The Impact of Phytochemicals on Antibiotic-Resistant Microbiota and Advanced Biotyping on Multidrug-Resistant Bacteria
Author
Shajarat, Sami
Term
4. term
Publication year
2022
Submitted on
2022-06-01
Pages
35
Abstract
Antibiotic-resistant bacteria are a growing threat to public health. Some bacteria produce enzymes called extended-spectrum beta-lactamases (ESBL) that disable many common antibiotics, leaving fewer treatment options. To help address this problem, our study had two aims: (1) test whether plant-derived compounds from Salicornia ramosissima have antibacterial effects, and (2) evaluate a fast laboratory method that detects the proteins bacteria use to resist antibiotics, with the goal of speeding up antibiotic susceptibility testing. We extracted plant compounds using water and organic solvents and checked for bactericidal (killing bacteria) or bacteriostatic (slowing growth) effects using standard agar diffusion and optical density tests. We also analyzed bacterial proteins with a rapid liquid chromatography–tandem mass spectrometry (LC-MS/MS) workflow, including S-Trap protein preparation and ion-mobility–enabled measurements, to identify resistance mechanisms. At the concentrations available to us, the Salicornia ramosissima extracts showed no measurable bactericidal or bacteriostatic activity. In contrast, the advanced LC-MS/MS approach identified 30 resistance proteins, including three ESBL enzymes (TEM-55, SHV-65, and CTX-M-82). These findings suggest that higher concentrations or different preparations of the plant compounds would be needed to observe antibacterial effects. The LC-MS/MS workflow showed promise for identifying resistance proteins and could support faster, more informative antibiotic susceptibility testing, which merits further evaluation.
Antibiotikaresistente bakterier udgør en voksende trussel mod folkesundheden. Nogle bakterier danner enzymer kaldet udvidet spektrum beta-laktamaser (ESBL), som kan nedbryde mange almindelige antibiotika og dermed begrænse behandlingsmulighederne. For at bidrage til løsninger havde vores studie to mål: (1) teste om plantestoffer fra Salicornia ramosissima har antibakteriel effekt, og (2) vurdere en hurtig laboratoriemetode, der kan påvise de proteiner, bakterier bruger til at modstå antibiotika, med henblik på hurtigere antibiotikafølsomhedstest. Vi ekstraherede plantestoffer med vand og organiske opløsningsmidler og undersøgte for bakteriedræbende (dræber bakterier) eller bakteriehæmmende (hæmmer vækst) effekt ved hjælp af standard agar-diffusion og optisk densitet-målinger. Derudover analyserede vi bakterielle proteiner med en hurtig væskekromatografi–tandem massespektrometri (LC-MS/MS) arbejdsgang, inklusive S-Trap-baseret proteinforberedelse og målinger med ionmobilitet, for at identificere resistensmekanismer. Ved de koncentrationer, vi havde til rådighed, viste ekstrakterne fra Salicornia ramosissima ingen målbar bakteriedræbende eller -hæmmende aktivitet. Til gengæld identificerede den avancerede LC-MS/MS-tilgang 30 resistensproteiner, heraf tre ESBL-enzymer (TEM-55, SHV-65 og CTX-M-82). Resultaterne tyder på, at højere koncentrationer eller andre præparater af plantestofferne er nødvendige for at se en antibakteriel effekt. LC-MS/MS-arbejdsgangen viste lovende muligheder for at påvise resistensproteiner og kan understøtte hurtigere og mere informative antibiotikafølsomhedstest, hvilket bør undersøges nærmere.
[This apstract has been rewritten with the help of AI based on the project's original abstract]