In vitro kontrola patogenih bakterija poreklom iz humanog materijala delovanjem etarskih ulja i ekstrakata odabranih biljnih vrsta
In vitro control of pathogenic bacteria from human material by essential oils and extracts of certain plant species
Author
Stanković, NemanjaMentor
Mihajilov-Krstev, Tatjana
Committee members
Čomić, Ljiljana
Kocić, Branislav
Zlatković, Bojan
Stankov-Jovanović, Vesna

Metadata
Show full item recordAbstract
Poslednjih decenija jedan od vodećih problema u medicini je stvaranje
rezistentnosti patogenih mikroorganizama na delovanje antibiotika. Bakterije koje
pokazuju značajnu rezistentnost na postojeće antibiotike su: meticilin rezistentni
Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, Acinetobacter spp., itd.
Prilikom delovanja na patogene bakterije, antibiotici neselektivno utiču i na
nepatogene bakterije, izazivajući pri tom nepredvidive genetske promene. Pored
korisnih efekata u delovanju na bakterije, raspoloživi antibiotici, takođe mogu
prouzrokovati neželjene efekte kao što su hipersenzitivnost i imunosupresija. To
je razlog zbog koga se traga za novim agensima sa antibiotskim delovanjem. Jedan od
prirodnih izvora takvih agenasa su etarska ulja i ekstrakti aromatičnih biljaka koji
se koriste u tradicionalnoj medicini za lečenje mnogih infektivnih bolesti i
bolesti koje nastaju kao posledica oksidativnog stresa. Zbog toga je cilj istraživanja
ove disertacije bio da se izvrši ko...mparativna analiza hemijskog sastava,
antibakterijske i antioksidativne aktivnosti odabranih biljnih vrstaAngelica
pancicii, Angelica sylvestris, Laserpitium latifolium, Achillea grandifolia, Achillea
crithmifolia, Artemisia absinthium, Tanacetum parthenium i Hyssopus officinalis koje su
veoma zastupljene u tradicionalnoj medicini.
Materijal i metode. Biljni materijal koji je korišćen u ovom istraživanju je
prikupljan tokom 2012. i 2013. godine na teritoriji jugoistočne Srbije. Nakon
sušenja biljnog materijala, pristupilo se procesu izolovanja etarskih ulja metodom
hidrodestilacije u aparaturi po Klevindžeru, kao i pripremanju metanolnih
ekstrakata alkoholnom ekstrakcijom. Hemijski sastav ulja je analiziran pomoću GH
(gasna hromatografija) i GH/MS (gasna hromatografija sa spektrometrijom masa)
metoda. Ukupni fenoli su određivani metodom po Folin-Sjoklto-u, sa malim
modifikacijama, a ukupni flavonoidi su utvrđivani korišćenjem aluminijum
hlorid (AlCl3) kolorimetrijskog metoda. Antioksidativna aktivnost etarskih ulja i
ekstrakata je određivana pomoću ABTS (2,2'-azino-bis (3-etilbenzotiazolin-6-
sulfonska kiselina), DPPH (2,2-difenil, 1-pikril hidrazil), TRP (total reducing
power) i FRAP (Ferric reducing antioxidant power) metoda. Antibakterijska aktivnost
izolovanih etarskih ulja,metanolnih ekstrakata i četiri antibiotika
(ciprofloksacin, doksiciklin, gentamicin i eritromicin) je ispitivana
mikrodilucionom metodom protiv 16 bakterijskih sojeva poreklom iz briseva rana, grla i nosa, sputuma i aspirata pacijenata (Escherichia coli, Pseudomonas aeruginosa,
Klebsiella sp., Proteus mirabilis, Acinetobacter sp., Staphylococcus aureus, Streptococcus
pyogenes, Streptococcus pneumoniae i Enterococcus faecalis). Sinergističko delovanje
najaktivnijih ulja i antibiotika sa najslabijim delovanjem, eritromicinom, u cilju
snižavanja njegovih aktivnih koncentracija, je određivno uz pomoć metode šahovske
table tj. „checkerboard“ metodom.
During the last several decades one of the leading challenges in medicine
is appearance of resistance of pathogenic microorganisms to antibiotic activity. Bacteria
showing significant resistance on existing antibiotics include: methicillin-resistant
Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, Acinetobacter spp., etc. During
the activity on pathogenic bacteria, the antibiotics simultaneously show non-selective impact
on nonpathogenic bacteria, causing unpredictable genetic changes. In addition to beneficial
effects on bacteria, the available antibiotics may also cause adverse effects such as
hypersensitivity and immunosuppression. Therefore studies of new agents with antibiotic
activity are constantly ongoing. One of the natural sources of such agents are essential oils
and extracts of aromatic plants used in traditional medicine as a cure for many infectious
diseases and ailments caused by oxidative stress. Therefore the goal of study presented in this
dissertation was compar...ative analysis of chemical composition, antibacterial and antioxidant
activity of eight chosen plant species regularly represented in traditional medicine: Angelica
pancicii, Angelica sylvestris, Laserpitium latifolium, Achillea grandifolia, Achillea
crithmifolia, Artemisia absinthium, Tanacetum parthenium and Hyssopus officinalis.
The plant material used in this study was collected in 2012 and 2013
in southeastern Serbia. After the plant material was dried, essential oils were isolated by
hydro-distillation method using the Clevenger apparatus, while methanol extracts were
prepared by alcoholic extraction. The chemical composition of oil was analyzed by GC (gas
chromatography) and GC/MS (gas chromatography with mass spectrometry) methods. The
total amount of phenols was determined by Folin-Ciocalteu method with slight modifications,
while total flavonoids were determined by aluminum chloride (AlCl3) colorimetric method.
The antioxidant activity of essential oils and extracts was determined by ABTS (2,2'-azinobis
(3-ethylbenzothiazoline-6-sulphonic acid), DPPH (2,2-diphenyl-1-picrylhydrazyl), TRP
(total reducing power) and FRAP (Ferric reducing antioxidant power) methods. The
antibacterial activity of isolated essential oils, methanol extracts and four antibiotics
(ciprofloxacin, doxycycline, gentamicin and erythromycin) was studied by micro-dilution
method against 16 bacterial strains collected from swabs of wounds, throat, nose, sputum and
aspirate of patients (Escherichia coli, Pseudomonas aeruginosa, Klebsiella sp., Proteus
mirabilis, Acinetobacter sp., Staphylococcus aureus, Streptococcus pyogenes, Streptococcus
pneumoniae and Enterococcus faecalis). The synergistic activity of most active oils and the
lowest-activity antibiotic (erythromycin), in order to decrease its active concentrations, was determined by the so-called “checkerboard” method.