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Gulf Fritillaries Mating in Captivity

7/31/2016

We have finally succeeded in getting Gulf Fritillaries (Augraulis vanillae) to mate in captivity after failing multiple times to hand-pair them!

Over the past few weeks, we have tried on multiple occasions to hand-pair Gulf Fritillaries (Augraulis vanillae) using newly emerged butterflies, old butterflies and wild caught males and nothing would work. It didn't matter how well fed they were either. The males seemed particularly eager to mate with the females when they were together, but when forcibly put in contact with he female's genitalia, the male would not do the work no matter how stimulated they were. He is supposed to work his claspers and grab on to a place between the female's hooks and ovipositor that appear when her abdomen is squished (the end unfolds and becomes inverted). Either the male was confused and uninterested or the female was rejecting him.

After failing so many times to pair them, we eventually decided to try something different today. Since the males normally seemed so willing to mate normally, we thought that perhaps they would mate readily in captivity if provided with enough space. By today, we had already accumulated a total of nine sibling butterflies born from the eggs laid by a wild caught female. The sex ratios were a bit skewed; there were two males, one which was a few days old and the other fresh and seven fresh females. This afternoon we put all nine into our Mexican Bush Katydid (Scuderria mexicana) enclosure that measures 2 x 2 x 4 feet with walls made of window screening.

When we first put them in, we witnessed what could have been courting behavior. The males would hover around and encircle the motionless females, some of which would wave her abdomen upward (this is actually probably a rejection pose though, since she is revealing her abdominal glands by doing this which release repellents!). In nature, the male is supposed to patrol for females and hover around them before landing next to her and using his pheromones by flapping his wings in rapid bursts to waft the perfume to the female's antennae. We aren't exactly sure whether this happened, but when we checked on them a while later, the older male had paired! They were still together four a few hours.

Gulf Fritillaries lay fairly large eggs for their size and therefore the females probably have relatively few eggs in their abdomen. But we can probably hope to squeeze out a good 100 or so eggs if our newly mated female lays most of hers. Of course, we are still hoping for some more pairings and might even go out to collect a few more wild males if new males don't emerge to help out.

- Brian

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Hand-paired P. zelicaon Rearing Notes (7/18/16-7/31/16)

7/31/2016

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Rearing notes for the massive brood of Anise Swallowtail caterpillars (Papilio zelicaon) propagated from dozens of captive hand-paired butterflies from 7/18/16-7/31/16.

Rearing Notes 7/18/16-7/31/16:

7/31:

Transferred another 41 newly molted fifth instars onto the outside fennel enclosure; there should be 80 in total now
Ran out of fennel leaves to pick so fed flowers to a lot of the fourth instars

7/30:

Transferred 39 newly molted fifth instars onto two fennel plants outside enclosed by screening; plan to add more as the others molt into fifth instar
Took out 90 second instars to participate in the "Response to Phytochemical Stimulants for Feeding in Anise Swallowtails (Papilio zelicaon)" investigation
Almost all are fourth instars or third instars in apolysis; only a few third instars left

7/29:

Majority are now fourths or third instars in apolysis; some seconds and a few fifth instars, though many fourths are in apolysis
Will try to get the fifth instars to a sleeved plant outside so we won't be overwhelmed

7/28:

Took 30 newly molted third instar caterpillars out for the experimental group of the "Color Thermoregulation Mechanisms in Anise Swallowtail Caterpillars (Papilio zelicaon)" investigation
Recounted the entire stock again and it seems to be around 400

7/27:

Some fourth instars entered apolysis; some look a little undersized from the overcrowding
Tried thinning some of the dishes to avoid overcrowding; only 8-12 fourth instars per dish now
There is a single green morph fourth which is quite surprising since indoor reared fourth instar caterpillars have never been green morphs (too cool and humid)

7/26:

Many fourth instars now (maybe 60-70), a ton of thirds and some seconds and firsts

7/25:

More hatched today (50+) but that seems to be the end of it for now; the rest of the unhatched eggs are clearly infertile
Oldest dish now has newly molted fourth instars
On another note, one of our wild caught caterpillars turned out to be parasitized; a maggot emerged and spun a cocoon (last row middle)

7/24:

A bunch more hatched but didn't get a count (maybe 50-100)
Oldest dish caterpillars entering apolysis for fourth

7/23:

Around 30 more hatched today
Regrouped the caterpillars since many had molted to second instar or third instar; there are not 13 dishes; the first four dishes have 24 third instars each; the fifth and sixth have 36 second instars in apolysis each; the seventh and eighth have 36 second instars each; the remaining five have around 48 first instars each

7/22:

Just a single hatchling today; there was a gap in oviposition by the female butterflies because most all of them were depleted of eggs at one point before we cycled in new ones
A few more dead in each dish again as always though not very many today; maybe 10-20 in total which is not bad for 12 dishes
Didn't bother regrouping caterpillars today (alright fine, we were too lazy)
The caterpillars in the first six dishes are now all in second instar, with the first dish having mostly apolysing ones; the other six dishes still have first instars

7/21:

Another 40 or so caterpillars hatched- that looks like almost the end of it for at least a few days
There were a few more dead caterpillars from each dish again which collectively added up to 40-50; this time it probably wasn't so much the rough handling when tranferring food but desiccation since a lot were in a dish that had a very dry, wilted leaf
Regrouped remaining caterpillars into 12 dishes; first three have roughly 30-40 second instars, remaining have 48 first instars except last one only has 38 but is curreclty being added to as more caterpillars hatch

7/20:

Another 110-120 more caterpillars hatched
Regrouped all the caterpillars along with the newly hatched ones so that each dish contained the individuals that were closest in age which is always important when rearing several caterpillars per container (otherwise smaller individuals will lag increasingly more behind and the larger ones will get increasing more ahead)
There were a few dead caterpillars per dish that added up to probably at least 30 dead in total; most were probably a result of handling too roughly when transferring to a new leaf
There are now a total of 14 dishes now; the first two have 36 second instars each, the third has 36 first instars in apolysis, the remaining have 48 first instars each except for the last one that only has 12 and is currently being added to until it reaches 48
Placed the dishes into a tank and put in under two 50 watt heat lamps at night just to warm things up a bit in order to make them grow faster; since there are so many, the faster they grow the better so we don't have to care for them as long (fennel's been harder to pick these days since most were cut down by city maintenance on 7/18)

7/19:

Not as many caterpillars hatched today- only around 60-70
We filled up the rest of the 14 caterpillar dish to around 48 caterpillars and started on a new dish that has around 30 and counting
Went out and collected eggs from the potted plant oviposition enclosure for the first time in several days; didn't get exact count but its definitely a few hundred

7/18:

A whopping 200+ more caterpillars hatched over the course of the day and counting
We regrouped all the caterpillars from this brood into 10 petri dishes; two contain ~24 of the oldest ctaerpillars each; seven contain the remaining most of the remaining 300-400 caterpillars with ~48 caterpillars each; the last dish contains 14 of the newest hatchlings so far and is being added to as more hatch

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More Unidentifieds: True Bugs

7/31/2016

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Today at Albany Hill, we discovered a colony of interesting looking true bugs (Hemiptera) of the Leptoglossus genus.

Identifying Hemiptera is a headache, especially for people who have never actually studied them like us. Here are some pictures of a colony of true bugs we found at Albany Hill on bramble (Rubus ursinus). They are definitely leaf-footed bugs (Coreidae) and are probably in the Leptoglossus genus, but the exact species we cannot determine.

- Brian

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Eggs on Coast Live Oak- Duskywing (Erynnis spp.) Eggs?

7/30/2016

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We found a couple of nice looking eggs on Coast Live Oak (Quercus agrifolia) today at Albany Hill, but are having a hard time identifying them! So far we suspect a duskywing species (Erynnis), but there are several in our region that specialize on oaks.

Albany Hill is, without a question, Albany California's greatest landmark for wildlife. After all, the rest of the city is suburban; and yet, we have only investigated the place a few times since it is far from where we live. The few times that we have been there, we were collecting Western Tiger Swallowtails (Papilio rutulus). On those trips, we encountered several other lepidopteran species (well, butterflies since we have always gone in the day) such as the Common Buckeye (Junonia coenia), Monarch (Danaus plexippus), Mourning Cloak (Nymphalis antiopa), Red Admiral (Vanessa atalanta) and other Vanessas, Cabbage White (Pieris rapae), Umber Skipper (Poanes melane), Fiery Skipper (Hylephila phyleus), among quite a few other smaller butterflies (mostly nymphalids and lycaenids).

Today, we decided to venture back to Albany Hill not with any of these particular species in mind; we would simply catch whatever we thought would be worth catching. Like many summer days in the East Bay, today was quite cool and cloudy, with the temperature in the mid 60's and humidity around 50% when we went out around 2 PM. This was not at all good weather to hunt for butterflies since most such diurnal insects are programmed to roost when day time weather is unfavorable like this, but there was a hint of sunlight earlier so we took our chances. When we arrived, we could not see a single flying insect in sight -- not one butterfly or even a damselfly which we saw dozens of on the other trips. It was a bad call.

Nonetheless, we stayed around just in case we should see something. We found a Monarch roosting in a tree that darted out when we came by because me must have startled it. We also caught a Fiery Skipper that we immediately released because we have no use of such a species. We actually spent the bulk of our stay investigating the vegetation for eggs or caterpillars. Since the part of Albany Hill we were at was bordering Cerrito Creek, it was an excellent riparian area dense with willow (Salix), a host of a ridiculous number of lepidopterans in our region including a few of those that we have encountered at the hill. Mixed in were also a lot of evergreen coast live oaks (Quercus agrifolia), a much less popular host due to its divergence from its deciduous relatives (the foliage of many evergreen species, such as coast live oak, have evolved to be heavily defended against damage). None of the species we have encountered utilize it.

As we were searching, we found a single very young oak tree growing on the creek bed. It was a small tree and was actually putting on a substantial amount of new growth for this time during the season probably because the creek provided the necessary water (none of the trees growing away from the creek had any new growth whatsoever). There were several new, dark pink shoot tips. But that was not what initially interested in this plant; when we had first stepped upon it, we noticed a few fungal growths that we originally mistook for eggs (alas!). These fungal growths were more or less spherical, had a slight mottled appearance, could be easily detached from the leaf, and closely resembled some kind of Saturniid ova. However, when we looked closer, we realized that they could not possibly be eggs because they came in all different sizes, presumably because the smaller ones grow into the larger ones. Moreover, breaking them open did not yield any distinct egg shell or yolk piece at all; it was the same material all the way through the entire structure and it felt slightly rubbery rather than containing fluid.

As we were investigating these fungal growths, however, we noticed a bright yellow dot, conspicuously located on one of the pink new growths which would become the highlight of this trip. This time, it really was an egg, presumably lepidopteran. It was about the size of aVanessa egg, very spherical, and ribbed (shown below). Upon closer examination, we collected four more eggs, laid exclusively on the tender leaves of the new shoot tips. When we were leaving Albany Hill, we noticed another, much larger oak tree on the other side of the creek that was also putting on new growth and discovered two other eggs. One was blackened on one side and may be dead while the other one is a dark orange and is likely in a later developmental stage than the other bright yellow ones.

As discussed above, the evergreen coast live oak is a terrible host plant for most insects and it was the last tree at Albany Hill that we expected to find any eggs on. To defend against herbivory, this particular variety has incredibly tough and dry leaves adorned with spikes and seems quite indigestible. No wonder whatever laid the eggs has a preference for the tender new growths!

Upon returning, we flipped through our Lepidoptera field guides and did some quick internet searches to identify the eggs. Statistically speaking, most Lepidoptera found in the wild are moths as they constitute the majority of the order, but in this case we suspect a butterfly because the eggs were laid singly and strictly on new outer growing shoots which just doesn't seem very moth like (moths tend to be much more careless about where they lay eggs). So, we first looked at true butterflies (Papilionoidea), and the only few species in our region that specialize on oak are the California Sister (Adelpha californica) and a few random hairstreaks (Lycaenidae). However, we have never seen any California Sisters on any of our trips to Albany Hill and their eggs barely resemble ours, and the hairstreaks are far too small to lay eggs the size of ours and their eggs don't resemble ours anyhow. So, next we moved on the skippers (Hesperioidea), and we quickly found two likely candidates in the Erynnis genus (Duskywings): the Propertius Duskywing (E. propertius) and the Mournful Duskywing (E. tristis). They are very similar species that both commonly utilize live oaks and lay their eggs on the newest shoots. A few quick internet searches showed us eggs that look very similar to ours, though it is difficult to say for sure because they're so small are hard to see. Anyways, for now our best bet is that the eggs are that one of these two Duskywing species, though if they really are we probably won't be able to tell which one it is until they are adults.

- Alan, Brian

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Response to Phytochemical Stimulants for Feeding in Anise Swallowtails (Papilio zelicaon) Investigation Notes

7/30/2016

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We are currently conducting an investigation to see if there is a difference in Anise Swallowtail caterpillar growth rate or overall size (in mass) when fed fennel (Foeniculum vulgare) cuttings coated in different phytochemical stimulants: fennel (Foeniculum vulgare) oil, basil (Ocimum basilicum) oil, and anethol. The control group is 30 caterpillars obtained at the second instar and fed non coated fennel cuttings. The experimental groups are three groups of 30 caterpillars reared under the same conditions except they will each be fed fennel cuttings coated in one of the three substances. These notes will aid us in writing an article on this experiment.

Investigation Notes:

7/30:

Took 120 second instar caterpillars from the hand-paired batch to participate in the control and experimental groups
Coated the three experimental group's food with one of the three substances

- Alan, Brian

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The Tilden Painted Lady Has Eclosed

7/29/2016

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The Painted Lady caterpillar (Vanessa cardui) that we collected from Tilden Regional Park (Berkeley, CA) on 7/9 has eclosed into the adult.

In exactly ten days since it pupated, the fifth instar caterpillar that we found on milk thistle (Silybum marianum) at Tilden Regional Park has eclosed into the adult (such speed that is!). It is a female.

The top row shows the very pharate chrysalis, just hours before it eclosed. The next two rows show it drying its wings in the sun while resting on a milk thistle in our yard. The last row shows a rare moment where it has opened its beautifully patterened wings; normally, the rest with their wings closed upright.

- Brian

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P. zelicaon Chrysalis Color and Size (Mass) Variation

7/29/2016

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Out of curiosity, we decided to compare some of our hundreds of Anise Swallowtail chrysalises (Papilio zelicaon).

From left to right: Western Tiger Swallowtail (Papilio rutulus); 1.5g brown morph Pipevine Swallowtail (Battus philenor), 1.5g brown morph Anise Swallowtail (Papilio zelicaon), 1.4g green morph Pipevin Swallowtail, 1.5g green morph Anise Swallowtail

The left image compares a 1.5g Anise Swallowtail chrysalis to a 0.8g, the respective minimum and maximum of our collection. The middle image shows 9 chrysalises with a 0.1g graduation from 0.8g to 1.5g. The left image shows two 1.5g chrysalises next to a penny.

These two photos show, in different lighting, the color range of Anise Swallowtail chrysalises from dark brown (has black stripes) to tan to bright green.

- Brian

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Color Thermoregulation Mechanisms in Anise Swallowtail Caterpillars (Papilio zelicaon) Investigation Notes

7/29/2016

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We are currently conducting an investigation to see if we can induce green morph fourth instar Anise Swallowtail caterpillars (Papilio zelicaon). The control group is 30 caterpillars obtained at the third instar and reared indoors on fennel (Foeniculum vulgare) cuttings. The experimental group is another 30 caterpillars reared under the same conditions except they will be placed outdoors. These notes will aid us in writing an article on this experiment.

Investigation Notes:

7/29 (Day 1):

Experimental:

Put the dish outside; the caterpillar became much more active due to the heat

Control:

Kept inside as normal

- Alan, Brian

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S. ophthalmica L4 Rearing Notes (7/22/16-7/28/16)

7/28/2016

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Rearing notes for our fourth instar Ophthalm Sphinx caterpillar (Smerinthus ophthalmica) from 7/22/16-7/28/16. It was originally found as an egg mistakenly laid by the moth on Shasta Daisy (Leucanthemum x superbum) growing under a willow tree (Salix), the intended host, at the Eastside Permaculture Garden (Albany, CA) and has been reared indoors on willow cuttings.

Rearing Notes 7/22/16-7/28/16:

7/28:

Seems to be reaching its peak of this instar; it ate a lot overnight but has only gained .4g and still looks to be around the same size, maybe a little plumper
By nighttime, the caterpillar seemed to have stopped eating
Stats:
- Mass: 4.6g (2:00 PM)
- Length: ~2.5 inches neutral position (4:00 PM) and ~3.0 inches elongated

7/27:

We're really getting confused now due to the enormous size of this presumable fourth instar Ophthalm Sphinx caterpillar; it is already 2.5 inches long relaxed and 3 inches stretched and is almost half an inch thick which are approaching the measurements for a full grown fifth instar
We are not sure if this species only has four instars (seems rather unusual) and most sources mistakenly assumed it had five, or if our caterpillars is indeed a fifth instar and we somehow missed it when it molted at some point (yet this seems quite impossible since we have taken photos daily, kept the head capsules of every instar, and have observed it very closely)
Since we aren't certain if this is the final instar (what ever instar it is), we put the caterpillar with the willow cuttings into a plastic tub in case it begins wandering to find a pupation site
Stats:
- Mass: 4.2g (2:00 PM)
- Length: ~2.5 inches neutral position (4:00 PM) and ~3.0 inches elongated

7/26:

This caterpillar is getting so large and eating so voraciously that it's hard to imagine it still has another instar after this; in fact, S. ophthalmica (or cerisyi) are not extremely large Sphinx moths (forewing length 34-47mm [1.34-1.85 in]) and most sources say the full grown caterpillar is 3 inches, but our fourth instar already measures around 2.25 inches in a relaxed, non-elongated position; not sure if this is normal or if ours is unusually large
Stats:
- Mass: 3.6g (4:00 PM)
- Length: ~2.25 inches neutral position (11:00 PM)

7/25:

Needless to say, its grown incredibly larger than yesterday and feels very smooth and firm
It's anal horn looks even bluer than before; fifth instar cuticle on it way developing perhaps
Went on an eating rampage as usual, stripping the two cuttings given last night pretty badly; 13 and a half leaves eaten out of the 20 leaves in total
Stats:
- Mass: 3.0g (6:30 PM)
- Length: ~2.125 inches neutral position (2:00 PM)

7/24:

This caterpillar's appetite has gotten ridiculous-this morning at around 3:30 AM (yes, AM), we heard it crawling around since it is in our bedroom now and saw that it was doing so because it had finished nearly the entire cutting given to it during the day so we got out of bed and got it a new cutting
When we woke up, it had completely finished the cutting given to it yesterday that had five leaves and a few new shoots and had already started on the one given to it at 3:30 AM
By nighttime, it had stripped four of six leaves plus the few new shoots on its cutting, so we fetched it two large cuttings that should last the night
Its anal horn is becoming more blue on the dorsal side but is still pinkish purple on the ventral side
Stats:
- Mass: 2.1g (4:00 PM)

7/23:

Okay, how many times to we have to say this when it comes to this Ophthalm Sphinx caterpillars?; it's grown incredibly since yesterday and has a voracious diet, eating a good chunk of a leaf almost every few hours in just the fourth instar
This is no joke, but we weighed the caterpillar again at around 3:00, and it shot all the way up to 1.3g from the .5g that it was yesterday at 4:00 PM, almost triple the size in less than a day; seems it hardly put on any weight during the first three instars and the true growing is starting now
Got a new cutting and it immediately began eating away at a leaf even though it was resting before; perhaps the freshness of the leaf stimulated its appetite
Stats:
- Mass: 1.3g (3:00 PM)

7/22:

Already looking much plumper than when it first molted after just one day in fourth instar
Interestingly, although we thought that it would grow out of the preference for new, tender leaves now that it is so large and is able to take on the mature leaves with ease, it still crawled up to the top of the cutting and ate all the new growth before starting on the more mature ones; perhaps it is not a preference for the new growth but rather it has a habit of eating from the top of the branch to the bottom which is definitely possible since the Cecropias do it sometimes
Took it out of the cabinet at around noon which is way too late because the sun was probably up at around 6:00 AM, but extra darkness probably won't hurt; we put it into a dark room at 9:00 PM tonight instead of the cabinet so that the room will light up by itself at the natural time so that we don't have to remember to take it in or out of the cabinet at the right times
Got a new cutting; going to start changing cuttings daily now since it eats more
Stats:
- Mass: .5g (4:00 PM)

- Alan

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Polyphemus Caterpillar Ecdyses into Third Instar

7/28/2016

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Our oldest second instar Polyphemus caterpillar (Antheraea polyphemus) has ecdysed into third instar.

At around 3:00 PM, we checked on our Polyphemus caterpillars and saw that the one that had entered apolysis on 7/26 looked pharate (two cuticles in one), with the third instar hairs clearly visible as black streaks on the thorax as seen below.

After waiting for about half an hour, the caterpillar inhaled air and began ecdysis. We started filming the process, but the camera's memory card maxed out before we could finish (should've cleared it out first!). A little more than half the caterpillar's body is out of the old cuticle by the end of the video, but we made sure to get shots of it when it was completely done as well.

Second Instar Polyphemus Moth (Antheraea polyphemus) Caterpillar Molting*

* Shown in 4x speed

The third instar caterpillar looks nearly identical to the second instar caterpillar which looked similar to the first instar as well. The only subtle changes are that the oblique lateral lines that connect the two rows of lateral tubercles is a little more obvious now and is yellow in color.

Later in the day when we changed the food, we decided to transfer the third instar onto a leaf stuck into a water bottle rather than put it back in the petri dishes with the others and plan to do this with the others after they molt into third instar.

Second instar wasn't not quite as long as first instar, taking about 6 days and 1 hour from 7/22-7/28. First instar took roughly 7 days and 2 hours from 7/15-7/22. Below is the count for today.

L2+ max mass: 0.15g
Count: L3: 1; L2+: 19; L2: 13; dead: 5 (1 died shortly after birth, 4 unknown)

- Alan

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