I'm in need of a protozoan expert.

[13mur 6]

Hey all,

Here's what happened. My male snake who's been sick since December has gone through 2 doses of flagyl, 2 doses of panacur, and 10 doses of baytril during a time period of 7 months. He's no longer regurging, and he's eating properly, it's just that he's still a skinny mofo when my blizzard who eats half as much as he does is twice his size and 6 times as heavy and they were hatched on the same day.

The vet that I went to said the fecal sample I gave her was clean.

I did two of my own fecals only going as far as 400x magnification and found them to be clean also.

However, today, I ditched the dye, ditched the phase contrast and just used saline and fecal smear, looked at it at 1000x magnification using one hell of a scope (full plan optics, electronically controlled everything), and found millions upon millions of tiny tiny gyrating globular things. These were seriously tiny even at 1000x, about the size of the comma on my keyboard. They didn't move much in terms of distance at all. They all sorta stayed in a spot and twitched/gyrated around that spot. I couldn't get the scope to focus on the moving things too well and no greater magnification was available (but 1000x is pretty damn high to begin with). I tried manuall focusing, but they were still kinda blurred, but I could make out the general shape of them and most of them were spherical and once in a while there was one that was sort of oblong.

I've also found what seems to be an egg. It was a bit bigger than the protozoans themselves, sort of squashed hexagonal shaped with a oval dark spot in the middle that had a dark line through it.

I'm not going back to my vet since she's only told me lies so far and giving me bogus drugs (I don't want marshmallow flavored drugs, I just want the damn drugs at the correct doses). And she's too easy to anger, who the hell does she think she is yelling at me for not feeding my snake EVERYDAY? Who the hell feeds their snake everyday?

Anyone chance to take a guess at what this protozoan may be? I have a feeling it might be coccidia from the descriptions I've read online. I'll try to take a few digital pics of these things and post them if that would help (this scope is one monster of a scope, does everything). I have a copy of Klingenberg's book, but I don't see anything like it mentioned in the book (also the book doesn't describe the behavior of protozoal movement).

Also, this must be the 5th time I'm asking this but could any of you recommend a reptile vet in the NYC area? That would help me a WHOLE lot.

-13mur 6

[Gregg]

www.arav.org/

or

www.herpvetconnection.com/

[CornCrazy]

You shouldn't have to put the scope on 1000x to see coccidia.
I'll find out some info at work...one of our doctor's used to have some contact info on a good parisitologist. You may be able to contact him and see if he could take a look at a fecal sample from the snake. I won't be at work until tomorrow, and I'm not sure if that doctor works then, so I will get the info as soon as possible.

Here is a picture showing the oocysts of some Isospora species.

[13mur 6]

Does Coccidia have a motile stage? I thought Oocyst didn't move, what magnification where those images taken at?

Also, in a lot of the life cycle drawings online I couldn't find anywhere where it said that actual protozoan stage. Isn't coccidia a protozoan? The slide was absolutely swarming with the small tiny globular protozoans and I didn't even notice they were they're at 400x.

I'll look for some sodium nitrate so I can do a float next time, I think I'll call a couple vets today see if anyone can help, or ID what I found.

-13mur 6

[CowBoyWay]

"Parasitology Images List ...
possibly what your looking for...

Here is a collection of images of medically important parasites.
The majority of the images show diagnostic stages of the parasite concerned, in the most commonly encountered clinical specimen.
Each has the name and stage of the parasite shown, along with an approximate scale bar for comparison."
http://www.life.sci.qut.edu.au/LIFESCI/darben/paramast.htm

Parasitology-related links
Cryptosporidium, coccidia, and other parasitology-related pages...
http://www.ksu.edu/parasitology/links

http://biology.unm.edu/biology/coccidia/home.html

[carol]

How long has it been since he has stopped regurging? It may just take some time before he is back to normal. If your vet is giving you advice to feed the snake everyday, I would definately go to a new vet. I don't know any in your area, but the best thing to do is call around and ask questions... like how long they have been in practice and if they have references. Being in New York you should be able to find a vet that specializes in reptiles instead of "doing them on the side". Don't panic too much, those little things in his poo could very well be something that is sappossed to be there. I don't recall... did you ever get a culture done?
Hopefully CornCrazy can help you out because it really sounds like your vet is not very knowledgable in reptiles.

[13mur 6]

It's been months since her last successive chain of regurges. The very last regurge was about 2 weeks ago and that was from me feeding her a lil too much, and he does seem to be recoverings somewhat, but not really (more like remaining thin, but being more active, inquisitive, responsive, etc).

I found one vet near my place that charges $18 for just a smear (I don't know why the heck NYC people have to seprate the fecal test like that, they charge like ~$20 for smear and then another ~40 for a float, and they refuse to accept my prepared fixated and stained slides of the thing). I couldn't get numbers for the vets and I had to scrounge for them and only found one (there's only 3 reptile vets listed on the ARAV site in the NYC area).

I did get a bacteria culture done but that was a couple months ago, and all that was found was a strain of Proteus and Salmonella, and baytril was given to try to rid the proteus.

I am sort of questioning what exactly it was that I saw, I used the wrong oil (actually it wasn't even oil it was a mislabeled bottle of acetone), and I couldn't focus on the things too well. I'll run my own fecal again, take some pics, maybe even movie of the things, save them on a disk and print the pics, take whatever's left of the poo and give the package to the vet to figure out if his scope doesn't have the snuff to see the itty bitty things I saw. Along with that I'll try to find someone with some gram stains and look for any bacteria that aren't supposed to be there.

However I still do think it's coccidia my snake has, I just need to find some cysts in the next fecal to be sure (not one person anywhere mentions what the motile stage of the coccidia protozoan looks like or behaves, but I bet that's what I'm seeing), the symptoms match almost exactly to other people who've had coccidia in their snakes. Also, from what I gathered from looking at all the info online, seems like there are male/female stages and one is sort of globular and one is sort of sausage shaped (what I saw).

Hey corncrazy, what power should I use to view cysts? 400x? Any common dyes you recommend? Meth blue work? or is it best with no dye? Someone explain acid fast staining to me and how it works.

-13mur 6

[CornCrazy]

Ummm....I don't remember right off the top of my head which power we can view the oocysts with. I do know that the microscope at the vet where I work only goes up to the 100X (oil immersion) objective, and we can see them on it.

I don't know for sure if coccidia has a trophozoite (motile) stage or not. I've never seen anything except the oocysts. Giardia does have a trophozoite stage, though. I'm going to post a picture of Giardia oocysts and trophozoite stage. I had to turn the picture sideways in order for it to upload. Please let me know if what you saw resembles them. Oh yeah - The book I got these pictures from doesn't tell what magnification they were taken at. Not very helpful, huh?

As for dyes...we only have gram stains, dif-quick stains, meth blue, and a fecal stain (which I've never used before). I have never actually stained a fecal before so I really won't be any help in that area.

Sorry that I'm not much help right now. I will try to find out some more info, or at least get some contact info for you. I'm leaning toward what you saw as being some kind of bacteria (good or bad?) instead of a protozoan. I can't think of anything else that you might have to use such a high power objective to see.

[13mur 6]

Hey corncrazy, you see those clearish globules in the background of those pics you just posted? THAT'S what it looked like at about that size too on the 100x obj., and they were sort of off focus like the way it was in the pic and it was as best as I could focus on them. And on the middle pic see the longish one right above the bottom cyst? I've seen a few of those too. They moved in a twitchy motion and sort of gyrated around one spot (they were non-progressive motile) and they were about as concentrated as the left most pic shows. Might be acidophillus + E. coli if it's bacteria, since I've been feeding my snake acidophillus supplement every meal. But the gram stain should show that next time I do a fecal. But I still do suspect something is wrong since his poo doesn't look normal.

From what I've read, there's a microgamete that actively enter epitheleal cells containing macrogametes during the sexual cycle (actively through movement?). There's also merozoites that are released in the asexual cycle, and there's also a sporozoite on top of that out from the sporocysts that are released during the infectious stage of the oocyte that also actively enter cells. Bloody parasitologists people always using this ambiguous terminology.

Thanks for the pics, what book did you get them from?

-13mur 6

[CornCrazy]

You said on the 100X objective? Do you mean the 1000X objective as posted before? I'm not 100% sure what the clear things are...fat sometimes look like that. We see lots of animal who are given table food, and they ALWAYS have fat in their fecals. In the middle picture, the long item is probably a rod , which is a bacteria. That's the only thing I have ever seen which is that shape.

I want you to please understand that I am not a doctor or a parisitologist, so please don't treat your snake based on any info I may be able to dig up for you. I cannot make a diagnosis...
I can give you pictures, contact info, and etc. Ultimately you are going to have to find a vet to make a diagnosis. I know that you aren't having any luck with that where you live, though. There is an avian lab that does testing on reptile feces as well. I do not know how expensive it is, but you may want to check into it. The website is: http://www.avianbiotech.com/

I got the pictures from a book called the "Pfizer Atlas of Veterinary Clinical Parasitology." It has an ISBN number, but I've not been successful in finding it available online or anywhere else. The number is ISBN 0-9678005-3-6. It is an excellent book for most of the common parasites and protozoans seen in a vet clinic.

[13mur 6]

Yep, I understand completely, I guess I like to report my findings, you never know when you might actually tag someone who's seen something similar, or knows a hella lot about what you've seen. Who knows, this thread my help someone who's overly curious and does everything him/herself 2 years down the line when he/she's found something like this in his/her snake feces.

I actually called a vet listed on the ARAV site not too far from where I live, though they make you pay through the nose for a comprehensive fecal, and I couldn't find the numbers for the other two vets (I got this vet's number from doing a search, and found his number on an avian vet listing, so I don't think he's a snake specialist, but we'll figure that out soon enough).

100x objective as in (100x objective) x (10x eye piece), so yep, 1000x maginification.

I don't think fat blobs move though, these things were moving quite vigorously, I'll try to scrounge for that book at my research lab's RARC (they're like the in house vets for us) office library. Thanks again for these lil tid bits of info.

-13mur 6

[CornCrazy]

100x objective as in (100x objective) x (10x eye piece), so yep, 1000x maginification.

That really bad...as long as I have worked at a vet's office, I was never taught that the 100x objective magnifies 1000x. I am currently in school to actually become a licensed vet tech, but I haven't had microbiology, yet. I guess I still have a lot to learn!

I'll try to get the info for the parisitologist for you. Maybe he can help.

[13mur 6]

Mm... that's not ALWAYS, the case, but I think the standard microscope (almost all lab microscopes) have a 10x eye piece. I think I own a set of 15x eye pieces somewhere, but I don't use them too often since I can't adjust diopeter settings with them and it makes my eyes hurt. But yeah, the magnification is defined as objective mag. multiplied by the eye piece mag.

-13mur 6

[Susan]

Those little round, oblong and even rod-shaped things that move around alot in one place are BACTERIA. That is also what you identified in the second set of pics posted for you by Corncrazy. They are perfectly normal in your snake's fecal sample. In fact, if you try to get rid of all of them, you will be doing more harm than good. Your snake needs good bacteria to help it digest it's food, which may be why your snake isn't gaining weight as it should. I would suggest that you pick up some BeneBac for reptiles or birds and give your snake several doses over several weeks. It will put good bacteria back into your snake's system.

And if this will help you feel better about my info, I'm a certified vet tech that has been practicing my profession for over 20 years. I have performed thousands of fecal exams over the years and can identify just about anything that can possibly be found in a fecal sample.

[CornCrazy]

I was hoping you'd post a reply here. I've been doing fecals for quite some time, but haven't actually taken a microbiology class so there are still LOTS of things I have to learn to recognize under the microscope.

13mur6 - The doctor I needed to talk to about the parasitologist didn't work Saturday. I won't see him until Tuesday. I'll try to get that info for you then.

[13mur 6]

Thanks Sue, that's just what I needed to hear. I think I was spazzing out about it because of the movement behavior of the lil things (I never knew bacteria moved around much at all).

And thank you CornCrazy for all of your efforts at helping me.

See, we did learn something through all this after all. Now to get a sample to this other vet to truly see if my snake is clean (Hopefully he will be as well versed in parisitology as your are Sue).

-Lemur 6

[Susan]

If you really want to get freaked out by movement under a microscope, you should see Spirochaetes! They're like bacteria, only just a bit longer, and shaped like a lightning bolt. They zip around a direct smear (feces mixed with water or saline) like Mario Andretti!

[13mur 6]

Hey everyone,

I did my fourth fecal today. This time I did a float using a sugar solution (20 min to be on the safe side, that stuff was thick) I can't say I found anything, I couldn't find anything bigger than a clump of bacteria other than the odd amorphous blob. The other things I saw were just some random artifacts (hair fragments, oil blobs, etc).

I did another smear as well, this time supervised by a doctor who's practice is microbiology (and a sweet guy too, likes to take people out for pizza). He agreed that the little blobs and rods are bacteria, and that if there were any cysts, I'd see them before I saw the bacteria. I am a bit wary though, since I did see things that resembled cysts, but he tells me they were too small to be cysts, but guess I have no one else to trust and his 40 years of experience dealing with human microbes is worth trusting IMO.

Also, can cysts be "squished" if I press too hard on the coverslip? Maybe that's why I'm not seeing anything.

I still have no luck at a vet. Today I've only seen vet techs who seem to make it their business to advertise their vet and claim their vet has "special interest in reptiles". BTW, how much should an acid fast stain run about? I'd be interested in learning how to do one myself (I bet I can find a kit in one of those big catalogs at work).

Oh, and my snake bit me. His feeding response is at an all time high. I hope he continues to go on like this and gain some friggin weight.

-Lemur 6

[panda]

Modified Acid-fast Stain Procedure from CDC
(Page could not be found today for some reason)

Reagents:
1. Absolute Alcohol
2. Acid Alcohol 10ml Sulduric acid + 90 ml absolute alcohol
3. Kinyoun's Carbol fuchsin
4. 3% Malachite green

Procedure:

1. Prepare a smear with 1 to 2 drops of specimen on the slide and dry on a slide warmer at 60 degrees C. Do not smar too thick
2. Fix with Absolute methanol for 30 seconds
3. Stain with Kinyoun's carbol fuchsin for 2 min.
rinse with distilled water
4. Destain with acid alcohol for 2 min rinse,
5. Countersatin with Malachite green for 2 min rinse briefly.
6. Dry on slide warmer. for 5 min
7. Examine 200 to 300 fields at 40X or higher objective. to confirm internal morphology, use 100x oil immersion.

Let me know what you find.

Mark