Journal: Frontiers in Plant Science

Article Title: Development of molecular biomarkers for monitoring of arable crops colonization with Methylobacterium symbioticum SB0023/3, a methylotrophic bacterium commonly used as a biostimulant in agriculture

doi: 10.3389/fpls.2026.1718185

Figure Lengend Snippet: Derivative melting curves for SYBR Green for the xoxF -targeting primer (at 90.3 °C) and for the primers targeting copG and ubik at 84.6 °C and 93.2 °C, respectively. Each peak represents one of the analyzed bacteria. The respective Methylobacterium species are indicated next to the peak. These are: M. symbioticum SB0023/3, M. radiotolerans DSM 760, M. mesophilicum DSM 1708 and M. dankookense SW08-7.

Article Snippet: The reference strains of M. radiotolerans DSM 760, M. mesophilicum DSM 1708 and M. dankookense SW08-7 (DSM 22415) were obtained from DSMZ (German Collection of Microorganisms and Cell Cultures) and cultured on nutrient agar [peptone from casein (10 g/L) (Carl Roth), yeast extract (5 g/L) (Carl Roth) and agar (15 g/L) (Carl Roth)].

Techniques: SYBR Green Assay, Bacteria

Journal: Microbiome

Article Title: Increased caecal Intestinimonas abundance inhibits E. tenella gametogenesis via EtGFAT regulation and alleviates infection through immunity

doi: 10.1186/s40168-025-02302-8

Figure Lengend Snippet: The culture supernatant of I . butyriciproducens inhibits E . tenella sporozoite invasion, with 2′-O-methyladenosine identified as the active inhibitory component. a Flow cytometry was used to evaluate the effects of BC and b CS on E . tenella sporozoite invasion (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). The results indicate that the culture supernatant of I . butyriciproducens significantly inhibited E . tenella sporozoite invasion. The 7-AAD staining method was used to evaluate the direct effects of I . butyriciproducens cells ( c ) and culture supernatant ( d ) on merozoite mortality. These results indicate that neither the bacterial cells nor the culture supernatant exerted a significant lethal effect on the merozoites. Statistical significance was determined by ANOVA. e Nontargeted metabolomics was used to detect the main components in the culture supernatant of I . butyriciproducens . A total of 517 metabolites were identified in positive ion mode, and 314 metabolites were identified in negative ion mode. f Volcano plot of differentially abundant metabolites. In positive ion mode, the abundance of 40 metabolites was significantly increased, whereas the abundance of 11 was significantly decreased; in negative ion mode, the abundance of 30 metabolites was significantly increased, and the abundance of 17 was significantly decreased. g The 70 metabolites with significantly increased abundance from both modes were combined for analysis, and a pie chart was generated to illustrate the proportions of each substance type according to classification. h Flow cytometry was used to assess the effects of six differentially abundant metabolites on sporozoite invasion. The results indicate that only 2′-O-methyladenosine inhibited sporozoite invasion across all the concentration gradients tested. The results of the metabolite cytotoxicity assays are shown in the supplementary figures

Article Snippet: The DF-1 cell line was obtained from the ATCC repository (CRL-3586TM), while I . butyriciproducens was obtained from the DSMZ (DSM 26588, type strain).

Techniques: Flow Cytometry, Staining, Generated, Concentration Assay

Journal: Microbiome

Article Title: Increased caecal Intestinimonas abundance inhibits E. tenella gametogenesis via EtGFAT regulation and alleviates infection through immunity

doi: 10.1186/s40168-025-02302-8

Figure Lengend Snippet: The in vivo anticoccidial effects of I . butyriciproducens cells are significantly greater than those of the I . butyriciproducens cell culture supernatant. a This experiment included six groups: the MOCK, CON, ABX, FMT, I . butyriciproducens bacterial cell groups and the I . butyriciproducens culture supernatant group. AA broilers received ABX treatment starting at 1 day old, and from 10 days old, they were continuously supplemented with I . butyriciproducens bacterial cells or culture supernatant for 5 days before E . tenella infection. b Representative macroscopic images of chicken caeca and c lesion scores from the MOCK, CON, ABX, FMT, I . butyriciproducens cell and I . butyriciproducens culture supernatant groups. The results indicate that I . butyriciproducens cells significantly alleviate caecal lesions induced by E . tenella (** p < 0.01), whereas its culture supernatant has no significant effect. d OPG for each group. I. butyriciproducens significantly reduced E . tenella oocyst shedding (**** p < 0.0001), whereas its culture supernatant had no effect. e Flow cytometry analysis of CD3 + CD8 + T cell proportions in caecal tonsils across groups at 7 dpi, with statistical significance determined by ANOVA ( f ). The results showed that I . butyriciproducens effectively increased CD8 + T cell proportions in the caecal tonsils, while the culture supernatant had no effect

Article Snippet: The DF-1 cell line was obtained from the ATCC repository (CRL-3586TM), while I . butyriciproducens was obtained from the DSMZ (DSM 26588, type strain).

Techniques: In Vivo, Cell Culture, Infection, Flow Cytometry

Journal: Microbiome

Article Title: Increased caecal Intestinimonas abundance inhibits E. tenella gametogenesis via EtGFAT regulation and alleviates infection through immunity

doi: 10.1186/s40168-025-02302-8

Figure Lengend Snippet: I . butyriciproducens confers protection against E . tenella infection by inhibiting the expression of the EtGFAT gene and enhancing T cell-mediated immune responses. a I . butyriciproducens colonisation and sodium butyrate (NaB) supplementation experiments. The NaB group is the sodium butyrate drinking group, which is administered simultaneously with I . butyriciproducens . Sodium butyrate was added to the daily drinking water at a dose of 5 g/L. b Expression of the EtGFAT gene in the CON, ABX, FMT, I . butyriciproducens , and NaB groups was measured by qPCR (** p < 0.01, **** p < 0.0001). c Percentages of Bu-1⁺ cells in peripheral blood across the MOCK, ABX, CON, FMT, I . butyriciproducens and NaB groups at 5, 6 and 7 dpi. The error bars represent the standard deviation (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). d CD3 + CD4 + T cell percentages and e CD3 + CD8 + T cell percentages at 5, 6 and 7 dpi. The bar graphs show the percentage of CD3 + CD4 + T cells in different experimental groups, including the MOCK, CON, ABX, FMT, I . butyriciproducens , and NaB groups, at 5 dpi (left), 6 dpi (middle) and 7 dpi (right). Data are represented as the mean ± standard deviation, with individual data points shown. Secretion of IFN-γ ( f ), IL-17 ( g ) and IL-10 ( h ) at 5, 6 and 7 dpi. The bar graphs show the secretion levels in different experimental groups, including CON, ABX and I . butyriciproducens , at 5 dpi (left), 6 dpi (middle) and 7 dpi (right). Data are presented as the mean ± standard deviation, with individual data points shown (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). i Representative images of caecal tissues from different treatment groups (MOCK, CON, FMT, ABX, I . butyriciproducens , NaB). j Comparison of caecal lesion scores between different treatment groups (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The DF-1 cell line was obtained from the ATCC repository (CRL-3586TM), while I . butyriciproducens was obtained from the DSMZ (DSM 26588, type strain).

Techniques: Infection, Expressing, Standard Deviation, Comparison

Journal: Microbiome

Article Title: Increased caecal Intestinimonas abundance inhibits E. tenella gametogenesis via EtGFAT regulation and alleviates infection through immunity

doi: 10.1186/s40168-025-02302-8

Figure Lengend Snippet: I . butyriciproducens increases caecal short-chain fatty acid levels during the gametogony stage of E . tenella and significantly stimulates CD8⁺ T cells to secrete high levels of IFN-γ in vitro. a Principal component analysis (PCA) plot illustrating sample clustering from different groups (ABX, CON, FMT, IB, and NaB) based on principal components. b Clustered heatmaps illustrating the relative levels of SCFAs across different groups: CON, ABX, FMT, IB and NaB. The colours represent the relative abundance of each SCFA, with red indicating higher levels and blue indicating lower levels. c Flow cytometry analysis of CD4⁻ T cell percentages in CD8⁻ cell complexes (comprising CD4⁺ T cells and other antigen-presenting cells) cocultured with I . butyriciproducens , suggesting that I . butyriciproducens does not significantly stimulate CD4⁺ T cell proliferation. d Flow cytometry analysis of CD8⁺ T cell percentages in CD4⁻ cell complexes cocultured with I . butyriciproducens , indicating that I . butyriciproducens significantly stimulates CD8⁺ T cell proliferation in the absence of CD4⁺ cells or within CD4⁻ cell complexes. e Analysis of CD4⁺ T cell and CD8⁺ T cell percentages (*** p < 0.001). f Analysis of IFN-γ secretion by CD4⁻ cell complexes and CD8⁻ T cell complexes cocultured with I . butyriciproducens (*** p < 0.001)

Article Snippet: The DF-1 cell line was obtained from the ATCC repository (CRL-3586TM), while I . butyriciproducens was obtained from the DSMZ (DSM 26588, type strain).

Techniques: In Vitro, Flow Cytometry