Lacticaseibacillus Rhamnosus GG Research Articles

Assessing the efficacy of probiotics in augmenting bovine reproductive health: an integrated in vitro, in silico, and in vivo study.

The aim of this study was to isolate and characterize bovine-vaginal probiotics genotypically and phenotypically using in silico and evaluate their in vivo performance in buffaloes with endometritis. For the in vitro isolation and characterization, vaginal swabs were collected from 34 cows and 17 buffaloes, and 709 primary bacterial isolates with probiotic activity were obtained using MRS agar media. Two isolates Lactiplantibacillus plantarum KUGBRC (LPKUGBRC) and Pediococcus pentosaceus GBRCKU (PPGBRCKU) demonstrated optimum in vitro probiotic activities as compared to Lacticaseibacillus rhamnosus GG including, acid production, secretion of fatty acids and exopolysaccharide, cell surface hydrophobicity, self-aggregating and co-aggregating capacity with pathogens, anti-microbial activity and bacteriocin-like compounds against pathogens Escherichia coli and Staphylococcus aureus in cell-free supernatant and absence of hemolytic activity. Their phenotypic capacity was confirmed by analyzing the whole genome sequencing data and identifying genes and pathways associated with probiotic properties. These probiotic isolates have shown no virulence genes were discovered in their genomic study. In vivo study of 92 buffaloes suffering from clinical endometritis with purulent cervico-vaginal mucus (CVM) were randomly allocated 40 × 108 CFU/ml LPKUGBRC and PPGBRCKU and 40 ml Normal saline. The LPKUGBRC reduced the duration between administration of probiotic to induction of healthy estrus significantly. However, no effect was observed on pregnancy rate. These results suggest that LPKUGBRC and PPGBRCKU probiotic bacteria demonstrate probiotic efficiency and adaptability. Further sourced from the same niche as the targeted infection, they offer a distinct advantage in targeting the specific microbial population associated with endometritis. The findings of this study highlight the potential of LPKUGBRC and PPGBRCKU probiotics in treating endometritis and suggest further exploration of their clinical applications.

Limnospira indica PCC8005 and Lacticaseibacillus rhamnosus GG Mixed Dietary Combination Reduces Pelvic Irradiation-Induced Symptoms in Mice

Throughout their cancer treatments, around half of all patients will undergo irradiation that is accompanied by several side effects reducing their quality of life and leading to the interruption or extension of their treatment course. Pelvic irradiation leads to the triggering of mucositis and dysbiosis, further impairing the daily life of the patients. In this work, we address the ability of Limnospira indica strain PCC 8005 and Lacticaseibacillus rhamnosus GG ATCC 53103 in alleviating the above-mentioned side effects triggered by the local pelvic irradiation of 12 Gy in a mouse model. We found that the combinatorial administration of these food supplements was able to confer partial tight junction protection while the bacterial translocation towards the mesenteric lymph nodes was found to be identical between the saline sham-irradiated and supplemented irradiated group. Furthermore, the supplemented group did not present a significant shift in microbial composition following pelvic irradiation, indicating that the bacterial formulation was able to mitigate the dysbiosis induced by the latter treatment, as observed in the saline irradiated group. These very promising results will be further completed by investigating the mode of action and/or active molecules mediating the beneficial effects of both L. indica PCC 8005 and L. rhamnosus GG.

Probiotic Properties and Antioxidant Activity In Vitro of Lactic Acid Bacteria.

The properties of probiotics such as lactic acid bacteria (LAB) have been widely studied over the last decades. In the present study, four different LAB species, namely Lactobacillus gasseri ATCC 33323, Lacticaseibacillus rhamnosus GG ATCC 53103, Levilactobacillus brevis ATCC 8287, and Lactiplantibacillus plantarum ATCC 14917, were investigated in order to determine their ability to survive in the human gut. They were evaluated based on their tolerance to acids, resistance to simulated gastrointestinal conditions, antibiotic resistance, and the identification of genes encoding bacteriocin production. All four tested strains demonstrated high resistance to simulated gastric juice after 3 h, and the viable counts revealed declines in cell concentrations of less than 1 log cycle. L. plantarum showed the highest level of survival in the human gut, with counts of 7.09 log CFU/mL. For the species L. rhamnosus and L. brevis, the values were 6.97 and 6.52, respectively. L. gasseri, after 12 h, showed a 3.96 log cycle drop in viable counts. None of the evaluated strains inhibited resistance to ampicillin, gentamicin, kanamycin, streptomycin, erythromycin, clindamycin, tetracycline, or chloramphenicol. With regard to bacteriocin genes, the Pediocin PA gene was identified in Lactiplantibacillus plantarum ATCC 14917, Lacticaseibacillus rhamnosus GG ATCC 53103, and Lactobacillus gasseri ATCC 33323. The PlnEF gene was detected in Lactiplantibacillus plantarum ATCC 14917 and Lacticaseibacillus rhamnosus GG ATCC 53103. The Brevicin 174A and PlnA genes were not detected in any bacteria. Moreover, the potential antioxidant activity of LAB's metabolites was evaluated. At the same time, the possible antioxidant activity of metabolites of LAB was first tested using the free radical DDPH• (a, a-Diphenyl-β-Picrylhydrazyl) and then evaluated with regard to their radical scavenging activity and inhibition against peroxyl radical induced DNA scission. All strains showed antioxidant activity; however, the best antioxidant activity was achieved by L. brevis (94.47%) and L. gasseri (91.29%) at 210 min. This study provides a comprehensive approach to the action of these LAB and their use in the food industry.

Fecal virome transfer improves proliferation of commensal gut Akkermansia muciniphila and unexpectedly enhances the fertility rate in laboratory mice

ABSTRACT Probiotics are intended to improve gastrointestinal health when consumed. However, the probiotics marketed today only colonize the densely populated gut to a limited extent. Bacteriophages comprise the majority of viruses in the human gut virome and there are strong indications that they play important roles in shaping the gut microbiome. Here, we investigate the use of fecal virome transplantation (FVT, sterile filtrated feces) as a mean to alter the gut microbiome composition to lead the way for persistent colonization of two types of probiotics: Lacticaseibacillus rhamnosus GG (LGG) representing a well-established probiotic and Akkermansia muciniphila (AKM) representing a putative next-generation probiotic. Male and female C57BL/6NTac mice were cohoused in pairs from 4 weeks of age and received the following treatment by oral gavage at week 5 and 6: AKM+FVT, LGG+FVT, probiotic sham (Pro-sham)+FVT, LGG+Saline, AKM+Saline, and control (Pro-sham+Saline). The FVT donor material originated from mice with high relative abundance of A. muciniphila. All animals were terminated at age 9 weeks. The FVT treatment did not increase the relative abundance of the administered LGG or AKM in the recipient mice. Instead FVT significantly (p < 0.05) increased the abundance of naturally occurring A. muciniphila compared to the control. This highlights the potential of propagating the existing commensal “probiotics” that have already permanently colonized the gut. Being co-housed male and female, a fraction of the female mice became pregnant. Unexpectedly, the FVT treated mice were found to have a significantly (p < 0.05) higher fertility rate independent of probiotic administration. These preliminary observations urge for follow-up studies investigating interactions between the gut microbiome and fertility.

Membrane Emulsification as an Emerging Method for Lacticaseibacillus rhamnosus GG® Encapsulation.

The online version contains supplementary material available at 10.1007/s11947-023-03099-w.

Online Monitoring of the Growth of Probiotic Bacteria and Metabolites in the Fermentation of a Teff Substrate Using Model-Based Calibration of 2D Fluorescence Spectra.

The demand for probiotic bacteria-fermented food products is increasing; however, the monitoring of the fermentation process is still challenging when using conventional approaches. A classical approach requires a large amount of offline data to calibrate a chemometric model using fluorescence spectra. Fluorescence spectra provide a wide range of online information during the process of cultivation, but they require a large amount of offline data (which involves laborious work) for the calibration procedure when using a classical approach. In this study, an alternative model-based calibration approach was used to predict biomass (the growth of Lactiplantibacillus plantarum A6 (LPA6) and Lacticaseibacillus rhamnosus GG (LCGG)), glucose, and lactic acid during the fermentation process of a teff-based substrate inoculated with mixed strains of LPA6 and LCGG. A classical approach was also applied and compared to the model-based calibration approach. In the model-based calibration approach, two-dimensional (2D) fluorescence spectra and offline substituted simulated data were used to generate a chemometric model. The optimum microbial specific growth rate and chemometric model parameters were obtained simultaneously using a particle swarm optimization algorithm. The prediction errors for biomass, glucose, and lactic acid concentrations were measured between 6.1 and 10.5%; the minimum error value was related to the prediction of biomass and the maximum one was related to the prediction of glucose using the model-based calibration approach. The model-based calibration approach and the classical approach showed similar results. In conclusion, the findings showed that a model-based calibration approach could be used to monitor the process state variables (i.e., biomass, glucose, and lactic acid) online in the fermentation process of a teff-based substrate inoculated with mixed strains of LPA6 and LCGG. However, glucose prediction showed a high error value.

Effects of fermenting with Lacticaseibacillus rhamnosus GG on quality attributes and storage stability of buffalo milk yogurt incorporated with bael (Aegle marmelos) fruit pulp

BackgroundProducing functional food by adding fruits or fruit pulps have attracted great attention. Simultaneously, buffalo milk is gaining an increasing demand as an alternative to cow milk. Thus, value addition and diversification of buffalo milk products have gained much commercial and research interest. Hence, we aimed to investigate the potentials of developing and characterizing probiotic enriched buffalo milk yogurts with bael fruit pulp using exopolysaccharides producing probiotic Lacticaseibacillus rhamnosus GG (LGG). MethodsFour types of buffalo milk yogurts were tested, e.g. fermenting with the yogurt starter culture only (e.g., control) and fermenting with the combination of yogurt starter culture and LGG with varying levels (w/v) of bael fruit pulp incorporations, i.e., 0%, 5% and 10%. Variation in pH, syneresis, hardness, probiotic viability and sensory attributes during 21 days of storage in 4 °C were assessed for all treatments. ResultsFermenting with LGG had a positive effect on post-acidification and syneresis rate compared to the control yogurt. Bael incorporation did not affect the post-acidification, but significantly decreased the level of syneresis at the end of storage. All probiotic formulations maintained LGG counts of >107 CFU/mL and the highest counts were observed in 5% (w/v) bael incorporated yogurt. ConclusionsResults confirmed the possibility of using buffalo milk yogurt as an ideal matrix to deliver LGG with promising probiotic capacity. The use of 5% bael incorporation provides an optimal combination for synbiotic product development.

Alginate encapsulation improves probiotics survival in carbonated sodas and beers.

Probiotic functionalization of non-dairy beverages has been garnering interest to provide dairy-sensitive populations with greater probiotic product varieties. The addition of probiotics into popularly consumed beverages-carbonated sodas and beers, presents an interesting challenge as the presence of acidic pH, hops-derived compounds, and ethanol have highly deleterious effects. Herein, alginate encapsulation was proposed to improve probiotics viability within sodas and beers. Three probiotics, namely Lacticaseibacillus rhamnosus GG, Escherichia coli Nissle 1917, and Bifidobacterium longum were encapsulated in alginate spheres and exposed to Coca-Cola, 7-Up, Tiger Beer, and Guinness under refrigerated, room temperature and simulated gastric fluid conditions. Results demonstrate that alginate encapsulation significantly improved the viabilities of all three probiotics in various beverages and conditions. Refrigerated storage better preserved probiotic viabilities and reduced the formation of the probiotic metabolic by-product, L-lactate, than at room temperature storage. Findings here could provide beverage manufacturers with a novel way to develop probiotic-sodas and probiotic-beers through encapsulation.

Lactobacillus Bacteremia and Probiotics: A Review.

Lactobacilli are widely found in nature, are commensal microbes in humans, and are commonly used as probiotics. Concerns about probiotic safety have arisen due to reports of bacteremia and other Lactobacillus-associated infections. We reviewed the literature for articles on the pathogenicity of Lactobacillus spp. bacteremia and reports of probiotics in these patients. Our aim is to review these articles and update the present knowledge on the epidemiology of Lactobacillus spp. bacteremia and determine the role of probiotics in Lactobacillus bacteremia. Lactobacillus bacteremia is infrequent but has a higher risk of mortality and risk factors, including severe underlying diseases, immune system suppression, admission to intensive care units, and use of central venous catheters. A variety of Lactobacillus species may cause bacteremia and may or may not be associated with probiotic exposure. To determine if oral probiotics are the source of these infections, the blood isolates and the oral probiotic strain(s) must be compared by sensitive identification methods. The prevalence of Lactobacillus bacteremia is infrequent but is more common in patients taking probiotics compared to those not taking probiotics. Three probiotics (Lacticaseibacillus rhamnosus GG, Lactiplantibacillus plantarum, and Lacticaseibacillus paracasei) were directly linked with blood isolates from bacteremia patients using molecular identification assays.

Milk protein-based cryogel monoliths as novel encapsulants of probiotic bacteria. Part I: Microstructural, physicochemical, and mechanical characterisation

The implementation of cryogels as alternative xero-carriers for embedding labile bioactive compounds including probiotic living cells is becoming more popular. In the present work, milk protein-based cryogels were developed by freeze-drying of indirectly acidified protein gels (10% wt. in sodium caseinate or whey protein isolate blended at varying mass ratios i.e., 1:0, 3:1, 1:1, 1:3 and 0:1) comprising Lacticaseibacillus rhamnosus GG (LGG) living cells, trehalose (5% wt.), glucose (1% wt.) and glycerol (2.5% wt.). The physicochemical, microstructural, and mechanical characteristics of the cryogels conveying LGG were measured. The acid gel contraction during the fermentation notably influenced the macroscopic (volume contraction) and microscopic aspects (porosity, thickness, and uniformity of the wall material construct) of the cryogels. The amount of monolayer water content (11.5−14.0 g.100 g−1), and total surface area (403−488 m2 g−1) as well as the glass transition temperature (Tg = 33−46 °C), of the cryogels increased proportionally to the whey protein content. X-ray microtomography analysis revealed that the mixed protein cryogels were characterised by lower porosities than their individual protein exemplars. However, the uniformity and the structure conformation of the protein network constructs were altered according to the milk protein composition of the cryogels. All the cryogels exhibited a brittle and plastic behaviour when subjected to indentation forces, with the hardest and stiffest cryogels exerting the lowest water reconstituting capacity. The microstructural assessment of the wall material at nanoscale level evidenced a satisfactory intrenching of the LGG cells, which substantiates the feasibility of the cryogels as novel probiotic xero-scaffolds.

Limnospira indica PCC 8005 Supplementation Prevents Pelvic Irradiation-Induced Dysbiosis but Not Acute Inflammation in Mice.

Pelvic irradiation-induced mucositis secondarily leads to dysbiosis, which seriously affects patients' quality of life after treatment. No safe and effective radioprotector or mitigator has yet been approved for clinical therapy. Here, we investigated the potential protective effects of fresh biomass of Limnospira indica PCC 8005 against ionizing irradiation-induced mucositis and dysbiosis in respect to benchmark probiotic Lacticaseibacillus rhamnosus GG ATCC 53103. For this, mice were supplemented daily before and after 12 Gy X-irradiation of the pelvis. Upon sacrifice, food supplements' efficacy was assessed for intestinal barrier protection, immunomodulation and changes in the microbiota composition. While both could not confer barrier protection or significant immunomodulatory effects, 16S microbial profiling revealed that L. indica PCC 8005 and L. rhamnosus GG could prevent pelvic irradiation-induced dysbiosis. Altogether, our data show that-besides benchmarked L. rhamnosus GG-L. indica PCC 8005 is an interesting candidate to further explore as a radiomitigator counteracting pelvic irradiation-induced dysbiosis in the presented in vivo irradiation-gut-microbiota platform.

Comparative Genomic Analysis and Physiological Properties of Limosilactobacillus fermentum SMFM2017-NK2 with Ability to Inflammatory Bowel Disease.

The objective of this study was to evaluate the anti-inflammatory effect of Latilactobacillus sakei SMFM2017-NK1 (LS1), L. sakei SMFM2017-NK3 (LS2), and Limosilactobacillus fermentum SMFM2017-NK2 (LF) on colitis using an animal model. DSS (dextran sulfate sodium salt) was orally injected into C57BL/6N mice to induce inflammation in the colon for seven days. Colitis mice models were treated with LS1, LS2, and LF, respectively, and Lacticaseibacillus rhamnosus GG (LGG) was used as a positive control. During oral administration of lactic acid bacteria, the weights of the mice were measured, and the disease activity index (DAI) score was determined by judging the degree of diarrhea and bloody stool. When comparing the differences between the minimum weight after DSS administration and the maximum weight after lactic acid bacteriaadministration were compared, the LF-treated group showed the highest weight gain at 8.91%. The DAI scores of the LF, LS2, and LGG groups were lower than that of the control group. After sacrifice, mRNA expression levels for proinflammatory cytokines (TNF-α, IL-1β, IL-6, and IFN-γ) and mediators (iNOS and COX-2) in the colon were measured. LF was selected as a superior strain for anti-inflammation in the colon. It was further analyzed to determine its biochemical characteristics, cytotoxicity, and thermal stability. Catalase and oxidase activities for LF were negative. In cytotoxicity and heat stability tests, the LF group had higher cell viability than the LGG group. The genome of LF was obtained, and 5682 CDS, 114 tRNA, 2 RNA, and 5 repeat regions were predicted. Especially, LF could be distinguished from the other three L. fermentum strains based on taxonomic profiling, specific orthologous genes of the strain, and genomic variants. The results of this study suggest that L. fermentum SMFM2017-NK2 is a novel strain with an anti-inflammatory effect on colitis.

Role of N-Terminal Extensional Long α-Helix in the Arylesterase from Lacticaseibacillus rhamnosus GG on Catalysis and Stability

In the α/β hydrolases superfamily, the extra module modulated enzymatic activity, substrate specificity, and stability. The functional role of N-terminal extensional long α-helix (Ala2-Glu29, designated as NEL-helix) acting as the extra module in the arylesterase LggEst from Lacticaseibacillus rhamnosus GG had been systemically investigated by deletion mutagenesis, biochemical characterization, and biophysical methods. The deletion of the NEL-helix did not change the overall structure of this arylesterase. The deletion of the NEL-helix led to the shifting of optimal pH into the acidity and the loss of thermophilic activity. The deletion of the NEL-helix produced a 10.6-fold drop in catalytic activity towards the best substrate pNPC10. NEL-Helix was crucial for the thermostability, chemical resistance, and organic solvents tolerance. The deletion of the NEL-helix did not change the overall rigidity of enzyme structure and only reduced the local rigidity of the active site. Sodium deoxycholate might partially replenish the loss of activity caused by the deletion of the NEL-helix. Our research further enriched the functional role of the extra module on catalysis and stability in the α/β hydrolase fold superfamily.

Impact of Pomegranate on Probiotic Growth, Viability, Transcriptome and Metabolism

Despite rising interest in understanding intestinal bacterial survival in situ, relatively little attention has been devoted to deciphering the interaction between bacteria and functional food ingredients. Here, we examined the interplay between diverse beneficial Lactobacillaceae species and a pomegranate (POM) extract and determined the impact of this functional ingredient on bacterial growth, cell survival, transcription and target metabolite genesis. Three commercially available probiotic strains (Lactobacillus acidophilus NCFM, Lacticaseibacillus rhamnosus GG and Lactiplantibacillus plantarum Lp-115) were used in growth assays and flow cytometry analysis, indicating differential responses to the presence of POM extract across the three strains. The inclusion of POM extract in the growth medium had the greatest impact on L. acidophilus cell counts. LIVE/DEAD staining determined significantly fewer dead cells when L. acidophilus was grown with POM extract compared to the control with no POM (1.23% versus 7.23%). Whole-transcriptome analysis following exposure to POM extract showed markedly different global transcriptome responses, with 15.88% of the L. acidophilus transcriptome, 19.32% of the L. rhamnosus transcriptome and only 2.37% of the L. plantarum transcriptome differentially expressed. We also noted strain-dependent metabolite concentrations in the medium with POM extract compared to the control medium for punicalagin, ellagic acid and gallic acid. Overall, the results show that POM extract triggers species-specific responses by probiotic strains and substantiates the rising interest in using POM as a prebiotic compound.

Synergistic Inhibitory Effect of Lactobacillus Cell Lysates and Butyrate on Poly I:C-Induced IL-8 Production in Human Intestinal Epithelial Cells.

Postbiotics include cell lysates (CLs), enzymes, cell wall fragments, and heat-killed bacteria derived from probiotics. Although postbiotics are increasingly being considered for their potential health-promoting properties, the effects of postbiotics on virus-mediated inflammatory responses in the intestine have not been elucidated. Hence, the present study aimed to examine whether CLs of Lactipantibacillus plantarum (LP CL) and Lacticaseibacillus rhamnosus GG (LR CL) could inhibit virus-mediated inflammatory responses in the human intestinal epithelial cell line HT-29 in vitro. Pretreatment with LP CL and LR CL significantly inhibited interleukin (IL)-8 production, which was induced by poly I:C, a synthetic analog of double-stranded RNA (dsRNA) viruses, at the mRNA and protein levels in HT-29 cells. However, peptidoglycans and heat-killed L. plantarum and L. rhamnosus GG did not effectively inhibit IL-8 production. LP CL and LR CL attenuated the poly I:C-induced phosphorylation of ERK and JNK and the activation of NF-κB, suggesting that these CLs could inhibit poly I:C-induced IL-8 production by regulating intracellular signaling pathways in HT-29 cells. Furthermore, among the short-chain fatty acids, butyrate enhanced the inhibitory effect of CLs on poly I:C-induced IL-8 production at the mRNA and protein levels in HT-29 cells, while acetate and propionate did not. Taken together, these results suggest that both LP CL and LR CL could act as potent effector molecules that can inhibit virus-mediated inflammatory responses and confer synergistic inhibitory effects with butyrate in human intestinal epithelial cells.

Microencapsulation of Lacticaseibacillus rhamnosus GG and L. plantarum 299V by reverse spherification: a promising method to improve the survival of probiotics.

This research reports the first application of the reverse spherification (RVS) method for encapsulation of two probiotics (Lacticaseibacillus rhamnosus GG and L. plantarum 299V) compared to the basic spherification (BS). These probiotics were encapsulated in different solutions encompassing various contents of alginate, gelatin, and gellan gum. The RVS bead diameters was about 1.5 times bigger and hardness was 70%-80% lower than BS samples. As determined by Raman spectral mapping, the RVS beads had two calcium alginate walls but the BS beads had only one. The inner wall of the RVS beads was more than three times thicker than outer wall. The encapsulation yields of gelatin/gellan gum and gelatin beads prepared by both methods were >1.5% alginate beads. All the RVS-prepared beads were resistant to stomach acid and showed no significant reduction in the intestine. Furthermore, the incorporation of gelatin and gellan gum into alginate led to higher cell protection. For 1.5% alginate beads, <67% survival was achieved after acid exposure but in others, >77% survival was observed; RVS beads were about 1 log above than BS ones. The proposed novel microencapsulation method efficiently increased the viability of probiotic bacteria compared to the conventional approaches.

Viability and resistance of Lacticaseibacillus rhamnosus GG to passion fruit beverages with whey protein isolate

This study characterized the beverage of passion fruit and whey protein added with Lacticaseibacillus rhamnosus GG (LGG). The acidity, pH, soluble solids, protein, phenolic compounds, and color of the sample did not differ during the 28 days at 5°C. The LGG counts in the beverage remained >7.70 log CFU/mL during the evaluation period. However, the LGG count was significantly reduced after the in vitro simulated gastrointestinal tract (GIT) evaluation (reduction of 5 logarithmic cycles after processing). On the other hand, the LGG resistance to GIT increased during the shelf-life of the beverage (LGG count of ~ 4.0 log CFU/mL after 28 days). In addition, the LGG count after GIT increased by approximately 2.0 logarithmic cycles when microencapsulation and fermentation technologies were used in beverage processing. Thus, the product has the potential to be commercialized, due to high viability of LGG use to increase the resistance of LGG to GIT.

Антагонистическая активность пробиотиков при кампилобактериозе

Campylobacter species are globally recognized as the most common causative agents of acute intestinal infections. In recent years, Campylobacter spp. have become increasingly resistant to antibiotics, necessitating the search for new methods of treatment. Objective. To evaluate the antagonistic activity of different probiotics and autoprobiotic enterococci against Campylobacter spp. Materials and methods. Clinical isolates of Campylobacter coli and Campylobacter jejuni were obtained from the microbial culture collection in the Pediatric Research and Clinical Center for Infectious Diseases, Saint Petersburg, Russian Federation. The collection included cryopreserved samples of Campylobacter spp. strains isolated from stool specimens of children with campylobacteriosis. Lactobacillus rhamnosus ATCC 53103, Bifidobacterium longum PXN 30, Enterococcus faecium L3, Lactobacillus acidophilus, Sacharomyces boulardii, Limosilactobacillus reuteri DSM 17938, Lacticaseibacillus rhamnosus GG, Lactobacillus plantarum 8R-A3, Bifidobacterium animalis subsp. lactis (BB-12) at the concentration of 108 CFU/mL were used as probiotic strains. Autoprobiotic (indigenous) strains of E. faecium at the concentration of 108 CFU/mL isolated from stool specimens of children with campylobacteriosis were used as autoprobiotics. Results. No strain of Campylobacter spp. was found to be resistant to the probiotics and autoprobiotics included in the study. Only one strain of C. jejuni 31 was susceptible to all probiotic and autoprobiotic strains. B. animalis subsp. lactis BB12 strains at the concentrations of 109 and 106 CFU showed the highest anti-Campylobacter activity, inhibiting the growth of 36 (85.71%) and 38 (90.48%) isolated Campylobacter species, respectively. High anti-Campylobacter activities were also seen in L. rhamnosus GG and L. reuteri DSM 17938, which inhibited the growth of 29 (69.05%) and 31 (73.81%), 28 (66.67%) and 29 (69.05%) clinical Campylobacter isolates, respectively. In terms of the effectiveness in inhibiting Campylobacter spp., autoprobiotics did not significantly differ from probiotic strains of B. longum + and/or E. faecium SF68, preventing the growth of 35–45% of the studied Campylobacter spp. strains. Conclusion. The use of specially selected probiotic and autoprobiotic strains by in vitro test in campylobacteriosis may contribute to the rapid elimination of pathogens due to the ability of some strains to inhibit the growth of Campylobacter spp. Key words: Campylobacter, probiotics, autoprobiotics, anti-Campylobacter activity

Impact of electro-activated whey on growth, acid and bile resistance of Lacticaseibacillus rhamnosus GG and Lactobacillus acidophilus ATCC 4356

The aim of this work was to study the impact of electro-activated whey containing lactulose on the growth and survival Lacticaseibacillus rhamnosus GG and Lactobacillus acidophilus ATCC 4356 in acidic and bile salts containing media. Electro-activated whey was compared to whey and MRS alone and supplemented with lactulose. The results showed that OD600 was the highest for all these bacteria when grown in the electro-activated medium. At the same time, the obtained results showed that the initial growth phase was the most delayed in this medium. The OD600 results were verified by the bacteria plating method on nutrient agar. The obtained data showed that for each given bacteria, no significant difference was observed according to the CFU/mL results. Thus, it has been suggested that electro-activated whey could have a significant effect of bacterial fitness by enhancing their activity even at an equivalent population in each medium. A study of the stability of the probiotic bacteria for 14 days refrigerated storage at pH 4.6 and in the presence of bile salts revealed that the growth substrate did not significantly affect bacterial survival during this storage period and that all the tested probiotic bacteria remained close to 109 CFU/mL. The 16S rRNA gene sequencing of Lacticaseibacillus rhamnosus GG after 24 h growth in different media showed highly significant difference in upregulated and downregulated genes between the electro-activated whey and the regular sweet whey even when it was supplemented with lactulose. The obtained results support the hypothesis that electro-activated whey has evident prebiotic effect compared to lactulose.

Independent and Combined Effects of Probiotics and Prebiotics as Supplements or Food-Rich Diets on a Propionic-Acid-Induced Rodent Model of Autism Spectrum Disorder.

The link between nutrition and autism spectrum disorder (ASD) as a neurodevelopmental condition, which is clinically presented as significant delays or deviations in interaction and communication, has provided a fresh point of view and signals that nutrition may play a role in the etiology of ASD, as well as playing an effective role in treatment by improving symptoms. In this study, 36 male albino rat pups were used. They were randomly divided into five groups. The control group was fed only a standard diet and water for the 30 days of the experiment. The second group, which served as a propionic acid (PPA)-induced rodent model of ASD, received orally administered PPA (250 mg/kg body weight (BW)) for 3 days, followed by feeding with a standard diet until the end of the experiment. The three other groups were given PPA (250 mg/kg body weight (BW)) for 3 days and then fed a standard diet and orally administered yogurt (3 mL/kg BW/day), artichokes (400 mL/kg BW/day), and a combination of Lacticaseibacillus rhamnosus GG at 0.2 mL daily (1 × 109 CFU; as the probiotic of yogurt) and luteolin (50 mg/kg BW/day; as the major antioxidant and anti-inflammatory ingredient of artichokes) for 27 days. Biochemical markers, including gamma-aminobutyric acid (GABA), reduced glutathione (GSH), glutathione peroxidase (GPx1), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-10 (IL-10), were measured in brain homogenates in all groups. The data showed that while PPA demonstrated oxidative stress and neuroinflammation in the treated rats, yogurt, Lacticaseibacillus rhamnosus GG as a probiotic, and luteolin as a prebiotic ingredient in artichokes were effective in alleviating the biochemical features of ASD. In conclusion, nutritional supplementation seems to be a promising intervention strategy for ASD. A combined dietary approach using pro- and prebiotics resulted in significant amelioration of most of the measured variables, suggesting that multiple interventions might be more relevant for the improvement of biochemical autistic features, as well as psychological traits. Prospective controlled trials are needed before recommendations can be made regarding the ideal ASD diet.