Project description:Oligonucleotide DNA microarrays were used as a platform to compare C. jejuni isolates from feedlot cattle and human clinical cases from Alberta. Comparative genomic hybridization (CGH) analysis was performed on 87 isolates (46 bovine, 41 human) obtained within the same geographical regions and time frame. In addition, We also performed gene association analysis to determine if any genes may be differentially distributed between human and cattle sources or between clusters dominated by either human or cattle isolates (“human enriched” vs “cattle enriched”). Keywords: Comparative Genomic Hybridization; Genomic epidemiology; Gene-association study
Project description:Oligonucleotide DNA microarrays were used as a platform to compare C. jejuni isolates from feedlot cattle and human clinical cases from Alberta. Comparative genomic hybridization (CGH) analysis was performed on 87 isolates (46 bovine, 41 human) obtained within the same geographical regions and time frame. In addition, We also performed gene association analysis to determine if any genes may be differentially distributed between human and cattle sources or between clusters dominated by either human or cattle isolates (âhuman enrichedâ vs âcattle enrichedâ). Keywords: Comparative Genomic Hybridization; Genomic epidemiology; Gene-association study Data from 119 microarrays is included in the dataset, representing the 89 bacterial strains analyzed (87 field isolates, 2 control laboratory strains). Replicate arrays for 20 of the 87 field isolates were included in the dataset, as well as 5 replicates for each of the 2 laboratory controls. An array representing 1546 ORFs from strain NCTC 11168 was used in CGH experiments. CGH was performed by comparing signal from each Tester field isolate analyzed vs. signal from the Control strain NCTC 11168. Values represent mean of triplicate spots.
Project description:Bovine mastitis is the most common disease that affects dairy cattle worldwide and generates millions of losses for cattle breeders. One of the most common pathogens identified in infected milk samples is Staphylococcus aureus. Presently, there is no fast, bacterial species-recognizing test in the diagnostic market.
The aim of this study was bioinformatic detection and characteristic of fibronectin binding protein A (FnBPA) of Staphylococcus aureus (SA) and investigation of this protein in milk samples obtained from cows with diagnosed mastitis.
Bioinformatic detection of potential biomarker for bovine SA obtained analyses of >90.000.000 amino acid sequences. Analyses on FnBPA included, among others: detection of signal peptides, non-classical proteins, antigenicity, and epitopes prediction. To confirm a presence of the fnbA gene in four SA isolates amplification with specific primers was performed. Detection of FnBPA was carried out by immunoblotting. Proteins mixture was separated by SDS-PAGE, and immunoreactivity and selectivity were performed by monoclonal anti-FnBPA antibodies and SA-negative serum.
Bioinformatic analyses showed that FnBPA is a surface, conservative, immunoreactive, and species-specific protein with antigenic potential. Molecular methods confirmed its presence in 100% of isolates, and immunoblotting proved its immunoreactivity and specificity. Thus, it can be considered as potential biomarker in immunodiagnostic of mastitis.
Project description:Investigation of baseline transcription activity of two different clinical isolates of Staphylococcus aureus with two different susceptibility levels to the antibiotics Vancomycin and Daptomycin.
Project description:A comparative genomic approach was used to identify large sequence polymorphisms among Mycobacterium avium isolates obtained from a variety of host species. DNA microarrays were used as a platform for comparing mycobacteria field isolates with the sequenced bovine isolate Mycobacterium avium subsp. paratuberculosis (Map) K10. ORFs were classified as present or divergent based on the relative fluorescent intensities of the experimental samples compared to Map K10 DNA. Map isolates cultured from cattle, bison, sheep, goat, avian, and human sources were hybridized to the Map microarray. Three large deletions were observed in the genomes of four Map isolates obtained from sheep and four clusters of ORFs homologous to sequences in the Mycobacterium avium subsp. avium (Maa) 104 genome were identified as being present in these isolates. One of these clusters encodes glycopeptidolipid biosynthesis enzymes. One of the Map sheep isolates had a genome profile similar to a group of Mycobacterium avium subsp. silvaticum (Mas) isolates which included four independent laboratory stocks of the organism traditionally identified as Maa strain 18. Genome diversity in Map appears to be mostly restricted to large sequence polymorphisms that are often associated with mobile genetic elements. Keywords: Comparative genomic hybridization
Project description:Investigation of baseline transcription activity of two different clinical isolates of Staphylococcus aureus with two different susceptibility levels to the antibiotics Vancomycin and Daptomycin. Two different strains of Staphylococcus aureus, one that is fully Vancomycin and Daptomycin Sensitive and one with decreased Vancomycin and Daptomycin Sensitivity - grown to mid-log phase in rich broth.
Project description:To explore the difference in the expression profile of Staphylococcus aureus in chronic periprosthetic joint infection (C_PJI) and acute PJI (A_PJI), we assayed S. aureus isolates from 4 C_PJI and 4 A_PJI by RNA-sequencing.
Project description:Hybridisation of reference strains to the VirEp Staphylococcus aureus microarray, and characterisation of different S. aureus isolates from different locations and associated with different diseases.
