Comparative expression profiling of gliP and cyp51A genes in Aspergillus fumigatus isolates from chickens and poultry feed, Порівняльний аналіз профілів експресії генів gliP та cyp51A в ізолятах Aspergillus fumigatus від курей та зі зразків корму для птиці

<jats:p>Aspergillus fumigatus is an important opportunistic fungal pathogen that frequently contaminates organic substrates on poultry farms, including feed, litter, dust, and airborne particles, thereby posing a significant risk of respiratory infection to birds. The objective of this study was to compare the relative expression levels of two critical target genes, gliP and cyp51A, in poultry- and feed-associated Aspergillus fumigatus isolates, utilizing beta-actin as the internal reference gene. A total of fifteen Aspergillus fumigatus strains were investigated, comprising ten clinical isolates recovered from infected chickens and five control strains obtained from poultry feed samples. Extraction of total RNA and subsequent complementary DNA synthesis were performed, and relative gene expression values were determined using the quantitative real-time polymerase chain reaction (qRT-PCR) via the comparative cycle threshold method. The transcriptional activity of each sample was normalized against beta-actin, and fold-change values were calculated relative to the feed-derived calibrator control group based on the 2–ΔΔCT equation. The experimental data demonstrated a consistent and significant transcriptional activation of both target genes in the chicken-derived group. Specifically, the relative expression levels of the gliP gene, which is directly involved in the nonribosomal peptide synthesis of the immunosuppressive mycotoxin gliotoxin, were highly upregulated in infected chicken isolates compared to the poultry feed-derived control strains, exhibiting mean fold-change values of 5.03±0.29 versus 1.06±0.16, respectively. Concurrently, the expression of the cyp51A gene, which encodes lanosterol 14-alpha-demethylase and serves as a primary target molecule for azole antifungal drugs, was also prominently increased in chicken-derived isolates compared to the controls, with mean fold-change values reaching 4.30±0.39 versus 1.15±0.27, respectively. The presented results suggest that host-associated Aspergillus fumigatus infection in chickens is characterized by the co-activation of pathways governing both mycotoxin production and environmental azole adaptation. Ultimately, the combined expression profile of gliP and cyp51A could serve as a targeted molecular panel for the preliminary screening and surveillance of pathogenicity potential and antifungal resistance characteristics in poultry-associated Aspergillus fumigatus isolates prior to implementing more extensive phenotyping or mutation sequencing assays.</jats:p>