1. Bacterial and Archaeal Communities in the Brine/Seawater Interface of Four Different Brine Lakes in the Mediterranean Sea. Henk Bolhuis Paul van der Wielen Laboratory of Microbial Ecology Center for Ecological and Evolutionary Studies University of Groningen The Netherlands Laboratory of Microbial Ecology

3. Questions What type of bacteria and archaea dominate the brine interface in four different brines? Have the bacterial and archaeal communities in the four different brine interfaces developed in a similar manner? Laboratory of Microbial Ecology

4. Brine interface water Filter DNA-isolation Bacterial 16S rDNA PCR Archaeal 16S rDNA PCR Cloning PCR- product Sequencing 700 bases Cloning PCR- product Laboratory of Microbial Ecology

5. BrineGroupSequencesOTUsShannon Atalante Bacteria 96302.962 Bannock Bacteria 96292.774 Discovery Bacteria 96383.329 Urania Bacteria 96292.862 Atalante Archaea 93152.010 Bannock Archaea 96141.789 Discovery Archaea 96151.789 Urania Archaea 96151.463 Brine interface diversity Laboratory of Microbial Ecology

6. AtalanteBannockDiscoveryUraniaX Atalante100%38.5%18.8%26.0%42.7% Bannock54.1%100%14.6%52.1%24.0% Discovery12.5%10.4%100%9.4%79.2% Urania31.3%28.1%20.8%100%43.8% Bacterial brine interface similarity (OTU-level) Laboratory of Microbial Ecology

7. AtalanteBannockDiscoveryUraniaX Atalante100%38.7%95.7%93.5%1.1% Bannock81.3%100%78.1%82.3%14.6% Discovery83.3%13.5%100%80.2%15.6% Urania93.4%78.1%92.7%100%4.2% Archaeal brine interface similarity (OTU-level) Laboratory of Microbial Ecology

9. AtalanteBannockDiscoveryUrania  -proteobacteria 3211  -proteobacteria 4192216  -proteobacteria 40025  -proteobacteria 12220 Dehalococcoides0200 CFB-group83133 Fibrobacter/Acidobacterium1501 Verrucomicrobia1440 Actinobacteria2560 Planctomycetales0221 Haloanaerobiales0020 Candidate OP94705 Candidate OP110032 Candidate KB14010 New 134282 New 24657 New 37214 New 47802 New 59090 New 62000

10. Laboratory of Microbial Ecology

11. AtalanteBannockDiscoveryUrania Marine group I359666 Methanosarcinales5031 Halobacteriales00111 Thermoplasmatales0001 Candidate DHVE70100 New 10700 New 211072 New 30202 New 474236923 New 50400 Laboratory of Microbial Ecology

12. Conclusions Interface bacterial communities are brine specific Interface archaeal communities are not brine specific Laboratory of Microbial Ecology

13. The bacterial community in the Discovery brine interface differs considerably from the other three brine interfaces At group level Atalante and Discovery are different from Bannock and Urania Conclusions Laboratory of Microbial Ecology

14. Acknowledgements Groningen Jolanda Brons BioDeep EU-Fifth framework Contract EVK3-2000-22057 Milano Sara Borin Daniele Daffonchio Europe BioDeep-partners Laboratory of Microbial Ecology www.geo.unimib.it/BioDeep/project