# How much of a problem is this exposure to EDTA to my frogs?



## Lordoftheswarms (Apr 27, 2009)

I have two main solutions that I water my tanks with. It was suggested that I ask what effect these may have on the frogs. I've had them for a week, and have seen no adverse effects on the frogs. I have noticed the toes of the Azureus twitching a little bit, but I haven't attributed that to the fertilizer yet, and don't know if that is a bad sign or not. My Typhlonectes natans caecilian didn't like it very much when I used the same bucket to do a water change that had previously had waste water that I drained from my vivaria watered with these solutions. I did rinse it out, but I noticed a slight change in behavior by the caecilian. At the time, this was attributed to the different water quality. I used a different bucket, and the behavior went back to normal.

The solutions are as follows. Presumably, the elements are not just elements, but compounds of sulphates or hydroxides, oxides, etc. My guess is that the solution is a buffer of the component acids and bases. My only concern is with the long term exposure to EDTA.

1.) 5 ml/8000 ml 0-0-3
SIO2 6.66%
K 2.55%

pH Down (to reduce the pH of the above solution to roughly neutral)
1 ml/8000 ml 0-26-0
K3PO4 18.28%

2.) 3 ml/8000 ml 5-0-0
Ca 3.43 %
N (T) 4.8 % (not sure what this is)
NO3 3.62 %
NH4 1.18 %

3 ml/8000 ml 0-9-6
P 3.92 %
K 4.96 %
Mg 1.62 %
S 2.13 %

3 ml/8000 ml 1-0-1
N (T) 1.38 %
NO3 0.59 %
NH4 0.59 %
K 0.39 %
Mg 0.17 %
Fe 0.11 %
Mn 0.03 %
Zn 0.006 %
Cu 0.0051 %
B 0.02 %
Mo 0.0009 %
EDTA 0.63 % (See below)
DTPA 0.20 %

MSDS for EDTA:
http://www.ee.iitb.ac.in/~nanoe/msds/EDTA.pdf

According to wikipedia:
EDTA is in such widespread use that it has emerged as a persistent organic pollutant.[23] It degrades to ethylenediaminetriacetic acid, which then cyclizes to the diketopiperizide, a cumulative, persistent, organic environmental pollutant. An alternative chelating agent with fewer environmental pollution implications is EDDS.
EDTA exhibits low acute toxicity with LD50 (rat) of 2.0 – 2.2 g/kg.[4] It has been found to be both cytotoxic and weakly genotoxic in laboratory animals. Oral exposures have been noted to cause reproductive and developmental effects.[9] The same study by Lanigan[9] also found that both dermal exposure to EDTA in most cosmetic formulations and inhalation exposure to EDTA in aerosolized cosmetic formulations would produce exposure levels below those seen to be toxic in oral dosing studies.

So what do you guys think? How much of a problem is the EDTA? 
(0.63/100) x 3 ml = 0.0189 mL / 8000 ml = 0.0000023625 ml EDTA/ ml of solution.
This solution is alternated with the first solution that doesn't contain EDTA, so it's about half that.

It was suggested that the plants, microbial life, and mycorrhiza would help eliminate the EDTA. Is that true? I'm going to look up if EDTA, and it's products can be metabolized.

Metabolism of EDTA:
Metabolism of Ethylenediaminetetraacetic Acid (EDTA) by Chickens

Apparently it can be digested, atleast by chickens, and later on in the paper, it mentions people:

Zn65 and EDTA-2-C14 were administered to 2 colostomized laying hens
in a metabolic chamber which permitted the collection of CO2 and excerta. EDTA-2-C14
was absorbed from the gastrointestinal tract and C14 from it was recovered in the
respiratory CO2, urinary uric acid and liver fat.

Improves the utilization of tranisition metals in the diet:

Kratzer et al. (3) reported
better utilization of zinc not only by using
autoclaved isolated soybean protein but
also by the addition of EDTA to the diets.
EDTA improved the utilization of zinc,
manganese and copper by chickens from
a diet containing isolated soybean protein
(4, 5). Scott and Ziegler (5) postulated
the existence of natural chelates in some
food materials which also improved the
utilization of zinc. Forbes (6) reported
improved utilization of zinc by rats when
the diet contained EDTA.

Wikipedia entry on DKP, no details on toxicity:
Diketopiperazines (DKP) are a class of cyclic organic compounds that result from peptide bonds between two amino acids to form a lactam. They are the smallest possible cyclic peptides.
A diketopiperazine was the first peptide to have its complete three-dimensional structure described, in work undertaken at Caltech by Robert Corey in the 1930s. Corey studied the cyclic anhydride of the dipeptide glycylglycine.[1]
Diketopiperazines are commonly biosynthesized from amino acids by different organisms, including mammals, and are considered to be secondary metabolites.[2] Some protease enzymes, such as dipeptidyl peptidases, cleave the terminal ends of proteins to generate dipeptides which naturally cyclize to form diketopiperazines.
Due to their rigid structure, chiral nature, and varied side chains, diketopiperazines are an attractive scaffold for drug design.[2] Both natural and synthetic diketopiperazines have a wide variety of biological activities including antitumor,[3] antiviral,[4] antifungal[5] and antibacterial[6] activities.

DKP can at least be excreted in the Urine, and "It is concluded that the treatments were without toxic effect." at the doses they were using, in rats:
ScienceDirect.com - Toxicology - Toxicity of aspartame and its diketopiperazine for Wistar rats by dietary administration for 104 weeks

Apparently Zn and Cu can be toxic as well, how EDTA impacts their toxicity is discussed in the paper, but I don't have an account to read the whole thing:
[]Man and Biosphere[] — Studies on the Sikkim Himalayas. Part 4: Effects of Chelating Agent EDTA on the Acute Toxicity of Copper and Zinc on Tadpoles of the Frog Rana hexadactyla - Khangarot - 2006 - Acta hydrochimica et hydrobiologica - Wiley 

"Approximately 44% of the total DKP excreted was excreted in the first 4 hr after dosing." On a tangent, the digestion of Aspartame gives DKP and METHANOL. Yikes. Methanol isn't something that you want in your body on a regular basis. Anyway: 
ScienceDirect.com - Food and Chemical Toxicology - Plasma and urine diketopiperazine concentrations in normal adults ingesting large quantities of aspartame


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## goof901 (Jan 9, 2012)

i have no idea what EDTA or what it is used for. but regarding your azureus' toe twitching, that is NOT caused by EDTA. they twitch their toe because it has to do with them hunting because the movement causes the insects to move, which result in the insect getting eaten.


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## WendySHall (Aug 31, 2010)

You know...I am absolutely not scientific whatsoever so can't really comment wisely on your question. However, words like "pollutant", "toxicity", and "reproductive and developmental effects" would really make me shy away. Why take the chances? Why not use a simple bottle of distilled water?

Btw...toe tapping is completely normal for darts.


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## goof901 (Jan 9, 2012)

WendySHall said:


> You know...I am absolutely not scientific whatsoever so can't really comment wisely on your question. However, words like "pollutant", "toxicity", and "reproductive and developmental effects" would really make me shy away. Why take the chances? Why not use a simple bottle of distilled water?
> 
> Btw...toe tapping is completely normal for darts.


agreed. plus there is no need for fertilizer because frog poo is great fertilizer


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## bsr8129 (Sep 23, 2010)

I have tried to read this and have no clue what you are even asking.


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## WendySHall (Aug 31, 2010)

My take on it is that he is misting (watering) his tank with two different solutions and that one contains some type of fertilizer containing EDTA (as well as some scientific letter combinations I don't understand) which may be harmful or lethal to frogs.

OP...Goof is correct, you don't need to add any fertilizers...the frogs take care of that themselves.


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## parkanz2 (Sep 25, 2008)

EDTA is an extremely common anti-coagulant and heavy metal chelator used in human and veterinary medicine. That being said, it's probably not a good idea to continuously expose your frogs to it or subject them to any high doses.

Edit: like others have said, it's probably just easier to use it RO or distilled water.


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## Lordoftheswarms (Apr 27, 2009)

Thanks for the responses guys. Before I kept frogs, plants didn't do so well on straight RO, so I added these solutions to keep the plants happy. 

I was looking for a number that would be a safe dosage for the frogs, or a definite answer one way or the other if it was safe to use. 
"Probably safer, and why take the chances" was what I was expecting, but it's not exactly what I was hoping for. Thanks anyways.


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## goof901 (Jan 9, 2012)

Lordoftheswarms said:


> Thanks for the responses guys. Before I kept frogs, plants didn't do so well on straight RO, so I added these solutions to keep the plants happy.
> 
> I was looking for a number that would be a safe dosage for the frogs, or a definite answer one way or the other if it was safe to use.
> "Probably safer, and why take the chances" was what I was expecting, but it's not exactly what I was hoping for. Thanks anyways.


aas previously stated, if you don't have frogs, then you would need fertilizer, but if you do have frogs, then you don't need fertilizer as their poo makes great fertilizer.


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## rachel1 (Apr 12, 2012)

In order to determine an exact number, you would have to find the molecular weight of EDTA and convert the ml's you are spraying to grams of EDTA. LD50 of 2g/kg means it wouldn't take very much to kill a tiny frog. If your frog is 1g, it would take .002g to have a 50% chance of killing the frog. 
I don't like the "cumulative, persistent environmental contaminant" part, as your viv is a closed system, and a cumulative contaminant will build up quickly. 
I also don't like the part about it being biologically active.
Is it necessary?


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## Lordoftheswarms (Apr 27, 2009)

The reason why I use it is because I've heard from more than a few plant fanatics that RO water will leach the micronutrients like Zn, Cu, Mn, Mg, Ca, Si, S, B, K, etc., and those aren't easily replaced quantifiably with an addition of tap water, or produced in any significant quantity from the frogs. All of the orchid resources said to dilute the fertilizer to a fraction of the suggested dose, so that's why I have it at 1/8th the suggested dose.

From another forum:



FrogPimp said:


> Nothing to worry about.
> 
> EDTA is the main ingredient in water dechlorinator solutions for fish that is responsible for neutralizing heavy metals. Heavy metals are never removed from solution, they are simply sequestered by the EDTA and are thus not available for further reaction. EDTA also binds Ca and Mg thereby reducing water hardness.
> 
> EDTA is one of my favourite molecules!


Based on the above, I'm not too worried, but I know that my Caecilian can't have dechlorinator, as they are ultrasensitive, so I will continue to investigate. It's not the molar weight I need, it's the density. From that, I calculate the grams. 

Do you guys have any objections to anything else in the list?


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## Lordoftheswarms (Apr 27, 2009)

Double post?


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## Lordoftheswarms (Apr 27, 2009)

0.0000023625 ml EDTA/ ml of solution

Density is 0.86 g/cm^3

0.0000023625 ml EDTA x 0.86 g/cm^3 = 0.00000203175 g /ml of solution

0.002 g = estimated 50% LD


0.00000203175/0.002 g = 984.37307739633321028669865879168 

I would need the EDTA from 984.37 ml of my solution to reach the estimated LD 50%. 
I do alternate with the other solution, so really, I would need the EDTA of about 2 L of my total irrigation solution to reach LD 50%

Seeing as this term has caused some concern, here is what it means: 
Persistent organic pollutants (POPs) are organic compounds that are resistant to environmental degradation through chemical, biological, and photolytic processes.

I had other people tell me that EDTA was metabolically decomposeable to CO2 and DKP, which is eliminated with the animal's uric excretions, so I'm not so sure I should be trusting the wikipedia article.


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## heyduke (Sep 19, 2006)

If the plants are that much of a concern then maybe do a terrarium without animals in it. And a Vivarium for the frogs with less sensitive plants.

Myself I like the frogs more than the plants. If the plants don't do well in the environment that my frogs do best in then I don't use them. I like leaf litter best of all. Good for the frogs and I can't kill it.

** This by no means implies that I hate plants in any way shape or form. I like them very much **


Sean


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## SOswanski (Mar 6, 2011)

Lordoftheswarms said:


> The reason why I use it is because I've heard from more than a few plant fanatics that RO water will leach the micronutrients like Zn, Cu, Mn, Mg, Ca, Si, S, B, K, etc., and those aren't easily replaced quantifiably with an addition of tap water, or produced in any significant quantity from the frogs. All of the orchid resources said to dilute the fertilizer to a fraction of the suggested dose, so that's why I have it at 1/8th the suggested dose.
> 
> From another forum:
> 
> ...


Sounds like you have likely looked at all different types of variables, so I don't want to sound insulting, but what type of lighting are you using and which plant types? If you edit those variables, you shouldn't have much of a problem finding a setup that is perfect for you. Also, if fertilizer is a must, try looking into organic hydroponic fertilizer. They have all the mineral elements you are looking for plus amino acids instead of chilates (sp?) and other additives which could be causing issues with your amphibs. 

Sometimes getting so focused on a single variable will prevent one from seeing the real issue, so perhaps set back and approach it one more time as a whole.


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## hypostatic (Apr 25, 2011)

Don't use chemicals in your frog tank. I use EDTA in the lab for frotein purifications, and I wouldn't want any chemicals near my forgs.

Amphibians BREATHE through their skin, so spraying them with any concentration of EDTA is like spraying their lungs with EDTA. For comparison, would you want to breathe in an inhaler with EDTA?

It might be alright to use it with plants, but your amphibians are extremely sensitive to any chemicals that end up in its environment. You said that your Caecilian can't have dechlorinator (which has been rated safe for aquarium fish I'm assuming?), so I suggest treating your frogs with the same chemical sensitivity for the best results.


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## Ed (Sep 19, 2004)

Lordoftheswarms said:


> The reason why I use it is because I've heard from more than a few plant fanatics that RO water will leach the micronutrients like Zn, Cu, Mn, Mg, Ca, Si, S, B, K, etc., and those aren't easily replaced quantifiably with an addition of tap water, or produced in any significant quantity from the frogs. All of the orchid resources said to dilute the fertilizer to a fraction of the suggested dose, so that's why I have it at 1/8th the suggested dose.


Why not use a clay containing substrate since a lot of those ions can be found in clay at levels needed for plant growth. Since clay binds those ions to some extent (either within the matrix and/or charge) that will reduce or prevent loss due to the RO or distilled additions. Loss of those ions seems to really only be a risk when using soilless substrates like ABG mix (and that is before we get into the additions that occur when adding dusted fruitflies (since all of those are included in the better supplements) which ends up in the system.... 


Ed


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## jeeperrs (Jan 14, 2010)

Ed said:


> Why not use a clay containing substrate since a lot of those ions can be found in clay at levels needed for plant growth. Since clay binds those ions to some extent (either within the matrix and/or charge) that will reduce or prevent loss due to the RO or distilled additions. Loss of those ions seems to really only be a risk when using soilless substrates like ABG mix (and that is before we get into the additions that occur when adding dusted fruitflies (since all of those are included in the better supplements) which ends up in the system....
> 
> 
> Ed


I use ABG mix. When I feed my frogs I use a little distilled water and rinse out the feeding cup to get the remaining vitamins (prevent waste). I then pour it into the substrate. I probably have the equivalent of miracle grow soil (that is a joke) in my tank with all the vitamins and calcium it gets from the waste vitamins, as Ed mentions. I mean, I throw away more vitamins/caclium than I can possibly use. So, might as well let the soil have some of it.


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## parkanz2 (Sep 25, 2008)

You want to be careful doing that though because some inverts can sequester certain types of nutrients far beyond what is required or healthy for a frog. I can't remember the details now, but I think vitamin E can be sequestered in isos at many hundred times the requirement for frogs. Those specifics could be totally wrong though... studying for finals and brain is shot. Hopefully Ed will be back to this thread to clarify.


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## skanderson (Aug 25, 2011)

another thought if you want to mist with a low dose fertilizer would be looking into using something like kelp tea, or fish emulsion. you can find both at any pot growing supply store. they seem to have a better assortment of fertilizers than the garden centers near me. one other thing ive noticed is that all the ferts in the pot stores are in much brighter containers that are almost psychodelic in appearance.


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## Ed (Sep 19, 2004)

parkanz2 said:


> You want to be careful doing that though because some inverts can sequester certain types of nutrients far beyond what is required or healthy for a frog. I can't remember the details now, but I think vitamin E can be sequestered in isos at many hundred times the requirement for frogs. Those specifics could be totally wrong though... studying for finals and brain is shot. Hopefully Ed will be back to this thread to clarify.


Fruit flies can sequester vitamin E (tocopherols) to levels far in excess of the levels in the food source to levels that can actually disrupt uptake of vitamin A and D3. Isopods can also sequester it to high levels (vitamin D3 is used as a cholesterol precursor so it is broken down, and vitamin A isn't utilized beyond what is needed to make the rhodopsin in the eyes). 

Ed


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## ShelbyFFS (Apr 30, 2012)

EDTA, as a chelation agent, binds to metals. Hence, it does bind to calcium. Since we supplement calcium to our frogs, I'm not sure what effect we'd see.


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## cbreon (Apr 25, 2005)

Ed said:


> Fruit flies can sequester vitamin E (tocopherols) to levels far in excess of the levels in the food source to levels that can actually disrupt uptake of vitamin A and D3. Isopods can also sequester it to high levels (vitamin D3 is used as a cholesterol precursor so it is broken down, and vitamin A isn't utilized beyond what is needed to make the rhodopsin in the eyes).
> 
> Ed


Many flake fish foods seem to have tocepherols, that combined with mite issues stemming from fish flake use make for a good reason to limit or eliminate fish-flake containing tocepherols as an isopod food...


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## Ed (Sep 19, 2004)

cbreon said:


> Many flake fish foods seem to have tocepherols, that combined with mite issues stemming from fish flake use make for a good reason to limit or eliminate fish-flake containing tocepherols as an isopod food...


As I noted above, it should really only be a concern if you are using the isopods as a feeder as opposed to stocking the tank with them. 

Ed


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## Scott (Feb 17, 2004)

FYI - I moved this bc/ we're basically questioning whether something will affect a frog's health.

s


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## cbreon (Apr 25, 2005)

Ed said:


> As I noted above, it should really only be a concern if you are using the isopods as a feeder as opposed to stocking the tank with them.
> 
> Ed


Sorry Ed missed that...also, was sort of commenting on my own transition away from fish food for this reason...


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## Ed (Sep 19, 2004)

cbreon said:


> Sorry Ed missed that...also, was sort of commenting on my own transition away from fish food for this reason...


Sorry, I was trying to keep the message clear since I've been quasi misquoted recently (not by you).. 

Ed


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## jeeperrs (Jan 14, 2010)

Ed said:


> Sorry, I was trying to keep the message clear since I've been quasi misquoted recently (not by you)..
> 
> Ed


So, can I quote you as saying the other member's quoting of you is a misquote? LOL!


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## ShelbyFFS (Apr 30, 2012)

Back to EDTA, I wonder if/ how much a frog would absorb and if it would be enough to disrupt calcium levels?


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## parkanz2 (Sep 25, 2008)

Ed said:


> Sorry, I was trying to keep the message clear since I've been quasi misquoted recently (not by you)..
> 
> Ed


My apologies if you're referring to me. I stated that I definitely wasn't clear on the details and that it needed further clarification (that you then provided!)


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