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Microbial Diversity in Azorean Lava Tubes
Riquelme, C.; Hathaway, J.M.M. ; Gabriel, R. ; Louvado, A. ; Coelho, M. ; Dapkevicius, M.L.N.E. & Northup D.E.
II EIBS, Aveiro 2nd September 2011
Microbial Mats Diversity in Azorean Lava Tubes
© Kenneth Ingham Photography
© Kenneth Ingham Photography
© Kenneth Ingham Photography
© Kenneth Ingham Photography
© Kenneth Ingham Photography
1 m 1 cm
1 cm
1 cm 1 cm
II EIBS, Aveiro 2nd September 2011
© Kenneth Ingham Photography © Kenneth Ingham Photography
© Kenneth Ingham Photography
© Kenneth Ingham Photography
© Kenneth Ingham Photography © Kenneth Ingham Photography
Microbial Mats Diversity in Azorean Lava Tubes
5 mm
Microorganisms live in community, biofilms
II EIBS, Aveiro 2nd September 2011
20 mm
© Diana Northup Photography
Lava Tubes –Extreme Environments
• Scarcity of energy for life – Light (rely on chemical energy) – Nutrients (have to produce organic compounds from
inorganic molecules)
II EIBS, Aveiro 2nd September 2011
• Low temperatures
• Microbial diversity is fundamental to maintenance and conservation of global genetic resources – Huge amount of Earth biomass
– Genetic flexibility that allows them to adapt to (almost) every environmental conditions
• Extreme Environments as a diverse and untapped biodiversity refuge
• Earth Caves As Analogs For Extraterrestrial Cave Life Detection
On The Importance Of Studying Microorganisms in Lava Tubes
II EIBS, Aveiro 2nd September 2011
Microorganism effects on cave formation, enlargement, & mineral precipitation
Who’s home? Is there an indigenous subsurface biota?
Do microorganisms adapt to the cave environment? Do they produce antimicrobials?
Themes In Cave Microbiology
Buracos cave, Terceira, Portugal (Kenneth L. Ingham) II EIBS, Aveiro 2nd September 2011
Lava Tubes in Azorean Archipelago
TERCEIRA 69 CAVES
PICO 129 CAVES
FLORES 3 CAVES
FAIAL 9 CAVES
SÃO JORGE 19 CAVES
GRACIOSA 10 CAVES
SÃO MIGUEL 28 CAVES
SANTA MARIA 4 CAVES
CORVO
http://www.speleoazores.com/ II EIBS, Aveiro 2nd September 2011
Azorean Lava Tubes
Terceira Balcões
Malha
Terra Mole
Buracos
Madre de Deus
Agulhas
Achada
Branca Opala
Principiantes
Natal
Algar do Carvão
Pico Montanheiros Torres Lemos Ribeira do Fundo
II EIBS, Aveiro 2nd September 2011 © Kenneth Ingham Photography
Studies on the microbial biodiversity in the lava tubes of the Azores
Nucleic acid analysis by metagenomics (culture-independent techniques)
Culture in lab mimicking cave environment
II EIBS, Aveiro 2nd September 2011
© Kenneth Ingham Photography © Kenneth Ingham Photography © Kenneth Ingham Photography
Metagenomics
Molecular identification of bacterial isolates by sequencing, previously inaccessible since it is estimated that only less than 5% of the total bacterial species can be grown in the lab
Molecular ecology studies with analysis of entire communities of bacteria
Estimation of microbial diversity, which allows us to establish strategies to increase the proportion of the cultivable microflora cave
Metagenomes, genetic material sampled directly from the environment
II EIBS, Aveiro 2nd September 2011
Scientific classification of organisms
II EIBS, Aveiro 2nd September 2011
Ciccarelli, F.D., et al., Toward Automatic Reconstruction of a Highly Resolved Tree of Life, 2006 Vol 311 Science
ARCHAEA
EUCARYA
BACTERIA
II EIBS, Aveiro 2nd September 2011
Closer view of some evolutionary distances
METAZOA
FUNGI
CHROMALVEOLATA
PLANTAE
Ciccarelli, F.D., et al., Toward Automatic Reconstruction of a Highly Resolved Tree of Life, 2006 Vol 311 Science
II EIBS, Aveiro 2nd September 2011
Scientific classification of organisms Evolutionary distance
ARCHAEA
EUCARYA
BACTERIA
Ciccarelli, F.D., et al., Toward Automatic Reconstruction of a Highly Resolved Tree of Life, 2006 Vol 311 Science
Acidobacteria Actinobacteria Aquificae Bacteroidetes Chamydiae Chlorobi Chloroflexi Chrysiogenetes Cyanobacteria Deferribacteres Dictyoglomi Fibrobacteres Firmicutes Fusobacteria Gemmatimonadetes Lentisphaerae Nitrospirae Planctomycetes Proteobacteria Spirochaetes Synergistetes Thermi-Deinococci Thermodesulfobacteria Thermotogae Verrucomicrobia
Candidate Bacterial Phyla
II EIBS, Aveiro 2nd September 2011
GN02 OD1 OP3 OP9 OP10 OP11 SR1 TG1 TM7 WS3 WS6 Ktenodobacteria
Norman R. Pace; Mapping The Tree Of Life: Progress And Prospects Microbiology And Molecular Biology Reviews, Dec. 2009, P. 565–576
Cavaletti, L., Monciardini, P., Bamonte, R., Schumann, P., Rohde, M., Sosio, M. & Donadio, S. (2006). New lineage of filamentous, spore-forming, gram-positive bacteria from soil. Appl Environ Microbiol 72, 4360–4369.
8 10
5
9 12
9 7
10 10
Phyla recovered by Cave
Torres
Ribeira do Fundo
Principiantes
Montanheiros
Malha
Lemos
Branco Opala
Balcões
Achada
II EIBS, Aveiro 2nd September 2011
ACIDOBACTERIA 15,3
ACTINOBACTERIA 14,0
KTEDONOBACTERIA 0,2
BACTEROIDETES 2,3
CHLAMYDIAE 0,1
CHLOROFLEXI 2,8
CYANOBACTERIA 1,5
FIRMICUTES 3,1
GEMMATIMONADETES 1,7
NITROSPIRA 8,0
OP10 0,4
PLANCTOMYCETES 0,6
PROTEOBACTERIA UNCLASSIFIED
1,5
PROTEOBACTERIA-ALPHA
12,7 PROTEOBACTERIA-BETA
6,6
PROTEOBACTERIA-DELTA
4,4
PROTEOBACTERIA-GAMMA
12,1 TM7 0,1 UNCLASSIFIED
12,4
VERRUCOMICROBIA 0,4
II EIBS, Aveiro 2nd September 2011
Percentages of phyla recovered in Terceira and Pico Caves
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Terceira Pico
UNCLASSIFIED
TM7
VERRUCOMICROBIA
PLANCTOMYCETES
OP10
CYANOBACTERIA
KTEDONOBACTERIA
NITROSPIRA
GEMMATIMONADETES
FIRMICUTES
CHLAMYDIAE
CHLOROFLEXI
BACTEROIDETES
ACTINOBACTERIA
ACIDOBACTERIA
PROTEOBACTERIA-GAMMA
PROTEOBACTERIA-DELTA
PROTEOBACTERIA-BETA
PROTEOBACTERIA-ALPHA
Bacterial phyla percentages in Terceira and Pico islands
PROTEOBACTERIA
ACIDOBACTERIA
ACTINOBACTERIA
II EIBS, Aveiro 2nd September 2011
Results
• 15 bacterial phyla out of 37 have been found in Azorean lava
tubes
• Acidobacteria (15,3%), followed by Actinobacteria (14%) and
Alpha and Gamma Proteobacteria (12% each) are the most
represented phyla
• 5 phyla, Acidobacteria, Actinobacteria, Proteobacteria,
Nitrospira and Gemmatimonadetes, were found in every
cave
• 12 phyla found in Pico Island, Clamidia, Verrucomicrobia and
Planctomycetes were not recovered. 13 phyla found in
Terceira Island Ktedonobacteria and TM7 were not recovered
II EIBS, Aveiro 2nd September 2011
Comparison of Bacterial phyla recovered in Terceira and Hawai’i
II EIBS, Aveiro 2nd September 2011
Hawai’i
Terceira
Results • Basaltic lava caves from different climate
conditions, one tropical (Hawai‘i) and one temperate (the Azores) share most phyla into their diverse microbial communities
• OD1 phylum was only found in Hawaiian caves
while Cyanobacteria was only found in Azores
• The largest percentage of sequences from the
Hawaiian communities were identified as
Actinobacteria, while Acidobacteria were most
numerous in the Azorean communities
II EIBS, Aveiro 2nd September 2011
On The Importance Of Studying Microorganisms In Caves
Cave Bacterias from Gruta dos Montanheiros, Pico Island
II EIBS, Aveiro 2nd September 2011
Antibiotics
© Airidas Dapkevicius Photography © Airidas Dapkevicius Photography
No antibiotic production
antibiotic production
Pathogens Pathogens
Results
• A seedbank of isolates from azorean lava tubes
was estabilished and can in the future reveal a
valuable scientificly and economically for the
region.
• Lava tubes from Terceira and Pico Island
present biotechnological potential that should
be explored.
II EIBS, Aveiro 2nd September 2011