4/14/2017: Week 8 – Eight Horses and a Birth

Hey everyone, sorry this post is so late! I meant to post this last week but was unable to because I was traveling. 

Exciting news: I FINALLY GOT TO SEE ONE OF THE HORSES GIVING BIRTH!!!!!! It was an amazing experience, and I have a whole section of this post (in the middle) dedicated to it! Be sure to read all the way through to find out how it was! Anyway, I have a lot of things to share about Week 8, so let's get started!

Warning: Some of the pictures in the second section of the post (regarding the birth) are slightly graphic


LIFE ON THE RANCH

Remember the tree I talked about last time? Well, we got through a lot more of it this time around. I was also a lot more courageous than last time and helped remove some pretty gnarly branches. Some of them were pretty heavy too! We left the project nearly finished, and now have a huge pile of thorny branches next to the trimmed tree. After taking care of the tree, we also performed some bush maintenance near the border of our ranch.

Peep Mr. Ames Trimming 

Although we've left the huge pile of thorny branches for later, we took some satisfaction in knowing we were 100% done with the cleaning up near the border.

However, making progress on the tree wasn't even the most exciting part of that day. The best part was finally getting to see the mini horses, weeks after we'd first heard they would be coming in. As expected, they were super cute! In fact, because of their size, Mr. Ames lets them roam around in the yard like you would a dog, and it makes for some interesting situations.

Hey Little Fellas

In addition to these two joining the ranch, we had another surprise waiting for us when we arrived on Saturday. Including the minis, there were eight horses on the ranch! Eight! The previous week, there were only two, but now there were eight! All the stables were full and it was quite the sight to behold. Basically, two new horses had shown up on the ranch, and the two old horses had given birth. These two births were very unexpected and the interns didn't even hear about them until that Saturday. Apparently, one of the births led to a successful collection, but one was caught too late to collect any material. Anyway, there were now four regular sized adult horses, two mini horses, and two baby horses, for a grand total of eight. This was also my first time seeing a baby horse (because I've missed all the births) so I was pretty excited.

One of the Mother-Baby Pairs

The mother and baby pairs were returned to their owners later that day. We concluded Week 8 ranch duty by washing the two minis, one of which was quite stubborn. It was an interesting and productive day!


SUNDAY SURPRISE!

Now, what you (mostly me) have all been waiting for the past two months.

It all started while I was at a family get-together and I received a text from Mr. Ames. One of the horses (Mystic) had been put on red alert and he was asking if the interns could come for night watch. Of course, I showed up. It all started out quite normally. I got there around 12:45 (a little later than usual because I was busy). At about 1:00, I fired up the TV, opened Netflix, and laid back to watch an episode of 13 Reasons Why. But as soon as the intro sequence of the show ended, Nikash and I noticed the horse acting strangely, and suddenly, in a beautiful moment of relief and realization, knew we were about to witness this horse giving birth. After running to wake up Mr. Ames and going outside, we got to watch and help out with the process. 

How it All Began

In the picture above, you can see the hoof and the amniotic sac starting to come out of the mare. Around this time, we observed a lot of liquid coming out of the mare as its water continued breaking. Mr. Ames soon got in on the action.

There Comes the Foal!

Strangely, Mystic never sat down during the process. However, everything still went well, including the collection, although some amniotic fluid was spilled. As the baby fully emerged into the world, there was a lot of blood.

This Picture Has the Least Blood Showing

After Mr. Ames made sure the baby was healthy, he tended to the last part of the process: the passing of the placenta. He aided the process by tying a heavy cloth to the part of the placenta that had already come out. After the whole thing came through, Nikash and I checked it to make sure no pieces were left behind in the mare, for this can lead to some serious conditions. Complete horse placentas usually look like a capital F.

F for Placenta

Everything looked good! Afterward, Mr. Ames helped the baby horse stand up and learn to drink milk, while Nikash and I went inside and started washing the amnion. 

Yay!

Overall, it was a great experience, and well worth the wait! I'm happy I got to see it at least once before the internship ended, but I'm still looking forward to possibly being there for another. The first night watch where I ended up sleeping in my own bed was officially over! 

BACK TO THE LAB

Work in the lab started off very unexpectedly. Our first task on Monday morning was to get a huge package of styrofoam boxes out of a semi-truck and take it inside our lab using a pallet jack. Afterward, we used some of these new styrofoam boxes to prepare some ready to be mailed boxes. The remainder of the day was spent getting more liquid nitrogen and using it to figure out how to work the cell freezer in our lab. After tinkering with it for a while, I wrote a set of instructions on how to use it for a specific procedure meant for freezing equine semen. Here's a picture of me getting the liquid nitrogen:

Pretty Cool, Literally

On Tuesday, we liquified some more amnion in preparation for making our next products, AniMotion (which includes cells) and AniMatrix. We also checked the plates I mentioned last week, and found out everything was sterile! This means we can sell all of the products we've made so far, and continue working with the unfinished material we have. 

This was all usual, but we also did something new: learn how to clean a pipette! This is a valuable skill to know if you don't want to pay exorbitant amounts of money every time you want to get your pipette cleaned. Dr. Barcelo brought a set of pipettes from home for us to learn on, and we made sure to be extra careful cleaning around the delicate mechanisms and springs. Cleaning is important because it keeps the pipette calibrated and prevents cross-contamination of different solutions. We checked the calibration (using an analytical scale and water) of the pipettes pre-cleaning and post-cleaning, and all of them (except one, actually) were performing great. 

Don't Worry, It's Not Broken

Wednesday involved some new procedures, as we further prepared to make some AniMotion. Because this product includes cells, we had to count and make sure we were putting approximately 1,000,000 cells in each vial of product we made. We counted how many (both mesenchymal and epithelial) cells were in a small amount of the cell solution using a hemocytometer, and used math to figure out how many cells we had per unit of volume. The hemocytometer is essentially a microscopic checkerboard which lets you easily count cells using a microscope. 

Hemocytometer on the Microscope Stage

The blue tint you can see on the hemocytometer is due to a dye we used on the sample of cells. The dye lets us differentiate between living and dead cells. Once we had the counting figured out, we did some more math to establish how much of

1) Cell solution

2) Amniotic fluid

3) Liquified amnion

4) DMSO (a chemical which helps the speedy recovery of wounds) (very minute quantities go into our AniMotion product)

would be in each of the final vials of product. Once we knew all this, as well as how many vials of product we would be creating, the next step was creating a "cocktail" of the four ingredients listed above. However, we left out the DMSO, for the products must be frozen as soon as it's added, and we wanted to wait before actually packing the vials of AniMotion. But everything was now ready for when we would actually pack it. We did, however, pack a whole box of AniMatrix (which doesn't have cells or DMSO). 

And that sums up exciting, exciting Week 8! In a later post, I'll try to summarize all the products we've made so far and also talk about the sterilization issue I mentioned in the last post. But for now, thanks for reading, and see you next time!