magistrsko delo
Anja Ošep (Author), Maja Rupnik (Reviewer), Marjanca Starčič Erjavec (Mentor), Valerija Zidarič (Co-mentor)

Abstract

Sestava črevesne mikrobiote se od posameznika do posameznika razlikuje. V zadnjem času se raziskovalci vse bolj osredotočajo na proučevanje sporogenih predstavnikov črevesne mikrobiote in njihovo vlogo pri horizontalnem prenosu med ljudmi. V tovrstnih raziskavah največjo težavo predstavlja identifikacija sporogenih bakterij v črevesni mikrobioti. Raziskave črevesne mikrobiote ponavadi temeljijo na metagenomskem sekvenciranju, ki ima kljub mnogim prednostim tudi pomanjkljivosti, saj vseh organizmov s to metodologijo ne zaznavamo enako dobro. Z namenom odprave tovrstnih omejitev se vse bolj razvija kulturomika. Namen naše naloge je bil osamitev in identifikacija čim večjega števila sporogenih bakterij iz fecesa človeka ter njihova primerjava med posameznimi družinskimi člani. Uporabili smo vzorce fecesa štirih zdravih prostovoljcev, ki živijo v istem gospodinjstvu. Vzorce fekalnih suspenzij smo obdelali z etanolom in jih cepili na različna trdna gojišča. Vzporedno smo vzorce fekalnih suspenzij predhodno gojili tudi v tekočih gojiščih in jih nato cepili na trdna gojišča enake sestave. Za identifikacijo sporogenih predstavnikov črevesne mikrobiote smo uporabili molekularne in gojitvene metode. Za identifikacijo čistih kultur smo uporabili MALDI TOF MS in sekvenciranje gena za 16S rRNA, celotno združbo bakterij pa smo analizirali s 16S rRNA metagenomskim sekvenciranjem. Ugotovili smo, da predhodno gojenje vzorcev fekalne suspenzije v tekočih gojiščih ni prispevalo k identifikaciji večjega nabora predstavnikov črevesne mikrobiote. Na podlagi analize osamljenih izolatov smo opazili prekrivanje posameznih bakterijskih vrst med družinskimi člani. Pri proučevanih družinskih članih smo opazili tudi prekrivanje posameznih ZOTU sporogenih sevov.

Keywords

črevesna mikrobiota;sporogene bakterije;vzorci fecesa;kultivacija;metagenomika;družinski člani;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [A. Ošep]
UDC: 579.25:575.111:616.34
COBISS: 102444547 Link will open in a new window
Views: 279
Downloads: 75
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Other data

Secondary language: English
Secondary title: Comparison of spore-forming bacteria from the intestinal microbiota of cohabiting family members
Secondary abstract: The composition of gut microbiota varies between individuals. Recently, researchers are focusing on the studies of spore-forming bacteria from the human gut microbiota and their role in host to host transmission. In such studies, the main problem is the identification of spore-forming bacteria from the gut microbiota. Studies on the human gut microbiota are usually based on metagenic sequencing. Despite its many advantages this method also has limitations as not all microorganisms are equally well detected. In order to reduce these limitations the culturomics has been developing. The aim of our master thesis was to isolate and identify spore-forming bacteria from fecal samples and to compare them between cohabiting family members. Stool samples from four healthy individuals were used. Stool samples were treated with alcohol and inoculated on different solid media. Stool samples had also been cultivated in liquid media and then inoculated on different solid media. To identify spore-forming bacteria from human gut microbiota molecular and culture methods were used. MALDI TOF MS and 16S rRNA gene sequencing were used to identify pure cultures, and the whole bacterial community was analyzed by 16S rRNA metagenomic sequencing. We found that cultivation of faecal suspension in liquid media did not increase the number of identified bacteria from human gut microbiota. Analysis of the pure cultures showed that some bacterial species were present in all family members. Moreover, some ZOTU of the spore-forming strains were present in all family members.
Secondary keywords: gut microbiota;spore-forming bacteria;fecal samples;cultivation;metagenomics;cohabiting family members;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Oddelek za mikrobiologijo
Pages: XIII, 81 f., [7] f. pril.
ID: 14851234