Laparoscopic Artificial Insemination, also referred to as LAI, is the minimally invasive surgical method used to deposit semen directly into the lumen or inside of the uterus. This technique was first developed in Australia in the 1980’s with extensive work done by the University of Sydney.
The ewes are given a mild sedation to allow them a more relaxed stress-free procedure. They are then placed into a surgical cradle on their backs. These cradles are designed specifically for the purpose of LAI which allows them to be inverted at a 45-degree angle. This position allows the abdominal contents to be displaced away from the location of the uterus. This aids in the working space and visualization of the reproductive tract. Once the veterinarian locates the uterus a surgical assistant transfers the semen from the water bath into an insemination pipette. The assistant then passes the pipette through the working port of the laparoscope. The veterinarian then deposits the semen into the uterus.
The surgical procedure lasts less than a minute, allowing the animal to be returned to the recovery pen very soon. Animals are administered a long-acting antibiotic, surgical sites are covered with an antiseptic spray prior to being uncradled.
Embryo Transfer or ET is a tool used to increase or maximize the production of genetically superior individuals. Candidates for embryo transfer generally fall into one of the following groups. The first group includes, young to middle-aged highly productive females where multiple offspring per year is desired. The second group includes older or debilitated females who are unable to carry or care for their offspring and yet remain superior producers.
An embryo transfer program can be set up either on farm or in-house at RSG. Both involve significant amounts of time and resources to be successful. All of the same management principles apply to programs on the farm or in-house in order to maximize success. The highest of health standards and production management must be met. For on farm programs, a minimum of ten donors need to be assembled with a minimum of five recipients per donor for programming.
For ET programs at RSG, donors need to be in place at least four weeks prior to the beginning of the flush program. A minimum set of health standards are required, including, no more than 60 days of fleece, trimmed hooves, free of foot rot and scald, and dewormed within a week of arrival. Donors arriving at RSG that do not meet these standards will be subject to rejection from the program or may incur significant charges for bringing the donor up to specifications. If rejected from the program your deposit will not be refunded. All donors must undergo a quarantine period to ensure the health and safety of the others in the program.
Once the synchronization program has begun, it will take approximately 20 days to complete. Donors will be initially synchronized in a similar fashion as LAI program. However, in the days prior to breeding, they will undergo twice daily injections of Folltropin V, a hormone to induce superovulation. The females are then bred with LAI using either fresh or frozen semen. High-quality fresh semen is preferred and allows for the greatest success rate of embryo fertilization. Frozen semen is a viable option; however, multiple inseminations are required in order to ensure viable semen at the time of fertilization. Frozen semen will be evaluated prior to insemination and may be rejected if minimum quality standards are not met. Clients must have a backup plan in place in the event that the semen, fresh or frozen, is evaluated and the quality is not sufficient for fertilization.
Recipients will be synchronized at the same time as the donors to ensure the uterine environment is ideal for embryo implantation. Teaser rams are used in conjunction with the synchronization hormones to add another measure of response.
The embryo donors will undergo the embryo collection, or flush procedure, six days following breeding. The embryo recovery is a surgical collection procedure where the donor will be placed under general anesthesia, positioned in dorsal recumbency, and surgically prepped. A trained technician serves as our anesthetist to ensure the highest quality of surgical anesthesia possible. The surgical collection is then performed by Dr.Tad Thompson, DVM. Once the collection is completed, Dr. Thompson will place the uterus back into the abdomen and use a standard closure technique to complete the surgery. Many steps are taken to ensure proper sterile surgical technique and appropriate surgical practices in order to reduce the risks of complications such as infections, adhesions, or dehiscence. However, as with any surgical procedure complications may occur.
Once the embryos are collected into the sterile dish they are evaluated by our Embryologist. The evaluation process includes two main ratings called stage and grade. The embryos’ stage is determined by the point of development or maturity. The embryos’ grade is an evaluation of its quality or integrity. Once this is achieved, our Embryologist begins the embryo washing procedure to prepare them for transfer or freezing.
At this point, Dr. Thompson will laparoscopically evaluate the recipients to select the best candidate for embryo transfer. Once a recipient is selected, our Embryologist aids Dr. Thompson in the transfer of the embryos into the recipient. Two embryos per recipient are transferred unless specified by the client. Poorer quality embryos are paired with a higher quality embryo to increase transfer success.
The post-operative time period is very important to maintain a healthy stress-free environment. The recipients are to be turned into the same loafing barn with the same group and not handled for the next 10 days.
The process and preparation for semen collection should begin well in advance of the collection date. It is important to keep in mind the production of sperm cells begins 30 days prior to the day for your intended use. Therefore, it is VERY important that proper maintenance and management of the male is achieved 60 days prior to the planned usage. For more details please see pages focusing on animal preparation.
Semen collection should be performed to accurately evaluate semen quality prior to use in any of the many ram applications including natural breeding, laparoscopic artificial insemination (LAI), embryo transfer (ET) or for freeze or cryopreservation. Semen collection may be achieved by use of either an artificial vagina (AV) or via electroejaculation. Use of a ewe in estrus (teaser ewe) and collection with an artificial vagina (AV) is preferred. However, in the event a male is unwilling or unable to mount the female an electro-ejaculator (EJ) can be used with good success.
Once the semen sample is collected, it is evaluated to determine the concentration, percent motility, and morphologic status. First, the gross appearance of the ejaculate is evaluated based on the color, volume, and density. Next, the progressively motile sperm are examined. This is the percent of sperm that is motile in a straight line. Any sample under 70% is not suitable for AI unless extenuating circumstances exist. Finally, the sperm’s head, midpiece, and tails are observed to determine the sperm morphology. Abnormal morphology can have a major impact on proper sperm cell function, and as a result, negatively affect fertilization.
The males should be “cleaned out” (allowed to breed) 3-5 days prior to collection. This allows for the older sperm cells to be expelled from the male making way for more viable sperm cell.
Collecting males for frozen semen provides several benefits to an owner. Semen cryopreservation provides clients with insurance, peace of mind, increased marketability, and increased breeding management opportunities. The most important reason is that it serves as a form of insurance or protection for your breeding program in the event of the loss of the animal.
The collection of semen for freezing is maximized in late fall following the natural breeding season. As a result, our goal is to collect the majority of frozen semen between November and February. Collection dates are scheduled in advance in order to pool males together for more efficient handling. In some cases, frozen semen quality will improve over repeated collection days. In these cases, rams will be housed and collected on an every other day schedule. Reproduction Specialty Group provides services for housing and a facility for owners to haul to a central location for your convenience.
Once the semen is collected and evaluated, the sample is processed to begin cryopreservation. Collections not meeting a minimum standard will be discarded prior to the freezing process. Semen will be frozen in ¼ ml straws. A ¼ ml straw is preferred for the safety and security of proper identification. At Reproduction Specialty Group, ¼ ml straws are labeled using a laser printer that can include up to 90 characters. Following the freezing process, at least one straw is thawed and evaluated for quality. In the event the sample doesn’t meet our minimum post-thaw standard, the whole collection will be discarded and the male rescheduled for collection.
Once frozen, the straws are packaged and held in liquid nitrogen for long term storage. Quality frozen semen is required to have a minimum of 30 million progressively motile sperm cell per inseminant. The majority of our frozen semen will range from 60% to 75% progressively motile sperm cells. A post-thaw motility of 60% or less is discarded and the male should be recollected, unless under special circumstances recollection is not possible. The packaging of poor quality semen is a disservice to the owners and buyers of the semen. We strive to produce the absolute highest quality product for our clients.
Once frozen it is very important that the semen is stored in the liquid nitrogen. Nitrogen vapor is significantly warmer than liquid. The transition from cold to warm will cause damage to the cells through micro freeze/thaw. The same damage to the cell will occur from improper handling of semen while moving semen from tank to tank, splitting canes, or simple exposure to the air for more than one or two seconds. RSG provides storage options for your products to ensure the most effective storage.
Semen can be easily shipped through the use of our dry liquid nitrogen shippers. Semen is shipped via one of our traditional shipping companies or airlines and generally will arrive in one to two days.
For in-house semen collection date please contact Reproduction Specialty Group.
We have custom-built donor barn specially designed to enable us to host donors for optimum embryo production and careful monitoring. Call us to save your spot today!