# New Scientific Publication on PDF's and Carotenoids



## Allen Repashy (Jul 17, 2009)

Hey Everyone,

I just received an email from Justin Yeager at Tulane University to let me know that the first of several publications on their study of dietary Carotenoids and their effects on reproduction in PDF's has been published today.

I was lucky enough to be contacted by Justin when the project started about using our Calcium Plus for their studies and I have excitedly supplied them over the last few years and been anxious to read the papers.

Some really interesting information for sure. I was particularly surprised to see the results of supplementing the fruit fly media with high carotenoids. I wish I could have got them to use Superfly and SuperPig also, but when this project started, I hadn't finished developing Superfly yet, and they had been making their own media for years that they had great success with... They also used their own blend of carotenoid ingredients for the fly culture (SuperPig was also a relatively new product at the time). (the study started in 2009)

I have uploaded the paper to our website so you guys can view and download it. Interesting stuff indeed. Enjoy!

http://www.store.repashy.com/media/CERDF

Allen


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## Julio (Oct 8, 2007)

sweet! thanks for posting


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## hypostatic (Apr 25, 2011)

Huh, interesting. Of most interest, I think, is that frogs that did NOT receive carotenoid supplementation laid 1.75X more clutches!



> Although tadpole and metamorph output was higher, the number of clutches produced by females was lower under carotenoid supplementation. This decreased clutch production suggests that (i) oogenesis is not limited by carotenoid availability [see also Ogilvy et al., 2012], and (ii) reproductive (vs. tropic) egg production is suppressed when females are caring for tadpoles, as they were more often under the carotenoid supplemented treatment.


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## Allen Repashy (Jul 17, 2009)

Indeed, but it also states that the number of eggs per clutch was significantly higher, and the survival was higher, resulting in increased number of total offspring..... It makes sense the longer time between clutches allows for more transfer of nutrients to eggs......

Definitely not what you would expect.


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## Reef_Haven (Jan 19, 2011)

The amount of methylparaben in their media seems extreme to me. I thought the recommended amount was closer to 1 part per thousand. Anyone else using this much?
Maybe they misprinted the correct quantity, cause at least in this study that amount would be toxic to the fly larvae.
http://124.205.222.100/Jwk_spkx/EN/abstract/abstract11536.shtml


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## oldlady25715 (Nov 17, 2007)

Interesting analysis. 63 pairs of pumilio is quite a good sample size. 

I'd be interested in more info about the frogs. How long they were captive prior to the study--were they fresh off the boat or were they already kept for a period with the baseline media. Did they all have parasites? But I suppose as long as they were from the same import and quarantined uniformally, they constitute a suitable population for a study about ex situ breeding. 

I was a little surprised to see tap water on the list of ingredients, given the specific nature of everything else. 

Thanks for posting Allen,and I think your supplements and media work great.


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## frogfreak (Mar 4, 2009)

hypostatic said:


> Huh, interesting. Of most interest, I think, is that frogs that did NOT receive carotenoid supplementation laid 1.75X more clutches!


But,

"egg production is suppressed when females
are caring for tadpoles, as they were more often under the
carotenoid"


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## Allen Repashy (Jul 17, 2009)

I was asked off board about the Methylparaben levels used in the study compared to what I use in my Superfly product... I didn't realize how high they were until it was pointed out...... I thought I would go ahead and post my reply publicly because I think it might be of interest to most....

Hi XXXXX,

I didn't even notice that but yeah, that is a freaking LOT of methyl paraben.

I use a combination of six ingredients to control mold and bacteria: Calcium propionate, Malic Acid Cinnamon,Potassium citrate, methylparaben, Potassium Sorbate (In that order)

While I don't divulge exact proprietary information, I will say that the methylparaben levels I use would be measured in fractions of a percent.

On a dry matter basis, they appear to have used close to 10% methylparaben in their mix. When I originally started working on the Superfly, 90% of the real development work I did was learning how to control mold and bacteria...... nutritionally, fly media is a no brainer more or less. When you use a single preservative, you get great control over some types of growth and poor control over others..... So you just have to keep adding more until you control the difficult ones also...... often ending in levels that are very high like what you see here.....

My approach was to use different preservatives that each targeted different types of bacteria/mold...... and work in different conditions..... PH plays a huge roll in control also. Different preservatives/organic acids function at different PH levels..... For example, Sorbic Acid works very well at Low PH, but not so well at Neutral PH, where Propionic acid works better... I use Malic Acid and Citric Acid (from Potassium Citrate) to get the PH where I want it to be and keep it there using mineral buffers like the calcium and potassium....

Long story short, by first focusing on PH level and stability (which I think no other media manufacturers do) I first try to create the best environment for the preservatives to "work" in.... then by using different preservatives/organic acids that work on different types of molds and bacterias (for example, some ingredients work well with gram positive bacteria, some better with gram negative... some are better for black mold, and some better for the white fuzzy stuff)..... I can use much lower overall levels of preservatives, while dividing it again by a number of different ingredients to avoid any potential negative effects a single ingredient might contribute. 

So even using lower overall levels, the right combination of ingredients can still be more effective then super high levels of a single one. Methylparaben WILL kill it all..... but you can see the kind of levels necessary to do so..... more than three times total level of all the ingredients I use combined.... and it is the most toxic of all the preservatives IMHO.

Allen


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## Allen Repashy (Jul 17, 2009)

Just as a FYI, to create similar carotenoid levels to those used in the study (it is not apples to apples because I use a much broader spectrum of carotenoid ingredients compared to those in the study) I would estimate that in addition to the superpig levels already in the Superfly, that you would need to mix about nine parts Superfly with one part SuperPig to come up with a "similar" level of carotenoids in the media. This would creat a media higher in overall carotenoids than the study media, but have similar levels of astaxanthin..... I do need to check my math on this though because I am just doing this off the top of my head.

HOLD THAT THOUGHT.... I think I am off quite a bit here..... let me to some more math.. Be back soon.....................




OK, I ran some numbers through my software, and adding 5% (DMB) SuperPig to the Superfly should give us approximately:

Beta Carotene mg/kg 90.0000	
Astaxanthan mg/kg 50.0000	
Lutein mg/kg 135.0000	
Capsanthin 37.5000	
Zeaxanthin mg/kg 15.0000
Lycopene mg/kg 10.0000
TOTAL MEASURED CAROTENOIDS 337.5000	

A pretty comparable amount to the study..... over on some and under on others but overall, quite a bit higher in total carotenoids.





Allen


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## frogfreak (Mar 4, 2009)

This is very interesting as I feed a lot of larvae at times. 

Done yet?


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## SilverLynx (Aug 29, 2013)

Allen,

Thanks for posting the study. I am not surprised by the conclusions. I personally
use Superpig, Calcium Plus and Superfly I also add natural forms of carotenoids to the Superfly, as I am a firm believer that the gutload/media is extremely important and this study confirms that. I am interested in the amount of Superpig needed to replicate the carotenoid amounts used in the study.

Thanks,
Lane, aka, SilverLynx


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## Allen Repashy (Jul 17, 2009)

Hi Lane,

Scroll back a few posts about the SuperPig levels as I just went back and accurately calculated it out.

Allen


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## SilverLynx (Aug 29, 2013)

Allen,

I just saw the edit with the 5%. Keep up the good work!!! This scientist loves your products. Btw, I am seeing major tad development using your Community Plus. 

Thanks,
Lane, aka, SilverLynx


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## Reef_Haven (Jan 19, 2011)

So this study indicates there is a change within the flies that improved their nutritive value, that can't be addressed by simply increasing the amounts of carotenoids in the dusting supplement?


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## Reef_Haven (Jan 19, 2011)

Allen Repashy said:


> Hi Lane,
> 
> Scroll back a few posts about the SuperPig levels as I just went back and accurately calculated it out.
> 
> Allen


Superpig contains vitamin E, I thought we were concerned with flies accumulating Vitamin E to levels that would throw off the balance to Vitamins A and D?


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## Allen Repashy (Jul 17, 2009)

According to what I am reading, Calcium Plus was used for both groups, so if that was the case, then it is saying exactly that.

Could we duplicate this by dusting with more Superpig.... possibly so... If we mixed Calcium Plus with say 50% Superpig, that would give us a lot more carotenoids..... but it would cut the calcium and vitamins like Retinol, in half... SuperPig is also not what I would consider a micro-fine powder, so to get a good stick, you would need to take the time to grind it down finer.... 

There is also the issue that it is possible that the larvae are metabolizing the carotenoids into something that can't be provided by direct supplementation. 

Like every study I read, I walk away with some good answers, but I always feel like I get more new questions to ask than answers..... 

The study does contradict my presumption from other publications that it wasn't valuable to supplement drosophila with carotenoids, so that in it's self is a revelation and the main point to digest from this article IMHO.


Allen


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## Allen Repashy (Jul 17, 2009)

Reef_Haven said:


> Superpig contains vitamin E, I thought we were concerned with flies accumulating Vitamin E to levels that would throw off the balance to Vitamins A and D?


SuperPig does NOT contain any added vitamin E. Not sure where you got that from.

Ingredients below.... though I did recently decide to add another ingredient to SuperPig.. Watermelon Extract.... for it's high levels of the carotenoid Lycopene.

SuperPig
Carotenoid Supplement

Our Pigment Enhancement Formula is a Carotenoid Supplement designed to enhance the diet of Reptiles, Fish, Amphibians and Birds.

INFORMATION: Contains a “broad spectrum” of Carotenoids including, but not limited to: Astaxanthin, Capsanthin, Capsorubin, Beta-Carotene, Alpha Carotene, Beta Cryptoxanthin, Zeaxanthin, Neoaxanthin, Cucurbitaxanthin, Violaxanthin, Lutein, Echineone, Canthaxanthin and Lycopene.

INGREDIENTS: Calendula Flower, Pfaffia Yeast, Paprika, Marigold Flower, Algae Meal (Chlorella and Spirulina), RoseHips, Hibiscus Flower, Turmeric.


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## Reef_Haven (Jan 19, 2011)

Thank you Allen. Sent you a PM.


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## ecichlid (Dec 26, 2012)

Good stuff! On a practical level...

Is there enough Super Pig in Calcium Plus? Or should I dust with Super Pig on occasion? How often should I dust my melanogaster for my growing D. auratus? Can it be done too often?


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## Roadrunner (Mar 6, 2004)

So that means they can utilize carotenoids much better than previously thought by converting to vit. a? or do they contain another function in not being converted?

And I thought anything added to the medium in the form of carotenoids wouldn't be passed on to the frogs? Are they talking about medium fortification or dusting? 

Sorry, a little dry and I don't have the time to read thru just brushed the article.


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## Reef_Haven (Jan 19, 2011)

frogfarm said:


> Are they talking about medium fortification or dusting?


According to the study, the only difference between the control group and study group was the additions to the fly media. Both groups were dusted the same with Repashy Calcium plus and additional Vit A every 3 months.
What the study did not try to confirm was whether additional carotenoid dusting would have made similar improvements. 
The flies nutritive value definitely appears to have improved, but could the same effect have been shown with additional dusting is unknown.
Seems to support the argument for feeding a variety of different feeder insects.


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## hypostatic (Apr 25, 2011)

Reef_Haven said:


> Seems to support the argument for feeding a variety of different feeder insects.


Here's a section of the paper on that:



> The natural diet of O. pumilio consists primarily of ants and mites [Toft, 1995], with the latter the more carotenoid‐rich food source [Olson, 2006]. As a replacement for the natural diet, carotenoid supplemented fruit flies were superior to unsupplemented ones in our captive colony.
> Perhaps further highlighting the importance of prey item diet
> breadth, we found that wild insects attracted to fruit
> (primarily Drosophila spp.) allowed for successful reproduction by captive O. pumilio held outdoors at Bocas del Toro (unpublished data). Whether carotenoid supplementation addressed a vitamin A deficiency in this ex situ population and whether that deficiency limited reproductive success remain unclear.


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## Allen Repashy (Jul 17, 2009)

My interpretation is that when we consider the fact that the frogs should have been receiving more than ample Preformed Vitamin A (Retinol) from Calcium Plus AND a topical treatment, that the carotenoids are contributing other nutrients. 

Meaning no amount of Retinol supplement can replace the carotenoids because they are contributing different things. We should consider carotenoids as their own group of "vitamins" that can't be replaced by anything else. These compounds are really just starting to be studied and understood. 

After reading this, I have definitely changed my thoughts about the value of including significant amounts of carotenoids in the fly media. Superfly has SuperPig in it, but in low levels for two reasons. First, I was under the impression that it wasn't as important as this paper claims, and secondly, Superfly is already one of the most expensive fly medias on the market and adding a lot of superpig would just make most people think it was too expensive to even try.

If there was enough demand, I could make a Superfly + version, but I don't think it is that difficult to just add 5% superpig to the superfly for those who decide to follow this lead.. (it does seem to make a good argument for doing so) The other option of course is to dust more with carotenoids, but I am not so sure that would be the best approach because you would be "cutting" the other nutrients you are dusting with. I also think it is easier to load the media than dust with carotenoids because of the particle size issues and actual stick to drosophila of these ingredients.

Allen


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## JPccusa (Mar 10, 2009)

Allen Repashy said:


> If there was enough demand, I could make a Superfly + version, but I don't think it is that difficult to just add 5% superpig to the superfly for those who decide to follow this lead.. (it does seem to make a good argument for doing so)


I believe a "SuperFlyingPig" version would be ideal. I'm sure many people would switch from Calcium + to the new and improved version if the difference in price is not exorbitant.


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## Allen Repashy (Jul 17, 2009)

JPccusa said:


> I believe a "SuperFlyingPig" version would be ideal. I'm sure many people would switch from Calcium + to the new and improved version if the difference in price is not exorbitant.


Hmm.... I don't understand what you are saying, as modifying Superfly has has nothing to do with Calcium Plus formulation or usage.


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## SilverLynx (Aug 29, 2013)

I will be adding at least 5% SuperPig to to the Repashy Bug Burger and Community Plus. I put Bug Burger in all my Oophaga enclosures and grow out 
tanks, creating feeding stations. The flies congregate and feed on the Bug Burger. 

Thanks,
Lane, aka, SilverLynx


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## JPccusa (Mar 10, 2009)

Allen Repashy said:


> Hmm.... I don't understand what you are saying, as modifying Superfly has has nothing to do with Calcium Plus formulation or usage.


Sorry for the confusion. I meant to say Superfly instead of Calcium +.


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## Tincman (Jan 18, 2013)

This is pretty cool, we were just discussing Vitamin Supplementation in another thread & someone brought up the possibility of adding CoEnzymes/Vitamins to insects diet in order to create more nutritious feeders.. Its great that you posted this right now.. You are what you eat I guess... I wonder if there are any other ways to make flies more nutritious..?


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## Roadrunner (Mar 6, 2004)

So carotenoids can be passed on to the frogs thru enhanced media and not just thru what is found in the ff's eyes. Wasn't there talk about "gutloading" or further enhancing the flies nutritive value by additions to the medium? Didn't someone say it couldn't be done?


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## Allen Repashy (Jul 17, 2009)

frogfarm said:


> So carotenoids can be passed on to the frogs thru enhanced media and not just thru what is found in the ff's eyes. Wasn't there talk about "gutloading" or further enhancing the flies nutritive value by additions to the medium? Didn't someone say it couldn't be done?


I think Ed presented some publications that claimed that carotenoids were not stored by the flies...... but this seems to contradict this...... or some kind of "X" factor at least is involved, meaning the flys are converting the carotenoids into "something else" that has benefits......


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## ecichlid (Dec 26, 2012)

Allen Repashy said:


> My interpretation is that when we consider the fact that the frogs should have been receiving more than ample Preformed Vitamin A (Retinol) from Calcium Plus AND a topical treatment, that the carotenoids are contributing other nutrients.
> 
> Meaning no amount of Retinol supplement can replace the carotenoids because they are contributing different things. We should consider carotenoids as their own group of "vitamins" that can't be replaced by anything else. These compounds are really just starting to be studied and understood.
> 
> ...


 Thank you Allen! I will be purchasing SuperPig today. Your clarification opens up two more questions.

1) *If one owns SuperFly media*, how much SuperPig should be added to the media to make an ideal media (SuperFly+) in regards to carotenoids?

2) *If one owns fly media with out any supplements*, how much SuperPig should be added to the media to make an ideal media in regards to carotenoids?


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## JPccusa (Mar 10, 2009)

Answer 1 is here:



Allen Repashy said:


> ... I don't think it is that difficult to just add 5% superpig to the superfly for those who decide to follow this lead.. (it does seem to make a good argument for doing so)


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## hypostatic (Apr 25, 2011)

No offense to Allen, but I've been having the suspicion that the Repashy line has been deficient in vitamin A or carotenoids (or both?) for some time. (As far as the formulation relates to dart frogs)

This suspicion first arose when I noticed that people on the boards would often supplement Vitamin A Plus in addition to Calcium Plus, in order to get better breeding/eggs/tadpoles. Also, I've read that some people report that even though they've been dusting their frogs' food with Repashy Calcium Plus, some people's frogs still get short tongue syndrome, which is indicative of vitamin A deficiency.

The paper states that the primary diet of pumilio (and I think this also holds true for other dart frogs) is ants and mites -- and that these are a "carotenoid‐rich food source" (and *here* is the cited literature for that claim). Although while looking through the paper, the table that compares carotenoid content wasn't super clear on the claim that drosophila contains significantly less carotenoids than either ants or mites. Maybe someone else could take a look at the paper?

Anyway, my point is that I feel that the Repashy line would be much more complete for dart frogs if the vitamin A and/or carotenoid was increased.


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## Roadrunner (Mar 6, 2004)

I don't think the paper deals w/ vit. A or the conversion of carotenoids to vit. A. It just says that more carotenoids has an added benefit.

As far as I've seen I've never used a Vit. A supplement and have had a very hi success rate and very hi egg production rate have not had short tongue syndrome or spindly or other deformations at any significant rate.




hypostatic said:


> No offense to Allen, but I've been having the suspicion that the Repashy line has been deficient in vitamin A or carotenoids (or both?) for some time. (As far as the formulation relates to dart frogs)
> 
> This suspicion first arose when I noticed that people on the boards would often supplement Vitamin A Plus in addition to Calcium Plus, in order to get better breeding/eggs/tadpoles. Also, I've read that some people report that even though they've been dusting their frogs' food with Repashy Calcium Plus, some people's frogs still get short tongue syndrome, which is indicative of vitamin A deficiency.
> 
> ...


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## Allen Repashy (Jul 17, 2009)

hypostatic said:


> No offense to Allen, but I've been having the suspicion that the Repashy line has been deficient in vitamin A or carotenoids (or both?) for some time. (As far as the formulation relates to dart frogs)
> 
> This suspicion first arose when I noticed that people on the boards would often supplement Vitamin A Plus in addition to Calcium Plus, in order to get better breeding/eggs/tadpoles. Also, I've read that some people report that even though they've been dusting their frogs' food with Repashy Calcium Plus, some people's frogs still get short tongue syndrome, which is indicative of vitamin A deficiency.
> 
> ...


I think we are going to have to agree to disagree on this one.

I have moderate levels of vitamin A in the Calcium Plus, I already have a Calcium Plus HyD that has twice the levels of vitamin A as the regular version and one that has half the levels because many people "think" the regular calcium plus has too much. 

My Calcium Plus HyD is recommended for therapeutic use and use on basking species and true carnivores that are kept with little UVB .... and I do NOT think Dendrobates fall into this category or have this kind of requirement, so I will NOT recommend the Calcium Plus HYD for regular use with Dendrobatids. If you don't agree, then you, of course, are free to use my products however you want, or use other products.


I also make a Vitamin A Plus, which is a vitamin A only product for treating deficiency..... or periodic use with healthy animals. I think you should re read the posts about using Vitamin A Plus or another supplemental Retinol product because they are usually associated with animals that have had a history of deficiency... it that case, Calcium Plus can take quite some time to bring back deficient animals..... thus the additional supplementation. 

If you don't agree, then you, of course, are free to use my products however you want, or use other products that you think might work better. Do you really expect STS to completely disappear from every frog in every situation with all the variables out there... from varied usage levels, to using expired products..... to genetics..... to lack of real scientific research with multiple instances of the same cause and effect?



Allen


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## hypostatic (Apr 25, 2011)

Allen Repashy said:


> I think you should re read the posts about using Vitamin A Plus or another supplemental Retinol product because they are usually associated with animals that have had a history of deficiency... it that case, Calcium Plus can take quite some time to bring back deficient animals..... thus the additional supplementation.


Hmmm I guess I wasn't really taking animals with a history of deficiency into account, which would make sense why just calcium plus doesn't correct the problem.

But still, if you think the findings of the paper are important enough to consider adding more carotenoids to the fly media, why not the dusting supplement?


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## Allen Repashy (Jul 17, 2009)

hypostatic said:


> Hmmm I guess I wasn't really taking animals with a history of deficiency into account, which would make sense why just calcium plus doesn't correct the problem.


Yeah, it is an important factor and I have acknowledged that if an animal is deficient, that it can take many months for Calcium Plus to reverse the issue. But increasing the amount of A in calcium plus is not the answer, because then over the long term, we could have too much. This is the exact reason I decided to pull the trigger and put out the "Vitamin A Plus" as a product....



> But still, if you think the findings of the paper are important enough to consider adding more carotenoids to the fly media, why not the dusting supplement?


As I said in prior posts...... it is difficult to get a good stick when dusting with carotenoids because of the particle sizes, and if we are dusting with more carotenoids, then we have to be using less Calcium Plus..... there is always a give and take if you are already dusting at every feeding.

Adding it to the media solves this issue and any potential factors that could be involved dealing with the metabolism of the flies creating unique compuounds using the carotenoids that we aren't aware of..... but have value.

All theory really..


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## hypostatic (Apr 25, 2011)

I'm a bit frustrated that the authors didn't decide to like, grind up the two FF groups and analyze the contents to see what the actual difference inside the flies was. And --as I believe someone else mentioned-- that they didn't also try just adding more carotenoids to the dusting supplement to compare with adding it to the media.


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## frogfreak (Mar 4, 2009)

hypostatic said:


> I'm a bit frustrated that the authors didn't decide to like, grind up the two FF groups and analyze the contents to see what the actual difference inside the flies was. And --as I believe someone else mentioned-- that they didn't also try just adding more carotenoids to the dusting supplement to compare with adding it to the media.


That wasn't the purpose of the study and would take a new experiment to get the results. I'm not sure that would even be possible... 

I'm just glad they did this study. It's not everyday we get this kind of info! It's groundbreaking really, don't you think?

Glass half empty kinda guy?


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## Azurel (Aug 5, 2010)

Allen Repashy said:


> My interpretation is that when we consider the fact that the frogs should have been receiving more than ample Preformed Vitamin A (Retinol) from Calcium Plus AND a topical treatment, that the carotenoids are contributing other nutrients.
> 
> Meaning no amount of Retinol supplement can replace the carotenoids because they are contributing different things. We should consider carotenoids as their own group of "vitamins" that can't be replaced by anything else. These compounds are really just starting to be studied and understood.
> 
> ...


With the price of a 4oz bag of Superpig I don't think it would be to expensive to just get a second for the media....Based on the reading and posts here I will start adding it to my superfly.....Not to mention I hate throwing out amounts of suppliments seems a good way to make sure it all gets used up before the 6th month mark.

sent from my Galaxy S lll


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## edwardsatc (Feb 17, 2004)

hypostatic said:


> Although while looking through the paper, the table that compares carotenoid content wasn't super clear on the claim that drosophila contains significantly less carotenoids than either ants or mites. Maybe someone else could take a look at the paper?


The categories are quite broad in the table and they mix common names, orders, specific families etc ... not very well put together.

Anyway, according to their table both dipterans and ants have the same mean carotenoid content (120 mg/kg), but it does imply that mites (arachnids) have significantly more (280 mg/kg). 

That said, Dugas et al. only cite that paper in regards to the claim that mites are more carotenoid rich than ants. No claim is made as to a comparison between drosophila and either ants or mites (unless I missed it).

Their reference to Olson raises one of my pet peeves ... improper citation. Dugas et al.should have gone back to the original sources listed in appendix A of Olson. The table in Olson is neither a result or conclusion, it is merely introductory material composed of data extracted from other manuscripts of Olson. This is just pure laziness on the part of the authors ...


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## Ed (Sep 19, 2004)

There is another study using red-eye tree frogs that has some good information in it as well.. Sorry not free access see A brighter future for frogs? The influence of carotenoids on the health, development and reproductive success of the red-eye tree frog - Ogilvy - 2012 - Animal Conservation - Wiley Online Library for the abstract. 

Over the last couple of years I've increased the carotenoid (astaxanthin) content of the Calcium plus formula until it's about 50/50. I've seen better reproduction and an increase in red coloration of the pumilio. (And as a side note, that is with a once/week feeding of D. melanogaster and no other supplementation). I think I've seen all I can figure out using this method so will be switching up my trials in the near future. 
I saw a huge amount of froglets produced out of my caucheros. I think I'm up to nine froglets in the last 4 months from one pair (there is at least one extra in their cage now as I suddenly have two calling males again). 

Some comments 

Ed


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## Allen Repashy (Jul 17, 2009)

FYI, Mixing one four ounce bag of SuperPig with a four pound bag of SuperFly would give you 6% inclusion level

A 3oz jar would be just under 5% and since we are phasing out the 4oz bags at the end of the year, that would probably be the way to go.

Allen


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## Azurel (Aug 5, 2010)

Allen Repashy said:


> FYI, Mixing one four ounce bag of SuperPig with a four pound bag of SuperFly would give you 6% inclusion level
> 
> A 3oz jar would be just under 5% and since we are phasing out the 4oz bags at the end of the year, that would probably be the way to go.
> 
> Allen


Thanks Allen....and Ed as well....

sent from my Galaxy S lll


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## Ed (Sep 19, 2004)

edwardsatc said:


> That said, Dugas et al. only cite that paper in regards to the claim that mites are more carotenoid rich than ants. No claim is made as to a comparison between drosophila and either ants or mites (unless I missed it).


There is at least one study on carotenoid retention and storage by fruit flies see Giovannucci, David R.; Stephenson, Robert S.; 1999; Identification and distribution of dietary precursors of the Drosophila visual pigment chromophore: analysis of carotenoids in wild type and ninaD mutants by HPLC; Vision Research 39(2): 219-229

In general if I remember correctly from that study (and discussed here http://www.dendroboard.com/forum/food-feeding/67766-repashy-superfly-26.html#post608267) 

Some comments 

Ed


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## edwardsatc (Feb 17, 2004)

Ed said:


> There is at least one study on carotenoid retention and storage by fruit flies see Giovannucci, David R.; Stephenson, Robert S.; 1999; Identification and distribution of dietary precursors of the Drosophila visual pigment chromophore: analysis of carotenoids in wild type and ninaD mutants by HPLC; Vision Research 39(2): 219-229
> 
> In general if I remember correctly from that study (and discussed here http://www.dendroboard.com/forum/food-feeding/67766-repashy-superfly-26.html#post608267)
> 
> ...


 Thanks Ed. I'll have to read that one.


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## Allen Repashy (Jul 17, 2009)

Ed said:


> There is another study using red-eye tree frogs that has some good information in it as well.. Sorry not free access see A brighter future for frogs? The influence of carotenoids on the health, development and reproductive success of the red-eye tree frog - Ogilvy - 2012 - Animal Conservation - Wiley Online Library for the abstract.
> 
> Over the last couple of years I've increased the carotenoid (astaxanthin) content of the Calcium plus formula until it's about 50/50. I've seen better reproduction and an increase in red coloration of the pumilio. (And as a side note, that is with a once/week feeding of D. melanogaster and no other supplementation). I think I've seen all I can figure out using this method so will be switching up my trials in the near future.
> I saw a huge amount of froglets produced out of my caucheros. I think I'm up to nine froglets in the last 4 months from one pair (there is at least one extra in their cage now as I suddenly have two calling males again).
> ...


You can download the referenced article from my website at: http://store.repashy.com/media/Brighter Future for Frogs.pdf


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## ecichlid (Dec 26, 2012)

Thank you JPccusa for pointing out the answer for my first question.

I did not see the answer to my 2nd question. (I'm not entitled to an answer, but I sure would appreciate one so I can make a purchase and have the healthiest frogs possible) I'll ask it another way. *If I have a pound of generic fly food, how many ounces of SuperPig should I include in that pound?* Or if you could tell me how many ounces of SuperPig to add to a pound of fly media. 

Thank you in advance Allen! (or anyone who is better at reading comprehension and can point out the answer to me.)


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## Allen Repashy (Jul 17, 2009)

ecichlid said:


> Thank you JPccusa for pointing out the answer for my first question.
> 
> I did not see the answer to my 2nd question. (I'm not entitled to an answer, but I sure would appreciate one so I can make a purchase and have the healthiest frogs possible) I'll ask it another way. *If I have a pound of generic fly food, how many ounces of SuperPig should I include in that pound?* Or if you could tell me how many ounces of SuperPig to add to a pound of fly media.
> 
> Thank you in advance Allen! (or anyone who is better at reading comprehension and can point out the answer to me.)


This is really impossible to answer because every fly media is different and I can really only speak for my own.

I would say to just use the same levels for any media as using 5% in a media with zero carotenoids will still have more overall carotenoids than the study media.


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## MeiKVR6 (Sep 16, 2008)

For those who raise Pumilio & occasionally offer Bug Burger to the isopod populations within the vivarium directly... Why not mix superpig into bug burger, when it's offered to isopods within the enclosure? Sounds like it could be another more beneficial gutload, maybe...  Any chance that would work, Allen?


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## Allen Repashy (Jul 17, 2009)

MeiKVR6 said:


> For those who raise Pumilio & occasionally offer Bug Burger to the isopod populations within the vivarium directly... Why not mix superpig into bug burger, when it's offered to isopods within the enclosure? Sounds like it could be another more beneficial gutload, maybe...  Any chance that would work, Allen?


You could do that also.... of course we can't assume any similarities between Iso's and Drosophila as far as retention, assimilation, or conversion to an "x" factor.... 

It couldn't hurt though and i would recommend the same inclusion as for the superfly.

Allen


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## hypostatic (Apr 25, 2011)

frogfreak said:


> That wasn't the purpose of the study and would take a new experiment to get the results. I'm not sure that would even be possible...
> 
> I'm just glad they did this study. It's not everyday we get this kind of info! It's groundbreaking really, don't you think?
> 
> Glass half empty kinda guy?


Well.... that's the main problem with scientific papers, there's always more experiments to do and not enough time 




Ed said:


> Over the last couple of years I've increased the carotenoid (astaxanthin) content of the Calcium plus formula until it's about 50/50. I've seen better reproduction and an increase in red coloration of the pumilio. (And as a side note, that is with a once/week feeding of D. melanogaster and no other supplementation). I think I've seen all I can figure out using this method so will be switching up my trials in the near future.
> I saw a huge amount of froglets produced out of my caucheros. I think I'm up to nine froglets in the last 4 months from one pair (there is at least one extra in their cage now as I suddenly have two calling males again).
> 
> Some comments
> ...


Wait, am I understanding that you are mixing equal ratios of Cal+ and carotenoid? Like 1 cup each?


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## Ed (Sep 19, 2004)

hypostatic said:


> Wait, am I understanding that you are mixing equal ratios of Cal+ and carotenoid? Like 1 cup each?


Yes but not on that volume. I only mix as much as I need for approximately two weeks of feedings. So it's more like 1 table spoon of each. It was a trial for the last several years to see if I could find a deep end on the astaxanthin levels. I did not see anything that gave me any indication I had found one. 

I'm now going to change what I've been doing and see what happens.. 

Some comments 

Ed


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## Celtic Aaron (Jun 12, 2013)

Allen Repashy said:


> You could do that also.... of course we can't assume any similarities between Iso's and Drosophila as far as retention, assimilation, or conversion to an "x" factor....
> 
> It couldn't hurt though and i would recommend the same inclusion as for the superfly.
> 
> Allen


MeiKVR6 or Allen,

I realize that I am a little late on this conversation, but I am learning a lot and trying to catch up . With regards to Bug Burger for springs, am I correct in assuming that you just sprinkle some under the leaf litter within the viv itself? Would this have any effect on the springs ability to "clean" the tank? I have just been feeding my spring culture once a week or so and then feeding them off in the viv assuming that they are eating decomposing matter within the viv. I appreciate any help.

Aaron


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## frogparty (Dec 27, 2007)

theres a whole thread about bug burger! check it out



http://www.dendroboard.com/forum/food-feeding/129746-repashy-bug-burger-isopods.html


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## Celtic Aaron (Jun 12, 2013)

frogparty said:


> theres a whole thread about bug burger! check it out
> 
> 
> 
> http://www.dendroboard.com/forum/food-feeding/129746-repashy-bug-burger-isopods.html


frogparty,

Thanks for the thread, I appreciate the help. That is exactly what I was looking for. 

Aaron


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## elianto (Feb 15, 2009)

hey,

sorry for resuming the topic,

i just recently puchased superPIG and i was wondering, should it be directly dusted on the feeders or integrated in the gutloading of fruitflies and collembola?

thanks


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## ZookeeperDoug (Jun 5, 2011)

You can do both.


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