# What would you like to see researched?



## Fantastica (May 5, 2013)

I'm an undergrad at Umass Dartmouth and would really like to start doing dart frog / amphibian research. An immunology/ microbiology professor expressed interest in doung research related to Bd, but I don't think I need to limit myself to that specific scope. 

The thing is, the amount of possible research I could approach is pretty overwhelming. I'm wondering: what frog question would you like to see answered?


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## hypostatic (Apr 25, 2011)

Would this study be done in the field or in the lab in MA?


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## easternversant (Sep 4, 2012)

Does someone there have a colony of frogs? Because setting up a lab full of poison frogs for research would be A) hella expensive and B) hella time consuming.

Just sayin'.


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## frogparty (Dec 27, 2007)

I would like to see a more in depth study of the individual peptides and alkaloids present in the skin secretions and how they pertain to drug discovery.


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## Julio (Oct 8, 2007)

frogparty said:


> I would like to see a more in depth study of the individual peptides and alkaloids present in the skin secretions and how they pertain to drug discovery.


There are a few drugs made from Darts frogs, prob is that Pharmacudical companies have such tight security pretty much nothing is disclosed


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## frogparty (Dec 27, 2007)

I know about some of the research, but there's so much to learn still


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## Fantastica (May 5, 2013)

This would be in the lab for now, but I eventually want to pursue research in Costa Rica in the future.
I have 9 variabilis tads currently developing, there's a possibility of using them. My professor did discourage the use of live animals, though, so I think it'd be more something along the lines of using fecals or swabs from frogs at home, or from a guy he knows that does research on chytrid.

I think peptides/alkaloids might fall under biochem, I'm not too sure. Would you suggest I focus on Terribilis if I went that way?

I'll just throw out there that it was this article that jogged my interest:
Devastating Frog Fungus Triggers Cell Suicides | LiveScience
I just don't know how to go about experimenting on the fungus cell walls, it seems like it might be above my skill level at this point.


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## frogparty (Dec 27, 2007)

the one really neat drug I know of from darts, epibatidine, comes from tricolor I believe. 

Terribilis would be an interesting study subject. A lot is known about the complex alkaloids, but I doubt much has been done in the way of discerning the functionality of the peptides that even CB animals should produce. 

Yes, this would fall under the category of bio-chem. 

I think theres a lot of really interesting research to be done with BD. We know a lot about it, but theres a lot more to learn always, and any novel research that yields good data caould be very useful to conservation efforts for amphibians worldwide. 

You as an undergrad likely wont have a ton of time, or resources to throw at this project though, unfortunately, so Id focus on experimental design that would yield good data with the resources that you have available to you


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## hypostatic (Apr 25, 2011)

So, the human microbiota has a huge influence on our immune response and how we fight infections and disease, and even affects surprising things like cardiovascular health.

Maybe you could look at your frogs' poop and see how their gut fauna affects their health?


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## Fantastica (May 5, 2013)

Yeah, I feel like the Bd would be a better grad project.

Hypostatic, do you know of any articles off the top of your head that reference this? I wonder if Bd carriers like bullfrogs just happen to have better gut bacteria.


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## hypostatic (Apr 25, 2011)

Well, here's an article referencing the thing I said about cardiovascular health and gut bacteria. 

http://www.nature.com/nm/journal/v19/n5/full/nm.3145.html

I did a quick literature search, and I couldn't find anything about bullfrogs, chytrid, and gut fauna, but here are some papers you might find interesting

JSTOR: An Error Occurred Setting Your User Cookie
Physiology of Batrachochytrium dendrobatidis, a chytrid pathogen of amphibians
Activity of antimicrobial skin peptides from ranid frogs against Batrachochytrium dendrobatidis, the chytrid fungus associated with global amphibian declines
Enzootic and epizootic dynamics of the chytrid fungal pathogen of amphibians
Life cycle stages of the amphibian chytrid Batrachochytrium dendrobatidis - [email protected]
Experimental Exposures of Boreal Toads (Bufo boreas) to a Pathogenic Chytrid Fungus (Batrachochytrium dendrobatidis) - Springer
Effects of Chytrid and Carbaryl Exposure on Survival, Growth and Skin Peptide Defenses in Foothill Yellow-legged Frogs - Environmental Science & Technology (ACS Publications)
HOST PARASITE INTERACTIONS BETWEEN FRESHWATER PHYTOPLANKTON AND CHYTRID FUNGI (CHYTRIDIOMYCOTA) - Ibelings - 2004 - Journal of Phycology - Wiley Online Library


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## OrangeTyrant (May 12, 2011)

Things with live animals (even if you're just collecting swabs and fecals at home) get challenging when research and universities are concerned because then IACUC comes into play, which while it is obviously very necessary, is LOTS of paperwork and red tape. I would try to design something with environmental or retrospective samples if possible just because I think you'll have better access and less obstacles to your work.


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## smoyer (Jul 9, 2007)

Bd is an emerging concern, and may be a great opportunity to start laying groundwork for graduate work. there will likely be many folks that make an entire career out of researching and then mitigating the damage this fungus has/will cause. 

As far as undergraduate work, as others have said, the most significant part of any research is accurate data. and having a scientifically sound project is often harder than it appears. If you have buy-in from a prof, you may want to take advantage of his interest/experience.


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## cml1287 (Mar 29, 2012)

as others have said, i think it would be difficult to get a prof to start a project w/ live animals from scratch. I got extremely lucky - there's a prof at my university who is a behavioral ecologist and focuses on frogs. We have live frogs on campus that we use for research, but to study the poison aspects of darts in captivity would be difficult because they don't produce the toxins. 

I also think it's a little uncommon for undergrads to get actual hands-on research experience (unless you're really good - and you go to Dartmouth, so you just may be). Most undergrads at my university who are "involved" in research labs are basically just lab-hands, and not doing much in the way of actual research.


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## cml1287 (Mar 29, 2012)

easternversant said:


> Does someone there have a colony of frogs? Because setting up a lab full of poison frogs for research would be A) hella expensive and B) hella time consuming.
> 
> Just sayin'.


I don't think it would be too expensive, animals for research purposes aren't typically housed in the kind of elaborate enclosures you see here. Bare necessity are the primary concern, not how appealing the enclosure is.


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## easternversant (Sep 4, 2012)

Here is an idea involving both Bd and the hobby. Why don't you get people to swab their frogs and send you samples with their zip code. Then you could get an idea just how common Bd is in the trade, this could help with future legislation. Plus, your only overhead would be lab reagents. 

You may need to deal with both IACUC and IRB though, and you'll definitely want to make people totally anonymous.


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## frogparty (Dec 27, 2007)

Yeah if you order the right primers and have access to a PCR machine that's an easy project to do.


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## Mantella71 (Oct 7, 2013)

I conducted a study/analysis of egg production during wet/dry seasons between 2 species of dart frogs (azureus & auratus). Not surprisingly they both produced more during the wet season. Unfortunately I lost it back in 96 the year after I wrote it. My professor at the time, Dr. Forester (renowned salamander expert), said I should have it published. I think more studies should be done on effects of habitat loss and how dart frogs adapt or decline. Just a thought.


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## Dendrobait (May 29, 2005)

cml1287 said:


> I also think it's a little uncommon for undergrads to get actual hands-on research experience (unless you're really good - and you go to Dartmouth, so you just may be). Most undergrads at my university who are "involved" in research labs are basically just lab-hands, and not doing much in the way of actual research.


I know of many undergrads, myself included, that have done full blown research projects. They have to be carefully picked to be inexpensive and easy enough to do with limited resources and time, yet still produce rigorous results. They do frequently piggyback on a grad students project(such as using specimens collected by a grad student). A lot of these projects remain unfinished...I would contact graduating undergrads who you know are involved in research labs and they might hand you a half finished project.


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## Pubfiction (Feb 3, 2013)

What can we feed frogs to remove the need for supplements.


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## Fantastica (May 5, 2013)

Thanks for all the awesome feedback, guys! I appreciate it!




hypostatic said:


> I did a quick literature search, and I couldn't find anything about bullfrogs, chytrid, and gut fauna, but here are some papers you might find interesting


Thanks Hypostatic, I'm excited to start delving into some of these now that I'm on break!



cml1287 said:


> I also think it's a little uncommon for undergrads to get actual hands-on research experience (unless you're really good - and you go to Dartmouth, so you just may be). Most undergrads at my university who are "involved" in research labs are basically just lab-hands, and not doing much in the way of actual research.


Unfortunately, I don't go to the Dartmouth you're thinking of  However, I was encouraged to do a basically fully independent research project, when I came in with the idea of learning how to analyze my own frogs for chytrid. So, I don't *think* it would be too far of a stretch for me to do testing with swabs, but I;ll for sure have to check.



easternversant said:


> Here is an idea involving both Bd and the hobby. Why don't you get people to swab their frogs and send you samples with their zip code. Then you could get an idea just how common Bd is in the trade, this could help with future legislation. Plus, your only overhead would be lab reagents.
> 
> You may need to deal with both IACUC and IRB though, and you'll definitely want to make people totally anonymous.





frogparty said:


> Yeah if you order the right primers and have access to a PCR machine that's an easy project to do.


I love this idea, and I think it's totally do-able. The only big problem is that we don't have access to a PCR machine  *Does anyone know of a way to determine the presence of Bd from swabs without PCR?*
Another question is, are you guys willing to send me samples (you can just like this post or send a message)? They would of course be anonymous in the report, separated by species or genus, to show the prevalence of the disease in specific groups.


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## Fantastica (May 5, 2013)

So I decided to go the route of testing everyone's frogs for Chytrid. I just got out of my proposal meeting and he said he basically had everything.

There's two things I need to research and would appreciate any help:

1. I need a positive and a negative chytrid test. The negative test I'm assuming I can get from someone who's treated their frogs and recently got a negative chytrid report. The positive is the issue. I need a DNA or cDNA sample of chytrid to test. It would probably need to be local-ish (I live in MA).

2. I need to identify the sequence of nucleotides that people are using for testing for chytrid, so I can get the right primers. The professor recommended I look at scientific articles, and that's what I'll be spending most of my day doing.


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## hypostatic (Apr 25, 2011)

i actually posted the oligos for genotyping for chytrid recently. let me see if i can find them again...

here ya go:
http://www.jwildlifedis.org/content/40/3/420.full

Bd1a:
5′-CAGTGTGCCATATGTCACG-3′

Bd2a:
5′-CATGGTTCATATCTGTCCAG-3′ 

Also found a qPCR for detecting chytrid levels
http://www.savethefrogs.com/threats/chytrid/images/pcr-protocol.pdf


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## Fantastica (May 5, 2013)

Thanks so much! I just found that one. I also found another that uses ITS1-3 Chytr and 5.8S Chytr, I'm wondering which one is more widely used.

http://www.int-res.com/articles/dao2004/60/d060p141.pdf

I actually referenced that protocol in my proposal and he told me I absolutely don't want to use qPCR, since that will tell me how much Bd instead of just straight up positive Bd / negative Bd. Basically more complicated and more expensive.



hypostatic said:


> i actually posted the oligos for genotyping for chytrid recently. let me see if i can find them again...
> 
> here ya go:
> http://www.jwildlifedis.org/content/40/3/420.full
> ...


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## hypostatic (Apr 25, 2011)

Yeah, for example, you could use qPCR to find how how bad an infection is in an individual. And yes, qPCR is a much more comlpex and expensive endeavor.

I think a good way (other than actually surveying all the researchers working with Bd) to figure out which is more commonly used is to do a literature search on google scholar and see how many citations each gets.

For "Bd1a Bd2a" I got a combined 219 citations for the top two papers.

For "ITS1-3 Chytr 5.8S Chytr" I got a combined 104 citations for the top two papers.

This paper seems to use both:
Amphibian chytridiomycosis in Japan: distribution, haplotypes and possible route of entry into Japan - GOKA - 2009 - Molecular Ecology - Wiley Online Library

Oh and here's another link to the first paper I mentioned with bd1a/bd2a (which is also the most cited paper). I get a 404 for some reason on my end from the link I provided, but this link seems to be working:
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or if THAT doesn't work, here it is on pubmed
http://www.ncbi.nlm.nih.gov/pubmed/15465708


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## InvertaHerp (Mar 4, 2013)

Has something ever been done to determine how they synthesize their prey (wild/poisonous) to create the alkaloids? Perhaps look at their poo to look into gut bacteria etc


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