19 replies to this topic

[Etherwulf]

 

 I've finally gotten round to creating a research thread for my synthetic ant diets. I aim to provide some insights into formulating synthetic diets as well as show that it is fairly easy to accomplish.

 

Nonetheless, there are certain pitfalls along the way that I will highlight so that you need not repeat my previous mistakes. As far as possible, each step will be illustrated with a picture but for those that do not, there will be clear descriptions of the steps taken.

 

 

Introduction

 

Synthetic ant diets are nothing new. Several research papers have documented the creation of these diets with the most prominent ones below:

 

Straka J. and Feldhaar H. 2007. Development of a chemically defined diet for ants. Insect. Soc. 54: 100–104 

 

Bhatkar A.P. and Whitcomb W.H. 1970. Artificial diet for rearing various species of ants. Florida Entomol. 53: 229–232

 

Dussutour A. and Simpson S.J. 2008. Description of a simple synthetic diet for studying nutritional responses in ants. Insect. Soc. 55: 329–333 

 

Ants require two main types of food, carbohydrates and protein. Generally, worker ants require very little protein and rely on carbohydrates for energy while larvae require a much larger amount of protein to grow. Also, adult ants can only feed on liquid foods and only larvae can consume both solid and liquid foods.

 

The key to creating successful diets is the protein to carbohydrates ratio (P:C). Several studies have shown that too much protein can lead to lifespans being shortened by half and colonies collapsing. In short, too much protein will kill your ants so great care must be taken in deciding the amount of protein. 

 

The Dussutour and Simpson diet (Which is what Formula is based on) found that the most successful combination for Rhydontiponera ants is a P:C ratio of 2:1 so that is what we will be using later. However, I have made modifications to the diet which have given it greater receptivity to certain species of ants that previously refused to recruit to the original. Also, egg has been omitted from the diet because a research done on egg white injury in ants have provided sufficient reason to remove it. The study showed that ants fed on  egg white diets had shortened lifespans and showed increased aggression towards nestmates. 

 

Creating synthetic ant diets offer a more consistent source of nutrition and is also easier to feed in the long run. In addition, it is more economical because very little of each ingredient is needed in making one batch.

 

Ingredients 

 

Protein

 

For protein, there are a few choices to pick from:

 

Egg

 

Whole eggs are a high quality source of protein, or more specifically, albumin. It has been used in ant formulations since the original Bhatkar-Whitcomb diet but it is known to cause egg white injury or biotin deficiency when used in high quantities. Hence, I have chosen to omit it from my own formulations. If you choose to use eggs as a protein source, you must ensure it is used in conjunction with other protein sources.

 

Calcium caseinate

 

This is normally found in milk alongside whey. However, it is known to sediment in solutions as it is the least water soluble of the caseinates, causing an uneven distribution of protein. Hence, it is advised to avoid calcium caseinate.

 

Whey concentrate

 

Whey is also found in milk and is available as a concentrate or isolate. It is commonly consumed by bodybuilders as protein so it can purchased from your local health store. Isolate contains a higher protein percentage of about 90 - 94% while concentrate has 70%-80%

 

Insect Extract

 

Insect extracts are perhaps the best source of protein since they are low fat (with the exception of waxworms) and are consumed naturally by ants. I prefer cricket flour which can be bought online or from a health store. Make sure you buy only those with no preservatives and no additives.

 

Carbohydrates

 

Fructose and Sucrose

 

Fructose and sucrose are found in honey and table sugar. Fructose is slightly sweeter than sucrose. Honey is prefered here because it is minimally processed so it keeps all trace minerals and vitamins, instead of refined sugar which are "empty calories". If you can obtain unrefined sugar, that would be best but it can be somewhat tricky to get hold of.

 

Preservatives

 

Sodium Benzoate

 

Sodium benzoate is used to as an antimicrobial agent to inhibit mold growth. It is easily obtained at stores selling baking supplies. It is only effective at a pH of below 4.5 so ascorbic acid (vitamin C) needs to be added to reduce the pH. If you choose to use vitamin C tablets, ensure that no flavourings and additives are added.

 

Vitamins/Others

 

Vanderzant Vitamin

 

This is specially designed for insects but can be hard to obtain. Use sparingly (about 2g per 300ml).

 

Agar

 

Used to solidify food into cubes or gel

 

Ascorbic Acid (Vitamin C)

 

Used to increase the acidity of the solution to increase the effectiveness of sodium benzoate at inhibiting mold growth

 

Food Coloring of choice

 

Tools

 

 

Milligram scale

 

This is used to measure precise quantities of ingredients. I use a cheap diamond scale bought online.

 

Stove/ Microwave

 

I prefer using a gas stove because it offers me greater control over the heat. A microwave can be used but a bit of finesse is required because the solution should be taken out when it starts to boil. 

 

Spoons/Stirring rod/Beakers

 

Small coffee spoons can be used to scoop minute amounts of powder ingredients.

 

pH scale is not needed but can be used to double-check the pH.

 

Steps

 

 

1. Ensure environment and tools are clean

 

Wash all tools with soap and sterilize metal/glass tools in boiling water if possible. This is to maximise effectiveness of the preservative used. Wash hands thoroughly with soap before beginning.

 

Common mistake: Do not skip this step! Skipping this step can cause mold to grow despite the presence of sodium benzoate. Observe the picture below for details.

 

(Note that the bottom left was from a contaminated batch that still had preservatives. These samples were left for a week in a moist environment to simulate the likely conditions that your ants will be living in.)

 

 

2.  Prepare the following quantities of ingredients:

 

4.916g (3.335g)* of cricket powder 

2.222g (1.665g)* of whey 

12.220g (10g)* of unrefined honey (Percentage of sugar varies based on honey used. Check nutrition labels)

100ml of distilled water

1.300g of agar (Increase or reduce this to change consistency of the final product. I used a 1.3 % agar solution at a 5 :1 ratio of agar solution to dry mass of ingredients)

0.120g of sodium benzoate

2.000g of ascorbic acid (You may have to increase this if you use more water) 

1g of Vanderzant 

2 drops of food colouring

 

 

*brackets indicate actual amounts of protein or sugar present

 

Common mistake: Do not approximate! Weigh all ingredients using the milligram scale. Small deviations of ±0.01 are acceptable.

 

Tip: You can weigh certain ingredients together to save time. Use a small container/dish to hold the ingredient being weighed but remember to 'tare' the scale to 0 before adding the ingredient.

 

 

 

        

 

3. Dissolve all ingredients EXCEPT AGAR in 40 ml of distilled water

 

Stir until all ingredients are dissolved. 

 

Tip: The honey is likely to stick to the weighing container. Rinse with water and pour into the mixture until no more honey is left. Better still, use a larger container to weigh the honey and just mix ingredients in that container later.

 

 

 

4. Dissolve agar strips in boiling water

 

Boil 100ml of distilled water before adding agar strips. Stir until dissolved and switch to low heat

 

5. Add the mixture from step 3 to the agar solution on low heat

 

Stir until the solution is homogenous and evenly spread out. 

 

Common mistake: Do not allow parts of the solution to agglutinate/clump up! This will cause an uneven distribution of protein and carbohydrates

 

 

*OPTIONAL STEP*  Skip to step 6 if you don't want to pasteurize your ant diet. Pasteurization kills microbes and allows your ant food to be stable-shelf. That said, refrigerating it is still prefered.

 

5a. Heat solution at a constant temperature of about 75 celsius for at least 20 seconds before cooling it to 4 Celsius in an ice bath or fridge.

 

6. Pour mixture into a shallow dish or mold (if you want shapes)

 

I used sterile petri-dishes but you can use any shallow dish that has been cleaned with soap or sterilized.

 

Common mistake: Ensure dish or mold is cleaned with soap or sterilized. Do not touch the mixture with your hands!

 

 

 

 

7. Refrigerate completed ant diet at around 4 Celsius 

 

To store it long-term, put it in the freezer compartment.

 If you have pasteurized your food in step 5a, it is fine to store it at room temperature. 

 

 

Whew! If you have followed the steps above, congratulations, you have just made your first batch of ant food. 

 

This is my first version using cricket flour and I'm already seeing greater receptivity. Picky species like Nylanderia sp. readily accepted this version so if you have trouble feeding your ants, try this out.

 

Feel free to modify this formulation and post your own attempts below. 

 

If enough people are interested, I will create an online logsheet to allow everyone to share their own synthetic ant diets.

 

FAQ

 

Can I feed my ants solely this?

 

While studies have shown that healthy colonies have been raised solely on this, I recommend using this as the staple food while offering the occasional insect. Water should be provided at all times. If your colony does not grow as expected, cease feeding a nd provide sugar water ad libitum along with insects 

 

Feedback this to me on this thread so I can figure out what's wrong

 

Isn't this the same as Formula by Byformica?

 

No.

 

While Terry and I base our diets off mostly the same literature, we have made our own modifications to the original D&S diet so they are very different in composition.

 

The PC ratio for this particular diet remains the same but I've increased the amount of sugar for sugar-loving species which refused to take the P:C 2:1.

 

This sounds like a lot of work. Can I buy some from you?

 

Currently, I have no plans to sell this. I am planning on sending free samples in the future to people in exchange for them recording their observations for me so I can improve this diet. If there is enough interest and when I have time of course, so drop me a PM to indicate your interest.

 

My ants don't want to eat this! What's wrong?

 

Again, many factors are involved such as consistency, preservative levels, etc. Try increasing amounts of honey used and remove other competing sources of food.

 

Feedback this to me on this thread so I can figure out what's wrong

 

Can I use a liquid feeder with this?

 

Yes but I do not recommend it. Adding more water will result in a gel but it will eventually agglutinate, causing the feeder to clog.

 

Can I repost this somewhere else?

 

Yes, but credit me and provide a link back to this page.

 

 

Moderators, please sticky this if you find it beneficial.

 

And for all those who've found this helpful, please like this post. Thanks! 

 

 

Edited by Etherwulf, September 22 2015 - 6:14 AM.

[C Carl]

Very informative post, Etherwulf.
 
I have used a variation on the Dussutour & Simpson diet with some success, but may incorporate some of your modifications.

 

I find it easier to pour the diet into small loaf pans, using portions of a light fixture diffuser as a dividing grid. Once solid, the uniform chunks can be scraped into clean petri dishes for storage. I do have access to a clean transfer hood for this part, so the extra handling isn't introducing contaminants.

 

Attached Images

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[dspdrew]

Very nice thread.

[Trailandstreet]

What disturbes me most is, that one can buy cricket flour as health food.

[Crystals]

I have added it to the List of Handy Links thread which is stickied in the General section.

[Foogoo]

 Also, adult ants can only feed on liquid foods and only larvae can consume both solid and liquid foods.

Can somebody elaborate on this? I've heard it multiple times, but then how do you explain queens eating eggs? I'm assuming workers "eating" solid foods are taking it to feed larvae and not eating it themselves?

 

What disturbes me most is, that one can buy cricket flour as health food.

I got a box of ranch flavored roasted crickets at a candy store once, surprisingly good!

[dean_k]

Ants cannot eat solid food at all. Larvae can eat solid food.

 

Eggs are basically liquid.

[Foogoo]

Ants cannot eat solid food at all. Larvae can eat solid food.

 

Eggs are basically liquid.

Same with the ants that crawl all over the cookies, chocolate and sugar you left uncovered. All of that is for the larvae?

[dean_k]

Yes, ants cannot simply digest dried food. They can't even eat (or swallow) it. When they bring in solid food, it's for larvae.

[dspdrew]

 

 Also, adult ants can only feed on liquid foods and only larvae can consume both solid and liquid foods.

Can somebody elaborate on this? I've heard it multiple times, but then how do you explain queens eating eggs? I'm assuming workers "eating" solid foods are taking it to feed larvae and not eating it themselves?

 

This is confusing to me as well. I have spent a long time watching my ants through my microscope, and probably 99 percent of the time they will suck all the juice out of the egg, and then ball up the skin and give it to their larvae. But, there have been a few times when I watched them with my own eyes, gobble up the entire egg. It all went in their mouth, and never came back out, so I have no idea what that was all about. I've seen them do that with small pieces of insects too.

[dean_k]

I believe ant's digestion system is similar, if not the same, with spider's digestion system.

 

Their diet must be something of high moisture content. Liquid form is ideal. If not, moist solid form is possible to be chewed and inserted into their stomach. In case of consuming eggs, egg shells are soft and moist. Thus, they should be able to eat it if they chew it into smaller pieces. But it's not their ideal way of consuming food.

[SMILEforAnts]

Cool nice recipe! Very detailed.

 

I didn't even know cricket flour existed. I looked it up and it says you can use it in food recipes, especially baking. o_O It is true when they say we'll be using more insect protein in the future.

[Etherwulf]

Very informative post, Etherwulf.
 
I have used a variation on the Dussutour & Simpson diet with some success, but may incorporate some of your modifications.

 

I find it easier to pour the diet into small loaf pans, using portions of a light fixture diffuser as a dividing grid. Once solid, the uniform chunks can be scraped into clean petri dishes for storage. I do have access to a clean transfer hood for this part, so the extra handling isn't introducing contaminants.

 

That's great to hear Carl. Could you share with us the variations that you have tried as well as the results/effectiveness?

 

The formula I showed is what has been effective for most of my colonies, even the picky ones. Also, growth rate is on par with colonies fed insects and sugar with no distinct difference in worker mass, pupae mass or brood production.

 

Take note that 'combined' diets may not offer the right balance of protein and carbohydrates to maximise colony growth for certain species like Solenopsis. The requirements of colonies on a nutritional level also varies based on several factors such as the number of larvae. Hence, if you are not achieving the expected growth rate, you may wish separate protein and carbohydrates into individual cubes. That way, the ants can choose whether they need protein or sugar. A combined diet that performs well for all species is difficult to attain so a bit of tweaking and experimenting is needed for each species.

 

Also, I hear you are studying invasive species (e.g Nylanderia fulva). It would be nice to share your findings with us  if you can.    

Edited by Etherwulf, September 22 2015 - 6:13 AM.

[C Carl]

For my Nylanderia fulva, I increased the carbs by adding sucrose such that the final ratio is 1:4.5, instead of 1:2, of protein to carbohyrates. My reasoning for the change goes back to this specialization of digestive capability for the different stages. I was keeping only workers that have little ability to digest protein. My understanding is that a normal colony truly is acting as a super-organism in this respect, requiring the contributions of the immatures to provide the stream of protein for the rest of the nestmates. I don't have statistical evidence to back it up, but the workers seemed to be visiting the artificial diet more frequently with the sweeter blend. I also provided the ants with a 10% honey water solution. Hopefully, though, some of the protein in the diet blend was still being assimilated. The Dussutour and Simpson diet I modeled mine after uses water, agar, whole egg powder, whey protein, calcium caseinate, Vanderzant vitamin mix, sucrose and methyl paraben for a preservative. Obviously, this is not a "colony" and has no growth rate. Especially when kept at cooler temperaures, I would have good survivorship well past 6 months for these workers, so maybe something good was happening. The calcium caseinate is indeed a bear to get into solution.

 

I like your suggestion of separating the components to allow choice for selective foraging.

 

I'll have to get back later about the research findings as we are still in the early stages.

[Foogoo]

Relevant for curious minds:

 

Laboratory fire ant colonies (Solenopsis invicta) fail to grow with Bhatkar diet and three other artificial diets J. Gavilanez-Slone • S. D. Porter

 

Why do we otherwise have to pay $40 to read a paper written and based off research by the USDA? Same with research papers from other government agencies and those funded by public grants. The system is broken...

Attached Files

•  ants.pdf   242.61KB   161 downloads

[Etherwulf]

Yes, Foogoo, I've previously discussed that study with Terry as well. 

 

The conclusion that we came to was well, inconclusive. This is largely because there is a lack of sufficient testing across different species and the reasons the authors postulated to have caused the results in the the Dussutour-Simpson study as well as their own.

 

They hypothesised that it was the suboptimal temperature that the study might have been conducted at which would result in a slowing of growth rate. However, isn't it strange that that the colony fed on the synthetic diet grew at roughly the same rate as the colony fed on fruit flies and sugar? Assuming that temperature did cause a decrease in growth rate, one would expect the synthetic diet colony to fare far worse than the fruitfly/sugar colony and this was clearly not the case. The difference was in actual fact, marginal.

 

Seeing that they tried to extrapolate from their results on Solenopsis invicta and suggested unlikely reasons for the relative success of the Dussutour-Simpson study, I would pause before coming to any conclusions about synthetic diets or using this study as a basis for arguing that synthetic diets are not suitable for rearing a healthy colony. Maybe a Solenopsis colony but other species? Not so much.

 

So far, the development of synthetic diets is still in its infancy because of a lack of comprehensive testing across a wide range of species with empirical data. Most of the studies that did utilise synthetic diets did not mention growth rate as they were basing their expected results off D&S and were investigating something else anyway. Hence, I have repeatedly discussed the possibility of conducting a study of "separated diets" vs "combined diets" on different species with  sufficient sample sizes.

 

However, there are multiple obstacles such as the complexity of the study in question. It would require a large amount of time and access to resources that I do not possess so I don't see myself conducting it anytime soon.

[Gregory2455]

That pink one looks pretty funny. That is bacteria???

[panda0224]

Would you recommend to use autoclave for sterilization or do you think it would break down the ingredients to the level they are no longer beneficial to ants? 

It would be around 122C at 18 psi for ~20 minutes

[benjiwuf]

You'll be burning off most of the vitamins and denatured the proteins, but the rest will be fine.

Edit: I would also add the ascorbic acid and benzoate after it cools down a bit also. Acids tend to also break down with heat.

Edited by benjiwuf, March 23 2017 - 9:35 PM.

[KBant]

Thanks for posting forumila!! Very detailed!