The project I'm working on has been a long time coming; clouded leopards have very unique reproductive problems, unfortunately which cannot be fixed with the snap of a finger (were this the case, I'm pretty sure I'd be famous right now). I'm working on the female side of things, but there is also a male aggression study going on that a new intern just arrived for. I'm working for a pretty cool lady named JoGayle Howard. JoGayle is quite literally the clouded leopard queen: she's headed up most of the studies and the advances in clouded reproduction. She's a theriogenologist, or a veterinarian who focuses on reproduction, and a PhD at the Smithsonian. She's done everything, from developing successful cryopreservation of sperm, laparoscopic artificial insemination in carnivores, and genome banking. In short, she's pretty awesome. Which by default, the project I'm working under, under the supervision of postdoc Rebecca Hobbs, is also awesome. For more insight into the project, check out this website:
http://cloudedleopard.org/default.aspx?link=research_inzoos
Having been here in Front Royal for a month, I've finally gotten into the swing of things with the project. I've cataloged, dried, and sorted more fecal samples than I care to know, and developed a system for which to extract each sample set. I'm using a boiling extraction process, which is pretty simple: fecal sample are collected from each animal over a pre-determined time period (one sample, or one week's worth, is not enough to quantify hormone changes in reaction to the study protocol); freeze-dried of all water (called lyophilization); and crushed and weighed out into test tubes. Once in the tubes, I add radioactivity that will later determine the extraction efficiency of each sample; that is, the radioactivity provides a way to show how much hormone was pulled from the fecal sample during the extraction process. This is essential because later steps will be measuring hormone levels, such as estrogen and progestogen. The extraction of this whole process takes place with the addition of ethanol and boiling the mixture for twenty minutes (smells great), then centrifuging (think worst merry-go-round ride ever), then centrifuging some more with some more ethanol, and drying down the supernatant two different times. There are more in-between steps, with some methanol and dilution buffer and a sonicator, the machine that cleans jewelry, but these are the basics. After I have my final dilution, called a cocktail (no joke!), I throw all the samples into a beta counter, a machine which measures the radiation emitted by beta-emitting nucleotides via light pulses (the scintillation fluid I add to the cocktail throws a nice prism ). In short, I find out if I did the whole process right and have proper extraction efficiencies for my samples. Usually, I do. I've got to re-run about twenty samples, and so far I've run almost 600. Once I'm done extracting all these samples, I'll hopefully have time left during the internship to run enzyme immunoassay plates (EIA) for the hormones. I'm looking forward to more procedures under the guidance of the lab manager and the dugong project being run by a visiting scientist from Australia. Consequently, I see a lot of this:
I guess this makes me a lab rat!
I've also looked into volunteering with the clouded leopards on site. I've had a good peek at two eight-month old leopards that are visiting, and I'm not sure there is anything more beautiful, or adorable, than these rascals. Although the clouded animal keeper, Jessica, might think otherwise of the very-demanding squawking these two cats can emit. I can hear it all the way down the hall!
Finally, a few of us went hiking in the Shenandoah last weekend. We had so much fun! Martin is a GIS intern who just arrived from Germany, so we took him out to see the last of the changing leaves. Two hikes and 7.5 miles later, we had seen a gorgeous peak and a lovely waterfall. I wondered why I was so exhausted during the first hike (it's not like I hiked every day this past summer), but then I realized that we were on the hike that, in less than a mile, you climb 900 ft in elevation. That's a lot of climbing and hating, in case you were wondering how to measure that. Once we got to the vista, however, all huffing and puffing was lost in the view of the Shenandoah valley in the fall. Scaling huge rocks, we sat atop the valley for almost an hour, taking in the view and having a Lion King moment or two (think Pride Rock).
http://cloudedleopard.org/default.aspx?link=research_inzoos
Having been here in Front Royal for a month, I've finally gotten into the swing of things with the project. I've cataloged, dried, and sorted more fecal samples than I care to know, and developed a system for which to extract each sample set. I'm using a boiling extraction process, which is pretty simple: fecal sample are collected from each animal over a pre-determined time period (one sample, or one week's worth, is not enough to quantify hormone changes in reaction to the study protocol); freeze-dried of all water (called lyophilization); and crushed and weighed out into test tubes. Once in the tubes, I add radioactivity that will later determine the extraction efficiency of each sample; that is, the radioactivity provides a way to show how much hormone was pulled from the fecal sample during the extraction process. This is essential because later steps will be measuring hormone levels, such as estrogen and progestogen. The extraction of this whole process takes place with the addition of ethanol and boiling the mixture for twenty minutes (smells great), then centrifuging (think worst merry-go-round ride ever), then centrifuging some more with some more ethanol, and drying down the supernatant two different times. There are more in-between steps, with some methanol and dilution buffer and a sonicator, the machine that cleans jewelry, but these are the basics. After I have my final dilution, called a cocktail (no joke!), I throw all the samples into a beta counter, a machine which measures the radiation emitted by beta-emitting nucleotides via light pulses (the scintillation fluid I add to the cocktail throws a nice prism ). In short, I find out if I did the whole process right and have proper extraction efficiencies for my samples. Usually, I do. I've got to re-run about twenty samples, and so far I've run almost 600. Once I'm done extracting all these samples, I'll hopefully have time left during the internship to run enzyme immunoassay plates (EIA) for the hormones. I'm looking forward to more procedures under the guidance of the lab manager and the dugong project being run by a visiting scientist from Australia. Consequently, I see a lot of this:
I guess this makes me a lab rat!
I've also looked into volunteering with the clouded leopards on site. I've had a good peek at two eight-month old leopards that are visiting, and I'm not sure there is anything more beautiful, or adorable, than these rascals. Although the clouded animal keeper, Jessica, might think otherwise of the very-demanding squawking these two cats can emit. I can hear it all the way down the hall!
Finally, a few of us went hiking in the Shenandoah last weekend. We had so much fun! Martin is a GIS intern who just arrived from Germany, so we took him out to see the last of the changing leaves. Two hikes and 7.5 miles later, we had seen a gorgeous peak and a lovely waterfall. I wondered why I was so exhausted during the first hike (it's not like I hiked every day this past summer), but then I realized that we were on the hike that, in less than a mile, you climb 900 ft in elevation. That's a lot of climbing and hating, in case you were wondering how to measure that. Once we got to the vista, however, all huffing and puffing was lost in the view of the Shenandoah valley in the fall. Scaling huge rocks, we sat atop the valley for almost an hour, taking in the view and having a Lion King moment or two (think Pride Rock).
Hike two was longer, but easier, and took us along a beautiful creek down to a waterfall. Instead of hiking to the cliff overlooking the lengthy fall, we opted to hike down into the midst of it. The slope wasn't very steep, but the view and the sounds were beautiful:
All in all, so far so good. I must get to bed- I've got to get into the lab early tomorrow!