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

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

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

BrineGroupSequencesOTUsShannon Atalante Bacteria Bannock Bacteria Discovery Bacteria Urania Bacteria Atalante Archaea Bannock Archaea Discovery Archaea Urania Archaea Brine interface diversity Laboratory of Microbial Ecology

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

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

AtalanteBannockDiscoveryUrania  -proteobacteria 3211  -proteobacteria  -proteobacteria  -proteobacteria Dehalococcoides0200 CFB-group83133 Fibrobacter/Acidobacterium1501 Verrucomicrobia1440 Actinobacteria2560 Planctomycetales0221 Haloanaerobiales0020 Candidate OP94705 Candidate OP Candidate KB14010 New New New New New New 62000

Laboratory of Microbial Ecology

AtalanteBannockDiscoveryUrania Marine group I Methanosarcinales5031 Halobacteriales00111 Thermoplasmatales0001 Candidate DHVE70100 New New New New New Laboratory of Microbial Ecology

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

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

Acknowledgements Groningen Jolanda Brons BioDeep EU-Fifth framework Contract EVK Milano Sara Borin Daniele Daffonchio Europe BioDeep-partners Laboratory of Microbial Ecology