Jump to content

e-DNA Sampling For Sasquatch


Recommended Posts

Wolfjewel

If anyone else, like me, feels a little out of the loop in understanding environmental DNA, I found a good primer on the topic, with scientific citations:

sciencedirect.com, search box “Environmental DNA – An emerging tool in conservation for monitoring past and present biodiversity”

I had to create an account, but that doesn’t seem like a problem. 

Happy to see the use of the word “cryptid” in the abstract of the article! Sorry, I couldn’t get a hot link to it to share here.

 

I would be glad to get samples of water from the Hockomock Swamp in SE Mass., known as a cryptic hotspot, but sightings are old at this point.
 

Edited by Wolfjewel
  • Upvote 2
Link to post
Share on other sites
hiflier
Posted (edited)
16 minutes ago, Wolfjewel said:

I would be glad to get samples of water from the Hockomock Swamp in SE Mass., known as a cryptic hotspot, but sightings are old at this point.

 

Isn't there kind of a famous name for your area? A "something" Triangle? Seems to me a saw a book somewhere about the strangeness that goes on around there.

 

And while "environmental" DNA sounds like it would be complicated? It's just DNA that has fallen off or otherwise got left behind by just about every living thing that lives or has visited an area of land, or that lives in a stream or other body of water like a pond, lake, or wetland habitat. Soil and water samples are taken and all of that DNA is then analyzed in a lab in order to know what creatures are or were present "out there". The idea here is to use the technology by taking soil or water samples ourselves to hunt for Bigfoot DNA. I'm trying to find a lab or two that will process the samples and we've been discussing the materials we need (that won't cost an arm and a leg) to extract field samples from the environmental habitats for testing in the labs.

 

@Madison5716 and @NorthWind Hey, you two are just great. Thanks :) 

Edited by hiflier
  • Thanks 1
  • Upvote 1
Link to post
Share on other sites
NorthWind
1 hour ago, hiflier said:

 

Isn't there kind of a famous name for your area? A "something" Triangle? Seems to me a saw a book somewhere about the strangeness that goes on around there.

 

And while "environmental" DNA sounds like it would be complicated? It's just DNA that has fallen off or otherwise got left behind by just about every living thing that lives or has visited an area of land, or that lives in a stream or other body of water like a pond, lake, or wetland habitat. Soil and water samples are taken and all of that DNA is then analyzed in a lab in order to know what creatures are or were present "out there". The idea here is to use the technology by taking soil or water samples ourselves to hunt for Bigfoot DNA. I'm trying to find a lab or two that will process the samples and we've been discussing the materials we need (that won't cost an arm and a leg) to extract field samples from the environmental habitats for testing in the labs.

 

@Madison5716 and @NorthWind Hey, you two are just great. Thanks :) 

Hopefully a lab with the ability to test for your "Notch" gene. And I, like Madison, am confident we will find prints. It will be interesting to see how much our "younguns" have grown.

  • Upvote 1
Link to post
Share on other sites
Catmandoo
9 hours ago, wiiawiwb said:

Here is an e-DNA aquatic kit for collecting a sample from JonahVentures. It uses a large syringe to push the water through the filter.

 

https://jonahwater.myshopify.com/

 

Jonah Ventures has  'diet test kits ' that  are used for herbivores and carnivores. Worth a look.  Send in a scat sample!   Turdology is looking up. They take PayPal.

  • Upvote 2
Link to post
Share on other sites
hiflier
2 hours ago, NorthWind said:

Hopefully a lab with the ability to test for your "Notch" gene

 

That's my hope as well, NW, but it is a different kettle of fish so to speak than the environmental mitochondrial DNA that most will collect in the field. Mitochondrial DNA is found outside of a cell's nucleus, so there's lots more of it to sample for and test. But the actual genes themselves are inside a cell's nucleus and located in nucleus's chromosomes. DNA-wise, there's not nearly as much of it as there is mitochondrial DNA. So to get a sample that has a nucleus with genes in it, one needs to extract a whole cell from the environment. A LOT of whole cells would be better of course ;) but genes from one good cell could be amplified (copied) by what is called PCR. then there would be enough gene material to test for the NOTCH2NL genes.

Link to post
Share on other sites
NorthWind
1 hour ago, hiflier said:

 

That's my hope as well, NW, but it is a different kettle of fish so to speak than the environmental mitochondrial DNA that most will collect in the field. Mitochondrial DNA is found outside of a cell's nucleus, so there's lots more of it to sample for and test. But the actual genes themselves are inside a cell's nucleus and located in nucleus's chromosomes. DNA-wise, there's not nearly as much of it as there is mitochondrial DNA. So to get a sample that has a nucleus with genes in it, one needs to extract a whole cell from the environment. A LOT of whole cells would be better of course ;) but genes from one good cell could be amplified (copied) by what is called PCR. then there would be enough gene material to test for the NOTCH2NL genes.

 

Holy smokes. Thanks for the education.

Link to post
Share on other sites
Twist
1 hour ago, hiflier said:

That's my hope as well, NW, but it is a different kettle of fish so to speak than the environmental mitochondrial DNA that most will collect in the field. Mitochondrial DNA is found outside of a cell's nucleus, so there's lots more of it to sample for and test. But the actual genes themselves are inside a cell's nucleus and located in nucleus's chromosomes.


Im confused Hiflier,  is the Notch gene found in E-DNA or not?  If I’m understanding correctly you state above that EDNA is going to produce Mitochondrial DNA.   You (below) state your gene is located in NuDNA.   

 

 

MyNOTCH2NL genes apparently are nuDNA genes(chromosomal) which I need to do researchmore on


 

I’m probably missing something but on the surface it seems confusing.

 

 

Link to post
Share on other sites
hiflier
Posted (edited)

Hopefully I can clear things up. An environmental DNA sample can contain either fragments of DNA or whole cells depending on many factors such as moisture, microbial activity like bacteria, the sun/UV light, soil pH and other things that can degrade DNA in Nature. This is true for soil. For water there is turbulence, again bacteria the water's pH and other things that will degrade DNA. The article I linked from the UK/Amazon studies sampled areas not unlike places in the US and they were very successful in detecting land animals in water samples.

 

What is true, though, is that actual whole cells are what would be needed to run testing for NOTCH2NL genes, or ANY genes for that matter. One needs a cell's nucleus with it's chromosomes in order to do that. An active BF area would mean a fresh supply of cells in which gene testing could be done.

 

The short answer is DNA samples that contain cells have a good chance for running nuclear DNA protocols that target specific genes. Period. Beyond that one is left with the more abundant mitochondrial DNA which are not genes but still have markers in small base-pair sequences in which species identifications can be made. Think Olympic Peninsula nesting site. Fresh fecal material would have yielded a LOT of cells for gene testing. But fecal material needs to be fresh as we all know. What got sampled was the soil beneath the nests and evidently there was a lot of degraded DNA but it was good enough to detect things like beas, elk, birds and other normal creatures including Humans. Evidently though, there were no whole cells available in order to test for genes. But there were still enough mitochondrial DNA there to determine different species. Degraded Human DNA was also present as we all now know.

 

So, whole cells will show genes, including the gene-bearing chromosomes inside a cell nucleus, and things degrade from there. Fortunately there is so much mtDNA that even small fragmented mtDNA base pairs can be run against the GenBank. I could get a lot more technical on markers but there's no need. Hope this helps.

Edited by hiflier
  • Thanks 1
Link to post
Share on other sites
BlackRockBigfoot
18 hours ago, Catmandoo said:

 

Send in a scat sample!   Turdology is looking up. They take PayPal.

Best comment of the day right here and nobody said anything...

 

"Turdology is looking up.  They take PayPal".   Lol.  

  • Upvote 1
Link to post
Share on other sites
MIB
MODERATOR
12 hours ago, hiflier said:

Hopefully I can clear things up.

 

Pretty nicely done.   

 

In your study so far, how concentrated do the cells need to be?   In other words ... ok, lets go with BobbyO's / Squatchermetrics' notion of bigfoot in headwaters.     The farther downstream you get, the more tributaries merge, so the better the chance the area upstream from the test site has bigfoots in it.   (Oversimplified, but .. true.)   On the other hand, the farther downstream, the more tributaries, the bigger the stream you'll be testing.    In what you've read so far, do there seem to be procedural suggestions about how far downstream it is reasonable to test and still expect to pick up cells thus DNA?  

 

Lets say .. to continue with BobbyO, who I think mentioned the Klamath River, if the bigfoots were on the east slope of the cascades fairly high up, would you start downriver by Klamath Falls where the river runs a couple thousand CFS?    The probability of cells somewhere in that is high, but is it too diluted?    Do you want to hit a tributary that has 1 cfs or less?  10 or less?  100 or less?    Or how big a drainage area upstream ... in square miles?   Or do you need to cut the area drained down to a few acres?

 

... yes, if you haven't guessed, a particular location (or two) come to mind.   

 

MIB

Link to post
Share on other sites
hiflier

Nope, never would have guessed that :) and thank you for the compliment. If I knew the specifics of where creatures are living then the answers to your questions would be easier to address. And since I don't know the conditions regarding cfm of any given stream it certainly outside my ability to even speculate. What else I don't know? Is in that April 2020 article about detecting land mammals in the UK and Amazon Basin it wasn't stated the conditions of neither the speed, size or depth of the streams and rivers that were sampled. Nor was anything said about how close or far away from a headwater the samples were taken. I could speculate that since quite a few different species were detected then perhaps the samples were extracted further downstream in a watershed?

 

I can give you an example of where I plan on taking samples, even though historically only a few sightings have ever been reported. The area is large and remote and so it could just be fewer people per square miles. It is in kind of a wet area surrounding a pond that collects runoff, and perhaps could even be somewhat spring fed. It is at elevation in a saddle between two relatively small mountains. From that wet area a stream flows down in into several waterfalls- each getting larger and faster as the terrain drops away. The last waterfall has a pool at the bottom and from there water exits the pool and flows into a much larger pond, into another pond and eventually into a large lake.

 

I'm sure that kind of a set up is everywhere in your neck of the woods. I think the risk of me contaminating any samples will be highest at that first pond between the two hilly drainages. So I will sample below that in one of the flows between the waterfalls. The further downstream the more area that will be draining into this water system. I also think there a trade off where the steepness of the terrain will feed any mammal DNA much quicker into a stream that is a much wider area where DNA will end up trapped in soil and never make it to open water.

 

In the sequel to my novel I am bring this dynamic out where one of the main characters has been waiting for weeks during very dry weather for a heavy rain to wash all the fresh and fragmented DNA into a pond. The goal being to detect all of the aquatic AND land mammals all at once (metabarcoding). Because, as we all know, until the ground saturates, such rains will initially flow on the surface as a heavy runoff. And BTW, just between you and me? my character doesn't know about the Bigfoots........YET! Or should I say YETI !! ;) 

 

 

Link to post
Share on other sites
NorthWind
3 hours ago, MIB said:

 

Pretty nicely done.   

 

In your study so far, how concentrated do the cells need to be?   In other words ... ok, lets go with BobbyO's / Squatchermetrics' notion of bigfoot in headwaters.     The farther downstream you get, the more tributaries merge, so the better the chance the area upstream from the test site has bigfoots in it.   (Oversimplified, but .. true.)   On the other hand, the farther downstream, the more tributaries, the bigger the stream you'll be testing.    In what you've read so far, do there seem to be procedural suggestions about how far downstream it is reasonable to test and still expect to pick up cells thus DNA?  

 

Lets say .. to continue with BobbyO, who I think mentioned the Klamath River, if the bigfoots were on the east slope of the cascades fairly high up, would you start downriver by Klamath Falls where the river runs a couple thousand CFS?    The probability of cells somewhere in that is high, but is it too diluted?    Do you want to hit a tributary that has 1 cfs or less?  10 or less?  100 or less?    Or how big a drainage area upstream ... in square miles?   Or do you need to cut the area drained down to a few acres?

 

... yes, if you haven't guessed, a particular location (or two) come to mind.   

 

MIB

 

I certainly don't know the answer to this, but I would imagine that if one had a filter and enough time to really get a lot of  filtrates for sampling, something interesting would show up. But you may run the risk of human contamination. You might have to go up higher into the less 
"populated" areas that humans visit less frequently. There's a lot of water in that river. Bound to be something. Good success to you, because I don't like the word "luck".

  • Upvote 1
Link to post
Share on other sites
SWWASAS

Has anyone seen a mention of the NA man that was tested for DNA by a relative?     A relative tested him using Ancestry.com.      Anyway I cannot remember his tribe but he was full blooded and his ancestry showed markers that took him back to the original peoples that supposedly made the Bearing crossing.     I was wondering what would happen if one of us got a good DNA sample of BF,   and simply submitted it to one of these ancestral DNA processors.     I had mine done by a National Geographic lab .   The testing lab would process it without discrimination.   The results certainly might be tossed out but might show something as remarkable is if Zana were tested.  It would be very interesting to see what the results would be.  

Edited by SWWASAS
  • Upvote 2
Link to post
Share on other sites

Create an account or sign in to comment

You need to be a member in order to leave a comment

Create an account

Sign up for a new account in our community. It's easy!

Register a new account

Sign in

Already have an account? Sign in here.

Sign In Now
×
×
  • Create New...