# new member, FF culture expert



## Guest (Aug 6, 2005)

just wanted to introduce myself.. recently got interested in PDFs when i found out that ppl actually have em as pets. been reading this forum for a few days now and am thinking of getting some leucs as my first froggies for a 20gal viv.

anyway, if ya'll have questions regarding FF, i'm pretty much an expert, i work with em all day at a developmental genetics research lab so i can give you the lowdown on culture techniques and possibly add some new knowledge to the board.

DR


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## amphibianfreak (Jul 21, 2004)

Welcome to the board.


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## dmatychuk (Apr 20, 2005)

EXCELLENT and welcome. Please begin the brain dump. Don't wait for someone to ask the right question because we might not even know what to ask and you might mention something that has not been addressed here on the board. My first topic of interest would be that I want to make my FF as potent as possible for my frogs so:
-What is the best specie?
-What is the best food?
-Is Brewers yeast helpful health wise?
-Will adding Whey to the culture help protein levels in the FF or is there something that will?
-What is the best way to enrich the FF with calcium internally?
-Has real research been done on protein, calcium and fat levels of FF, and how to effect it? If yes what are the results?

I don't want a discussion on dusting but what can be done with the ff itself.


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## Guest (Aug 6, 2005)

dmatychuk said:


> EXCELLENT and welcome. Please begin the brain dump. Don't wait for someone to ask the right question because we might not even know what to ask and you might mention something that has not been addressed here on the board. My first topic of interest would be that I want to make my FF as potent as possible for my frogs so:
> -What is the best specie?
> -What is the best food?
> -Is Brewers yeast helpful health wise?
> ...


i've noticed that the main hurdle when it comes to PDFs seems to be the food source and its proper raising and nutrient profile, and as such i've started a few little of my own side experiments at my lab to answer some of the questions you just asked me. i work with d. melanogaster which i think would be the best due to fastest life cycle. 

we actually use brewers yeast in our cultures... let me describe them really quickly. we use little plastic flasks filled with molasses as the sole ingredient, and we springkle a tiny bit of dry yeast on top of this medium (not too much or the yeast will take over the jar). this is mainly for the larvae, which love it. they grow like crazy in this environment at room temperature, if you want them to grow even faster though keeping them at 25C will speed up their life cycle. with this food method they get everything they need, the cultures never go "bad" unless you happen to not get a female in one of them and then no eggs are laid. the cultures don't get infected with mold/etc because the molasses has been boiled for pouring into the flasks.

as for the actual nutrient profile of the flies, there are no studies out there on it, but i invented my own little "experiment" to see if there could be a better method of loading the FFs with more nutrients other than dusting, which seems like a pain and highly inefficient. so what i set up is a little experiment in which i "starved" the flies for 1-2 days by placing them in foodless flasks (with a slightly damp paper towel in there for moisture), we use this same method when we want to feed the flies mutagens, after starving they will eat anything. to fatten them up for ovary dissections, we feed them 5mls water + 3g yeast that has been thoroughly mixed together into peanut butter consistency, they LOVE this and will gorge themselves on it... so i did the same for this experiment except i added red dye so that i could track how quickly they feed and how long it takes their abdomen to become completely filled with the food. turns out within 30 minutes most of them have eaten enough to become almost fully red, by about 1-2hrs it's safe to say that they have become engorged with red dye. so, knowing this, it's not that hard to figure out that if you mix in herp vitamins/calcium (and other goodies that ppl seem to add like beet powder or paprika, even mixing in whey for protein would work) into the yeast "butter" instead of the dye, within an hour of feeding after a day of starvation they will be LOADED with this good stuff. 

i hope this all makes sense, feel free to ask if something is unclear, but i think this little method i came up with could get rid altogether of the dusting business.


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## dmatychuk (Apr 20, 2005)

EXCELLENT information. Any chance you could get pictures of any or all of this process. I would like to see the flasks full of your mix and the yeast paste and if possible a loaded ff. When you say yeast for the paste are you using brewers?


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## Guest (Aug 6, 2005)

dmatychuk said:


> EXCELLENT information. Any chance you could get pictures of any or all of this process. I would like to see the flasks full of your mix and the yeast paste and if possible a loaded ff. When you say yeast for the paste are you using brewers?


we use normal grocery-store bought yeast, but i'll check on the exact kind on monday. i don't have a digi cam but i'm thinking of getting a cameraphone soon so i'll take pics then and post up.


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## Jace King (May 5, 2004)

Well you obviously know more about Fruit Flies than me, but I dont think i would ever be cormfortable giving up my repcal and herptivite.


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## slaytonp (Nov 14, 2004)

Oh my, you are really asking for it!! :lol: Be prepared for questions and welcome aboard.


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## Alfy111 (Aug 6, 2005)

So wait, you use molasses with some brewers yeast sprinkled on top? Thats so simple!  And then to put in the nutrients you mix in some vitamin supplements with the food, but first you have to starve them for a few days? I just want to make sure Im getting this right...


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## Guest (Aug 7, 2005)

Wow, this is really awesome and welcom to the addiction.

This, only after reading, seemed very much like common sence. But until I read it here was never thought of, at least by me.
That is why I love these boards.
On the maple syrup, did you get it from the store and then boil it again? I already have the syrup since I hate the flavor of the fake syrups myself, to each his own.
So, If I read correctly you boiled it again added to a beaker and let it cool. Added a few grains of yeast and some flies to the culture, and voila?
What type of stopper do you use in the lab. I have a scientific supply store close to me and could easily get the same stuff.
This would be great, never loose the culture and easy to take care of, no more 20 different cups, only 10 for rotation!!!!
OH another thing, using molassas in a beaker, how do you extract the flies without slopping the molassas on the sides of the beaker?

Thanks for the clarification and thank you for you expertise, it will help raise healthier froggies in the future.


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## Guest (Aug 7, 2005)

Alfy111 said:


> So wait, you use molasses with some brewers yeast sprinkled on top? Thats so simple!  And then to put in the nutrients you mix in some vitamin supplements with the food, but first you have to starve them for a few days? I just want to make sure Im getting this right...


you, my friend, get an A+ in reading comprehension =) you got it


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## Guest (Aug 7, 2005)

fletch33 said:


> Wow, this is really awesome and welcom to the addiction.
> 
> This, only after reading, seemed very much like common sence. But until I read it here was never thought of, at least by me.
> That is why I love these boards.
> ...


it's not a "syrup," it's a VERY thick molasses that solidifies and doesnt move at all. i will ask the person who prepares the food at work on monday exactly what she uses and how she does it. 

these are the plugs we use w/ our disposable plastic bottles, they arent cotton so mites can't get in


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## Guest (Aug 7, 2005)

Could you please post the maker or better yet a picture or link to these containers?
I was thinking of using a filtering flask with a 1/4" tube to a collection jug. IT this jug I could have the peanut butter paste that you mention. Maybe have 3 flasks feeding this one jug and that way I could feed from this one jug that is pre gut loaded, daily.
Have 2 of thses setup in case a batch goes bad, well with molassas the only problem would be contamination from an outsider.
In the air line I could use a 1/4" air valve for when ready to feed flip the valve so no others could escape.
What do you think?


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## Guest (Aug 7, 2005)

fletch33 said:


> Could you please post the maker or better yet a picture or link to these containers?
> I was thinking of using a filtering flask with a 1/4" tube to a collection jug. IT this jug I could have the peanut butter paste that you mention. Maybe have 3 flasks feeding this one jug and that way I could feed from this one jug that is pre gut loaded, daily.
> Have 2 of thses setup in case a batch goes bad, well with molassas the only problem would be contamination from an outsider.
> In the air line I could use a 1/4" air valve for when ready to feed flip the valve so no others could escape.
> What do you think?


sounds good to me, i JUST ordered a cameraphone online so in a few days i'll hopefully be able to post pics of exact setup and i'll write up a clear step-by-step illustrated procedure. it's basically the same exact protocol every drosophila lab uses and pretty much failproof, so i think this should make the PDF hobby a lot easier once you guys know how to do it.

i also have some other ideas concerning gut-loading the flies that i will investigate and post about later.


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## Guest (Aug 7, 2005)

You are making everyone's life so much easier!! Im glad you came along.
I just got a job as a College lab tech ( i start tomorrow) and who knows, I might have to make some fruit fly cultures some day.


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## Michael Shrom (May 20, 2004)

Your technique sounds interesting. I like to use a medium that does not need to be boiled or cooked. I prefer something you can mix and add flies to immediately. I also need large hatches that can be harvested with as little time invested as possible. Anybody that has kept frogs for any length of time can tell you that the food is the hardest part.


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## Michael Shrom (May 20, 2004)

Your technique sounds interesting. I like to use a medium that does not need to be boiled or cooked. I prefer something you can mix and add flies to immediately. I also need large hatches that can be harvested with as little time invested as possible. Anybody that has kept frogs for any length of time can tell you that the food is the hardest part.


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## Ed (Sep 19, 2004)

There have been nutritional analysis of D. melanogaster and they have been published one such is listed below (and expect a more complete analysis to be published in the new edition of Mader's book). 

I refer you to Bernard, Joni B; Allen, Mary E.; Ullrey, Duane E.; 1997, Feeding Captive Insectiverous Animals: Nutritional Aspects of Insects as Food, AZA Nutritional Advisory Group Handbook (data adapted in part from Allen, M.E.; 1989, Nutritional Aspects of insectivory; doctoral dissertation, Michigan State University, East Lansing, MI.)


D. melanogaster

adults larva pupae

dry matter 29.6% 21.2% 32.4%

crude protien 70.1% 40.3% 29.6%

crude fat 12.6% 29.4% 10.5%

total minerals 4.5% 9.8% 14.1%

ADF 27% 5.9% 17.4%
(acid dry fiber)

gross energy 5.12% 5.57% 4.84% 
(kcal/gram)


There will some overlap as crude protien will include chitin as will ADF. (so if you add the total up it will be more than 100%)


major and trace mineral contents on a dry matter basis


adult larva pupae

Ca 0.1% 0.59% 0.77%
P 1.05% 2.3% 2.73%
Mg 0.08% 1.89% 2.41%
Na 0.42% 0.09% 0.12%
K 1.06% 1.28% 1.66%

Cu 18 ppm 16 ppm 25 ppm
Fe 138 ppm 235 ppm 1728 ppm
Zn 171 ppm 176 ppm 200 ppm
Mn 39 ppm 110 ppm 108 ppm
Se 0.07 ppm 0.49 ppm 0.33 ppm



As can be seen by the ratios above the calcium to phosphorus ratios are very imbalanced (as a good ratio would be between 1:1 and 2:1). 

Starvation of the insects has been shown to deplete fats, protiens, and trace elements (See the nutrition chapter in Mader's book, Reptile Medicine and Surgery) that can take at least 24 hours and up to 72 hours to replenish. If this example continues to hold true with flies (and I have seen nothing to indicate that this truism would not hold true given that it also applies to more than one genera). In the species that have been studied to date, gut loading is insufficient to make up the difference in the nutritional content due to starvation. 

Some comments,

Ed


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## dmatychuk (Apr 20, 2005)

ED,
That is great information! Do you happen to have a link to anything like this? I wonder if rather than starving and gut loading if one were to 24 hours prior to feeding to their PDF they only give the FF food that is high concentrate of protein, calcium and vitamins?


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## Ed (Sep 19, 2004)

If I get a chance to write up the powerpoint talk I gave at IAD, I have at least on request to allow it to be put on a website. It will contain some of this information. 

This NAG publication was not online when I checked the website but is part of the publications published by the NAG. 
Ed


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## Guest (Aug 8, 2005)

This is why I was thinking of making a flask setup with a central container with the paste. The flys would crawl throu the 1/4" tube to the center beeker or container and gorge there on the past. This way they follow the stink and I only would feed the flys from this one container, non starved and gut loaded.
Unless you say that they will not feed a few hours after hatching?
I will have to get a few flasks and a new culture of flies.

This is becoming a very informative post.


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## Ed (Sep 19, 2004)

Off the top of my head I am unsure whether or not the flies feed within a couple of hours post completion of pupation. 

However there needs to be some discussion of "gutloading" here. Gutloading was originally the phrase coined by a group of researchers who were attempting to address the imbalance between calcium and phosphorus in feeder insects by manipulating the diet of the feeder insects through high calcium diets. High calcium diets can work but only in specific circumstances as they are unpalatable to the insect, significantly increase the mortality of the insects that they are being fed to, do not work in all instars of the insects studied (for example, in crickets, only pinheads acheived a positive calcium to phosphorus ratio) and need to be fed for a significat period of time for the calcium to accumulate in the digestive tract (temperature dependent but usually a minimum of 48 hours (with mortality spiking about 72 hours)), need to be the sole diet offered for a significant period of time, show a similar reduction in calcium content over time when compared to dusting and do not address the dietary requirements of the frog with respect to D3 as the anuran (or other animal) may be unable to metabolize the calcium when fed via gut loading. 
At this time, to the vast majority of hobbyists, gutloading is simply applied to any diet fed to feeder insects that is percieved to modify the nutrition of the feeder insects. With a lot of fat stored nutrients (such as carotenoids), it is probabaly sufficient to simply include it in the media supplement with the flies as the adults will also feed on the media which includes the supplement. 

Some comments,

Ed


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## Alfy111 (Aug 6, 2005)

Alfy111 wrote: 
So wait, you use molasses with some brewers yeast sprinkled on top? Thats so simple! And then to put in the nutrients you mix in some vitamin supplements with the food, but first you have to starve them for a few days? I just want to make sure Im getting this right... 


DRosophila wrote: you, my friend, get an A+ in reading comprehension =) you got it

haha thanks.. that made my day lol.. hopefully that my reading comprehension will hold up in highschool(freshman).. lol :wink:


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## Guest (Aug 9, 2005)

ed, i like your "comments," you bring up very good points. i will keep tinkering with the flies at work and maybe send some off for analysis. will also look into a few of the points you brought up and then report back.

for the others, pics of the setup will come as soon as i receive my phone. FF culture need not be complicated or have any noticeable failure rate.


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## reptileink (May 1, 2005)

How hard is it to boil the molasses, or should I say, how messy?? I have had to resort to pinhead crickets the past month due to my reptile shop not making many cultures as of late. I would like to make my own, and your method sounds uber easy. Thanks!


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## JoshKaptur (Feb 17, 2004)

My understanding based on the early comments was that is was molassas plus bakers yeast, not brewers yeast. So Alphy, one of us has a reading comprehension problem (I hope it's me so your day is not un-made)... please clarify DRosopila. The "supermarket" yeast is almost always bakers yeast... and adding just a pinch of breweres yeast would do pretty much nothing for a culture (it certainly would provide no active yeast).

Josh


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## Guest (Aug 9, 2005)

just checked and we use "red star active dry yeast"

no clue if it's brewer's or bakers to tell you the truth, i'm not the person who makes the fly food but i will be talking to that person shortly.


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## Ed (Sep 19, 2004)

I don't have the reference but one of the limiting factors in production of ff colonies is protien content in the media. 
Typically brewers' yeast (such as people buy in bulk) is dead and adds protien to the media mix. The addition of the baking yeast is to add a live yeast culture which as I understand it produces the signals to the flies that this is a good food source (such as acetic acid and small amounts of ethanol). 

Ed


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## Arklier (Mar 1, 2004)

DRosophila said:


> just checked and we use "red star active dry yeast"
> 
> no clue if it's brewer's or bakers to tell you the truth, i'm not the person who makes the fly food but i will be talking to that person shortly.


If it's active dry yeast then it's baker's yeast. Active means it's alive.


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## Will (Feb 15, 2004)

DRosophila said:


> just wanted to introduce myself.. recently got interested in PDFs when i found out that ppl actually have em as pets. been reading this forum for a few days now and am thinking of getting some leucs as my first froggies for a 20gal viv.
> 
> anyway, if ya'll have questions regarding FF, i'm pretty much an expert, i work with em all day at a developmental genetics research lab so i can give you the lowdown on culture techniques and possibly add some new knowledge to the board.
> 
> DR


Part of the reason I decided to keep darts was that I had a constant supply of feeders through my research (I also use drosophila as a model organism). If you are referring to standard drosophila media, it is a lot more then molasses and yeast. The basics are water, molasses, yeast (bakers or active) cornmeal and agar. I have yet to hear of a lab that uses straight molasses with sprinkled yeast, seems like a death trap to me. 

Re: gut loading… while it is an ok idea, straight dusting is easier and more reliable. Cycling through a starvation / gut load period before every feeding will become very labor intensive. It is also possible that flies which have ‘cleaned’ their dust of in the vivarium would have an equal gut load from cleaning as starved flies eating a yeast paste nutrient mix. 

Just some thoughts… I’m sure this board will appreciate the tips you have to offer.


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