Breeding-induced endometritis in horses seems to be down-regulated by certain components in the seminal plasma. These components are most likely some of the proteins in the seminal plasma in the stallion ejaculate, the so-called spermadhesins. This study aimed to recognize these proteins and determine their quantities in different fractions of the ejaculate. Fractionated ejaculates from four stallions were used to determine the presence and amount of proteins by way of size-excluding chromatography. All former known proteins, except two, were recognized in this study.

Most Swedish Warmbloods are bred by artificial insemination (AI). Quality control of the semen consists of the subjective evaluation of motility. There is a need for a more accurate and objective method of rating the semen. In this study we compared fluorometry with flowcytometry as a method for evaluating sperm viability. Semen was collected from three stallions, stationed at Markebäcks Gård, Askersund, three times during week 15 and week 34, 2008.

Studiens målsättning var att utvärdera kvalitén hos Hingstsperma selekterad med hjälp av Single Layer Centrifugation (SLC), med huvudsyftet att studera den mitokondriella membranpotentialen (??m) och förekomsten av reaktiva syreföreningar (ROS) hos hingstspermier före och efter SLC. Ett insemineringsförsök med SLC selekterad och rutinspädd sperma genomfördes även på ett litet antal ston, men inga slutsatser kunde dras av resultaten. Humanstudier har visat att spermier med låg ??m mer frekvent har återfunnits hos män med fertilitetsproblem. Mätning av ??m har gjorts genom att tillsätta den lipofila JC-1, vilket i både studier på både humana- och ekvina spermier har visat sig vara en känslig metod för att särskilja spermier med låg respektive hög ??m.

There is considerable variation between stallions in semen quality and fertility due to the fact that selection of stallions for breeding is not based on fertility or semen quality but on performance. The increasing use of cooled semen for artificial insemination has resulted in decreasing pregnancy rates. Therefore the equine breeding industry needs new methods for evaluation and development of sperm quality and fertility. This study is a part of such a project and is performed by SLU in cooperation with Flyinge AB. Ten stallions at the Flyinge National Stud were used in the study and three ejaculates were collected from each stallion.

This study is part of a project to evaluate and develop metods to control the semen quality in stallions. The study was carried out in cooperation between SLU and Flyinge AB. The objective of the study was to analyse -if there are individual differences in sperm chromatin integrety between different stallions -if there is a correlation between subjectively evaluated sperm motility and sperm chromatin integrity -if sperm chromatin integrity is changed after selection of the semen with a centrifugation method through the silica-spheres, gradient or a single-layer, i.e to use the chromatin integrity analysis as a method to evaluate the selection method. The use of chilled transported semen has increased a lot during the last 10 years, especially among the Swedish warmblood breed. Because the foaling percentage has decreased with the increased use of chilled transported semen, there is a need to develop methods to control the sperm quality. Evaluation of the chromatin integrity could be one way to evaluate the quality of the semen. Chromatin is the DNA in the nucleus of the sperm.

This EEF-project is a part of a larger project at SLU, Division of Reproduction, Dept of Clinical Science, with the intention of developing and evaluating new methods for the assessment of stallion semen quality. Over the last 10 years there has been an increase in the use of artificial insemination with cooled semen in Swedish horse breeding. At the same time, foaling rates have decreased. This has lead to a need for new and better methods of assessing the quality of stallion semen. This EEF-project was based on an objective method to calculate motility, with QualispermTM, in fresh and cooled stallion semen, and after selection. Today sperm motility evaluation is mainly done subjectively by visual assessment with a light microscope, and the correlation between these motility results and fertility have varied.

The Annexin-V/propidium iodide (AV/PI) assay detects early membrane changes in spermatozoa and viability using flow cytometry. It is an objective method that evaluates a much larger number of single spermatozoa compared with the usual estimation of motility, the most commonly used method for routine sperm evaluation. In the present study four ejaculates from four stallions were used in a split-sample design to investigate whether and how the number of intact spermatozoa is affected over time and after incubation at different temperatures, either those recorded during shipment of stallion semen for AI (9 or 5°C) or room temperature (20°C). The Annexin-V results did not reflect the motility results that were monitored in parallel. While motility decreased over time, the number of spermatozoa with intact membranes (viable) remained unchanged for at least 21 hours following a short period of initial instability.

Vid transport av temperaturkänsliga produkter krävs en förpackning som klarar av att hålla temperaturen inom ett visst intervall så att innehållet inte skadas. Omgivningstemperaturen kan variera mycket under en transport och det är viktigt att förpackning kan kyla passivt utan att påverkas för mycket av omgivningen. Ett typiskt användningsområde för denna typ av förpackningar är inom sjukvård för transport av läkemedel, organ och andra biologiska produkter.Problemet som denna rapport behandlar är att ta fram en sådan förpackning för Hingstsperma som skulle kunna skickas med posten. Sperman behövde först kylas ned från rumstemperatur till 5°C under tio timmar innan den skulle hållas vid denna temperatur i ytterligare 24 timmar. För att lösa detta användes en kombination av isolering och fasomvandlingsmaterial vars egenskap att lagra energi gör att en konstant temperatur kan hållas under en längre tid.

Today, equine breeding is based on performance and conformation, resulting in a vast variation in fertility among different horses, with noticeable economic losses as a consequence. The horse breeding industry is therefore in need of methods that diagnose the spermatozoa concerning their quality and provide the best sperm for AI. The commonly used selection methods for sperm today are: centrifugation, where extender is added, the sample is centrifuged and then resuspended; swim-up self-migration; adherence separation; and density gradient centrifugation. However, none of these methods are in used routinely in practice before inseminating mares with fresh or cooled semen. The aim of this study was to evaluate a new selection method, Single Layer Centrifugation (SLC), and its effect on sperm viability. The viability was analyzed with regard to membrane integrity and membrane stability of ejaculated spermatozoa.

At SLU, Divison of Reproduction, Dept of Clinical Science, there is an ongoing project on the development and evaluation of new methods for the assessment of stallion semen quality. This EEF-study, which is a part of that larger project, has focused on the effects on motility and fertility of stallion semen of two different extenders and selection. Both an objective method and a subjective method for analysis have been used and compared in the study. The increase in the use of artificial insemination with cooled semen in Swedish horse breeding over the last decade is thought to play a part in declining foaling rates (Kareskoski et al, 2006, Kuisma et al, 2006). This calls for better evaluation methods of semen quality since cooling and transportation put higher demands on semen quality in order to maintain fertility (Aurich, 2005).