continued AND AWAY WE GO...
Collecting and
Preparing Semen for Boyage
Republished
with permission from Horse Sport
Sperm Counting Device
After the microscope and the incubator, probably the next most important
piece of equipment is some form of counting device to establish the
concentration in millions per mL (abbreviated as: number x 106/mL).
There are commercially available units, the "king" of which
is the Densimeter. The Densimeter not only establishes the concentration
of the sample, but will also calculate the required volume to establish
the correct insemination dose. Other counting devices, not quite as
fancy as the Densimeter, are the "SpermaCue" and the
"HRI Sperm Counter". These devices all operate on the
principal of a spectrophotometer, which is a machine that measures the
light transference through some sort of medium. In fact, you can use
a spectrophotometer to do the job, the "Spec 20" model is
the standard of the industry, and this is what we use at Atlantic A.I.
Services, as we are able to calibrate it for different species (we also
freeze dog semen). If you do elect to go with a spectrophotometer, you
will have to get it calibrated for equine semen and Colorado State University
will do that at a minimal cost, with little delay.
If all of the above units are outside your pocketbook range when starting
(they are priced at about $1,500 and up), then you will have to resort
to the somewhat more time-consuming, albeit actually more accurate,
hemacytometer (various manufacturers, cost is around $100). In
fact, once you have become familiar with the hemacytometer, and have
had some practice, you should be able to establish concentration levels
within 10 minutes.
Unless you are using the Densimeter, and even then on occasions, a calculator
will save a lot of time and mathematical effort when calculating insemination
doses and extender rates.
Syringes or pipettes will be required for handling the semen, and for
getting the correct amount for calculating concentration. If you are
using syringes, they should be of the all-plastic, non-spermicidal type
and not the normal rubber-plungered type.
Semen Extenders
There are a variety of semen extenders available commercially, the majority
of which are based on Dr. Bob Kenney's tried-and-true non-fat-dried-milk-solid
(NFDMS) "Kenney" formula. The only variable in the
majority of these is the antibiotic that is added to it (if in fact
there is one). It is advisable when first collecting your stallion each
season, to split his ejaculate and store it in several different extenders
(i.e. different antibiotics) to establish which he seems to be most
compatible with. There is usually a little variation, although sometimes
it is negligible. The two antibiotics that I favour the most are Amikacin
Sulfate (which is in the "EZ Mixin' CST" extender from
ARS), and Ticarcillin (although this has now been replaced by "Timentin"
which is a mixture of Ticarcillin and Clavulinic Acid. This antibiotic
is not as far as I am aware, available in a commercially prepared extender).
I tend to stay well away from using Polymixin B. Sulfate in any but
on-farm inseminations, as I have found it seems to be more detrimental
to longevity that others. Another commonly used antibiotic is Gentamycin,
although I have run across some that have the same concerns about the
use of this as I do the Polymixin B. Sulfate.
Essentially, for the beginner, it is best to go with one of the many
commercially available semen extenders, rather than attempting to mix
your own.
Semen Transport Containers
Without a doubt the best of these is the "Equitainer"
(Hamilton-Thorne Research). It is also the most expensive initially,
but will last for many, many years, and so become by far the cheapest
in the long run. (I know of several farms that are still using the old
blue-bucket type that was the original prototype developed almost 20
years ago!). The Equitainer is the most reliable as far as insulation
properties against heat or cold is concerned, and its tough plastic
shell is immensely durable and will handle the rough handling of courier
companies and airline baggage handlers.
There are several disposable containers available for use with
transported semen as well, such as the "Bioflite";
"ExpectaFoal"; and "Equine Express II".
These are all constructed using a cardboard box and styrofoam insulation.
The manufacturers recommend they be used no more than fivetimes, and
fewer if damage is evident. The trouble with these systems is that they
tend to have poorer insulation qualities to start with, and then if
they are subject to rough handling in transit, they can develop microcracks
which may not even be visible. These microcracks can allow the semen
temperature to drastically alter.
The price certainly is a bonus initially (usually around $35-40, as
opposed to Equitainer's $300), and some farms simply add the cost into
the stud fee, and don't bother about trying to get the mare owner to
return them (getting shipment containers returned can be a nuisance
occasionally). The Equine Express II allows for the packaging of the
semen in the insemination syringe, which may be attractive to some,
but means you will have the added expense of providing the all plastic
non-spermicidal syringes. If using one of the other systems, you will
require baby bottle liners and elastic bands to package the semen in.
A permanent marker is also required to write identification on the outside
of the bag or the syringe.
Outside Equipment
The first consideration for "outside" equipment is whether
you are going to use a breeding phantom (phantom mare or dummy),
or a live jump mare. If you are going to use a phantom mare, then obviously
that will have to be bought or constructed. I have built many very serviceable
breeding phantoms using sections of telephone pole.
Regardless of the type of "mare" you are using, the stallion
should have his penis washed off prior to a collection. For this you
will require a source of warm water, a bucket, a sterile liner for the
bucket (disposable plastic bags will usually suffice), and a sterile
plastic cup.
It is quite possible you will need somewhere to "tease" the
stallion, to encourage him to "drop" for the washing process,
and then to gain an erection prior to collection. Depending on the stallion's
temperament, this teasing can be achieved over a stall door or through
stall bars, or it may be that a proper teasing rail is preferred - especially
with rowdier stallions that may become too used to teasing over a stall
door, and attempt it each time they pass one. If using a breeding phantom,
some stallions may require the tease mare to be in a close proximity
to the phantom, as they may become distracted if the distance is too
great, and loose their erection.
The above is a list of basic equipment that the person collecting semen
for on-farm or cooled-transported use will need. There are many other
pieces of equipment that can make the process more efficient, easier,
and a whole lot more expensive.
For example: centrifuges, a slide warmer, a refrigerator, a water bath
(if you have an incubator for regular use), various sizes of pipettors,
and if you have money to spare, the holy of holies: CASA, which is "Computer
Assisted Semen Analysis" - priced at $25,000 the last time I checked!
In addition to his A.I. business, Mottershead also performs stallion
behaviour modification, including retraining of stallions for breeding
and initial training for semen collection, and
conducts research into several aspects of equine reproduction and emphasizes
assisting breeder knowledge through
his website, www.equine-reproduction.com, and by co-moderating
the largest non-restricted equine
reproduction e-mail list on the Internet, EquineRepro@egroups.com
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