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[Music]

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so hi I'm Zoe havlina I am a PhD

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candidate from New Mexico Tech

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um and I would like to just jump right

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in and introduce my field site for this

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project

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which are two cave systems located in

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central Italy so pictured here they're

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very pretty

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and what I can hopefully show you

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um I'm going to take us on a quick field

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trip there is why these are potentially

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very compelling sites for astrobiology

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or for geomicrobiologists such as myself

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and so these caves form through a

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somewhat rare mechanism which is known

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as sulfuric acid speliogenesis so this

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is a cave building process that's seen

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only in a handful of caves worldwide and

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only only a fraction of those can we

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actually access those caves and see this

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process actively happening today and so

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that's what I'm showing you here so

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these are all images from one of my

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field sites which is a Versace cave

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system

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and what you're looking at in this image

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here on the left this is a stream this

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is the groundwater level of the cave and

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it's enriched in hydrogen sulfide and

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this hydrogen sulfide degases where

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there's voids in the host Rock of the

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cave and this hydrogen sulfide fuels

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very a robust microbial communities that

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are thriving off of the energy in this

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otherwise very leukotrophic environment

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and this is seen kind of conspicuously

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in the forms of these charismatically

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named snodite biofilms which proliferate

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in these areas of the cave here's

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another picture of what they look like

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and we also sometimes see if you can see

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the yellow dots native sulfur that can

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precipitate in these areas as well

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and these are primarily composed of

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citythial bacillus which is a sulfur

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oxidizing bacteria but what I'm

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interested in for the context of my

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project are all of these white I don't

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know if you can see my pointer here

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these white clumpy minerals here and

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this is actually microcrystalline gypsum

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this precipitates in part kind of as a

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consequence of this microbial activity

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because the sulfur oxidizers are

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producing sulfuric acid which is

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obviously highly corrosive when you have

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a carbonate host Rock and so this gypsum

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is precipitating as a replacement as

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this host Rockets corroded

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and it's highly acidic so pH is usually

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zero to one and what happens is that

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this gypsum can build up in these cave

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systems and remain there essentially as

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long as you don't have any Waters that

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come in and wash it away and so that's

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what I'm showing on this image on the

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far right here this is one of these like

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massive gypsum deposits that exists in a

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part of the Versace cave that hasn't

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been exposed to this active sulfide

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degassing for maybe a hundred to two

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hundred thousand years

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and so we see sulfates and gypsum on

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other locations notably the surface of

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Mars so I'm showing you data here as it

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relates to the Gale crater

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and there's some evidence or some

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thinking that this sulfates could have

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also precipitated under acidic

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conditions so potentially these are

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thought of as

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places where we might look for for

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evidence of life and on the Earth's

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surface the analogs that we have there

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are several and these have been well

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studied by other teams of

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astrobiologists but what is novel um or

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the argument I'm making for these caves

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is that this is an ecosystem that as I

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just showed you is fueled by chemo litho

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autotrophy so these kind of alternative

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Lifestyles and so potentially studying

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the gypsum and its ability to preserve

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or not preserve

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biosignatures is is kind of a novel

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aspect and gets away from surface world

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where we sometimes see the signatures

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mostly of things that are photosynthetic

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and so in the context of this talk when

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I'm talking about life detection or I'm

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talking about signatures I'm talking

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about lipid biomarkers specifically and

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so what I'm showing you here are some

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types of these they were kind of

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explained earlier these ones on the left

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which are those hopenoids so these are

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produced by bacteria and these can be

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functionalized and then they have this

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very recalcitrant

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backbone structure this hoping that is

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this diagenetic structure that can

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persist in some terrestrial environments

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for potentially millions of years and

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then on the right these are

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gdgts from archaea so kind of a similar

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lipid biomarker and the these specific

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structures that I'm showing you are ones

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that have been classified from those

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snodite biofilms that I showed you in

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their earlier image

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and so knowing what we already know

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about these cave systems which is that

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we have gypsum that is precipitating in

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in very close association with microbial

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activity that's kind of these microbes

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that are using an alternative lifestyle

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and we know that some of these organisms

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like that aciditythyl bacillus are

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capable of producing these lipid

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biomarkers and this gypsum is kicking

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around in this cave for a little bit of

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time we sort of used what we know to

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form these questions about things that

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we don't know

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um we don't actually know or going into

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the study no one had ever looked at what

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microbes are colonizing the surface of

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the gypsum itself

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and we don't know if those organisms

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produce their own lipid biomarker Suite

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if we'll be able to characterize that or

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detect it in the gypsum at all and then

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if we do does that persist in this

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gypsum that has been existing in this

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cave for a longer period of time

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and so I won't go into the Weeds on

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these methods here but I just want to

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highlight that we're using kind of a few

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different approaches to answer some of

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these questions so using some sort of

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classic molecular biology techniques

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amplicon sequencing and metagenomics to

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characterize those microbial communities

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and then using some organic geochemical

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techniques to try to characterize those

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lipid biomarkers

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and so some of the data that I've had

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um is that these gypsum surfaces are in

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fact colonized by microbes and so what

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I'm showing you here is amplicon

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sequencing data so that's 16sr in our

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sequencing that tells us who's there

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and what I want you to take away from

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this graph is that I'm plotting two

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things here on the left to this access

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and the blue dots is showing the percent

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of that acid ethyl bacillus so that's

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that main sulfur at oxidizing bacteria

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that we see in those snodite biofilms

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and then on the right I'm showing in

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this orange I'm showing you the overall

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diversity of these samples and so all of

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these dots are gypsum samples that we're

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taking at kind of varying distances from

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that active lead degassing sulfate

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extreme

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and just a reminder so these biofilms

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that have been previously characterized

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are around 70 percent of citythal

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bacillus and so the main takeaway is

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that are samples of gypsum are far more

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diverse than that you know even at this

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kind of like best case scenario where

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we're the closest to the sulfate extreme

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we're really not seeing anywhere near 70

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percent of city-thal bacillus and so

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this is data from that other cave that

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I've studied and I don't have that nice

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distance gradient but again it's

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plotting the same things and I've just

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kind of broadly lumped these samples

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together in terms of like high level of

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sulfide medium level lower levels and

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again we see a similar Trend we also see

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low levels of acetyl bacillus and in

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fact we see a heck of a lot of archaea

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too so some of these samples were almost

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entirely archaea

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and why I talk about acetyl bacillus and

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I talk about the diversity of these

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samples is that you know we know acetyl

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bacillus is capable of producing lipid

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biomarkers but it's one of the bacteria

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that can and in fact a lot of them can't

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and so this potentially has implications

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when we think about you know are there

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things that is in this gypsum that is

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producing lipid biomarkers or aren't

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there

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and we can kind of get at this um

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indirectly through metagenomics and so

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again metagenomics is looking at all of

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the genes that are present in a

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particular environmental sample and so

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far I have only reconstructed a single

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metagenome and this is from a native

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sulfur deposit but I did do some Gene

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searching and I was looking for this

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squaly and hoping cyclase Gene which

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forms a critical precursor to making

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those hopenoid lipid biomarkers and I

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did find some copies of that but again I

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also found a ton of archaea and two of

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the high quality bins that I was able to

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assemble were these two kind of Novel

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archaea that have potentially an

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interesting symbiotic relationship and I

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would love to talk about that more but

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that's kind of a whole other talk so I'm

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just going to leave it at that with this

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complicated figure here on the right

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um

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so it you might be guessing or kind of

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wondering why I've only gotten one

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metagenome so far and really it's

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because the overall biomass in these

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gypsum samples is very low and so what

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I'm showing you in this picture here so

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this is microscopy this is dappy

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staining so looking at cells and so you

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see hopefully you can see just a few

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blue dots those are cells and on the

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right this is showing them sitting in a

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in a bathtub full of gypsum micro

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crystals and so when you just kind of

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contextualize this low number of cells

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in in the vast array of all of these

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minerals

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there's not a whole lot going on and in

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fact some of our preliminary total

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organic carbon measurements are less

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than one percent or even a fraction of a

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percent and so

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that all leads to kind of some

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complications with trying to actually

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extract some of these lipid biomarkers

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and so this side of the project is still

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an active protocol development we

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haven't seen anything yet

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um and so we've had to kind of scale up

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our extraction we're hoping to extract a

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lot more gypsum and and I'm hoping that

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that's something I'll be able to talk

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about more in the future but I just want

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to return to sort of The Guiding

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question of this of this study which is

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you know looking at these

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very

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um interesting microbial Dynamics the

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way that the stripsum is formed we see a

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lot of we see biofilms existing where

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where the gypsum is forming but we don't

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know

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if there's biomarkers that are preserved

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or not and I have to leave that question

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open at this time

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um but hopefully I can answer it in the

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future

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so I think I will just leave off there

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and

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um I wouldn't be able to do this work

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without I have a lot of very fantastic

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collaborators and funding sources and I

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would especially like to thank all the

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folks at the astrobiology analytical

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laboratory at Goddard and I think that I

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have some time for questions so thank

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foreign

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hey I'm bunny Chase I'm from the jet

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propulsion laboratory I really love this

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question you're asking and like the way

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that you're trying to attack it and I

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was wondering

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a lot of the lipid biomarkers that you

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talked about looking for they as you

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said can be preserved for a few millions

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of years but what about the more stable

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versions like looking for her pains

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rather than hope Knowles are you also

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looking for those yeah yeah we are I

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didn't really

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explain that super well but we are

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hoping to look for helping hoping to

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look for Hope pains as well yeah

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um what do you think like the Pres

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the precious in this cave would be for

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the molecules and how similar is that to

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Mars yeah I think that that is kind of

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the key question for sure

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I don't know if that's something that I

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want to dive into right now but I think

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we should talk about it later for sure

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I'm curious to hear your thoughts on it

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as well

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um obviously this is a oxidizing

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environment it's it's acidic it's wet so

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it has a lot that could maybe not be

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preserving things but

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um you know certainly folks have done

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similar research and and have found

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biomarkers in in Gypsum and other

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environments on Earth so

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any last questions for Zoe

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okay thank you so much oh wait there's

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hey so I have a question about the

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microcrystalline nature of that gypsum

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so what do you think that might what's

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your hypothesis for structurally how

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that might affect preservation of Bio

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signatures as opposed to other kinds of

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gypsum that might happen on the surface

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yeah I think that's a really question

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um

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I guess you know off the top of my head

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I would say that

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I don't know if we're gonna get it like

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doesn't really layer I think necessarily

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in the way that we see in like

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evaporated gypsum

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um how that may or may not you know

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factor into biosignature preservation I

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don't I can't really speak to that but

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it is I think it's definitely a really

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interesting difference

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um that we see in the strips on for sure

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and it's like

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it's I didn't really show any of the

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like there's that one picture that I

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showed of the gypsum but it's it's crazy

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it's like you know we call it Rock in

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name only

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um it's it's like goo

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um you go to sample it and it's almost

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like a non-Newtonian fluid it like just

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kind of like oozes

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um which is super wild and and it's kind

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of difficult to work with to be honest