Bacterial Strains Research Articles

High-throughput method for screening pendimethalin-degrading bacteria from one microbial bank.

The extensive use of chemical pesticides, such as herbicides, has resulted in significant environmental pollution. Microbial degradation represents a crucial approach for managing this pesticide-associated pollution, with enrichment culturing serving as a method for isolating pesticide-degrading microorganisms. However, the efficiency of this strategy is limited, often yielding only a few isolated strains. In this study, a new mineral salt medium (MSM) was developed, and a high-throughput method was used for screening pendimethalin-degrading bacteria by measuring the bacterial growth in the MSM. The utilization of this method resulted in the isolation of 56 pendimethalin-degrading bacteria from approximately 2000 bacterial strains, including 37 Bacillus spp., 10 Alcaligenes spp., 5 Pseudomonas spp., and other 4 strains identified for the first time as pendimethalin-degrading strains. This method may hold promise not only for isolating bacterial strains capable of degrading other pesticides but also for facilitating the utilization of the substantial bacterial strains stored in bacterial banks.

Isolation and Molecular Characterization of Polyester-Degrading Bacterial Strains from Municipal Wastewater and Their Application in Microplastic Degradation in Contaminated Wastewater

Diverse synthetic polymers find their regular application in routine life and are disposed of directly into the environment through various sources. The released microplastic and microfibres from the sources creates contamination in both aquatic and terrestrial environments. When these micropollutants enter the food chain, they have adverse effects on the environment and human health. This study focuses on the application of native bacterial strains isolated for the effective degradation of polyester microfibers. The isolated bacterial strains, namely MFB 6, MFB 8, and MFB 11, tolerant to polyethylene glycol (PEG), were identified as Bacillus sp., Paenibacillus sp., and Aeromonas veronii, respectively, with NCBI gene bank accession numbers ON318306, ON318390, and PP237260. Polyester biodegradation efficiency of 10.5%, 12.6% and 8.4% was achieved by MFB 6, MFB 8 and MFB 11, respectively, after 42 days of investigation in minimal salt medium under shake flask condition. In future the application of these microorganisms will help in sustainable management of synthetic microfiber pollutants and find their suitable application in micro plastic degradation in contaminated wastewater.

Anticandidal applications of selenium nanoparticles biosynthesized with Limosilactobacillus fermentum (OR553490).

Candida albicans is one of the most dangerous pathogenic fungi in the world, according to the classification of the World Health Organization, due to the continued development of its resistance to currently available anticandidal agents. To overcome this problem, the current work provided a simple, one-step, cost-effective, and safe technique for the biosynthesis of new functionalized anticandidal selenium nanoparticles (Se NPs) against C. albicans ATCC10231 using the cell-free supernatant of Limosilactobacillus fermentum (OR553490) strain. The bacterial strain was isolated from yogurt samples available in supermarkets, in Damietta, Egypt. The mixing ratio of 1:9v/v% between cell-free bacterial metabolites and sodium selenite (5 mM) for 72 h at 37°C were the optimum conditions for Se NPs biosynthesis. Ultraviolet-visible spectroscopy (UV-Vis), Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray diffraction (XRD), Zeta analyses, and elemental analysis system (EDS) were used to evaluate the optimized Se NPs. The Se NPs absorption peak appeared at 254 nm. Physicochemical analysis of Se NPs revealed the crystalline-shaped and well-dispersed formation of NPs with an average particle size of 17-30 nm. Se NPs have -11.8mV, as seen by the zeta potential graph. FT-IR spectrum displayed bands of symmetric and asymmetric amines at 3279.36 cm-1 and 2928.38 cm-1, aromatic and aliphatic (C-N) at 1393.32 cm-1 and 1237.11.37 cm-1 confirming the presence of proteins as stabilizing and capping agents. Se NPs acted as a superior inhibitor of C. albicans with an inhibition zone of 26 ± 0.03mm and MIC value of 15 µg/mL compared to one of the traditional anticandidal agent, miconazole, which revealed 18 ± 0.14mm and 75µg/mL. The cytotoxicity test shows that Se NPs have a low toxic effect on the normal keratinocyte (IC50 ≈ 41.5 μg/mL). The results indicate that this green synthesis of Se NPs may have a promising potential to provide a new strategy for drug therapy.

Production of Poly(3-Hydroxybutyrate-Co-3-Hydroxyvalerate) by Bacillus megaterium LVN01 Using Biogas Digestate

The Bacillus megaterium LVN01 species native to Colombia has demonstrated the ability to metabolize different coproducts or industrial waste (such as fique juice, cane molasses, and residual glycerol) and accumulate polyhydroxybutyrate (PHB), giving it potential in the bioplastics industry. In this research, the potential of liquid digestate as a carbon source for the production of PHA polymers in fermentation processes with this bacterial strain was evaluated. Favorably, it was found that B. megaterium utilizes the nutrients from this residual substrate to multiply appropriately and efficiently synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). Bench-scale aerobic batch fermentation, under the operational conditions of this research [volume: 3 L; temperature: 30.8 °C; agitation: 400 rpm; pH: 7.0 ± 0.2; dissolved oxygen: 100% saturation; antifoam: 10% (v/v)], generated maximum values of dry cell weight (DCW) (0.56 g cell L−1) at 60 h, while the maximum PHBV yield (360 mg PHBV L−1) occurred at 16 h, which is very favorable for sustainable degradable bioplastics production. Additionally, GC–MS and NMR analyses confirmed that the PHBV copolymer synthesized by B. megaterium is made up of the monomers 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV). Furthermore, the thermal properties determined by TGA (Tonset = 283.1 °C; Tendset = 296.98 °C; Td = 290.114 °C) and DSC (Tm = °C 155.7 °C; ΔHf = 19.80 J g−1; Xcr = 18.17%) indicate that it is a thermally stable biopolymer with low percentages of crystallinity, providing flexibility that facilitates molding, adaptation, and application in various industrial sectors.

Production of Poly(3-Hydroxybutyrate-Co-3-Hydroxyvalerate) by Bacillus megaterium LVN01 Using Biogas Digestate

The Bacillus megaterium LVN01 species native to Colombia has demonstrated the ability to metabolize different coproducts or industrial waste (such as fique juice, cane molasses, and residual glycerol) and accumulate polyhydroxybutyrate (PHB), giving it potential in the bioplastics industry. In this research, the potential of liquid digestate as a carbon source for the production of PHA polymers in fermentation processes with this bacterial strain was evaluated. Favorably, it was found that B. megaterium utilizes the nutrients from this residual substrate to multiply appropriately and efficiently synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). Bench-scale aerobic batch fermentation, under the operational conditions of this research [volume: 3 L; temperature: 30.8 °C; agitation: 400 rpm; pH: 7.0 ± 0.2; dissolved oxygen: 100% saturation; antifoam: 10% (v/v)], generated maximum values of dry cell weight (DCW) (0.56 g cell L−1) at 60 h, while the maximum PHBV yield (360 mg PHBV L−1) occurred at 16 h, which is very favorable for sustainable degradable bioplastics production. Additionally, GC–MS and NMR analyses confirmed that the PHBV copolymer synthesized by B. megaterium is made up of the monomers 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV). Furthermore, the thermal properties determined by TGA (Tonset = 283.1 °C; Tendset = 296.98 °C; Td = 290.114 °C) and DSC (Tm = °C 155.7 °C; ΔHf = 19.80 J g−1; Xcr = 18.17%) indicate that it is a thermally stable biopolymer with low percentages of crystallinity, providing flexibility that facilitates molding, adaptation, and application in various industrial sectors.

Nano Zinc Oxide Particle Synthesis from Bio-Waste Selaginella willdenowii Leaf Extract: A Multi-Faceted Approach for Environmental and Biomedical Applications

Selaginella willdenowii, a commonly used greenhouse fern, was often used as a biowaste to synthesize zinc oxide nanoparticles (ZnO NPs) in an eco-friendly and cost-effective way. UV–Visible spectra studies were carried out to confirm the synthesis of S. willdenowii-mediated ZnO NPs (SW-ZnO NPs), and a peak at 367[Formula: see text]nm with a sharp band gap of 3.415[Formula: see text]eV was observed. The X-ray diffraction analysis indicated that the crystalline size of the synthesized SW-ZnO NPs was 11.971[Formula: see text]nm. The phytochemicals present in the extracts and the compounds involved in the reduction of metal to nanoparticles were determined by Fourier Transform Infrared analysis. Scanning electron microscopy was utilized to analyze the surface morphology and size of the obtained SW-ZnO NPs. The examination revealed that they exhibited a hexagonal shape, with an average size falling within the range of 17–23[Formula: see text]nm. Under ultra-violet light, reactive blue 220 and reactive yellow 145 dyes showed 78.06% and 60.14% degradation, showing potential photocatalytic degradation activity. The synthesized SW-ZnO NPs also exhibited antimicrobial activity against bacterial strains (Escherichia coli and Bacillus subtilis) and fungal cultures (Candida tropicalis and Candida albicans) showed cytotoxic activity against Hep-G2 cell lines. Our results suggest the green synthesized SW-ZnO NPs have potential photocatalytic, antimicrobial and cytotoxic potential.

Microwave-assisted synthesis of novel pyrazolyl sulfones and their antimicrobial evaluation and time-resolved photoluminescence studies

This article reports an efficient and practical microwave-assisted MMPP-promoted synthesis of novel pyrazolyl sulfones from the corresponding 4-(alkyl/cycloalkylthio)-1H-pyrazoles. The reaction of 4-(alkyl/cycloalkylthio)-1H-pyrazoles with magnesium bis(monoperoxyphthalate)hexahydrate (MMPP) as an oxidizing agent afforded the corresponding 4-(alkyl/cycloalkylsulfonyl)-1,3-disubstituted-1H-pyrazoles in 85–95% yield. This method's benefits include its straightforward operation, simple workup, and use of an inexpensive, halogen-free MMPP oxidant that is easy to use and reasonably stable. All synthesized pyrazolyl sulfones were examined against bacterial and fungal strains, and notable antimicrobial activity was demonstrated by a few of the compounds. In order to investigate the prospect of connecting compounds with the highest yield and antimicrobial activity with their opto-electronic properties, time-resolved photoluminescence investigations for compounds 5a, 5b, and 5d were conducted. The tunable spectrum was observed in the micro-second time domain in all three cases, with fluorescent lifetime found higher in the compound having methyl group, intermediate with phenyl group, and lowest with the p-nitrophenyl group. Our procedures will encourage additional research into the valuable properties of pyrazolyl sulfones now that they are widely available with the structural complexity illustrated here.

Photodynamic Inactivation as a Promising Method of Combating Resistant Strains of Staphylococci

Relevance. The development of antimicrobial drugs and alternative methods, technologies and means of prevention, diagnosis and treatment of human infectious diseases caused by antibiotic-resistant microorganisms is one of the priorities of ensuring the biological safety of the country. Aims. To evaluate the bactericidal activity of tetrapyrrole macroheterocycles (porphyrins) at different light irradiation durations in relation to staphylococci, in vitro. Materials and methods. Studied strains of microorganisms: museum strains of microorganisms – S. aureus ATCC 29213, S. epidermidis ATCC 14990 and antibiotic-resistant strains of bacteria of the genus Staphylococcus (n=18) isolated from clinical biomaterial and from environmental objects in a medical organization. The studied chemical compounds are three different compounds of water-soluble asymmetrically substituted porphyrins containing heterocyclic fragments on the periphery of the porphyrin cycle (residues of benzoxazole, N-methylbenzimidazole and benzothiazole). Results. The activity of all three porphyrin compounds in relation to museum strains of staphylococcus and 77.8% of clinical antibiotic-resistant strains (n=14; 95% CI 20.1-97.5) turned out to be maximal (complete lysis) after 10 minutes of irradiation. Conclusions. The tested tetrapyrrole macroheterocycles (porphyrins) exhibit bactericidal activity against museum and clinical strains of staphylococcus, with different levels of antibiotic resistance, which determines Keywords: antibiotic resistance, water-soluble porphyrin, photodynamic inactivation, photosensitizer, photochemistry, staphylococci No conflict of interest to declare.

Potential of the plant growth-promoting rhizobacterium Rhodococcus qingshengii LMR356 in mitigating lead stress impact on Sulla spinosissima L.

Mining-related lead (Pb) pollution of the soil poses serious hazards to ecosystems and living organisms, including humans. Improved heavy metal phytoremediation efficacy, achieved by using phytostabilizing plants assisted by plant-growth-promoting (PGP) microorganisms, has been presented as an effective strategy for remediating polluted soils. The objective of this research was to examine the response and potential of the plant-growth-promoting bacterium LMR356, a Rhodococcus qingshengii strain isolated from an abandoned mining soil, under lead stress conditions. Compared to non-contaminated culture media, the presence of lead induced a significant decrease in auxin production (from 21.17 to 2.65 μg mL-1) and phosphate solubilization (from 33.60 to 8.22 mg L-1), whereas other PGP traits increased drastically, such as 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity (from 38.17 to 71.37 nmol mg-1 h-1 α-ketobutyrate), siderophore production (from 69 to 83%), exopolysaccharide production (from 1952.28 to 3637.72 mg mL-1), biofilm formation, and motility. We, therefore, investigated the behavior of Sulla spinosissima L. in the presence or absence of this strain under a variety of experimental conditions. Under hydroponic conditions, Sulla plants showed endurance to varying lead concentrations (500-1000 μM). Inoculation of plants with Rhodococcus qingshengii strain LMR356 enhanced plant tolerance, as demonstrated by the increase in plant biomass (ranging from 14.41 to 79.12%) compared to non-inoculated Pb-stressed and non-stressed control plants. Antioxidant enzyme activities (increasing by -42.71 to 126.8%) and chlorophyll (383.33%) and carotenoid (613.04%) content were also augmented. In addition to its impact on plant lead tolerance, strain LMR356 showed a growth-promoting effect on Sulla plants when cultivated in sterilized non-contaminated sand. Parameters such as plant biomass (16.57%), chlorophyll (24.14%), and carotenoid (30%) contents, as well as ascorbate peroxidase (APX), peroxidase (POD), and catalase (CAT) activities, were all elevated compared to non-inoculated plants. Furthermore, when the same plant species was cultivated in highly polluted soil, inoculation increased plant biomass and improved its physiological properties. These findings demonstrate that LMR356 is a phytobeneficial bacterial strain capable of enhancing Sulla growth under normal conditions and improving its heavy metal tolerance in multi-polluted soils. Thus, it can be considered a promising biofertilizer candidate for growing Sulla spinosissima L. or other selected plants intended for application in restoration and stabilization initiatives aimed at reviving and safeguarding environmentally compromised and polluted soils after mining activities.

Current challenges and improvements in assessing the immunogenicity of bacterial vaccines

The increase in antimicrobial-resistant bacterial strains has highlighted the need for a new vaccine strategy. The primary goal of a candidate vaccine is to prevent disease, by inducing a persistent immunologic memory, through the activation of pathogen-specific immune response. Antibody titer is the main parameter used to assess the immunogenicity of bacterial vaccine candidates and it is the most widely used as a correlate of protection. On the other hand, the antibody titer alone cannot provide complete information on all the activity mediated by antibodies which can only be assessed by functional assays, like the serum bactericidal assay and the opsonophagocytosis assay. However, due to the involvement of many biological factors, these assays are difficult to standardize. Some improvements have been achieved in recent years, but further optimizations are needed to minimize inter- and intra-laboratories variability and to allow the applicability of these functional assays for the vaccine immunogenicity assessment on a larger scale.

A novel cost-effective methodology for the screening of nanocellulose producing micro-organisms.

In this paper, the work has been done to develop a cost-effective methodology, for the isolation of the potential producer of bacterial nanocellulose. No report is available in the literature, on the use of gram flour and table sugar for the screening of nanocellulose-producing isolates. Since commercially used, Hestrin-Schramm medium is expensive for the isolation of nanocellulose-producing micro-organisms, the possibility of using gram flour-table sugar medium was investigated in this work. Qualitative screening of micro-organisms was done using cost-effective medium, i.e., gram flour-table sugar medium. Qualitative analysis of various nanocellulose-producing bacteria depicted that cellulose layer production occurred on both HS medium and gram flour-table sugar medium. The yield of nanocellulose was also better on air-liquid surface in case of gram flour-table sugar medium as compared to HS medium. 16S rRNA was used for molecular characterization of bacterial strain and the best nanocellulose producer was identified as Novacetimonas hansenii BMK-3_NC240423 (isolated from rotten banana). FTIR and FE-SEM studies of nanocellulose pellicle produced on HS medium and gram flour-table sugar medium demonstrated equivalent structural, morphological, and chemical properties. The cost of newly designed medium (0.01967$/L) is nearly 90 times lower than the Hestrin-Schramm medium (1.748$/L), which makes the screening of nanocellulose producers very cost-effective. A strategy of using gram flour extract-table sugar medium for the screening of nanocellulose-producing micro-organisms is a novel approach, which will drastically reduce the screening associated cost of cellulose-producing micro-organisms and also motivate the researchers/industries for comprehensive screening programme for getting high cellulose-producing microbes.

Current challenges and improvements in assessing the immunogenicity of bacterial vaccines

The increase in antimicrobial-resistant bacterial strains has highlighted the need for a new vaccine strategy. The primary goal of a candidate vaccine is to prevent disease, by inducing a persistent immunologic memory, through the activation of pathogen-specific immune response. Antibody titer is the main parameter used to assess the immunogenicity of bacterial vaccine candidates and it is the most widely used as a correlate of protection. On the other hand, the antibody titer alone cannot provide complete information on all the activity mediated by antibodies which can only be assessed by functional assays, like the serum bactericidal assay and the opsonophagocytosis assay. However, due to the involvement of many biological factors, these assays are difficult to standardize. Some improvements have been achieved in recent years, but further optimizations are needed to minimize inter- and intra-laboratories variability and to allow the applicability of these functional assays for the vaccine immunogenicity assessment on a larger scale.

Enterococcus spp. in fish: Analysis of the presence and resistance in samples from Tri-City, Poland.

The growing concern over antibiotic resistance in foodborne pathogens necessitates comprehensive assessments of its prevalence and associated risks in various food products. The present study aimed to assess the occurrence of Enterococcus spp. in samples of fish purchased at various points of sale in the Tricity region. The selection of products (n = 74) was based on their availability and included both fish caught in the Baltic region and products imported from, Vietnam, China, Norway, and European Union (EU) countries. For bacterial isolation, samples were inoculated into selective broth, and the growth of enterococci was assessed based on turbidity. Positive cultures were confirmed by a change in color in bromocresol purple broth and were isolated on Slanetz-Bartley agar. Bacteria were present in all tested samples regardless of the degree of raw material processing as follows: frozen (F)- 55%, fresh/raw (FS)- 70.6%, thawed (DF)- 30%, smoked (S)- 50%, and the packaging methods, modified atmosphere packaging (MAP)- 34.4%, unit packaging (UP)- 75%, and sold in bulk (SB)- 76.9%, with an overall frequency of occurrence of 58.1%. The number of bacteria ranged from not detected to 4.28-log cfu/g, with the lowest mean values for thawed fish and those packed in MAP. Tests conducted on 24 strains isolated from samples showed their varied sensitivity to tetracyclines. Single cases of multidrug resistance of the tested strains were also observed. The conducted statistical analysis did not show statistically significant differences in the count of enterococci based on the origin, degree of processing, or packaging (p < 0.05). Moreover, differences in strain sensitivity to ampicillin were observed. Detected cases of resistance, especially to tetracycline, require careful monitoring and action to limit the health risks associated with resistant bacterial strains in food products.

Antibacterial Activity of Brass against Antibiotic-Resistant Bacteria following Repeated Exposure to Hydrogen Peroxide/Peracetic Acid and Quaternary Ammonium Compounds

Copper-containing materials are attracting attention as self-disinfecting surfaces, suitable for helping healthcare settings in reducing healthcare-associated infections. However, the impact of repeated exposure to disinfectants frequently used in biocleaning protocols on their antibacterial activity remains insufficiently characterized. This study aimed at evaluating the antibacterial efficiency of copper (positive control), a brass alloy (AB+®) and stainless steel (negative control) after repeated exposure to a quaternary ammonium compound and/or a mix of peracetic acid/hydrogen peroxide routinely used in healthcare settings. A panel of six antibiotic-resistant strains (clinical isolates) was selected for this assessment. After a short (5 min) exposure time, the copper and brass materials retained significantly better antibacterial efficiencies than stainless steel, regardless of the bacterial strain or disinfectant treatment considered. Moreover, post treatment with both disinfectant products, copper-containing materials still reached similar levels of antibacterial efficiency to those obtained before treatment. Antibiotic resistance mechanisms such as efflux pump overexpression did not impair the antibacterial efficiency of copper-containing materials, nor did the presence of one or several genes related to copper homeostasis/resistance. In light of these results, surfaces made out of copper and brass remain interesting tools in the fight against the dissemination of antibiotic-resistant strains that might cause healthcare-associated infections.

Nano-Bioremediation of Arsenic and Its Effect on the Biological Activity and Growth of Maize Plants Grown in Highly Arsenic-Contaminated Soil.

Arsenic (As)-contaminated soil reduces soil quality and leads to soil degradation, and traditional remediation strategies are expensive or typically produce hazardous by-products that have negative impacts on ecosystems. Therefore, this investigation attempts to assess the impact of As-tolerant bacterial isolates via a bacterial Rhizobim nepotum strain (B1), a bacterial Glutamicibacter halophytocola strain (B2), and MgO-NPs (N) and their combinations on the arsenic content, biological activity, and growth characteristics of maize plants cultivated in highly As-contaminated soil (300 mg As Kg-1). The results indicated that the spectroscopic characterization of MgO-NPs contained functional groups (e.g., Mg-O, OH, and Si-O-Si) and possessed a large surface area. Under As stress, its addition boosted the growth of plants, biomass, and chlorophyll levels while decreasing As uptake. Co-inoculation of R. nepotum and G. halophytocola had the highest significant values for chlorophyll content, soil organic matter (SOM), microbial biomass (MBC), dehydrogenase activity (DHA), and total number of bacteria compared to other treatments, which played an essential role in increasing maize growth. The addition of R. nepotum and G. halophytocola alone or in combination with MgO-NPs significantly decreased As uptake and increased the biological activity and growth characteristics of maize plants cultivated in highly arsenic-contaminated soil. Considering the results of this investigation, the combination of G. halophytocola with MgO-NPs can be used as a nanobioremediation strategy for remediating severely arsenic-contaminated soil and also improving the biological activity and growth parameters of maize plants.

Phytotoxicity, cytogenotoxicity and antimicrobial potential of extracts with gold-silver bimetallic nanoparticles obtained from pteridophyte spores

Investigating the toxicity of naturally occurring or synthesized nanoparticles for various applications is absolutely necessary for environmental protection and safety use. The aim of these research was to investigated the phytotoxicity, cytogenotoxicity and antibacterial potential of the extracts with gold-silver bimetallic nanoparticles (Au-Ag NPs) obtained from green synthesis in Asplenium scolopendrium L. and Dryopteris filix-mas (L.) Schott spores extracts. To our knowledge, this is the first report of the Au-Ag NPs phytosynthesis based on extracts obtained from fern spores. UV-Vis spectroscopy analysis of the samples revealed the maximum absorbance, characteristic of samples with bimetallic nanoparticles, which varied depending on the Au:Ag ratio. Energy-dispersive X-ray spectroscopy confirmed the presence and distribution of Au, Ag and other chemical elements. The presence of specific secondary metabolites in the extracts that helped in NPs biosynthesis stimulated growth processes. Good results were recorded for some Dryopteris filix-mas samples, correlated with a significantly increased mitotic index. Cell viability decreased significantly in three of the nanoformulations. Only extracts with Au-Ag NPs showed antimicrobial effect against Staphylococcus aureus ATCC 25923, Bacillus subtilis ATCC 6633 and Escherichia coli ATCC 8739. The testing of the antibacterial potential of these extracts must be extended to other bacterial strains and other microorganisms, the search of new antimicrobial resources being an urgent necessity nowadays.

Phytotoxicity, cytogenotoxicity and antimicrobial potential of extracts with gold-silver bimetallic nanoparticles obtained from pteridophyte spores

Investigating the toxicity of naturally occurring or synthesized nanoparticles for various applications is absolutely necessary for environmental protection and safety use. The aim of these research was to investigated the phytotoxicity, cytogenotoxicity and antibacterial potential of the extracts with gold-silver bimetallic nanoparticles (Au-Ag NPs) obtained from green synthesis in Asplenium scolopendrium L. and Dryopteris filix-mas (L.) Schott spores extracts. To our knowledge, this is the first report of the Au-Ag NPs phytosynthesis based on extracts obtained from fern spores. UV-Vis spectroscopy analysis of the samples revealed the maximum absorbance, characteristic of samples with bimetallic nanoparticles, which varied depending on the Au:Ag ratio. Energy-dispersive X-ray spectroscopy confirmed the presence and distribution of Au, Ag and other chemical elements. The presence of specific secondary metabolites in the extracts that helped in NPs biosynthesis stimulated growth processes. Good results were recorded for some Dryopteris filix-mas samples, correlated with a significantly increased mitotic index. Cell viability decreased significantly in three of the nanoformulations. Only extracts with Au-Ag NPs showed antimicrobial effect against Staphylococcus aureus ATCC 25923, Bacillus subtilis ATCC 6633 and Escherichia coli ATCC 8739. The testing of the antibacterial potential of these extracts must be extended to other bacterial strains and other microorganisms, the search of new antimicrobial resources being an urgent necessity nowadays.

Synergistic effect of partially hydrolyzed guar gum on Clostridium butyricum in a synbiotic combination for enhanced butyrate production during in-vitro fermentation

Background: Clostridium butyricum is a butyrate-producing beneficial bacterium and is generally recognized as a significant indicator of appropriate gut microbial metabolism in human health. Objective: The synergistic effects of commercially available prebiotic partially hydrolyzed guar gum (PHGG) dietary fiber as a carbon source on a butyrate-producing bacterial strain were evaluated during in-vitro fermentation with Clostridium butyricum bacterial strain in a basal medium. Additionally, their prebiotic activities were compared to those of other dietary fibers. Methods: The examined functional dietary fiber substrates (PHGG, LMW-PHGG, indigestible dextrin, and inulin) demonstrated selective prebiotic effects on pH variation of a basal medium, leading to enhanced bacterial growth and butyrate production with Clostridium butyricum bacterial strains during in-vitro fermentation. Results: Prebiotic PHGG supplementation had the highest fermentability among dietary fibers, resulting in greater bacterial growth (OD660: 1.93 ± 0.01) of the Clostridium butyricum strain and enhanced butyrate generation (4.52 ± 2.09 mM) after cultivation in a basal medium. A significant difference in promoting bacterial growth (p &lt; 0.05), pH reduction (p &lt; 0.05), and butyrate production (p &lt; 0.05) compared to indigestible dextrin and inulin was observed. Mannose demonstrated the strongest butyrogenic effect and improved fermentability on the Clostridium butyricum, among the studied prebiotic monosaccharides (galactose, glucose, and starch). The order of bacterial growth and butyrate synthesis was mannose &gt; galactose &gt; glucose &gt; starch. The PHGG with a relatively lower molecular weight (LMW-PHGG) exhibited the improved bacterial growth of Clostridium butyricum and demonstrated the highest butyrate production after cultivation in a basal medium. A similar trend was observed when Clostridium butyricum was cultivated in-vitro using PHGG-supplemented artificial intestinal fluid containing MRS-agar medium. Conclusion: These findings suggest that the symbiotic combination of prebiotic PHGG and probiotic Clostridium butyricum could have major industrial applications as a therapeutic adjuvant for improved gastrointestinal health. Keywords: Partially hydrolyzed guar gum, synbiotic, prebiotic, probiotic, Clostridium butyricum, butyrate production

Preliminary Identification and Quantification of Individual Polyphenols in Fallopia japonica Plants and Honey and Their Influence on Antimicrobial and Antibiofilm Activities.

Fallopia japonica (FJ), an invasive plant species known for its rich bioactive compounds, has been used for centuries in traditional Chinese medicine. Despite its significant beekeeping potential, this aspect of FJ remains underexplored. This research aims to investigate the antimicrobial and antibiofilm properties of FJ plants and honey. Notably, this study is the first to identify individual phenolic compounds in both FJ plant tissues and FJ honey, highlighting resveratrol as a marker of FJ honey. The study tested inhibitory activity against seven bacterial strains: Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, Salmonella enteritidis, and the yeast Candida albicans. Disk diffusion and microdilution methods were used to assess antimicrobial activity, while the crystal violet staining test evaluated antibiofilm activity. Results showed that FJ plant tissues and honey exhibited strong inhibition, particularly against Gram-negative bacterial strains. The most significant inhibition of biofilm formation, by both FJ plant tissues and honey, was observed against Staphylococcus aureus and Escherichia coli. A significant positive correlation was found between antimicrobial activity and individual polyphenols, especially resveratrol. The antibacterial and antibiofilm potential of FJ plant tissues and honey suggests promising applications in sustainable beekeeping. Further research is necessary to evaluate the bioactive compounds found in FJ honey and their health effects.

Microbial influence on the larval survival of Japanese eel Anguilla japonica: Antibiotic-mediated alterations and biomarker isolation.

In rearing systems for the Japanese eel Anguilla japonica, although it is assumed that microorganisms influence larval survival and mortality, particularly during the early stages of growth, the effects of bacterial communities on larval survival have yet to be sufficiently determined. In this study, we compared the bacterial communities associated with larval survival at three stages of eel growth. To artificially alter bacterial communities and assess larval survival, eel larvae were treated with 11 types of antibiotic, and larval survival and bacterial characteristics were compared between the antibiotic-treated and antibiotic-free control groups. Throughout the three growth stages, eels treated with four antibiotics (polymyxin B, tetracycline, novobiocin, and erythromycin) had survival rates higher than those in the control groups. The bacterial communities of surviving larvae in the control and antibiotic groups and dead larvae in the control groups were subsequently analyzed using 16S rRNA gene amplicon sequencing. PERMANOVA analysis indicated that these three larval groups were characterized by significantly different bacterial communities. We identified 14 biomarker amplicon sequence variants (ASVs) of bacterial genera such as Oceanobacter, Alcanivorax, Marinobacter, Roseibium, and Sneathiella that were enriched in surviving larvae in the antibiotic treatment groups. In contrast, all four biomarker ASVs enriched in dead larvae of the control groups were from bacteria in the genus Vibrio. Moreover, 52 bacterial strains corresponding to nine biomarkers were isolated using a culture method. To the best of our knowledge, this is the first study to evaluate the bacterial communities associated with the survival and mortality of larvae in during the early stages of Japanese eel growth and to isolate biomarker bacterial strains. These findings will provide valuable insights for enhancing larval survival in the eel larval rearing systems from a microbiological perspective.