# How to run fecal samples



## Homer

For those of you with access to a decent 'scope, here are some good websites that should help you get an idea of the concepts involved in doing a fecal sample yourself.

http://www.microscope-microscope.org/ap ... alysis.htm

http://fiascofarm.com/goats/fecals.htm

While the animals on which the fecals are being run are different, the process and concepts are universal. Further, the genera of parasites that are of consequence tend to be readily identified by researching the appropriate texts. However, you can get a pretty good overview from some of the excellent pics available on many websites. With that info, almost anyone should be able to at least point their vet in the right direction, especially if they have a copy of the coveted amphibian husbandry book that keeps popping up. 

If you have a large collection, or just a proclivity to enjoy microbiology, there is no reason why you shouldn't be able to plunk down the money to buy a decent 'scope and do your own fecals. Once you've done 10, you've paid for your scope. 

I'm just curious as to how many other froggers out there do their own fecals? Give a shout out!


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## khoff

I got a microscope on the way, and I'm gonna try and see how it works out. Thanks for the great links.


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## addam4208283

I have a ton of information on this stuff because of my work I should post some pics. and other info; might help you guys out. 
Just so you guys know doing a fecal float on PDF crap won’t work to well, instead do smears, this will give you a better sample to look at. You would only do a float if you had something like a teaspoons worth of crap. With a smear all you need is a very small amount. I have done both methods with great results, but I think the smear works out a little better.
Hope this helps.  
ADAM


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## melissa68

*Thanks Homer!!!*

Thanks Homer for the info. 

Melis


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## Homer

Thanks, Adam! I was planning on running tests using both the smear and float methods to see which was more effective, but it sounds like you answered my question already. 

Let me ask you, in your experience, have you found that a vast number of captive born frogs have parasites? I have heard it said that this was true, but find it a questionable statement . . . particularly considering the source.


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## addam4208283

I first off want to say that I am not an expert on this subject. I have done probably about ten to fifteen frog fecals with another tech, and doctor at my work who are far more experienced in this then I. some of these fecals include both my frogs as well as a coworkers whites tree frog. But with that said about 85-90% of all the fecals done have had some sort of parasite in them. 

Also I have not forgotten about the pics. I promised. I have a few from a fecal I have done on my frogs last week but I am going to see if I can get some others pics like cocci. just so you guys can compare to. Unfortunately they will probably come from a dog/cat because my frogs did not have this as a parasite; but in any case at least it’s something that you guys can refer to.

i hope this helps.  
ADAM


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## Homer

Thanks for the info!


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## Homer

Do you know if the frogs you tested were cb or wc?


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## addam4208283

CB.
ADAM


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## addam4208283

Or at least that’s what i assumed them to be. Other then my coworkers and the four I have mainly the rest come from petco, and petsmart.
ADAM


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## Homer

While possible, I doubt that most animals from Petco or Petsmart are cb. Additionally, I can definitely see a good chance for cross-contamination due to their poor attention to cleaning tanks between use and their penchant for mixing animals (i.e. green anoles--usually wc--with tree frogs). 

Dr. Frye used to advertise that 80% of the frogs he tested had 3 or more parasites. That doesn't really get at my question, though. I'm trying to determine what the percentage of average cb frogs in the hobby have parasites--not the percentage of all frogs taken to a vet. Most people are going to get fecals when their frogs start behaving poorly. I would also venture to guess that nearly all (if not truly all) wc animals have parasites. So that muddies the statistics a bit (a lot, actually).

I was just wondering whether anyone was doing proactive fecals on their collections and what their experiences were with percentages of infected frogs. While it is intuitive that bacteria could be easily transmitted from parent to offspring, the infective mechanism for other parasites to be transmitted to offspring when the eggs are collected from a petri dish is not so obvious (unless feces are left on the petri dish). That leads me to believe that the rate of parasitically infected animals that are cb should be significantly lower than wc, and that comports with common reasoning in the pet hobby as a whole. I was just interested in seeing whether common reasoning bore out in reality.


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## melissa68

*Good questions*

Homer,

You make some really valid points. I don't think anyone in the df community doubts Dr. Frye's numbers, results or his findings, but you make a good point that many (not all) of the animals who were tested might have been showing other symptoms. 

Without someone putting up a post...."send us your froggie poop for a study..." it would be hard to gather good numbers. Plus, there are so many other factors to take into consideration as well. Protocols, supplies, ect would need to be created so each sample arriving to be tested would conform to basic criteria. It would be interesting to speak with other vets who treat zoological or other large collections to see what they have found. 

Melis







Homer said:


> While possible, I doubt that most animals from Petco or Petsmart are cb. Additionally, I can definitely see a good chance for cross-contamination due to their poor attention to cleaning tanks between use and their penchant for mixing animals (i.e. green anoles--usually wc--with tree frogs).
> 
> Dr. Frye used to advertise that 80% of the frogs he tested had 3 or more parasites. That doesn't really get at my question, though. I'm trying to determine what the percentage of average cb frogs in the hobby have parasites--not the percentage of all frogs taken to a vet. Most people are going to get fecals when their frogs start behaving poorly. I would also venture to guess that nearly all (if not truly all) wc animals have parasites. So that muddies the statistics a bit (a lot, actually).
> 
> I was just wondering whether anyone was doing proactive fecals on their collections and what their experiences were with percentages of infected frogs. While it is intuitive that bacteria could be easily transmitted from parent to offspring, the infective mechanism for other parasites to be transmitted to offspring when the eggs are collected from a petri dish is not so obvious (unless feces are left on the petri dish). That leads me to believe that the rate of parasitically infected animals that are cb should be significantly lower than wc, and that comports with common reasoning in the pet hobby as a whole. I was just interested in seeing whether common reasoning bore out in reality.


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## addam4208283

Here is a pic. of a nematode.










And another.


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## addam4208283

Here is a quick pic. of a ciliate; this pic. is taken under 40X.










I forgot to add that the nematode pics. were taken under 10X.

Hope this helps you guys out a little.
ADAM


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## addam4208283

Here is another pic. that I took, I am not 100% sure because of the quality of the pic. But I believe this is cocci.










Something else I forgot to mention above is that you can see both the ciliate, as well as the nematode moving around. The ciliate you notice movement less compared to the nematode.

hope this helps
ADAM


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## addam4208283

Cocci. was taken under 40X with min. oil.
ADAM


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## Guest

I am new to the list. I work as and RVT for a an exotic vet. I do numerous fecal exams daily on reptiles and birds, but not a lot of frogs. I just recently aquired some captive bred dart frogs, and have found larval worms in their stool that I strongly suspect are strongyloides. I have also found these parasites in a milk frog. 
Has anyone treated frogs with this parasite and had success? I am getting ready to put these animals in a planted vivarium and do not want to contaminate the tank and then have to replant it later. One of the other techs at our practice has been trying to clear her frogs on these worms with no success.


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## snmreptiles

*What kind of scope is needed?*

I just got on EBAY to look at prices of microscopes and am COMPLETELY lost!! I have wanted a microscope since getting the book "Understanding Reptile Parasites" by Roger Klingenberg! A GREAT BOOK btw. Anyway what am I looking for as far as options and others. 

Thanks,
MIKE


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## Arklier

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.


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## khoff

I disagree. You don't need anything too fancy run a basic fecal. I picked up a microscope that was a college lab's old equipment (off of ebay). It was like $50 and I have done fecals with it. The only hard part about fecals is identifying what you see. Hopefully the book you have will help with that. What you should be looking for is a basic compound microscope (just like the ones everybody used to use in high school). I wouldn't worry too much about picking up a used one, they are relatively easy to take apart and clean if needed (at least mine was). Hope this helps.

Kevin Hoff


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## Guest

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


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## addam4208283

I just take a Q tip and mash up the poop, then just roll it on the slide.
hope this helps.
ADAM


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## Homer

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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## melissa68

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.
Click to expand...


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## Homer

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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## melissa68

*other sources*

Did a quick search and found a decent explanation of different types of fecals to perform and how to prep the slides. 

The first two are the most commonly done: http://cal.vet.upenn.edu/dxendopar/techniques/comfecal.html

Melis


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## rmelancon

Since the "Fecals at IAD" discussions I've been interested in others who run their own fecals. Anyone have pix of things they have found, nematodes, ciliates, etc? Any methods you use to help identify the "bad bugs" from the "pieces of crap"?


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## bwebb

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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## tzen

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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## Rich Frye

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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## lacerta

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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## Corpus Callosum

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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## chibisan

I believe you would want to use a saline solution for the smear.


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## Corpus Callosum

Which recipe would you recommend for amphibian fecals? If I were to make my own solution.


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## chibisan

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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## Rich Frye

*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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## lacerta

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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## Rich Frye

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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## Rich Frye

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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## lamaster

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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## Ed

See Poynton, Sarah L.; Whitaker, Brent R.; 1994; Protozoa in poison dart frogs (Dendrobatidae): clinical assessment and identification; Journal of Zoo and Wildlife Medicine 25(1): 29-39 

They indicate that there was no difference in prevelance of infection of protozoa between wild caught D. auratus and frogs maintained subsequently in captivity and that most of the intenstional protozoal apprear to be commensuals rather than parasites. 

Ed


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## pl259

Hey Rich or anyone else,
How about posting a pic of a typical hookworm found in PDFs. They seem to be a pretty common parasite.


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## Rich Frye

lamaster said:


> 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


Mike, sinse it is endoparasites we are talking about I doubt it could be because of skin toxins. But I'm not 100% sure.
"They" would be me and my brother and I can only speculate that not walking around in their own infested crap all day in a ten gal may be one reason.
I have looked at a number of fecals and all I can say is I am very glad to have a brother with an experienced eye to look things over.

Rich

Rich


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## Rich Frye

Ed said:


> See Poynton, Sarah L.; Whitaker, Brent R.; 1994; Protozoa in poison dart frogs (Dendrobatidae): clinical assessment and identification; Journal of Zoo and Wildlife Medicine 25(1): 29-39
> 
> They indicate that there was no difference in prevelance of infection of protozoa between wild caught D. auratus and frogs maintained subsequently in captivity and that most of the intenstional protozoal apprear to be commensuals rather than parasites.
> 
> Ed


Ed, not sure why you bring this up. Was it to point out that that most/many protozoa are not issues (parasites) with darts , as we have agreed to. Or that non-parasites are about the same in the wild and captivity?

Rich


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## Rich Frye

pl259 said:


> Hey Rich or anyone else,
> How about posting a pic of a typical hookworm found in PDFs. They seem to be a pretty common parasite.


I do not have any. But my brother does. I ask the next time I talk to him.

Rich


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## Ed

Rich Frye said:


> Ed said:
> 
> 
> 
> See Poynton, Sarah L.; Whitaker, Brent R.; 1994; Protozoa in poison dart frogs (Dendrobatidae): clinical assessment and identification; Journal of Zoo and Wildlife Medicine 25(1): 29-39
> 
> They indicate that there was no difference in prevelance of infection of protozoa between wild caught D. auratus and frogs maintained subsequently in captivity and that most of the intenstional protozoal apprear to be commensuals rather than parasites.
> 
> Ed
> 
> 
> 
> Ed, not sure why you bring this up. Was it to point out that that most/many protozoa are not issues (parasites) with darts , as we have agreed to. Or that non-parasites are about the same in the wild and captivity?
> 
> Rich
Click to expand...

Actually both Rich.. 

Ed


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## lamaster

Rich, 

I know they hold keep the toxins in the skin. I was thinking since they make the toxins from a food source maybe there would be small amounts of toxins in the G.I. tract. I don't really know how the whole process works. I guess I am just wondering if they metabolize the toxins or just store the toxins they collect from the prey. I think same organisms like the Monarch butterfly just store the toxin and dont create it. I find it interesting so any information would be helpful thanks.

Mike


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## Rich Frye

Mike, 
I'm not a toxin expert , but I am fairly sure the toxins are restricted to the skin. It should not be tough to deduct that at least one very big reason CBs not properly quarantined would have more true parasites would be the fact they are walking in their own infested poop day in and day out. 
Rich


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## lamaster

Rich

I understand the whole walking in their feces all day. That is why I can wait to put my frogs into my vivs after a proper quarantine. Also, thanks with your input on the toxins I know there are very limited resources on the topic. 

Mike


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