Biology Term paper advice: Chytrid

I have to start a term paper as the end of the year project for my 9th grade Bio class. The paper will be in the format of a "mock" Grant proposal. We are allowed to do the paper on any topic we like, as long as we have a good hypothesis (ex. "Captive penguins lay more eggs than wild ones" or "studying animal reactions to natural disasters"). Since I would like to do something Caudata related I was thinking I may do the paper on chytridomycosis A.K.A chytrid; however, I have some questions. First of all, I know Chytrid affects anurans, but does it affect newts and salamanders? I looked on the internet a bit but I keep coming up with things about Xenopus and other carriers. Also, I can't think of a good hypothesis involving the disease, can anyone help? If it doesn't effect newts and salamanders does anyone one have any other ideas? I would greatly appreciate any intelligent advice I can get!

P.S.
Since I have only ever read about the disease, does anyone know the proper pronunciation?

I've always heard and said "Ki-trid"

Chytrid affects all amphibians.

Xenopus and bullfrogs are well-known carriers.

You could look at how chytrid kills animals. Or you could look at how carriers spread chytrid (such as Xenopus being dumped into waterways after research). Or how researchers have actually spread chytrid by not sterilizing equipment properly.

You can't find any information about whether caudates are infected by chytrid or not... that sounds like the foundation of a hypothesis right there. Apply for a "grant" to investigate whether or not caudates are infected. I imagine you could pursue this by lab experiments or by testing wild populations.

Pronunciation:
chytrid (KIT-rid)
chytridiomycosis (kit-RID-ee-oh-my-KOE-sis)

Thank you both so much!!! I really appreciate the help. This is just the help I was looking for,this forum really is fantastic! Kaysie, Jen-you both rock.

Be sure to let us know what you end up writing!

I'd be interested to hear your hypothesis as well when you decide on it.

I seem to recall Ed Kowalski and others mentioning imported salamanders of the genus Bolitoglossa from Central and South America being infected with Chytrid.

As for a hypothesis how about something that's related to why certain species are carriers while others die so quickly? The effects of reinfection upon animals that have been treated with anti-fungal cures? Bridging off what Jen said you could use multiple testing methods to determine what is the best test for detecting chytridiomycosis. I know that polymerase chain reaction (PCR) and enzyme linked immunosorbant assays (ELISA) are used currently and I bet there are other methods like histological sections of skin.

I'd be more than happy to help you find or explain things. I love researching

Abrahm, in that case, I've got a 20 page term paper on how globalization has influenced the spread of wildlife disease...

If you don’t get an A+ with all this help I want to know why….

With regards to immunity, which I think is one of the most interesting areas of study; there are theories that some populations have symbiotic skin bacteria which defend against chytrid infection. I don’t know if anyone has determined why some populations have this skin micro fauna and others don’t but you could hypothesise about it - or even ways to spread this micro fauna to unprotected populations and save them all!

Here’s an interesting paper about this: http://www.jcu.edu.au/school/phtm/PHTM/frogs/papers/woodhams-2007.pdf

Kaysie said:

Abrahm, in that case, I've got a 20 page term paper on how globalization has influenced the spread of wildlife disease...

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Well, I can only promise results that are A+ for a 9th grade biology paper...

Plus I like working on a smaller scale.

I'm glad you changed the colors on your signature... talk about bright!

I'm still fiddling with the sig. ROY G BIV isn't doing it for me...

Wow, I'm so glad that all of you are so interested! Don't worry, when I'm done I will post a link to my paper on the forum, maybe even my rough draft so you guys can help with ideas. I turned in my hypothesis this afternoon, before reading the other posts, sorry Abraham! I decided to go along with Jen's suggestion of testing whether or not Caudates can be infected with Chytrid; However, if I can I may try and tweak it some later using more info. I will be sure to post anymore questions and keep you guys updated, I really appreciate the help! I'll have to kick myself if I don't get an A+ after talking with all of you.

P.S.
Abraham, I think if I knew what an "enzyme linked immunosorbant assays" was, my bio teacher would have to give me an A+ just for that.

Re: Rough Draft

Well, here it is, my very rough draft. I still am in the researching process, but this is what I have so far. I would appreciate any help on deciding on an experiment I could use to test my hypothesis. I have been thinking about doing something involving wild populations, but a lab experiment is possible. I also wasn't sure how well I "justified" my research. The assignment asks that I show the relevance of the research towards other scientists, the public or if not the public, scientific advancement in general. Any sources of additional info would be appreciated. Thanks in advance!

Just in case anybody is wondering, I found out that the project will be due on 5/19. My teacher revised my rough draft and said it was excellent, however I still need to come up with an experiment.

Nathan050793 said:

Abrahm, I think if I knew what an "enzyme linked immunosorbant assays" was, my bio teacher would have to give me an A+ just for that.

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Hey, you knew what PCR was, right? That's pretty awesome. Pregnancy tests are pretty similar to enzyme linked immunosorbant assays, actually, you just don't know the technical mumbo-jumbo.

Here is an abstract where a group of scientists actually found infected caudates. This would make a good base for constructing your experiment. Find caudates that inhabit areas that also have infected anurans and test the caudates.

Nathan050793 said:

Just in case anybody is wondering, I found out that the project will be due on 5/19. My teacher revised my rough draft and said it was excellent, however I still need to come up with an experiment.

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Hi Nathan,
I applaud you for wanting to undertake such an ambitious study. You are probably aware by now after further research, that at least 14 families and 93 species of amphibians have been diagnosed infected with Batrachochytrium dendrobatidis, inclulding caudates. Caudata species thus far recognized include: Amphiuma tridactylum, Necturus maculosus, Siren lacertian, Salamandra salamandra, Ambystoma tigrinum, Pseudotriton ruber and other plethodontid salamanders.

Setting up experiments would also seem to be an ambitious undertaking. Off the top of my head, if you were to do a lab-based setting, how would you obtain the pathogen? How would you culture the organism and what method would you use to infect the host? What pathogen 'load' (innoculation)would you use to try to introduce an infection? What specie of host or hosts would you use and at what study numbers...these factors could easily introduce a high degree of variability for results. At what point would you test for infection, how would samples be collected (swab, toe clippings , etc) and by what methods would diagnosis of infection be accomplished (polyclonal antibodies and immunoperoxidase stain, histology, etc)? All observation and diagnostic measures require an expertise. Diagnosis using histology requires skills in microscopy, a knowledge of the morphology of the chytrid, a knowledge of normal histology of the caudate skin, and an ability to differentiate the chytrid from other structures in the skin and other possible mycoses infections or contamination. Would the animals be euthanized or would you try to treat with antifungals (itraconazole, miconazone, etc) after inducing and diagnosing an infection?

Field experiments would also be challenging. There would have to be an identified Chytrid infected or carrying host population and then field experiments to test caudates inhabiting the same geography. Many collections over time would be needed. Mode of transmission (poorly understood) would introduce variability.

I know these are questions and not answers - but these are some of the issues that would need to be defined in the experimental methods.

I quickly scanned your proposal - I would be cautious of inferring the potential zoonotic potential of chytrid relative to human infections.

Best of luck with your research.

I created an experiment to test wild populations and a budget to go along with it. The experiment seems a little simple so if anyone ( Jan? Abrahm?) has any suggestions that would be appreciated. Also, I wasn't sure what the costs would be, so the budget is probably inaccurate (which is fine, it isn't required to be accurate at all, just an educated guess), so if anyone knows the actual costs or any expenses I left out that could be helpful. Enjoy!

Jennewt said:

Pronunciation:
chytrid (KIT-rid)
chytridiomycosis (kit-RID-ee-oh-my-KOE-sis)

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I've also heard it pronounced with a long i (like kite-rid); I think (though I'm not 100% sure) that the long i is more consistent with the Greek origin of the name.

Nathan050793 said:

I created an experiment to test wild populations and a budget to go along with it. The experiment seems a little simple so if anyone ( Jan? Abrahm?) has any suggestions that would be appreciated. Also, I wasn't sure what the costs would be, so the budget is probably inaccurate (which is fine, it isn't required to be accurate at all, just an educated guess), so if anyone knows the actual costs or any expenses I left out that could be helpful. Enjoy!

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Nathan - just a point of clarification - do you actually have to conduct this experiment?

jan said:

do you actually have to conduct this experiment?

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Oh, no! The point of the project was to create a possible experiment and to present it as if it were a grant proposal. I don't have to actually do the project- I think that my experiment would be nearly impossible for a 9th grade Bio-student to actually perform.

Caleb- that's how I thought it was pronounced too, however, most sources I have encountered pronounce it as Kit-rid.

Nathan050793 said:

I created an experiment to test wild populations and a budget to go along with it. The experiment seems a little simple so if anyone ( Jan? Abrahm?) has any suggestions that would be appreciated. Also, I wasn't sure what the costs would be, so the budget is probably inaccurate (which is fine, it isn't required to be accurate at all, just an educated guess), so if anyone knows the actual costs or any expenses I left out that could be helpful. Enjoy!

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Well, just to give you some ideas on your budget you list this:

$500.00 for Chytridiomycosis testing equipment

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If you wanted to buy the equipment you're looking at around $175 for 250 tests worth of qPCR mastermix, a mixture of nucleotides and Mg+ that are used to duplicate DNA. Then you're looking at about $2,000 dollars to $10,000 to buy a thermocycler to run the PCR in. You'd also have to buy qPCR primers and other supplies.

Some hobbyists send samples to outside labs for testing and are charged $25 per test. If you test just 50 animals that is $1,250.

Renting lab space is probably quite a bit more expensive too. Science is quite expensive (I'm typing this at work not more than 10 feet from a $750,000 mass spectrometer.)

I can totally get you a cheaper computer, though

As for your experiment, maybe you want to pick an anuran and name some species of caudates in the area. For example the mountain yellow-legged frog, Rana muscosa, and then use Amphibiaweb to map out its range and then use a combination of google and amphibia web to find the salamanders that share it.

Other things to include are how are you collecting the animals (i.e. sampling breeding adults from pools where infected anurans breed or turning over logs in the summer?) Are you just surveying an area and swabbing animals? Are you collecting them? How big a sample size (i.e. 2 salamanders or 50?) How many different species? Are you testing adults, juveniles, larva, or all stages? Are you even using qPCR or some other lethal technique (i.e. histological sectioning or SEM like the micrograph in your paper?)

With science it is all about details, details and more details.