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you

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[Music]

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hello I'm Mary Sue Buddha I'm from

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Michigan State University and I'm going

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to be bringing up some topics that have

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been covered a little bit in the

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previous session and some things that

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people have touched upon throughout apps

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icon so far but the point of this talk

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is to look at five geochemical

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interfaces in 17 izing systems at a much

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finer scale specifically we focused on

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the case study in June 2016

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at the coast range a few light microbial

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Observatory so surprising systems we

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know are common throughout the world in

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Oman Portugal Italy Newfoundland and LA

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City but for this work we study at the

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coast range Oh feel light in California

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it was in place 155 270 million years

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ago during the Jurassic period and we

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know that serpentinization mobilizes

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volatile such as hydrogen and methane

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and those ground waters can host reduced

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carbon compounds that microbes can use

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as sources of nutrients and energy

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however because the environment is

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extremely alkaline and there's low

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dissolved oxygen

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it creates really harsh environments for

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microbes to live so it's important to

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study these organisms and understand how

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they can cope with these stresses and so

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dependent associated fluids can also

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lead to limitations and availability the

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availability of nutrients such as di C

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to support growth because calcium and

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carbonate can react to precipitate s

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calcite so chromel is about three hours

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north of San Francisco of Lex my

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colleagues mentioned earlier look there

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are two well sites well clusters Cory

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Valley no.8 to the west and then core

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shed Wells to the east they're about one

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and a half kilometers apart they are

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drilled in 2011 and they are drilled to

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a bunch of different depths in order to

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gain a big-picture perspective of the

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subsurface at crumble and kind of gain

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insight into processes happening with

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depth so as a result the pH ranges from

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seven and a half to twelve dissolved

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oxygen ranges from point zero nine to

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two point four

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and eh or oxidation reduction potential

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ranges from negative 297 which is very

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reducing to 116 millivolts which is

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pretty exciting so the point of our

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study was to because serpentinization

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leads to natural gradients in water

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chemistry and all life as we know it

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requires generating energy from chemical

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gradients so we wanted to elucidate the

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relationship between microbial community

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compositions and chemical variances

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within the serpentine high ground water

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and then use that as an analogue for

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guiding our efforts in the search for a

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stir biologically relevant surprising

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systems in life elsewhere so what we did

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is in June 2016 we created a top-down

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profile of one of the well c SW 1.1 and

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the purpose was to understand how those

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chemical gradients within the energy

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limited and serpent event hosted ground

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waters could influence microbial

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community compositions and I just want

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to emphasize that this is one of the

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first comprehensive Wells comprehensive

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profiles of us repent o night hosted

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groundwater well so it's pretty

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important but this is the schematic I

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came up with for this I want to explain

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it in detail a little bit but we thought

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really long and hard about how we would

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create a profile without disturbing the

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water column and within the time

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constraints we had available to us

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that's what we did is we took tubing the

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depth to water meter an ultra-sensitive

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dissolved oxygen probe and sterile tygon

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tubing zip-tied together ethanol

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everything made sure was as clean and

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sterile as possible and then hooked one

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end of the tubing up to a peristaltic

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pump at the ground surface and then

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lowered the collective tubes into the

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well proof the top of the well in order

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to start sampling and so we chose for

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dissolved oxygen depths as a start so we

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chose 100 percent air saturation 50

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percent 15 and 0 so as we went down we

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saw

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but when we normally pump the well at

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the bottom of the well it's extremely

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anoxic so we realize there'd be a

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gradient of course with that and so we

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used it as a proxy

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so we nos are cations anions organic

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acids gases sulfide BSE and your water

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quality parameters as well as cell

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abundances cubit for DNA fluorescence

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and 16s RNA sequencing so the idea is

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after we finished pumping from the

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previous trip the water would infiltrate

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riah collaborate with the groundwater

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with the new conditions and then the

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gradients between the surface or air

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saturation and the bottom of the well

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would develop and things would stabilize

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and then during that process microbes

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could take advantage of the gradients

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and the mic for basically generating

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energy so what we see is other than

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dissolved oxygen decreasing pH

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temperature pH and conductivity were all

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analyzed and it looks like between 9.2

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and 4.6 pH decreased but then

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temperature eh and conductivity

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increased and then between four point

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six six and one point four pH increased

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and then subsequently temperature eh and

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conductivity decreased so we see some

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interfaces going on there and we want to

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explore this a little more through

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microbial analysis before you do that we

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analyzed a bunch of anions and dissolved

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gases so in the top right here we have

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hydrogen and carbon monoxide all these

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plots show dissolved oxygen on the y

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axis and then concentration of the

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parameters are on all the x axis x axis

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but hydrogen and carbon monoxide up here

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generally show an increase with

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decreasing dissolved oxygen v.i.c and

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methane show a decrease bromide of

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fluoride show a decrease and then

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hydrogen sulfide and sulfate redox

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parameters show opposite trends which

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indicate that

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at with decreasing dissolved oxygen it's

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becoming more reducing what's up so and

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then chloride also shows a decreasing

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turns so we'll draw preliminary data and

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we need to look at the cations and

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anions and balance when we get that data

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back to really understand what's

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happening but generally in terms of cell

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abundances microbial cell counts a

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decrease in cell abundance was observed

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as oxygen decreased in the wealth as

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so from 16s sequences that we received

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this one here as the 4.6 six milligrams

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per liter dissolved oxygen and then over

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here is the 0.075 or deeper into that

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profile and always see as a dominance of

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beta Proteobacteria as we've heard quite

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a lot throughout these talks they

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dominate and interestingly we see more

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diversity in that bottom of the well

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they're like the 0.075 level and that's

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likely because see some1 one is case to

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a depth and then it becomes uncased and

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so it's likely that we pulled some

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laterally influenced groundwater from

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that area but we also see Dino cacti and

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then Rico micro BA and cluster dia as

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well dominating so the implications for

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this are that we likely see some

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recharge happening in a well that led to

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subsampling sub lateral groundwater flow

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but the cluster D on the route through

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Co microbial and all their smaller less

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dominant species in these areas should

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be looked into a little bit more in

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depth to see who's there and exactly

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what they're doing but the idea is that

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those other organisms may be thriving

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off the reduced products of

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serpentinization so additionally after

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looking at this profile we compared it

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to the bottom of the well where we

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typically sample and what we see is that

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basically sulfide increases and sulfate

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decreases with decreasing dissolved

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oxygen and then pH increases and

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conductivity also increases so it's

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getting a little bit higher in

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alkalinity and a little bit saltier so

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when we compare the 16s sequences from

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our two analyses to the well bottom work

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by Katrina Queen and authors in 2017 we

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see that it's mostly beta bacteria and

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Firmicutes again but the pH is really

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high it's still 12

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so it's likely that serpentina bonus and

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which is another beta beta

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proteobacteria is dominating there

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because it's tightly coupled to pH and

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pH 12 specifically so another figure by

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Katrina tweak this is a PCA plot just

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showing that generally chromel organisms

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are influenced by a range of chemical

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parameters for instance QV one one is

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appeared by depth and temperature

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CSM b13 is influenced by temperature and

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ORP

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but what we're looking interested in SES

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f/11 and they are influenced by organic

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acids predominantly pH and conductivity

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so when we get to organic acids added

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back we can see at the top of the well

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is still in line with what we see at the

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bottom of the well but yes then we

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compared the chromo data to the season

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one one data to the other wells at

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chromo and we see that beta

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proteobacteria alpha and gamma dominate

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in QV 1 - beta proteobacteria

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austria and alpha dominates in CS 3 1/3

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and then beta clostridia pastila and

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gamma proteobacteria dominate throughout

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QV 1 1 and so it's likely that there is

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a more bit more diversity here because

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it's less alkaline and the conditions

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aren't as harsh for organisms to survive

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even if just by a little bit it may be

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just enough so just to conclude we still

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need to get 16s sequences back from the

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very top of the profile and the 15%

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interval so when we do that we'll be

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able to get a full column geochemistry

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and microbiology didn't really

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understand what's happening but perhaps

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we can also

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create profiles of other Wells in the

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system to compare and analyze and see if

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this is just happening in CS everyone

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one or if it's consistent throughout the

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rest and then this is important because

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that Union it lends unique insight into

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the variability of microbial species

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that can inhabit serpentine izing

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systems and the extreme conditions and

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then this will help us understand the

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distribution and activities of microbes

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in the subsurface at chromel in order to

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help us detect life in other aster

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biologically relevant scenarios so with

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that I would like to thank that Nai

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early career collaboration award because

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without that this would not have been

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possible and also Torrey and Mike for

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hosting me at Ames this past summer it

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was they were a huge help and then the

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shrink lab at Michigan State and the

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rock powered life team we're also key in

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this analysis so thank you alright we

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have time for one question and if the

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great talk Mary thank you I noticed that

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you had a spike in your geochemistry as

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you are going down the the do gradient

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so not at the very bottom but the one

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right above it do you have any idea what

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the geochemical or microbiological

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reasons or implications could be of that

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so I think it's partially because we

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have the real collaboration of the well

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but then also when we were pumping we

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saw a little bit of recharge Japanese so

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the water actually increased a little

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bit so I think we are pulling laterally

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from some probably some surprising water