Half Shell Hatch-A-Long! (Caution, unsuccessful, please read whole thread before attempting)

[Egg Snatcher]

I'm so sorry @MGG

 

[MGG]

I'm so sorry @MGG

Thank you.

 

[MGG]

Ok, what should I change the title to? I think that was a good idea.

 

[SnapdragonQ]

Ok, what should I change the title to? I think that was a good idea.

Nothing too drastic...Maybe just add (Caution) at the end, or (Unsuccessful)...or something to alert anyone who might be thinking of trying it. Hopefully they will read through the entire thread and it will dissuade them to try until/unless they have a specific plan?

 

[MGG]

Nothing too drastic...Maybe just add (Caution) at the end, or (Unsuccessful)...or something to alert anyone who might be thinking of trying it. Hopefully they will read through the entire thread and it will dissuade them to try until/unless they have a specific plan?

Ok, thank you!

 

[JaeG]

I've been doing a bit of research to try and figure out what went on and first I found this:

"In the embryo cultures where the embryos were transferred to the culture vessel after 55-56 h incubation, more than 90% of embryos survived until day 17 when a polymethylpentene film was used as a culture vessel with calcium lactate and distilled water supplementations. The aeration of pure oxygen to the surviving embryos from day 17 yielded a hatchability of 57.1% (8 out of 14)."

That's quite a low percentage of surviving chicks and that's with them being provided with pure oxygen.

I found this experiment as well which explains what goes wrong in shell-less experiments:

EXPERIMENTAL RESULTS

In an effort to maximize embryonic survivability, embryos have been cultured in three separate gas composition regimens (Fig. 3). In Experiment 1, in which embryos were cultured under normal atmospheric conditions, 100% survived for 5 days of incubation, 48% survived for 13 days and 9% survived for 18 days.

In Experiment 2, in which 5% C02 in air was applied at 8 days of incubation, 100% of the embryos survived for 7 days, 82% survived for 13 days and 9% survived for 19 days.

In Experiment 3, in which 5% C02 in air was applied at 6.5 days of incubation, and 8% C02 in air was applied at 12 days, 100% of the embryos survived for 5 days, 86% survived for 13 days and 5% survived for 20 days.

Many of the embryos in the latter two experiments developed to Hamilton-Hamburger Stage 40-41 (14-15 days, Hamilton, 1952). Apparently two major blocks to further shell-less development exist: 1) the chorioallantois does not completely enclose the egg contents in culture even by the 15th day, which it does in ovo by the 11th to 12th day (Romanoff, 1960), preventing the absorption of albumen into the amnion; and 2) egg shell calcium, which forms 75% of the skeletal mineral in the newly-hatched chick (Romanoff, 1972), is unavailable for skeletal ossification.

So I looked a little closer at the chorioallantois side of things and found a technique where they study the chorioallantois: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4583126/

Go down to figure 8 and you'll see the two techniques used - one where the egg contents are transferred to a dish and one where they are left in the shell. With the ones they leave 'in ovo' (ie. still in the shell) initially they only make a small incision into the egg and why is explained below:

The process of in ovo cultivation is initiated also at day 3 post-fertilization. At this time, a hole of approximately 3 mm in diameter should be created in the eggshell with a sterile tweezers and covered with a laboratory wrapping film to prevent dehydration and possible infections (Figure 8d). The eggs are then returned to the incubator with a relative air humidity of 65% and a temperature of 37°C in a static position until use (Figure 8e). This initial small incision changes the pressure inside the egg and prevents binding of the CAM with the shell membrane. At day 7 or later, the hole is extended to a diameter of approximately 3 cm in order to provide access to the chorioallantoic membrane (Figure 8f). This approach still enables experimentation through a window and it offers an almost unchanged physiological environment for the developing embryo.

I do feel that some kind of issue with the chorioallantois is the most likely explanation as to what went wrong because most got to around the same stage of development. And I'd read this as well: Even though the first lung respiration has started (after internal pip), the CAM remains important for respiration until up to day 20 of incubation.

I hope that helps some.

This has been a good distraction for me as it's day 18 of my incubation and it's driving me crazy! I have one that's been shadowing for over a day so things might start happening soon to put me out of my misery. It's amazing the difference between the eggs my DH picked up (a 3 hours drive away) and the ones that came through the post. All the eggs he picked up are still going strong despite 2 of them having cracks that I didn't find until at least halfway through incubation, but with the shipped ones I'm down to 3 out of the original 8.

 

[Clap p 1]

Hey MGG, what kind of incubator is your new one?

 

[MGG]

I've been doing a bit of research to try and figure out what went on and first I found this:

"In the embryo cultures where the embryos were transferred to the culture vessel after 55-56 h incubation, more than 90% of embryos survived until day 17 when a polymethylpentene film was used as a culture vessel with calcium lactate and distilled water supplementations. The aeration of pure oxygen to the surviving embryos from day 17 yielded a hatchability of 57.1% (8 out of 14)."

That's quite a low percentage of surviving chicks and that's with them being provided with pure oxygen.

I found this experiment as well which explains what goes wrong in shell-less experiments:

EXPERIMENTAL RESULTS

In an effort to maximize embryonic survivability, embryos have been cultured in three separate gas composition regimens (Fig. 3). In Experiment 1, in which embryos were cultured under normal atmospheric conditions, 100% survived for 5 days of incubation, 48% survived for 13 days and 9% survived for 18 days.

In Experiment 2, in which 5% C02 in air was applied at 8 days of incubation, 100% of the embryos survived for 7 days, 82% survived for 13 days and 9% survived for 19 days.

In Experiment 3, in which 5% C02 in air was applied at 6.5 days of incubation, and 8% C02 in air was applied at 12 days, 100% of the embryos survived for 5 days, 86% survived for 13 days and 5% survived for 20 days.

Many of the embryos in the latter two experiments developed to Hamilton-Hamburger Stage 40-41 (14-15 days, Hamilton, 1952). Apparently two major blocks to further shell-less development exist: 1) the chorioallantois does not completely enclose the egg contents in culture even by the 15th day, which it does in ovo by the 11th to 12th day (Romanoff, 1960), preventing the absorption of albumen into the amnion; and 2) egg shell calcium, which forms 75% of the skeletal mineral in the newly-hatched chick (Romanoff, 1972), is unavailable for skeletal ossification.

So I looked a little closer at the chorioallantois side of things and found a technique where they study the chorioallantois: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4583126/

Go down to figure 8 and you'll see the two techniques used - one where the egg contents are transferred to a dish and one where they are left in the shell. With the ones they leave 'in ovo' (ie. still in the shell) initially they only make a small incision into the egg and why is explained below:

The process of in ovo cultivation is initiated also at day 3 post-fertilization. At this time, a hole of approximately 3 mm in diameter should be created in the eggshell with a sterile tweezers and covered with a laboratory wrapping film to prevent dehydration and possible infections (Figure 8d). The eggs are then returned to the incubator with a relative air humidity of 65% and a temperature of 37°C in a static position until use (Figure 8e). This initial small incision changes the pressure inside the egg and prevents binding of the CAM with the shell membrane. At day 7 or later, the hole is extended to a diameter of approximately 3 cm in order to provide access to the chorioallantoic membrane (Figure 8f). This approach still enables experimentation through a window and it offers an almost unchanged physiological environment for the developing embryo.

I do feel that some kind of issue with the chorioallantois is the most likely explanation as to what went wrong because most got to around the same stage of development. And I'd read this as well: Even though the first lung respiration has started (after internal pip), the CAM remains important for respiration until up to day 20 of incubation.

I hope that helps some.

This has been a good distraction for me as it's day 18 of my incubation and it's driving me crazy! I have one that's been shadowing for over a day so things might start happening soon to put me out of my misery. It's amazing the difference between the eggs my DH picked up (a 3 hours drive away) and the ones that came through the post. All the eggs he picked up are still going strong despite 2 of them having cracks that I didn't find until at least halfway through incubation, but with the shipped ones I'm down to 3 out of the original 8.

Wow! Thank you so much for that! It was very interesting to read. Yeah, it probably was something with the chorioallantois. They all made it to day 19, and then quit before they got to day 20.
Lol! Good luck with your eggs! I hope they all hatch! That is quite a difference in the eggs. The PO must have been a little rough with them.

 

[MGG]

What kind of incubator is your new one

Oh, sorry! It's a Nurture Right 360.

 

[Clap p 1]

Oh, sorry! It's a Nurture Right 360.

Two Bresse have hatched. A hen then a roo. Yes you can tell. The roo 's legs are huge and his comb is pointy and above his head. She is fine boned and her comb is low to her head. The other two eggs have pipped so it looks like I'll have a good hatch.
seven is ready to eat. I feel like I have a new baby, but Seven sleeps all night.
And thank you, I thought that might be the incubator. How do you like it?