Cell Surface Characteristics Research Articles

Directed evolution of the multicopper oxidase laccase for cell surface proximity labeling and electron microscopy.

Enzymes that oxidize aromatic substrates have shown utility in a range of cell-based technologies including live cell proximity labeling (PL) and electron microscopy (EM), but are associated with drawbacks such as the need for toxic H2O2. Here, we explore laccases as a novel enzyme class for PL and EM in mammalian cells. LaccID, generated via 11 rounds of directed evolution from an ancestral fungal laccase, catalyzes the one-electron oxidation of diverse aromatic substrates using O2 instead of toxic H2O2, and exhibits activity selective to the surface plasma membrane of both living and fixed cells. We show that LaccID can be used with mass spectrometry-based proteomics to map the changing surface composition of T cells that engage with tumor cells via antigen-specific T cell receptors. In addition, we use LaccID as a genetically-encodable tag for EM visualization of cell surface features in mammalian cell culture and in the fly brain. Our study paves the way for future cell-based applications of LaccID.

An Acinetobacter baumannii nasal carriage isolate recovered from an asymptomatic patient in Vietnam is extensively antibiotic resistant and produces a rare K71 type capsule.

The majority of Acinetobacter baumannii genomes sequenced and analyzed to develop an understanding of extensively drug-resistant (XDR) isolates belong to the globally disseminated CC2 clonal complex. While XDR isolates belonging to rarer lineages are often unexplored, detailed analyses could provide novel insights into the spread of resistance, as well as cell surface features such as the CPS that determine the specificity of non-antibiotic therapeutics required to treat XDR infections that resist antimicrobial chemotherapy. Here, we describe the properties of an XDR asymptomatic nasal carriage isolate recovered in Vietnam that belongs to ST142, a rarely encountered sequence type. We report the resistance profile and correlate this with detected resistance determinants. We also solve the structure of the CPS and reveal its relationship with CPS produced by other A. baumannii isolates.

Bacterial cell surface characterization by phage display coupled to high-throughput sequencing

The remarkable capacity of bacteria to adapt in response to selective pressures drives antimicrobial resistance. Pseudomonas aeruginosa illustrates this point, establishing chronic infections during which it evolves to survive antimicrobials and evade host defenses. Many adaptive changes occur on the P. aeruginosa cell surface but methods to identify these are limited. Here we combine phage display with high-throughput DNA sequencing to create a high throughput, multiplexed technology for surveying bacterial cell surfaces, Phage-seq. By applying phage display panning to hundreds of bacterial genotypes and analyzing the dynamics of the phage display selection process, we capture important biological information about cell surfaces. This approach also yields camelid single-domain antibodies that recognize key P. aeruginosa virulence factors on live cells. These antibodies have numerous potential applications in diagnostics and therapeutics. We propose that Phage-seq establishes a powerful paradigm for studying the bacterial cell surface by identifying and profiling many surface features in parallel.

Functional insights into the interaction of Lactobacillus spp. and potentially protective agents

The research addresses the critical need to explore how potentially protective agents (PPAs) influence the cellular characteristics of probiotic bacteria, with a focus on strain-specific responses. This study aimed to reveal the effect of 48-h exposure of the Lactobacillus spp. strains to selected PPAs on the viability, metabolic activity, stress response, surface properties, and physicochemical stability of bacterial suspensions. Results provide new insights into the potential risks or benefits of using the tested PPAs in probiotic production based on the analyzed strains, offering valuable information on their impacts on bacterial cells. Trehalose (TRE) and vitamin C (VC) notably impact the metabolic activity and surface characteristics of cells, with TRE improving viability and VC changing surface roughness and particle size distribution (PSD), suggesting a delicate balance between stress induction and viability enhancement. Inulin (IN) decreased metabolic changes, pointing the complex interplay between PPAs and bacterial metabolism. Results of zeta potential and PSD measurements revealed that other PPAs can either promote uniformity or induce aggregation of the bacterial suspensions. Findings have shown significant strain-specific effects of PPAs on bacterial viability and functionality, suggesting that their careful selection can improve bacterial health. Future directions include exploring the molecular mechanisms behind PPAs' effects and their practical applicability in real food systems.

Probiotic potential of acetic acid bacteria isolated from kombucha in New Zealand in vitro

Kombucha is a fermented tea beverage containing live microorganisms, mostly beneficial strains of yeast and acetic acid bacteria (AAB), but empirical evidence is limited supporting the probiotic potential of kombucha. This study reports the in vitro probiotic potential of 36 AAB strains isolated from three Kombucha samples commercially available in New Zealand. Nine representative AAB strains, belonging to three species (Komagataeibacter rhaeticus, Acetobacter musti and Gluconobacter potus), were examined for their primary probiotic characteristics such as tolerance to bile salts, NaCl, and low pH, and temperature. Three non-cellulose forming strains (A. musti LOAAB1, G. potus LOAAB2 and G. potus GBAAB3) were assayed for their cell surface characteristics such as auto-aggregation, co-aggregation with pathogenic bacteria, and hydrophobicity. Antimicrobial, antioxidant activities and enzymatic activities were also investigated for the strains of interest. Results indicated that nine strains were able to grow under low pH in the presence of bile salts, suggesting their potential to survive in the human gut. Six K. rhaeticus strains produced cellulosic pellicles, a potential source of prebiotics for beneficial bacteria. The three AAB strains LOAAB1, LOAAB2 and GBAAB3 showed promising cell surface characteristics, such as auto-aggregation rates (>80 %), co-aggregation with four pathogenic bacteria (13.24–43.47 %), hydrophobicity (42.12 % to 50.20 %), and antioxidant activities (>90 %). All nine strains tested negative for enzymatic activities (haemolytic, proteolytic, phospholipase, and gelatinase), suggesting that they are safe to consume. Together, these data indicate the potential for the three AAB strains to be further investigated as probiotic sources with more in vivo tests for applications in the food and beverages industry.

Inhibitory effect of Lonicera japonica flos on Streptococcus mutans biofilm and mechanism exploration through metabolomic and transcriptomic analyses.

Streptococcus mutans was the primary pathogenic organism responsible for dental caries. Lonicera japonica flos (LJF) is a traditional herb in Asia and Europe and consumed as a tea beverage for thousands of years. The inhibitory effect and mechanism of LJF on biofilm formation by S. mutans was investigated. The active extracts of LJF were validated for their inhibitory activity by examining changes in surface properties such as adherence, hydrophobicity, auto-aggregation abilities, and exopolysaccharides (EPS) production, including water-soluble glucan and water-insoluble glucan. LJF primarily inhibited biofilm formation through the reduction of EPS production, resulting in alterations in cell surface characteristics and growth retardation in biofilm formation cycles. Integrated transcriptomic and untargeted metabolomics analyses revealed that EPS production was modulated through two-component systems (TCS), quorum sensing (QS), and phosphotransferase system (PTS) pathways under LJF stress conditions. The sensing histidine kinase VicK was identified as an important target protein, as LJF caused its dysregulated expression and blocked the sensing of autoinducer II (AI-2). This led to the inhibition of response regulator transcriptional factors, down-regulated glycosyltransferase (Gtf) activity, and decreased production of water-insoluble glucans (WIG) and water-soluble glucans (WSG). This is the first exploration of the inhibitory effect and mechanism of LJF on S. mutans, providing a theoretical basis for the application of LJF in functional food, oral health care, and related areas.

Antibiofilm activity of d-limonene against spoilage Bacillus amyloliquefaciens

This study aimed to determine the efficacy of d-limonene in preventing biofilm formation by Bacillus amyloliquefaciens, a soy food spoilage bacterium. The minimal biofilm inhibitory concentrations (MBICs) of d-limonene against two strains of B. amyloliquefaciens were determined using a twofold dilution method. Furthermore, the ability of d-limonene to eradicate preformed biofilms was evaluated using the TTC reduction method. Subsequent analyses investigated the effects of d-limonene on biofilm morphology, cell motility, cell surface characteristics, and the composition of the biofilm matrix. The MBICs of d-limonene for B. amyloliquefaciens strains DY1a and DY1b were determined to be 24 and 36 μL/mL, respectively. At 2 MBIC, d-limonene eradicated 63.67% of the DY1a biofilms and 52.63% of the DY1b biofilms. Significantly, d-limonene markedly reduced swimming and swarming motility at concentrations ranging from 1/2 to 1 MBIC. In particular, at 1/2 MBIC, d-limonene decreased swimming and swarming abilities by 85.88% and 37.84%, respectively, for strain DY1a and by 75.00% and 95.07%, respectively, for strain DY1b. Furthermore, d-limonene suppressed the biosynthesis of polysaccharides and proteins in the extracellular biofilm matrix and reduced cell surface hydrophobicity, surface charge, and autoaggregation. These findings suggest that d-limonene could be an effective antibiofilm agent for the control of B. amyloliquefaciens in the soybean food industry.

The Probiotic Properties and Safety of Limosilactobacillus mucosae NK41 and Bifidobacterium longum NK46.

Probiotics should possess specific properties to exert beneficial effects, and their safety must be ensured for human consumption. The purpose of this study was to evaluate the probiotic properties and safety of Limosilactobacillus mucosae NK41 and Bifidobacterium longum NK46 isolated from human feces in vitro. Both strains exhibited high resistance to simulated gastrointestinal fluid. Furthermore, probiotic-related cell surface characteristics including auto-aggregation and cell surface hydrophobicity were assessed by measuring the absorbance at a wavelength of 600 nm, which demonstrated good auto-aggregation ability and affinity for xylene, indicating their effective adhesion to Caco-2 cells. In addition, hemolytic, gelatinase, and β-glucuronidase activities were found to be negative in both strains. The susceptibility to nine commonly used antibiotics was assessed using the broth macrodilution method, which demonstrated that both strains were susceptible to all tested antibiotics. Furthermore, L. mucosae NK41 and B. longum NK46 produced significantly higher levels of L-lactate (71.8 ± 0.7% and 97.8 ± 0.4%) than D-lactate (28.2 ± 0.7% and 2.2 ± 0.4%, respectively). Using PCR amplification to investigate genes associated with virulence factors, we found that neither strain harbored any virulence genes. These findings suggest that L. mucosae NK41 and B. longum NK46 have the potential to be used as probiotics and are considered safe for human consumption.

Aggregation and adhesion ability of various probiotic strains and Candida species: An in vitro study

Background/purposeThe ability of probiotics to inhibit Candida adhesion is a crucial characteristic that prevents Candida colonization and infection progression. This study aimed to explore aggregation, adhesion, and cell surface characterization of probiotic and Candida strains and to evaluate the effect of probiotics and their cell-free supernatants (CFSs) as postbiotics on Candida adhesion to human oral keratinocytes. Materials and methodsEight probiotic strains and five reference Candida strains were tested for autoaggregation, coaggregation, adhesin on human oral keratinocytes (H357), and cell surface properties. The anti-Candida adhesion activities of probiotic strains and CFSs were investigated. ResultsThe results showed that most probiotics exhibited high adhesion to H357 cells, specifically oral probiotic Lacticaseibacillus rhamnosus SD4, Limosilactobacillus fermentum SD7, and L. rhamnosus SD11, and adhesion ability of probiotic strains was strongly related to their autoaggregation, cell surface charges, and hydrophobicity. Candida strains also revealed a high level of adhesion to H357 cells. Candida albicans and Candida glabrata showed significantly higher adhesion abilities than others. After a combination of Candida with probiotics or their CFSs, Candida adhesion was significantly reduced. The anti-Candida adhesion property of probiotics was strongly related to their autoaggregation, coaggregation, and adhesion abilities. ConclusionThis study demonstrated that oral probiotic strains may be useful probiotics for preventing and treating oral candidiasis due to their high ability of aggregation, adhesion, and anti-Candida adhesion to H357 cells.

Evaluation of the probiotic potential of yeast isolated from kombucha in New Zealand

The current study investigated the in vitro probiotic potential of yeast isolated from kombucha, a tea beverage fermented with a symbiotic culture of acetic acid bacteria and yeast. A total of 62 yeast strains were previously isolated from four different commercial kombucha samples sold in New Zealand. Fifteen representative isolates belonging to eight different species were evaluated for their growth under different conditions (temperature, low pH, concentrations of bile salts, and NaCl). Cell surface characteristics, functional and enzymatic activities of the selected strains were also studied in triplicate experiments. Results showed that six strains (Dekkera bruxellensis LBY1, Sachizosaccharomyces pombe LBY5, Hanseniaspora valbyensis DOY1, Brettanomyces anomalus DOY8, Pichia kudraivzevii GBY1, and Saccharomyces cerevisiae GBY2) were able to grow under low-acid conditions (at pH 2 and pH 3) and in the presence of bile salts. This suggests their potential to survive passage through the human gut. All 15 strains exhibited negative enzymatic activity reactions (haemolytic, gelatinase, phospholipase, and protease activities), and thus, they can be considered safe to consume. Notably, two of the fifteen strains (Pichia kudraivzevii GBY1 and Saccharomyces cerevisiae GBY2) exhibited desirable cell surface hydrophobicity (64.60–83.87%), auto-aggregation (>98%), co-aggregation, resistance to eight tested antibiotics (ampicillin, chloramphenicol, colistin sulphate, kanamycin, nalidixic acid, nitrofurantoin, streptomycin, and tetracycline), and high levels of antioxidant activities (>90%). Together, our data reveal the probiotic activities of two yeast strains GBY1 and GBY2 and their potential application in functional food production.

THE EFFECT OF CHRONOLOGICAL AGE AND GENDER AND DIFFERENT SOURCE OF TISSUE ON THE IN VITRO PROPERTIES OF MESENCHYMAL STROMAL CELLS

AbstractObjectivesOsteoarthritis is a common articular cartilage disorder and causes a significant global disease burden. Articular cartilage has a limited capacity of repair and there is increasing interest in the use of cell-based therapies to facilitate repair including the use of Mesenchymal Stromal Cells (MSCs). There is some evidence in the literature that suggests that advancing age and gender is associated with declining MSC function, including reduced proliferation and differentiation potential, and greater cellular apoptosis. In our study, we first performed a systematic review of the literature to determine the effects of chronological age and gender on the in vitro properties of MSCs, and then performed a laboratory study to investigate these properties.Methods and ResultsWe initially conducted a PRISMA systematic review of the literature to review the evidence base for the effects of chronological age and gender on the in vitro properties of MSCs including cell numbers, expansion, cell surface characterization and differentiation potential. This was followed by laboratory-based experiments to assess these properties. Compare the extent of the effect of age on MSC cell marker expression, proliferation and pathways. Tissue from patients undergoing total knee replacement surgery was used to isolate MSCs from the synovium, fat pad and bone fragments using a method developed in our laboratory. The growth kinetics was determined by calculating the population doublings per day. Following expansion in culture, MSCs at P2 were characterised for a panel of cell surface markers using flow cytometry. The cells were positive for CD73, CD90 and CD105, and negative for antibody cocktail (eg included CD34, CD45). The differentiation potential of the MSCs was assessed through tri-lineage differentiation assays. At P2 after extracting RNA, we investigate the gene analysis using Bulk seq. Clear differences between the younger and older patients and gender were indicated.ConclusionsChronological age and gender-related changes in MSC function have important implications on the use of these cells in clinical applications for an ageing population. The results from this study will be used to plan further work looking at the effects of chronological age and gender on cellular senescence and identify pathways that could be targeted to potentially reverse any age and gender-related changes.Declaration of Interest(b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project.

The Effects of Chronological Age on the Chondrogenic Potential of Mesenchymal Stromal Cells: A Systematic Review.

Tissue engineering and cell therapy for regenerative medicine have great potential to treat chronic disorders. In musculoskeletal disorders, mesenchymal stromal cells (MSCs) have been identified as a relevant cell type in cell and regenerative strategies due to their multi-lineage potential, although this is likely to be a result of their trophic and immunomodulatory effects on other cells. This PRISMA systematic review aims to assess whether the age of the patient influences the chondrogenic potential of MSCs in regenerative therapy. We identified a total of 3027 studies after performing a search of four databases, including Cochrane, Web of Science, Medline, and PubMed. After applying inclusion and exclusion criteria, a total of 14 papers were identified that were reviewed, assessed, and reported. Cell surface characterization and proliferation, as well as the osteogenic, adipogenic, and chondrogenic differentiation, were investigated as part of the analysis of these studies. Most included studies suggest a clear link between aged donor MSCs and diminished clonogenic and proliferative potential. Our study reveals a heterogeneous and conflicting range of outcomes concerning the chondrogenic, osteogenic, and adipogenic potential of MSCs in relation to age. Further investigations on the in vitro effects of chronological age on the chondrogenic potential of MSCs should follow the outcomes of this systematic review, shedding more light on this complex relationship.

Probiotic and functional potential of lactic acid bacteria isolated from pulque and evaluation of their safety for food applications.

Pulque is a traditional Mexican non-distilled alcoholic beverage to which several beneficial functions are attributed, mainly associated with gastrointestinal health, which can be explained by the presence of probiotic bacteria in its microbiota. Therefore, the objective of this work was to evaluate the safety, probiotic activity, and functional characteristics of seven strains of lactic acid bacteria (LAB) isolated from pulque using the probiotic strain Lactobacillus acidophilus NCFM as control. The LAB isolates were identified by 16S rRNA sequencing and MALDI Biotyper® MS as belonging to three different Lactobacillaceae genera and species: Lactiplantibacillus plantarum, Levilactobacillus brevis and Lacticaseibacillus paracasei. Most strains showed resistance to gastric juice, intestinal juice and lysozyme (10 mg/L). In addition, all strains exhibited bile salt hydrolase (BSH) activity and antibacterial activity against the pathogenic strain Listeria monocytogenes. Additionally, cell surface characteristics of LAB were evaluated, with most strains showing good hydrophobicity, auto-aggregation, and co-aggregation towards enteropathogenic Escherichia coli and L. monocytogenes. In terms of safety, most of the strains were sensitive to the tested antibiotics and only the Lact. paracasei UTMB4 strain amplified a gene related to antibiotic resistance (mecA). The strains Lact. plantarum RVG2 and Lact. plantarum UTMB1 presented γ-hemolytic activity, and the presence of the virulence-related gene agg was identified only in UTMB1 strain. Regarding functional characterization, the tested bacteria showed good β-galactosidase activity, antioxidant activity and cholesterol reduction Based on principal component analysis (PCA) and heat mapping, and considering the strain Lact. acidophilus NCFM as the probiotic reference, the strains Lacticaseibacillus paracasei UTMB4, Lactiplantibacillus plantarum RVG4 and Levilactobacillus brevis UTMB2 were selected as the most promising probiotic strains. The results of this study highlighted the probiotic, functional and safety traits of LAB strains isolated from pulque thus supporting the health benefits attributed to this ancestral beverage.

Recent Advances in Microbial-Assisted Remediation of Cadmium-Contaminated Soil.

Soil contamination with cadmium (Cd) is a severe concern for the developing world due to its non-biodegradability and significant potential to damage the ecosystem and associated services. Industries such as mining, manufacturing, building, etc., rapidly produce a substantial amount of Cd, posing environmental risks. Cd toxicity in crop plants decreases nutrient and water uptake and translocation, increases oxidative damage, interferes with plant metabolism and inhibits plant morphology and physiology. However, various conventional physicochemical approaches are available to remove Cd from the soil, including chemical reduction, immobilization, stabilization and electro-remediation. Nevertheless, these processes are costly and unfriendly to the environment because they require much energy, skilled labor and hazardous chemicals. In contrasting, contaminated soils can be restored by using bioremediation techniques, which use plants alone and in association with different beneficial microbes as cutting-edge approaches. This review covers the bioremediation of soils contaminated with Cd in various new ways. The bioremediation capability of bacteria and fungi alone and in combination with plants are studied and analyzed. Microbes, including bacteria, fungi and algae, are reported to have a high tolerance for metals, having a 98% bioremediation capability. The internal structure of microorganisms, their cell surface characteristics and the surrounding environmental circumstances are all discussed concerning how microbes detoxify metals. Moreover, issues affecting the effectiveness of bioremediation are explored, along with potential difficulties, solutions and prospects.

Bacterial ghost cell based bivalent candidate vaccine against Salmonella Typhi and Salmonella Paratyphi A: A prophylactic study in BALB/c mice

Typhoid and emerging paratyphoid fever are a severe enteric disease worldwide with high morbidity and mortality. Licensed typhoid vaccines are in the market, but no paratyphoid vaccine is currently available. In the present study we developed a bivalent vaccine against Salmonella Typhi and Paratyphi A using a bacterial ghost platform. Bacterial ghost cells (BGs) are bacteria-derived cell membranes without cytoplasmic contents that retain their cellular morphology, including all cell surface features. Furthermore, BGs have inherent adjuvant properties that promote an enhanced humoral and cellular immune reaction to the target antigen. Sodium hydroxide was used to prepare ghost cells of Salmonella Typhi and Paratyphi A. The bacterial ghost cells were characterised using electron microscopy. Then BALB/c mice were immunized three times (0th, 14th and 28th day) with the bivalent typhoidal bacterial ghost cells. Haematological study of adult mice throughout immunization period reflected that the immunogen was safe to administer and does not affect the animals’ health. After complete immunization, we found significant serum antibody titter against whole cell lysate, outer membrane protein and lipopolysaccharide of both bacteria, and cell-mediated immunity was observed in an ex-vivo experiment. CD4+, CD8a+ and CD19+ splenic cell populations were increased in immunized animals. Bivalent Typhoidal ghost cell immunized mice showed better survival, less bacterial colonization in systemic organs, and less inflammation and/or destruction of tissue in histopathological analysis than non-immunized control mice.Serum antibodies of immunized animals can significantly inhibit bacterial motility and mucin penetration ability with better killing properties against Salmonella Typhi and Paratyphi A. Furthermore, significant passive protection was observed through the adoptive transfer of serum antibody and lymphocytes of immunized animals to naïve animals after bacterial infection. In summary, Bivalent Typhoidal Bacterial Ghost cells (BTBGs) enhances immunogenic properties and serves as a safe and effective prevention strategy against Salmonella Typhi and Paratyphi A.

Dibenzothiophene removal by environmental bacteria with differential accumulation of intracellular inorganic polyphosphate

Dibenzothiophene (DBT), which belongs to the group of polycyclic aromatic heterocycles of sulfur, is a model substance to study the removal of sulfur compounds from oil due to its recalcitrance to traditional and specific removal processes. The aim of this work was to evaluate DBT bioremoval by environmental bacteria and its relationship with polyphosphate (polyP) accumulation, cell surface characteristics and bioemulsifying activity. Pseudomonas sp. P26 achieved the highest DBT removal percentage (48%) after 7 days of incubation. Moreover, positive correlations were estimated between DBT removal and bioemulsifying activity and biofilm formation. A strain-dependent relationship between the content of intracellular polyP and the presence of DBT in the culture medium was also demonstrated. The study of these bacterial characteristics, which could promote DBT transformation, is a first approach to select DBT-removing bacteria, in order to develop bioformulations that are able to contribute to desulfurization processes of petroleum-derived pollutants in the future.

Biosorption of Azo Dyes by Bacterial Biomass: A review

This review explores the potential of bacterial biomass as a sustainable and cost-effective approach for removing azo dyes from wastewater. Azo dyes, widely used in various industries, pose significant environmental challenges due to their persistence and potential toxic effects. The study provides an extensive analysis of the current literature on the biosorption of dye using bacterial biomass. It discusses the mechanisms involved in biosorption, including physicochemical interactions, microbial metabolism, and cell surface characteristics. The review presents an overview of different bacterial species, their suitability for biosorption, and the factors that influence their efficiency. The review critically evaluates various parameters affecting biosorption performance, such as pH, temperature, initial dye concentration, and biomass dosage. It highlights the importance of optimizing these parameters to enhance biosorption efficacy and maximize dye removal efficiency. The advantages and limitations of using bacterial biomass for azo dye biosorption and comparing it with other conventional treatment methods were discussed. The potential application of biosorption in large-scale scenarios and the challenges associated with its implementation are also addressed. The review emphasizes the need for further studies to explore novel bacterial strains, improve biosorption kinetics, optimize process parameters, and investigate the fate of dye-loaded biomass.

Evaluation cell surface hydrophobicity of four potential probiotics Lactic acid bacteria's isolated from local dairy products

The purpose of this study was to investigate the cell surface hydrophobicity of four Lactic acid bacteria (LAB) strains isolated from various traditional and industry fermented yogurt and cheese sources based on the biochemical tests. The determining cell surface hydrophobicity is a critical step in selecting LAB strains with probiotic qualities and is one of the most essential elements regulating LAB's particular adherence to abiotic surfaces, as well as one of the most critical factors controlling LAB's absorption and destruction of hydrophobic organic matter. In current study, the four LAB strains had high cell surface hydrophobicity to the four separate hydrocarbon solvents xylene, hexadecane, chloroform, ethyl acetate determined via spectrophotometer at 450 nm. The highest hydrophobicity score was determined for LB3 for chloroform, n- hexadecane, ethyl acetate and xylene at 66.51, 38.23, 33.67 and 24.18% respectively. LB3 demonstrated promising cell surface characteristics, suggesting that it could be used as an indigenous probiotic

Unveiling the Morphostructural Plasticity of Zoonotic Sporotrichosis Fungal Strains: Possible Implications for Sporothrix brasiliensis Virulence and Pathogenicity.

Sporotrichosis is a fungal infection caused by Sporothrix species, with Sporothrix brasiliensis as a prevalent pathogen in Latin America. Despite its clinical importance, the virulence factors of S. brasiliensis and their impact on the pathogenesis of sporotrichosis are still poorly understood. This study evaluated the morphostructural plasticity of S. brasiliensis, a fungus that causes sporotrichosis. Three cell surface characteristics, namely cell surface hydrophobicity, Zeta potential, and conductance, were assessed. Biofilm formation was also analyzed, with measurements taken for biomass, extracellular matrix, and metabolic activity. In addition, other potential and poorly studied characteristics correlated with virulence such as lipid bodies, chitin, and cell size were evaluated. The results revealed that the major phenotsypic features associated with fungal virulence in the studied S. brasiliensis strains were chitin, lipid bodies, and conductance. The dendrogram clustered the strains based on their overall similarity in the production of these factors. Correlation analyses showed that hydrophobicity was strongly linked to the production of biomass and extracellular matrix, while there was a weaker association between Zeta potential and size, and lipid bodies and chitin. This study provides valuable insights into the virulence factors of S. brasiliensis and their potential role in the pathogenesis of sporotrichosis.

Probiotic Lactic Acid Bacteria for Vaginal Application. Optimization of Biomass Production and Freeze-Drying Conditions

Probiotic formula for the vaginal tract must contain high numbers of viable beneficial bacteria that maintain their characteristics during the production and further storage, in order to exert the claimed probiotic effect. Four probiotic strains—Lactobacillus gasseri CRL1320, Limosilactobacillus reuteri CRL1324, Ligilactobacillus salivarius CRL1328 and Lacticaseibacillus rhamnosus CRL1332—originally isolated from vagina of healthy women exhibiting beneficial properties were evaluated. Therefore, the optimization of growth parameters and dry biomass production with high viability while preserving probiotic features of strains is a great challenge. Different growth conditions in MRS medium were set up in a laboratory bioreactor; two initial pH and agitation were recorded speeds during static or controlled fermentations. Production conditions of 37 °C with controlled pH 6.5 and 5.5-MRS with 150 and 75 rpm stirring speeds were used for CRL1329 and CRL1332 strains, respectively, while static and free pH MRS for CRL1324 and CRL1328 probiotics allowed us to obtain maximal cell viability counts. However, during 12 h of fermentation, biomass yields of 19.3, 16.2 and 15.2 g/L were achieved when CRL1329, CRL1328 and CRL1324 probiotic strains were grown in static and free pH MRS. The highest biomass yield for CRL1332 strain was produced under controlled MRS-initial pH 6.5 and 75 rpm fermentation conditions. To preserve probiotic high biomass viability, freeze-drying was carried out in the presence of different cryoprotective agents. Thus, the highest viable numbers (10.9–11.8 log CFU/g) with survival rates between 91.3 and 95.6% were attained in the presence of 10% trehalose (L. reuteri and L. salivarius), lactose (L. rhammosus) and lactose + trehalose + sucrose mix (L. gasseri). When stability during post-freeze-drying storage was evaluated, probiotic strains showed a remarkably higher viability recovery when stored at 4 °C than at 25 °C for 12 and 3 months, respectively. In addition, surface characteristics of vaginal probiotics were affected to different extents during storage depending on the strain, protective agent and storage time/temperature. Critical factors for growth conditions, drying process and storage stability of probiotic lactobacilli strains were optimized in view to preserve cell high viability and surface features for the design of vaginal probiotic formula.