Zebrafish colonization with human intestinal bacteria DOI: 10.1089/zeb.2017.1460

1. Zebrafish gut as a house
for human intestinal bacteria
Nerea Arias Jayo
PhD student
AZTI-Tecnalia,Food Research Division
4th World Congress on Targeting Microbiota, Paris 2016

2. Zebrafish model
• Widely use
• Genetically similar to
humans
• Easy and cheap to house
and care
. . .
1
Human gut microbiota:
• Complex
• High variability
Gnotobiotic animals
www.Acuariored.com
Introduction

3. • Divided in 3 parts
• ¾ mammalian principal
intestinal c.:
– Absorptive E.
– Goblet C.
– Enteroendocrine C.
Introduction
Zebrafish intestine
https://www.researchgate.net/publication/7635932_Formation
_of_the_digestive_system_in_zebrafish_III [accessed Oct 15,
2016]

4. • The intestinal
wall folds
resembling the
mammal villis
• ZF has not
submucosa layer
K.N. Wallace et al. / Mechanisms of Development 122 (2005) 157–173
Introduction
Zebrafish intestine

5. • Zf microbiota is
different from
humans
• MAMPS –
TLR/NLR
D. Alexandar et al. / Genes Dev. 2013 27: 701-718
Introduction
Zebrafish intestine

6. E. Coli E. faecalis L. casei B. breve E. limosum
Materials & methods
1 • Colonization of 5dpf zebrafish larvae
with 5 human gut bacteria
2 • Colonization 5 dpf zebrafish larvae
with human intestinal microbiota
meetgenes.blogs.uv.es

7. Materials & methods 1
Embryo collection
Gnotobiotic protocol
Infection (48h)
(Facultative anaerobes:
E.coli, E. faecalis, L. casei;
Obligate anaerobes:
B. breve and E. limosum)
– Monoassociation
– Consortia
Colonization confirmation
0 dpf
5 dpf
24,48 hpi, 7,10dpi
Germ-free
1 • Colonization 5dpf zebrafish larvae with 5 human gut bacteria

8. Materials & methods 2
Embryo collection
Gnotobiotic protocol
Infection (48h)
Colonization confirmation
by:
- RISA
- Metagenomics
0 dpf
5 dpf
7 dpf (48 hpi)
2 • Colonization 5 dpf zebrafish larvae with human microbiota
Germ-free

9. Results and discussion
Consortia
- Facultative anaerobes are able to survive
up to 10 days
- Obligate anaerobes only survive 48 hours
- E. faecalis and E. coli increase their
growth with time
The oxygen pressure makes
possible facultative anaerobe
growth, limiting obligate
anaerobe survival.
Monoassociations
- Higher counts for all the species used
Probably due to no interspecific
competition.
All the strains remain inside the
zebrafish gut almost 48 hours
• Colonization 5dpf zebrafish larvae with 5 human gut bacteria
Fig 1: Comparison of E. coli, E.
faecalis, L. casei, B. breve, E.
limosum colonization in
monoassociation ( MA ) and in
consortia ( C )

10. Conclusion
Developing zebrafish could be a suitable model for studying:
• The human gut microbiota
• Interactions with the host
•Microbiota dysbiosis in diseases (diabetes, obesity, IBD…)
Some human intestinal bacteria, including obligate anaerobes,
are able to colonize and compete inside the zebrafish gut
Successfully
colonized
5 dpf
zebrafish gut
5 human gut
bacterial sp

11. To the Department of Environment,
Territorial Planning, Agriculture and
Fisheries of the Basque Country
Government for the scholarship
LIFE13 ENV/ES/001048
This research is part of Enviphage project
(www.envihage.eu) and is financed by the
LIFE+ environmental programme of the
European Commission
Acknowledgments
Herrera kaia, Portualdea z/g
20110 Pasaia, Gipuzkoa
Astondo bidea, Edificio 609
Parque Tecnológico de Bizkaia
48160 Derio, Bizkaia
Txatxarramendi ugartea z/g
48395 sukarrieta, Bizkaia