Kolonizacijski potencijal i učinak na intestinalnu mikrobiotu Lactobacillus plantarum SF9C i Lactobacillus brevis SF9B in vivo

master's thesis

• Title (english): Colonization potential and effect on intestinal microbiota of Lactobacillus plantarum SF9C and Lactobacillus brevis SF9C in vivo
• Author: Teo Vurnek
• Mentor: Jasna Novak (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Antibiotic, Enzyme, Probiotic and Starter Cultures Technology); Zagreb
• Defense date and country: 2019-06-17, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Cilj ovog diplomskog rada je istražiti kolonizacijski potencijal i učinak na sastav intestinalne mikrobiote sojeva Lactobacillus brevis SF9B koji sintetizira S-protein i Lactobacillus plantarum SF9C producenta plantaricina, koji se istražuju kao probiotički sojevi, in vivo na modelu eksperimentalnih štakora. Za analizu sastava intestinalne mikrobiote korišteni su različiti pristupi, PCR-DGGE metoda za detekciju prisutnosti odabranih Lactobacillus sojeva, nakon prolaska kroz gastrointestinalni trakt (GIT) in vivo, dok je najnovija generacija sekvencioniranja primjenjena za određivanje redoslijeda nukleotida V3 i V4 varijabilnih regija 16S rRNA gena mikrobioma pomoću Illumina MiSeq platforme s ciljem analize sastava i učinka odabranih sojeva na intestinalnu mikrobiotu. PCR-DGGE metodom potvrđeno je da oba ispitivana soja laktobacila preživljavaju u visokom broju kolonija nakon prolaska kroz GIT, 3 i 10 dana nakon prestanka primjene, a ustanovljena je i prisutnost drugih Lactobacillus vrsta, koje su dio autohtone populacije: Lactobacillus animalis, Lactobacillus intestinalis i Lactobacillus reuteri. Metagenomskom analizom potvrđena je bioraznolikost Lactobacillus spp. u mikrobioti štakora u kojima su aplicirani SF9C i SF9B u usporedbi s kontrolnom skupinom i 10 dana nakon završetka tretmana. Rezultati ovog istraživanja doprinose hipotezi o funkcionalnosti sojeva SF9C i SF9B. Združena primjena dva soja, doprinosi adhezijskim svojstvima Lb. plantarum SF9C uslijed kompetitivne ekskluzije patogena djelovanjem Lb. brevis SF9B, a soj SF9B ima pojačan kolonizacijski potencijal kojem doprinosi antimikrobna aktivnost plantaricina Lb. plantarum SF9C, koji iskazuje širi spektar antimikrobnog djelovanja uključujući Listeria monocytogenes ATCC 19111.

Abstract (english)

The aim of this work is to investigate the colonization potential and effect on the composition of the intestinal microbiota of S-layer carrying Lactobacillus brevis SF9B and plantaricin producing Lactobacillus plantarum SF9C in vivo on the model of experimental rats. Two different approaches were applied for the analysis of the intestinal microbiota composition. PCR-DGGE method was employed for assessing the presence of selected Lactobacillus strains, after their transit through the gastrointestinal tract (GIT) of the rat, while next-generation sequencing via Illumina MiSeq platform was applied for determination intestinal microbiota composition and effect of the selected strains in the, by amplifying V3 and V4 16S rRNA gene regions. PCR-DGGE analysis revealed the presence of both examined strains as well as detection of other Lactobacillus species representatives, the identified as Lactobacillus animalis, Lactobacillus intestinalis and Lactobacillus reuteri, which are autochthonous bacteria in the rat intestine, even 3rd and 10th day after the end of application. The metagenomic analysis confirmed an abundant prevalence of Lactobacillus spp. was in the gut microbiota of rats inoculated with SF9C and SF9B strains compared with the control group, even 10 days after the end of treatment. The results of this research supported enhanced functionality potential of jointed application of SF9C and SF9B strains, where both strains can benefit, Lb. plantarum SF9C by facilitated adhesion due to the competitive pathogen exclusion by coexisting Lb. brevis SF9B, while in same SF9B benefits in improved colonization due to the plantaricin production by Lb. plantarum SF9C, resulting altogether in a broader spectrum of antimicrobial activity against pathogens of the coculture.

• Keywords: intestinalna mikrobiota; kolonizacija in vivo; sekvencioniranje; Lactobacillus
• Keywords (english): intestinal microbiota; in vivo colonization; next-heneration sequencing; Lactobacillus
• Language: croatian
• URN:NBN: urn:nbn:hr:159:433507
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2019-12-06 13:36:20