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Excellent Artificial Insemination Article and Information Courtesy of Katriina

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The artificial insemination of dogs is nowadays a international hobby. There is a open transfer of information between countries due to kennel associations, magazines, the internet, and international judge exchange in dog shows. All this has increased the interest in international exchange of dog sperm, and in addition, posed new challenges for veterinarians in artificial insemination and the storing of sperm. Especially in countries that have strict quarantine regulations, the possibilities of receiving new genetic material from the world’s best dogs, has turned a new leaf in the history of the breeding of dogs. A veterinarian can also be of help in such cases where the female and male dog live far away from each other. Sending the sperm and using artificial insemination can spare the owner from a long, expensive and tedious journey. In the future, new efficient methods in freezing sperm will become essential for the breeder Artificial insemination allows using the best possible male dog irrespective of distance, and time, since frozen sperm can be utilised decades after the death of the male dog. Especially, when widening the narrow basis of breeding of rare breeds, and in fighting hereditary deficiencies, artificial insemination will prove out to be a significant method.
Methods of Artificial Insemination:

Artificial Insemination with Fresh Sperm
Artificial Insemination with Transported Sperm
Artificial Insemination with Frozen Sperm

1. Artificial Insemination with Fresh Sperm
Both the male and female dog are present at the artificial insemination. Acceptable reasons for artificial insemination are, among others, the following:

A medical reason; back or foot defect, which prevent copulation
The female dog has a tight residue of the hymen (this problem usually disappears after giving birth to first litter).
If there is a possibility that the male dog has an infection which may pass on to the female dog
If the male is young and over-active, and the copulation does not succeed
If the male lives in the same household with a dominating female and does not want to copulate
If the female does not accept the male (this is common with dogs which live in the same household)

In practice the most common situation is that the female does not allow copulation, or the male is not interested in it, but the time is suitable according to the owner. Research has indicated that the behaviour of the female may vary significantly, even with the same female between different estruses, not to mention, between different females. The best time for copulation is generally the 12th to the 14th day from the start of the heat, but the variation is great: there are females for which the best time is already over on the 7th day and others for which the best time is on the 25th day. All days between these time limits are possible and normal.
Because of such great variation the time should always be verified with a hormone analysis, especially if the copulation is unsuccessful or the time is otherwise uncertain. When planning artificial insemination the appropriate time is always defined with a hormone evaluation. If the sample is taken in Veter in the morning before 10.00 o’clock, the result is received already at 02.00 o’clock in the afternoon the same day. If the sample is taken outside the Tampere region, the result will be received from the animal laboratory Vetlab after 02.00 pm the next day.
If the heat of the female seems to be passing, the artificial insemination can be carried out, but the situation must always be assured with a simultaneous progesterone sample. When the result of the progesterone level is received, we will know if the time for the artificial insemination was right or when the new insemination should be carried out.
When the time of the insemination is near, the progesterone analysis can be utilised to define approximately 2-3 days in advance when the time is appropriate for the procedure. The quick progesterone analyses are not useful, since their accuracy varies to such an extent that there may be a difference of one to three days in the estimation. They can be used to define the suitable time for the actual progesterone analysis.

The Actual Artificial Insemination
The artificial insemination can sometimes be performed into the vagina, but better results in pregnancy are achieved when the insemination is performed directly into the uterus.
Artificial insemination into the vagina is successful if the time is early enough; for the female dog the cervix of the uterus closes when the progesterone level reaches 10 – 20 ng/ml, and after which copulation or artificial insemination into the vagina is unsuccessful (the spermatozoa cannot enter the uterus and thereon to the oviduct to fertilise the ovum).
Artificial insemination into the uterus is usually performed only at the end of the oestrus, when using fresh sperm the level of progesterone is 10 – 15 ng/ml, and with frozen sperm around 15 – 20 ng/ml. The closing of the cervix of the uterus does not prevent artificial insemination into the uterus with a catheter.
The artificial insemination begins by collecting the sperm from the male when it becomes interested in the female. The number and movement of the spermatozoa is analysed after the sperm is collected, and if the sperm is of good quality the artificial insemination into the uterus is performed on the lightly sedated female. After the operation the female is given a stimulus and within a quarter of an hour it is ready to be taken home.
The gestation results depend entirely on how precisely the moment of insemination has been defined, and whether the insemination is done into the uterus. If the insemination is performed into the uterus on the most suitable day according to a hormone definition, the possibility of pregnancy is better than with normal copulation (the closing of the cervix of the uterus does not prevent the pregnancy, and sperm of not so high a quality works).

2. Artificial Insemination with Transported Sperm
Artificial insemination with transported sperm refers to a situation where the male dog lives abroad, usually somewhere in Europe or lives within a long distance in Finland. Similarly, diluted fresh sperm is sent all over Europe from Finland. The sperm can be diluted and cooled to a temperature of few centigrade Celsius to preserve it for 1 – 2 days.
Generally, the sperm is collected in the morning in the sending country. After dilution and cooling, the sperm is transported directly to the airport. To ensure that the sperm shipment arrives at the destination, it is best to choose a direct flight to Helsinki. If there are intermediate landings, the shipment may arrive even a day late. The insemination is carried out on the evening of the same day in the receiving country. Another possibility is collecting the sperm in the evening, and sending it on an overnight flight to the receiver for the insemination.
The owner of the female dog, which is to be artificially inseminated, arranges the collection of the sperm and shipment with the owner of the male dog according to the instructions received from the veterinarian who performs the procedure, and collects the sperm shipment from the freight station at the airport and brings it to the location of the insemination.
Nowadays we only use the GLONE kit for sending the sperm. The kit consists of a styrox shipment box, a cooler, and the needed dilution and stimulant fluids, and exact instruction on collecting, diluting and the handling the sperm before the insemination. The information is important, since artificial insemination is still relatively rare and new in many parts of Europe; exact instructions are thus often crucial. We still occasionally receive spoiled fresh sperm shipments.
When the sperm is sent from Finland to another country, the sperm is collected with the help of a rutting female dog, analysed and diluted. When using the kit, the sperm can be packed directly into the sending package with the cooler. When the sperm is packed adequately it gradually cools down to a few degrees centigrade. The sperm collection form and other information are packed inside the shipment. The owner takes the package directly to the airport and sends it to the receiver.
The time of the artificial insemination is always defined with the progesterone analysis; the insemination is performed when the progesterone level is 10 – 15 ng/ml (in other parts of Europe the scale is: 10- 15 ng/ml = 30 – 45 mmol/l). If the insemination is performed into the uterus, the level of progesterone may be up to 20 ng/ml (60 mmol/l). It is preferable to perform the insemination into the uterus, because of higher pregnancy rates.

3. Artificial Insemination with Frozen Sperm
Frozen sperm is always used when the transfer distance is long, or when the sperm of a good male dog is collected in advance for future use. Unfortunately, many owners realise this latter possibility only when the male dog is relatively old. The number and the quality of the spermatozoa start to decline at the age of 6-8 years among most male dogs. Freezing and defrosting deteriorate the movement and quality of the spermatozoa the more, the lower quality the sperm is when collected. Sperm for 3- 6 artificial inseminations can be obtained during one collection from a relatively young male, which weighs 30kg. The obtained number of portions of sperm needed for an artificial insemination is affected by the dog’s size, age, the quality of the sperm (inflammations may deteriorate the sperm), recent copulations, the level of arousal of the male, etc.

4. Sending Frozen Sperm

When the owner of the female dog wants the artificial insemination to be performed with the sperm of a foreign male dog, the owner finds the suitable donor. When the donor male is found, the owner of the female dog asks the owner of the male dog to have the desired amount of sperm frozen in the local sperm bank. The freezer is informed on how many portions of sperm are needed. After which the freezer examines the quality of the sperm and defines the needed amount for the needed portions to be sent. The freezing has to be performed already in advance. It is possible to send only the needed amount of the frozen sperm, and the rest can be kept stored in the sperm bank to be used later either in the homeland or somewhere else in the world.
	Dog sperm can be imported to Finland without any importing licences or antibody definitions. This is sometimes hard to believe in other countries! The sperm bank always takes care of sending the frozen sperm. Forwarding agents are used in shipping between two countries. The sperm is sent in a liquid nitrogen container, which is rented from the sperm bank. During the shipment the sperm has to be in the container in the temperature of minus 120 - 196 centigrade Celsius. All forwarding agents do not transport nitrogen containers. Currently, we have an agreement with FedEx (Federal Express) on the customs clearance and transport of nitrogen containers to our practice. FedEx is able to handle transportation all over the world.

If the shipment arrives to Finland from outside of the EU by some other means, the customs clearance, collecting from the customs office and returning it to the sender is exceedingly difficult (customs and EU regulations). In practise, the sending sperm bank fills in a FedEx form and makes a Pro Forma bill and a clarification of the way the container will be sent and returned. When the container is filled accordingly with liquid nitrogen, and the diluted frozen sperm in straws is packed into the container, the sperm bank attaches the information on the quality of the sperm to the container and requests the local FedEx to transport it. The forwarding agent takes care of the temporary customs clearance of the container according to regulations and transports it to the correct address in the receiving country. When it has arrived to its destination, the sperm is transferred into Veter’s sperm bank and the attached information on the male dog and the quality of the sperm is stored for later use. The container is immediately returned according to our instructions to the sender via FedEx. The basic rent of the container covers the first week, but after that an amount of 10 euros is charged for each additional day. Sending sperm abroad from Finland is done similarly. Australia is an exception; the sperm can be transported into this country only in a new container or a container sent from Australia for the shipment. In addition, a importing licence (10 pages) has to be acquired from the local authorities and the container must be sealed with a seal by a licensed veterinarian when shipped. Many countries require results from brucella and leptospiroses antibody analyses (e.g. Sweden, Norway and England). Some countries require also the DNA-chart of the male dog before they accept the sperm for use (USA and Canada). DNA-chartings are nowadays also done in Finland or a cell sample can be sent to a DNA laboratory in the USA.

5. Sending Costs
The costs of sending frozen sperm consist of several different factors. I have collected here some estimated example prices of sending sperm to various countries.

The liquid nitrogen for filling the container (Aga) 115 e
The filling in of customs, shipment and other required papers, and the transport of the straws 100 e
The rent for the liquid nitrogen container (+ 10 e per each day after the time of one week) 100 e / week
FedEx transport from Finland to USA 380 e
FedEx transport back (from USA to Finland) 380 e
FedEx transport from Finland to Europe 140 e
FedEx transport back (from Europe to Finland) 140 e
When sending sperm from Finland, the receiver usually pays both the freezing and shipment expenses. The general policy is that the receiver sends us his/her credit card number and the date of validity, so we can charge the abovementioned expenses. The expenses equalling coupling fees are agreed among the owners.

Freezing Sperm

Freezing dog sperm is a long process and it requires the time of half a day. Therefore the freezing time has to be agreed upon early on before hand. Sometimes we have faced situations where the male is about to leave the country, but the owner would still like to have its sperm frozen. In most cases we have been able to arrange this.
A rutting female dog, which the male dog is interested in, is needed when the sperm is collected. Without the female dog, no spermatozoa are acquired, or the number of spermatozoa is significantly smaller, than it would have been with the female. Generally the largest amount of sperm is desired, even so that at the same occasion sperm can be collected again after an hour, and the sperm acquired with the two collections can be frozen together.

In Veter we utilise the GLONE method for freezing; with this method the mobility of the spermatozoa is generally >70% after defrosting. With other methods the mobility rates of defrosted spermatozoa is often about 40 – 50 % and even lower. Similarly, the spermatozoa maintain their mobility for up to 3 days after defrosting in refrigerator temperatures, whereas, with other methods there is virtually no mobility one day after defrosting. These features significantly affect the likeability of pregnancy. Research has shown that the likelihood of inducing pregnancy of the sperm starts to decline rapidly after the mobility decreases below 50%.

When the sperm is collected, the male should be identity marked, either with a microchip or a tattoo. The registration papers are needed for storing the information. There are several male dogs even in Finland that have their sperm collected regularly and sent all over the world when needed.


We only need the following information:

The address where the sperm is sent
Information on the owner of the female dog
How many inseminations the sperm is used for (The amount of the sperm to be sent, is defined according to the quality of the sperm, not the need of the receiver)
When the sperm should arrive at its destination If the receiver pays for the FedEx transport expenses, we need his/her FedEx number, or credit card number, and the date of validity - otherwise the shipment has to be paid in Finland.

According to this information, we pack the correct amount of straws and dilution fluid in the container and send it to the receiver. In Finland, the owner of the sperm only has to supply us with the abovementioned information.

: We have had a sperm bank for dogs in Veter for 15 years. Currently, we have the sperm of 100 males waiting for artificial insemination or shipment. Sperm is conserved unaltered in
–196°C for an infinitely long time.

Freezing Costs
Collecting, analysing and freezing the sperm 420 e
Additional collection of sperm after an hour 100 e

Tariffs for the Storing of Sperm (2003)
One cassette (10 straws), charged annually in January 115 e / year

The sperm of each male is stored in a cassette marked with an individual code. There is space for 10 straws in a cassette. In average, two straws are used for one artificial insemination. One straw of thick, high quality sperm is sufficient (0,5 ml) for one insemination. Correspondingly, 3 – 5 straws of lower quality sperm has to be used. There can only be sperm from one male in one cassette, therefore the storage of one straw costs the same as one full cassette. Sperm which is sent from abroad may also be frozen in pills or mini straws (0,25 ml). These can be stored like ordinary straws, whereas the defrosting of the sperm is done accordingly.

Artificial Insemination with Frozen Sperm

Artificial Insemination with Frozen Sperm
At this point the sperm is in the sperm bank. The veterinarian should be contacted when the female dog’s oestrus starts; now it is possible to estimate the date of the artificial insemination. When the heat progresses, the first progesterone tests are run when male dogs begin to show interest in the female, the menstrual flux is lighter in colour, or after 10 – 12 days from the start if there are no other visible signs of progress. Samples have to be taken until two results that show high levels of progesterone (5 – 20 ng/ml). If the samples are taken outside Tampere region, the local veterinarian marks these results into the admission note, so that we receive this information in Veter.

The owner can ask for results from Veter the next day after 02.00 pm. If the sample is taken in Veter before 10.00 o’clock in the morning, the result is obtained on the same day. If the time of the artificial insemination falls on a weekend, the last sample is usually taken in Veter already on Friday morning to obtain information for the weekend. The artificial insemination is performed on a slightly sedated dog with a catheter into the uterus. At the same time, when defrosted, the quality of the sperm is examined. The rear end of the female dog is held lifted for a while after the procedure, and a stimulant is then given. If clearly indicative progesterone results are received, only one artificial insemination is performed during one oestrus.

Artificial insemination Tariffs (autumn 2003)
Artificial insemination into the uterus with frozen sperm 290 e
A second artificial insemination into the uterus with frozen sperm 200 e
Artificial insemination into the uterus with fresh sperm 220 e

Results in Artificial Insemination
Most of the dogs artificially inseminated in Veter come from Helsinki region. However, dogs come from far away as well, even from Tallinn and Moscow.
We send out and receive sperm shipments weekly. Most of our shipments are sent to USA, but also to South-Africa, Canada, Mexico and various countries in Europe.
In Veter we have performed artificial inseminations with frozen sperm for 15 years. The first dog sperm bank has functioned the same time.
Annually, we perform approximately 30 artificial inseminations with frozen sperm in Veter. The number has been on the increase. According to the Finnish Kennel Association, in the whole of Finland, 60- 70 artificial inseminations are performed annually.
Practice has shown that artificial insemination into the uterus for a dog is such a demanding and challenging procedure, that it requires long-time experience. The results depend on the experience of the veterinarian who performs the procedure. For example, in Sweden and Norway artificial insemination with frozen sperm is only performed in universities. In these countries it is also prohibited to perform artificial insemination with surgery.
The pregnancy rates have varied
• when we first started they were 75 – 80%
• currently they are >80%