Hello my name is Blake Lindgren
and I'm presenting Finding
Caenorhabditis Nematodes in the under
Explored Midwest
So the first thing you need to know
What C Elegans is and what the project
goals are C elegans is a free living
Bactriovorus Nematode it lives in
Decaying matter and is found across the world
And is a powerful genetic model organism
The project goals are obviously identify
and find a source of C Elegans
Contribute to C elegans biogeographic database
and characterize isolation sites for
C elegans
So you know we're looking for C elegans
and how do we get them is
we isolate them which means we take
Decaying matter which is rotting apples
Rotten walnuts or compost or other things
And we try to put them on plates and pick
the worms off to keep them
we have isolated previously
Caenorhabditis sp. 8 and C briggsaes
C elegans has not been isolated yet in
the Midwest or in Nebraska
This is the map of sites we are going to
isolate or to get worms from where we
going to sample from martins hillside
orchard valas pumpkin patch and east side
Campus from the university of Nebraska
Which you can see here on the right side
so the first place we went to was east side
Campus and we were sampling the walnuts
From there and we got three walnuts from
the greenhouses over here and we got 22
samples from the beehives over here on
east campus we can see in these pictures
the next place we went to was martins
and Valas and I forgot to mention we did
not get any nematodes form the greenhouses or the beehives
The next place we went to was martins and
when we went to martins we sampled 46 apples
and we only 2 nematodes from a single apple
from this side area over here'
and the next place we went to was valas
we sampled 31 apples and got zero nematodes
from them as well
and I should mention about the apples
very quickly is this is not normal
last year when they went to martins
they gotten all there nematodes form one sampling
and they didnt sample 90 apples they
sampled the same amount of apples so this
is very strange to say the least while we haven't
sampled walnuts as I mentioned before this
is far stranger because we have no reason
to believe normal this has not happened before
in the research years so its defiuntily
a bit strange and something to keep note of
so after that we only have 2 worms
we need to move on and find something else
so next thing we try to get is compost sampling
and we got our compost SBS faculty and students
And they got us 18 bags total
out of the 18 bags 16 had worms and out
of those 16 two different kinds of worms in them
so when we sampled the bag we had two
different district kind of worm's
and this is the result tab of them and
you can see we sampled alot but we didnt
actually get that many compared to last year
we definitely had to sample a lot more this year
but 16 of the worm's came from compost
2 of the worms came from apples
and once again to mention this is not
normal for apples apples should have
given us a lot more compared to what we have
gotten in years prior
so we're kind of gonna go through the
workflow really quick the first thing we
have to do is we have to do is isolate
our nematodes which we just did we have
gotten our nematodes we got them on our
plates and what's the next step and how do
you tell worms apart so really your got to look
for is the growth rate and the self fertility
and we check those rates and ill explain them
in a sec but after that you look at morphology
and then you do a DNA sequencing of a gene
now for the self futility and growth rate
to briefly explain growth rates are
comparative that's the best way to describe
them but we do look at some other things
we look at basically how fast they grow compared
to other ones and how fast they grow
compared to out C elegans that we have in
lab because we do have some C elegans
from other parts of the world
but how fast do they just comparatively
Because that's the best way to compare growth
rates very easily at least in a short amount of time
so fertility is also very important Because
C elegans is a hermaphroditic worm
Which means its self fertile it means it
only needs itself to produce offspring
and C elegans is a fast grower
so you do that very simply by picking a
female to a plate and waiting for it to
reach maturity and then once it reaches maturity
if it has offspring its self fertile
because its by itself and it had to
fertilize its own eggs
So the next thing we look at is morphology
and we really look at two big traits this
can tell you alot about your worms the
two big traits are buccal cavity and the
pharynx you can see these two diagrams
here and we check under a very powerful
microspore to look at the buccal cavity
and the pharynx ill show you better on a
worm that I have so the left picture here is
the buccal cavity and the pharynx is the
right picture you can see the mouth structure
its a very distinct feature and its a lot
easier to tell apart then other features
which is why we use this two predominantly
so after you've gotten your worms and you've
done fertility' tests and everything you
have to ask what worms are you gonna
sequence and which ones do you want to do first
and which ones look the best
so what your looking to do is
tell your worms apart you cant do the
whole DNA strain because that would be
ridiculously long and a pain to deal with
so instead a much easier way to do this
is to take a conserved gene which is a
specific kind of gene that basically
allows you to easily compare your worms
against each other because it is conserved
this is a important part because it basically
means there are very few changes and the
changes that happen would be very specify
to species so we do this by taking the
18srRNA gene which once again a very specific
gene all you really need to know is that
it is conserved and it allows us to compare
worms very easily and we do this by doing
PCR and then gel extraction
the results from that are here from one
worm just for showing you
on this example we have the results at the
bottom here over at the qaury cover
that is the percentage of likely hood
it is that from the DNA we have its 99%
for the top 2 and the top two are
Rabdiltella axie which is a good likely hood
that actually what it is and ill show
you quickly were that is on the graph here
now this red line here is the
specific species and the bottom is C elegans
and there distantly related but it is the
result of them and that's how we compare them
moving forward so really the future directions
of this project is obviously characterize
all the other C legans candidates we have
and the ones don't have so we can double check
they are not C elegans or related because
morphology does not always tell you
everything you need to know the other
thing we need to do is investigate the lack
of nematodes from apples that is very strange
from this year I cant explain how strange
that was when we sampled upwards of 90
apples and only get 2 worms from that
and last year they sampled 40 apples and
got plenty of worms the next thing we need
to do after that is to gather more samples
not just from apples but from other things
and see if C elegans is living in those
so this is very important we do those things
and the final thing id like to do is to
acknowledge the people who helped me
so id like to thank all of Herman lab
but really id like to thank Dr Michael Herman
Leah and most importantly Ashley
they all really helped me but Ashley
specifically but they all helped me
and the program id like to thank is FYRE
being a freshman it was most freshman
don't get to do research and id just like
to thank them