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Delovanje izvlečkov rastline Peucedanum ostruthium na bakterije vrste Campylobacter jejuni
Jurhar, Andreja (Author), Smole Možina, Sonja (Mentor) More about this mentor... This link opens in a new window

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Abstract
Bakterije vrste Campylobacter povzročajo kampilobakteriozo, ki v Evropi velja za najpogostejšo bakterijsko črevesno okužbo ljudi. Zaradi prilagoditve C. jejuni na stresne razmere in odpornosti proti enemu ali več antibiotikov so potrebne raziskave o novih protimikrobnih učinkovinah ali takih, ki bi obstoječim antibiotikom povrnile delovanje. Te učinkovine lahko najdemo tudi v naravi. V tej nalogi smo raziskovali protimikrobno in odpornostno-modulatorno delovanje etanolnega in heksanskega izvlečka ter eteričnega olja korenike jaščarice (Peucedanum ostruthium) ter frakcij teh pripravkov. Protimikrobno in odpornostno-modulatorno delovanje smo ovrednotili z minimalno inhibitorno koncentracijo (MIK) z metodo mikrodilucije v bujonu. Izvlečki so na referenčnem sevu C. jejuni z vrednostjo MIK 500 mg/L in več pokazali zmerno protimikrobno delovanje, z izjemo nekaterih frakcij z boljšim učinkom (do 125 mg/L). Test smo opravili tudi na mutantih z izbrisom v genih za izlivne črpalke. Ključno vlogo pri izčrpavanju protimikrobnih snovi iz bakterije C. jejuni ima izlivna črpalka CmeABC. S primerjavo vrednosti MIK antibiotikov in antibiotikov v kombinacijah z izvlečki smo ugotovili, da imajo izvlečki šibko odpornostno-modulatorno delovanje. Ker nas je zanimal mehanizem delovanja izvlečkov P. ostruthium, smo opravili test kopičenja etidijevega bromida ter test membranske integritete. S testiranjem kopičenja etidijevega bromida v štirih različnih koncentracijah (1/4 MIK, 1/8 MIK, 1/16 MIK in 1/32 MIK) smo dokazali, da izvlečki delujejo kot inhibitorji izlivnih črpalk, kar smo zaznali kot povečanje fluorescenčnega signala v celici. Z metodo membranske integritete smo dokazali, da izvlečki delujejo tudi na povečanje prepustnosti celične membrane.

Language:Slovenian
Keywords:Campylobacter jejuni, jaščarica, Peucedanum ostruthium, rastlinski izvlečki, protimikrobne snovi, odpornost na protimikrobna sredstva, izlivne črpalke, membranska integriteta
Work type:Master's thesis/paper (mb22)
Tipology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Year:2018
Publisher:[A. Jurhar]
UDC:579.24/.26:547.9:615.33
COBISS.SI-ID:4955768 This link opens in a new window
Views:835
Downloads:479
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Secondary language

Language:English
Title:Activity of Peucedanum ostruthium extracts on Campylobacter jejuni
Abstract:
Campylobacteriosis is a zoonosis caused by Campylobacter spp. and it is one of the most common intestinal infections of humans in Europe. Due to adaptation of C. jejuni to stress conditions and antimicrobial resistance to one or more antibiotics, research for new antibiotics and active substances that can revive the function of existing antibiotics is needed. These substances can also be found in nature; in our case in extracts and their fractions of the plant Peucedanum ostruthium. We evaluated the antimicrobial and modulatory effectiveness on P. ostruthium with minimal inhibitory concentration (MIC) by the microdilution method in broth. Extracts exhibited from medium to weak antimicrobial activity as the minimal inhibitory concentrations were 500 mg/L or more. The exceptions were two fractions of ethanol extract; E0 (250 mg/L) and E4 (125 mg/L). We have proven that CmeABC pump is the target of P. ostruthium extracts by testing antimicrobial activity on mutants with interrupted gene frame shifts of efflux pumps. By comparing MICs of antibiotics and antibiotics in combinations with three basic extracts we proved week modulatory effects. By testing the accumulation of ethidium bromide in four different concentrations (1/4 MIC, 1/8 MIC, 1/16 MIC and 1/32 MIC) in the C. jejuni cells we found that P. ostruthium extracts function as inhibitors of efflux pumps. By testing membrane integrity we proved that the extracts of P. ostruthium increase the permeability of the C. jejuni cell membrane.

Keywords:Campylobacter jejuni, masterwort, Peucedanum ostruthium, plant extracts, antimicrobials, antimicrobial resistance, efflux pumps, membrane integrity

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