Afghan Hound Times -DENMARK 1997 3rd World Afghan Congress Report #7 Wendye Slatyer (AUSTRALIA) "FROZEN SEMEN - advantages, disadvantages, methods, ethics and technique"..

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DENMARK 1997
3rd World Afghan Congress Report #7
Wendye Slatyer (AUSTRALIA)
"FROZEN SEMEN - advantages, disadvantages, methods, ethics and technique".

Acknowledgement: We wish to thank the author, Wendye Slatyer of Calahorra Sighthounds, for permission to re-print this article which was prepared for the Third Afghan Hound World Congress, Denmark, 1997.

Introduction and background.

Firstly, I would like to make it very clear that I am not a veterinarian and I realise there may well be people here who have a much greater depth of knowledge as to the technicalities of this subject than I do. I am speaking today because our kennels pio neered this field in 1976, so we have been involved with canine frozen semen literally since its beginning, with our latest success only last month. We were fortunate to live in Australia, the country to first recognise this invaluable breeder's tool and to encourage its extensive utilisation.

If there are some specific technical aspects which I do not cover to your satisfaction, then hopefully some veterinary experts who are present here today can answer them for you from the floor.

I also apologise for using some of our own dogs' names. There is no intention to promote our own kennels through this Congress, but some details are essential if parts of this discussion are to make sense.

So to some BACKGROUND

The first time in the world puppies obtained from AI by frozen semen were registered with a Canine Controlling Body was in 1976 when a litter of four was born at our Calahorra Kennels, from semen we imported from England to Australia.

We did this because at that time Australia was closed to live dog importation due to a rabies scare in England, and dogs from any other part of the world had to go there first for a period of twelve months before continuing their quarantine in Australia. Should the ban not be lifted, it was vitally important to find a method of ensuring that bloodlines could continue to come in to Australia. Hearing that some puppies had been produced under clinical conditions by frozen semen at Cambridge University, we d ecided, with the help of two famous veterinarians, Dr Harry Spira of Australia, and Dr David Morton, then at Cambridge University, now in Leicester, to ensure this became a reality. We selected Ann and Jenny Dove's English and Irish Champion Saringa's Abra Cadabra, as his bloodlines tied in very well with those already long established in our breeding programme, we had seen him on a recent trip, and considered him an excellent repres entative of our interpretation of the Afghan Hound.

Of course we were not the only people concerned by the ban, so semen for some other breeds came in the same canister and our litter was closely followed by Golden Retriever puppies and then a long time later a single Old English was born, with subsequently a few other successes in various breeds.

That was how it all began and now the whole world is involved.

Until recently in Australia, semen could only be imported from dogs resident in the UK, now it can come from almost everywhere. Considering our quarantine expenses, this has turned the Australian dog scene upside down, especially in rare breeds, and overa ll the successful development of artificial insemination from frozen semen has been a wonderful tool for breeders.

HOWEVER, MANY PEOPLE STILL ASK - "WHY USE FROZEN SEMEN?" What are the advantages, what are the disadvantages? How is it done? What ethics are involved?

So let us first look at the ADVANTAGES, which in our opinion far outweigh any disadvantages.There are many major reasons for utilising this technology and more are emerging as the success rate inreases and more breeders decide to incorporate it as a regul ar tool in their breeding programmes.

The primary most obvious one is being able to source what you consider the most suitable material for your breeding programme from almost anywhere in the world. You may choose him because he is very famous, or you may come across him sitting in someone's backyard, virtually unknown but ideally suiting your specific purpose. The greatest advantage lies in being able to directly access that SIRE himself, rather than having to buy a son, hoping that he comes up to expectations. Even if he does, he is STILL o ne generation further removed from the actual dog that interested you in the first place. Only very seldom is the selected dog himself for sale or, if so, affordable.

At the same time, using frozen semen makes it possible to overcome geographic isolation and avoid the many disadvantages associated with quarantine which of course are all too well known.

Matings that could otherwise never occur can now take place. Properly used, therefore, its value is inestimable.

For example in the past twelve months I have had the pleasure of judging what I consider to be two outstanding Afghan Hound males, and immediately decided that I wished to incorporate both of these top winning, stud dogs into our lines. Both are bred-in- the-purple and pedigree-compatible with what we have been breeding for years.Without frozen semen, there is no way it would be possible.

One was in America, the other an American import in Italy. One carried the same Old American and Old Scandinavian lines with which we have worked for years, inclduing many of the famous individuals, the other is less directly related but rather through f amily lines with the same phenotype and genotype we work with consistently. Having seen many of the direct forebears of both these dogs in the flesh, we feel they will tie in perfectly with our own blends of some of the world's greatest bloodlines. We imm ediately arranged to import semen from the dog in America and one black masked silver bitch puppy was born a week after I left Australia, and as you can imagine, I can't wait to get home to see her.

To my delight the owners of the dog I loved this trip are also happy to provide semen, thus enabling the perpetuation of his gene pool in far off Australia, via bitches who could otherwise never be mated to him. We have already collected from him and hope to obtain puppies before the end of the year.

I have also been very impressed during my recent judging assignments in Europe and Scandinavia with the fact that so many of my winners over a variety of breeds have been produced from frozen semen, including Best Opposite Sex at the Italian Afghan Hound Specialty. It is not always possible to recognise this because there is no indication in the catalogue, and it is recognised by knowing that a certain prefix belongs to a particluar breed in a different country, whereas in Australia, the letters A.I. MU ST appear after a dog's name. They are part of his registration and will always be seen whenever his name appears, in catalogues, advertisements, service certificates etc. Personally we feel this is most important and it is regrettable that other countries do not follow this procedure, but of course in contrast to our regulations in Australia, many do not include the country of birth in the registration details of imported dogs either and again we feel that this is not in the best interests of accurate record keeping.

Another reason for using frozen semen, and in our opinion the most important of all, is the long term storage of precious genetic material for future usage, thus greatly expanding the potential for serious and ongoing line breeding to famous stud dogs.Thi s also allows for practical evaluation of contemporary stock by comparing the results of these litters with those produced only from current parents. Unfortunately we all know that show criteria are subject to fads and fashions, despite the careful wordin g of the breed standard. A breed can change greatly over a few short years, and not necesarily for the better. Long term stored semen therefore presents an invaluable opportunity should you wish to go back and re-incorporate the valued - sometimes even es sential - traits you may otherwise find hard to source.

As some people have a problem with the concept of puppies being produced long after their sire is deceased, their concern will be discussed a little further on.

Carefully pre-planned line-breeding from long-stored semen is an area with which we personally are currently deeply and very successfully involved. Once again we were the first in the world to achieve success.

You can see here a painting which includes three of our most famous stud dogs, all long-deceased. Since 1992 we have had an intensively line-bred litter from each of them.

In 1992 we produced a single pup from semen that had been stored for sixteen years, and he quickly became the 40th champion for his sire, our long deceased Flying Dutchman of Isfahan (imp USA), who was born in 1968. (Second from left in the painting.) A Multi RUBIS winner, this dog born in 1992 has already sired three champions including Championship Best in Show and Runner Up Best in Show winners. His dam was Ch Calahorra Fledgling, who was herself line bred twice to Dutchy, via her fourth and fifth gen erations and who, as a bonus via her dam Ch Calahorra Rowena, was also a grand-daughter of our historic 1976 litter.

In 1993, came the litter using 17 year old frozen semen from Ch Calahorra Quetzlcoatl, who was born in 1970 and sired 17 champions during his lifetime (first on the left in the painting). The current progeny are line bred five times to Quetzl, once to his champion sister Quo, and ten times to their sire Chandhara's Emir of Gray Dawn, with at least 30 crosses to American, Canadian, Venezuelan, Ch Shirkhan of Grandeur. Their dam Calahorra Born Toperform is a Rowena grandaughter, being out of Fledgling. In a normal breeding frame it is virtually impossible to produce so many generations of sufficiently mature age to establish such concentrated direct line breeding, other than by resorting to inbreeding with father/daughter or brother/sister matings. In our opinion, these are seldom justified and certainly not recommended for any but the most experienced breeders with great depth of knowledge of their own bloodlines.

This litter has created enormous interest world wide, two are already champions, and four more are well on the way with Best in Groups and RUBIGs. Grandchildren are also winning at the highest level with several having multiple challenge certificates.

On the right of the painting is Chandhara's Emir of Gray Dawn (imp UK), born in 1967 and sire of at least 21 champions during his lifetime. His newest daughter was born in 1995 from frozen semen stored for19 years at the time of the insemination. Her dam , who is also Calahorra Born Toperform, traces nine times to Emir of Gray Dawn, so this puppy's "type" was almost certainly assured.

By collecting semen for freezing, you can not only store your own famous stud dogs for re-incorporation into your lines at the right time, but also by pre-planning with frozen semen of course you have the equivalent of an insurance policy. For example, fr eeze any young up-and-coming star you may have, but which at your particular stage of development you cannot use in your own breeding programme simply because he is too closely related to your females at this time. He may well turn out to be an invaluable link for you in twenty years' time. The same applies to an outstanding dog you may sell, especially if he is to go out of your country. Remember also that dogs can die prematurely from many unexpected tragedies or causes, not just old age, or an accident al injury can prevent him from mating naturally, so if you value his genetic potential, freeze him. Of course ethics come into the equation at this stage, so more about this later.

For these reasons also, preservation of semen may become a crucial factor in the development of rare breeds.

A further advantage - and in fact an inestimably important one - has recently been brought to my attention, and that is the situation which occurs in breeds with genetically inheritable problems, such as PRA, Cataracts, Hip and Elbow Dysplasia etc. Pre- p lanning and responsible use of frozen semen could assist significantly in the control - possibly even the eradication - of these problems. To take it one highly probable step further, it is feasible that - in extreme cases should the gene pool of non affl icted animals become alarmingly small - frozen semen may become relevant to the very survival of certain breeds, by ensuring future access to these clean sires. Conversely, programmes designed to detect inheritable diseases in bitches could be implemented by matings with frozen semen specifically collected from affected or carrier dogs.

In American Cockers for example, a certificate for eye clearance is not obtainable until eight years of age. At that stage, many males will be decreasing in fertility, or if they have not previously been used at all, will be reluctant to mate. By storing semen from a significant dog when he is young, say at around 3 - 4 years, then it would be possible to use such a sire after he has been cleared, the overall result being that semen can be made available to the entire world, which could well assist breede rs in their endeavours to eliminate such horrific problems.

So you can see that if you have a stud dog you value and you plan ahead, by using frozen semen you will again be able to re-produce his qualities many years into the future.

A recent development is one well worth noting. Our vet Robert Zammit now suggests that as a matter of routine clients have a Government Pathology Laboratory do a Leptospirosis and a Rabies test prior to collection. Then should the dog die, semen can still be exported to those countries which specify these tests as requisite for the importation of frozen semen. In England, for example, once a dog has died, his semen can NOT be used, a classic example being the almost criminal waste of the semen stored from Ch Khanabad White Warrior, due to Margaret Niblock's great forethought at the time. However, if there was certification that he was free of lepto and rabies when the collection was done, it could be exported to a progressive country like Australia that p ermits the use of semen after death of the sire, a litter could be bred, and progeny returned to England to continue the line. Makes you think, doesn't it?

THESE ARE THE ADVANTAGES, SO TO A DISCUSSION OF THE DISADVANTAGES

Without doubt - and believe it or not in direct opposition to the outstanding advantages we have just discussed - the major potential disadvantages are the threat to correct breed type, and/or to the genetic health of a breed.

When we first heard that America was establishing semen banks, we thought "What a great idea!" Now we are not so sure. How easy it would have been in 1976 to contact a single organisation instead of going through the expense and the months of hard work we had to do then in order to make puppies born from frozen semen an actual reality. I am told it is now possible to simply order from a catalogue of available sires semen that has already been frozen and is simply waiting at the storage facility to be ship ped to buyers, and that this can therefore be done without any consultation with the owner of the sire, or exchange of the so essential background information concerning type, pedigree, health etc of both potential parents. Certainly I was approached by an overseas facility not many years after our 1976 success to make my stud dogs available for this purpose, but of course declined, as providing semen which could be purchased in this fashion is beyond my comprehension. I have not bothered to confirm whether or not it ever became reality, but if it IS true, then the long-term prospects for a breeding programme based on such unresearched genetics would seem to me to not be very bright.

Frozen semen can be potentially misused if a new "look" suddenly attracts attention, especially if it is in fact untypical as is often the case, or it disregards even just a single one of the all important breed hallmarks specified in the Standard. An ex ample is the lengthening of back in order to increase the potential for open side gait, now a buzz word in our breed, but one not mentioned ANYWHERE in ANY standard for the Afghan Hound. Nature says you may "fluke" one or two eye-catching specimens that a re outside of the original structure for a particular breed, but if you want to perpetuate this so that it will breed on, you can't do so without altering many other parts of the skeleton, as Nature is determined to maintain certain proportions.

So the whole shape of a breed changes.This variation on a theme can then far too easily travel around the world and be imitated and rapidly re-produced in considerable numbers of puppies by the use of frozen semen in a fraction of the time it used to take for such a damaging factor to become firmly entrenched in a breed. The breaking down of geographical boundaries and the decreased costs of semen versus those of quarantine can therefore be as much a disadvantage as an advantage.

The same applies in those breeds afflicted with heritable genetic defects, for exactly the same reason. We have already discussed the enormously important advantage of being able to get back to clear sires by the planned use of frozen semen. But of course again it works both ways, and if a significant sire is widely used by frozen semen and at a later stage is - tragically - found to be afflicted, then the problem has once again been disseminated worldwide and in far greater volume than would otherwise ha ve occurred if he were only used by natural methods in his country of residence or nearby places from which bitches could be mated to him at no great expense.

Fortunately this is not a factor which poses a major problem in our own breed, as it is very healthy genetically compared to many others, but it is certainly something that must be born in mind.

Equally disadvantageous, as the sourcing of frozen semen becomes increasingly easier and the success rate more rewarding, is always the risk in all breeds of a "craze" to simply import semen from current top winners, irregardless of the compatibility of t heir pedigrees with the bitches to whom they will be mated. This is an extravagant exercise in every respect and from the breed's point of view, another area which is ultimately fraught with danger, especially when semen from several top winners is import ed and admixed within only a few generations, outcrosses on outcrosses. The most obvious cause for concern is that very little line breeding appears to be occurring, with not enough - or in fact none at all - careful research into the pedigree.

Every pedigree carries some true pluses and some hidden minuses. In the case of imported frozen semen, in most cases very few of the close relatives have even been seen by the person importing the semen, much less their having any personal involvement with or understanding of three or four generations behind that sire.

Certainly in the old days this was exactly the same when dogs were imported from one distant country to another, for example from England to Australia. Very few of us were in the position of being able to personally select the bloodstock which ultimately arrived after seven weeks on a cargo boat.

And we did not have the wonderful advantages of magazines, videos, faxes and emails, which we all just accept today as part of our lives. But probably for that very reason, all the difficulties of communication, and all the homework that HAD to be done, i t was essential to research the new addition in great depth. There was - and financially still is - no going back if we did not like what came out of the shipping crate. Personally I believe a great deal of the world wide acknowledgement for the quality o f the Australian Afghan Hound is due to the depth of commitment that went into the selection of these early imports and has been carried on to the present day.

Likewise, where the homework incorporating frozen semen IS being done, some magnificent specimens are the result. Only recently an Australian born Weimaraner produced from American semen has gone to the USA and taken his breed by storm, winning the Nation al Specialty and gaining his American and Canadian titles unbeaten since his arrival.

While it is not technically a disadvantage as such, the disappointment when a bitch fails to conceive after all the time, money and effort that are expended seems more devastating than when this happens with a natural mating, probably because frozen semen inseminations basically require more forward planning and more co-operation from a large number of people than does the average natural mating
.
Anyway, back to our old semen and how we came to have access to it.

After our great success in 1976, we received a request from Sydney University to collect and freeze semen for future research into long term freezing of semen. We chose five of our most influential stud dogs, although at that stage there were no real plan s for when it would be used.

In fact for many years we had trouble finding out if it was even still in existence, as by then the rabies ban had been lifted, the University had moved on to other challenges and everyone forgot about the procedure for a number of years. From our point o f view also, the devastation created by parvovirus meant that we would not risk our precious small store until the threat of this hideous disease appeared to be well and truly under control, so the whole project went "on hold". We were very lucky, because several major stud dogs in other breeds were also involved in this programme and the semen of only a very few can now be located.

Towards the end of the eighties various Australian breeders began to have success with recently imported semen in many breeds, and our veterinarian, Dr Robert Zammit, contacted us and said that our old semen had been handed on to him by Sydney University as he had been working on an advanced degree in Veterinary Science with Dr Ian Martin when our original insemination took place. We carefully assessed our bitches and decided that the time was right, as we currently have the best possible genetic pool for re-incorporating these famous males.

As a result, another step forward was made in the world's knowledge and utilisation of frozen semen. When it was collected in 1976, the "shelf life" of the semen was estimated to be around 25 years. We have now proven that AT LEAST 19 years is feasible an d we will continue to use our long-stored semen as the "right" bitches present themselves for these matings.

When the 1976 insemination took place, we opted for an inter-uterine surgical implant rather than risk reducing effectiveness by the alternate method which uses a glass pipette. In fact at this early stage there wasn't any real alternative as the advanced type of pipette being successfully used in Norway where the majority of breeding on the fur farms was by A.I. was not then available in Australia, and we subsequently brought one home for Dr Spira. He later used this on the remainder of the Abra Cadabra semen for another of our bitches, but - probably purely co-incidentally - that time we were not successful in obtaining a pregnancy.

The fantastic result, as we have said, was four puppies. The dam had previously produced a litter of nine, so this was in keeping with the advice at the time to expect half an average litter size

The pups were larger birth weight than we had been used to and grew more rapidly, thanks to their hybrid vigour, but their size at adulthood was commensurate with the bloodlines involved and so was their phenotype.

Interestingly enough, they also had only one head and four legs each, despite the dire suggestions from some sources that performing an artificial insemination was bad enough, but to do this using semen that had been placed in a glass straw, then frozen, then packaged into a canister with dry ice, then travelled on an aircraft for more than 24 hours, then been thawed and finally placed directly into the uterus of the bitch by a surgical procedure, was simply not "normal" and therefore fraught wih all the dangers of the unknown. Impossible as it is in the light of today's innumerable successes with frozen semen, several otherwise intelligent people genuinely expected the puppies to be born with abnormalities or deformities.

Now that success has been achieved with semen that has been stored for so long, many people are again having trouble coping with this amazing technology. They feel sure that deterioration MUST take place, or that some previously unknown disease will emerg e, a bit like what happens in episodes on TV about the Starship Enterprise.The matings we are doing with our pedigrees of frozen semen A.I. on line-bred frozen semen A.I. are simply beyond the comprehension of many people.

Anyway, back again to the 1976 litter. The puppies themselves were of good type and quality but not sensational show prospects, but then first generation from live imports is also often not quite what is expected, and the breeder in both cases must be pat ient and allow the time necessary to get to that all-important second generation.

This was definitely the case with us, and when in 1979 the pick male, Calahorra Anglo Saxon (AI) was mated to our famous Royal BIS winner and top producer, Ch Calahorra Requiem, the result was, genetically, one of the most important Afghans we have been p rivileged to own, Ch Calahorra Rowena.Her pedigree carried two lines to Dutchy and one to Emir of Gray Dawn.

Rowena not only produced seven significant champions, but eleven years later, in 1990, our top three winners were her children, and today she is still behind all of our currently dominant show and producing stock, including the three "miracle" litters we have already referred to, each of these incidentally produced by surgical implant. Therefore, if we had not been part of the pioneering work done with frozen semen in 1976, our gene pool today would be a very different one, as you can clearly see.

At the time, however, despite world wide interest in the technicalities of the achievement, the Australian breeders basically said "That's nice, but ho hum, we've gone beyond the English Afghans anyway, we're all off after the modern American type now,"an d as I have said, at that stage we could not import semen from anywhere but England.

METHODS AND TECHNIQUE

Of foremost importance is the selection of the recipient bitch - not just for her quality but also, which may surprise you, for her potential to conceive to this technique, and many times this comes down to the breeder's instinct that this bitch is "righ t". For no explainable reason, some simply do not conceive to frozen semen although all the tests indicate they will do so, but these same bitches regularly whelp to natural matings. When this happens, especially with proven bitches, it is all the more di sappointing.

Also, despite the greatest amount of knowledge and talent, it is still possible that the bitch you choose for the insemination may not be the right one for that particular dog after all. Most people know how often a less famous bitch outproduces "the show star" of the kennels, despite that star being given every opportunity to also be a great producer. There is always an element of luck involved in the choice of two breeding animals, no matter how smart we may think we are when we make that decision - with the costs and hassles of frozen semen, the element of luck seems to be even greater and of course this is increased if the sire is a total outcross.

The initial contact when planning a frozen semen litter has to be with a very co-operative stud dog owner. Then comes the collecting veterinarian, all the requisite paperwork for Government Permits, Canine Controlling Body, and veterinary certification in both countries, co-ordinating all the blood tests (some are before collection on the stud dog and others are before insemination on the bitch), organising the shipping in one country and the collection in the other, and finally total confidence in the ve terinarian doing the insemination. When ours says "ready" we GO, no matter what our own instincts are about the timing.

One of my friends in another breed who has produced many litters from frozen semen sums it up best .... "First you need the budget for a large phone bill, second an understanding and very talented veterinarian, and third - nerves of steel.!"

The old rule for using frozen semen used to be only to select a proven dam with a "normal" well documented ovulation pattern and to expect half an average litter size. Nowadays many maiden bitches in many breeds are being successfully inseminated and prod ucing breed-typical sized litters, as was the case with our 1993 insemination which produced nine puppies.

In addition to the three litters since 1992 we now have one from American semen stored for only a few months. We have also had three FAILURES. One of the sires subsequently produced the 1995 puppy from a different dam. The other has been used twice, on tw o different bitches, without success, yet his semen on post thaw examination is of higher quality than that of the others with whom we have succeeded. Both the bitches this semen was used on were already proven dams so there is no obvious reason why to da te his semen has failed to produce puppies.

Although there are many failures, the success rate is increasing all the time. Nowadays our vet is averaging an 80% success rate, mostly by surgical insemination. In 1979, the average was 40%.

Newer techniques are being introduced and perfected. Numerous progesterone levels via blood tests, or daily vaginal smears are taken so that the most accurate assessment can be made as to when to inseminate. Modern day "packaging" of frozen semen is vast ly advanced, in the way of diluents and the provision of buffering solutions to alleviate shock and travel damage.Used only in extreme circumstances in surgical inseminations because the increased volume of fluid may be detrimental, a "waker upper" post t haw buffer is now sometimes used to activate sperm prior to insemination by other methods, for example with fibre optic trans cervical insemintion.

All this progress makes the fact that we obtained puppies in 1976 even more important, because we were all - breeders and veterinarians alike - "flying by the seat of our pants".

It would be interesting - but extremely difficult for the many obvious reasons - for someone to do the statistics on how many natural matings fail as well, but as thawed semen lives for less than one day - some research indicates in fact only a twelve hou r lifespan - and fresh semen lives for three to five days or sometimes more, it is obvious that the timing with frozen semen is extremely crucial.

The most crucial factor is the timing of the individual bitch's ovulation pattern and this in an area in which great advances have been made in recent years.This varies from bitch to bitch and can also vary from season to season in the individual, althoug h another Australian breeder who has been enormously successful with using frozen semen told me that on one occasion he made a last minute decision and simply went by the timing for the bitch's previous A.I. litter, getting a satisfactory number of puppie s without doing any further progesterone levels.

Our vet commences blood tests around the third day and continues every forty eight hours with two, three, four or even more tests depending on how the progesterone levels are developing. Other vets do daily tests using different procedues.

The canine is the only species in which the eggs divide and multiply after ovulation, and therefore surgical inseminations are usually performed several days LATER than when a bitch first indicates she is prepared to stand for a dog, in order to maximise the number of eggs available for fertilisation. Thawed sperm used for surgical implant do not have to undergo the final development stage known as capacitation, which is required of fresh sperm which have to swim to the ova over a period of time. Not all bitches multiply their ova all at the one time, and it is those bitches who do this sequentially for whom it is the most difficult to accurately predict the optimum time for insemination.

Because with surgical insemination the semen in implanted directly, puppies are usually born earlier than 63 days, ours have all been around the 57/58th day, and fully viable.

Many bitches whelp completely naturally but for some reason, it seems that many who have been inseminated with frozen semen do not, so be prepared that a caesarean may be necessary. This is important and breeders need to watch their bitches very closely, especially if only one or two pups have been conceived, as signs of labour are then always very slight, and often a caesar is necessary in this situation even from a natural mating, because labour is triggered by the first pup lining up to be born, with t he others queuing up behind, and with a single pup the bitch very often just does not get the message to go into labour.

Do NOT expect a text book 63 day gestation period if you have used frozen semen, especially by surgical insemination. We exported semen to England a few years ago and the skilled inseminating veterinarian achieved a succesful pregnancy. Tragically however nine puppies were born dead, because the owner had moved from the area and had to consult a local veterinarian who was not experienced with frozen semen. He would not accept this information of a shorter but fully viable gestation period when it was conv eyed to him, and insisted on waiting out not only the average 63 days, but actually let the poor bitch go to her 67th day before deciding to do a caesar as she showed no sign of going into labour. Decomposition of the puppies was of course well advanced, the bitch was by then highly toxic, and has never conceived since despite several natural matings.

We ultrasound to confirm the pregnancy and our veterinarian checks our bitches about the 55th day and x-rays if he deems it necessary, so as to be certain how many puppies to expect. He then decides when they should whelp, based on the technical information gained from the progesterone level tests formerly conducted and of course the date of the insemination. So far he has 100% success rate with his projected whelping date.

With the 1992 and 1995 single pups, we caesared the bitches on the day he predicted. I found this difficult, because they showed no sign of going into labour and in all other respects were outstandingly healthy. My instinct was to wait, as one does with a natural mating until full term as we understand it, 63 days, or at least 59 when many of our own bitches whelo to natural matings. On both occasions, the placenta had already detached and any delay would have almost certainly resulted in the death of the se irreplaceable puppies and been potentially life threatening also for the bitch as she would no doubt have become toxic.

I thought my vet was God the first time, on the second occasion he admitted he was only human and had followed the scientific procedure that has been developed as a result of the increasing use and knowledge of frozen semen. Nevertheless, talking to other people who have lost puppies in similar circumstances, I feel he still does have an almost infallible instinct where this timing is concerned. As I said, you need a very talented veterinarian and to have nerves of steel ....

SEMEN COLLECTION

In order to get a sufficient quantity, i.e. enough semen to make the financial outlay worthwhile, two ejaculates - the correct terminolgy for semen collection - are recommended, our vet prefers 24 hours apart. Sometimes a dog will not be at all co-operati ve on the first attempt, and to be fair it is quite an intimidating experience for him to be in a confined space such as a veterinarian's office, have a "teaser" bitch in season introduced which is quite often of a totally different shape, size and person ality from those of his own breed, and then be expected to allow the veterinarian to handle him with plastic gloves, glass beakers etc. Proven dogs usually manage to sort this all out, and more often than not the second collection is superior to the first.

Personally, we will never collect from unproven dogs as we feel that using them artificially before they have had a natural mating is not a good thing, for either the dog himself or the breed in the long term. We are firm believers in insisting that our f uture studs are capable of managing bitches under natural mating conditions, good and bad, easy and difficult. After all, this is how the majority of their stud work will take place and it also ensures that they thoroughly enjoy their destiny as they shou ld.

I must say I am quite amazed by the number of breeders who don't bother to let their males mate naturally and just automatically do their matings by collecting fresh sperm and artificially inseminating their bitches, but I guess that is another matter as our topic, relates to working with frozen semen.

Sometimes a dog will burst a blood vessel and that semen must be discarded as it is contaminated and useless. This dog may perform better on a second collection and then perhaps the veterinarian will ask for a third collection. All this of course adds to the time and expense that you must be prepared for if you are going to take advantage of AI by frozen semen.

Semen is examined before freezing for motility, sperm density and sperm morphology. If all is in order, then a constant sperm density is achieved by use of a diluent. This is a liquid used to increase the volume of the sperm rich fraction, while protectin g and allowing reactivation on thawing. The earliest formulation for a diluent was known as Anderson's, and it contained glycerol for anti freeze protection, fructose for nutrition, citric acid, and other chemicals as it is essential to maintain a constan t acid-alkali balance, egg yolk, distilled water and antibiotics which were mainly penicillin and streptomycin.

Many advances have been made in this crucial area, and diluents used by the commercial organisations such as the well known ICG - International Canine Genetics - are subject to great secrecy and all the veterinarians belonging to this organisation use the same diluent, uniformity obviously being of considerable advantage when they handle shipments between each other.

The diluted semen is then chilled in a refrigerator for a few hours before being cooled down under controlled conditions to -196 degrees Centigrade.

Our veterinarian likes to send or receive an average of twenty straws, which gives a client sufficient for five surgical implants, four straws being the usual, although some successful pregnancies have resulted from using only three straws where the conce ntration and quality are very high.

For surgical inseminations, using more than four straws at one time is not recommended, because although it would initially seem as though you are increasing your chances, in fact you are decreasing them, because in addition to the semen fraction, each st raw also contains diluent as we have already pointed out, and this overall increase in volumetric liquid would flood the reproductive tract and lessen the chances of conception.

From here the topic becomes extremely technical and is basically out of the breeder's hands and totally in those of the veterinary profession. Apparently different freezing facilities may use different diluents and the concentration per straw can vary, n ot only from sire to sire but also within a single collection. When the semen and diluent are mixed, the concentration can also vary from straw to straw. Apparently there are also straws of differing diameters. Semen is also now being pelletised and packe d in rods as opposed to straws, but I am afraid I have no knowledge of this method.

The most common practice seems to be to freeze at 100 million sperm per millitre, and our vet normally uses four straws per surgical insemination, ie 400 million sperm. He was most concerned that my recently arrived American semen had been greatly over di luted. We received 49 straws but the four that were thawed for the insemination each contained lower than expected sperm content. Fortunately the bitch still managed to conceive one puppy. Now we have just used semen from that same batch on another bitch, and the concentration in these four straws was better than in the first four, although still far from optimal..

Post thaw motility of the sperm provides the critical evaluation of succesful freezing. It may vary between 20% and 90%, with the average being 50% to 70% - any less than this greatly decreases the chance of success and as a general rule semen with lower than 50% motility should not be shipped, or at the very least prior notice given to the client so that they can decide if they wish to take the chance. A straw is thawed at the freezing facility and when semen is shipped it is accompanied by certification as to its health and motility.

Immediately prior to insemination another straw is checked under a microscope for purposes of comparison.

I spent some most interesting time a few weeks ago in Italy with Ludowica Salamon, a young veterinarian having great success with frozen semen and keeping abreast of the comparatively new fibre optic technique for insemination, which I understand is also available in New Zealand.Our vet is doing the same, and will introduce it once his current success rate with surgical insemination is consistently matched by fibre-optics.

You may elect to do your insemination by pipette as opposed to a surgical implant. In this case the preferred method is trans cervical, whereby particular catheters are used by which the cervix can be crossed in order to place the sperm in the womb rather than simply into the vagina.When the bitch is inseminated only into the vagina via a pipette, the success rate is about 52%, whereas when the sperm are lodged in the uterus, the percentage rises and your chances are therefore improved. On an average, you r veterinarian will then use 12 to 18 straws. Two or three inseminations are normally carried out, every 24 hours, at a rate of 4 to 6 straws.

With our currently preferred method, that of surgical insemination, the bitch is lightly anaesthetised for a few minutes whilst a small mid-line incision is made, the uterus is lifted out and the semen injected centrally into both horns and then gently ma ssaged manually into the fallopian tubes and ovaries.As always with anaesthetics given to sighthounds, care must always be taken and our vet uses gas. The bitch wakes up soon after and you take her home to sweat out the next 21 days - you that is, not the bitch who could of course not care less that she might be about to make history - until it is time to see if she has conceived.

Sometimes veterinarians do one insemination by surgical implant and use additional semen for a second insemination by the conventional pipette method. This would be logical where - for example - you either have vast amounts of semen at your disposal and c an afford to be extravagant, or alternatively you have only managed to get a total of six or seven straws. With the latter, you would then have to decide a) to try it all at once but by employing both methods on a single individual on different days, or b) use up four by surgical implant on one bitch and three on another at a later date. We are in exactly this position with the remaining semen from one of our precious long-deceased imports.

NOW LET US LOOK AT SOME ETHICS

So far working with frozen semen has been very much on a breeder to breeder basis and not really conducted at a commercial level. Arrangements have been rather casual, and while a few grumbles have been heard, nothing dramatic other than reports of defect ive semen being shipped seems to have occurred. However, as we have said, times may be changing.

Firstly the question of how much semen a buyer is entitled to needs to be addressed.

We have said an average shipment of 20 straws is the accepted practice, so what then constitutes a "fair thing" when you are asked to supply semen or decide to import some for your bloodlines?

In practice, it seems that most breeders - owners of stud dogs and of potential dams - who are prepared to go through the time, money and hassle of AI by frozen semen do all this because they want the breed to benefit from the dissemination and distributi on of genetic material which would not be available otherwise.

Most stud dog owners seem to be happy to agree that provided the actual "up front" expenses of collection, freight, Governing Canine Body registrations etc are paid in advance, they are prepared to allow unlimited access to whatever quantity is shipped.

Some consider these expenses as being equal to a stud fee, others ask for one stud fee to be paid at the same time the initial costs. Some owners request that further stud fees be paid each time a successful pregnancy results. Just be sure that you both a gree to whichever is to be the arrangement, and preferably do this in writing so that both parties are protected.

Normally, when you pay a stud fee for a natural mating, most breeders will give you two matings TO THE SAME BITCH IN THE SAME SEASON, FOR THAT FEE. Many will offer a "free return" should the bitch fail to conceive, but of course, they are under no obligat ion to do so. Extended to frozen semen, this would equate to two attempts with one bitch (i.e. should she miss the first time) and the option to then try another bitch if unsuccessful. In practice, few people would try a second time with a bitch who has f ailed to conceive to frozen semen as we have alredy told you that some bitches simply fail for no apparent reason. Most breeders would prefer to select another bitch for the "free return".

As opposed to the expenses with natural matings, the importer of frozen semen has already paid a considerable amount for up-front costs, on average this will be the equivalent of a second stud fee, and each surgical insemination will costs around the same amount again So it really comes down to whether the stud dog owner is supplying semen in order to earn money or because they genuinely wish to see their dog used somewhere else in the world, and as we have said, provided of course that they are not out of pocket themselves, most are happy for the shipment of semen to be used as their client deems necessary.

Having dealt with potential usage, we need to look at quantity. With good management when a dog is presented for a collection and he is prolific, he will provide more than enough semen for use in a frozen semen programme. The owner of the sire may then de cide to freeze any additional semen for their own future use.

The amount of straws to be sent, depending of course on the quality and volume of the collection, is also a matter which should be clearly agreed to between the parties before the semen is shipped. Very often - especially if both owners are really keen to see a successful result - they agree that the entire collection will be sent so that plenty of semen is available at its destination to cover all the varied contingencies which can result in failure to get puppies despite every one's best efforts. For i nstance, if a live litter is born but does not reach maturity, there needs to be agreement as to whether that constitutes a litter as such.

To avoid any later unpleasantness, the sensible thing as we have said, is to work out in advance how many litters each considers the semen will entitle the importer of the semen to have, and at what stage - if any - a further stud fee will be payable. Obv iously the number of litters possible will vary according to the amount of straws used in the method chosen for insemination. Sometimes clients feel that if they pay the stud fee, then they automatically expect every drop of semen collected on that occasion to be sent to them. Seems logical, but think it through.This is fine in principle, but in fact with an average collection o f twenty straws do not forget there is enough for FIVE inseminations, so if the importer has the potential for so many litters they are in fact doing very well. Often, also of course, it is simply not possible for many kennels to take advantage of multip le inseminations from the same sire, so twenty straws are far more than will be used over most breeding programmes. It is not really justifiable to take every drop from the collection unless the importer has enough bitches to use it on.

If there is a great deal of semen, more than most breeders can logically use, why not come to an agreement that the sires'owner may freeze the excess for his own purposes later on? After all, when the collection is taking place, it is fairly illogical for the owner not to start storing some for themselves, as a subsequent trip to the veterinarian will of course attract a whole new lot of costs.

The question is often asked WHO OWNS THE SEMEN ONCE IT CHANGES HANDS?

In our country, imported semen becomes the property of the importer. Service certificates are signed by the inseminating veterinarian. At this stage any arrangements that may have been made between the shipper and the importer such as stud fees payable ea ch time semen is used etc. are considered private and a matter of trust. The only way this can be policed by the Controlling Body would be to insist that the owner of the dog signs each certificate as well. We did have a very unpleasant case in Australia where this had not been clearly spelt out and the owner of the stud dog belatedly decided they wanted a stud fee for each mating.

Probably as the use of frozen semen is increasing so rapidly, a world wide policy should be agreed upon between all the Canine Governing Bodies and a simple document developed where the shipper can select a box granting permission for the agreed number of inseminations, beyond which a stud fee musts be paid. If this were then signed by the importer, there should be no further problems and it could be lodged at the same time as the donor dog is re-registered in the country to which the semen has gone .

Something else which needs to be seriously considered is whether or not the stud dog owner is agreeable to semen being on sold by the original purchaser. Personally I would not agree to this under any circumstances unless advised of the potential destinat ion and given time to consider the suitability of the pedigrees in that kennel. However it has happened on numerous occasions, presumably with the dog's owner at least paid the courtesy of being asked for their permission. It is not really a matter for t he Controlling Body, because if they have ruled that the importer owns the semen, then presumably in their eyes he has the right to dispose of it as he wishes. When I export semen, I formerly advise my client in writing that I do not agree to it being on sold without consultation. Legally I cannot stop them, morally I hope they feel obligated to meet my requirements.

Another area where problems could occur is when semen is collected by the current owner before selling or giving a dog to another person.Obviously ethics dictate that the new owner be informed of this. In our case, when we sell a male from whom we have co llected, we would never make that semen available to another person DURING THE LIFETIME OF THAT DOG, although obviously we would expect to sell any progeny we may produce from the semen, without opposition from the dog's current owners. Any stud enquiries would be re-directed to the new owner, which we believe is the only correct way to behave in this situation. Should the dog be deceased but semen from our collection still available, then it would depend on whether or not we could spare any of it. If we could, we would expect the stud fee to be payable to ourselves, and not to the person to whom the dog had been sold, as he was our property at the time the semen was collected.

Now what if you GIVE - as opposed to sell - a dog to another person and they then decide to collect and store semen and then the dog dies? Who owns that semen, you because you did not charge for the dog so technically he was still YOURS, or do they, who h ave had the foresight to collect?

My feeling is that if they paid for the collection, they own the semen, but you can see this could be a mine field and should be taken into consideration.

Which raises the question of whether breeders should be allowed to use frozen semen from deceased dogs, or should they not? Obviously we personally believe that far from preventing this, Controlling Bodies should in fact be encouraging it, especially with relation to hereditable disease control.

Of course people say "How can they prove those puppies are by that dog now that he is dead?" Our answer is " Other than by the very expensive use of a DNA test, how can you prove ANY puppies are by ANY sire to whom they are registered?"

Unless you are personally acquainted with the sire himself and were actually present when he mated the bitch in question and unless that bitch was under your personal observation for the entire time of her season so that you can swear there is no possibil ity of a mis-mating having taken place, how CAN you prove it?

The same could be said of puppies born from bitches already in whelp when imported to another country.

The keyword in breeding livestock is TRUST. If a breeder is ethical, then they will apply the same strict standards of honesty to their frozen semen programme as they do to puppies they produce from natural matings. If you would buy one from a breeder bec ause you like the type they produce and the pedigree interests you, then you must be prepared to trust them equally as much when they announce puppies from frozen semen.

Don't do what one other breeder did recently - he predicted puppies of a certain colour to be born from frozen semen - in someone else's litter of course! - and they were not. However two other colours, which he had not predicted, were. He carelessly com mented "But that dog never produced black and tan or black masked gold when he was alive". To his embarrassment it was promptly pointed out that in fact two of the best known of the many champions by the sire in question were exactly those colours. The bl ack and tan had become an important brood bitch for a well known kennel and her lines are still prominent today. The black masked gold was a major winner and took Best In Show at a specialty under a very famous overseas breeder, one of the few to officiat e in Australia in those days. He was a very dominant producer and also features several times in the pedigree this person was doubting.

COSTS

As far as cost was concerned with our 1976 litter, it turned out to be far more expensive than we had first estimated. It was all experimental and very time consuming for all concerned and we gratefully recompensed those involved.

Costs nowadays have decreased considerably, because there is a system in place, many tasks are automated, more owners share space in shipping canisters etc. In Australia, we need to allow around $1000 US in expenses to obtain a successful pregnancy and th at is not counting whatever stud fee arrangements we may be able to make with the owner of our chosen sire.

TO SUM UP

So you can see that importing semen is an exercise which needs to be weighed up most carefully. Of course it is possible to bring in multiple doses of semen in the one canister and thus inseminate a variety of bitches, but each insemination is costly and it is far easier and much quicker to incorporate a new bloodline by importing a live stud dog.

Responsible breeders will take years to incorporate, blend and develop the new genes with their existing ones - the object of using frozen semen, as it is with natural matings, should be to consolidate the known benefits of one's own lines while at the sa me time introducing perceived benefits from someone else's. As with natural matings - sometimes you get what you want and sometimes you don't.

Overall of course you would expect to get more mileage by importing a male whom you could use on numerous occasions over a variety of bitches for the remainder of his fertile life and thus establish a widely based gene pool. Against this, one needs to wei gh the enormous advantages we have already discussed of using frozen semen from a sire you have especially selected.

As breeders who have both imported numerous live dogs from several countries in a number of breeds, and been successful with the importation of frozen semen plus the use of long stored frozen semen from our own stud dogs, we can only comment that both met hods have been invaluable to our breeding programme and have contributed to the successful development of our gene pool.

ADVANTAGES

Matings that could never occur can now take place

Overcoming geographic isolation and avoiding the disadvantages of quarantine

Long-term storage
a) for intensive line breeding to famous dogs who may by then be deceased
b) for potential use of valuable genetic material which at the time of collection is too close to your current bitches

As an extremely valuable tool in those breeds afflicted with hereditary diseases

DISADVANTAGES

Danger of a new but basically incorrect or untypical "look" becoming too easy to source

Risk of entrenching hereditary diseases more widely geographically and more rapidly throughout the gene pool

Temptation for insufficiently researched breeding to well advertised top winners with very limited line-breeding becoming accepted practice, Consistent out crossing must inevitably lead to a weakening of predictablility for type

HISTORY

1976
First frozen semen puppies registered with a Canine Controlling Body, Australia

Late 1980s
Success being achieved world wide. New techniques and management practices developed

First success with long stored frozen semen,
1992 With 16 year old semen
1993 With 17 year old semen
1995 With 19 year old semen

PROCEDURE for obtaining semen

Find an extremely experienced veterinarian working in the field of frozen semen before you even THINK about importing any

Select a stud dog and a recipient bitch, making your plans at least 6 months in advance

Check with your Controlling Body and Government Department for all requisite paperwork. MAKE SURE YOU GET THIS RIGHT

Forward details to owner of stud dog SO THAT THEY CAN MAKE SURE THEY ALSO GET IT RIGHT

Make your arrangements with the owner concerning stud fees, amount of semen, any restrictions for on selling, and so forth, all preferably in writing and signed by both parties

PROCEDURE at time of insemination

Advise your vet the first day your bitch comes into season so that he can plan accordingly

Decide if you want a surgical insemination or trans cervical by pipette and catheter

Unless the pregnancy is very obvious, ultrasound at around four weeks

Again, unles the pregnancy is very obvious, be prepared for an x ray in the eighth week

Do not automatically expect a full term of 59 to 63 days, and watch your bitch extremely closely for any signs of labour from the 55th day

Be prepared for a caesarean if your vet deems it necessary

ETHICS

How much semen does your financial outlay entitle you to?

Who owns the semen after collection and shipping?

Should AI from deceased dogs be permitted?

Wendye Slatyer July 97
Copyright(c) 1997

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