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throughout history

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hydrothermal systems in the natural

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world have often surprised scientists

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in unprecedented chemistries and biology

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for example the discovery and study of

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the grand prismatic hot spring

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or the lost city alkaline hydrothermal

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vents

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pushed our understanding of the limits

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of life

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and reshaped theories on how life may

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have emerged

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and whether it might exist outside our

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planet

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this is one reason why hydrothermal

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systems are key targets

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in the field of master biology much of

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our understanding of how life may thrive

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outside the ideal conditions in our

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planet

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comes from our knowledge on the

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versatility of microorganisms

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as almost all known organisms that exist

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at the boundary conditions

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or what we may call poly extremophiles

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this exclusivity is tied in with the

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fact

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that some of the microbes that exist

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today

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actually resemble what early life may

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have looked like

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and behaved like press the extremophiles

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life used to thrive in very different

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and much harsher conditions

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one example of such microbial present

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day

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analog are the microaerophilic iron

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oxidizers

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these are proposed to resemble ancient

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microbes

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that may have contributed to iron

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depositions

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in the early earth producing some of the

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banded iron formations

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observed in the geological record

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over the last decade this mechanism of

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iron oxidation

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and the diversity and distribution of

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its microbial agents

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have gained much interest in research

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historically marine iron oxidizing

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bacteria

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have first been described in iron-rich

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shallow hydrothermal systems

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shallow sea hydrothermal vents

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are hot spots for microbial taxonomic

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and metabolic diversity

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they are submerged vents just like the

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more popular

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deep sea hydrothermal vents but because

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of the lower pressure

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there's high discharge of gas

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this process of gas exsolution or

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degassing

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often leads to acidic fluids and enhance

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leaching of metals such as iron and

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this characteristic is true for our

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study site

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which is a 15 meter deep carbon dioxide

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gas rich

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shallow hydrothermal vent in mabini

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batangas philippines

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we observed acidic fluids reaching

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temperatures

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up to 87.5 degrees celsius

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we have also observed high iron

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concentrations

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and low dissolved oxygen ideal

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conditions

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oxidation

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as extension of our marker gene

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sequencing

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for the survey of microbial diversity we

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used an annotation tool

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called fabri-tax or functional

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annotation of prokaryotic dexa

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this is a diagram produced using

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co-inertia analysis

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that fits two ordination plots together

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to

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measure correlation between two

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variables

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what we essentially see is that the

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clustering from the functional profiles

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fit well with clustering from

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geochemical data

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vent specific variables represent the

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upper and lower left quadrants

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along with functions more commonly

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observed in extremophiles

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such as methanotrophy methylotrophy

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and hydrogen oxidation this

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also shows that fibrotax can be an

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effective tool

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in analyzing sequence data sets together

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with geochemistry

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albeit it is only based on pure culture

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studies

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with this limitation in mind we show

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here that it can be used for data

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following our interest in microbial iron

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oxidation

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we were able to detect zeta

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proteobacteria

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in all of our vent samples this novel

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class of proteobacteria

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that are known for microbial iron

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oxidation

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to recap we were able to detect zeta

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proteobacteria

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in the same samples where we've detected

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a wide range of predicted metabolism

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including those typical in other extreme

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environments

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we were then curious of the potential

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role of zeta proteobacteria in our vet

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mats

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no cellulose

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and scattered individual cells within

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the mat matrix

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as visualized in our clsm images

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this characteristic along with the

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filamentous structure

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that were not affected by either

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cellulose or dna strain

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is consisted with microscopic images of

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math forming iron oxide stocks

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of microbial iron oxidizers like zeta

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proteobacteria

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these stocks or filaments have been

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described to take advantage

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of maintained microaerophilic conditions

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we also had the opportunity to look

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further into our 16s

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gene markers against data using zeta

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hunter

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a tool that allows us to resolve zeta

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protobacteria in our samples

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to set out to use from a curated

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database

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from this we detected zeta audios that

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match

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six previously described data

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proteobacterial odus

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namely zeta otu 1 3

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4 21 36 and 59

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two of these have cultural

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representatives described

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in other studies zeta audio 3

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represented by two strains isolated from

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low iron sediments

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and zeta odu36 represented by the

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maripofunda strain

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ekf m39 isolated from the luigi

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obtaining these data otu abundances also

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allowed us to analyze zeta

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proteobacteria with our geochemical data

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by fitting the ordination of these two

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datasets

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we see clustering event samples to the

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upper and lower left quadrants

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represented by both fence specific

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variables which is high

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iron arsenic and organic carbon

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from our data we would like to highlight

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the potential for discovering

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new zeta proteobacteria taxa moreover

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how their abundance strongly coincides

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with our geochemical data

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might inform future hypotheses on the

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drivers of the diversity

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distribution and biogeochemical

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importance

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of microbial iron oxidizers from the

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earth's

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to share the future direction of our

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research

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here is some data on fatty acids from

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the viable components of our

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rust-colored matte

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samples the lengths are more typically

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observed in bacteria and literature

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and a certain compound monounsaturated

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20 carbon fatty acid

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is more often seen in hydrothermal

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microbial streamers than algal mats

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few literature data are available in the

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fatty acid composition of biogenic iron

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oxide mats

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and in zeta proteobacteria cultures

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we anticipate that our data will be

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useful in tandem with our future efforts

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in isolating and characterizing iron

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oxidizers and other microbes

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this presentation is part of a bigger

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paper

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on the microbiome and biogeochemistry of

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the mabini shallow hydrothermal vent

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that we have since submitted and is

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currently being

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this research would not be possible with

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the help of the following individuals

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and organizations we think planet dive

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resort of mabini

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for their assistance we also thank jason

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valaibo

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chica domingo for joining us in sample

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and data collection

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via scuba diving thermo fisher

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scientific and brownson asia's tech for

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technical and logistic assistance

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and this research was supported by the

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department of science and technology

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university of the philippines and chad