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The Leptothorax Experiment

lepthorax small ant tiny ant polydomy polygyny

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#1 Offline Karma - Posted July 15 2023 - 12:02 AM


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FYI: For those that just like to skim these posts and focus on the pictures, the "TLDR" or essential parts of these updates will be bolded so you don't have to read the entire thing.




This journal will document my raising and merging of several Leptothorax canadensis colonies in an attempt to create as big a colony as possible. It is understood that many Leptothorax and Temnothorax ants utilize a polydomy nesting strategy, forming several satellite nests in a specific area. Once they are of sufficient size (10-30 workers), I will transfer each of the colonies from their test tube to a nest to try and simulate this behaviour. It is documented in several research papers that Leptothorax/Temnothorax behaviour can vary significantly even within a species depending on where the species is located and its nest/resource availability. I am hoping that I will be able to merge the colonies with ease and have them form several nests in a given area but we shall see. It is possible that this will not go well and the colonies end up killing each other instead of merging.


The nest I plan on eventually moving them into is a large custom THA inception chamber with four entrances that split into eight seperate nests or two small inception chambers each with two entrances that split into four seperate nests. Once I am closer to transferring them I will decide on a design, but this won't happen until next year at the earliest.




On to the ants, I currently have nine queens making up seven colonies, with two colonies having two queens. Five of the six queens were only recently caught over the past few days and have not laid any eggs yet. The other queen I caught in late May and currently has 2 larvae close to pupating, one smaller larvae, and one egg that has begun hatching into a larvae. Leptothorax have two great catching seasons: 1) When queens that mated prior to winter but rejoined their colony (for a greater chance of surviving the winter) leave their original colony in the spring and try and form their own colony (either on their own or through fission) and 2) When they have their actual nuptial flight which often happens in July.


Additional Information:


Leptothorax canadensis queens are dimorphic, with microgynes and macrogynes. Leptothorax microgynes often return to their original colony after mating and are unable to fly far from the nest to mate due to their small size and weaker wing muscles. They also often have less reproductive potential and are less dominant. Leptothorax macrogynes on the other hand are larger, more dominant, and often have greater reproductive potential. Macrogynes more commonly leave their original nest to start a new colony after nuptial flights.


In the queens I have caught there appears to be fairly significant size variation. I have yet to take any specific measurments but the queens appear to range in size from ~3mm to ~4mm. Given this size difference I assume that at least two of the queens I have caught are microgynes, with the rest possibly being macrogynes. This may not be the case, but until they have brood and I am able to compare how many eggs they are able to lay and raise I am going to assume the above is accurate.


Leptothorax/Temnothorax colonies often consist of no more than 100-200 ants depending on the species. For Leptothorax canadensis specifically, it is not uncommon to see most colonies only having 30-70 workers depending on nest/resource availability. With that said, in rare instances wild colonies have been observed to have 300-400 workers in certain Leptothorax/Temnothorax species. Additionally certain species have been able to reach 900+ workers in stable lab conditions, I hope to accomplish something similar.


Feeding Regime:


These ants are semi-claustral and need to be fed during the founding stage. For their first feeding, I gave them each some honey and a small piece of cooked chicken. For future feedings I will be giving honey and a single fruit fly and I plan on doing this once or twice a week. Feeding them will give me a chance to take photos which I will post here along with updates on how the colonies are progressing. I will try and update this journal some time next week with better photos that hopefully include some new eggs. I likely will not have to change this feeding regime until spring of next year.


For now I just have a few poor quality photos of the queen I caught back in late May:




Edited by Karma, February 5 2024 - 5:38 PM.

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#2 Offline Karma - Posted July 20 2023 - 10:24 AM


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Update July 20, 2023:


In the five days since the initial post one of the queens have laid eggs. Some of the queens gasters have grown in size which is a positive sign. In my experience it can take several weeks before a queen lays eggs so I expect the rest of the queens to lay over the next two weeks if they are fertile. I took the photos below during a feeding where I gave each colony some honey and a fruit fly. The pictures were a little difficult due to these ants being so small, should be able to get some better photos next time. I also should have mentioned this in the first post, but after catching these queens I tried to pair them up into 3's or 2's as Leptothorax have a much better founding success rate through pleometrosis. After several hours and several tried combinations, only four queens were cooperative which I decided to split into two colonies each with two queens. All the other queens were either too anxious, aggressive, or hyper aggressive towards other queens. Normally when introducing queens to each other they'll do a antenna boxing max and raise their gasters up in the air (I call this the "dance"). After this "dance", If all goes well they both calm down, they will begin to tolerate each other's presence and interact as if they were two workers from the same colony. The anxious queens, rather than engaging in this behaviour, repeatedly get scared and run in a different direciton. In my experience even if these queens aren't aggressive and don't get aggressed on themselves, they will struggle to lay eggs in the presence of the other queen(s), which is why I remove them. The aggressive queens repeatedly perform this "dance" but will also nip or pull on the other queen. I categorized a queen as aggressive if they continued this behaviour after several initial "dance" interactions. While these queens may still be able to engage in pleometrosis, they are often more dominant which makes them a higher risk for killing other queens once workers arrive. The hyper aggressive queens skip this "dance" and go straight to biting and stinging.


Lastly. to make the updates easier, I'll categorize each colony as A, B, C, D, etc.




Colony A: This is the original queen that I caught back in May from the pictures above. This queen is doing great, as you can see from the photos, her gaster is quite enlarged. Even before her physogastrism, she was one of the largest, if not the largest, queens I had caught. Right now I would put her length at around 4mm or more. She has two large and two small larvae which are continuing to grow and I expect will workers by October. The second photo shows the two smaller larvae in better detail as they are so close together it can look like a single larvae from far away. Next to the larvae is a piece of chicken which the queen has been feeding the larvae. Also to clarify, for the other queens, I took all of these photos before any of the queens were able to eat the food given to them, so I assume most of the inflated gasters are in preparation for laying eggs.









Colony B:  This queen has not laid eggs yet and is one of the queens I deemed to be a microgyne as she is the smallest or one of the smallest queens out of the nine. I would not be surprised if this queen was infertile, but we shall see. What's funny is despite her small size and possible infertility she was one of the most aggressive queens when trying to find pairs. She also continues to be aggressive/hyper-active during feedings and runs right for the food after I put it in. This is why the photos of the queen are her right next to the fruit fly, which helps to add scale for just how small she is (I'd put her at around 3mm). I have heard that some queens that do not mate just take on a worker role for their original colony. Given this queen's small size and aggression, this might be the case for this queen.






Colony C: This queen has also not laid eggs yet and I have yet to determine if she is a microgyne or macrogyne. In terms of size she is in the middle of the group and is close to the same size as the queens in Colony D. She is on the skittish side and displayed aggression toward other queens when trying to pair. As you can see from the second photo, her gaster is slightly inflated, so I'm hoping that's a sign that she will lay eggs soon.







Colony D: This is one of the two queen colonies. They have not laid eggs yet either. I would put both of these queens in the middle of the group in terms of size, with one being slightly smaller than the other. In terms of behaviour both of the queens are fairly relaxed and don't respond too much to light or vibration and readily accept food when I feed them. As you can see from the last photo, one of the queens (I believe the larger one) has a slightly inflated gaster, so hopefully they lay eggs soon too.







Colony E: This queen has also not laid eggs yet and is also one of the queens I deemed to be a microgyne. While she is not as small as the queen in Colony B, she is still quite smaller than the queen in Colony A and some of the other queens. Unlike the queen from Colony B though, this queen is extremely timid and is not a fan of any vibration or light. The moment I shine light on the test tube for feeding she pretty much freezes or tries her best to hide in the cotton as you can see from the photos. Given her timidness I deemed this queen to be one of the anxious ones.




Colony F: This queen has laid three eggs and is one of the queens I deemed to be a macrogyne. She is on the larger side, not as big as the queen from Colony A but pretty close. She also displayed hyper aggression towards other queens when trying to pair. During the feeding she seemed more curious than aggressive, but either way she was not very phased from the vibration or light. Apologies for the poor photos, I tried to get a picture of her holding an egg but she would not sit still. I look forward to the progress of this queen.








Colony G: This is the other two queen colony, which have not laid eggs yet. One of the queens is on the larger side and has a fairly inflated gaster, the other is on the smaller side. Given this I would peg the larger queen as a macrogyne and the smaller queen as a microgyne. This pair is more skittish than the queens from Colony D. They still readily accept food during feedings but take a longer time to get used to the vibration and light. These pictures are slightly better, with the fourth and fifth pictures making the larger queens inflated gaster clear. I expect this colony to have eggs soon, hopefully by the next feeding.









Edited by Karma, February 5 2024 - 5:37 PM.

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#3 Offline Karma - Posted August 4 2023 - 12:46 PM


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Update August 4, 2023:


In the two weeks since the last update, another queen has laid eggs. That means three out of the seven eight colonies now have brood. After closer observation, I determined that both queens from Colony G were not as cooperative as I previously thought and that their combined presence was causing more stress than it was worth. I decided to split these queens up, one of the queens will be Colony H from now on. The other five colonies have yet to lay eggs, which may mean they are either infertile or not comfortable enough in the current setup. I will continue to feed them until they die off just to be sure though.




Colony A: This queen continues to do well. She has laid another egg which is hard to see but if you look closely in the first photo by her back left leg you can see the egg poking out under one of the larvae. Additionally one of the larvae have pupated so she should be getting her first nanitic within the next two weeks.







Colony B: This queen has still not laid eggs but remains hyper-active and aggressive during any disturbance. I think there is a high probability that this queen is infertile but will continue to feed her. She could still be of use when I attempt to combine the colonies.






Colony C: This queen also has yet to lay eggs. She is still on the skittish side but readily accepts food.





Colony D: These queens also have not laid eggs. They are still fairly relaxed and readily accept food however they've put a lot of effort into digging into the cotton which could mean they are uncomfortable in the test tube setup. I have not decided if I am going to try a different founding setup with them yet, but if they do not lay eggs in the next couple weeks I will add a small piece of wood to the test tube setup and see if that helps.








Colony E: This queen has laid five eggs. Being one of the smaller and more timid queens, I deemed this queen to be a microgyne and didn't have many expectations but am glad to be proven wrong. I look forward to her getting her first workers.






Colony F: This queen has been an absolute beast in terms of laying eggs. Since the last update she has laid roughly seven more eggs which puts her at around ten eggs in total. Assumed to be a macrogyne with her larger size and increased aggression, I did have high expectations for this queen and I am glad she is living up to them. I look forward to seeing how many of these eggs she will actually be able to take care of and grow through the larval stage.







Colony G & H: As mentioned at the beginning of the update post, these queens have been separated. I did not take pictures after I separated them, but at the time they had still not laid eggs but were readily accepting food. I hope each of them will be more comfortable alone and lay eggs. 






Edited by Karma, August 4 2023 - 12:52 PM.

#4 Offline Karma - Posted November 12 2023 - 11:24 PM


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Update August 19, 2023:


I forgot to make this post when I first took the picture on August 19, but better late than never. Not a ton of changes since the last update, the queens that have been doing well are continuing to do well and the rest I am still waiting to lay eggs. As you'll see in the pictures, one of the queens got their first nanitic! With another queen not far behind. We are getting close to the end of egg laying season for these ants, so I assume if the queens that have not laid eggs continue to not lay over the next couple weeks, they will not lay at all until after hibernation. If they are fertile that is.




Colony A: This queen continues to be the best out of the bunch, and just got her first nanitic! Her brood continues to do well, with several of the eggs hatching into larvae. The large larvae have yet to become pupae. They may have paused their growth for hibernation and will not pupate until next year. I’ll watch them closely over the next several weeks.











Colony B: Not much has changed since last update. This queen has still not laid eggs.





Colony C: Ditto with this queen, who also has yet to lay eggs.





Colony D: These queens finally laid their first egg. I am not sure if they are going to lay more or if me exposing to light will stress them out too much, but just glad to see they laid a single egg. I am not sure if they will actually keep this egg given how close we are to hibernation season but we shall see.







Colony E: This queen has laid another egg since the last update, which brings her up to six brood. As you can see from the photos, her brood is at varying stages of development. With one large larvae, two medium larvae, two small larvae, and one egg. I look forward to her first nanitics if she is able to get them in time before hibernation.





Colony F: This queen has been doing well, growing the eggs she laid in the last two updates. As you can see from the photos, she has one pupae and several large and medium larvae. I am excited for her to get her first nanitics.






Colony G & H: I am grouping these two again as not much has changed since I split them. I had hoped that they would lay eggs but this still has not happened. Hopefully I will have more luck with them after hibernation.




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#5 Offline Karma - Posted November 12 2023 - 11:29 PM


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Update November 12, 2023:


I am making this update post late as well. I will date it for today but will include updates over the past two months in it. Some good news and some bad…


Things had gone great prior to me starting their hibernation. Most of the different colonies brood had stopped growing, except for Colony A which continued to lay eggs, and Colony F got their first two nanitics.


I put each of the colonies' test tubes in tubing insulation and then wrapped them in a blanket before putting them in a mini fridge. Given these ants ability to survive extreme cold temperatures, I wasn’t too concerned with what the fridge was set to. I normally set the fridge to as high a temperature as it will go, but I just left it on medium which I presume is somewhere around 5 degrees Celsius or 41 degrees Fahrenheit. I checked up on them a week later and all colonies seemed to be doing well except for two.


The queen from Colony B appears to be dead. Whether it’s because she was infertile, or because she drowned in some condensation that may have formed, I am not sure.


The queen and nanitic from Colony A appear dead as well. I am not quite sure why. I did mention earlier that this queen continued to lay eggs, so perhaps they were not ready for the cold temperatures I suddenly subjected them to and they succumbed to some sort of shock. Hard to say. I assume they did not drown like the queen from Colony B as they have more space in their test tube and they were dead in the middle of it near their eggs, with no water or condensation around. I took them out of the cold and left them for a day to see if they would move after heating up again, but they did not. I will leave them in their test tube and put them back into hibernation and pull them out with everyone else. I have seen some queens/workers stay completely dormant for up to a week after hibernation so I will give them a week after that to be certain that they are dead. I am highly confident that they are dead though, and have since transferred their brood to Colony D, with the two queens, who have readily accepted the brood and started caring for it.


I probably won’t update again until I pull them out of hibernation sometime at the beginning of March. I put them in hibernation around the middle of October, so that will give them around 4 months of hibernation.

Edited by Karma, February 2 2024 - 10:17 PM.

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#6 Offline Karma - Posted February 2 2024 - 10:17 PM


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Update February 2, 2024:


A week ago one of the queens in the two queen colony (Colony D) managed to get stuck in some honey so I had to pull them out of the fridge and perfom a rescue. The queen was unresponsive when I first noticed it and I was pretty sure she was dead. However, after pulling her out, washing her in water, then putting her back in the test tube and leaving her for a couple days, she started moving again. I had kept them out of the fridge during this so they were exposed to room temperatures. Apparently this was enough time for them to declare hibernation is over as the late stage brood given from the dead Colony A started to pupate. I then gave them a fruit fly which they happily accepted as well.


Since the brood started to grow and they were accepting protein, I thought that I would remove the rest of the colonies from hibernation. This would give them roughly 3-3.5 months of hibernation. I would ideally like to give more, but since this was only their founding year and they didn't lay many eggs, 3-ish months rest is probably fine for them. With that said, I do unfortunately have more deaths to report.


The queens from Colony C and Colony H did not make it through the "winter". Neither had any brood so it is still possible they were infertile.


With these deaths, only four of the original eight colonies remain. Colonies still alive: Colony D, Colony E, Colony F, and Colony G.


With the brood transfer to Colony D, all colonies have brood except for Colony G. Only Colony F has workers, but the Colony D and Colony E are not far behind with late stage brood.


No pictures in this update. I think I'll give them a week to acclimate to the warmer temperature and then start feeding them on a regular basis again.


Also, a colony I had sold last year was given back to me as the person is moving. Unfortunately they seemed to have been pretty heavily neglected. They still have a decent amount of workers so I think they'll make it. I may include them in this journal as an occasional "bonus species update" as I am bringing them out of hibernation at the same time.


The colony is one of the Camponotus herculeanus colonies found in my discontinued journal:




Will update some time in the next few weeks with pictures of all the colonies and their brood.

Edited by Karma, February 2 2024 - 10:26 PM.

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#7 Offline BleepingBleepers - Posted February 4 2024 - 8:44 PM


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Finally came upon this journal and wow, you put a lot of thought and effort into it. Lots of pictures and very detailed comments on what's going on with your Queens / Colonies.


Very impressive, I'll keep an eye on this, thank you very much for sharing. I'm quite overwhelmed really, it'll take me awhile to digest everything here, but great job!


As you know, I'm a bit wordy with my journal and love pictures and videos too so I can appreciate what you've done here as well. GL on your journal, Karma.

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JOURNAL: Camponotus CA02 - First Time At Ant Keeping CLICK HERE

JOURNAL: Ectomomyrmex cf. astutus - Ant Species #2 CLICK HERE

#8 Offline Karma - Posted April 13 2024 - 1:49 PM


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Update April 13, 2024:


Apologies for the lateness of the update, unfortunately the Camponotus herculeanus I received have been really struggling so I've been trying my best to disturb the ants as little as possible to get them going again. Many workers have died off in the herculeanus colony which brought them down to just four workers. Only bright side was they had eight+ pupae and several larvae and eggs, however four of the pupae were either eaten, dumped in the trash pile, or failed to pupate properly and died. Luckily two have successfully hatched recently and brought them up to six workers. The larvae and eggs continue to grow so I still have hope for them. I've ordered a Wakooshi S5 starter module for them and will be moving them into it in the next week or so. I normally would not move a colony into a larger nest so quickly but I have a feeling the test tube they are in just isn't working for them.


As for the Leptothorax colonies, all of the colonies have workers, however two of the queens I believe are infertile, more details about that below. The two queens who have been successful since the beginning continue to do well and grow their numbers.


This experiement is turning out to be much more difficult than I first thought. I originally started with nine queens, and now only two technically remain. I think it might be better to pivot from "combining colonies" to getting colonies E and F to the point of producing alates, then attempting to simulate a nuptial flight between the two and create a large or several large colonies from that. Of course to do this I need to get both of these colonies to the point of producing alates which will likely take at least another year. Anyways, on to the actual colonies ->




Colony E: This colony is doing great, I think with both this colony and Colony F their first two to three nanitics died off after about two months. This colony has three workers, two large larvae moments from pupating, one medium larva, and about six to ten eggs. They have quite an appetite and continue to accept whatever food I give them. Hoping to get them up to ten to twenty workers this year, then thirty to fifty next year and hopefully some alates.







Colony F: Similar to Colony E, this colony is doing great. Most or all of the nanitics have died off which has left them with four workers. Similar levels of brood as well, two medium larvae, and ten to fifteen eggs. Behaviour is similar to Colony E as well, although I'd say they are less restless/aggressive when giving them food. Similar timeline for this colony as well, hoping for alates next year. (Sorry for bad pictures).






Colonies D & G: I am grouping these together because they are acting the same way. As mentioned in the prior update, both colonies accepted the brood boost and eventually got workers, however they have been very restless and have refused to lay eggs. This makes me think both of these queens are infertile. I have continued to feed them and will keep monitoring. If they happen to stay alive until Colony E or F get alates, I will try to see if they will mate.

Edited by Karma, April 13 2024 - 1:56 PM.

Also tagged with one or more of these keywords: lepthorax, small ant, tiny ant, polydomy, polygyny

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