Specifically, I curious is anyone has experimented with different dye concentrations, staining duration's, and how long the larvae remained distinguishable from non-stained individuals. The literature has all sorts of nasty stories about dye concentrations that work on one species and causes 100% mortality in another, something I'd like to avoid if possible.
Cheers,
Paul
Well, many years ago I tested the concentration with methylene blue in Physalaemus cuvieri tadpoles. I diluted in water at a concentration of 10% and left for tadpoles for 5 min. Painted tadpoles kept the "marking" for 30 minutes in aquarium with water. Thereafter, tried to increase the concentration to 25% and after 5-7 min tadpoles showed lethargy and one of the individuals died.
I believe that if you use another compound, possibly painting can last a longer time.
Hi, I don't have experience with dyes, but I suggest you the use of visible implant elastomers (VIE). They are easy to use in larvae and last for years. Have a look here http://www.pwrc.usgs.gov/marking/vie.html and here http://ufwildlife.ifas.ufl.edu/pdfs/campbell%20et%20al%20%202009%20aphe.pdf about the use of VIE in frogs. I also recommend you a recent paper by Mike benard showing the use of elastomers on wood frogs http://onlinelibrary.wiley.com/doi/10.1111/gcb.12720/full
Good luck
Thanks Ibere and German, especially for the papers.
Unfortunately, my project is not about tracking the individuals over several years, I'm interested in comparing how different larval sample techniques perform to give a estimate of the total number. Therefore I only need to mark them temporarily (4-8 hours) and use a field technique that is easy and cheap.
Cheers,
Paul
You might consider staining with calcein; that works pretty well with zebrafish at least and toxicity is supposed to be pretty low. After staining my fish they are distinguishable for at least a week. You'll need a blacklight to see the stain well though.
http://www.nightsea.com/articles/detect-calcein-stained-fish/
http://onlinelibrary.wiley.com/doi/10.1111/j.1439-0426.2010.01418.x/abstract
We used a dye bath colouration with Bismarck Brown©. Concentration and handling referred to Rotary Screw Trap Protocols 15th of March 2006. Four days are possible (tested in gobies)
Easy and cheap
Greetrings
Jost
I wanted to post this follow-up as I've now done some mark-recapture work with wood frogs and spring peepers using neutral red stain. The concentration and exposure time from Herreid and Kinney (1966)
http://www.jstor.org/stable/1935651?seq=1#page_scan_tab_contents
of 0.02% for 30 mins killed or incapacitated most of the batch of wood frog larvae. The 0.002% concentration for 10mins from Yung et al (2002)
http://www.bioone.org/doi/abs/10.1670/0022-1511(2002)036%5B0465:AEOPIA%5D2.0.CO%3B2
was good but we extended the time to 20 mins to get a better stain and it worked great.
Cheers,
Paul
I don't know if it is still usefull, but I use VIE for larvae, and they retain it after metamorphosis. Applied for several tropical species and worked just fine for over 5 years already.
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