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  #41 (permalink)  
Old 09-19-2019, 11:51 PM
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Default Re: Strange U shaped lump

Putting in formalin fixates the tissues, delaying the decomposition. Unless you have a microtome and wax fixation with H & E stain, fixating anything isn't going to be very helpful unless you send it to a lab.

It will probably cost a couple hundred dollars for IHC, or histopath. You really also need to know what your differential for cause of death is. If it is fungal, bacterial, etc. then fixating in formalin will not help you need fresh tissue.
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  #42 (permalink)  
Old 09-25-2019, 07:57 PM
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Quote:
Originally Posted by ghartman View Post
Putting in formalin fixates the tissues, delaying the decomposition. Unless you have a microtome and wax fixation with H & E stain, fixating anything isn't going to be very helpful unless you send it to a lab.

It will probably cost a couple hundred dollars for IHC, or histopath. You really also need to know what your differential for cause of death is. If it is fungal, bacterial, etc. then fixating in formalin will not help you need fresh tissue.
Then i wont bother with the formalin unless i find something so bizarre in the scapular area that its worth saving to share with others. It came in today so im likely going to go through with it today. I will just let it thaw and then go in with the tools i have. I may still collect samples for the slides even if nothing can be diagnosed from them. At the very least they could demonstrate what happens to tissues after freeze/thaw. This is something i personally find very interesting.
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  #43 (permalink)  
Old 09-25-2019, 08:01 PM
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Default Re: Strange U shaped lump

So whats going on with this?

I dont buy the need to mail/do/ histology/freezing bs. This is what I find disappointing about internet forums.

Here we had a clear opportunity to at least LOOK at something. Everyone knows that freezing destroys tissue character.

I think you found Bones with helminths found or otherwise visual through collapsed autolytic tissue. It can be a jarring sight.

I think perhaps you did do a rough investigation and for some social /ego interfaced reason you wont be forthcoming.

I sure hope im wrong as Im sick of this bs.
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  #44 (permalink)  
Old 09-25-2019, 08:04 PM
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Default Re: Strange U shaped lump

We have forums like this to post Observed Data.

Otherwise its only conversational clatter.

Principles; not "Personalities".
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  #45 (permalink)  
Old 09-25-2019, 09:18 PM
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Quote:
Originally Posted by Kmc View Post
So whats going on with this?

I dont buy the need to mail/do/ histology/freezing bs. This is what I find disappointing about internet forums.

Here we had a clear opportunity to at least LOOK at something. Everyone knows that freezing destroys tissue character.

I think you found Bones with helminths found or otherwise visual through collapsed autolytic tissue. It can be a jarring sight.

I think perhaps you did do a rough investigation and for some social /ego interfaced reason you wont be forthcoming.

I sure hope im wrong as Im sick of this bs.
Ive been more than transparent with this situation so for you to assume im not being forthcoming is beyond strange and offensive. Those assumptions are far out of line and really seem to come from nowhere. I personally dont care what "you buy", or "what you think x2". You dont know me. You only know what i have put out on this thread. Stick to the text, stop trying to imply things about other people.
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Old 09-25-2019, 09:37 PM
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Default Re: Strange U shaped lump

I am going on only what you have posted - including the actual title of your querie " Strange U Shaped Lump" Anyone with the slenderest bent toward biological curiosity would have simply peeled the dermis back - I did such things at the age of 12. So did so many others here.

Especially when you have stated how comfortable you are with dissecting. Peeling a little swath of skin back wouldnt interfere at all with your histological "ponderings"
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  #47 (permalink)  
Old 09-25-2019, 09:38 PM
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Default Re: Strange U shaped lump

I communicate on forums as I would in real life. Sorry for the inconvenience.
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Old 09-25-2019, 09:49 PM
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You also made no mention of histology until I mentioned its probable irrelevance in this format. No mention at all.

Then you went on and on about it wasting time.
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Old 09-25-2019, 10:04 PM
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Default Re: Strange U shaped lump

Are you sure you are going off of the thread? Originally, i wanted the vet to do a necropsy, so no, i was not planning on doing any dissection myself. After realizing this was not an option, i then decided to do it myself. Seeing as how i did not have tools or a scope on hand, i had to go and purchase one myself, adding a couple more days so i decided to move the frog to the freezer which i assumed would keep everything intact for later, i was wrong. Whoops. After learning that piece i turned to how i can preserve it for observation after being removed from the freezer. The formalin took some days to arrive, and then on the day it arrived i learned i was not qualified to carry that part out without proper equipment/material. This was Yesterday i believe. I discussed taking various samples before you added, "I dont think a complete histology series is expected." So you are wrong but what is your point? I thought it might actually bring some answers until i learned later i ruined those chances by freezing.

From what i am getting, you have it in your mind that A.) I anticipated my frog dying, B.) was prepared to dissect the frog immediately after death and C.) knew all of this information prior to my frogs death. This has been a learning process as i figured it would be and continues to be. I am not obligated to do any of this nor do i enjoy the idea of having to open up a beloved animal. Every reason for a delay is documented in this thread so again i dont know how you would get that im making stuff up to stall and make myself appear better? If that were the case i probably would not have even began this thread.

Last edited by PhylloBro; 09-25-2019 at 10:25 PM.
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Old 09-25-2019, 10:07 PM
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Default Re: Strange U shaped lump

What point are you trying to make right now? Are you trying to expose me or something? lol
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  #51 (permalink)  
Old 09-25-2019, 10:30 PM
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No, i anticipated your frog dying.

Hey the internet it is what it is. It could be different but human nature is a pungent, sticky master.
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  #52 (permalink)  
Old 09-26-2019, 04:32 PM
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I did it. It was bone like i thought. When i opened it up the bump that was poking the skin was the joint from its arm sticking up. I will post the pictures when i get a chance.
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  #53 (permalink)  
Old 09-26-2019, 06:31 PM
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Default Re: Strange U shaped lump

I was pointed to this thread by a DB member. This is the first time that I have logged in to this site in 4.5 years, and unfortunately due to time constraints I will not be able to make this a regular habit. But as this is something that I know quite a bit about, I figured that I would correct some misinformation. For anyone that I am commenting on, please take no offense to any statements. I just want to make sure that there is an accurate statement response to some of these statements.

Quote:
Originally Posted by ghartman View Post
Not sure what you decided to do, but if you still have the body I would just go ahead and cut man. if its been frozen, histology is not going to be very helpful. Most bodies that are necropsied in vet med are done immediately post mortem or within 24hrs (refrigerated) if you are looking for histopathologic cause of death. If it is a gross anatomical abnormality you should be able to identify it.
There is truth to some of this. Decomposition that begins to happen immediately after death rapidly limits what can be determined from a complete necropsy. This is particularly true for frogs, which autolyze extremely quickly. Essentially, the moment you find your frog dead - you need to immediately get it cold. The easiest way is to place it in a sealed plastic bag that water cannot get into, and then place that bag in a container of ice with enough water in it to cover the ice, bury the bag in the water/ice mix, and get it in the refrigerator.

Following that, you need to find a way to preserve the frog unless you are able to get it shipped overnight to a pathologist who can do a necropsy. Then the frog needs to be shipped in an insulated styrofoam container on several ice packs for next day delivery. If all if this is done, a full complement of tests can be run - bacterial culture, virus isolation, fungal culture, PCR, gross necropsy and histology.

However, in most instances - the best thing to do is to place the frog in a preservative (something to prevent further decomposition). There are two options for this:

1) 10% neutral buffered formalin. Pros - it fixes tissues quickly and has little artifact of fixation (only important for the pathologist). Cons - it is harder for a hobbyist to find, it is a toxic chemical that must be handled with caution, if the frog is left in formalin too long, it can affect future testing. Frogs fixed in formalin allow for a subset of tests to be run: Gross examination, histology, some PCR

2) 70% ethanol. Pros - fixes tissues quickly and allows for more postmortem testing (greater options for PCR testing), and it is easy to make [Dilute 7.5 parts grain alcohol (Everclear liquor) to 2.5 parts water). Cons - some tissue artifact, prevents some tests such as culture. Frogs fixed in formalin allow for a subset of tests to be run: Gross examination, histology, some PCR

Freezing is a last resort, as previously mentioned, it does introduce quite a bit of tissue artifact. However, it is better than nothing. Masses (such as the one in this frogs) can still be visualized and characterized, and infectious agents (such as bacteria, fungi) can still be seen. Viral infections are harder to identify if the lesions are mild. But, a spectrum of lesions may suggest a virus. Frozen animals allow for full PCR, gross necropsy, histology, and some culture opportunities.


Quote:
Originally Posted by Kmc View Post
A decent necropsy can be done with a pair of short, sharp scissors and some common tweezers. I dont think a complete histology series is expected.
Very few final diagnoses as to the cause of death can be made on a gross examination (cut it open and look). Amphibian tissues are so small, there is often little that can be evident to the eye. A dissecting scope will help, but it is still a poor way of doing a necropsy.

It is about as useful as your car breaking down, and you open the hood to look and see what is wrong. A small amount of time you may find the cause (busted radiator, blown water pump, etc), but you will almost always need a more in depth examination.


Quote:
Originally Posted by PhylloBro View Post
I just bought a kit and a microscope. I have a work around idea. I once reached out to an entemology department and was able to speak directly to a professor. If i can collect samples and take clear images, maybe i can send these or share these on multiple platforms and get intelligent responses from qualified people.
This will not be very useful. Tissues prepared to look at under a microscope need to be preserved, processed, thin cut and stained and then examined by a professional. From just looking at the outside of something, someone may be able to tell you that there could be something wrong - but not what is causing it (unless you are lucky enough to find a worm right in the middle of it - which is unlikely in an amphibian - and not likely in your frog).

Quote:
Originally Posted by ghartman View Post
If it is fungal, bacterial, etc. then fixating in formalin will not help you need fresh tissue.
This is not necessarily true. Between a standard necropsy and histology with special stains to highlight certain characteristics, along with the pattern of lesions, much can be learned. If more info is needed, PCR is possible in many cases, that as long as there isn't excessive time in formalin (more than a few days) will still work quite well. You are correct in that you need fresh tissue for culture, but that isn't always necessary.

Quote:
Originally Posted by PhylloBro View Post
Called the vet and asked if they do necropsies. They said they no. They only do them for birds and they arent in house. They send them off to a university to have them performed. She said they could do tissue samples but it would cost hundreds or thousands of dollars. Im assuming thats what she meant but she literally said hundreds of thousands of dollars lmao Ill search around but if anyone knows of an institution who performs necropsies on amphibians let me know.
Necropsies can be quite expensive on large animals (at some places, up to a couple of thousand dollars). This is not true for all animals.

Amphibian necropsies are available through the Aquatic, Amphibian, and Reptile Pathology Program at the University of Florida. A frog necropsy of any type of dendrobatid is just under $70. This includes a complete gross necropsy and full histologic interpretation of all tissues and any special stains (done to identify fungi and bacteria). For larger frogs and small reptiles, you are looking more at $80-100. Only the largest of reptiles (or an adult giant chinese salamander if you have one - and if that is the case I will do it for free ) would be in the $200-300 range (adult retics, burms, old sulcatas, aldabras, and galaps).

I am a veterinary pathologist who specializes in amphibian and reptile infectious diseases. I started on Dendroboard in the early days of the forum as a Vet Student. I have a strong passion for the care and health of amphibians and reptiles which is why I do what I do. I have worked at the Bronx Zoo, Shedd Aquarium, Brookfield Zoo, and Lincoln Park Zoo and now receive cases from around the country. I helped start a program at the University of Florida so that hobbyists would have a way to get help when they have sick animals. For most institutions, necropsies need to be submitted by a veterinarian. However, if you are willing to pay for the necropsy up front (to the University of Florida, I do not make money directly from any necropsy I do), and you will fill out the appropriate paperwork and package the animal properly - I am happy to help. I have already done this for numerous people in the hobby.

To find out more about our program and my background - you can click here https://labs.vetmed.ufl.edu/services/aqarpath/
and here
https://www.vetmed.ufl.edu/about-the...bert-ossiboff/

If you would like to submit an animal for necropsy, I can be reached at the email address in my bio. I also work with the ZooMedicine Diagnostic laboratory where we have the ability to test for Bd, Bsal, ranavirus, and other parasites.

I do not give out treatment info or drug information based on pictures or descriptions. I am sorry - but it is illegal to do so, and inappropriate medical care. I also am not likely to respond to PMs, so email is the best way to get in touch with me.


Robert Ossiboff, DVM, PhD, DACVP
Assistant Professor
Aquatic, Amphibian, and Reptile Pathology | ZooMed Diagnostic Laboratory
Department of Comparative Diagnostic and Population Medicine
College of Veterinary Medicine
University of Florida
Gainesville, FL

Last edited by rozdaboff; 09-26-2019 at 06:42 PM.
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  #54 (permalink)  
Old 09-26-2019, 07:15 PM
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Can't thank you enough for stopping by, Oz.
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  #55 (permalink)  
Old 09-26-2019, 07:44 PM
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That post is the most useful thing to come from this thread. Thank you
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Last edited by PhylloBro; 09-26-2019 at 07:46 PM.
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Old 09-26-2019, 08:53 PM
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In my boots on the ground experiences, since 1981 having been teched and trained in the old fashioned way by the reptile veterinarian of the reptile collection Bronx Zoo, then later at the Ca Oakland Zoo, and my own conservative explorations. I do understand that a rough visual is not ideal but if visual parasites were visually apparent, it would be better than no visual investigation at all.

I will go out on a limb here and disclose that I have done as personal quest per rough necropsies, and have determined mechanical sub ingest, parasite, and even some other causes. (Polymer crystal ingestion, journaled)

I will go out on a limb even more to say that over exuberant supplementation causes harm, and is in vogue applied per presentation of worrisome mysteries of health matters. Causing toxicity and death - Rarely investigated.

Last edited by Kmc; 09-26-2019 at 08:55 PM.
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Old 10-10-2019, 11:04 PM
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Sorry for the huge delay I’ve been super busy with life stuff. Here are some pictures I took of Bones. As you can see, the light grey is the bone.
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Old 10-10-2019, 11:08 PM
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Here is something interesting I found. Near the bone injury was this strange looking growth. It was like a bubble that wouldn’t pop. I could move it around and it would keep its shape. It’s possible I knicked something and didn’t notice but as far as I know it was present as soon as the bone was revealed. Here are pictures
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Old 10-10-2019, 11:11 PM
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To prove that what the “U” was was actually bone, I removed it completely. Here is the scapular. I also removed a loose bone shard that was near the injury. I determined it was bone by feel so yeah lol but I’m positive it’s bone.
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Old 10-10-2019, 11:14 PM
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I just noticed the image quality is terrible. When I get a chance I’ll try a different method
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