Coaggregation Ability Research Articles

Cellulolytic Bacillus Strain: Production Optimization Using Wheat Bran under Solid-State Fermentation and Investigation of Its Probiotic Potential

Attention to the association of cellulolytic bacteria with probiotic potential as an additive in animal feeding has risen in the last decades. Such additive use in livestock feed is essential in improving animal health, growth, and production performances. This study was designed to identify probiotic characteristics and test the cellulolytic ability of Bacillus strains isolated from the dromedary gastrointestinal tract. Thus, thirty-two Bacillus strains were tested for their cellulolytic ability on cellulose Congo-red agar media. Among the isolates, only the strain D1B3 showed the largest degradation zone (2.4 cm) and was identified as Bacillus amyloliquefacians by 16S rRNA gene sequence analysis. Solid-state fermentation (SSF) retained this strain for cellulase and biomass production using wheat bran as a substrate. The fermentation was optimized through a central composite design, by exploring three factors: incubation temperature, moisture ratio, and pH. Biomass and cellulose enzyme activity were selected as responses and corresponding regression coefficients were calculated. The optimal parameters were: liquid-to-solid ratio (1.19%), pH buffer (6.2), and incubation temperature (36.99 °C) to obtain the highest level of biomass and cellulose enzyme activity reaching a value of 9.828 log CFU/g and 0.0144 g/L.min, respectively. The potentiality of Bacillus amyloliquefacians D1B3 as a probiotic was examined in vitro. It also showed antimicrobial activity against Pseudomonas aeruginosa 9027, Klebsiella pneumoniae, and Escherichia coli 10536. The isolate tolerates low pH and bile salt (0.3% Oxygall). The hydrophobicity and coaggregation abilities were 1.7% and 69.79%, respectively. The results indicated that Bacillus amyloliquefacians D1B3 could be a potential probiotic additive for improving in vitro fermentation of wheat bran and suggests the possibility of combining the probiotic attributes of this strain with its cellulolytic ability to enhance the rumen fermentation of animal feed.

In-vitro evaluation of the probiotic potential and the fermentation profile of Pediococcus and Enterococcus strains isolated from Moroccan camel milk.

The promotion of human health through natural approaches like functional foods and probiotics is in high demand. The medicinal plants are the major feed of Moroccan dromedary, which improves the functional properties of their milk. A few studies have reported the probiotic and functional aptitudes of lactic acid bacteria (LAB) of this milk. In this context, our study aimed to identify LAB isolated from Moroccan raw camel milk and investigate their probiotic features and their fermentation profile. The molecular identification of twelve isolates indicated that they belong to Pediococcus pentosaceus, Enterococcus faecium, and Enterococcus durans. All LAB strains displayed high tolerance to gastrointestinal conditions (survival rate of 31.85-96.52% in pH 2.5, 35.23-99.05% in 0.3 bile salts, and 26.9-90.96% in pepsin), strong attachment abilities (auto-aggregation and hydrophobicity ranged from 28.75 to 95.9% and from 80.47 to 96.37%, respectively), and high co-aggregation ability with pathogenic bacteria. Importantly, they did not present antibiotic resistance or hemolytic activity. Our LAB strains demonstrated antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Bacillus subtilis, and Salmonella enterica. Moreover, they could acidify cow milk (ΔpH of 2.55 after 24h) and improve its antioxidant ability (inhibition of 36.77% of DPPH). Based on the multivariate analysis, Pediococcus pentosaceus Pd24, Pd29, Pd38, Enterococcus faecium Ef18, and Enterococcus durans Ed22 were selected as the most promising probiotics. Therefore, we propose that Pediococcus pentosaceus isolated from camel milk could be used as potential probiotic strains and/or starter cultures in functional milk fermentation.

Antimicrobial and immunoregulatory effects of Lactobacillus delbrueckii 45E against genitourinary pathogens

BackgroundLactobacilli are essential microbiota that maintain a healthy, balanced vaginal environment. Vaginitis is a common infection in women during their reproductive years. Many factors are associated with vaginitis; one of them is the imbalance of microbiota in the vaginal environment. This study aimed to evaluate the antimicrobial properties of Lactobacillus delbrueckii 45E (Ld45E) against several species of bacteria, namely, Group B Streptococcus (GBS), Escherichia coli, Klebsiella spp., and Candida parapsilosis, as well as to determine the concentration of interleukin-17 (IL-17) in the presence of Ld45E.MethodsThe probiotic characteristics of Ld45E were evaluated by examining its morphology, pH tolerance, adhesive ability onto HeLa cells, hemolytic activity, antibiotic susceptibility, and autoaggregation ability. Then, the antimicrobial activity of Ld45E was determined using Ld45E culture, cell-free supernatant, and crude bacteriocin solution. Co-aggregation and competition ability assays against various pathogens were conducted. The immunoregulatory effects of Ld45E were analyzed by measuring the proinflammatory cytokine IL-17. A p-value less than 0.05 was considered statistical significance.ResultsLd45E is 3–5 mm in diameter and round with a flat-shaped colony. pH 4 and 4.5 were the most favorable range for Ld45E growth within 12 h of incubation. Ld45E showed a strong adhesion ability onto HeLa cells (86%) and negative hemolytic activities. Ld45E was also sensitive to ceftriaxone, cefuroxime, ciprofloxacin, and doxycycline. We found that it had a good autoaggregation ability of 80%. Regarding antagonistic properties, Ld45E culture showed strong antimicrobial activity against GBS, E. coli, and Klebsiella spp. but only a moderate effect on C. parapsilosis. Cell-free supernatant of Ld45E exerted the most potent inhibitory effects at 40 °C against all genital pathogens, whereas bacteriocin showed a robust inhibition at 37 °C and 40 °C. The highest co-aggregation affinity was observed with GBS (81%) and E. coli (40%). Competition ability against the adhesion of GBS (80%), E. coli (76%), Klebsiella (72%), and C. parapsilosis (58%) was found. Ld45E was able to reduce the induction of the proinflammatory protein IL-17.ConclusionsLd45E possessed antimicrobial and immunoregulatory properties, with better cell-on-cell activity than supernatant activity. Thus, Ld45E is a potential probiotic candidate for adjunct therapy to address vaginal infections.

A novel microplate-based spectrophotometric method for the quantitative assessment of freshwater bacterial coaggregation kinetics

Coaggregation, the specific recognition and adhesion of genetically distinct bacteria, is proposed to contribute to the development of freshwater biofilms. This work aimed to develop a microplate-based system to measure and model the kinetics of freshwater bacterial coaggregation. Blastomonas natatoria 2.1 and Micrococcus luteus 2.13 were evaluated for coaggregation ability using 24-well microplates containing novel dome shaped wells (DSWs) and standard flat-bottom wells. Results were compared to a tube-based visual aggregation assay. The DSWs facilitated the reproducible detection of coaggregation via spectrophotometry and the estimation of coaggregation kinetics using a linked mathematical model. Quantitative analysis using DSWs was more sensitive than the visual tube aggregation assay and subject to substantially less variation than flat-bottom wells. Collectively these results demonstrate the utility of the DSW-based method and improve upon the current toolkit for studying freshwater bacterial coaggregation.

Characterization of adhesion, anti-adhesion, co-aggregation, and hydrophobicity of Helicobacter pylori and probiotic strains

ObjectivesTo characterize the adhesion ability of nine Helicobacter pylori strains and eight probiotics in human oral keratinocyte cells (H357 cells) in comparison to intestinal cells (Caco-2 and HIEC-6 cells). Subsequently, the anti-adhesion and co-aggregation abilities of the selected probiotic strains on H. pylori strains were investigated. MethodsNine H. pylori strains, including H. pylori ATCC43504 (type strain), and 8 clinical strains, were isolated from oral samples of three patients (one non-disease, one gastritis patient, and one gastric cancer patient). Eight selected probiotic strains were used, as follows: Lacticaseibacillus paracasei SD1, Lacticaseibacillus rhamnosus SD4, L. rhamnosus SD11, Limosilactobacillus fermentum SD7, L. rhamnosus GG, Limosilactobacillus reuteri ATCC-PTA6475, Lacticaseibacillus casei Shirota, and L. paracasei CNCM I-1572. The adhesion and anti-adhesion abilities of H. pylori and the probiotic strains were investigated in H357, Caco-2, and HIEC-6 cells. Co-aggregation at various pHs, hydrophobicity, and surface receptors of the cell lines for H. pylori strains were examined. ResultsAll probiotic and H. pylori strains adhered to H357 significantly better than Caco-2, and HIEC-6 cells. Three probiotic strains (SD7, SD4, SD11) showed significantly higher adhesion than others. Of the clinical H. pylori strains, isolates from a gastric cancer patient had the highest adhesion ability to all of the cell lines tested. Probiotic strains that exhibited high adhesion ability provided high anti-adhesion and co-aggregation against H. pylori strains. Acidic conditions encouraged the co-aggregation of probiotics to H. pylori strains. ConclusionThis study provides information relating to the adhesion abilities of clinical H. pylori and probiotic strains to the oral mucosa when compared to the intestinal mucosa. Certain probiotic strains may be useful for the successful eradication of H. pylori infection via anti-adhesion and co-aggregation.

Effect of different drying methods on the functional properties of probiotics encapsulated using prebiotic substances.

Probiotics and prebiotics together work synergistically as synbiotics and confer various health benefits. Many studies on synbiotic foods only focus on the survival of probiotics but fail to evaluate their functional properties. The impact on functional properties should be explored to better understand its therapeutic efficacy. In this work, probiotics (Lactiplantibacillus plantarum NCIM 2083) were encapsulated with prebiotics (fructooligosaccharide + whey protein + maltodextrin) using spray-drying (SD), freeze-drying (FD), spray-freeze-drying (SFD), and refractance window-drying (RWD) techniques. Aggregation, intestinal adhesion, antagonistic activity, and bile salt hydrolase (BSH) activity of probiotics were studied before and after the encapsulation process. The SFD probiotics showed better aggregation ability (79% at 24-h incubation), on par with free cells (FC) (81% at 24-h incubation). The co-aggregation ability of encapsulated probiotics has drastic variations with each pathogenic strain. The adhesion ability of probiotics in chicken intestinal mucus was assessed by the crystal violet method, indicating no significant variations between FC and SFD probiotics. Also, encapsulated probiotics exhibit antagonistic activity (zone of inhibition in mm) against gut pathogens E. coli (11.33 to 17.34), S. faecalis (8.83 to 15.32), L. monocytogenes (13.67 to 18), S. boydii (12.17 to 15.5), and S. typhi (2.17 to 6.86). Overall, these studies confirm the significance and impact of various drying techniques on the functionality of encapsulated probiotics in synbiotic powders. KEY POINTS: • Understanding the relevance of processing effects on the functionality of probiotics. • Spray-freeze-dried probiotics showed superior functional properties. • The encapsulation process had no significant impact on bile salt hydrolase activity.

Isolation and Identification of Limosilactobacillus reuteri PSC102 and Evaluation of Its Potential Probiotic, Antioxidant, and Antibacterial Properties.

We isolated and characterized Limosilactobacillus reuteri PSC102 and evaluated its probiotic, antioxidant, and antibacterial properties. We preliminarily isolated 154 candidates from pig feces and analyzed their Gram nature, morphology, and lactic acid production ability. Based on the results, we selected eight isolates and tested their ability to produce digestive enzymes. Finally, we identified one isolate using 16S rRNA gene sequencing, namely, L. reuteri PSC102. We tested its probiotic properties in vitro, including extracellular enzyme activities, low pH and bile salt tolerance, autoaggregation and coaggregation abilities, adhesion to Caco-2 cells, antibiotic susceptibility, and hemolytic and gelatinase activities. Antioxidant activity was determined using 1-diphenyl-2-picrylhydrazyl and 2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical scavenging and reducing power assays. The antibacterial activity of this strain and its culture supernatant against enterotoxigenic Escherichia coli were evaluated using a time-kill assay and disk diffusion method, respectively. L. reuteri PSC102 exhibited tolerance toward low pH and bile salt and did not produce harmful enzymes or possess hemolytic and gelatinase activities. Its intact cells and cell-free extract exhibited potential antioxidant activities, and significantly inhibited the growth of enterotoxigenic E. coli. Our results demonstrate that L. reuteri PSC102 is a potential probiotic candidate for developing functional feed.

Potential oral probiotic Lactobacillus pentosus MJM60383 inhibits Streptococcus mutans biofilm formation by inhibiting sucrose decomposition

ABSTRACT Streptococcus mutans is known as a contributor to dental caries. In this work, Lactobacillus pentosus MJM60383 was selected for its strong antagonistic activity against S. mutans and was characterized by good oral probiotic properties including lysozyme tolerance, adhesive ability to oral cells, good aggregation (auto-aggregation, co-aggregation) ability, hydrogen peroxide production and inhibition of biofilm formation of S. mutans. L. pentosus MJM60383 also exhibited safety as a probiotic characterized by no hemolytic activity, no D-lactate production, no biogenic amine production, and susceptibility to antibiotics. Furthermore, the biofilm formation of S. mutans was also significantly inhibited by the supernatant of L. pentosus MJM60383. An anti-biofilm mechanism study revealed that sucrose decomposition and the production of water-insoluble exopolysaccharides by S. mutans were inhibited by the treatment with L. pentosus MJM60383 supernatant. Real-time PCR analysis indicated that the supernatant of L. pentosus MJM60383 significantly inhibited the mRNA expression of S. mutans glycosyltransferases, which synthesize glucan to construct biofilm architecture and mediate bacterial adherence. Our study demonstrated L. pentosus MJM60383 as a potential oral probiotic and revealed its anti-biofilm mechanism.

Probiotic Properties of Chicken-Derived Highly Adherent Lactic Acid Bacteria and Inhibition of Enteropathogenic Bacteria in Caco-2 Cells.

Lactic acid bacteria (LAB) as probiotic candidates have various beneficial functions, such as regulating gut microbiota, inhibiting intestinal pathogens, and improving gut immunity. The colonization of the intestine is a prerequisite for probiotic function. Therefore, it is necessary to screen the highly adherent LAB. In this study, the cell surface properties, such as hydrophobicity, auto-aggregation, co-aggregation, and adhesion abilities of the six chicken-derived LAB to Caco-2 cells were investigated. All six strains showed different hydrophobicity (21.18-95.27%), auto-aggregation (13.61-30.17%), co-aggregation with Escherichia coli ATCC 25922 (10.23-36.23%), and Salmonella enterica subsp. enterica serovar Typhimurium ATCC 13311 (11.71-39.35%), and adhesion to Caco-2 cells (8.57-26.37%). Pediococcus pentosaceus 2-5 and Lactobacillus reuteri L-3 were identified as the strains with strong adhesion abilities (26.37% and 21.57%, respectively). Moreover, these strains could survive in a gastric acid environment at pH 2, 3, and 4 for 3 h and in a bile salt environment at 0.1%, 0.2%, and 0.3% (w/v) concentration for 6 h. Furthermore, the cell-free supernatant of P. pentosaceus 2-5 and L. reuteri L-3 inhibited the growth of enteropathogenic bacteria and the strains inhibited the adhesion of these pathogens to Caco-2 cells. In this study, these results suggested that P. pentosaceus 2-5 and L. reuteri L-3, isolated from chicken intestines might be good probiotic candidates to be used as feed additives or delivery vehicles of biologically active substances.

Determination of safety status and probiotic properties ofEnterococcusstrains isolated from traditional cheeses in Turkey

This study aimed to evaluate the probiotic properties of Enterococcus strains isolated from Turkish traditional cheeses. Fifty-two Enterococcus spp. were taxonomically determined as follows: Enterococcus faecium (26), Enterococcus faecalis (18), Enterococcus durans (6), and Enterococcus italicus (2). The ability of isolates/strains to survive the harsh conditions (acidity and in-vitro gastric solution) of the gastrointestinal tract was established. They also showed auto-aggregation, hydrophobicity, and co-aggregation ability. Hydrophobicities of the strains were found between 0.8%-21%, 0.7%-56%, and 2%-63% for xylene, chloroform, and ethyl acetate, respectively. Autoaggregation values of the Enterococcus strains were 4%-20%, 7%-30%, and 36%-98% after 2, 4, and 24-h incubation, respectively. In this study, the Enterococcus strains tested showed co-aggregation ability with the Escherichia coli ATCC 25922, Salmonella Typhimurium ATCC 14028, and Staphylococcus aureus ATCC 25923. The results of PCR amplification revealed that only five strains possess virulence factor genes (gelE,asa1,cyl A,esp). We determined antibiotic resistance, biofilm forming abilities, and hemolytic activity for safety evaluation of strains. In this large and comprehensive study, we found that only few of Enterococcus strains have promising probiotic potential, among which E. faecalis ES1 and E. faecium EM1 showed the best probiotic properties (are the most promising probiotic candidates).

Probiotic of Lactiplantibacillus plantarum NWAFU-BIO-BS29 Isolated from Chinese Traditional Fermented Milk and Its Potential Therapeutic Applications Based on Gut Microbiota Regulation.

Lactic acid bacteria are one of the bioresources that can promote the host's health and have potential therapeutic applications. This study aimed to evaluate the probiotic properties of novel Lactiplantibacillus plantarum NWAFU-BISO-BS29 isolated in vitro from traditional Chinese fermented milk, assess its safety, and study its interaction with the gut microbiota using a BALB/c mouse model. The findings reveal that this strain had a high tolerance to gastric acidity (64.4%) and bile salts (19.83-87.92%) with remarkable auto-aggregation and co-aggregation abilities (33.01-83.96%), respectively. Furthermore, it lowered the cholesterol levels in dead cells (44.02%) and live cells (34.95%) and produced short-chain fatty acids (SCFAs). Likewise, it showed good antioxidant properties and strong antipathogen activity against Escherichia coli and Staphylococcus aureus with inhibition zones at 21 and 25 mm, respectively. The safety assessment results indicate that all of the virulence factor genes were not detected in the whole DNA; additionally, no hemolysis or resistance to antibiotics commonly used in food and feed was observed. Interestingly, the 16S rRNA gene sequencing of the mouse gut microbiota showed a marked alteration in the microbial composition of the administrated group, with a noticeable increase in Firmicutes, Patescibacteria, Campylobacterota, Deferribacterota, Proteobacteria, and Cyanobacteria at the phylum level. The modulation of gut microbial diversity significantly improved the production of SCFCs due to the abundance of lactobacillus genera, which was consistent with the functional gene predictive analysis and is believed to have health-promoting properties. Based on these results, our novel strain is considered a safe and good probiotic and could hold high potential to be used as a starter culture or to safely supplement functional foods as a probiotic and may provide new insights into therapeutic interventions.

Characterization of Leuconostoc mesenteroides MJM60376 as an oral probiotic and its antibiofilm activity.

Lactic acid bacteria have been widely used as probiotics for improving gut health. However, studies on oral probiotics were very limited. In this study, 67 lactic acid bacteria (LAB) were isolated from fermented food and screened for antagonistic activity against Streptococcus mutans, the causative pathogen of dental caries. Leuconostoc mesenteroides MJM60376 showed the highest antagonistic activity against S. mutans KCTC3065. L. mesenteroides MJM60376 also showed oral probiotic characteristics including weak acid production, lysozyme tolerance, adhesion to oral epithelial cell (YD-38), antibiotic susceptibility, and good coaggregation ability with S. mutans. Furthermore, the biofilm formation of S. mutans was significantly reduced when cocultured with L. mesenteroides. Scanning electron microscopy analysis showed that amounts of attached bacteria of S. mutans and network-like structures were significantly reduced by L. mesenteroides MJM60376. Cell-free supernatant (CFS) of L. mesenteroides MJM60376 also greatly inhibited biofilm formation of S. mutans from the adherent stage, the activity remained even after it was treated with catalase, trypsin, or pH neutralized. Expression levels of biofilm formation-related genes were significantly reduced in S. mutans when it was treated with the CFS of L. mesenteroides MJM60376. Therefore, L. mesenteroides MJM60376 has great potential to be used as a multifunctional ingredient.

The Synbiotic Effect of Probiotics and Dried Spirulina platensis or Phycocyanin on Biofilm Formation by Salmonella Typhimurium and Staphylococcus aureus.

This study aimed to evaluate the synbiotic effect of probiotics and dried Spirulina platensis or phycocyanin on autoaggregation, coaggregation, and the inhibition of biofilm formation by Salmonella Typhimurium and Staphylococcus aureus on 96-well microtiter plates and Human colon carcinoma cells-116 surfaces. The results showed that the probiotics strains cultured in the presence of S. platensis exhibited the highest autoaggregation values, ranging between 68.5 and 74.2% after 24 h. All probiotic strains with or without S. platensis and phycocyanin showed coaggregation abilities with S. Typhimurium and S. aureus. Interestingly, significant effect of S. platensis and phycocyanin supplementation was observed on the inhibition of the biofilm formation by the selected pathogens during the competition, exclusion, and displacement on abiotic and biotic surfaces.

Isolation and characterization of probiotic lactic acid bacteria from human breast milk

The lactic acid bacteria (LAB) isolated from human breast milk is known as probiotics and comprises numerous health benefits. This study aims to select and determine the species name of LAB based on the 16S rRNA gene, which has the potential to be the best indigenous probiotic. The method used included analysis of LAB resistance at acidic pH 2.0 and bile salts (0.5 %), antimicrobial activity against pathogenic microorganisms, and determining the autoaggregation properties. LAB isolates with the best ability in the analysis were then identified using a partial sequence of the 16S rRNA gene. The isolation and purification revealed eight LAB isolates with different parameters named as L19A, L19B, L19C, L19D, L19E, L19F, L19G, and L19H. Isolates L19A, L19E, and L19H have good tolerance ability against acid pH and bile salts, compared to others. Meanwhile, the L19H isolate had the strongest antimicrobial activity against pathogenic microorganisms E. coli ATCC 25922, S. aureus ATCC 25923, and C. albicans ATCC 11778, while the L19A had the highest hydrophobicity, autoaggregation, and coaggregation ability. Based on the partial sequence analysis of the 16S rRNA gene, the L19A, L19E, and L19H have similar values with L. casei, L. rhamnosus, and L. paracasei, respectively. These isolates belong to the L. casei group (LCG) from human breast milk, which can be used as an indigenic probiotic.

Physiological properties, survivability and genomic characteristics of Pediococcus pentosaceus for application as a starter culture

There is an increasing demand for the use of Pediococcus pentosaceus in the dairy industry due to its remarkable abilities as a probiotic or starter culture bacterium. This study determines the physiological, survivability and genomic properties of P. pentosaceus to evaluate its potential as a starter culture. The strain showed great tolerance in low pH and high concentrations of bile salts. A good coaggregation ability against pathogens ranging between 71% and 84%, as well as strong biofilm formation ability and remarkable resistance to antibiotics. Whole‐genome sequence results reveal the presence of various genes responsible for stress tolerance, adhesion, biofilm formation, acids, and bile salts tolerance. The findings of this study can be used as essential criteria for selecting P. pentosaceus as a starter culture.

Adhesion and aggregation properties of Lactobacillaceae strains as protection ways against enteropathogenic bacteria

The adhesion and aggregation are characteristic attributes of probiotic strains belonging to Lactobacillaceae genus. Due to these properties the host organisms can avoid colonisation of the intestinal tract by enteropathogenic bacteria. The presented research includes a comparison of the properties of various strains belonging to different Lactobacillaceae species and isolated from different sources The aim of this study was to investigate the ability of Lactocaseibacillus rhamnosus, Lactiplantibacillus plantarum, and Lactobacillus strains (L. acidophilus, L. gasseri, L. ultunensis) from probiotic products and clinical specimens to direct and competitive adherence to Caco-2 and HT-29 cell lines. Furthermore, the ability of lactobacilli and enteropathogenic bacteria, E. coli, E. faecalis, and S. Typhimurium, to auto- and co-aggregation was also investigated.The results showed that all tested strains adhered to Caco-2 and HT-29 cell lines. Though, the factor of adhesion depended on the species and origin of the strain. L. rhamnosus strains showed a lowest degree of adherence as compared to L. plantarum and Lactobacillus sp. strains. On the other side both, L. rhamnosus and L. acidophilus strains reduced the pathogenic bacteria in competition adherence test most effectively. All tested lactobacilli strains were characterised by auto- and co-aggregation abilities, to various degrees. The properties of Lactobacillaceae strains analysed in this study, like adhesion abilities, competitive adherence, auto- and co-aggregation, may affect the prevention of colonisation and elimination of pathogenic bacteria in gastrointestinal tract.

Screening of potential probiotic lactic acid bacteria with antimicrobial properties and selection of superior bacteria for application as biocontrol using machine learning models

144 lactic acid bacteria (LAB) strains were screened to find those with the best antimicrobial activity and potential probiotic properties. Firstly, bacteria were tested for resistance to low pH and high bile salt, antimicrobial properties against the common agent of bacterial contamination in plant tissue cultures. Then, the selected strains were assayed for probiotic characteristics including autoaggregation, coaggregation, hydrophobicity, biofilm formation, and adhesion ability. Finally, ranking and selection of probiotic potential bacteria for harnessing as antibacterial agents in plant tissue cultures were performed using supervised machine learning models. The highest values of autoaggregation percentages were observed for Lactobacillus paracasei S23 (96.1%), while the highest coaggregation values with Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes were observed for strains Lactobacillus plantarum S57 (63.4%), L. plantarum S70 (57.4%), and Lactobacillus casei S81 (58.6%), respectively. Principal component analysis (PCA) and machine learning algorithms allowed to select and prioritizing of 15 out of 144 LAB strains, based on their probiotic potential properties or their antimicrobial activity, of which four strains including Lb. paracasei S23, Lb. plantarum S70, Lb. casei S81, and Lb. plantarum S57 were proposed as appropriate antimicrobial candidates for harnessing in plant tissue cultures.

E-Cigarette Aerosol Exposure Favors the Growth and Colonization of Oral Streptococcus mutans Compared to Commensal Streptococci.

ABSTRACTE-cigarettes (e-cigs) have drastically increased in popularity during the last decade, especially among teenagers. While recent studies have started to explore the effect of e-cigs in the oral cavity, little is known about their effects on the oral microbiota and how they could affect oral health and potentially lead to disease, including periodontitis and head and neck cancers. To explore the impact of e-cigs on oral bacteria, we selected members of the genus Streptococcus, which are abundant in the oral cavity. We exposed the commensals Streptococcus sanguinis and Streptococcus gordonii and the opportunistic pathogen Streptococcus mutans, best known for causing dental caries, to e-liquids and e-cig aerosols with and without nicotine and with and without menthol flavoring and measured changes in growth patterns and biofilm formation. Our results demonstrate that e-cig aerosols hindered the growth of S. sanguinis and S. gordonii, while they did not affect the growth of S. mutans. We also show that e-cig aerosols significantly increased biofilm formation by S. mutans but did not affect the biofilm formation of the two commensals. We found that S. mutans exhibits higher hydrophobicity and coaggregation abilities along with higher attachment to OKF6 cells than S. sanguinis and S. gordonii. Therefore, our data suggest that e-cig aerosols have the potential to dysregulate oral bacterial homeostasis by suppressing the growth of commensals while enhancing the biofilm formation of the opportunistic pathogen S. mutans. This study highlights the importance of understanding the consequences of e-cig aerosol exposure on selected commensals and pathogenic species. Future studies modeling more complex communities will provide more insight into how e-cig aerosols and vaping affect the oral microbiota.IMPORTANCE Our study shows that e-cigarette aerosol exposure of selected bacteria known to be residents of the oral cavity hinders the growth of two streptococcal commensals while enhancing biofilm formation, hydrophobicity, and attachment for the pathogen S. mutans. These results indicate that e-cigarette vaping could open a niche for opportunistic bacteria such as S. mutans to colonize the oral cavity and affect oral health.

The In Vitro Analysis of Postbiotics in Functional Labneh to Be Used as Powerful Tool to Improve Cell Surfaces Properties and Adherence Potential of Probiotic Strains

Postbiotics are functional bioactive substances manufactured during fermentation in a food matrix, which can be used to improve human health, but their influence on the adhesion potential and physicochemical cell surface of probiotics is still unclear. We examined the postbiotic influence produced by Escherichia coli Nissle 1917 in functional labneh on cell surface properties (auto-aggregation, hydrophobicity, and co-aggregation) and the adhesion capacities of three probiotic strains. The most commonly detected effects of probiotics, particularly Lsyn−7, were an increase in auto-aggregation, hydrophobicity, co-aggregation, and adhesion ability of the tested strains. Lactobacillus rhamnosus with Lsyn−7 (59%) presented the highest hydrophobicity, whereas the least adhesion to xylene was detected in L. rhamnosus with LHM. Lactobacillus casei with Lsyn−7 showed the highest auto-aggregation after 24 h (60.55%). Moreover, it also has a strong adhesion to Caco-2 cells and effectively prevents the binding of Salmonella Typhimurium to Caco-2 cells. Lactobacillus plantarum with Lsyn−7 presented the strongest co-aggregation with Staphylococcus aureus (85.1%), S. typhimurium (85. 02%) and Listeria monocytogenes (77.4%). The adherence potential of tested probiotic strains was highly correlated with auto-aggregation, hydrophobicity, co-aggregation, and competitive inhibition of L. monocytogenes and S. typhimurium. The findings suggest that Lsyn−7 can be a candidate to promote the adhesion potential of selected probiotic strains. For the reason that the application of probiotic strains has been more interested in their positive influences in the gastrointestinal tract, it is essential to use some functional compounds, such as postbiotics, to improve adhesion abilities and cell surface properties in terms of bacterial binding.

Comparative Analysis of Fermentation Conditions on the Increase of Biomass and Morphology of Milk Kefir Grains

Kefir grains represent a symbiotic association group of yeasts, lactic acid bacteria and acetic acid bacteria within an exopolysaccharide and protein matrix known as kefiran. The mechanism of growth of a biomass of kefir after successive fermentations and optimal conditions is not well understood yet. Biomass growth kinetics were determined to evaluate the effects of temperatures (10 °C to 40 °C) and different substrates, such as monosaccharides (fructose, galactose, glucose), disaccharides (lactose, saccharose) and polysaccharides (Agave angustifolia fructans) at 2%, in reconstituted nonfat milk powder at 10% (w/v) and inoculated with 2% of milk kefir grain (105 CFU/g), after determining the pH kinetics. The best conditions of temperature and substrates were 20 °C and fructans and galactose. An increase in cells, grain sizes and a change in the morphology of the granules with the best substrates were observed using environmental scanning electron microscopy, confocal laser scanning microscopy and Image Digital Analysis (IDA). Kefir grains with agave fructans as their carbon source showed the higher fractal dimension (2.380), related to a greater co-aggregation ability of LAB and yeasts, and increase the formation of exopolysaccharides and the size of the kefir grains, which opens new application possibilities for the use of branched fructans as a substrate for the fermentation of milk kefir grains for the enhancement of cellular biomasses and exopolysaccharide production, as well as IDA as a characterization tool.