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Vpliv polžje sline na rast in tvorbo biofilma pri bakterijah Staphylococcus aureus in Pseudomonas aeruginosa
Žlender, Tanja (Author), Starčič Erjavec, Marjanca (Mentor) More about this mentor... This link opens in a new window, Tkalec, Valerija (Co-mentor)

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Abstract
Polžji slini pripisujejo protimikrobno in protivnetno delovanje ter vzpodbujanje celične regeneracije. Namen magistrske naloge je bil raziskati učinek polžje sline hrapavega vrtnega polža (Helix aspersa Müller) na rast celic in tvorbo biofilma pri bakterijah Staphylococcus aureus in Pseudomonas aeruginosa v optimiziranem sistemu za določanje biofilma. Bakteriji S. aureus in P. aeruginosa sta pogosto izolirani iz kroničnih ran. Gre za oportunistična patogena, ki pri ljudeh povzročata širok spekter lokalnih in sistemskih okužb. Njuno zdravljenje zaradi pojavljanja odpornosti proti številnim uveljavljenim protimikrobnim zdravilom postaja vse večji izziv. Zato so raziskave novih pristopov za obvladovanje okužb zelo aktualne. V nalogi smo najprej optimizirali sisteme za testiranje vpliva polžje sline na rast celic in tvorbo biofilma. V sklopu tega smo testirali dve obliki polistirenskih mikrotitrskih ploščic (z ravnim in ukrivljenim dnom), gojišča LB, BHI in TSB brez dodane glukoze in z dvema različnima koncentracijama glukoze (0,2 in 2 %), določili protokol za pripravo testnih bakterijskih kultur (gojenje bakterijske kulture s stresanjem in brez ter čas inkubacije bakterijske kulture pred testiranjem polžje sline) ter optimizirali protokol za merjenje biofilma. Izmed sedmih sevov bakterije S. aureus in šestih sevov bakterije P. aeruginosa, pridobljenih iz kroničnih ran, smo izbrali po dva najboljša tvorca biofilma, ki smo ju uporabili v poskusih s polžjo slino. Za testiranje vpliva polžje sline na rast celic in tvorbo biofilma smo izbrali optimalne parametre sistema: mikrotitrsko ploščico z ukrivljenim dnom, gojišče BHI brez dodane glukoze za bakterijo P. aeruginsa in gojišče BHI z 2 % glukozo za bakterijo S. aureus ter 10-urno bakterijsko kulturo v eksponentni fazi rasti. Ugotovili smo, da filtrat naravne polžje sline ne zavira rasti celic in tvorbe biofilma oz. ju v nekaterih primerih, sploh pri bakteriji P. aeruginosa, lahko tudi vzpodbuja.

Language:Slovenian
Keywords:bakterijski biofilmi, tvorba biofilma, Staphylococcus aureus, Pseudomonas aeruginosa, vrtni polž, Helix aspersa Müller, polžja slina, protimikrobne snovi, naravne učinkovine
Work type:Master's thesis/paper (mb22)
Tipology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Year:2020
Publisher:[T. Žlender]
UDC:579.6:547.9:636.95:594.3:616-008.843.1
COBISS.SI-ID:35821315 This link opens in a new window
Views:229
Downloads:90
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Secondary language

Language:English
Title:The effect of snail mucus on growth and biofilm production of Staphylococcus aureus and Pseudomonas aeruginosa
Abstract:
Snail mucus is suggested to have the ability to act as an antimicrobial and anti-inflammatory agent as well as to stimulate cell regeneration. The purpose of this master thesis was to investigate the effect of snail mucus collected from garden snails (Helix aspersa Müller) on cell growth and biofilm formation of bacteria Staphylococcus aureus and Pseudomonas aeruginosa in an optimized testing system. S. aureus and P. aeruginosa are opportunistic pathogens often isolated from chronic wounds. They can cause a wide range of local and systematic infections in humans. Their treatment is becoming increasingly challenging due to the emergence of drug resistance. Therefore, many studies are focused on finding new approaches of infection control. In our thesis, we first optimized a system for testing the effect of snail mucus on cell growth and biofilm formation. As part of the optimization, we tested two types of polystyrene 96-well plates (flat and curved bottom), three different media (LB, BHI, TSB) without added glucose and with two different glucose concentrations (0.2 and 2 %) and optimized preparation of testing culture (with/without shaking during growth and time of culture growth prior testing). We tested the effects of snail mucus on two out of seven S. aureus strains and two out of six P. aeruginosa strains isolated from chronic wounds, which produced the highest amounts of biofilm. The testing was conducted on non-shaked 10-hour bacterial cultures in the exponential growth phase using following system parameters: curved bottom 96-well plate, BHI medium for P. aeruginosa and BHI with 2 % glucose for S. aureus. We found that natural snail mucus filtrate does not inhibit any cell growth nor any biofilm formation. On contrary, it can stimulate growth and biofilm formation in some cases, especially in P. aeruginosa.

Keywords:bacterial biofilms, biofilm formation, Staphylococcus aureus, Pseudomonas aeruginosa, garden snail, Helix aspersa Müller, snail mucus, antimicrobial substances, natural substances

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