Project description:This study aimed to characterize the non-targeted metabolite profile of Streptomyces sp. LZZY-S40 to explore its potential for producing bioactive secondary metabolites. The strain LZZY-S40 was cultivated on ISP 3 agar plates at 28°C for 7 days. Spores were harvested and inoculated into 250 mL Erlenmeyer flasks containing 50 mL of seed medium (pH 7.2–7.4) for 48 hours at 28°C with shaking at 250 rpm. The seed culture (8% v/v) was then transferred into fermentation medium and incubated under the same conditions for 7 days. After fermentation, the culture broth was centrifuged at 12,000 rpm for 10 minutes to separate the supernatant from the mycelial biomass. The supernatant was extracted three times with ethyl acetate, and the mycelial pellet was extracted with methanol. Organic extracts were concentrated under reduced pressure at temperatures below 40°C. The combined crude extracts were analyzed by untargeted liquid chromatography–mass spectrometry (LC–MS). Data were acquired in both positive and negative ionization modes across an m/z range of 50-1200, with chromatographic separation achieved using a gradient elution program at a flow rate of 400 μL/min and a column temperature of 40°C.
Project description:We analyzed the difference of gene expression between the isolates havested from long-term cultures (Ma et.al. 2020). In this study, the paradaux cell line were cultured for over 40 days under different population control conditions (uncontrolled, negative feedback and paradoxical feedback. Isolates of each culture were harvested at the end of the long-term culture and preped for whole genomic RNA sequencing.
Project description:New and rapid antimicrobial susceptibility/resistance testing methods are required for bacteria from positive blood cultures. In the current study we developed and evaluated a targeted LC-MS/MS assay for the detection of beta-lactam, aminoglycoside and fluoroquinolone resistance mechanisms in blood cultures positive for E. coli or K. pneumoniae. Selected targets were the beta-lactamases SHV, TEM, OXA-1-like, CTX-M-1-like, CMY-2-like, chromosomal E. coli AmpC, OXA-48-like, NDM, VIM and KPC, the aminoglycoside modifying enzymes AAC(3)-Ia, AAC(3)-II, AAC(3)-IV, AAC(3)-VI, AAC(6’)-Ib, ANT(2”)-I and APH(3’)-VI, the 16S-RMTases ArmA, RmtB, RmtC and RmtF, the quinolone resistance mechanisms QnrA, QnrB, AAC(6’)-Ib-cr, the wildtype QRDR of GyrA, and for E. coli, the porins OmpC and OmpF. The developed assay was evaluated using 100 prospectively collected positive blood cultures, 100 negative blood cultures inoculated with isolates that were previously collected from blood cultures, and 48 isolates inoculated with isolates carrying genes of less prevalent resistance mechanisms.