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[Music]

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in this talk I'm gonna present about the

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early findings of a collaborative

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research done between my department the

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Department of Microbiology analysis and

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the department of geological science

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both of us working in the university

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Florida so what system are we working

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we're working a system called the upper

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florian aquifer and the upper florian

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aquifer or UFA for shirt is yes an

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aquifer so now then the next question

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what

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hydrological formation that contains

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water within it pretty much like a

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sponge in the case of the you FLE this

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aquifer is a karstic aquifer and by

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karthik i mean that the rock that

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contains the water is limestone and in

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fact limestone formations as you can see

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in this map are widespread across the

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globe they cover around 20% of Earth's

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surface

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and in fact in the case of florida they

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the ufa extend pretty much through the

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whole peninsulas well some parts of

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South Georgia and Alabama now the deal

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here is that this aquifer is what's

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called a confined aquifer meaning that

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there is a layer of rock on top of the

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rock that contains water that is

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impermeable doesn't allow the water to

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go through whoever in some parts namely

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North Central Florida this top confining

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unit has been removed by erosion and -

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and that's not a surprise that all the

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six prints though we have simple are

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located in that uncle finery now you

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might be asking yourself okay you have

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this cool system but why is it relevant

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for us to biology because we are in an

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astrobiology conference so there are

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many points that I can talk to you about

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this aquifer and whites rallying for

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astrobiology here are the main ones

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first of all this aquifer is limited in

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any surface inputs there

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one of the aquifers that we believe

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they're completely shielded from input

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from the surface meaning that these

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specific points are low energy systems

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and by that all the microorganisms are

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living here should adapt to deal with

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these low nutrient concentrations also

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we have recently discovered that these

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aquifers owners aquifers as well are a

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source of poorly understood microbes

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that can span over bill the Tree of Life

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and finally very cool discovery as well

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that we have discovered that both encho

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and methane both green houses are

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emitted from this system now when we

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talk about your chemical parameters

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within the UFA in general we will find

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that some geochemical parameters like

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what's the temperature salinity pH they

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do not change at all no matter the

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spring we go no matter the time of the

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year we go there these guys remain

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constant so these parameters are not the

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ones that are going to form us about

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processes that are happening in

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different part of the UFA whoever we

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have all parameters namely actual

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concentration in the water or oxygen

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concentration that do change either over

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time or from spring to spring now it's

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worth note that oxygen concentration

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versus entry or concentration Falls

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positive correlation except there are a

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couple of exceptions there but those are

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very particular places and this positive

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correlation with oxygen is make a star

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making us thinking about processes that

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might be producing this ain't sure

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whether there's one particular location

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called the river sink on rice and in

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this particular location we have a river

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that sinks under water underground sorry

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mixes with the UFA water and after

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around 7 kilometers it rises up again

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so in this rice after mixing with that

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UFA water

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well we have found it and

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shuo concentrations not only changed

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over time but that since also fools a

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negative correlation with oxen now we

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reach the question part how this angel

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is produced in the system well I wish

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that I could answer this question right

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now but we don't know their answer yet

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whoever we believe that the main

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processes responsible for entry or

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production in the whole nitrogen cycle

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could be responsible for Angel

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production in the system as well

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what are those processes first of all we

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have nitrification

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nitrification is the oxidation of

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ammonia all the way to nitrate and this

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process requires oxygen to happen in

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contrast we have the nitrification which

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is the reduction of nitrate or the way

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to NGO although some microorganisms

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actually stop at the level of n - n - o

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instead of the level of n chew so this

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gas is released as a consequence and the

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nitrification is anoxic it is inhibited

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in the presence of oxygen and finally

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there is a third process that is micro

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Felix or requires just a bit of oxygen

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to happen and it could be seen a circle

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as a combination of both processes

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called nitro fire denitrification so do

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we have now any hints that beyond that

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correlation with oxygen that I'll show

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you before that coming from about if any

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of this process is happening in the

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aquifer yes we do we have 16s ribosomal

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RNA gene data and with that data it's

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difficult to tell any specific pathways

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are present but at least we know that

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some micron is microorganisms that do a

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specific pathways like this file a time

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where Keota belonging to the mine

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archaea these guys are known to be emoni

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oxidizers which is the first steps of

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nitrification and as you can see they

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are presenting

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no but still significant amounts in some

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of the springs so this might be pointing

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out to me tooth decay should be a

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possible way to put this ancient or at

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least in some of these locations in fact

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I just want to stress that this LRS is

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one of the places that the most angel

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has been detected

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also because we still don't have

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metagenomic data we use program to pie

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crusts to predict some pathways that

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will be present in this aquifer is based

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on the 16s data and while we found

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interesting findings here is that the

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nitrification is relatively abundant in

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the lena river rice which was that

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specific system that i mentioned you

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before so this further strength this

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correlation with this negative

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correlation with option that we saw

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before and another thing is that this

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problem point choices that

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methanogenesis although in low numbers

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seems to be present in some of the

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aquifers well whoever one we met when we

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measure methane in all the springs we

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did not detect any methane above

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atmospheric concentrations except in one

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place there's that particular location

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that I mentioned to you before this

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river sinking rise in this specific

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location methane follows an interesting

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trend this is the methane concentration

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the water and the delta 13 values for

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the water is going in and you can see

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that these values are relatively

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constant whoever in the water is going

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out

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they both methane concentrations and

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delta 13 values changes a lot so we

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believe this could be correlated with

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both nutrient inputs oxygen the changes

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in oxygen concentration as well as yes

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amount of water is going into the system

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as well whether that doesn't mean that

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methanogenesis is not happening in any

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parts of the aquifer because according

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to verse 16 is theta we have detected

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Europe Yatta which most of the nervous

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and we also have detected methane odors

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being methanogens the guys that produce

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methane and methanol drops the guys are

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consumed methane oxidizing it well there

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is a important thing here to note that

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as you can see methanogenesis is a

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strict anaerobic process and it's

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inhibited if oxygen is present whereas

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methanol trophy is a microbe philic

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process requires a bit at least of

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oxygen to happen now if you look at the

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data here

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well there's places that only

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methanogenesis are present but there are

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all places where we can find both

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methanol drops and methanogens methanol

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ian's together now remember that we were

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well we are simply where we are sampling

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here is just a water is going out from a

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specific spring that water is going to

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collect different microbes from this

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different place of the sea of the system

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so these guys definitely cannot live

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together but the father we detect them

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in our samples means two things that

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methanogenesis could happen that that

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methane is probably consumed because we

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don't detect that in our water samples

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before it relates to the surface and

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also that this system that we are

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working is algae rings so an important

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thing

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another cool finding is that we have

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detected these microorganisms that

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belongs to these poorly known groups

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only one is a bacterial group also known

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as bacteria the peninsular peer group

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that encompass different archaea phyla

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although they belong to these two

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different domains of life they share a

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lot of similarities they are they have

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reduced genomes they are believed to be

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some by oats they are anaerobes they are

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believed to be fermenters and they are

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very small and you may ask yourself what

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you're talking about bacteria of course

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they are going to be small well these

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guys are small even for any

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microorganism they are small enough the

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go through zero point

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to micrometer filters which are in

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microbiology they are assumed to retain

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pretty much all microorganisms well

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that's why that's the reason we use 0.1

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filters to sample the water and we have

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detected that in this of 0.1 micrometer

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filters the the populations of

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microorganisms are composed by up to 70%

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of these guys so that that is surface

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appearances in finding and the main

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question is and the reason I brought up

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this microorganism is here in

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relationship with this greenhouse gases

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that we have detected is the fact that

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we don't really know what these guys are

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doing and they might be even carrying

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out pathways that we don't know yet that

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nobody has described it and that might

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contribute to this either methane or

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entry or production so now yes to

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conclude and remark the three main

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points of this talk first of all we have

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detected methane at least in some

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specific locations of decision and encho

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above atmospheric concentrations in all

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the springs that we have simple how

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these gases are produced we don't really

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still have the key yet especially for

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intro but we believe that you know the

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production is a strongly related with

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microbial activity and we would like to

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in future studies see if there are any

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particularity of entry on methane

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related with microbes that made the use

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of these two gases as potential

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biomarkers then of course the aquifers I

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mean as I mentioned to you before seems

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to be a heterogeneous environment and

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that will be a very cool system to

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compare how some microorganisms are able

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to thrive in a low energy environment

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and determine exactly what's the

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contribution of any nutrients from the

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surface for the habitability of

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specific extreme environment and finally

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the father we had detected this bully

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study microorganisms not only opens the

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door to new pathways dare to be

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described but also it allows us to

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expand our view of the Tree of Life and

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what are the actual limits of life not

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only in terms of physiological limits

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but even in term of size limits as well

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so with all of that I would like to

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conclude thanking of course on my

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committee members all the members of my

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lab as well as all the collaborators

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have have made this project possible and

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of course the University of Florida by

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University Institute for providing the

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funding for making this very possible

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with all of that thank you very much for

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your attention and I will be open for

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any questions so as you alluded to

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earlier you haven't you don't know what

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these organisms are doing have you do

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you guys have plans to look at the meta

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transcriptome of the samples that you

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look at so far yeah well one of the next

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step of this project is definitely look

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at metagenomics in general and see not

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only what these guys are doing in the

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aquifer but also what other organisms

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are doing as well as specifically for

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these organisms the members of the

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Defiant phyla

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they are believed to be anaerobes and

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some of them fermenters just based on

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methods in an exercise that other

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authors have done and they some of them

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they lack of many of the machineries for

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production of amino acids and lipids and

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even synthesis of nucleic acids so some

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of the authors believe that they might

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be some buyouts of other organisms and

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if they are sent by H we would like to

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know exactly which organisms they are

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okay if there are no and if they're not

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any other questions Thanks

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could you speak a little bit more to the

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water column chemistry as you know it

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and if there's any significant sort of

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chemo client's going on there so it's

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important to note that we're not working

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in a lake we are working in an aquifer

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so any nation of water come will be a

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bit biased because you know it's not

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that we stick a probe underground you

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know then we have like an ocean layer

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and then on an ox layer or something

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like that it's more like it's more of

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geographical spread of the indecision

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that we have some Springs that all these

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Springs are oxic and that's there's the

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only water we can sample is something

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the water is discharged from these

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Springs weather although in all the

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springs are exceed their some of them

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that have very load you we're I'm

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talking about concentration around 1.5

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to 2 percent D so function and there are

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other Springs that have relatively high

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values the highest and members around

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55% is so function so we cannot talk

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about an actual water column but

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definitely we can see geographical

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difference between the difference

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principle Center thanks are you choosing

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Springs that have a lot of human some of

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these Springs are pretty busy I don't

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know exactly know which ones you're

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using but I know some of the Florida

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Springs get really busy with people and

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are you trying to control at all for any

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human influence well we don't really

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care that much about human influence

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because many of these Springs they have

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like a very strong current in fact

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something there is a challenge because

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it's very difficult just to swim above

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the bend where we have to deploy our

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instruments

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so if there's any humans with their

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bacterias around now everything will be

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washed off by the water current itself

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another thing of working this very

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populated spring specially the summer in

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Florida you know they are even better

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than going to a beach for most part is

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that you know it's that interaction with

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00:18:08,419 --> 00:18:05,909
people that makes easier kind of like

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diffuser where our work many people use

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00:18:12,260 --> 00:18:10,260
concur is like hey what you're doing and

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00:18:14,000 --> 00:18:12,270
we have to pretty much explain in the

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00:18:16,130 --> 00:18:14,010
field what what they are we are we are

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00:18:21,700 --> 00:18:16,140
doing with our wind instruments and all

385
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ok one more quick question so it's got

386
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to be very quick but so that's a very

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populated area as some people alluded to

388
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so I'm actually wondering if you did any

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sort of mapping to see where the

390
00:18:39,940 --> 00:18:37,799
research for the aquifer is and if that

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can influence the amount of nitrates or

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ammonia or nitrogen that's in the system

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already by the time it goes through the

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spring yes for the research of the

395
00:18:50,450 --> 00:18:48,210
aquifer there are two main areas when

396
00:18:52,700 --> 00:18:50,460
it's like a lateral which starts coming

397
00:18:54,889 --> 00:18:52,710
from the north and then there is all

398
00:18:56,570 --> 00:18:54,899
this sunken finally that of course is a

399
00:18:59,210 --> 00:18:56,580
recharge area and there are some

400
00:19:00,769 --> 00:18:59,220
problems here with you know nitrogen

401
00:19:03,470 --> 00:19:00,779
fertilizer runoff

402
00:19:06,230 --> 00:19:03,480
so actually in natural concentration the

403
00:19:07,870 --> 00:19:06,240
waters relatively low but we believe

404
00:19:12,399 --> 00:19:07,880
that some part of the problem is that

405
00:19:14,630 --> 00:19:12,409
that nitrate or some other

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00:19:17,060 --> 00:19:14,640
transformation other species of nitrogen