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how do you do a smear? do you just smear it on the slide or is it more complicated?
 

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Discussion Starter #23
Arklier said:
Decent microscopes are expensive. One that's not a toy will run you several hundred dollars. If you're only going to be doing a few fecals, then it's better to get the vet to do it.
"Decent" microscopes will only run you $60-$120 used on ebay. If you know what you are looking for, you can get something that is far more than "decent" for that price.

You need a 'scope that will run somewhere between 40 and 400 x, preferably up to 1000 x . Almost all 'scopes will have a 10 x objective on the eyepieces, then will have multiple (usually 3) objectives below running at 40 x, etc. To get the actual magnification, you multiply the eyepiece objective power by the lower objective power (so using a 40 x objective with a 10 x eyepiece is 400 x = 400 power magnification). If you stick with decent brand names or old college equipment, you should do just fine.

I know in an earlier post I called them dissecting 'scopes, because we used 'scopes like these in a histology lab in college after dissecting tissue. However, the common dissecting scopes don't have nearly the power that are required to see the protozoans and other parasites you will be looking for.
 

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Homer,

Do you know of any good 'how to' resources on the web about how to do fecals? Or any publications?

I have heard of some how tos from a couple sources, but thought it might be a good time to revisit this thread.

Thanks,

Melis

Homer said:
Arklier said:
Decent microscopes are expensive. One that's not a toy will run you several hundred dollars. If you're only going to be doing a few fecals, then it's better to get the vet to do it.
"Decent" microscopes will only run you $60-$120 used on ebay. If you know what you are looking for, you can get something that is far more than "decent" for that price.

You need a 'scope that will run somewhere between 40 and 400 x, preferably up to 1000 x . Almost all 'scopes will have a 10 x objective on the eyepieces, then will have multiple (usually 3) objectives below running at 40 x, etc. To get the actual magnification, you multiply the eyepiece objective power by the lower objective power (so using a 40 x objective with a 10 x eyepiece is 400 x = 400 power magnification). If you stick with decent brand names or old college equipment, you should do just fine.

I know in an earlier post I called them dissecting 'scopes, because we used 'scopes like these in a histology lab in college after dissecting tissue. However, the common dissecting scopes don't have nearly the power that are required to see the protozoans and other parasites you will be looking for.
 

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Discussion Starter #25
Check my first post. Both of those sites give good instruction, with the second one giving step-by step instructions on doing a float. My vet indicated that you are more likely to find some smaller organisms on a smear (talked about in this thread), but he runs both.
 

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I just finished examinig a fecal smear from my RETF that just began exhibiting the "neon spots". Anyway, I found many, many, many ciliated organisms; one that cruised around like an inchworm and was impossible to photo; and several nematodes which I believe are Rhabdias sp. Here they are, all images were taken at 40X. The ciliates are next to the measure bar.






 

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When this topic was started in 2004, it was mentioned that the majority of frogs had parasites (90%, with 80% more than 1 type)
What is being found now? Still the same prevalance in CB PDFs?


I also wanted to bump this really cool topic.
 

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tzen said:
When this topic was started in 2004, it was mentioned that the majority of frogs had parasites (90%, with 80% more than 1 type)
What is being found now? Still the same prevalance in CB PDFs?


I also wanted to bump this really cool topic.
http://www.dendroboard.com/phpBB2/viewtopic.php?t=33914&postdays=0&postorder=asc&start=30

Actual numbers start on page two. And as stated a few years ago in this thread, it is not hard at all to buy a cheap scope and find stuff floating around. Knowing what you are looking at and knowing what to do next is the important/harder step.

Rich
 

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Check out my photos in the gallery. I have an album of frog parasites. I have done hundreds of fecals, not only on my own frogs, but also on wild reptiles and amphibians. Here are a few observations regarding my limited experience with amphibians:
1. Wild caught frogs and toads are litterally "bag-o-worms". A heavy parasite load involving multiple phyla of organisms is the norm.
2. In captive bred frogs the infection rate is probably close to 100% if you count all forms of protista. Infection involving nematodes is also very high but do not involve anywhere near the diversity of different parasite species you'll encounter in wild frogs.
3. As pointed out earlier, flotation techniques are not nearly as effective as a direct smear with the small volume of feces you get from dart frogs. Most of the smaller protista are destroyed by the rapid osmotic change and are missed. Likewise the number of shed larvae by most nematode species can vary and false negatives are very common. For this reason I prefer to use a dissecting scope for an initial scan. In a watch glass or petri the entire fecal pellet can be teased apart and flooded with several drops of saline (.6%). Using the upper magnification levels (40X) on most dissecting scopes will immediately reveal movement. Most samples will litterally be teaming with ciliates and flagellates along with 1st stage nematode larvae. Some blastomere or larvated eggs are also often encountered. Further detailed examination can be done using a compound scope at much high magnification. I will often use a micropipette to lift specimens from under the dissecting scope in order to prepare a wet mount for much higher magnification.
4. Beware of "artifacts" that look like parasites or eggs. Fruit fly eggs, insect parts, pollen, etc have fooled many "experts".

Happy Hunting! There is a whole new world inside that little dab of fex.
(by the way: "feces" is plural for fex"

George
 

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For those doing smears, are you not even using a drop or two of flotation fluid or some type of liquid? If so what type of fluid? My smears with fresh fecal samples are coming out a bit dry, and some smear tutorials have mentioned using 1-2 drops of fluid on the smear then placing a cover slip on top. Any advice is appreciated, thanks!
 

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Okay, well at my work we have a .9% saline solution. Which if you wanted to make that, it'd be 9 grams of NaCl per liter of water.

If you wanted to figure that out yourself, it'd be .9% solution = [9g (or mL) of NaCl/1000mL H2O]x 100

Or someone correct me if I'm wrong, but that's how I figure it :)
 

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Re:

lacerta said:
Check out my photos in the gallery. I have an album of frog parasites. I have done hundreds of fecals, not only on my own frogs, but also on wild reptiles and amphibians. Here are a few observations regarding my limited experience with amphibians:
1. Wild caught frogs and toads are litterally "bag-o-worms". A heavy parasite load involving multiple phyla of organisms is the norm.
2. In captive bred frogs the infection rate is probably close to 100% if you count all forms of protista. Infection involving nematodes is also very high but do not involve anywhere near the diversity of different parasite species you'll encounter in wild frogs.
3. As pointed out earlier, flotation techniques are not nearly as effective as a direct smear with the small volume of feces you get from dart frogs. Most of the smaller protista are destroyed by the rapid osmotic change and are missed. Likewise the number of shed larvae by most nematode species can vary and false negatives are very common. For this reason I prefer to use a dissecting scope for an initial scan. In a watch glass or petri the entire fecal pellet can be teased apart and flooded with several drops of saline (.6%). Using the upper magnification levels (40X) on most dissecting scopes will immediately reveal movement. Most samples will litterally be teaming with ciliates and flagellates along with 1st stage nematode larvae. Some blastomere or larvated eggs are also often encountered. Further detailed examination can be done using a compound scope at much high magnification. I will often use a micropipette to lift specimens from under the dissecting scope in order to prepare a wet mount for much higher magnification.
4. Beware of "artifacts" that look like parasites or eggs. Fruit fly eggs, insect parts, pollen, etc have fooled many "experts".

Happy Hunting! There is a whole new world inside that little dab of fex.
(by the way: "feces" is plural for fex"

George
A couple thoughts.
George,
Your #s don't quite jibe with the thousands of Dart Frog fecals my brother has run and followed up on. I have to wonder why a vet would say that almost 100% of CB would be infected when almost 100% of newly morphed froglets are not. Does this 100% stat include your frogs? While it is much more difficult to raise a clean froglet in an eggfeeder environment , quarantine (with proper dis-infection of other things going into the vivs) can and should wipe out pretty much all of the nasties before introduction into a clean viv. Therefore having a clean viv + clean adults should= clean babies. How is it possible that all of the froglets being produced are infected? I know mine are not.
Also, out of the hundreds (and hundreds, and...possibly thousands) of strickly WC Dart Frog fecals my brother has run he has found most WCs to be much less infected than the average CB that has been walking around in it's own un-quarantined, nematode/proto/whatever poop day after day after day. This should not be hard to grasp.
And, it should be stressed , as I have already, that it is not hard to set up a fecal (I have done hundreds myself) , but IDing properly the eggs/worms/whatever is. Very hard at times. And after you do possibly ID whatever a consult with a vet is needed to properly treat (or not treat if you choose) and understand the issues at hand.

Rich
 

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Rich, 100% infection rates are the norm if one includes both ciliate and flagellate protistans. Most do not show up in fecal flotations as the steep osmotic gradient will literrally burst them apart. Short of raising our frogs on a sterile substrate medium (impractical IMO), frogs will quickly develop a parasite load because many of these organisms are faculative parasites that are both freeliving soil/water denizens capable of taking residence in the g.i. tract of metazoans if the opportunity presents itself. Colpoda is just one of many examples that come to mind. It is found in both water and soil and is capable of encysting itself in dry soil or on plants. In setting up a vivarium with sphagnum, or other natural substrates it is hard to imagine that we would not be introducing many of these organisms. But most of these critters are harmless and part of the normal intestinal biota found in all frogs. Remember, parasite infection is not synonymous with a disease condition, and prophylaxis and treatment should be reserved for only those parasites that present a significant health risk.
George
 

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George,
I get a bit worried when a professional writes a post and does not talk down to the level of the crowd reading. The whole "WC bag of worms" and "100% parasitic infection rate" is just not the case in Dart frogs . Parasite infections can be at best neutral to the frog and at worst fatal. A parasite by definition (most people's) does no good for the host. If it does it is not a parasite. There are in-fact many protozoa that do no harm and will be very easily confused with protozoa such as coccidia by the first time/home set-up/not used to IDing Dart Frog parasites crowd that this thread is addressing. But protozoa with no potential for harming the host and not really leaching off the frog are not what I call parasites. And it is not what people should be running fecals to find.Or be too concerned about for that matter.
As far as the WCs having many/more parasites , be they worms or not , it is just not what we have found to be the case in many, many tests in the past five years.
I propose a test or five. I would love to send you a few fecals to have you run. I can label them 1-5 or a-e or whatever, you read them and take some pics and I will post the frog and the situation of each (to a mod ahead of time , if you like). I would be very interested to see if there are in-fact parasites in each sample.
I would like to say that the main reason we do fecals is to find out the state of health of our frogs pertaining to parasites. If the frogs have parasites there are options and it is my belief that all parasites that can be cleaned out should be. Not all protozoa. The three biggies (parasites)I want to wipe out are hook, and lungworms and coccidia. Coccidia not being curable but treatable. Many of the other protozoa and I suppose maybe, maybe even a worm species or two may be just fine in our Darts' systems, but the 100% thing and bag o' worms makes it sound as if they have 'em coming into this world and they will have them going out and that is the way it is. Period. ...to the average person reading your last couple posts.
Let me know if you are willing to run my fecals. I am very interested in what your finding may show.

Rich
 

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I must also state that Dart frogs are unique in the fact that they are the only amphibian adversely effected by coccidia, according to Wright and Whittaker. They also may be unique in the fact they have much less true parasites in the wild.

Rich
 

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Rich,

Did they state a reason why they have less true parasites? Maybe because of their toxicity? Also I know what you are talking about when you say you need a trained eye for spotting parasites. With all the different kinds of nematodes (which all basically look the same and don't all cause disease ex: plant nematodes) and normal intestinal fauna it is hard to differentiate between disease causing organisms.

Mike
 
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