Project description:Purpose:To uncover the related mechanisms underlie virulence attenuation of Brucella canis MucR mutant strain. Methods:Three Brucella canis RM6/66 strains and three Brucella canis ΔmucR strains were grown in TSB at 37℃ until the log phase was reached, total RNA was isolated using the TRIzol according to the manufacturer’s instructions.The sequencing library of each RNA sample was prepared by using NEB Next Ultra Directional RNA Library Prep Kit for Illumina as recommended by the manufacturer. An Illumina platform was used to perform the transcriptome sequencing. Results: The results revealed that expressions of 694 genes were significantly different between RM6/66 and ΔmucR. Data analysis showed that in the COG term, the different expressed genes involved in translation, ribosomal structure and biogenesis, signal transduction mechanisms, energy production and conversion, intracellular trafficking, secretion, and vesicular transport, and extracellular structures were significantly affected. Pathway enrichment analysis indicated that the genes involved in ribosome, oxidative phosphorylation, aminoacyl-tRNA biosynthesis and protein export were significantly enriched.
Project description:Canine babesiosis, a tick-borne disease, is characterized by protozoan parasites invading red blood cells. It is rapidly expanding in many European countries. Examining extracellular vesicles (EVs) and their protein cargo has the potential to offer crucial insights into the response to Babesia canis infection, presenting opportunities for advancements in veterinary diagnostic and therapeutic strategies. In the present study, we have a) isolated small EVs (< 200 nm) from the serum of 15 healthy dogs and 15 dogs naturally infected with B. canis using size-exclusion chromatography (fraction 2 and 3 per each sample), (2) characterized isolated EVs by nanoparticle tracking analysis, transmission electron microscopy and Western blot (3) analysed the protein cargo of isolated EVs by mass spectrometry. We hypothesized that there will be a difference in EV characteristics (size, concentration, EV marker proteins) and profiles of luminal proteins between the two experimental groups. Our aim was to characterize proteins that can offer valuable insights into B. canis infection in dogs, thereby unravelling the complex mechanisms of B. canis infection.
Project description:Streptococcus agalactia were isolated from virginal swabs and from bovine. the experiment aims at differentiating the proteome and metabolome differences between the 2 isolated strains. To achieve this target, label free proteomics and untargeted metabolomics profiling were applied
Project description:Streptococcus pneumoniae parental T4R and Δ1434-8 strains were cultured in quadruplicated in C+Y medium. Proteins were isolated, quantified, trypsin digested, and analyzed by 1D LC ESI MS/MS.
Project description:The microbiota of the mouth disperses into the lungs and both compartments share similar phyla. Considering the importance of the microbiota in the maturation of the immunity and physiology during the first days of life, we hypothesized that primo-colonizing bacteria of the oral cavity may induce immune responses in bronchial epithelial cells. Herein, we have isolated and characterized 57 strains of the buccal cavity of two human new-borns. These strains belong to Streptococcus, Staphylococcus, Enterococcus, Rothia and Pantoea genera; Streptococcus being the most represented. The strains were co-incubated with a bronchial epithelial cell line (BEAS-2B) and we established their impact on a panel of cytokines/chemokines and global changes in gene expression. The Staphylococcus strains, which appeared soon after birth, induced a high production of IL-8, suggesting they can trigger inflammation, whereas the Streptococcus strains were less associated with inflammation pathways. The genera Streptococcus, Enterococcus and Pantoea induced differential profiles of cytokine/chemokine/growth factor and set of genes associated with maturation of morphology. Altogether, our results demonstrate that the micro-organisms, primo-colonizing the oral cavity, impact immunity and morphology of the lung epithelial cells, with specific effects depending on the phylogeny of the strains.
