Vpliv mutacij v genih, pomembnih za tvorbo biofilma na adhezijo in invazijo bakterij campylobacter jejuni v celicah caco-2 : magistrsko delo

Damijan, Tjaša

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Title:	Vpliv mutacij v genih, pomembnih za tvorbo biofilma na adhezijo in invazijo bakterij campylobacter jejuni v celicah caco-2 : magistrsko delo
Authors:	ID Damijan, Tjaša (Author) ID Langerholc, Tomaž (Mentor) More about this mentor... ID Šikić Pogačar, Maja (Comentor) ID Klančnik, Anja (Comentor)
Files:	MAG_Damijan_Tjasa_2022.pdf (2,08 MB) MD5: B62B4B44D82C3E023C202F4C66EB5A72
Language:	Slovenian
Work type:	Master's thesis/paper
Typology:	2.09 - Master's Thesis
Organization:	FKBV - Faculty of Agriculture and Life Sciences
Abstract:	Mikroaerofilne bakterije Campylobacter jejuni so kljub zahtevnim pogojem rasti zelo razširjeni mikroorganizmi in najpogostejši povzročitelji akutnega bakterijskega gastroenteritisa pri ljudeh po vsem svetu. Predpostavlja se, da ima pri preživetju v naravnem okolju pomembno vlogo tvorba biofilma. Prvi in ključni korak, potreben za uspešno tvorbo biofilma, je adhezija bakterij na ustrezno površino. V magistrskem delu je bila ovrednotena sposobnost adhezije in invazije bakterij Campylobacter jejuni na/v humane črevesne celice karcinogenega izvora Caco-2. V raziskavo so bili vključeni referenčni sev Campylobacter jejuni (divji sev NCTC 11168) in njegovi mutanti z mutacijami v genih chuA, csrA, kpsM, luxS, omp50, rrpA, rrpB, trpE in Cj1191c. Vpliv mutacij v izbranih genih na sposobnost adhezije in invazije bakterij Campylobacter jejuni je bil ovrednoten z metodo štetja kolonij (CFU/ml). Pred tem je bil ovrednoten vpliv mutantov na metabolno aktivnost celic Caco-2 s kolorimetričnim testom z uporabo reagenta MTT. Statistična analiza je pokazala, da so mutacije v genih chuA, csrA, luxS, omp50 in rrpB statistično značilno zmanjšale sposobnost adhezije bakterij Campylobacter jejuni na celice Caco-2. Mutacije v genih kpsM, trpE in Cj1191c so statistično značilno povečale sposobnost adhezije bakterij Campylobacter jejuni na celice Caco-2. Sposobnost invazije v celice Caco-2 sta imela samo mutanta z mutacijo v genu rrpB in genu luxS. Ostali mutanti niso invadirali v celice Caco-2. Potrebne so nadaljnje raziskave za razvoj strategij, ki omejujejo število okužb in omogočajo nadzor bakterij v živilih.
Keywords:	Campylobacter jejuni, tvorba biofilma, adhezija, invazija, celice Caco-2
Place of publishing:	Maribor
Place of performance:	Maribor
Publisher:	[T. Damijan]
Year of publishing:	2022
Number of pages:	1 spletni vir (1 datoteka PDF (XII, 79 str.))
PID:	20.500.12556/DKUM-83462
UDC:	579:579.8:628.353.153(043)=163.6
COBISS.SI-ID:	136101891
Publication date in DKUM:	04.01.2023
Views:	871
Downloads:	104
Metadata:
Categories:	FKBV
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Licences

License:	CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International

Link:	http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:	The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:	30.11.2022

Secondary language

Language:	English
Title:	The influence of biofilm-relevant gene mutations on the adhesion and invasion ability of campylobacter jejuni in caco-2 cells
Abstract:	Despite its difficult growth conditions, the microaerophilic bacterium Campylobacter jejuni is a widespread microorganism and the most common cause of acute bacterial gastroenteritis in humans worldwide. Biofilm formation is thought to play an important role in its survival in natural environments. The first and most important step for the successful formation of a biofilm is the adhesion of bacteria to a suitable surface. In this master thesis, the adhesion and invasion ability of Campylobacter jejuni bacteria on/in carcinogenic human intestinal cells Caco-2 was evaluated. The reference strain Campylobacter jejuni (wild strain NCTC 11168) and its mutants with mutations in the genes chuA, csrA, kpsM, luxS, omp50, rrpA, rrpB, trpE and Cj1191c were included in the study. The effects of mutations in selected genes on the adhesion and invasion ability of Campylobacter jejuni were evaluated using the colony count method (CFU/ml). First, the influence of the mutations on the metabolic activity of Caco-2 cells was assessed using a colorimetric MTT assay. Mutations in the genes chuA, csrA, luxS, omp50 and rrpB statistically significantly decreased the adhesiveness of Campylobacter jejuni bacteria to Caco-2 cells. In contrast, mutations in the kpsM, trpE and Cj1191c genes statistically significantly increased the adhesion ability of Campylobacter jejuni bacteria to Caco-2 cells. Only one mutant with a mutation in the rrpB gene and one mutant with a mutation in the luxS gene had invasive abilities in Caco-2 cells. The other mutants did not invade Caco-2 cells. Further research is needed to develop strategies to limit the number of infections and control bacteria in food.
Keywords:	Campylobacter jejuni, biofilm formation, adhesion, invasion, Caco-2 cells

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