Sunday, July 09, 2006

 

Cholinesterase

I spent two evenings during the week and most of the day Saturday (yesterday) in the lab, but I finally got through all of the samples that I brought back with me (78 of them).

Kimberlee is going to try to send 25 more down this week (some animals that are being net-gunned and radio-collared this weekend), and that will be the last of them.

That would bring the grand total to 128 samples, which is 28 more than originally planned, but hey, the more the merrier! I just hope I'll have enough grant money left over to pay for them!

Here I am in the tox lab working on one of many samples. I start with frozen whole blood, and once it thaws it gets diluted with a pH 8.0 phosphate buffer (10 uL blood and 10 mL buffer). The lab has this nifty machine that does the dilution automatically, whoever invented it is a genius! Once it's diluted (see test tubes below) it sits for 5 - 10 minutes while the red blood cells lyse. We let them lyse so that the cholinesterase in the red cells is measured too.


Once the tubes have sat for a few mintues, the dilution gets put into cuvettes that get placed in the spectrophotometer. I do all the samples in duplicate, plus a standard and a control, so there are 4 cuvettes for each sample.

Here they are in the chamber of the spectrophotometer, just waiting to be read! The clear one is the control, its human plasma with a known amount of ChE. The chamber to the left is the standard, it doesn't get any reagent added, and the spectrophotometer measures the samples against it.

Once they're in there I add a couple of reagents, mix, and hit the start button. It takes 6 minutes to do each sample, and it prints the results immediately afterwards. I think it's neat. There are some smart people out there that have figured out how to do all of this stuff, and they make it user friendly so that people like me can do projects like this.

My diagnostics rotation is moving right along too. This week I was on cytology, so I sat with the clinical pathologist every afternoon while she interpreted impression smears, aspirates, bronchio-alveolar lavages, trans-tracheal washes, and other exciting things.

I got to see some blastomycosis, an aspiration pneumonia, and some sarcomas, carcinomas, and lymphomas. It was great learning for me, the pathologist would give us a few slides to look at before we started for our interpretation. I was right more than half of the time, but I still have a lot to learn! Especially about lymph node cytology.

This week I go back to necropsy, hopefully there will be some interesting things. I doubt there will be any caribou or grizzly bears though.

Sunday, July 02, 2006

 

Back in Illinois

Well here I am back in the flatlands of Illinois. I got back Sunday night and I started my diagnostics rotation at 9:00 on Monday morning. I thought that this rotation would be much less time consuming than my first rotation (small animal internal medicine), but that's not exactly the case. I am definately at school less (8 - 10 hours a day), but there are necropsy reports to write, "homework" (clin path cases), case correlations, and presentations to prepare for. Overall it's much less stressful than medicine though (mostly because the animals we see are already dead). My first week of diagnostics I necropsied a swan, a Jersey cow, a draft horse, and a mouse.

I had planned on starting my cholinesterase samples on Tuesday, but I kept ending up a school late writing reports. So I ended up in the lab all weekend chugging through my samples. I got 18 of them done yesterday (saturday), and I'm hoping to get through another 20 or so today. In all I have 87 samples to do, so it will take me a few weeks of evenings and weekends to get them done.

This picture is the machine that reads the cholinesterase activity of my samples. It's a Shimadzu 160-UV spectrophotometer (housed in the college of vet med's diagnostic toxicology lab). What it does is send a beam of UV light through my samples (after I have diluted them and added a couple of reagents), then it reads the difference in absorbance based on a standard. It takes about about 6 minutes to do each sample, plus it has to warm up for a while, the samples have to thaw, I have to add the reagents, etc.

I'm actually writing this now as I'm waiting for my samples and the reagents to thaw out. I will be here for a few hours still. Tomorrow it's back to diagnostics, and hopefully I'll have some time in the evenings to do a few samples.

Friday, June 23, 2006

 

Last Day in AK

It's not really my last day, I don't leave until tomorrow night, but it's my last day working at ADFG. I'm sad to be leaving, but I'm excited to start the assays on my blood and to really start digging into my project.

Here's just a picture (from yesterday) of the tiniest necropsy yet this summer - an 8 gram little brown bat. It was so small that we did it under a microscope. It can be a challenge to switch from working on something as big as a caribou to something as small as a bat, but that's one of the reasons I love veterinary medicine.

This morning it's smokey in Fairbanks, there have been a couple of wildfires burning not too far away. Every summer tens of thousands (or maybe even hundreds of thousands) of acres of forest burns in the state. That sounds alarming, but it's a necessary part of a healthy ecosystem, and unless it's threatening villages or people, the firefighters leave them alone. In fact, ADFG (and other agencies) often start fires on purpose to manage wildlife habitat.


I started the day like I have for the last few days by picking Ostertagia out of abomasums. I only did that for a couple of hours though, because two necropsy specimens, common ravens, came in. The first one was in great shape, not rotten at all. It had some pretty significant bruising on it's skull, but no other lesions consistent with trauma or disease.

Any raptors or corvids (crows, jays, and magpies) that die up here automatically get samples taken to test for West Nile Virus (WNV). That involves taking three small pieces of brain. After I took the brain samples I swabbed the birds cloaca for Avian Influenza testing. As you may or may not know, Alaska is the frontline for avian flu monitoring in the United States. Since Alaska is only 60 miles from Russia via the Bering Strait, and many birds migrate between the countries, it is feasible that avian flu will arrive to the U.S. through Alaska. I also took a spleen sample for the UAF tularemia project. Since it was in such good shape, I took some samples for histology too.

The second raven, unfortunately was extremely decomposed, so I wasn't able to take any samples. But he did have two severely broken legs, so I'm sure he was hit by a car or something.

After the ravens, I got back to work on the abomasums for a bit, and then another bird came in.

This one was very exciting for me. It was a great gray owl, the largest owl in North America. I had never seen one before (not even a dead one). It was found by a resident of Delta Junction (a town about 100 miles from Fairbanks), and brought to the local veterinarian. The vet took some radiographs, and it had a fractured clavicle. Unfortunately that injury would most likely prevent this owl from ever flying (even if it healed well), so the veterinarian euthanized it and sent it here.

It was a very fresh specimen, so I took the same sample as the raven, brain for WNV (see left), a cloacal swab for avian influenza, spleen for tularemia, and tissues for histology. I also just checked everything out to make sure there was no disease present. It looked good, except for some deep bruising in the pectoral muscles (probably caused by whatever broke the clavicle), and hemorrhage in the left lung (also probably from the same event).

So that's it! It was a good last day, I got to do 3 necropsies. Tomorrow I will pack up my stuff, head to Kimberlee's for a barbecue, and then get on the plane to head back to Chicago. I'll get to Chicago Sunday afternoon and make the 3 hour drive down to school. I'm not going to post again until Monday. I've had a fabulous time here in Alaska, and I can't wait to come back!

 

Fox

6/22/2006

On her way in to work this morning Kimberlee found a red fox kit on the side of the road. She was sure it was hit by a car, but she brought it in for me to necrposy, since it's a great learning experience, and we wanted a piece of spleen for a study at the University of Alaska (on Tularemia).

It was tiny (only 5 lbs) and very fresh, it had probably been dead for only a couple of hours. One of the biologists estimated that it was 6-8 weeks old. It had a fractured right humerus, femur, and tibia. It had a large hematoma on the left side of it's chest as too. On the inside, the diaphragm had ruptured, and the liver and stomach were in the chest cavity. Ouch. There was a lot of free blood in the chest and abdomen as well. That is probably what killed it.

I have seen that type of injury only once before, in special pathology class, but it was on a formalin fixed specimen, and it was really old. It was exciting to see it on a fresh specimen. This type of thing isn't uncommon in dogs that get hit by cars. If they don't loose a lot of blood right away they can live with it for a while, and sometimes it goes undiagnosed for months. Fortunately it can be fixed with surgery in those cases.


So I took a piece of spleen for the UAF study, and since it was so fresh, I took some blood from the heart for Kimberlee to do serology on. We were also curious to see what kind of parasites it had, so I took some feces and did a float. Surprisingly, it had a ton of coccidia, and some hookworm eggs as well. In this picture, the coccidia are the little protozoa, and the hookworm egg is the large egg in the lower right corner. It ended up being a very interesting necropsy.

After that I washed another abomasum, and I got to use a brand new dissecting microscope to look for worms in it. I didn't get very far though, because we had another necropsy specimen come in.

This one was a little brown bat. It was found in McGrath, a bush town in the interior of Alaska. It weighed only 8 grams, and Kimberlee necropsied it using a dissecting scope. It looked like it had hemorrhagic colitis, a portion of the colon was bright red and bleeding. We also both agreed that it was a female, and might have been pregnant. I have a picture of the tiny critter, I'll post it in tomorrows entry. So we froze the whole bat to send to the University of Alaska Museum, and we fixed the viscera in formalin to send to a pathologist.

I guess that's it. It was a good day, we had two necropsies.

Thursday, June 22, 2006

 

Solstice

6/21/2006

Today is the longest day of the year. Here in Fairbanks the sun will be up for 21 hours and 50 minutes (but it will be light for 24 hours). There is a big festival downtown called the Midnight Sun Festival, there's lots of food, music, and vendors. That had to wait until after work though.

I started the day doing fecals again (see left). It was the same as usual, some larvae, some fluke eggs, and some tape worm eggs.

The Tularemia case made the newspaper as well (the Fairbanks Daily News-Miner), so that was kind of exciting. Kimberlee is quoted in it several times, and they had some info about the disease for the public.

After I finished the fecals I necropsied a raven that someone reported was dead in a woodpile. I was planning on taking brain samples for West Nile Virus, but unfortunately the bird was pretty decomposed, so I was unable to. But I was able to figure out what happened to it. It had a fractured humerus and scapula, a dislocated stifle (knee), and the front 1/3 of it's beak was broken off. It was found kinda near a road, so it was most likely hit by a car.

After that I worked on abomasums, and found some more Ostertagia.


When I was done working, I headed downtown for the festivities. I ate some pizza, listened to some music, and bought a couple of souveniers. I think that the whole city was there, it was crowded!

At about 9:00 I headed to the ballpark with a few friends from Fish and Game to watch the 101st Midnight Sun Baseball Game. It was between the Alaska Goldpanners and the Beatrice (Nebraska) Bruins (minor leage teams).

The game started at 10:30, and it was a ton of fun! The game is unique in that it's so late at night, but they don't use artificial lights. The picture to the right was taken at midnight during the 8th inning.

It was a great game, and the Panners won 2-1 in the 10th inning with a line drive to the outfield. The game ended at 12:30 and it was still light out. It's strange being up here in the summer. It hasn't been dark since I got here almost 4 weeks ago. Last summer I had a real hard time sleeping for the first couple of weeks, but I'm pretty used to it now.

 

The Mystery Deepens


6/20/2006

I couldn't find an appropriate picture for today, so I put this one up that Nikki took on the peninsula. It's of a swallow sitting on the Beaver. I think it's cute.

This morning, Don (the biologist involved with the Eielson bear case), got the head out to investigate further. He was planning on just pulling a tooth for aging and measuring the skull. When he started skinning in order to measure it we noticed a lot of bruising over the skull, as well as a puncture hole in it. There were no punctures at all in the skin, so we never would have noticed it if he hadn't skinned it. So I went and got Kimberlee, and she decided to open the skull to have a look at the brain. When she cut the muscle off of the skull in order to open it, we noticed another puncture mark, only 3.5 cm away from the first (again, with no puncture in the skin). There really wasn't much damage to the brain though, just a tiny bit of hemorrhage, the puctures were pretty superficial.

We had been suspecting that another bear killed the yearling, but the punctures on the skull were only 3.5 cm apart, which is way to small for a bear (it's even too small for a black bear). So we were thinking of all these crazy scenarios involving a bear and a wolf, or two bears (an adult and a cub), or a pack of dogs or something. Finally we decided that the puntures were probably the upper and lower incisors of a large bear (rather than both uppers as we originally thought). The only thing that still seemed odd was that two bears were able to get into that munitions dump at the same time. This just goes to show that there is always something exciting happening in the field of wildlife medicine!

When we were finished dealing with the bear I was back to my usual stuff. I washed another abomasum and looked for worms.

Wednesday, June 21, 2006

 

Mystery Bear

6/19/2006

Well it's Monday of my last week in Alaska. I went camping and fishing over the weekend at a place called Tangle Lakes. It's a couple hours drive south of Fairbanks. While I was there I got this great shot of a cow moose and calf. They were crossing a narrow part of the lake together.


Now it's back to work though. This morning I washed another abomasum and picked the Ostertagia out of it, luckily it was a small one and it didn't take too long. Then I entered data for a little while.


Then the mystery started. The area biologist received a call from someone at Eielson Air Force Base that they had a dead bear. So Kimberlee and the biologist went to pick it up. Turns out it was in their munitions dump, which is a very secure area. They weren't sure how it had gotten in there or how it had died, they were thinking there was some foul play involved (like poaching or something). So we decided to figure it out.

It was a grizzly bear, but not a very big one (the biologist estimated it to be a yearling based on dentition). Here's a picture of me beginning the necropsy by measuring the length of it. It was pretty rotten too, we think it was sitting there for 2 days at least (it smelled BAD).

It had a large wound on it's back, right beneath the ribs, which kind of looked like the exit wound of a bullet. We couldn't find an entrance wound though (or any other external abnormalities). So we skinned it. There was extensive bruising all throughout the back musculature, as well as some severe bruising on both thighs and one knee. What appeared to be an exit wound didn't really fit the bill as we explored more, it didn't go anywhere, and there wasn't really very much hemorrhage around it. The viscera was pretty rotten, but it didn't look like a bullet had traveled through the abdomen or chest. It also had a shattered vertebra (L3 to be precise), but it looked like it had happened post mortem.

So we came to the conclusion that a larger bear had probably killed it, and then after it died it must have picked it up and shaken it (breaking the spine). That doesn't really make sense either though, since it's such a secure area, it seems odd that two bears were able to get in there at the same time with nobody noticing.

It was getting late at that point, so we cleaned up. But we saved the head so that a biologist can pull a tooth tomorrow (to age it) and measure the skull (they keep track of all that stuff). Kimberlee and I will also have a look at it to see if there is any evidence of head trauma.


 

Ostertagia


6/16/2006

This morning I started by entering the differential data that I collected yesterday. We have quite the Excel file going with all of the results of the bloodwork, fecal floats, larvae counts, and tissues collected.

The rest of the day pretty much all I did was sift through the abomasal (stomach) contents that I started yesterday. There was about 1 L of contents retrieved from it, which Nikki and I went through a couple of mLs at a time under a dissecting microscope. The worms that we are looking for (Ostertagia gruehneri) are tiny, they are thinner than a hair, red to brown in color, and about a centimeter long. The picture here is one I took of the clasping end of a male worm, it's magnified 100X. You can just barely see them with your naked eye. Using a dissecting scope makes it much easier.

Kimberlee also got the PCR results back from the hare spleen, it was positive for Tularemia (which we were expecting).

I guess that's about it. Those abomasums tend to eat up a whole day. We found a lot of worms though, we are going to count them, and they will be sent off to a parasitologist to confirm the species.

Tuesday, June 20, 2006

 

Muskox

6/15/2006

This morning I started doing differential white cell counts on all of the NAP and Mulchatna smears that we made in the field. They weren't too remarkable, there were two animals with a lymphocytosis (too many lymphocytes, which could indicate a viral infection), and most of them had an eosinophilia (which indicates parasitic infection, which we were expecting).

The culture results from the University of Alaska on the spleen from the hare came back as a strong positive for Tularemia. Fortunately we were careful handling it, and we put it in a biohazard bag afterwards to keep anyone from contacting the carcass. We are still waiting on the PCR results from the state public health lab, but we are expecting it to come back positive.

We also necropsied the muskox calf that I mentioned a couple of weeks ago. (the one that died up on the north slope) There's a picture of it below. It was a tiny calf, less than 30 pounds.

There were no signs of external trauma (just as we were told by the biologist), and it was in decent condition (not emaciated). We first opened the joints, since there have been a couple of animals positive for Chlamydia on the north slope (which can cause an arthritis). The cartilage looked a little eroded in a few areas, and the joint fluid was thick and red, meaning there was an arthritis going on.

The lungs looked like there was possibly some mild pneumonia with pulmonary edema, but it was hard to say for sure, those kinds of things can be post mortem artifacts.

At this point we were thinking it was Chlamydia, but when we started poking around in the guts we got our answer! Any guesses? It had an intussusception. An intussusception is an intestinal event where a portion of the intestine telescopes in on itself. That causes the blood supply to be cut off and the intestinal wall dies. When the intestine is dying the wall becomes permeable to all sorts of things, including bacteria, which could have spread to distant places in the body (like the joints), causing arthritis.

Intussusceptions (say that 3 times fast), are supposed to be extremely painful, and can lead to death within a day or two. Kimberlee informed me that they have been linked with coccidial infections in muskoxen, so I did a fecal float, but I found no coccidia. When we opened the rumen though, we were surprised to see that it was almost full of it's mothers hair. The hair could have caused a blockage in the small intestines which could have lead to the intussusception.

We don't know why it was eating hair though. One reason would be if the mother wasn't producing milk. But the animal was in good condition, and it had curdled milk in it's abomasum, so it was getting milk. We don't have a good explanation for why it was eating hair.

After all that excitement, I washed another abomasum and started picking Ostertagia out of it. I didn't finish it though, I will work on it again tomorrow.


 

Tularemia 101

6/14/2006

Here's a picture of some Francisella tularensis (the bacteria that causes Tularemia) colonies on an agar plate. The disease can cause people to have a fever, chills, headaches, diarrhea, muscle aches, joint pain, a dry cough, or progressive weakness. The CDC also lists it as a potential bioterrorism agent.

I necropsied the hare that I picked up last night, and it definately looked like it had Tularemia. The spleen was gigantic, and the liver was very large too. There were also no signs of trauma, indicating that it had probably died of disease. We submitted spleen and liver samples to the University of Alaska Fairbanks bacteriology lab, as well as to the state public health lab in Anchorage for a Tularemia PCR (polymerase chain reaction - a test that detects the bacterial DNA in a sample). We also took a few histology samples and fixed them in formalin so that a pathologist can interpret them later.

When I was done with that I finished up the last of the NAP fecals. There were 3 random samples that had been found near calf mortalities, as well as the calf fecals to finish. One of the random samples was loaded with Ostertagia larvae, some of them were still alive and squirming around under the microscope. None of the calves had any eggs in their feces, which makes sense, none of them had lived long enough to develop a parasite infection.

When I finished those I got all of the blood samples together that I will be taking back to Illinois to run cholinesterase assays on. I've already run 25 samples (before I came up here), and I will be bringing 78 more frozen samples back with me.

That's all for today. Hopefully I don't have Tularemia now.

 

Crunch Time

6/13/06

Here's a picture of one of three arctic ground squirrels that lived at camp. They had burrows through the tundra and would pop out to check out what was going on. They were very brazen, and would come really close if you gave them some food (cheetos for example). I'm not sure which one this is, but the guys at camp had named each of them, there's Francis, Justin, and Ron.

Today was another data collection day. I began by entering all of the fecal results from yesterday into a database. Next I went with Nikki to Alaska Air Cargo to pick up the gear that we had shipped from King Salmon (necropsy equipment, formalin fixed samples, frozen tissues etc.).

Then I started thawing a NAP caribou abomasum from last October (the 4th "true" stomach of a ruminant). Once it was thawed, I emptied all of the contents, washed the lining thoroughly, and sifted through the contents for adult Ostertagia worms. I didn't find any in this animal, but that was expected, it died in October, which is when the larvae encyst in the abomasal wall and go dormant until the summer.

That was all I did during the day. I was home making dinner when Kimberlee called to ask me if I wanted to go collect some specimens for a disease outbreak investigation. I of course said yes.

So a family in the nearby town of North Pole had called to report a "field of dead rabbits" near their home. I went out to investigate with a Fish and Game biologist. We arrived to find that there was no field, but there were 2 dead rabbits (varrying hares, actually) in the woods. One was too rotten to use, but I took the other one back to ADFG. The people who called reported that there had been several other dead rabbits in the area recently.

The primary thing that Kimberlee and I were worried about was Tularemia, a zoonotic disease that periodically shows up in rabbits and small rodents up here. It is a serious disease, and can potentially be fatal to people. So tomorrow we will necropsy that hare and submit samples for Tularemia diagnostics.

Monday, June 19, 2006

 

Back to Work

6/12/2006

Today I started working on the samples we brought back from the field. I began with the most fun of them - the fecal floatations! I did floats on all of the NAP and Mulchatna fecal samples that we brought back.

I found all sorts of interesting things. Below is a picture of an embryonated strongylid egg from one of the NAP caribou (most likely Ostertagia). The little worm was squirming around in the egg just waiting to hatch out. I think it's pretty exciting.


I also found Tricurid-looking eggs, Ostertagia eggs, Fasciola-looking eggs, a few Ascarid eggs, and some Coccidia. There were also some hatched out strongyle larvae in one sample (also probably Ostertagia). Every sample but one had something in it.

The ones that look like Fasciola (liver fluke) were very interesting to me. I didn't realize that there were liver flukes in Alaska. I asked Kimberlee about it, and she said that other people have seen eggs in the feces, but nobody has found actual flukes in livers yet. Weird.

I guess that's all I did today. It's not as exciting as being out on the tundra, but it's very important, and this is where all of the data starts to come together and we can start learning things about the NAP caribou.


 

Back to Civilization

6/11/2006

We stayed the night at the Fish and Game bunkhouse in King Salmon. There was no wind blowing our tents away, and no runing to the outhouse at 2am when it's 35 degrees outside.

Kimberlee, Nikki, Troy, and I went to the King Ko for breakfast. I had Russian French toast, which is regular French toast stuffed with cream cheese and topped with whipped cream (it's very healthy). I don't know if it's really what people eat for breakfast in Russia, but it was tasty.

Then we went with Troy to the hangar and helped him push the helicopter outside (he had to fly it back to Fairbanks today). Then we went to the airport and checked in for our flight. It's a teeny little airport in KS, there's no security or metal detectors or lines or anything.

After we were all checked in, Kimberlee checked out a dog owned by a Fish and Game employee. There's no permanent veterinarian in KS, normally a vet flies in twice a year to see appointments. So when people get wind that there's a vet in town they take advantage! Anyway, this dog had cut one of his pads deeply while at the beach, the doctor in town cleaned it out, glued it together with tissue glue, and bandaged it. The owner just wanted to make sure that it looked okay. It actually looked pretty good, so Kimberlee rebandaged it and told the owner to keep it clean and keep an eye on it.

Our flight left KS at 11:15, and we got to Anchorage at 12:30 only to find out that our flight to Fairbanks was overbooked and we might not have seats (argh! Why do airlines do that!?). But at the last minute we did end up getting seats. We arrived safely in Fairbanks at about 3:30 pm.

So now I'm here, unpacking my stuff and doing laundry. Tomorrow I'll go back to Fish and Game start working on all the samples we collected.

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