 2011), showed average adhesion capacity compared to the L. rhamnosus GG control strain in our adhesion tests. When the ndings of the L. rhamnosus species obtained from three different sources were evaluated, the probiotic isolated from adult feces showed higher adhesion capacity than the other sources. ...
Backgorund
In recent years, there has been an increasing interest in the field of research into the isolation and characterization of probiotics in the prevention of diseases and the need to maintain the continuity of healthy microbiota. Therefore, the aim of this study is to isolate and identify bacteria found in maternal colostrum, breast milk, adult and infant feces, analyze possible probiotic potential, and reveal the developmental kinetics of selected strains.
Results
We isolated 40 bacterial species from 4 different sources and identified 19 bacteria in the form of bacilli through molecular biology and carried out studies with 11 of them. 5 of the selected strains were showed the better results considering bile salt resistance and ability to survive at different pH, antimicrobial effect. When the adhesion capacity in cell culture was compared, Lactobacillus pontis ZZ6780 and Lactobacillus reuteri NBC2680 came into prominence. Furthermore, the growth kinetics of these strains were demonstrated on a 3 L bioreactor scale. Finally, the growth kinetics of selected strains were determined and the maximum specific growth rate of selected Lactobacillus pontis ZZ6780 and Lactobacillus reuteri NBC2680 was calculated as 0.412 h⁻¹ and 0.481 h⁻¹, respectively. In addition, the dry cell matter amounts were found to be and 4.45 g/L and 5.23 g/L, respectively.
Conclusion
This study established the groundwork for the selection of safety probiotics for the development and application of LAB. It is thought that the two strains obtained as a result of this study can be considered as potential probiotic strains in the food, pharmaceutical and cosmetic industries.
... Probiotics exhibit adhesion properties that contribute to a competitive exclusion effect, preventing intestinal pathogens from attaching to intestinal cells [53]. This adhesion relies on the physical interactions of van der Waals forces between surfaces [54]. Bacterial capsular polysaccharides, teichoic/lipoteichoic acids, lipoproteins, and surface proteins like mucinbinding/fibronectin-binding/collagen-binding proteins collaborate to form multi-cellular aggregates [55]. ...
Although global life expectancy has increased over the past 20 years due to improvements in managing infectious diseases, one in five people still succumbs to infections. Humanity is tirelessly working towards developing vaccines and antimicrobial agents in response to this ongoing threat. However, pathogens continue to acquire numerous resistance mechanisms and proliferate. Meanwhile, the COVID-19 pandemic has prompted us to recognize that focusing solely on the killing strategies of immunology and microbiology in complex and multisystemic infectious diseases is insufficient for understanding survival. In essence, there is a need for a treatment approach centered on disease tolerance, capable of alleviating pathology and/or promoting physiological functions in conjunction with the existing strategies of host immune response and pathogen destruction. Against this backdrop, this narrative review comprehends and introduces microbiome therapeutics, which recently garnered attention as next-generation treatments regarding two defense strategies against infectious diseases. First, we delineate the application of existing probiotics and then the microbiome in infectious diseases. Subsequently, we expand this concept to encompass host microbiome crosstalk and present related microbiome therapeutics' current status and direction. Ultimately, the review presents a fresh perspective on understanding microbiome therapeutics in infectious diseases based on a cooperative defense system.
... Regarding functionality evaluation, tolerance to the acidic environment of the stomach is a desirable requirement of putative probiotics to reach the intestine and ultimately elicit beneficial effects 43 . The adherence of a strain to the gut epithelium is closely associated with colonization and eventually the expression of beneficial functions 44 . Our strain showed notable survivability after passage of the gastric-and duodenum-like environments and a similar attachment ratio to intestinal epithelial cells, comparable to that of the representative commercial probiotic strain, LGG, in an in vitro study (Tables S5, S6). ...
Mucosal tissues serve as the first defense line and their commensal microbiota play a role in sustaining of host health. This study aimed to isolate and evaluate a putative probiotic strain on various mucosal regions. Lactobacillus sakei HEM 224 was isolated from traditional Korean kimchi and identified. In the safety assessment L. sakei HEM 224 showed negative results for hemolysis, biogenic amine production and transferable antibiotic resistance. The probiotic potential of strain HEM 224 in diverse mucosal areas was shown in two different models, viz. a murine model with colitis induced by dextran sulfate sodium (DSS) and an allergic airway inflammation model induced by ovalbumin (OVA). In the colitis model, oral administration of L. sakei HEM 224 improved colitis physiology with immunomodulation, enhancing barrier components and gut microbiota alteration. In the allergic airway inflammation model, the intranasal administration of the strain decreased type 2 inflammation and enhanced epithelial barrier integrity from the airways. These results demonstrate that L. sakei HEM 224 can ameliorate inflammatory conditions in both the gastrointestinal and respiratory tracts through the reinforcement of the epithelial barrier and immunomodulation.
... it has been proposed that adhesion depends on the chemical and physicochemical composition of 333 the cell surface of the probiotic strain, as a result of the equilibrium of electrostatic charge and Van 334 der Waals interactions on the host's surface (Duary et al., 2011;Melo-Pereira et al., 2018). 335 ...
... Adhesion to epithelial cells is complex because two membranes cells are involved: host and microbial. Then, it has been proposed that adhesion depends on the chemical and physicochemical composition of the cell surface of the probiotic strain, as a result of the equilibrium of electrostatic charge and Van der Waals interactions on the host's surface (Duary et al., 2011;Melo-Pereira et al., 2018). Furthermore, the mechanisms of bacterial adhesion are multiple. ...
... Typical probiotics are represented by lactic acid bacteria (LAB), such as certain strains belonging to the genus Lactobacilli, Streptococci and Bifidobacterium (Haro et al., 2018;Lemme-Dumit et al., 2018). The beneficial properties of probiotic LAB on humans have some limitations related to the LAB lability and susceptibility to environmental stressors (e.g., temperature changes, dehydration, and the hostile gut environment) (Duary et al., 2011;Pedraza-Reyes et al., 2012;Botta et al., 2014;Gomathi et al., 2014). Other types of human probiotic bacteria that successfully overcome these concerns are represented by spore-forming bacteria of the Bacillus genus (e.g., Bacillus subtilis, Bacillus coagulans, and Bacillus clausii) (Sanders et al., 2003;Kalman et al., 2009;Cutting, 2011;Hanifi et al., 2015;Piewngam et al., 2018;Piewngam and Otto, 2020;Freedman et al., 2021;Rodriguez Ayala et al., 2021;Zeng et al., 2021). ...
... Probiotic bacteria do not always produce in vivo beneficial effects for the host. The ability to reach and colonize (in the desired number) the GI tract is sometimes compromised (Duary et al., 2011;Botta et al., 2014;Gomathi et al., 2014). For example, LAB probiotics have shown differences in their ability to survive GI conditions depending on the source of their isolation, preservation form, or packaging (e.g., powdered, or liquid form, microencapsulated, lyophilized) (Ozawa et al., 2012). ...
Spore-forming probiotic bacteria offer interesting properties as they have an intrinsic high stability, and when consumed, they are able to survive the adverse conditions encountered during the transit thorough the host gastrointestinal (GI) tract. A traditional healthy food, nattō, exists in Japan consisting of soy fermented by the spore-forming bacterium Bacillus subtilis natto. The consumption of nattō is linked to many beneficial health effects, including the prevention of high blood pressure, osteoporosis, and cardiovascular-associated disease. We hypothesize that the bacterium B. subtilis natto plays a key role in the beneficial effects of nattō for humans. Here, we present the isolation of B. subtilis DG101 from nattō and its characterization as a novel spore-forming probiotic strain for human consumption. B. subtilis DG101 was non-hemolytic and showed high tolerance to lysozyme, low pH, bile salts, and a strong adherence ability to extracellular matrix proteins (i.e., fibronectin and collagen), demonstrating its potential application for competitive exclusion of pathogens. B. subtilis DG101 forms robust liquid and solid biofilms and expresses several extracellular enzymes with activity against food diet-associated macromolecules (i.e., proteins, lipids, and polysaccharides) that would be important to improve food diet digestion by the host. B. subtilis DG101 was able to grow in the presence of toxic metals (i.e., chromium, cadmium, and arsenic) and decreased their bioavailability, a feature that points to this probiotic as an interesting agent for bioremediation in cases of food and water poisoning with metals. In addition, B. subtilis DG101 was sensitive to antibiotics commonly used to treat infections in medical settings, and at the same time, it showed a potent antimicrobial effect against pathogenic bacteria and fungi. In mammalians (i.e., rats), B. subtilis DG101 colonized the GI tract, and improved the lipid and protein serum homeostasis of animals fed on the base of a normal- or a deficient-diet regime (dietary restriction). In the animal model for longevity studies, Caenorhabditis elegans , B. subtilis DG101 significantly increased the animal lifespan and prevented its age-related behavioral decay. Overall, these results demonstrate that B. subtilis DG101 is the key component of nattō with interesting probiotic properties to improve and protect human health.
... To adjudge the colonization capability of a potential probiotic strain, adhesion has been considered an ideal parameter. The adhesion capability of a Lactobacillus strain is the determining probiotic attribute in several studies (Bhat et al., 2019;Munoz et al., 2011;Duary et al., 2011;Vesterlund et al., 2005). However, reports showed that the colonization by the probiotics was not permanent rather once the consumption was ceased it resulted in the disappearance of the microbe in the feces (Bennet et al., 1992). ...
... Thus, yeasts with an SHb above 35% were selected. The hydrophobicity of the cell surface allows a better interaction between the microorganism and the epithelial cells of the GIT (Duary et al. 2011). Yeasts may have an advantage in eliminating pathogenic bacteria through competition for space since the cells of yeasts are usually larger than bacteria. ...
The use of yeasts as a feed supplement for cattle can promote animal development and performance. However, for the positive results to be consistent, strains with probiotic properties must be selected. The objective of this study was to isolate and identify yeasts present in the bovine feces and evaluate their probiotic potential together with strains previously isolated from the rumen (preliminary study). A total of 193 isolates were studied, including 139 isolates (19 species) from fecal samples from 11 different animals (Bos taurus and Bos indicus) and 54 strains previously isolated from rumen fluid (Bos taurus). The yeast population in the feces ranged from 3.51 to 4.99 log CFU/g, with Candida pararugosa being the most abundant (isolated from the feces of six samples analysed). Isolates were selected that had negative results in the safety tests (hemolytic activity, DNAse, and gelatinase) and had percentages greater than 35 and 70% for hydrophobicity and auto-aggregation, respectively. In addition, selected isolates had percentages greater than 77.7 and 74.7% for coaggregation with pathogenic strains of Escherichia coli and Clostridium perfringens, respectively. The isolates with percentage growth at 39 °C greater than 64.6% and viability greater than 96.7% were selected for survival testing under bovine gastrointestinal conditions. After the tests, the seven best isolates were selected, belonging to the species Candida pararugosa (L60, CCMA 928 and CCMA 930) and Pichia kudriavzevii (L97, L100, CCMA904, CCMA 907). The selected isolates were exopolysaccharide producers. Based on the results of the evaluated properties, the seven selected isolates were classified as potential probiotics for cattle.
