# Sticky  Instructions for preparing, and source for, necropsy



## rozdaboff

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.



ghartman said:


> 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.




Kmc said:


> 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.




PhylloBro said:


> 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).



ghartman said:


> 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. 



PhylloBro said:


> 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-college/faculty-directory/robert-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


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