Vibriocidal efficacy of Bifidobacterium bifidum and Lactobacillus acidophilus cell-free supernatants encapsulated in chitosan nanoparticles against multi-drug resistant Vibrio cholerae O1 El Tor.

BACKGROUND: Cholera is a diarrheal disease recognized for being caused by toxin-producing Vibrio (V.) cholerae. This study aims to assess the vibriocidal and immunomodulatory properties of derived cell-free supernatants (CFSs) of Bifidobacterium (B.) bifidum and Lactobacillus (L.) acidophilus encapsulated in chitosan nanoparticles (CFSb-CsNPs and CFSa-CsNPs) against clinical multi-drug resistance (MDR) isolates of V. cholerae O1 El Tor. METHODS: We synthesized CFSb-CsNPs and CFSa-CsNPs using the ionic gelation technique. The newly nanostructures were characterized for size, surface zeta potential, morphology, encapsulation efficacy (EE), stability in different pH values and temperatures, release profile, and in vitro cytotoxicity. The antimicrobial and antibiofilm effects of the obtained nanocomposites on clinical MDR isolates (N = 5) of V. cholerae E1 Tor O1 were investigated by microbroth dilution assay and crystal violet staining, respectively. We conducted quantitative real-time PCR (qRT-PCR) to analyze the relative gene expressions of Bap, Rbmc, CTXAB, and TCP in response to CFSb-CsNPs and CFSa-CsNPs. Additionally, the immunomodulatory effects of formulated structures on the expression of TLR2 and TLR4 genes in human colorectal adenocarcinoma cells (Caco-2) were studied. RESULTS: Nano-characterization analyses indicated that CFSb-CsNPs and CFSa-CsNPs exhibit spherical shapes under scanning electron microscopy (SEM) imaging, with mean diameters of 98.16 ± 0.763 nm and 83.90 ± 0.854 nm, respectively. Both types of nanoparticles possess positive surface charges. The EE% of CFSb-CsNPs was 77 ± 4.28%, whereas that of CFSa-CsNPs was 62.5 ± 7.33%. Chitosan (Cs) encapsulation leads to increased stability of CFSs in simulated pH conditions of the gastrointestinal tract in which the release rates for CFSb-CsNPs and CFSa-CsNPs were reached at 58.00 ± 1.24% and 55.01 ± 1.73%, respectively at pH = 7.4. The synergistic vibriocidal effects observed from the co-administration of both CFSb-CsNPs and CFSa-CsNPs, as evidenced by a fractional inhibitory concentration (FIC) index of 0.57, resulting in a significantly lower MIC of 2.5 ± 0.05 mg/mL (p < 0.0001) compare to individual administration. The combined antibacterial effect of CFSb-CsNPs and CFSa-CsNPs on Bap (0.14 ± 0.05), Rbmc (0.24 ± 0.01), CTXAB (0.30 ± 0.09), and TCP (0.38 ± 0.01) gene expression was significant (p < 0.001). Furthermore, co-administration of CFSb-CsNPs and CFSa-CsNPs also demonstrated the potency of suppressing TLR 2/4 (0.20 ± 0.01 and 0.12 ± 0.02, respectively) gene expression (p = 0.0019) and reduced Caco-2 cells attached bacteria to 526,000 ± 51,46 colony-forming units/mL (11.19%) (p < 0.0001). CONCLUSION: Our study revealed that encapsulating CFSs within CsNPs enhances their vibriocidal activity by improving stability and enabling a controlled release mechanism at the site of interaction between the host and bacteria. Additionally, the simultaneous use of CFSb-CsNPs and CFSa-CsNPs exhibited superior vibriocidal potency against MDR V. cholerae O1 El Tor strains, indicating these combinations as a potential new approach against MDR bacteria.

Mapping a sustainable approach: biosynthesis of lactobacilli-silver nanocomposites using whey-based medium for antimicrobial and bioactivity applications.

This study explores a sustainable approach for synthesizing silver nanocomposites (AgNCs) with enhanced antimicrobial and bioactivity using safe Lactobacillus strains and a whey-based medium (WBM). WBM effectively supported the growth of Lactobacillus delbrueckii and Lactobacillus acidophilus, triggering a stress response that led to AgNCs formation. The synthesized AgNCs were characterized using advanced spectroscopic and imaging techniques such as UV‒visible, Fourier transform infrared (FT-IR) spectroscopy, transmission electron (TEM), and scanning electron microscopy with energy dispersive X-ray analysis (SEM-Edx). Lb acidophilus-synthesized AgNCs in WBM (had DLS size average 817.2-974.3 ± PDI = 0.441 nm with an average of metal core size 13.32 ± 3.55 nm) exhibited significant antimicrobial activity against a broad spectrum of pathogens, including bacteria such as Escherichia coli (16.47 ± 2.19 nm), Bacillus cereus (15.31 ± 0.43 nm), Clostridium perfringens (25.95 ± 0.03 mm), Enterococcus faecalis (32.34 ± 0.07 mm), Listeria monocytogenes (23.33 ± 0.05 mm), methicillin-resistant Staphylococcus aureus (MRSA) (13.20 ± 1.76 mm), and filamentous fungi such as Aspergillus brasiliensis (33.46 ± 0.01 mm). In addition, Lb acidophilus-synthesized AgNCs in WBM exhibit remarkable free radical scavenging abilities, suggesting their potential as bioavailable antioxidants. These findings highlight the dual functionality of these biogenic AgNCs, making them promising candidates for applications in both medicine and nutrition.

The effect of arginine on the growth of probiotics.

OBJECTIVE(S): The study objective was to examine the effect of arginine (Arg) supplementation on the growth of probiotics. METHODS: Lacticaseibacillus rhamnosus GG, Lactiplantibacillus plantarum, and Lactobacillus acidophilus were identified as potential probiotics. L. rhamnosus GG and L. plantarum were selected for further experimentation. The probiotics were co-treated with 0.9 % NaCl (negative control), 0.5 % Arg, and 1.0 % Arg in a 1:1 ratio for 24 h at 5 % CO2, 37 °C. The probiotics were tested for growth profiles, spectroscopic turbidity assay, metabolic assays (XTT and WST-8), live/dead cell assessment using confocal laser scanning microscopy (CLSM), and colony forming units (CFU). RESULTS: The growth profiles of L. rhamnosus GG and L. plantarum were found to be similar, whereas L. acidophilus showed minimal or no transition from the initial lag phase. In the turbidity assay, the end-point absorbance for L. rhamnosus GG with 1.0 % Arg was significantly lower than 0.9 % NaCl and 0.5 % Arg (p < 0.05). For metabolic assays and CFU, increasing concentrations of Arg increased the viable cells for L. rhamnosus GG (p < 0.05), but decreased viability for L. plantarum (p < 0.05). Metabolic assays with dual-species bacterial suspensions indicated that Arg co-treatment inhibited viable proportions compared to control (p < 0.05). The dead cell proportion was significantly lower than live cell proportion for all tested interventions and probiotics (p < 0.05). CONCLUSION: Increasing concentrations of Arg promote the growth of L. rhamnosus GG, while conversely inhibiting the growth of L. plantarum. Therefore, the effect of prebiotic Arg on probiotics is concentration-dependent, leading to a selective promotion or inhibition of growth. CLINICAL SIGNIFICANCE: The present study results show that Arg supplementation can selectively enhance the growth of L. rhamnosus GG while inhibit the growth of L. plantarum. This underscores the need to consider strain-specific responses in probiotic formulations when developing Arg-based synbiotics for modulating biofilms and creating ecologically homeostatic biofilm microenvironments.

Evaluating antimicrobial activity and cytotoxicity of silver nanoparticles incorporated into reinforced zinc oxide eugenol: an in vitro study.

PURPOSE: This study aimed to evaluate the antibacterial and cytotoxic effects of reinforced zinc oxide-eugenol (rZOE) incorporated with different concentrations of silver nanoparticles (AgNPs). METHODS: The pastes of rZOE alone or mixed with AgNPs at concentrations of 1%, 2%, and 5% of weight were prepared. In vitro antimicrobial activity of prepared materials against Streptococcus (S.) mutans and Lactobacillus (L.) acidophilus were evaluated after 2, 4, and 6 h of contact times using direct contact test (DCT) and also following 24 h incubation by well-diffusion test (WDT). The cytotoxicity of the tested materials on human dental pulp stem cells was also determined by MTT assay. RESULTS: The DCT demonstrated that the time-dependent reductions of the colony numbers of both bacteria by three different concentrations of AgNPs incorporated into rZOE were equal but steeper than the rZOE alone (P < 0.05). The increases in growth inhibition zones of S. mutans and L. acidophilus were associated with the increasing concentration of AgNPs mixed with rZOE in the WDT; however, statistical analysis did not show any significant differences (P = 0.092). The MTT assay revealed a significantly lower percentage of cell viability after 1 day of culture only with the rZOE + AgNP5% in comparison to the rZOE alone (P = 0.011) and the control medium (P = 0.001). CONCLUSION: Since the antimicrobial activities of three different concentrations of AgNPs incorporated into rZOE were equal and AgNPs had lower toxicity at lower concentrations, using AgNPs at 1% concentration is suggested to be mixed with rZOE.

Bio-inspired synthesis of silver nanoparticles usingSalsola imbricataand its application as antibacterial additive in glass ionomer cement.

Nanotechnology has gained immense popularity and observed rapid development due to the remarkable physio-chemical properties of nanoparticles (NPs) and related nanomaterials. The green production of NPs has many benefits over traditional techniques because the current procedures are expensive, time-consuming, and involve harmful substances that limit their applicability. This study aimed to use a novel green source, theSalsola imbricata(SI) plant, which is commonly found in Central Asia and known for its medicinal properties as a reducing and stabilizing agent for the synthesis of AgNPs. The current study also utilized efficient statistical design, the Plackett-Burman Design (PBD) of Experiment method to synthesize the NPs. The characterization of NPs was carried out using UV-Vis spectroscopy, Fourier transform infrared spectroscopy, and scanning electron microscopy (SEM). The PBD results showed that only two out of four factorsi.e.AgNO3concentration and incubation time, were significant for the synthesis of SI-AgNPs. While remaining factors, incubation temperature and plant extract: AgNO3ratio were non-significant. The SEM analysis result showed that SI-AgNPs had a size of 20-50 nm. The SI-AgNPs demonstrated strong antibacterial activity against oral pathogens such asS. mutans and Lactobacillus acidophilus, with the highest efficacy observed at a concentration of 2 mg ml-1. The addition of SI-AgNPs in glass ionomer cement significantly increased the antibacterial activity of GIC againstS. mutans. Based on the results of the current study, the plant based AgNPs can be further evaluated in detail as alternate antimicrobial agent either alone or in combination with other antimicrobial agents for different dental applications.

Enhanced probiotic viability in innovative double-network emulsion gels: Synergistic effects of the whey protein concentrate-xanthan gum complex and κ-carrageenan.

In this study, the whey protein concentrate and xanthan gum complex obtained by specific pH treatment, along with κ-carrageenan (KC), were used to encapsulate Lactobacillus acidophilus JYLA-191 in an emulsion gel system. The effects of crosslinking and KC concentration on the visual characteristics, stability, mechanical properties, and formation mechanism of emulsion gels were investigated. The results of optical imaging, particle size distribution, and rheology exhibited that with the addition of crosslinking agents, denser and more homogeneous emulsion gels were formed, along with a relative decrease in the droplet size and a gradual increase in viscosity. Especially when the concentration of citric acid (CA) was 0.09 wt%, KC was 0.8 wt%, and K+ was present in the system, the double-network emulsion gel was stable at high temperatures and in freezing environments, and the swelling ratio was the lowest (9.41%). Gastrointestinal tract digestive treatments and pasteurization revealed that the probiotics encapsulated in the double-network emulsion gel had a higher survival rate, which was attributed to the synergistic cross-linking of CA and K+ biopolymers to construct the emulsion gels. Overall, this study highlights the potential of emulsion gels to maintain probiotic vitality and provides valuable insights for developing inventive functional foods.

Inhibitory effects of propolis and essential oils on oral bacteria.

INTRODUCTION: Propolis is a natural composite balsam. In the past decade, propolis has been extensively investigated as an adjuvant for the treatment of periodontitis. This study aimed to investigate antimicrobial activities of propolis solutions and plant essential oils against some oral cariogenic (Streptococcus mutans, Streptococcus mitis, Streptococcus sanguis, Lactobacillus acidophilus) and periodontopathic bacteria (Actinomyces odontolyticus, Eikenella corrodens, Fusobacterium nucleatum). METHODOLOGY: Determination of the minimum inhibitory concentration (MIC): The antimicrobial activity of propolis and essential oils was investigated by the agar dilution method. Serial dilutions of essential oils were prepared in plates, and the assay plates were estimated to contain 100, 50, 25 and 12.5 µg/mL of active essential oils. Dilutions for propolis were 50, 25, 12.5 and 6.3 µg/mL of active propolis solutions. RESULTS: Propolis solutions dissolved in benzene, diethyl ether and methyl chloride, demonstrated equal effectiveness against all investigated oral bacteria (MIC=12.5 µg/mL). Propolis solution dissolved in acetone displayed MIC of 6.3 µg/mL only for Lactobacillus acidophilus. At the MIC of 12.5 µg/mL, essential oils of Salvia officinalis and Satureja kitaibelii were effective against Streptococcus mutans and Porphyromonas gingivalis, respectively. For the latter, the MIC value of Salvia officinalis was twice higher. CONCLUSIONS: The results indicate that propolis and plant essential oils appear to be a promising source of antimicrobial agents that may prevent dental caries and other oral infectious diseases.

Antibiofilm effects of Thymus vulgaris and Hyptis spicigera essential oils on cariogenic bacteria.

Aim: The inhibitory and antibiofilm effects of Thymus vulgaris (EOTv) and Hyptis spicigera essential oils (EOHs) on cariogenic microorganisms were evaluated. Materials & methods: The chemical characterization of EOTv was performed by gas chromatography/mass spectrometry. Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Streptococcus mitis, Streptococcus sobrinus, Lactobacillus acidophilus and Actinomyces naeslundii were used for agar diffusion assays and determination of minimal inhibitory and minimal bactericide concentrations. In addition, 20 streptococci and lactobacilli clinical isolates were also tested. The effects of essential oil on microbial initial biofilm formation and on preformed microcosm biofilm formed from human saliva were studied. Results & conclusion: Both essential oils had inhibitory effects on the cariogenic species and reduced the bacterial adherence to dental enamel. Essential oils were able to disrupt preformed microcosm biofilms. Thymus vulgaris and Hyptis spicigera essential oils have potential to be used in the development of formulations to the control of cariogenic biofilms.

Promotive effects of sesamin on proliferation and adhesion of intestinal probiotics and its mechanism of action.

The effect of sesamin on intestinal flora was studied by in vitro animal fecal anaerobic fermentation system, and were analyzed by 16S rDNA sequencing. Results showed that sesamin modulated the composition of intestinal microorganisms and reshaped gut microbiome. The abundance of probiotics Lactobacillaceae and Bifidobacteriaceae increased, while the abundance of Enterobacteriaceae decreased. The properties of probiotics (Bifidobacterium bifidum and Lactophilus acidophilus) adhesion to epithelial colon cells (NCM460) were assessed by gram staining and plate counting methods. Results showed that sesamin increased the adhesive index of probiotics. Analysis of RT-qPCR, Western blot and immunofluorescence staining indicated that sesamin up-regulated the expression of the adhesive protein (ß-cadherin and E-cadherin) of NCM460 cells. In conclusion, sesamin could promote the proliferation and adhesion of intestinal probiotics leading to modulating gut microbiota, which provided basis for sesamin as a food-borne functional factor for improving intestinal health.

Protective effect of the probiotic Lactobacillus acidophilus ATCC 4356 in BALB/c mice infected with Toxocara canis.

Human toxocariasis consists of chronic tissue parasitosis that is difficult to treat and control. This study aimed to evaluate the action of the probiotic Lactobacillus acidophilus ATCC 4356 on larvae of Toxocara canis and the effect of IFN-γ cytokine on parasite-host in vivo (1.109 CFU) and in vitro (1.106, 1.107, 1.108, 1.109 CFU) interactions. Four groups of six BALB/c mice were formed: G1 - L. acidophilus supplementation and T. canis infection; G2 - T. canis infection; G3 - L. acidophilus supplementation; and G4 - PBS administration. Mice were intragastrically suplemented with probiotics for 15 days before inoculation and 48 h after inoculation with 100 T. canis eggs. The inoculation of T. canis was also perfomed intragastrically. The recovery of larvae took place through digestion of liver and lung tissues; the evaluation of IFN-γ gene transcription in leukocytes was performed by qPCR. The in vitro test consisted of incubating the probiotic with T. canis larvae. The supplementation of probiotics produced a reduction of 57.7% (p = 0.025) in the intensity of infection of T. canis larvae in mice, whereas in the in vitro test, there was no larvicidal effect. In addition, a decrease in the IFN-γ gene transcription was observed in both, T. canis-infected and uninfected mice, regardless of whether or not they received supplementation. The probiotic L. acidophilus ATCC 4356 reduced T. canis infection intensity in mice, however, the probiotic did not have a direct effect on larvae, demonstrating the need of interaction with the host for the beneficial effect of the probiotic to occur. Yet, the proinflammatory cytokine IFN-γ did not apparently contributed to the observed beneficial effect of probiotics.

Starvation Survival and Biofilm Formation under Subminimum Inhibitory Concentration of QAMs.

Quaternary ammonium methacrylates (QAMs) are useful antimicrobial compounds against oral bacteria. Here, we investigated the effects of two QAMs, dimethylaminododecyl methacrylate (DMADDM) and dimethylaminohexadecyl methacrylate (DMAHDM), on biofilm formation, survival and development of tolerance by biofilm, and survival and development of tolerance against QAMs after prolonged starvation. Enterococcus faecalis (E. faecalis), Streptococcus gordonii (S. gordonii), Lactobacillus acidophilus (L. acidophilus), and Actinomyces naeslundii (A. naeslundii) were used. Minimum inhibitory concentration (MIC) of QAMs against multispecies biofilm was determined. Biofilm formed under sub-MIC was observed by crystal violet staining and confocal laser scanning microscopy (CLSM). Metabolic activity was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactic acid production measurement. Development of tolerance was determined by MIC values before and after exposure to QAMs or after prolonged starvation. It was found that E. faecalis and S. gordonii could survive and form biofilm under sub-MIC of QAMs. Lactic acid production from biofilms formed under sub-MIC was significantly higher than control specimens (p < 0.05). The exposure to sub-MIC of QAMs promoted biofilm formation, and prolonged starvation or prolonged contact with sub-MIC helped bacteria develop tolerance against killing by QAMs.

Chitosan capping of CuO nanoparticles: Facile chemical preparation, biological analysis, and applications in dentistry.

This investigation is vital contribution to the healthcare system utilizing techniques of nanobiotechnology. It interestingly applies chitosan capped CuO nanoparticles in the field of medicine and restorative dentistry. The CuO nanoparticles and CuO-Chitosan nanoparticles are prepared by co-precipitation, and their characterization is performed using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy dispersive X-ray (EDX). The average crystallite size of these nanoparticles has been found to be in the dimensions of <40 nm and <35 nm, respectively. CuO-Chitosan nanoparticles show significant enhancement in in vitro antibacterial, antioxidant, cytotoxic, and antidiabetic activity as compared to CuO nanoparticles. In addition, the successful amalgamation of CuO nanoparticles and CuO-Chitosan nanoparticles into dentine bonding agents results in providing efficient remedy against secondary caries. CuO-Chitosan nanoparticles reinforced dental adhesive discs cause significant upsurge in reduction of Lactobacillus acidophillus and Streptococcus mutans. Also, the augmentation of mechanical properties, water sorption and solubility plus slow and sustained release profile and slight variation of shear bond strength is attained. Taken together, the chemically synthesized CuO nanoparticles and CuO-Chitosan nanoparticles have proven to be promising candidates having enormous potential to be utilized in drug delivery and nanotheranostics.

Antibacterial activity and debonding force of different lingual retainers bonded with conventional composite and nanoparticle containing composite: An in vitro study.

BACKGROUND: To evaluate the antibacterial activity and debonding force of retainers bonded with conventional and nanoparticle (TiO2) containing composite. METHODOLOGY: Antibacterial activity was tested against Streptococcus mutans and Lactobacillus acidophilus using disk agar diffusion, biofilm inhibition, and eluted components tests. For the eluted components test, colony counts of bacteria were tested on 0, 3, and 30 days. Three different retainers were bonded to the lingual surface of extracted lower incisors using conventional and 1% TiO2 composite. Samples were divided as follows: Group 1: 1a, stainless steel retainer (Bond-a-Braid) with conventional composite, and 1b, stainless steel retainer with nanoparticle composite; Group 2: 2a, titanium retainer with conventional composite, and 2b, titanium retainer with nanoparticle composite; Group 3: 3a, fiber-reinforced retainer (Interlig) with conventional composite, and 3b, fiber-reinforced retainer with nanoparticle composite. The Instron stereomicroscope was used to test debonding force and failure sites respectively. RESULTS: In the disk agar diffusion test, TiO2 composite has shown more inhibition zones. Biofilm inhibition test showed a significant decrease in colony counts of both organisms in the TiO2 group. The eluted component test showed a significant decrease in colony counts from day 0 to day 30 in the TiO2 group compared with the control group. The highest debonding force was observed in stainless steel retainers with conventional composite, and lowest in fiber-reinforced composite retainers with TiO2 composite, with no significant difference in Adhesive Remnant Index scores. CONCLUSION: The TiO2 composite group showed greater antibacterial activity without compromising the bond strength, which was statistically significant. Compared with other groups, stainless steel wires bonded with conventional composite showed the highest debonding force.

A quaternary ammonium silane antimicrobial triggers bacterial membrane and biofilm destruction.

To study the antimicrobial effects of quaternary ammonium silane (QAS) exposure on Streptococcus mutans and Lactobacillus acidophilus bacterial biofilms at different concentrations. Streptococcus mutans and Lactobacillus acidophilus biofilms were cultured on dentine disks, and incubated for bacterial adhesion for 3-days. Disks were treated with disinfectant (experimental QAS or control) and returned to culture for four days. Small-molecule drug discovery-suite was used to analyze QAS/Sortase-A active site. Cleavage of a synthetic fluorescent peptide substrate, was used to analyze inhibition of Sortase-A. Raman spectroscopy was performed and biofilms stained for confocal laser scanning microscopy (CLSM). Dentine disks that contained treated dual-species biofilms were examined using scanning electron microscopy (SEM). Analysis of DAPI within biofilms was performed using CLSM. Fatty acids in bacterial membranes were assessed with succinic-dehydrogenase assay along with time-kill assay. Sortase-A protein underwent conformational change due to QAS molecule during simulation, showing fluctuating alpha and beta strands. Spectroscopy revealed low carbohydrate intensities in 1% and 2% QAS. SEM images demonstrated absence of bacterial colonies after treatment. DAPI staining decreased with 1% QAS (p < 0.05). Fatty acid compositions of dual specie biofilm increased in both 1% and 2% QAS specimens (p < 0.05). Quaternary ammonium silane demonstrated to be a potent antibacterial cavity disinfectant and a plaque inhibitor and can be of potential significance in eliminating caries-forming bacteria.

Evodiamine has therapeutic efficacy in ulcerative colitis by increasing Lactobacillus acidophilus levels and acetate production.

Emerging evidence implicates gut microbiota have an important role in ulcerative colitis (UC). Previous study indicated that Evodiamine (EVO) can alleviate colitis through downregulating inflammatory pathways. However, specific relationship between EVO-treated colitis relief and regulation of gut microbiota is still unclear. Here, our goal was to determine the potential role of gut microbiota in the relief of UC by EVO. By using pathology-related indicators, 16S rRNA sequencing and metabolomics profiling, we assessed the pharmacological effect of EVO on dextran sulfate sodium (DSS)-induced colitis rats as well as on the change of gut microbiota and metabolism. Fecal derived from EVO-treated rats was transplanted into colitis rats to verify the effect of EVO on gut microbiota, and 'driver bacteria' was found and validated by 16S rRNA sequencing, metagenome and qRT-PCR. The effect of Lactobacillus acidophilus (L. acidophilus) was investigated by vivo experiment, microbiota analysis, Short-chain fatty acids (SCFAs) quantification and colon transcriptomics. EVO reduced the susceptibility to DSS-induced destruction of epithelial integrity and severe inflammatory response, and regulated the gut microbiota and metabolites. Fecal Microbiota Transplantation (FMT) alleviated DSS-induced colitis, increased the abundance of L. acidophilus and the level of acetate. Furthermore, gavaged with L. acidophilus reduced pro-inflammatory cytokines, promoted the increase of goblet cells and the secretion of antimicrobial peptides, regulated the ratio of Firmicutes/Bacteroidetes and increased the level of acetate. Our results indicated that EVO mitigation of DSS-induced colitis is associated with increased in L. acidophilus and protective acetate production, which may be a promising strategy for treating UC.

Influence of freeze-drying and spray-drying preservation methods on survivability rate of different types of protectants encapsulated Lactobacillus acidophilus FTDC 3081.

The aims of this study were to compare the effectiveness of different drying methods and to investigate the effects of adding a series of individual protectant such as skim milk, sucrose, maltodextrin, and corn starch for preserving Lactobacillus acidophilus FTDC 3081 cells during spray and freeze-drying and storage at different temperatures. Results showed a remarkable high survival rate of 70-80% immediately after spray- and freeze-drying in which the cell viability retained at the range of 109 to 1010 CFU/mL. After a month of storage, maltodextrin showed higher protective ability on both spray- and freeze-dried cells as compared to other protective agents at 4°C, 25°C, and 40°C. A complete loss in viability of spray-dried L. acidophilus FTDC 3081 was observed after a month at 40°C in the absence of protective agent.

Evaluation of pyroligneous acid as a therapeutic agent against Salmonella in a simulated gastrointestinal tract of poultry.

Pyroligneous acid (PA) was evaluated as a potential alternative to therapeutic antibiotics in poultry. Antimicrobial activity of PA was studied at acidic pH (2.0) and neutral pH (7.0) of the liquid against Salmonella enterica and Lactobacillus acidophilus. Acidic PA gave a MIC value of 0.8% (v/v) and 1.6% (v/v), and neutralized PA gave a MIC value of 1.6% (v/v) and 3.2% (v/v) against S. enterica and L. acidophilus respectively. Acidic PA was evaluated at different concentrations in a simulated poultry digestive tract and cecal fermentation to study its effect on the cecal microflora and fermentation profile. PA at a concentration of 1.6% (v/v) completely inhibited S. enterica and was also found to have a similar effect on lactobacilli count as compared with the control (p = 0.17). Additionally, PA at this concentration was found not to have a significant effect on acetic acid production after 24 h of cecal fermentation (p = 0.20). Graphical abstract.

Is Acrylamide as Harmful as We Think? A New Look at the Impact of Acrylamide on the Viability of Beneficial Intestinal Bacteria of the Genus Lactobacillus.

The impact of acrylamide (AA) on microorganisms is still not clearly understood as AA has not induced mutations in bacteria, but its epoxide analog has been reported to be mutagenic in Salmonella strains. The aim of the study was to evaluate whether AA could influence the growth and viability of beneficial intestinal bacteria. The impact of AA at concentrations of 0-100 µg/mL on lactic acid bacteria (LAB) was examined. Bacterial growth was evaluated by the culture method, while the percentage of alive, injured, and dead bacteria was assessed by flow cytometry after 24 h and 48 h of incubation. We demonstrated that acrylamide could influence the viability of the LAB, but its impact depended on both the AA concentration and the bacterial species. The viability of probiotic strain Lactobacillus acidophilus LA-5 increased while that of Lactobacillus plantarum decreased; Lactobacillus brevis was less sensitive. Moreover, AA influenced the morphology of L. plantarum, probably by blocking cell separation during division. We concluded that acrylamide present in food could modulate the viability of LAB and, therefore, could influence their activity in food products or, after colonization, in the human intestine.

Enhanced Acid Tolerance in Lactobacillus acidophilus by Atmospheric and Room Temperature Plasma (ARTP) Coupled with Adaptive Laboratory Evolution (ALE).

The aim of this study was to improve the acid tolerance of Lactobacillus acidophilus by combining atmospheric and room temperature plasma (ARTP) mutation with adaptive laboratory evolution (ALE). To achieve a high mutation efficiency, 60 s was determined as the ideal exposure time for ARTP mutation of L. acidophilus with a survival rate of 5.91%. The ARTP-ALE mutant strain LAartp-ale2 displayed increased lactic acid stress tolerance with survival rates of 75.67% and 25.78% when cultured in pH 3.0 and 2.5, respectively, for 3 h. Physiological analysis revealed that the ARTP-ALE mutant exhibited a lower inner membrane permeability than that of the parental strain during acid stress. Furthermore, the mutant LAartp-ale2 produced more biofilm in response to lactic acid-induced acid stress and showed an increased hydrophobicity (87.2%) when compared to the parent strain (76.2%) at pH 2.5. LAartp-ale2 exhibited a higher unsaturated fatty acid (UFA) to saturated fatty acid (SFA) ratio that affected the physical state of the cell membrane for increased survival in pH 3.0 and 2.5. The mutation with ARTP coupled with ALE in the present study proved to be effective in enhancing the acid tolerance of L. acidophilus for potential industrial use.

Prebiotic activity of monofloral honeys produced by stingless bees in the semi-arid region of Brazilian Northeastern toward Lactobacillus acidophilus LA-05 and Bifidobacterium lactis BB-12.

This study assessed the in vitro prebiotic effects of honeys from Ziziphus joazeiro Mart. (juazeiro; J) and Mimosa arenosa Willd Poir (jurema branca; JB) produced by native stingless bees, namely Melipona subnitida Ducke (jandaíra; J) and M. scutellaris Latrelle (uruçu; U), in the Brazilian Northeastern semi-arid region toward the probiotics Lactobacillus acidophilus LA-05 and Bifidobacterium animalis subsp. lactis BB-12. Cells of the probiotic strains were enumerated over 48 h of cultivation in broths containing each honey (JJ, JU, JBJ or JBU) as a sole carbon source. The metabolic activities of probiotic strains in these media were assessed by measuring changes in pH values and sugars, organic acids and phenolics contents. All honeys (20 or 30 g/L) exerted growth promoting effects and displayed positive prebiotic activity scores (0.94-1.22) on tested probiotics. JJ showed the highest (p < 0.05) stimulatory effects on probiotics growth and prebiotic scores. At the end of the cultivation period, counts of L. acidophilus LA-05 and B. lactis BB-12 increased (p < 0.05) more than 2 log in broths regardless the monofloral honey added. The pH values and sugars contents decreased (p < 0.05), while the organic acids contents increased (p < 0.05) during cultivation of probiotics in broths containing JJ, JU, JBJ or JBU as carbon source. After 48 h of cultivation, contents of gallic, caftaric and caffeic acid, catechin and procyanidins (B1 and B2) decreased (p < 0.05) in media containing JJ, JU, JBJ or JBU despite of the inoculated probiotic. JJ honey presented overall the better stimulatory effects on the growth and metabolism of L. acidophilus LA-05 and B. lactis BB-12. These results showed for the first time the potential prebiotic properties of four monofloral honeys produced by stingless bees in the Brazilian Northeastern semi-arid region.