Probiotics and Antimicrobial Proteins最新文献

Live micro-ecological agents, such as probiotics, have demonstrated a significant role in the preservation of human health, encompassing oral health maintenance and regulation of oral microbiota. Here, a total of 20 patients diagnosed with chronic periodontitis were recruited and randomly assigned into two cohorts based on completion of physiotherapy: a placebo group (n = 10) and a probiotic group (n = 10). The actual efficacy was assessed by administering chewable tablets (5 × 10⁹ CFU/tablet) containing the probiotics Lactobacillus salivarius LS97, Lactobacillus paracasei LC86, and Lactobacillus acidophilus LA85 to patients with chronic periodontitis. For the placebo group, chewable tablets without probiotics were administered, while maintaining consistency with the rest of the ingredients used in the probiotic group. Saliva and plaque samples were collected at different time points (0, 1, and 3 months) and subjected to 16S amplicon sequencing for microbial structure analysis. Salivary IgA content was determined using enzyme immunoassay, whereas clinical chronic periodontal pocket depth (PD) and bleeding on probe index (BOP +) were employed to evaluate the actual efficacy of probiotic-assisted physiological intervention in chronic periodontitis treatment. Compared to the placebo group, the probiotic intervention resulted in a significant increase in salivary IgA levels among patients, accompanied by a notable decrease in PD and BOP + levels. Furthermore, the probiotic intervention led to a substantial reduction in Fusobacterium and Porphyromonas counts, while significantly increasing Lactobacillus abundance within the dental plaque microbiota of patients. Importantly, no significant alterations were observed in the overall structure of both salivary and dental plaque microbiota following the probiotic intervention. The administration of this live probiotic agent consistently and significantly enhances the oral immune response in patients with chronic periodontitis, thereby augmenting the effectiveness of physical interventions for this condition. Moreover, it effectively reduces the abundance of pathogenic microbes associated with chronic periodontitis without causing substantial alterations to the salivary and dental plaque microbiota composition. Trial registration: Chinese Clinical Trial Registry (ChiCTR) ( https://www.chictr.org.cn ) under the registration number ChiCTR2300074108.

This study isolated and characterized four indigenous lactic acid bacterial strains from naturally fermented green tender coconut water: Lactiplantibacillus plantarum CWJ3, Lacticaseibacillus casei CWM15, Lacticaseibacillus paracasei CWKu14, and Lacticaseibacillus rhamnosus CWKu-12. Notably, among the isolates, Lact. plantarum CWJ3 showed exceptional acid tolerance, with the highest survival rate of 37.34% at pH 2.0 after 1 h, indicating its higher resistance against acidic gastric conditions. However, all strains exhibited robust resistance to bile salts, phenols, and NaCl, with survival rates exceeding 80% at given concentrations. Their optimal growth at 37 °C and survival at 20 °C and 45 °C underscored adaptability to diverse environmental conditions. Additionally, all strains showed sustainable survival rates in artificial saliva and simulated gastrointestinal juices, with Lact. plantarum CWJ3 exhibiting significantly higher survival rate (70.66%) in simulated gastric juice compared to other strains. Adherence properties were particularly noteworthy, especially in Lact. rhamnosus CWKu-12, which demonstrated the highest hydrophobicity, coaggregation with pathogens and autoaggregation, among the strains. The production of exopolysaccharides, particularly by Lact. plantarum CWJ3, enhanced their potential for gut colonization and biofilm formation. Various in vitro antioxidative assays using spectrophotometric methods revealed the significant activity of Lact. plantarum CWJ3, while antimicrobial testing highlighted its efficacy against selected foodborne pathogens. Safety assessments confirmed the absence of biogenic amine production, hemolytic, DNase, and gelatinase activities, as well as the ability to hydrolase the bile salt. Furthermore, these non-dairy probiotics exhibited characteristics comparable to dairy derived probiotics, demonstrating their potential suitability in developing novel probiotic-rich foods and functional products.

Human milk is the best nutrition for infants, providing optimal support for the developing immune system and gut microbiota. Hence, it has been used as source for probiotic strain isolation, including members of the genus Bifidobacterium, in an effort to provide beneficial effects to infants who cannot be exclusively breastfed. However, not all supplemented bifidobacteria can effectively colonise the infant gut, nor confer health benefits to the individual infant host; therefore, new isolates are needed to develop a range of dietary products for this specific age group. Here, we investigated the beneficial potential of Bifidobacterium breve DSM 32583 isolated from human milk. We show that in vitro B. breve DSM 32583 exhibited several characteristics considered fundamental for beneficial bacteria, including survival in conditions simulating those present in the digestive tract, adherence to human epithelial cell lines, and inhibition of growth of potentially pathogenic microorganisms. Its antibiotic resistance patterns were comparable to those of known beneficial bifidobacterial strains, and its genome did not contain plasmids nor virulence-associated genes. These results suggest that B. breve DSM 32583 is a potential probiotic candidate.

Graphical Abstract

Probiotic bacteria, and especially lactic acid bacteria, have long been known to wield a variety of health-beneficial effects, including antioxidant, antimicrobial, anti-inflammatory, immunomodulatory, and anticancer activities. However, our understanding of the mechanisms involved in these activities remains incomplete. In this study, we wished to investigate the processes that give rise to the anticancer activity of Lacticaseibacillus casei ATCC393 and the possibility that immunogenic cell death of cancer cells can be induced following treatment with this probiotic. In both cell lines that we have examined, we detected notable pro-apoptotic signaling, including the upregulation of death receptors, that culminated in the activation of caspase 3, the endpoint and most characteristic effector molecule of all pro-apoptotic cascades. In addition, we identified damage-associated molecular patterns associated with immunogenic cell death. Calreticulin exposure on the outer cell membrane, HMGB1 translocation outside the nucleus and depletion of intracellular ATP was evident in both cancer cell lines treated with the probiotic, while expression of type I interferons was upregulated in CT26 cells. Our findings suggest that treatment with the probiotic induced apoptosis in cancer cells, mediated by extrinsic death receptor signaling. Moreover, it resulted in the release of molecular signals related with immunogenic cell death and induction of cancer cell-specific adaptive immune responses.

Multidrug-resistant Escherichia coli (MDR-E. coli) is a global health concern. Lactic acid bacteria (LAB) are important probiotics that have beneficial effects on health, and in recent years, their influences in preventing foodborne pathogens-induced colitis have attracted much attention. Therefore, this study aimed to investigate the oral administration of Lactiplantibacillus plantarum NWAFU-BIO-BS29 as an emerging approach to alleviate MDR-E. coli–induced colitis in BALB/c mice model. To illustrate the mode of action of NWAFU-BIO-BS29 interventions with the gut microbiota and immune responses, the changes on the colonic mucosal barrier, regulatory of the gene expressions of inflammatory cytokines, re-modulating the intestinal microflora, and changes in physiological parameters were studied. The results indicated that daily supplementation of 200 µL fresh bacteria for 7 days had ameliorated the associated colitis and partially prevented the infection. The modes of action by ameliorating the inflammatory response, which destructed villous and then affected the intestinal barrier integrity, reducing the secretion of interleukins (6 and β) and tumor necrosis factor (TNF-α) in serum by 87.88–89.93%, 30.73–35.98%, and 19.14–22.32%, respectively, enhancing the expressions of some epithelial integrity-related proteins in the mouse mucous layer of mucins 2 and 3, Claudin-1, and Occludin by 130.00–661.85%, 27.64–57.35%, 75.52–162.51%, and 139.36–177.73%, respectively, and 56.09–73.58% for toll-like receptor (TLR4) in colon tissues. Notably, the mouse gut microbiota analysis showed an increase in the relative abundance of beneficial bacteria, including Lactobacillus, Bacteriodales bacterium, Candidatus Saccharimonas, Enterorhabdus, and Bacilli. Furthermore, the probiotic promoted the proliferation of epithelia and goblet cells by increasing short-chain fatty acids (SCFAs) levels by 19.23–31.39%. In conclusion, L. plantarum NWAFU-BIO-BS29 has potential applications and can be considered a safe dietary supplement to ameliorate the colitis inflammation symptoms of MDR-E. coli infection.

Biofilms may contain pathogenic and spoilage bacteria and can become a recurring problem in the dairy sector, with a negative impact on product quality and consumer health. Peracetic acid (PAA) is one of the disinfectants most frequently used to control biofilm formation and persistence. Though effective, it cannot be used at high concentrations due to its corrosive effect on certain materials and because of toxicity concerns. The aim of this study was to test the possibility of PAA remaining bactericidal at lower concentrations by using it in conjunction with reuterin (3-hydroxypropionaldehyde). We evaluated the efficacy of PAA in pure form or as BioDestroy^®, a PAA-based commercial disinfectant, on three-species biofilms formed by dairy-derived bacteria, namely Pseudomonas azotoformans PFlA1, Serratia liquefaciens Sl-LJJ01, and Bacillus licheniformis Bl-LJJ01. Minimum inhibitory concentrations of the three agents were determined for each bacterial species and the fractional inhibitory concentrations were then calculated using the checkerboard assay. The minimal biofilm eradication concentration (MBEC) of each antibacterial combination was then calculated against mixed-species biofilm. PAA, BioDestroy^®, and reuterin showed antibiofilm activity against all bacteria within the mixed biofilm at respectively 760 ppm, 450 ppm, and 95.6 mM. The MBEC was lowered significantly to 456 ppm, 337.5 ppm, and 71.7 mM, when exposed to reuterin for 16 h followed by contact with disinfectant. Combining reuterin with chemical disinfection shows promise in controlling biofilm on food contact surfaces, especially for harsh or extended treatments. Furthermore, systems with reuterin encapsulation and nanotechnologies could be developed for sustainable antimicrobial efficacy without manufacturing disruptions.

This study investigates the postbiotic potential of extracellular products (ECPs) from Bacillus pumilus strains cultivated on microalgae-supplemented media. We assessed enzymatic and antimicrobial activities to select ECPs that enhance the digestive processes in gilthead seabream. Additionally, we explored the in vitro enzymatic capacity of the chosen postbiotics to hydrolyze macromolecules in microalgae. Finally, a feeding trial was conducted to determine the in vivo effects of the ECPs on Sparus aurata. In vitro enzymatic assays demonstrated diverse hydrolytic capacities among ECPs. All conditions exhibited antimicrobial activity against Photobacterium damselae subsp. piscicida, with variation in inhibitory effects against Vibrio harveyi and Tenacibaculum maritimum. Furthermore, in vitro assays revealed differences in protein hydrolysis and soluble protein concentration, influencing amino acid and reducing sugar release from microalgal biomass. These analyses facilitated a selection to test ECPs in vivo. Lastly, the in vivo experiment revealed no differences in the growth performance, nutrient utilization, and general metabolism of S. aurata fed the experimental diets. Dietary inclusion of postbiotics increased the activity of key digestive enzymes in fish compared to the control group, and particularly, values increased significantly when the fish were fed with the ECP-nanoparticulate-supplemented diet. In conclusion, the inclusion of microalgae in the culture media significantly influences the activity of extracellular products from B. pumilus strains, as evidenced in both in vitro and in vivo assays.

Intermittent fasting (IF) is an increasingly popular dietary approach involving alternating fasting and eating periods. This review aims to summarize the growing body of literature demonstrating that IF is a potential nutritional practice that involves alternating periods of fasting and eating and the numerous benefits of IF, especially in the modulation of healthy gut microbiota. The positive impact of intermittent fasting on gut microbiota not only promotes gastrointestinal health but also has far-reaching effects on critical systems throughout the body. Additionally, the evidence presented in this review highlights the significant preventive and therapeutic effects of intermittent fasting on a wide range of disorders. This includes reducing the risk of diabetes, and neurological disorders, alleviating obesity, and enhancing the functioning of the liver, ultimately contributing to the maintenance of metabolic equilibrium. Perhaps most notably, these effects play a substantial role in preventing diabetes, a global health concern of increasing significance. This comprehensive investigation delves into the scientific foundations of intermittent fasting’s impact on gut microbiota and its implications for averting chronic diseases, providing valuable insights for future research and therapeutic applications.

Helicobacter pylori infects the gastric mucosa and induces chronic gastritis, peptic ulcers, and gastric cancer. Research has demonstrated that vaccination can induce a protective immune response and prevent H. pylori infection. Oral administration of the Lactococcus lactis live-carrier vaccine is safe and easily complied with by the public. In this study, two recombinant L. lactis strains were constructed that expressed antigens of H. pylori urease subunit alpha (UreA) and UreA fused with Escherichia coli heat-labile toxin B subunit (LTB-UreA), named LL-UreA and LL-LTB-UreA, respectively. The expression of antigen proteins was confirmed by Western blotting analysis. Survival assessment indicated that the engineered L. lactis could colonize in the digestive tract of BALB/c mice up to 10 days after the last oral administration with our immunization protocol. The ability to induce immune response and immune protective efficacy of the L. lactis were confirmed. These results indicated that oral administration with LL-UreA or LL-LTB-UreA could induce UreA-specific mucosal secretory IgA (sIgA) and cellular immune response, significantly increasing the cytokines levels of interferon-gamma (IFN-γ), interleukin (IL)-17A, and IL-10, together with the proportion of CD4⁺IFN-γ⁺ T cells and CD4⁺IL17A⁺ T cells. More importantly, oral administration of LL-UreA and LL-LTB-UreA brought about effective protection in mice to prevent H. pylori infection, especially LL-UreA, resulting in 70% of mice showing no H. pylori colonization and the remaining 30% showing only low levels of colonization. These findings underscore the potential of using orally administered engineered L. lactis vaccines to prevent H. pylori infection.

Plant defensins are small antimicrobial proteins (AMP) that participate in the immune defense of plants through their antibacterial, antiviral and antifungal activities. PgD1 is a defensin from Picea glauca (Canadian Pine) and has antifungal activity against plant pathogens. This activity positions it as an alternative biotechnological agent to pesticides commonly used against these plant fungi diseases. The present study aimed to recombinantly produce PgD1 in Escherichia coli to characterize its in vitro antifungal potential against different phytopathogens. To achieve this, the coding gene was amplified and cloned into pET30a( +). Recombinant plasmid was subsequently introduced into E. coli for the soluble expression of defensin PgD1. To evaluate the antifungal activity of the expressed protein, the growth inhibition test was used in solid and liquid media for approximately 7 days against significant plant pathogens, that cause significant crop damage including: Botrytis cinerea, Colletotrichum gloeosporioides, Colletotrichum musae, Colletotrichum graminicola and Fusarium oxysporum. Additionally, stability assessments included temperature variation experiments and inhibition tests using dithiothreitol (DTT). The results showed that there was significant inhibition of the fungal species tested when in the presence of PgD1. Furthermore, defensin proved to be resistant to temperature variations and demonstrated that part of its stability is due to its primary structure rich in cysteine ​​residues through the denaturation test with dithiothreitol (DTT) where the antifungal activity of PgD1 defensin was inhibited. These data indicate that recombinant PgD1 could be utilized as a plant protection technology in agriculture.