International Microbiology最新文献

This research examined acetic acid bacteria (AAB) isolated from Kombucha beverages produced with Anatolian hawthorn (Crataegus orientalis) as next-generation probiotics. Eighty-six AAB were isolated from the samples and investigated in terms of biosafety, viability in vitro gastrointestinal conditions, technological and bioactive properties, and also in vitro adhesion abilities. Seventy-six isolates demonstrating γ-hemolysis exhibited resistance to erythromycin and ampicillin. Besides, these isolates survived at low pH and in the presence of bile salts. However, the majority of AAB isolates showed tolerance to phenol, pepsin, and pancreatin. Also, twenty-one isolates showed protease enzyme activity, while eight isolates had amylase enzyme activity. Despite most of the isolates showed viability at 1.5% salt, only 19 isolates survived at 10% salt. Most AAB isolates exhibited inhibition zones ranging from 8 to 26 mm against test bacteria, their antioxidant activities were above 80%. Additionally, some isolates exhibited auto-aggregation ability ranging from 0.66 to 23.62% and co-aggregation ability ranging from 1.18 to 71.32%, while hydrophobicity ranged from 1.32 to 69.87% toward xylene. Finally, the indigenous 76 AAB isolates that had remarkable probiotic properties were characterized based on 16S rRNA gene sequencing, and the isolates belonged to Komagateibacter sp. (64.47%), Komagateibacter saccharivorans (15.79%), K. rhaeticus (11.84%), and Gluconobacter sp. (7.90%). As a result, the isolates identified as Gluconobacter sp. A21, Komagataeibacter sp. A139, Gluconobacter sp. A141, and Komagataeibacter sp. A146, which showed high viability under gastrointestinal conditions, safe and acceptable in terms of technological, bioactive, and adhesion properties and could be evaluated as next-generation probiotics.

A new thermophilic strain, designated as Bacillus sp. LMB3902, was isolated from Hammam Debagh, the hottest spring in Algeria (up to 98 °C). This isolate showed high protease production in skim milk media at 55 °C and exhibited significant specific protease activity by using azocasein as a substrate (157.50 U/mg). Through conventional methods, chemotaxonomic characteristics, 16S rRNA gene sequencing, and comparative genomic analysis with the closely related strain Bacillus licheniformis DSM 13 (ATCC 14580 ^T), the isolate Bacillus sp. LMB3902 was identified as a potentially new strain of Bacillus licheniformis. In addition, the gene functions of Bacillus sp. LMB3902 strain were predicted using the Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, Clusters of Orthologous Groups, Non-Redundant Protein Sequence Database, Swiss-Prot, and Pfam databases. The results showed that the genome size of Bacillus sp. LMB3902 was 4.279.557 bp, with an average GC content of 46%. The genome contained 4.760 predicted genes, including 8 rRNAs, 78 tRNAs, and 24 sRNAs. A total of 235 protease genes were annotated including 50 proteases with transmembrane helix structures and eight secreted proteases with signal peptides. Additionally, the majority of secondary metabolites found by antiSMASH platform showed low similarity to identified natural products, such as fengicin (53%), lichenysin (57%), and surfactin (34%), suggesting that this strain may encode for novel uncharacterized natural products which can be useful for biotechnological applications. This study is the first report that describes the complete genome sequence, taxono-genomics, and gene annotation as well as protease production of the Bacillus genus in this hydrothermal vent.

Arbuscular mycorrhizal fungi (AMF) are obligate symbionts that engage in crucial interactions with plants, playing a vital role in grassland ecology. Our study focuses on the pioneer plant Agropyron cristatum, and we collected soil samples from four degraded grasslands in Yudaokou to investigate the response of community composition to the succession of degraded grasslands. We measured the vegetation status, soil physical and chemical properties, AMF colonization, and spore density in different degraded grasslands. High-throughput sequencing was employed to analyze AMF in soil samples. Correlations among community composition, soil characteristics, and plant factors were studied using principal component and regression analyses. The distribution of AMF in grasslands exhibited variation with different degrees of degradation, with Glomus, Scutellospora, and Diversispora being the dominant genera. The abundance of dominant genera in AMF also varied, showing a gradual increase in the relative abundance of the genus Diversispora with higher degradation levels. AMF diversity decreased from 27.7% to 12.4% throughout the degradation process. Among 180 samples of Agropyron cristatum plants, AMF hyphae and vesicles displayed the highest infection status in non-degraded grasslands and the lowest in severely degraded ones. Peak AMF spore production occurred in August, with maximum values in the 0-10-cm soil layer, and the highest spore densities were found in lightly degraded grasslands. Apart from pH, soil factors exhibited a positive correlation with AMF infection during grassland degradation. Furthermore, changes in AMF community composition were jointly driven by vegetation and soil characteristics, with vegetation coverage and soil organic carbon significantly impacting AMF distribution. Significant differences in AMF variables (spore number and diversity index) were also observed at different soil depths. Grassland successional degradation significantly influences AMF community structure and composition. Our future focus will be on understanding response mechanisms and implementing improvement methods for AMF during grassland degradation and subsequent restoration efforts.

Around 1.5 million mortality cases due to fungal infection are reported annually, posing a massive threat to global health. However, the effectiveness of current antifungal therapies in the treatment of invasive fungal infections is limited. Repurposing existing antifungal drugs is an advisable alternative approach for enhancing their effectiveness. This study evaluated the antifungal efficacy of the antiviral drug vidarabine against Candida albicans ATCC 90028. Antifungal susceptibility testing was performed by microbroth dilution assay and further processed to find the minimum fungicidal concentration. Investigation on probable mode of vidarabine action against C. albicans was assessed by using the ergosterol reduction assay, reactive oxygen species (ROS) accumulation, nuclear condensation, and apoptosis assay. Results revealed that C. albicans was susceptible to vidarabine action and exhibited minimum inhibitory concentration at 150 µg/ml. At a concentration of 300 µg/ml, vidarabine had fungicidal activity against C. albicans. 300 µg/ml vidarabine-treated C. albicans cells demonstrated 91% reduced ergosterol content. Annexin/FITC/PI assay showed that vidarabine (150 µg/ml) had increased late apoptotic cells up to 31%. As per the fractional inhibitory concentration index, vidarabine had synergistic activity with fluconazole and caspofungin against this fungus. The mechanism underlying fungicidal action of vidarabine was evaluated at the intracellular level, and probably because of increased nuclear condensation, enhanced ROS generation, and cell cycle arrest. In conclusion, this data is the first to report that vidarabine has potential to be used as a repurposed antifungal agent alone or in combination with standard antifungal drugs, and could be a quick and safe addition to existing therapies for treating fungal infections.

Ectomycorrhizas are ubiquitous symbiotic associations between host trees and soil fungi. While the seasonal changes of the taxonomic community structure of ectomycorrhizal fungi have been studied extensively, the temporal dynamics of ectomycorrhizal exploration types which have been proposed for elucidating the functional roles of ectomycorrhizas have not been fully examined. The purpose of the study is to test the hypothesis of whether the abundance of the exploration types in the hosts with different phenology shows different temporal patterns over the seasons. Two host species, deciduous Quercus acutissima and evergreen Q. glauca, were planted in growth containers with natural forest soils and were grown in single or combined species treatment, under similar environmental conditions and in shared soil spore banks of the ectomycorrhizal fungi. The ectomycorrhizal exploration types that occurred on these two host species in two different treatments were observed for two growing seasons. The observed exploration types, namely contact, short-distance, and long-distance type as well as the overall abundance of the ectomycorrhizas showed distinct temporal patterns although no specific response to the host seasonal phenology was found. The abundances of the contact type showed no relation to the seasons whereas those of the short- and the long-distance type increased with time. The formation of the long-distance type was strongly influenced by the host species treatments while that of the other two types was not so. Therefore, the different exploration types demonstrate distinct temporal patterns depending on the types but no specific seasonal responses.

Dry bean (Phaseolus vulgaris L.) is an important commercialized field crop in South Africa for aiding in food security as a cheap protein source. However, it is highly susceptible to root-knot nematodes (RKN), Meloidogyne species. Use of indigenous nematophagous fungi as bio-control agents (BCA) of Meloidogyne nematodes is a promising research focus area. This is because indigenous fungal species are naturally part of the ecosystem and therefore compatible with other biological processes unlike most synthetic chemicals. The objective of the study was to identify indigenous nematophagous fungal BCA and establish their potential efficacy in reducing M. enterolobii population densities on dry bean with and without incorporation of compost. Screened indigenous fungal species included Aspergillus terreus, Talaromyces minioluteus, T. sayulitensis, Trichoderma ghanense, and T. viride. There were observed significant parasitism differences (P ≤ 0.05) among the BCA, with T. ghanense showing the highest egg parasitism (86%), followed by T. minioluteus (72%) and T. sayulitensis (70%). On the other hand, the highest J2 parasitism was observed on T. minioluteus (95%), followed by A. terreus and T. viride (63%). A similar trend was observed under in vivo conditions, with higher efficacy with compost incorporation. This provides a highly encouraging alternative and ecologically complementary Meloidogyne management in dry bean production.

This study evaluated the antimicrobial activity, resistance development, and synergistic potential of cell-free supernatant (CFSs) derived from Levilactobacillus brevis (Lb-CFS) and Lactiplantibacillus plantarum (Lp-CFS) against Klebsiella pneumoniae. Both CFSs exhibited potent growth inhibition, with minimum inhibitory concentrations (MICs) of 128 μg/mL and 64 μg/mL for Lb-CFS and Lp-CFS, respectively, and demonstrated dose-dependent bactericidal activity, achieving complete bacterial eradication at minimum bactericidal concentrations (MBC) within 6 h. The CFSs suppressed the expression of virulence genes (galF, wzi, and manC) and biofilm formation in a dose-dependent manner. Synergistic interactions were observed when combining CFSs with antibiotics, resulting in 2- to fourfold reductions in antibiotic MICs and MBCs. Notably, adaptive evolution experiments revealed significantly slower resistance development in K. pneumoniae against CFSs (twofold MIC/MBC increase) compared to antibiotics (16- to 128-fold increase) after 21 days. Furthermore, CFS-adapted strains exhibited increased antibiotic susceptibility, while antibiotic-adapted strains displayed cross-resistance to multiple antibiotics. No cross-resistance occurred between Lb-CFS and Lp-CFS, suggesting distinct adaptive mechanisms. These findings highlight the potential of probiotic-derived CFSs as effective antimicrobials with a lower propensity for inducing rapid resistance compared to conventional antibiotics, suggesting their promise in combating multidrug-resistant infections.

The emergence of Candida auris has caused a major concern in the public health worldwide. This novel fungus is characterized by its multidrug resistance profile, ability to thrive in harsh and stressful conditions, as well as high temperatures and salt concentrations, persistence on hospital surfaces, causing nosocomial infections and outbreaks, and unique fitness properties. Here, we study the antifungal susceptibility patterns, thermotolerance, and halotolerance of 15 putative C. auris clinical isolates from Inkosi Albert Academic Hospital, Durban, South Africa. Five of the C. auris isolates showed resistance to all three antifungals (fluconazole, amphotericin B, and micafungin) and were selected for characterization of their adaptability mechanisms. Four of the tested multidrug-resistant C. auris isolates (C. auris strain F25, C. auris strain F276, C. auris F283, and C. auris M153) showed good growth when exposed to high temperature (42 °C) and salinity (10% NaCl) conditions whereas one isolate (C. auris F65) showed moderate growth under these conditions. Candida parapsilosis showed poor growth whereas C. albicans no growth under these conditions. The five C. auris strains were positive for all the adaptive features.

The spread of microbial resistance is a threat to public health. In this study, the anti-microbial, anti-biofilm, and efflux pump inhibitory effects of ellagic acid-loaded magnetic nanoparticles (Fe₃O₄NPs@EA) against beta-lactamase producing Escherichia coli isolates have been investigated. The effects of Fe₃O₄ NPs@EA on the growth inhibition of E. coli isolates were determined by disc diffusion method and determining the minimum inhibitory concentration was done using broth micro-dilution method. The anti-biofilm effect of nanoparticles was investigated using the microplate method. The efflux pump inhibitory effect of nanoparticles was investigated using cart-wheel method and by investigating the effect of nanoparticles on acrB and tolC genes expression levels. Fe₃O₄ NPs@EA showed anti-bacterial effects against test bacteria, and the MIC of these nanoparticles varied from 0.19 to 1.56 mg/mL. These nanoparticles caused a 43-62% reduction in biofilm formation of test bacteria compared to control. Furthermore, efflux pump inhibitory effect of these nanoparticles was confirmed at a concentration of 1/8 MIC, and the expression of acrB and tolC genes decreased in bacteria treated with 1/4 MIC Fe₃O₄ NPs@EA. According to the results, the use of nanoparticles containing ellagic acid can provide a basis for the development of new treatments against drug-resistant E. coli. This substance may improve the concentration of antibiotics in the bacterial cell and increase their effectiveness by inhibiting the efflux in E. coli isolates.

The intestinal microbiota plays significant role in the physiology and functioning of host organisms. However, there is limited knowledge of the composition and evolution of microbiota-host relationships from wild ancestors to modern domesticated species. In this study, the 16S rRNA gene V3-V4 in the intestinal contents of different pig breeds was analyzed and was compared using high-throughput sequencing. This identified 18 323 amplicon sequence variants, of which the Firmicutes and Actinobacteria phyla and Bifidobacterium and Allobaculum genera were most prevalent in wild pigs (WP). In contrast, Proteobacteria and Firmicutes predominated in Chinese Shanxi Black pigs (CSB), while Firmicutes were the most prevalent phylum in Large White pigs (LW) and Iberian pigs (IB), followed by Bacteroidetes in IB and Proteobacteria in LW. At the genus level, Shigella and Lactobacillus were most prevalent in CSB and LW, while Actinobacillus and Sarcina predominated in IB. Differential gene expression together with phylogenetic and functional analyses indicated significant differences in the relative abundance of microbial taxa between different pig breeds. Although many microbial taxa were common to both wild and domestic pigs, significant diversification was observed in bacterial genes that potentially influence host phenotypic traits. Overall, these findings suggested that both the composition and functions of the microbiota were closely associated with domestication and the evolutionary changes in the host. The members of the microbial communities were vertically transmitted in pigs, with evidence of co-evolution of both the hosts and their intestinal microbial communities. These results enhance our understanding and appreciation of the complex interactions between intestinal microbes and hosts and highlight the importance of applying this knowledge in agricultural and microbiological research.