Archives of Microbiology最新文献

Panax ginseng is a precious Chinese medicinal plant with a long growth cycle and high medicinal value. Therefore, it is of great significance to explore effective ways to increase its yield and main active substance content to reduce the cost of ginseng, which is widely used in food and clinical applications. Here, we review the key roles of microorganisms in the biological control of ginseng diseases, enhancement of ginseng yield, biotransformation of ginsenosides, and augmentation of ginsenoside bioactivity. The application of microorganisms in P. ginseng faces multiple challenges, including the need for further exploration of efficient microbial strain resources used in the cultivation of ginseng and biotransformation of ginsenosides, lack of microbial application in large-scale field cultivation of ginseng, and unclear mechanism of microbial transformation of ginsenosides. This review provides a deeper understanding of the applications of microorganisms in P. ginseng.

Three new strains of Phaffia rhodozyma yeast have recently been isolated in Poland. The aim of this study was to phenotypically characterize these strains and to compare them with the properties of the reference strain. The potential for carotenoid biosynthesis in these strains was also determined, depending on temperature, carbon, and nitrogen sources in the medium. Phaffia rhodozyma yeasts were also identified by MALDI-TOF MS. There were minor differences in cell morphology among the strains. All strains reproduced asexually by budding and formed spherical chlamydospores. No ability for sexual reproduction was observed. Physiological tests showed minor variations between the reference strain and the isolates, likely due to the geographical specificity of the habitat from which they were originally isolated. Analysis of protein spectra showed that the tested yeast isolates had seven common peaks of different intensities, with masses at 2200, 2369, 3213, 3628, 3776, 3921, and 4710 m/z. Moreover, additional strain-dependent spectra were found. The amount of synthesized carotenoids varied with the carbon and nitrogen sources used, as well as the temperature. The best producer of carotenoids was the P. rhodozyma CMIFS 102 isolate.

Bacterial cells often exist in the form of sessile aggregates known as biofilms, which are polymicrobial in nature and can produce slimy Extracellular Polymeric Substances (EPS). EPS is often referred to as a biofilm matrix and is a heterogeneous mixture of various biomolecules such as polysaccharides, proteins, and extracellular DNA/RNA (eDNA/RNA). In addition, bacteriophage (phage) was also found to be an integral component of the matrix and can serve as a protective barrier. In recent years, the roles of proteins, polysaccharides, and phages in the virulence of biofilms have been well studied. However, a mechanistic understanding of the release of such biomolecules and their interactions with antimicrobials requires a thorough review. Therefore, this article critically reviews the various mechanisms of release of matrix polymers. In addition, this article also provides a contemporary understanding of interactions between various biomolecules to protect biofilms against antimicrobials. In summary, this article will provide a thorough understanding of the functions of various biofilm matrix molecules.

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Staphylococcus aureus-induced mastitis is a serious disease in dairy bovine, with no currently effective treatment. Antibiotics demonstrate certain therapeutic potency in dairy husbandry; they generate drug-resistant bacteria, thereby harming public health. LncRNAs and m⁶A have been verified as potential targets in infectious diseases and have powerful regulatory capabilities. However, the biological regulation of lncRNAs with m⁶A modification in mastitis needs further investigation. This study aims to determine the m⁶A-modified lncRNAs in bovine mammary epithelial cells and their diversity during S. aureus induction. Heat-inactivated S. aureus was used to develop the cell injury model, and we subsequently found low cell viability and different m⁶A modification levels. Our analysis of m⁶A-modified lncRNA profiles through MeRIP-seq revealed significant differences in 140 peaks within 130 lncRNAs when cells were injured by S. aureus. Furthermore, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed that these differential m⁶A-modified lncRNAs were mainly enriched in the WNT pathway, and their functions were associated with amino acid metabolism, lipid translocation, and metalloproteinase activity. Here, we report for the first time lncRNAs with m⁶A modification in regulating S. aureus infection, revealing potential mechanisms and targets of infectious diseases, such as mastitis, from an epigenetics perspective.

Biofilms are structured microbial communities encased in a matrix of self-produced extracellular polymeric substance (EPS) and pose significant challenges in various industrial cooling systems. A nuclear power plant uses a biocide active-bromide for control of biological growth in its condenser cooling system. This study is aimed at evaluating the anti-bacterial and anti-biofilm efficacy of active-bromide against planktonic and biofilm-forming bacteria that are commonly encountered in seawater cooling systems. The results demonstrated that active-bromide at the concentration used at the power plant (1 ppm) exhibited minimal killing activity against Pseudomonas aeruginosa planktonic cells. The bacterial cell surface hydrophobicity assay using Staphylococcus aureus and P. aeruginosa indicated that Triton-X 100 significantly decreased the hydrophobicity of planktonic cells, enhancing the susceptibility of the cells to active-bromide. Biofilm inhibition assays revealed limited efficacy of active-bromide at 1 ppm concentration, but significant inhibition at 5 ppm and 10 ppm. However, the addition of a surfactant, Triton-X 100, in combination with 1 ppm active-bromide displayed a synergistic effect, leading to significant biofilm dispersal of pre-formed P. aeruginosa biofilms. This observation was substantiated by epifluorescence microscopy using a live/dead bacterial assay that showed the combination treatment resulted in extensive cell death within the biofilm, as indicated by a marked increase in red fluorescence, compared to treatments with either agent alone. These findings suggest that active bromide alone may be insufficient for microfouling control in the seawater-based condenser cooling system of the power plant. Including a biocompatible surfactant that disrupts established biofilms (microfouling) can significantly improve the efficacy of active bromide treatment.

Polyunsaturated fatty acids (PUFA) are vital molecules in the pharmaceutical, medical, and nutritional industries. Exploration of bacterial strains capable of producing significant amounts of PUFAs offers a promising avenue for biotechnological applications and industrial-scale production. However, an extensive screening of several samples from diverse sources is highly needed to identify a potential strain. The present study provides the results of the evaluation of 15 different screening methodologies (including changes in existing protocols in terms of reagent concentration, incubation temperature and time) for identifying PUFA-producing bacteria in comparison to the gold standard method (Gas chromatography-mass spectrometry), for the first time. The results determined the most effective techniques for each critical PUFA, leading to an optimized screening process that saves time and resources. The H₂O₂ plate assay using 0.5% or 1% H₂O₂ for 72 & 96 h of incubation at 15 °C consistently outperformed others for finding bacteria containing total nutritionally important long chain-PUFA (LC-PUFA), linoleic acid, and arachidonic acid. Whereas the 2,3,5-triphenyl tetrazolium chloride broth assay at 10–15 °C was the most effective and semiquantitative screening methodology for eicosapentaenoic acid (EPA) and alpha-linolenic acid-containing bacteria. Apart from the methodological perspectives, the study also revealed certain potential strains to be targeted in the ongoing research on PUFA-containing bacteria. Further, the manuscript forms the first report on the presence of docosahexaenoic acid (DHA) in Shewanella decolorationis, EPA in Psychrobacter maritimus and Micrococcus aloeverae, and both EPA and DHA in Arthrobacter rhombi. Altogether, the paper generates several thought-provoking insights on the methodological perspectives and identifies potential PUFA-containing bacteria with practical applications in future bacteria-based PUFA research.

Strains M29^T and ASUV-10-1^T, which are aerobic, non-flagellated, and Gram-stain-negative, were isolated from soil samples collected in Inje (37°57’49.1"N 128°19’53.7"E) and Cheonan City (36°48’47.1"N 127°05’22.4"E), South Korea. Phylogenetic analyses based on rRNA gene sequences revealed that strains M29^T and ASUV-10-1^T form a distinct branch within the family Hymenobacter (order Cytophagales, class Cytophagia). Strain M29^T is most closely related to Hymenobacter rubidus DG7B^T with a 16 S rRNA gene sequence similarity of 97.05%. Strain ASUV-10-1^T shows closest genetic similarity to Hymenobacter frigidus B1789^T (96.42%), Hymenobacter jeongseonensis BT683^T (95.97%), and Hymenobacter terricola 3F2T^T (95.65%). The optimal growth conditions for these strains are pH 7.0, no NaCl, and a temperature of 25 °C. The dominant cellular fatty acids identified in these strains are iso-C_15:0, anteiso-C_15:0, and Summed Feature 3 (C_16:1ω 7c / C_16:1ω 6c). Both strains predominantly contain MK-7 as the respiratory quinone. The major polar lipids in strains M29^T and ASUV-10-1^T are phosphatidylethanolamine, aminophospholipid, and aminolipid. Based on biochemical, chemotaxonomic, and phylogenetic data, it is evident that M29^T and ASUV-10-1^T represent new species within the genus Hymenobacter. The new species were classified based on biochemical and chemotaxonomic characteristics. The taxonomic classification of these species was conducted following the guidelines and protocols outlined in Bergey’s Manual of Systematic Bacteriology. We followed the methods for determining physiological and biochemical characteristics, as well as chemotaxonomic markers such as fatty acid profiles, quinone types, and polar lipid compositions. We also compared with the results of carbohydrate utilization and enzyme activities results [Bergey 1994]. Therefore, we propose the names Hymenobacter mellowenesis for strain M29^T (= KCTC 102056^T = NBRC 116578^T) and Hymenobacter aranciens for strain ASUV-10-1^T (= KCTC 92969^T = NBRC 116575^T).

Melatonin (MLT) is a methoxyindole that has potent antioxidant actions, anti-inflammatory, and antiapoptotic capacity. However, its in vitro antibacterial capacity has been the least studied of its properties. Dimethylsulfoxide (DMSO) has been the most used solvent for these tests, but it shows an antimicrobial effect if it is not dissolved. Cyrene™ is a new solvent that has emerged as an alternative to DMSO. Therefore, this study aimed to determine the antimicrobial capacity of MLT by MIC assays, using Cyrene™ as a solvent. Likewise, the solubility of MLT in this solvent and whether it exerted any effect on bacterial growth at different percentages was also determined. Different dilutions of MLT in Cyrene™ with different concentrations, were prepared. No growth inhibition caused by MLT was observed. The growth inhibition observed was because of Cyrene™. The maximum amount of MLT that can be diluted in 100% Cyrene is 10 mg/mL, but this percentage of solvent shows a bactericidal effect. Therefore, it must be dissolved at 5% to avoid this effect, so only 4 mg/mL of MLT can be diluted in it. Therefore, if no other solvents are available, the in vitro antibacterial role of MLT cannot be adequately assessed.

Bacterial biofilms pose significant challenges due to their association with antibiotic resistance, metabolic adaptation, and survival under harsh conditions. Among notable pathogens forming biofilms, Staphylococcus aureus and Acinetobacter baumannii are concerning pathogens in nosocomial settings. However, their behaviour under acidic (pH 4.5) and alkaline (pH10.5) conditions, especially in co-culture setups, remains insufficiently understood. This study investigates these aspects, by examining growth rates, biofilm formation, pH shifts, phenotypic analysis, and gene expression profiles. The results showed A. baumannii exhibited reduced  growth and biofilm formation at pH 4.5, while S. aureus showed slow growth and low biofilm formation at pH10.5 in mono-cultures. S. aureus leaned towards an acidic pH (6–6.5), whereas A. baumannii shifted towards an alkaline pH (8–9). In co-culture environments, growth rates and biofilm formation increased across all pH conditions, converging towards a neutral pH over time. Phenotypic motility assays indicated that A. baumannii exhibited greater motility in alkaline conditions, while S. aureus showed increased staphyloxanthin production under acidic conditions. Gene expression analyses revealed that the fibronectin-binding protein A (FnbA) and N-acetylglucosaminyl-transferase (icaA) genes, responsible for initial attachment during biofilm formation, were highly expressed in acidic co-culture condition but poorly expressed in alkaline condition. In A. baumannii, the outer membrane protein A (OmpA) gene associated with adhesion and virulence, was upregulated in co-culture. The LuxR gene involved in quorum sensing was upregulated in acidic conditions and poorly expressed at pH 10.5. This study elucidates the metabolic adaptability and biofilm formation tendencies of S. aureus towards acidic conditions and A. baumannii towards alkaline conditions, providing insights for better management of biofilm-related infections.

In the fields of cultured meat, biopharmaceuticals, cell therapy, and tissue engineering, large numbers of mammalian cells are required; thus, highly-concentrated cell cultures are widely adopted. In general, such cultures can lead to cell damage caused by waste product accumulation and nutritional inadequacy. In this study, a novel co-culture system where the recombinant lactate-assimilating cyanobacterial strain, KC0110, derived from euryhaline Picosynechococcus sp. PCC 7002, and mammalian muscle cells cultured across porous membranes been developed. By using the KC0110 strain, the amount of ammonium and lactate excreted from C2C12 mouse muscle cells into the culture significantly decreased. Importantly, pyruvate and some amino acids, including pyruvate-derived amino acids, also increased significantly compared to those in monoculture of C2C12 cells. It is believed that the organic acids secreted by the KC0110 strain enhance the growth of mammalian cells, leading to a reduction in high-concentration culture-induced mammalian cell damage [lactate dehydrogenase (LDH) release] through cyanobacterial co-culture. These results show that, through co-cultivation with cyanobacteria, it is possible to culture mammalian cells, alleviating cell damage, even in highly-concentrated cultures. This study demonstrated an in vitro "symbiotic circular system" that can interchange metabolites produced by phototrophs and mammalian cells.