ISBN 978-82-326-5580-9 (printed ver.) ISBN 978-82-326-6774-1 (electronic ver.) ISSN 1503-8181 (printed ver.) ISSN 2703-8084 (online ver.)
Doctoral theses at NTNU, 2021:204
Kasper Schei
Early Gut Fungal and Bacterial Microbiota in Growth and
Allergy-Related Diseases
Doctor al thesis
Doctoral theses at NTNU, 2021:204Kasper Schei NTNU Norwegian University of Science and Technology Thesis for the Degree of Philosophiae Doctor Faculty of Medicine and Health Sciences Department of Clinical and Molecular Medicine
Thesis for the Degree of Philosophiae Doctor Trondheim, June 2021
Norwegian University of Science and Technology Faculty of Medicine and Health Sciences
Department of Clinical and Molecular Medicine
Kasper Schei
Early Gut Fungal and Bacterial Microbiota in Growth and
Allergy-Related Diseases
NTNU
Norwegian University of Science and Technology Thesis for the Degree of Philosophiae Doctor Faculty of Medicine and Health Sciences Department of Clinical and Molecular Medicine
© Kasper Schei
ISBN 978-82-326-5580-9 (printed ver.) ISBN 978-82-326-6774-1 (electronic ver.) ISSN 1503-8181 (printed ver.)
ISSN 2703-8084 (online ver.) Doctoral theses at NTNU, 2021:204 Printed by NTNU Grafisk senter
T
IDLEG SOPP-
OG BAKTERIEMIKROBIOTA I VEKST OG ALLERGIRELATERTE SJUKDOMMARB
ARNETARMFLORA I VEKST-
OG ALLERGIUTVIKLINGDet er ikkje tvil om at tarmflora er i vinden for tida. Endeleg har forskarar kunna djupdukke ned i mikrobemiljøet i tarmen, for no har ein fått mye betre metodar å gjere det på. Tarmfloraen er det gløymte organet fordi ein har visst om det lenge, men først dei siste åra har forstått at desse 1-2 kiloa av tarmbakteriar, tarmsoppar og andre tarmmikrobar faktisk påverkar oss.
Til no har ein sett mest på korleis bakteriar i tarmen påverkar oss, men hos dei aller minste barna er dette ikkje fullt utforska. Om tarmsoppen, som berre utgjer ein brøkdel av tarmmikrobane, veit ein enda mindre, og knapt noko hos barn. Denne avhandlinga omfattar tre studiar kor den første har skildra barnesoppfloraen hos barn frå 10 dagar til 2 år og mødrene deira. I studie to og tre har ein sett på om det er samanheng mellom sopp- og bakterieflora hos desse barna, med høvesvis vekstutvikling fram til 9 år og astma- og allergiutvikling fram til 6 år. Vi har analysert resultat frå 298 (studie 1) og 278 barn (studie 2 og 3) og deira mødrer frå ProPACT-studien som vart gjennomført i Trondheim på starten av 2000-talet. Der målte vi mengda sopp- og bakterie-DNA, og vi prøvde å finne ut kva for sopp- og bakterieartar det DNA-et kom frå ved å sekvensere det.
I første studie såg vi at 88 % av mødrer og 56-76 % av barna hadde påviseleg sopp i
avføringsprøvene sine. Litt usikkerheit rundt utvinninga av sopp-DNA-et gjer at det kan vere meir. Dessutan hadde barn høgare sjanse for å ha påviseleg tarmsopp om mor hadde det. Den vanlegaste tarmsoppen hos spedbarna var Debaryomyces hansenii, ei mjølkegjær som òg finst i Jarlsberg. Hos dei større barna var brødgjæra vanlegast (Saccharomyces cerevisiae). Vi klarte berre å finne soppartar hos 4 % av barna, som gjer at desse resultata er usikre.
I andre og tredje studie fann vi at toåringane med mye tarmsopp vart høgare mellom 2-9 år, og desse hadde òg høgare sjanse for å utvikle astma og høysnue fram til 6 år. Dessutan fann vi at meir sopp og bakteriar i tarmen hos eittåringar var tynnare første leveåret. Vi har med dette funne ein samanheng, men det tyder ikkje direkte at ein blir høgare og allergisk av å ete ein godt gjæra bolledeig. Ein har likevel føreslått at visse korte feittsyrer og hormon han henge i hop med vekst- og astmautviklinga.
Dette er heilt nye eksperimentelle funn som ein treng å stadfeste i nye studiar for å kunne stole fullt på dei. Likevel peiker dei på at tarmfloraen, og særleg tarmsoppen, kan ha innverknad på den utviklande kroppen til barna som veks opp blant oss.
Kandidat: Kasper Schei
Institutt: Institutt for klinisk og molekylær medisin (IKOM)
Rettleiarar: Rønnaug Ødegård (hovudrettleiar), Saideh Salamati, Torbjørn Øien, Knut Rudi Finansieringskjelder: NTNU, Noregs Forskingsråd, TINE AB
Den ovannemnde avhandlinga er funnen verdig for offentleg forsvar for graden philosophiae doctor (ph.d.) i medisin.
Disputasen finn stad digitalt 11. juni 2021.
TABLE OF CONTENTS
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A CKNOWLEDGEMENTS
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L IST OF PUBLICATIONS
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A BBREVIATIONS
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1 I NTRODUCTION
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MICROBIOME METHODS
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?PG<IM8I@89C<I<>@FEJKFNFIB8J98I:F;<J=FI<8:?JG<:@<J.@E:<98:K<I@88I<GIFB8IPFK<J K?<@I):F;<J=FI898:K<I@8 FECP.JL9LE@KF=K?<I@9FJFD<!@>LI< LB8IPFK<JC@B<
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Figure 1. Illustation of the ribosomal DNA used as identity markers in metabarcoding. /?<>I<<E9FO<J J?FNK?<):F;@E>=FIK?<JL9LE@KJ@EK?<I@9FJFD<J8E;K?<J<8I<;@==<I<EK@E<LB8IPFK<J8E;GIFB8IPFK<J 1 18I<K?<?PG<IM8I@89C<I<>@FEJF=K?<..I@9FJFD<)J<HL<E:</?<$EK<IE8CKI8EJ:I@9<;JG8:<IJ
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FUNGAL MICROBIOTA IS CALLED MYCOBIOTA
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@J8:C8JJ@=@:8K@FE<EK@KPC@B<8JG<:@<JFI8><ELJ
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&6,"3:05*$FI>8E@JDJ"I<<B&6dhN<CC>FF;i,"3:0/dhB<IE<C:FI<i9<:8LJ<K?<=LE>8C :<CC?FC;J8EL:C<LJ@EN?@:?K?<D8AFI@KPF=K?<:<CCiJ><EFD<@J/?<=LE>8C><EFD<
:FEK8@EJD8@ECP@EKI8EL:C<8I)8II8E><;@E?8GCF@;:?IFDFJFD<JF=M8I@FLJELD9<IJ
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:8EEFK:FEM<IK<E<I>P=IFDC@>?K@EG?FKFJPEK?<J@J8E;8I<K?<I<=FI<;<G<E;<EKFE89JFI9@E>
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P<8JK:<CCJ:8CC<;GJ<L;F?PG?8<J<<E@E8E;@;8JGG(FLC;JFEK?<FK?<I?8E;>IFN@E JKI@E> C@B<JKIL:KLI<J:8CC<;?PG?8</?<J<?PG?8<=FID8D8:IFJ:FG@:IFFKJKIL:KLI<:8CC<;
DP:<C@LD8E;K?@JB@E;F=>IFNK?@JJ<<E@E41&3(*--64JGG.FD<F=K?<DFJKG8K?F><E@:
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9FK?P<8JK8E;DFLC;J:8EI<GIF;L:<8J<OL8CCPN?<I<E<N=LE>8CJGFI<J8I<D8;<DP D@KFJ@JFIK?IFL>?J<OL8C;@M@J@FEN?<I<KNF:<CCJLE;<I>FD<@FJ@J@EJGFI<=FID8K@FE 0EBEFNE=LE>@:FEK@EL<KF9<;@J:FM<I<;8K8?@>?I8K<8E;:8C:LC8K@FEJJ?FNK?8KFECP 89FLKF=8CC=LE>@J<<DKF?8M<9<<E=FLE;@DGCP@E>8KFK8CELD9<IF= D@CC@FE
=LE>8CJG<:@<JFLKK?<I<'@B<98:K<I@8=LE>@8I<L9@HL@KFLJ@EFLI<EM@IFED<EK9<@E>
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?LD8E?@JKFIPK?<@I89@C@K@<JN<I<;@J:FM<I<;<8ICP8E;<OGCF@K<;@E?LD8E=FF;:LCKLI<C@B<
9I<8;8C:F?FC@:9<M<I8><I@G<E<;:?<<J<8E;FK?<I=<ID<EK<;GIF;L:KJ/?<;@J:FM<IPF=
G<E@:@CC@E8E8EK@98:K<I@8CGIF;L:KF=K?<8@I 9FIE<DFLC;&/*$*--*6.N8J8D<;@:8CKLIE@E>
GF@EKCJFK?<;@J:FM<IPF==LE>@8JGFK<EK@8C?LD8EG8K?F><EJI<MFCLK@FE@J<;:C@E@:8C D<;@:@E<
Gut mycobiota
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/?<=LE>@K?8KGFGLC8K<K?<>8JKIF@EK<JK@E8CKI8:K8I<:8CC<;K?<(65.:$0#*05""I<<B.:,&4d h=LE>LJi#*05"dhC@=<i/?<>LKDP:F9@FK8?8JI<:<EKCP9<<EI<:F>E@J<;8J8JL9JK8EK@8CG8IK F=K?<>LKD@:IF9@FK88E;:8E9<@DGFIK8EK=FI?LD8E?<8CK?/?<>LKDP:F9@FK8N8J
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@E;<G<E;<EKD<K?F;JN<I<;<M<CFG<;0J@E>8:LCKLI< @E;<G<E;<EKD<K?F;:8CC<;J?FK>LE J<HL<E:@E>F=K?<8;LCK>LKD@:IF9@FD<FE<?8JJ?FNEK?8KFECPF=K?<D@:IF9@8C )@J=LE>8CCK?FL>?K?@JG<I:<EK8><@J;<98K<;8E;D@>?K9<LE;<I<JK@D8K<;>LK=LE>@
8I<G8IKF=K?<>LKJ3"3&#*041)&3&D@:IF9@8CJG<:@<JF=CFNHL8EK@KP@E8E<:FJPJK<D )<M<IK?<C<JJ9<@E><LB8IPFK<J=LE>8C:<CCJ8I<89FLKbD@E;@8D<K<IN?<I<8J98:K<I@8 I<8:?89FLK bD8:IFJJ8CK?FL>?K?<I<@JJ@>E@=@:8EKM8I@8K@FE@EJ@Q<JJLD@E>K?<
MFCLD<F=8JG?<I<
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JPJK<D8E;@KJI<JGFEJ<JP8:K@M8K@E>K?<=LE>LJ JG<:@=@:G8K?F><E I<:F>E@K@FEI<:<GKFIJ +--J8E;8;A8:<EKD<:?8E@JDJ;<=<E:<J8>8@EJK?8ID=LCG8K?F><EJ8E;KFC<I8E:<KFN8I;J
?<CG=LC:FDD<EJ8CJ8I<=FID<;
FUNGAL AND BACTERIAL GUT MICROBIOTA IN CHILDREN
Microbiota diversity
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@J:FDDFECP<OGI<JJ<;@E.?8EEFE @E;<O#iFI.@DGJFEiJI<:@GIF:8C@E;<O /?<#&5"%*7&34*5:o;@M<IJ@KP<OGI<JJ<J?FN;@M<IJ<KNFJ8DGC<J8GG<8I@EK<IDJF=
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Foetal microbiota
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GUT PERMEABILITY AND INFLAMMATION
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3 A IMS
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4 M ATERIALS AND METHODS THE STUDY POPULATION
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Table 1A. Participant characteristics in Paper I (N = 298) (8K<IE8C8><8K;<C@M<IPP<8IJ.
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Table 1B. Participant characteristics in Paper II and III (N = 278)
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