Episode 201: A Backstage Pass to Freezing Semen + Postmortem Sperm Rescue

Andrea Hesser, DVM DACT gives a step-by-step walkthrough of a frozen semen appointment to help you understand what to expect. In addition, learn what can be done to save sperm postmortem or in crisis situations.

Watch the video version of this presentation here.

Transcript

Nicole Engelman  00:08

Welcome to the Good Dog Pod. Join us every other Wednesday when we discuss all things dogs, from health and veterinary care to training and behavior science, as well as the ins and outs of Good Dog and how our platform can help you successfully run your breeding program. Follow us and join Good Dog’s mission to build a better world for our dogs and the people who love them. 

So, hi everyone. I am Nicole. I'm Good Dogs community lead for those of you who don't know me, and today, we have a really exciting presentation with Dr. Andrea Hesser, who has graced us with her presence many times throughout the year, and has hosted some incredible webinars for us. Today's is no exception. It is a backstage pass to freezing semen and post mortem sperm rescue. So quick overview of what that means and what we will be covering today: Dr. Hesser is going to give us a step by step walkthrough of a frozen semen appointment to help you understand what to expect, as well as share what can be done to save sperm post mortem or in crisis situations. And we did get a ton of questions related to this topic during our Q&A roundtable at the end of our symposium, so it felt very timely to have a whole webinar dedicated to this topic. So please continue sharing lots of questions throughout the presentation, because it really helps us have a great Q&A that everyone can benefit from. I want to thank Purina, of course, for helping us bring this webinar to life. As I mentioned, this really was brought to life because so many of you have asked questions about it, so please continue asking questions, sending us emails with topic suggestions, because it's so helpful for us. And crazily enough, we are planning for 2026 and we have a wide open calendar right now of content and ideas, so you can help shape what we cover next year. All right, I will just quickly mention a little bit about Good Dog for anyone who is new here and joining us for the first time: Good Dog is on a mission to build a better world for our dogs and the people who love them by advocating for dog breeders like yourselves, educating the public and promoting canine health through events just like this. We are a community built just for responsible dog breeders to connect with quality Good Dog applicants from all across the country and find forever homes for your puppies. We help breeders run all aspects of their programs from start to finish. We have tons of past webinars that we've recorded in our Good Breeder Center that we can drop a link into the chat now so you can look at those. We have educational articles, a podcast. We also have health related discounts as well, just to help your program thrive in other ways besides placement management as well. So if you're not yet a member of our community, we would love to have you join us, and we invite you to learn more about our mission and apply at GoodDog.com/join or just reach out to us at breederteam@GoodDog.com if you have any questions. All right, before I pass things over to Dr. Hesser, who I hope many of you are familiar with already, I do want to share a little bit about her background in canine health. Dr. Hesser is a board certified reproductive veterinarian who practices in small animal medicine in the Dallas–Fort Worth area. Her clinical time is spent doing general practice veterinary medicine and reproductive consultations and surgery. Dr. Hesser also provides semen collection and health testing services at local dog shows. She's a longtime canine breeder and enthusiast with an interest in Whippets and Bull Mastiffs. Her mission is to serve breeders by helping them achieve their goals with their breeding program. So nobody more qualified to be here today to speak on this topic, especially; so with that, Dr. Hesser, I will pass things over to you for a presentation.

Dr. Andrea Hesser, DVM, DACT  03:45

Thank you, Nicole, and welcome everybody to the seminar. I am going to spend most of my time just so you know, at the start, we're going to go through kind of the process of freezing semen from start to end. A little bit of that's going to be review of some things that you guys already likely know about semen evaluation, which is going to obviously be important before we go forward to freeze semen. And then towards the end of things, we're going to discuss and go over some procedures that can be done after either neuter or post mortem, potentially, so that procedure that can potentially rescue and then go on to freeze that sperm, should you ever run into that need. So as she was saying, we're going to go over some odds and ends relating to the freeze process, a backstage pass. Mostly the focus of this is to show you the things that happen when you're not there. Obviously, when you're there for your appointment, you're there for the evaluation, oftentimes for evaluation for other things too, right? So that stuff, you guys probably have a lot more familiarity with. I'm going to show you what we do kind of after that time and how the process goes. We will talk a little bit about some of the common things people ask about: different types of extenders, pellets versus straws, how they differ and how they look, as well as some just discussion about storage and shipment. That's going to be more on a logistical note, but it's obviously very important for you guys. And then a little bit about use. This could be an entire lecture on timing for frozen semen for an hour, but I just wanted to mention the highlights of how we would go about using the semen as well. If I had to pick a topic that is my baby, this is probably it. I have done, probably–not to toot my horn—but I probably in the United States, I've probably done more publishing on cryopreservation type papers than anybody else from my residency. So my residency, I actually did as part of getting my board certification, I was very involved in collecting and freezing dogs from a local facility that we had access to that has guide dogs, so Labrador colony, essentially. And we did lots of odds and ends and testing. I've also been involved with a couple different breed clubs that have hired the Davis team or me to come in and do freezes or seminars or studies at shows, their national shows and things like that; Great Dane club being the most recent one that Dr. Myers had worked with before he kind of started to retire out of his Andrology lab. And then certainly case reports and things I did present, there's not really any data on use of the epididymal sperm, and I don't have that paper here, but I did present on a case where I actually did the kind of pre-mortem, but I had a two-year-old dog that had a neck injury in the field. We did a firm rescue procedure on him before euthanizing him. Then we went to use that semen later, and then produced a litter from that. So I've kind of been on both sides of that picture, which is kind of nice, as far as review. 

So as far as why we might be freezing semen—I know this is really basic. Obviously, you guys probably have frozen semen with your veterinarian, locally. It could be that you're already very aware of these types of things. But a lot of people, the reason that people come to us to freeze, is oftentimes when the dogs have already had accomplishments, they've already had health testing, they've already produced litters and made a name for themselves as a stud dog. Sometimes when your dog has gone from being, you know, a two-year-old dog that just finished his championship, and now he's a six-year-old dog who's been out with a handler for three years, and he was the number one Chesapeake Bay Retriever in the country for that entire time—well, that dog is going to be a six year old dog now, right? He might be a more accomplished stud dog. He might not. But he might have older signs of his sperm count, his morphology might not be as good as it used to be, his prostate might not be as happy as he was when he was an 18-month-old dog. So I think you know, for poking holes and saying there are some mistakes made, we see lots of people who come to us and say, I have a 10-year-old dog and I've never frozen semen on him, and have great expectations of what they'd like to freeze when what they've done is waited until the dog had accomplishments, which is reasonable, but you guys are super polished experts in your own breeds. You know, when you have a really good dog that maybe is clear for everything, maybe his health testing isn't complete yet, but you anticipate, based on his lineage, that he's going to be in good shape. So I think my message to you is think ahead, because we see so many older dogs that we might be freezing, like I've got a Wire Fox Terrier right now. He wants to freeze something like 10 breedings, and we're getting maybe 0.75 or so of a breeding each time we freeze him. And that's per the US standard. She wants to ship a bunch of semen overseas, so that's going to take a lot of effort. Meanwhile, she's got a three-year-old dog that she's freezing at the same time, whenever we're seeing the other dog. Well, the three-year-old dog is producing more sperm that's healthier, and he's freezing at higher post dots. So, you know, sometimes we have real life lessons in front of us, but it is a good path to have testing ahead of time. I'm not trying to say like, freeze your one-and-a-half-year-old dog and don't do health testing, but I think sometimes the semen was better when they were a little bit younger. 

So, again, that young dog that you're thinking about starting to evaluate, maybe he doesn't have all the ribbons in the show career, but you know he's going to be a good dog, or, you know you're about to send him out to accomplish those things. Freeze him before he goes. He'll be more available for people once he's showing if his show schedule conflicts with his ability to serve a dog that would normally do a natural breeding, maybe is on the other side of the country and sees him showingone weekend, and then he's traveling to another area of the country, and isn't going to be available . . . A lot of people that are either hunting their dogs on a high level—you know, if they're going to a local trial in certain seasons, those dogs might not be available because the trainer says, “No, we're not doing any natural breedings whenever he's working.” And that's something you have to work around. 

I think the other major thing that we use this for is international work. So, say, I have a Bull Mastiff that you guys have met. Cabana. Actually, one of the boyfriends that I considered for her, her great grandfather is a dog from Denmark who has frozen semen in Denmark. So I considered shipping semen from Denmark for her, like a while back, I didn't end up electing for that because it didn't end up being my preference of the dog once it was more mature. But I was thinking about using that line breed, and would have been a really good match for her on paper, based on their background, she needed to be line bred, kind of with where she was. It's a lot of expense, certainly to import semen from Denmark, but it's nice for that particular breeding that I had in mind. It was a really good kind of thought for her. So otherwise, you know what people used to do: people might ship dogs overseas, or ship a dog to show for a little while, or to trial for a little while, breed the dog while it's over there, and then, before it actually was pregnant enough to not be able to get on the airplane that they'd ship the dog back pregnant. And sometimes that's, you know, sometimes we're doing that nowadays, but that used to be the only way that we could work this. And now it's a steep bill. Don't go into it thinking you're not going to spend a lot of money. I would say, the air bill to ship something overseas, usually one way with the tank rental and the FedEx bill, I would say, is usually $1,500 to $2,000 so I mean, you're looking at a chunk, but a lot of companies can help facilitate that. Sometimes you can get kind of group shipments. It's not always on your shoulders as one person. The other thing I would consider for is things like, if you freeze and keep your own stud dogs, we have some clients that will say they have a two-year-old dog that they like. He's not their favorite. They have his brother, and they like his brother better. They might freeze the brother, or they might freeze the one that we're talking about. They might freeze both of those boys, even though they're going to place one of them at a companion home and he's going to go be neutered, you know, in a few months, to go be a pet for somebody. Well, maybe retrospectively, he was better genetic match for something years from now that you go and want to do and want to produce. I think sometimes, you know, it might be that one dog lived to see a 12-year-old lifespan in a giant breed as a male, and the other dog got cancer when he was five. Even though we liked the dog that got cancer when he was five better structurally, if they were similar on paper, you know, I'd be choosing the frozen from the dog that had longevity. So I think it flexes our ability. The longevity thing, I think, is a huge one that is underestimated. But I think that might be the golden ticket to get some of these breeds out of their longevity crisis holes that they're in. I mean, Bull Mastiff is a beautiful example. We have plenty of dogs that live to 10–12 years old, but our national—I can tell you, when we go there might be a dog in the 9+ category. That's our oldest category. We don't have, like, for Whippets, it’s a 9–11, I think, and then 11+, maybe 11 to 13, then 13+, like, we've got a really high age categories, because we actually do have dogs that come for those shows with those ages. Bull Mastiffs, we are lucky to see, especially on the male side—we're lucky to see a 9+ in the show ring that's presented, and maybe that's because of their condition. But I think for the most part, our life expectancy is decreasing over time, and we're seeing more and more docs with cancer. Well, it could be popular sire syndrome. It could be certain lines. Maybe it would be really valuable to have some of these lines that we didn't think we would need, or that we didn't think that we would want.

13:39

All right, so setting up and scheduling for your frozen semen appointments: for the ideal candidate, younger to middle aged dogs. We do freeze older dogs. Not to say that we can't freeze older dogs. I get surprised all the time at how they perform. I just will always have a conversation. If I have a dog that's more than five or six years old, I will say sometimes, you know, we expect their post-thaws—I'm going to project for you their post-thaws are, you know, going to be 50–60% or better for the average young dog with typical breeds that freeze well. We're not going to see a 75% post-thaw in a 10-year-old dog. It's just not going to happen. We might see one in a two-year-old dog, but by the time he's 10 years old, he might post-thaw at 20. Some dogs start from the get go. They don't post well, but I think the older they get, the more fragile the semen is for processing. It's just the way it goes. Ideally, high sperm numbers. So you guys that have toy breeds, if you have toy breeds, we are looking, oftentimes, we are looking at partial doses of frozen semen that we're using each time. So we might be freezing a dog that has 75 million total sperm before we freeze it, not including the factoring in the motility, so doing all the math, that would make that even less of a dose. If we say we want 100 million modal sperm for a breeding, obviously, that's going to take some work to do that. Meanwhile, the other end of the spectrum, like I had a Golden Retriever a couple weeks ago that collected 1.8 billion sperm. That is way high for Golden. We don't collect Goldens that produce that much sperm. Generally, that's the most I think, in that breed that I've ever collected. And I've collected a lot of Goldens over the years. But you know, we might see an ejaculate like that in a Great Dane—the really young, healthy Great Dane, or a Mastiff, or something like that. It's usually associated with body size. You know, their testicular size is correlated with their sperm numbers, generally, in healthy dogs. So for that Golden Retriever, well, he ended up getting six or seven breedings, which is an outstanding result if you're really doing proper packaging and using ferment appropriate doses with accurate post-thaws, it is very difficult to have a dog that produces that many breeding per collection, despite the numbers that people are often quoted from certain companies that tend to make this a very difficult conversation. But other things, so I mean simple things: We love it if they have great motility, they freeze better. If they start high, they're going to end higher, usually. And morphology, we tend to factor significantly. So if they're not at least 70% modal and 70% morphologically normal, it's not a fool’s errand. And it doesn't mean that you can't freeze them. I would just start asking things to the clients like, “Do you have any recent litter from him? or “Is he actively being used and producing?” If he is, then I would feel comfortable, you know, from that perspective, saying this is going to work. You know, if you go to use it, and you use enough of it, it's still going to be able to work. Some of them, we have to kind of freeze on a “we hope this works” basis, because some of these dogs, you know, if they're not being used in the proximity of the freeze, and their morphology doesn't look great—I mean, it might be that their fresh wouldn't have worked. So we can't predict that for sure. All we can say is, you know, if they're close to normal, then we're happier, you know, starting out. And then, I mean, a silly thing to say, but no concurrent health issues. Fine if they're on medication sometimes. I think we get a lot of questions like, “Oh, do we just had a UTI and he's on Baytril right now?” Well, that's fine. It's not going to hurt the sperm to be on Baytril. But did the UTI cause a prostatitis, and he's got prostatitis on top of it? I had a Labrador that was a field trial dog that had a chronic skin issue on his side, and eventually they figured out it was a foxtail, but it was like an abscess. They drained it, they flushed it, they tried to find something, couldn't find anything. Eventually, we removed it like a mass. But during that issue, the dog had poor morphology, and even though he was managing actively, we just had a lot of caution with “is this going to be good sperm?” We don't know. Fortunately, he was being used kind of concurrently during that process, so we knew that it was probably okay to be freezing him during that.

17:54

This is a note; I think this is kind of a given, but usually most dogs—not all dogs, but most dogs—really like to have a teaser present. They're very excited about it. It doesn't even have to be a teaser that is in season. It doesn't have to be a female dog. Some stud dogs really just like being with their housemate, and that makes them happy at the clinic. So they like to collect when they have the housemate around. Some dogs that really understand bitches in season have to have a perfect, perfect dog that is exactly what they would breed naturally. They have to mount her, and they have to think that they bred her in order for me to get semen out. I mean, sometimes that's just the way it goes, but I would say most of us prefer to have a teaser bitch present. Some dogs need a lot of contact. We got to make sure those dogs are both brucella negative for us to be able to do that. You know, it comes from the owner's house and it's already exposed to that dog. Contagious diseases can transmit from pretty subtle contact—brucellosis being among them. We don't want that stud dog to be licking all over her vulva if we don't have a negative test on both of those dogs. He could transmit it to her. She could transmit it to him just from that contact. So just consider that there's a lot of repro clinics that do a lot of volume that will shove a bitch that's dropped off for a progesterone test right in the face of a stud dog and not ask any owners any questions. I do not think that that is good as an infectious disease approach. It is a terrifying idea. So you guys are attending a webinar, right? You care. You're involved. We see a lot of breeders that, I mean, they're not the good kind, right? Like, we don't see 100% of the good kind. There's a lot of people who don't do brucella tests at all, that will breed their dogs that have active health concerns and not tell us. They might be breeding a dog that has kennel cough and does not want to say it, because they want to breed that dog this cycle, and they don't care, and they didn't think it was an important thing to mention. I mean, there's just stuff that we don't want to birth from one dog to another. So not to mention sometimes some dogs react aggressively toward each other if they're nervous or scared or reactive, because that's how they are. So all the more reason; just be aware of that. Be careful. Ask those questions of your veterinarian, of anybody that's collecting your dogs. Make sure that you're in control and that you're present, because things like that happen. The thing we want is to be able to collect your dog successfully. So I think some clinics that get busy, just forget to be more cautious about that. And they'd rather have a happy outcome for you, even if the detail got dropped. 

So before your appointment, this is going to be things that you're going to want to do, pretty much regardless of what kind of dogs you have, if they're AKC, for the most part: Registration numbers, no matter what kind of club you belong to. If you're AKC, if you have a breed association, if it's something where—like, every once in a while, we'll have a club, I think it was American Pit Bull Terrier club that is not AKC affiliate, obviously. Those guys have some kind of a specific document that we're supposed to sign. Like, just make sure that if you know that you have something special that needs to be done, just make sure that we know that so that we can help execute that for you. The microchip number is mostly important for shipping semen overseas. So some countries will require the microchip to be on the straw, or they will literally throw it away when it reaches the border. There are certain requirements that are very specific for some of these types of things. We don't routinely put the chip number on the straws, but we do include it in the contract paperwork, usually. AKC DNA profile, so that is going to be AKC specific, obviously. If you're planning on registering your litters with AKC, they require any frozen semen use. They require a DNA profile to be on file that is not your Embark profile, or your Paw Print profile, or your Orivet profile, that is your AKC parentage DNA. I think it costs $50. You can pre-order the kits off of the AKC website, super easy to do with the peak swab, and the owner can do it at home. We used to supply them, and then they started making us prepay for them, so we no longer purchase them ahead of time for clients. So we just remind and remind, but just make sure that you obtain that. You don't have to have it at the time of the freeze, right? So it can be something that you can do later. Don't forget, because if they go to register your litter and you never got one, never had one on file, they're going to say you have to thaw some semen to use for that. They can do that, but you're going to waste some semen that you spent a lot of money and froze with purpose to use for breeding, not for DNA. Whenever you could get a freebie sample from your live dog, it's kind of a crummy position to be in. It is possible to do, and super easy if you know where to send and all that sort of thing, for the vet. Front and side images of the dog, it's good practice. We don't really do it, to be fair, anymore. I think with chip numbers, and you know, how the world has changed in 30 years, the West has been doing it that way for a long, long time, so they implemented that back before everybody kind of had a microchip or had more organization with their clinic. The reality is, is you could bring me a dog, any dog under whatever registration form, like you could show up with your dog, and you could bring his brother, and I could scan him, and if I didn't have a microchip number on his registration form associated with his other paperwork, how would I know that it's two different dogs? And we could take pictures of the dog, could still have been subbed out and been a different dog, right? So that's kind of the thought, why they do the profile on all these guys. And then we require for anybody that we freeze, I think it's good practice to be negative for brucellosis within six months of the appointment. I think if you're doing a lot of work, like breeding your dog frequently, I think having one more regularly might be a good idea. No test is going to be 100% accurate. So even if I say, you know, he's Brucella negative today, he could have been exposed to Brucella three weeks ago, been sick, gotten better. Look fine on his semen. Some of them can, and then we can freeze that, and we can freeze bacteria and bring them back to life, like the sperm cell. So just be as judicious as you would. Think of this as just like you're bringing a stud dog for natural cover. If you feel like it would need to be updated, we'd recommend updating it.

All right, so we'll kind of breeze through this, the first phase of the thing. So this is kind of into the process itself. So we're obviously going to collect semen, then freeze the semen. Any collection method is fine. So everybody has their own preferences. I like the plastic sleeves that are disposable because it's just easy, and they're inexpensive and easy to acquire, easy to stock. The cones on the right. Some people like those. You have to clean them between, obviously, if you use them. I also worry about trauma to the tip of the penis, depending on how coordinated you are. Some of these dogs that they're thrusting really actively, it's really hard to keep them from kind of knocking into some of these edges on these cones, and they can get focal bleeding from that if you're not really used to them. I would definitely practice on a non urgent, non freeze, non fill appointment. If you guys get these cones, like, if you're a veterinarian listing and you're interested, or if you're an owner that's interested in using them, the nice thing is, they fasten right on to these little 15 mil conical tubes that we use for collection. And you can switch between, like, if you had urine in the sample, you can switch it out and, like, have a new one ready, and be able to section that off a little bit faster than maybe with a sleeve. But I like the sleeves. The left side, that's just a latex rubber. It's called a latex artificial vagina, which just basically is the term that we use on the equine side. It is made for this purpose. Basically the penis will swallow inside the sleeve. And then you can take the tube at the bottom. You can pop the tube off and leave the sleeve on there, so he's got protection while he's still erect and he's outside of his pants. So it's kind of nice from that perspective. Downside with those, they have to be clean. You have to have a whole bunch of them. So you use one, and then your tech has to handle the cleaning part of that. The cleaning part is not too bad, but if there's any soap residue, soap will kill sperm. So that's bad, obviously, for the next dog. Also the latex itself. There are some dogs that their sperm is allergic to latex, and they will die like soon after you collect them, the sperm will look dead just because you collected them in latex. I'm sure they could do that with plastic too. I've just never seen one. Just FYI, there are other ways to collect. So there are techniques with a catheter under anesthesia that are possible. So like for a dog that's in a critical condition that cannot ejaculate on its own but is still stable, it is possible to do that. Probably I'd say most of these guys, they have something like that. We end up doing epididymal flushes, which is what we're going to talk about at the end. It is possible to do something called electro ejaculation, which is primarily a technique that we use for livestock. So we actually use EEJ for cattle that are getting semen evaluated, for example. When they're in the shoot, they'll insert something rectally that causes a stimulation, and then that causes them to have an ejaculatory phase. I have had some experience with this the EEJ and dogs. It requires a very specific probe and some very expensive equipment. So these units are, I think it's something like with the probe and the unit. I mean, this is like 10 years ago too. Probably like 20 grand to buy this. So usually it's at a teaching hospital, because they're using them for, like, several species, different departments that might be able to use them. And obviously, because it's kind of a tertiary procedure. The other thing that we can use to get collections is Lutalyce. It's a drug that we use injectably that can improve libido temporarily. Some dogs respond really nicely to that that are difficult to collect otherwise. 

So dogs will collect in three different fractions. So we really only care about the sperm rich part. So the first fraction on this example, so you can see, there's like a little tiny bit of fluid on that first fraction on the left; the middle tube has the really concentrated sample. And then the third fraction, you can see above the like green top, you can see where the flute line is. So we collected some prostatic fluid on that particular dog. We're really mostly interested in the second fraction, obviously, for the freeze part, the third fraction we might be interested in if we thought he had prostatitis. 

So we're going to evaluate for motility. I know it's kind of a given. So it just depends on your vet, whether they use something computerized, whether they use something where they have these, like marking systems, where the computer will, like, pick up the dead cells versus the live cell. These are not perfect, and you do have to adjust them. You have to calibrate them, you have to know what you're looking at. But like for this one, for example, there are some cells that it’s double counting. There's one cell that has two red dots on it. It's counting that as a dead cell twice. It's also counting one of the cells that's moving kind of on the right side of the screen, there's a couple. They're not moving fast enough for this computer to decide that they're even alive, which is not correct. Like these systems, even though everybody loves these graphs and these like charts that they produce, and a lot of practitioners start just saying, like, “Oh, this is a tool that we can basically remove ourselves from having to be involved with this.” It basically just creates more work for us. I don't use them at all. Totally honest. I thought about getting one just to let clients see the picture, because the thing is, like, the microscopes are wonderful quality that are associated with these. They give you beautiful images like this, and they have, you know, ways that you can save images and save videos. The problem I have is, if I use this technology, I will end up adjusting it to what I think that motility is, because I think my eye is sometimes going to be able to be more sensitive than what the camera is going to pick up. Well, that kind of defeats the whole purpose, right? And if you accidentally read it wrong, if I turned a certain parameter up or down on this one and made this say it was 99% modal, which is what I think a lot of what happens at some of the trucks that drive around at the shows have all these reports that are wickedly high, because I think the concentration is super high. The computer can't even read the cells, and everything looks like it's moving, because if one cell bumps into another cell, everything's modal. So that's the problem with these units. But if I record that, say, under your dog, Blue—If Blue had a 75% motility last time, and I record him as having a 99% I can't get rid of that in a lot of those systems. So it will look like, graphs over time, it'll look like suddenly he was so much better this appointment, when really he wasn't. 

But other things we're looking at: sperm count. There's lots of different ways they might look at sperm count. One would be using that CASA technology, so that imaging technology, again, look at the holes that you could poke through about just that one view that we were looking at. It was double counting some, it wasn't counting others. Sometimes it'll count white blood cells or red blood cells as sperm cells. It is a problem with many of these technologies. Same problem we see with the photometer. So the photometer, that's the thermo cube, that unit is actually what I use. It does not work well if you are collecting semen that has blood in it or has white blood cells in it. So the only things that can work the best with those scenarios are going to be nuclear counter, which is super expensive, again, a five figure machine. All it does is count sperm. It's usually just university level that people have it. Or hemocytometer, which is when we sit on a microscope and individually count sperm cells on these little special chambered flies. Very laborious, very annoying. You have to have kind of the right setup to do it, but that's what I'll lean on if I have an abnormal sample. And in this scenario, if I have an abnormal sample, I'm not going to be freezing it, right? So like that, thermo cube works pretty well for my needs in a normal ejaculate setting. If he didn't bleed into the sample, if he didn't have any problems with his collection, then it's perfectly reasonable. If it's not, if it doesn't look good, and we're not going to be freezing it—so I usually use that and then just estimate down. If I'm not trying to be super precise with my numbers in that way, because I know that there's a little bit of red blood flow content, so I know it's going to over count a little bit on the concentration. 

And then the last thing we're going to look at, before we say, “Yep, he looks good to go,” is looking at these morphology images. So this is just an example of one that I pulled off the web, but essentially this is canine sperm cells. We're looking at the way the heads are shaped. Do they have the parts that they're supposed to have? Do they have anything extra that they're not supposed to have? In this picture, you can see what's called a distal droplet right in the middle. So there's a swelling in the backside of the midpiece on that cell. It might not be a significant defect to prevent fertilization, but we like to document these types of defects, even if it's a mild one like that. So if you go kind of straight across from that cell, and you're looking at like, three o'clock from that cell with the distal droplet, you can see a cell that it didn't stain very well, right? Like it kind of absorbed the stain that's in the background. It's not bright white. It's got some little vacuoles, so it's got like little circles in it. And you can actually kind of appreciate that the acrosome, which is the end part of the sperm head, like the very tip of the sperm head, that's supposed to react with the egg, almost looks like there's something floating above the cell in front of it, like a C-shape above the cell. That's a reacted acrosome. That cell is not going to do a thing. If we had super high motility, and you had 50% of those, that might be a big problem for that dog to perform well at all. So not going to be something where I'm going to say, “Oh yeah, you should spend a bunch of money freezing this dog today.” 

Just a caution on again, some of the less thorough clinics or ignition based trucks that are out there freezing: most of those clinics don't look at this. They will say they did a morphology of this, but they're either using that CASA system, which none of those CASA systems are effective at doing morphology evaluations, like, truly, when we look at it statistically, they're not representing, like, what we would rate it as when we do it by hand. With most of them, they're just saying, like, “Oh, I think It's like 10% coils.” But when they're looking at this, this is what they're saying, “Oh well, there's one bent back on itself, and then there's another one that looks like a couple of droplets.” That's what they'll estimate from. They don't do that so they don't see heads, they don't see detail. It's amazing that reports that we will get.

34:19

So we've evaluated it. Looked good. We're going to go forward. Next step is we are diluting that sperm rich fraction either raw, so they don't have to be centrifuged before. If they're super, super concentrated, we could go straight from that super concentrated state to diluting with the first step. I would say, the majority of the time, we need them to be pretty concentrated. So we will end up doing a centrifugation step at a low speed, kind of at a urine type speed, where we do the sediment speed, and then we draw the clear part that's kind of collected at the top. And then after we do that, then we add the first step of our freezing process. Most of these freeze extenders that are out there will have multiple steps, so they might have, like, a part one, part two, or have an initial buffer, and then, like, the one at the top is the lettuce one called the freeze buffer is the first one is at a certain ratio, and then the dilution buffer you do to a different ratio because they want to make it a certain concentration. The one right below that is the MOFA based one. It's the same now, it's a different name as the one that's in the bottom right corner. That's one I use pretty often. That's called Canopus. It is add A and B at the same ratio. It doesn't have a lot of rules that are stated by the company. I usually try to package it at a certain concentration. So I'm shooting for a certain dilution ratio so that I can read the sperm on the slide, and so that there's enough extender there to make them happy, because that's part of what we're doing. Some of it we're trying to dilute it to make it the right concentration to read it. But some of it is we're trying to protect it from dying through the freeze process. So egg is really common as a substrate for that. So these extenders, even though you can see some of them, they have opacity to them, so you can tell there's egg in it. The others are clear, right? So that is before we've added egg yolk. So we crack the egg yolk through a process of, like, kind of removing all the albumin on the outside of the egg. We insert a needle and draw the yolk directly out of the center of that yolk, and then we add that in certain ratios to these pre-made kind of extenders with the other component. So then they'll have the other components, like glycerin. So glycerin is a type of sugar that can be nutritious and helps decrease the crystal formation that happens when you freeze things. So glycerin is basically how we manage to get these guys to freeze and survive. And then if you have sugar and egg in something, if you guys have ever done any baking or if you are a science person, you know that egg and sugar are excellent media for bacteria to grow. We keep antibiotics in some of these extenders specifically for that reason. And just like a person could have a response to a particular antibiotic or an animal could, we don't tend to see a reaction to the antibiotic. I think I have one client who had convinced herself that the antibiotic free chill extender was the only way that her dog could ship but I don't think I've ever run into it. It's like with anything, some dogs will do better in one versus another. 

So other mentions on extenders. So one of the extenders that I listed here, so some of these are packaged. Uppsala is an example of one. We'll say, like, if somebody says, “I use Uppsala,” they're not buying an extender from the university. What they're doing is their university produced an extender recipe that is available publicly, and they have provided that online publicly. So basically, there are recipes like that that are out there. So just be aware that could be what somebody says. I love the Uppsala extender. We've had trouble getting ingredients for it. We have in the recent past. That's what I've used, you know, in my own clinics. It has been the Uppsala extender. I think it's a really excellent extender. But the downside is you have to have, like, micro scales, and it's very difficult to make. You have to acquire all these powdered antibiotics that we get from, like science, where you'd expect, like your science teacher to shop. That's basically the kind of thing that we have to do. 

aSo what is the best extender, right? That's the common question people will ask. So most of the commercial extenders will yield pretty similar results. I think the best extender conversation, I think Uppsala is probably on top when you can get the ingredients and when it's made properly. I would say the other one that I see incredible post-thaws on, that's not really around anymore, but we hear whispers about it, and we'll get older semen that’s from it. But there used to be an extender called Clone, which some people have suggested is basically the same as Uppsala. It's not exactly the same, because it does have an activator, so when you thaw it, you have to thaw it with the thaw media in order to wake it up. Uppsala doesn't do that. So I'm not sure exactly what the difference is between those two, but just FYI, those are probably the ones that I think have the best, consistent post-thaws. The downside with Uppsala, Uppsala does not like to wake up, so it doesn't like to look good initially. And it is common that we will have an owner if we ship semen that is Uppsala, or if we receive it, it will look dead the first moments that you've thawed it. And most other extenders look live right away, but it takes like five minutes in a water bath, and then it'll be like 80% post-thaw sometimes, when it looked dead when you first thawed it. Meanwhile, the opposite is true—well, not the opposite—but most of these extenders, like the Canopus, it looks good immediately, like immediately. We thaw it, and suddenly, it's magic. So I think that is a little bit of the difference of things on some of these guys. There are certain companies that claim all these super high post-thaws. Well, they're not getting those post-thaws. The paperwork says that's what the post-thaws are, but like, most of their dogs will freeze between 30 and 40% I would say, on their post-thaws, when we're thawing for use. So you can do what's called a freeze trial, a little bit of a pain. I don't do a ton of them, because I think most of the time we don't need to. But if you had a dog—so say you had a I know, like a hunting retriever type dog, some of those dogs, they might want to freeze 40 or 50 breedings and be in our office weekly for a months. Those dogs, I think freeze trials might be reasonable, because if he did freeze so much better in a different extender, maybe the freeze trial would have benefited you to do, but I think if they're freezing reasonably well, I probably wouldn't waste my time or money on a freeze trial. 

All right, so first step, after we've diluted them in, like our part one, we go into the fridge. So there are different types of refrigerators out there. Most of us either use an ice cream fridge, that's like a top loading fridge, or use a regular, standard, old school, you know, the white fridges that we used to have in our houses before the fridges all got really pretty and fancy. We'll wait for a period of time after they're loaded into the fridge. They really are all individuals. So some of these extender protocols might have a very specific window that you're supposed to hit. Two hours usually is minimum, because they need time to equilibrate to the temperature. That's the idea. Some extenders, they might do better the longer they're in it. So one of the notes that local therio person that I work a lot with, Dr. Colin, he and I both have appreciated that the Canopus tends they don't like to be in the extender for like, three hours. It doesn't say you can't on the instructions, but we noticed that a lot of these dogs really like being in an extender for six or eight hours sometimes. So just FYI. 

We're going to talk a little bit about pellets as well, but this will be a little bit focused on straws initially. Straws is how I package personally. They're both perfectly reasonable ways to go about things, depending on the equipment and your setup and who trains you to do freezing as a veterinarian. So semen and this scenario, it's drawn into straws. The straws are a half cc size usually is what we use as a canine industry. So we draw the semen, seal the end. All those straws are individually labeled with the dog's information, and we seal either with a powder—you pack some of the powder in the end of the straw, and then get the end of the straw wet so that it seals it. Or you can use these little beads. The beads is what I like. And then we'll arrange them on this little rack on the right. That rack, basically—I'll show you some little diagrams—but it's designed where the styrofoam part will float on liquid, and then that holds the rack higher than the liquid, so they're going into vapor before then ultimately going into liquid. So we want to make sure all the straw is super clean from surface liquid. Since it's going to freeze, it'll freeze water, right? Like the temperatures we're working at are going to freeze water, and we don't want condensation. Sperm doesn't like water, right? So good to try to keep everything as dry as possible. These aren't loaded with sperm, but this is just to kind of show you what this looks like. So each one of those little, like little grids on that rack, those could all hold straws. So you can put about 50 straws onto a float rack like this. So this is like a little diagram of how this works. So basically what we do initially, so we have a container, like a styrofoam container, or something designed specifically to hold nitrogen, kind of temperature. The styrofoam containers at Walmart work great. That's what I use. We put nitrogen in the bottom and let it equilibrate. So initially it'll be super hard to see in the gas that's being produced by that liquid nitrogen, and that vapor will cloud your ability to see everything down there. But once that's stabilized and we can kind of see the liquid at the bottom, we'll measure it, make sure it's three to five centimeters, a couple inches depth. That's going to be enough for what we're doing. And then once that vapor cloud is steady, we set that float rack in there and let it for, usually for 15 or 20 minutes. The second steps: we then take straws and we dump them in the nitrogen, so they have a period of time where they're above nitrogen in the vapor, and then a period of time where they're in the liquid.

44:05

But ultimately, the temperature that these are, so you know what we're working with, it's -320 degrees Fahrenheit, so it is extremely cold. If you touch this liquid, it will cause third degree burns. So we have to be very careful with it. You’re supposed to be handling it with gloves. I would say most of us, just our hands, are the casualty. Over time, if you're working in the vapor, it'll start to make the ends of your fingers a little bit numb. So alternatively, if you were doing pellets, it's a little different. Pellets use dry ice technology. Essentially, they're pipetting small amounts of the semen into what's called pellets in these little notches that are created in a dry ice structure. I'm not certain, like, the fine details of how the pellet kind of process works. It's not something that I'm super familiar with. I've seen it done, but I haven't done a whole lot of it. Companies, I would say, that tend to freeze in straws, tend to freeze in straws, and the companies that tend to freeze in pellets, tend to freeze in pellets. It's just like who taught you. But technically, you could take this extender and freeze it into straws, or you could take the other, the straw extender types the companies make, and freeze it in pellets. They just don't tend to sell their products out to just the general population to do so. But they do perform, I would say, the same overall, like we don't see an exceptionally good post-thaw with straws over pellets, or pellets over straws. Again, I think a lot of that is what is being told as a post-thaw is just miraculously high for a lot of the companies that do pellets. But it's not that we actually have high post-thaws specifically for pellets. I should say too, so then they would go from this state they would pack these into. I'm actually not 100% sure if they go from this to nitrogen before putting them in the vials, but they'll eventually put them in vials. But they have these little vials that would contain the pellet. And then those pellets would go similarly on canes, like we would have in our straw kind of setup. It's a different type of cane. This is the next step. It is going to the tank. So I was just going to show you. This is one of those kind of nitrogen holding tanks that we often will have in, like, a really busy or high volume type clinic that's doing a lot of freezing or maintaining a lot of tanks. We'll have these delivered on the right. The tank themselves, most of the time they're these kind of, I would say like three foot tall tanks that we have on wheels that have different canisters inside, so the canisters all hold canes, but like the picture on the left, you can see there's a vapor film. So there's liquid nitrogen inside that tank, and then there's a vapor film. It's like when we were working in the styrofoam container to keep this all at a constant cold temperature. On the left, you can see there's like six different hanging things that are little metal pieces. If you pick one of those up and you kind of move it into the center column, that is exactly the size of the column if you were to pull it completely out. We don't pull them completely out, obviously, because there’s semen in there, but we have all these little labels on the top, and this will be the same. This is the same way it would look if it was pellets or straws. It's just the way that we organize these tanks and know where we are. So if we say this is going to be tank four, canister one, and then we look for his name and date, and that's the easiest way for us to find it. So I know we already talked a little bit about this, but that is what the pellet assembly would look like. So they'll have these pellets in the container. We have to—with our bare hands—they will tell you to take these off with those cryopreserved, you know, Michelin man gloves, but these are tiny. These are not large vials, right? Those little pellets are teeny, teeny, tiny drops. And so you basically have to use your raw hand against something that's -300 degrees temperature-wise, that will burn you when you're handling it. It's not fun! But that's basically what we have to do in order to use these vials appropriately. Unless you're using tools to handle the ends, like, using the forceps, like what this is using and trying to unscrew, depending on how tightly they screwed on. If there's nitrogen loose in these containers, sometimes there is, it's just a little bit of a mess. Again, no study has shown one to be superior in breeding success in post-thaws and whatever your preference is. The only species that use pellets: goats, people and dogs. Everything else uses straws exclusively, pretty much. I like to use straws for a number of reasons. I was trained on straws. You know, like I said, one is not superior to the other from that perspective. But as a veterinarian, it's more consistent. I know what the volume is. We'll show you some examples of pellets that was not ideal. I know the semen is not going to get lost on the bag that we have to thaw in, because that bag, those bags are big that we thaw those pellets in, and everything kind of cooked the inside, which was not ideal. Safety during the thawing process, I'll show you some little diagrams on that. And like I said, it's more common industry wide. I would say more people are more comfortable with thawing straws than they are with thawing pellets, unless they are a pellet facility. I also feel like some companies that have pellets will not send them to us like this. They will send us a giant vial. If we ask for four breedings, because we want to use two breedings at a time, because that's the client's preference, and we breed her twice, they're going to send us one giant vial and just be like “All right, well, figure it out” and give us no equipment to try to separate these out or do anything. Meanwhile, the straws are easy to use, like you cannot screw up the straws, if that makes sense. It's like a monkey could do the straws. 

This is a real example. This is my picture. A client with a Tibetan Mastiff that sent me semen and said, “I have semen from the XYZ company, and I got 10 breedings on my Tibetan Mastiff before we neutered him.” And I said, “You could not get 10 breedings on your Tibetan Mastiff before you neutered him. As you probably know.” She was a scientist. She understood, you know, so she wanted to have it assessed. So we looked at them. We've had some nitrogen out. We put the semen in this little foil kind of apparatus that we made. I know you can't really appreciate the detail, but these are discs. They're not spheres, first off. The other thing is, like, look at the size difference. Like, look at the size of one versus another next to itself. And they say on the paperwork, there's no information for us all. All it says is: “One breeding is six pellets.” Which pellets? is my question. Also like, if you're a math person, you know that we're dealing with spheres. If it's double the radius of the other one, double the size in width, it is eight times the volume. So, big problem, right? Big problem, and estimate how many pellets to use. That is very important versus I think there are companies that do a much better job. Like, look at how different those are. Can you use a micro pipette to, like, pipette out exactly how much you need? Yes, you can. Some companies just don't do it. They either don't do it, they don't realize that this is their product and that this is what we're seeing on our end, but we don't like seeing this on the right, right? And we already talked a little bit about how it's tough to handle those vials. So the straws you can pretty well grab. I mean, I'll grab a straw with my bare hands, right? I will, but it's easier to hold them and not have to touch them all over and try to unscrew them. If we're using these big old gloves, it's really tough to be able to get the vials off of their canes. So the discussion is like, okay, we're thawing these pellets in a little bag. We're supposed to aim this vial over a little bag, but we're in a water bath, and the water bath will dissolve the sperm, and the sperm is gone if it falls outside of the bag, right? The straws, we're just throwing straws into the water bath. They're already in their own container, right? So if you drop the semen on the ground, if you drop the vial, because we're humans and like clumsiness happens, if you drop the vial on the ground, hopefully, you can collect whatever is left in the vial and throw it into the bag as quick as possible. If a straw falls on the ground, we pick it up and we put it back in the nitrogen. It's really, I mean, you got nine seconds, technically, in the air before anything bad happens. I don't think we could play pickup sticks with the pellets there. And then again, water is toxic to sperm. You missed the bag—that semen's gone. You drop the vial in the water—the semen's gone. You have a hole in the bag—the semen's gone. Like some of these things, we just don't know initially.

So this is the math that I'm doing that you guys don't have to worry about, but I just send you a nice report that says, like, “This is how many breedings you have,” but just so you kind of know where that comes from. So this is Zach. He's a young Dalmatian. He collected wonderfully, 800 million sperm, and it was 90% progressively modal—fabulous sperm as pre-evaluation. So what we're going to do is we're going to dilute his semen, so we might centrifuge it first, but we're going to dilute down. We put it at a concentration of 200 million per mil. So that's 200 million in a milliliter of fluid, which was just like how we began concentration terms. The total number of mils that he has is four. So if you can do the math, four times 200 is 800. That's how we get kind of back to 800 million total. All right, so we've got four mills. One straw holds half of a mil. So we have eight straws—is what we need to make up. And we're gonna make labels for eight straws. We're gonna get eight straws ready for this dog, and that's going to be 100 million in each straw. Remember, he had 800 million to start. Post, he thawed at 50%—fabulous. Next step, how many sperm? How many good sperm are in those straws? If he posts-thawed at 50% then if there's 100 million in a straw, half of them are good, then he's got 50 million in a straw that we can breed with. If we want to use 150 million modal for breeding, then we're going to use three. If we want to use 200 million, we'd use four, right. And then if we go back, so we had a straw that we use for a post thaw or partial straw, and then we end up with seven straws left, right, because we use one. Eight straws to start with, seven straws total in storage. Breeding three straws. So we divide seven by three, we have 2.3 breedings. That is a good collection from a Dalmatian, keep in mind. With a moderate post-thaw; that wasn't like the gangbusters post-thaw. But you can see how that math works. 

So red flags: let's start with this red flag. Your dog has 15 breedings from Dave's collection. They were amazing. Their post-thaw is 85%. That is super, super common, super common coming from a certain company that loves to come to dog shows in my area, and is quite literally, the reason why I freeze at dog shows, because when we receive that semen, it is super dilute. It is not 85% post-thaw, because fresh semen generally isn't 85% modal. I mean, half the time we don't even have 85% modal fresh like, how could we expect them to go through a freeze process, be frozen, brought back to life, and still be that high? It's just not a thing. So, like we said, exceptional, exceptional freezing dogs, we might see numbers like six, seven, eight on a special circumstance, but I'd say average, most dogs: two to four, maybe five, if they were really gangbusters collectors for their body size. That's really more common. Some dogs, we are going into it knowing we're probably going to have one, and that's very much worth it, depending on you know what your goals are for that dog. It's just realistic. We don't want to breed with crummy, dilute semen. We want to breed with something that we feel confident is going to get her pregnant. That is the goal. 

Other red flag for if you find a place that you're going to freeze at: if they say our semen doses are proprietary for any information, run. Run away. It's not helping you. It's not getting bitches pregnant. That is classic, I don't know. It just drives me crazy. It is science. We are trying to get them pregnant. This is science. We want your bitch to be pregnant. They should want your bitch to be pregnant, whether or not we know what their doses are that they have constructed out of 50 years of, you know, crap. It's just asinine. It makes me really upset whenever they send those to us, because they'll send us this. They will send us on our paperwork that we receive as the veterinarian with nothing but the post-thaw percentage, and this is one file as one dose, and that is all we get to have. It's just so frustrating. Whereas, you know, if I send semen to you, I want you to know everything that you have. I mean, if you're buying the semen from somebody that I froze semen from, I want you to know every morphologic detail. Like, I want you to know what you have. And if your vet wants more, then you guys can have that discussion. Like, instead of just having a scope in a dog and thawing the semen and going, “Well, this sucks, because this is we're here and there's no more semen that we can go grab. So, bummer.” Also be aware of what shipping costs. So again, a lot of these companies . . . I mean, I just had Cabana’s sister had a quote from a company on the East Coast for $1,200 for a non-time-sensitive domestic shipment of frozen semen for her bitch to be bred. And it's not that you can't charge $1,200 to do that. I'm not in charge of those companies, and like, you know their time is worth time, but it costs a lot less to ship from some places than other places. So like, you should be asking those questions, because if you don't live over there, it's going to cost a lot of money for you to get it back. But does it make sense for you to freeze with that location? Maybe not. Sometimes it's like hidden fees that they don't talk about. A lot of it's like they'll put a sign up and say, “Oh, it's only $300 and $200 when you come back.” But what they don't tell you is they're going to mark up after four breeding units—thats their breeding units, not your breeding units. If they say that your dog froze 18 breeding units, they're going to back charge your credit card for $1,000, but it feels good to have, like, this gargantuan number, and you never have to freeze them again. And so like, people are okay with it, kind of. Some people are not, but just watch for that kind of stuff. 

All right, so we're going to kind of skim past a lot of this storage stuff. So basically, the storage could be on site or off site. There’s plenty of places you can store. I mean, sometimes we'll collect semen and the client will pick it up and store it on their own properties. People that have cattle or, you know, if they have a tank that they already maintain. Usually easily facilitated within a couple of days. Most major companies will need a couple of days to get it out. Some of them can get something stat. But if you have kind of a mom and pop storage like mine, I mean, I can't get semen. I need a week’s notice to charge a tank to get everything ready. So just make sure you give them time. Most of us would much rather have semen way ahead of time as a backup plan, even if it's a backup semen plan. It depends on site. You happen to have a vet that is freezing and storing on site, well, that makes that super easy, right? Only becomes challenging if that clinic changes course. That has happened to some area clinics in my area, which is a little bit hard for some of those clients, if they're paying $1,000 a year to store semen over there, and they no longer have the vet that they use to freeze and do all of their breeding work with, little bit complicated. So this is just what the shipping tanks look like, just so you know. So they have this outer sleeve, they've got padding on the outside, and they got this little tiny tank that's like a little tiny version of what we have for storage. That's what will load things in. These are good for a couple of weeks, sometimes out to three. It kind of depends on the type of tank you have.

59:32

So if your dog has a fatal accident, gets in a dogfight and is dying at the ER, bloats. I mean, all sorts of things can happen, right? And I know it's not something fun to think about, but those things happen. I mean, I have clients all the time. I had a client last year that had a dog that died from a disease called pythium, and she was trying to find this procedure and it didn't work, you know, with the facility that she was in and how long the dog had been under treatment or under illness, but had a freak infectious disease, and he was a young, healthy dog, and she swore at that point, you know, “I'm going to make sure I freeze all my dogs when they're a year too old, because I would have used this dog in my program. He's the best dog I've ever produced and died from a, you know, freak incident.” Well, it's nice to be able to have something to where we can freeze some semen in some of those scenarios. 

So there's a procedure called gamete rescue or epididymal flushing in males that can be done. So this is an equine testicle, so it looks a little bit different than what we would look at—it's mostly like the size of the suture compared to the animal things. And you'll see my hands as an example of how the size is not right, right? But makes it a little bit easier, kind of being able to see all the details. You can see the testicle is kind of the big, round structure, what we think of. And if you think about when you feel your own dog's testicles, if you think about the knobby part that's on the bottom, usually, should be—that knobby part is the part that is the epididymis. That is where the ejaculate is before it comes out of the dog when we're doing a collection. They can store days’ worth of sperm in here, right? So if you collected him today and collected him tomorrow, the reason that he still has plenty of sperm left is there's actually quite a lot of storage in there. So that is where we harvest; that epididymis is where we get this sperm from. So what we have to do when we're doing this process is we have to neuter the dog essentially, or, you know, rough castrate the dog if he’s deceased. We will take all the accessory tissue, so we strip all of the vessels, all of the kind of extra stuff that's associated with that epididymis, detach it from the testicle. And you can see on the first picture, we're just kind of starting on things, and then you can see that, like the little guy that looks like a brain, you can see he's no longer attached, right? So I've removed him from the testis itself. Again. This is a stallion. This is not a dog's testicle. This would be a giant dog testicle. We removed the testicle. We continued to dissect. We rinse with saline and try to clean everything off. And then eventually we have this super clean version of what we're trying to work with. We'll put a catheter. So in this scenario, this is with an equine sample, we are flushing using a Tomcat catheter, which is a much larger gauge. We'll use, actually, IV catheters—is what I'll use to feed into the tubular part of that structure. And then I'll usually make some little cuts with a pair of scissors, like some sterile scissors in the epididymis itself, so the part that looks like a brain. And then we will use extender or saline to flush backwards through that tract. So basically, we're putting it through a reverse of where it should go in normal anatomy. But it's a little bit easier for us to get a catheter in the big side, if that makes sense. So we're able to flush semen out through and into a dish. I'll sometimes just flush it into a conical tube, just kind of like what we use in the clinic anyway. For this procedure, we have to get testicles essentially within 12 hours of death. So if you have a dog that you have at 10pm at night that you back over in your driveway—I know it's not fun to think about. I've had these clients call me. What you should do, you don't have to do anything emergently. The ER isn't gonna be able to do anything if the dog is deceased. The dog is deceased, and you could go and confirm, certainly, if you're not confident and assessing that he has died from whatever happened to him. If you think his testicles, based on his health history, his age, etc., are reasonable to pursue this procedure, I would put that dog in a refrigerated setting if you have the option to, even if it means an ice chest and laying him on some ice and an ice chest, that is a reasonable choice. It's better than if he was room temperature. And then pursuing some of these things in the morning as soon as they can be done. So you can do these castrations a long way out. I think 36 hours is the latest I have done this, based on a shipping timeline. So, like, the dog died on a Sunday, couldn't ship till Monday. The vet that had the dog didn't have the ability to be able to do this himself, so he shipped it to me. And in that scenario, fortunately, basically, we were able to retrieve semen on that dog, even though it had been almost two days by the time we actually did the freeze process, which is amazing, and it's so exciting to be able to do these procedures. 

So basically, the other way we might use this . . . so there are some dogs, they might be alive at the clinic. I had one that was a third time offender, GI foreign body, and I could not fix his GI foreign body the third time. His intestines were too far gone. Well, that dog I was able to neuter as part of his surgery before we euthanized him on the table. So, like, that kind of approach is possible. And then every once in a while, I’ll have a dog that's there for a routine neuter, but say the owner had a fleeting thought, and the dog is terrified of being at the clinic, and they don't think we're ever going to be able to figure out how to get semen collected on him because of his behavior—sometimes it's like livestock guardian dog, things like that, where there's just no way that I'm going to be able to condition that dog to collect semen. I think it's a perfectly reasonable choice to do this as part of a kind of a post-neuter plan. I've also had, like, I had a Belgian Sheepdog, one that was really difficult to collect, and basically, I could collect, like, 200 million sperm—that was about all I could get from him. And we did that at the end of his . . . he was going to a pet home. They were trying to collect and bank semen on him, and they just were thinking, “We'll have as much as we can. So let's do an epididymal flush when we finish with his attempts at freezing. The last time we attempt it, we'll just plan for his neuter and do a flush, get as much as we can.” It's an expensive approach. I mean, this is not cheap to do, right? It's a very specialized procedure. So I think for most people, that's not going to be a feasible option. But you know, if money is not a concern, it's certainly a possibility. 

So this is just my PSA to say: this can be done. Have a contingency plan. Talk to your regular practice. Talk to your regular ER, if you have a relationship with an ER. Call local universities or local therio people. Sometimes, I think universities, it tends to be the best option, because they usually have 24-hour call for emergencies. They usually will wake up their residents with questions. I know because I've been there. And they tend to be really excited to do this type of casework. This is a really fun—I mean, I know it's not fun for the owner, but it's a fun task for a resident that is learning how to do this kind of thing, to be able to get hands on experience doing those, not at cost of quality care, but it's nice to be able to have somebody available all the time. And we do learn how to learn how to do these when we're in our residencies. So any vet can remove these. You can send them overnight. Somebody who can do it. Just have a plan of how that might work, or what university you'd work with, or what practice you'd work with. So I do this every once in a while. I think it just depends, like, do I have enough nitrogen on hand? Am I about to do a freeze for another freeze day? And I know that I'm not going to have enough nitrogen to do what I'm about to be scheduled to do for freezes that day. Well, I might have to turn you down if I don't have enough nitrogen in my container. Sometimes I think people grasp at whatever they have available. There are companies, again, some of the technician based companies that are out there that are encouraging people to ship semen to do this. I don't think shipping semen for you doing this is best if you have a local option. And I would be very scrutinous of the companies that are offering this are not usually the companies that I would recommend storage, freezing, etc., from the get go. So just be judgy and keep open minded about the fact that maybe the people who are going to be available might not be the right ones to be able to do this for you effectively, even if they claim to. 

So I know we ran over. I was just going to let you guys know if you are in my area, Genesis is my personal freeze business that I have. And I just was going to say if anybody has questions about freezing or, you know, dogs that you're looking to freeze, or things that you're needing to do, I do have a freeze show scheduled in a couple weeks, so I'm going to be really absorbed here soon. But I just want to mention, if you're trying to find me in Dallas, I am at either Animal Medical Center Richardson—or at Cohen Veterinary Reproduction is actually opening a clinic in December that I'll be spending a lot of time at as well. So if you're trying to find me, that's where I am. If it's for other things. If it's freeze related, I'd say the Genesis contact is great. So I will take any questions, and I'm happy to stick around for the length of those. I'm sorry I'm terrible at shortening these to an hour. It's just one of those things, right? I get really excited.

Nicole Engelman  1:08:59

We love excitement about a topic. Thank you so much for such an in depth presentation. I can tell everyone has absolutely loved it. I want to be conscious of everyone's time, so I'm just gonna ask a few questions that we saw come in. We had some people wondering about motility percentages, and is it typical to see motility percentages drop so drastically upon thawing?

Dr. Andrea Hesser, DVM, DACT  1:09:22

Yes, so I would say it is more common. What has been trained into belief is that dogs routinely post-thaw at 80 or 85% and they don't. They don't. When we receive samples, I would say generally, it's a really good freeze if they freeze at 70 or 75%, and it happens. I have it happen all the time, but I would say it's probably 20% of the time. I mean, it's not a lot of the time. I'd say more commonly, we're in the 60s or the 50s, but I would say minimum, most dogs will drop in post-thaw at least 10% from where they started. And where most of these guys start, I will rarely call a dog 90%. I know that was my example, but I'll rarely call a dog 90% progressively modal. I'll usually be in the 80s to start with. So if you think about it from that perspective, that is the best case scenario. Outcome is the 70-75%. There are plenty of dogs we post that are 30% modal, but if you do the math, it still ends up with a certain number of breeding, you know, depending on how much semen they collected. We just don't know how they're going to freeze until we freeze them.

Nicole Engelman  1:10:28

Great. Thank you for answering that one. I know we talked a lot about extenders throughout this presentation. One of our breeders was curious about your thoughts on females failing to implant or carry a pregnancy as a result of extenders.

Dr. Andrea Hesser, DVM, DACT  1:10:44

I think this is one of those, “Could it be a thing?” I think it's not impossible that a dog has . . . so even things like, think about anaphylactic reaction. Like if you had an allergic reaction to eggs, if you're allergic to eggs, well, frozen semen is going to be a real challenge. It’s going to be a challenge. You know, most of these extenders are egg-based, so it is possible. There is no reason to be outwardly fearful. I've never had one that I felt like that was the thing. I just would usually say, we don't have enough tries in this species. We don't have enough cycles to work with that we really can predict a pattern. On like a horse, for example, we might breed her with frozen semen today, and if they don't get pregnant, she's cycling. We're going to breed her again in three weeks with frozen from the same sire until she's pregnant with that frozen. We're going to keep doing that. We don't get to do that with dogs. Usually, once we have a failed frozen semen breeding, we go, “All right, well!” That’s just like with my dog. I'm going to breed her with fresh next time, because I want to make sure that I have something to go forward with. I might breed her with frozen again, but I'm going to switch it up now. I think that's the downside. So maybe that's why some dogs don't get pregnant with frozen. Maybe that is the reason. But I think the reason people are afraid to invest in frozen is because what we've been doing for years, unfortunately, is expecting good outcomes with really crappy semen and really poorly packaged semen. And sometimes it's not in your control as the owner of the female dog. You know, we're talking about this because we own both ends of things, but usually you don't have control if you can get free vials from that facility, because the stud dog owner might not have any idea that he can't have 15 breedings from his labrador from one collection, and he will not give you more than one vial. It's tough. It's a tough position to be in sometimes.

Nicole Engelman  1:12:32

And just kind of in that same vein, with extenders, what is a point that you would say it's worth breeders trying to use a different extender, if they're just not seeing the results that they would want to? Would switching the type of extender they use make the difference or not really?

Dr. Andrea Hesser, DVM, DACT  1:12:49

If I have a dog that has an exceptional appearing collection, and we go to freeze it, and it freezes at less than 30, I will bring it up as an idea, because I usually use the same extender. If they don't like my extender, well, maybe you go to another practice that's in my area that freezes and stores. So they can use a different extender, because they use a different extender than I use, and just see if it goes differently. If it goes differently, great. I have had weird stuff that I couldn't explain, that I think there's more to it than just the extender. I think sometimes it's situational. Some dogs are easily cold shocked. I think sometimes it's like the equipment that one person uses over another, the time that they're in extender, one versus another. A lot of times, if I'm collecting a dog frequently, I'll look at all those details. If I have something weird that happens, I'll assume that it was me. It might have been the dog, but often I'll tell that client, like, “I'll just comp the next. Like, bring him back to me, and I'll freeze him again, on the house,” just to make sure that it's not something that happened. Because you saw all the details on this. There's so many things that you can do that will kill it, that if you didn't do it right, or the timer didn't go off, or you got distracted, or, you know, things might not thaw well. And I think that's what I would say over extender individuality. But I do have some dogs that—we will freeze them, and they'll freeze differently with us than they do at another practice with a different extender. And that's perfectly fine if they do better in one. I'd stick with the one that you're getting good results on, obviously. 

Nicole Engelman  1:14:19

Great. Thank you for answering that one. And then we did have one quick clarifying question, if you could spell and repeat the name of that extender that you had recommended?

Dr. Andrea Hesser, DVM, DACT  1:14:31

The one that you have to make from scratch is called Uppsala. It is actually written, if you're looking at the spelling, and you look at the rewind, or look back at the presentation, that little graphic in the box, I think it's like the center of that slide has it written as well, just in case. So it's actually a university—I think it's in Sweden, I believe. That’s just what we call it. It doesn't have a name, so we just call it the university's name.

Nicole Engelman  1:15:00

Oh, okay, amazing. Thank you for clarifying that one. And then I think we have one last question here from Christine, who is curious if there's a list of storage facilities in the US that breeders can access. In other words, they use a vet at the facility near them or near that facility, basically just looking for storage facilities near their vet. So perhaps it wouldn't even need to be shipped.

Dr. Andrea Hesser, DVM, DACT  1:15:23

Yeah, and that's a good question. No, there's not a database. There are companies that have centralized storage options. So there's pros and cons to having semen at a clinic. I know we kind of touched on this, but the pro is it's there for you. The con is they’re a private practice. I think the con lies a little bit in safety, truly. So some of it is the person that was in charge of that tank, taking care of it, or did they get pulled to surgery when they were supposed to check the nitrogen levels because somebody had a C-section, and then somebody gave them a phone call, and that happened a few weeks in a row, and they had a tank that went dry and they didn't notice it, and then all that semen gets lost in doing that. That is the nature of veterinary practice. We're busy people, so it is tough to devote time. I know some clinics will have safeguards around that, or have a separate person that answers the phone for that kind of stuff. But that can be the challenge with that. Meanwhile, the perk is, you know, you can use the semen on site. In contrast, a lot of people that are like–Zoetis is the example that I use because I'm familiar with it. Zoetis has 50 veterinarians around the country that are freezing, generally with their extender or with their protocol, but they have been trained to, like, know how to use all that kind of stuff, streamlined and do a good job with it, and had veterinary oversight on top of themselves, being veterinarians. Those facilities will ship back themselves, like, if I wanted to use semen on the dog that I have stored in Zoetis, they will ship it to me for $100 including the tank rental, instead of $1,000 from another company. They will ship it for almost nothing. Like, I don't even know how they do this, because, like, if you went as a person, went with your 30 pound, three foot tall tank, to ship it, it's $375 or something like that to ship overnight from one place to one place in the domestic United States during the weekdays. So I have no idea how they managed to do this, but they basically, for all of their clinics, they do all of the storage. So local clinic that does all the Zoetis training in the country is right around me. They don't store any semen on site. They have all of their semen at Zoetis and have been freezing semen for Zoetis for 20 years. All of those clients, they want to use their semen it’s $100 to move it back to themselves. There's a lot of clinics that will charge $100 even though you don't think you're going to pay anything that you are paying something for them to go and figure out the paperwork and pull the semen out of storage, move the semen into a tank, move it into surgery. They might charge you 100 bucks to do that. It's not ideal for like—we don't want to ship semen around if we don't have to, but I think it's a very good alternative. It just has a different list of pros and cons. So sorry, that was kind of a wordy answer to that, but hopefully that was great.

Nicole Engelman  1:18:15

Thank you for answering that one. I think that was our last question that we have time for. But Dr. Hesser, we cannot thank you enough for doing this presentation for us. I think everyone just learned so much. We love having you here, so we'll have to have you back again sometime very soon. So thank you and thank you to our audience for joining us today. If you're not yet a member of our community, you can apply to join at GoodDog.com/join to stay up to date on all of our future webinars and events. We still have some more coming up to end the year, so we're really excited for that. And until then, I hope everyone has a wonderful day and we'll see you back at our next webinar. Thanks everyone. 

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