M. Sc. Thesis
Povzetek
The emergence of multidrug-resistant (MDR) bacterial strains represents a major health concern that requires research into alternative methods for managing bacterial infections. Our aim was to test the effect of a new potential agent - human amniotic membrane (hAM) homogenate - on biofilm formation of selected antibiotic-resistant uropathogenic Escherichia coli (UPEC) strains, including MDR strains. In addition, we screened the UPEC strains for the presence of virulence-associated genes. Ten different UPEC strains were employed and tested for their ability to form biofilms. We then selected four of these UPEC strains (DL 9, DL 13, DL 14 and DL 88) for further investigation of the effect of the hAM homogenate. The hAM homogenates were prepared in phosphate-buffered saline (PBS) or ADMEM + F12 medium. Cultures grown overnight in Luria-Bretani (LB) were diluted in fresh LB, PBS or hAM homogenate, inoculated into a 96-well microtiter plate and incubated at 37°C for 24 hours. We aimed to evaluate the effect of the hAM homogenate on the biofilm-forming ability of the UPEC strains using crystal violet assay and colony-forming units (CFU) determination assay. Three of the four UPEC strains tested (DL9, DL13 and DL88) were found to have the highest capacity for biofilm formation in LB, an intermediate capacity in PBS and the lowest capacity for biofilm formation in the hAM homogenate. The hAM homogenate was found to negatively affect the biofilm-forming ability of these UPEC strains, indicating its potential as an alternative treatment strategy against UPEC.
Ključne besede
Escherichia coli;UPEC;urinary tract infections;biofilm;human amniotic membrane;human amniotic membrane homogenate;antimicrobial peptides;
Podatki
Jezik: |
Angleški jezik |
Leto izida: |
2024 |
Tipologija: |
2.09 - Magistrsko delo |
Organizacija: |
UL MF - Medicinska fakulteta |
Založnik: |
[M. Najdovska] |
UDK: |
579.2:616.6 |
COBISS: |
207678467
|
Št. ogledov: |
40 |
Št. prenosov: |
1621 |
Ocena: |
0 (0 glasov) |
Metapodatki: |
|
Ostali podatki
Sekundarni jezik: |
Slovenski jezik |
Sekundarni naslov: |
Učinek homogenata humane amnijske membrane na uropatogeno bakterijo Escherichia coli in njeno sposobnost tvorbe biofilma |
Sekundarni povzetek: |
Pojav večkratno odpornih bakterijskih sevov predstavlja velik problem javnega zdravja in je zato nujno potrebno raziskovati alternativne metode za obvladovanje bakterijskih okužb. Naš cilj je bil preizkusiti učinek novega potencialnega sredstva - homogenata humane amnijske membrane (hAM) - na tvorbo biofilma izbranih proti antibiotiokom odpornih uropatogenih Escherichia coli (UPEC) sevov, vključno z večkratno odpornimi sevi. Poleg tega smo testirali seve UPEC na prisotnost genov, povezanih z virulenco. Uporabljenih je bilo deset različnih sevov UPEC, pri katerih smo testirali sposobnost tvorbe biofilmov. Glede na rezultate smo nato izbrali štiri seve UPEC (DL 9, DL 13, DL 14 in DL 88) za nadaljnjo preiskavo učinka homogenata hAM. Homogenate smo pripravili v fosfatnem pufru (PBS) ali ADMEM + F12 mediju. Kulture, ki so zrasle preko noči v tekočem gojišču Luria-Bretani (LB), smo redčili v svežem LB, PBS ali homogenatu hAM, inokulirali v mikrotitrsko ploščo s 96 vdolbinicami in 24 ur inkubirali pri 37 °C. Z barvanjem mase biofilma s kristalno vijoličnim in določanjem število celic v biofilmu smo ovrednotili učinek homogenata hAM na sposobnost sevov UPEC za tvorbo biofilma. Ugotovili smo, da imajo 3 od 4 testiranih sevov UPEC (DL9, DL13 in DL88) največjo zmogljivost za tvorbo biofilma v LB, srednjo zmogljivost v PBS in najnižjo zmogljivost za tvorbo biofilma v homogenatu hAM. Ugotovili smo, da homogenat hAM negativno vpliva na sposobnost tvorbe biofilmov pri teh sevih UPEC in ima zato lahko potencial kot alternativno terapevtsko sredstvo proti UPEC. |
Sekundarne ključne besede: |
Escherichia coli;UPEC;okužbe urinarnega trakta;humana amnijska membrana;biofilm;homogenat humane amnijske membrane;protimikrobni peptidi; |
Vrsta dela (COBISS): |
Magistrsko delo/naloga |
Študijski program: |
0 |
Komentar na gradivo: |
Univ. v Ljubljani, Biotehniška fak., Oddelek za mikrobiologijo |
Strani: |
1 spletni vir (1 datoteka PDF (XI, 55 str.)) |
ID: |
25050722 |