Motility is one of the most important characteristics associated with the fertilising ability of spermatozoa indicating their viability and structural integrity. Therefore, the examination of motility constitutes an integral part of semen analysis. Computer-assisted semen analysis (CASA) allows an accurate and objective assessment of different sperm motion characteristics with high repeatability. The aim of this study was to evaluate the different kinematic (velocity) parameters of frozen/thawed bull semen and determine if any of them could be correlated with their fertilising capability after insemination based on the achieved pregnancy rate. Ejaculates from 10 bulls were collected and frozen. The kinematic/velocity parameters of spermatozoa were measured by CASA and compared to the pregnancy results of almost 9,000 females artificially inseminated (AI) with frozen semen of any of the 10 tested bulls. The data of the experiments are summarised mainly with a focus on the effects of individual velocities (curvilinear velocity: VCL, straight-line velocity: VSL, average path velocity: VAP) on fertility rather than on the influence of progressive motility as a whole. We conclude that VAP is the most useful semen motility characteristic which has clinical relevance in the prediction of fertility.Key words: Semen evaluation, CASA, fertility prediction, velocity characteristics, semen motility Male fertility is an important factor in bovine reproduction, considering that the semen of any particular bull will be used for the insemination of several females. Fertility evaluation of males is primarily based on quality assessment of semen using conventional parameters such as morphology, concentration and motility of spermatozoa. The correlation between semen quality and fertility has been well documented (Bratton et al., 1956;Davis et al

We developed a method to evaluate bovine sperm membranes in normal (1G) and simulated microgravity (Sim-µG). Bovine spermatozoa are used as a model system because they have cellular membranes analogous to those of other cell types, and yet are much simpler because they have no cytoplasm and do not participate in DNA transcription or mRNA translation. They can be cultured as single cells and are easily evaluated for membrane characteristics using flow cytometry. These features make the mammalian spermatozoon a useful model for exploring the principles of membrane structure/function in the presence of a variety of environmental challenges such as simulated microgravity. Cryopreserved, washed beef bull sperm (4-8 × 106 mL −1 ) were incubated under non-capacitating conditions (modified glucose-free Tyrode's medium containing low bicarbonate, HEPES buffer, pyruvate and 3 mg mL −1 BSA V; 23 • C in air), and these spermatozoa remained alive for 24-48 h at 1G. To simulate µG, spermatozoa were incubated under the same conditions, in a HARV 10 rotating wall vessel (RWV, Synthecon, Inc, Houston,TX, USA) at 9 rpms. Spermatozoa were incubated in 1G and Sim-µG environments for 2.5-4.5 h and subsequently exposed to 0, 60 or 80 µg mL For further statistical analysis, and incorporation of non-parametric statistical tools (including pattern recognition using Support Vector Machines), the data were processed using a collection of Perl scripts and C programs. Results: Live/dead status: When Sim-µG + 60 µg mL −1 LC sperm were compared to 1G + 60 µg mL −1 LC, and 80 µg mL −1 LC sperm, their profiles were more similar to the 1G 80 µg mL −1 LC profiles. AR status: the Sim-µG + 60 µg mL −1 LC profiles were similar to the 1G + 60 µg mL −1 LC profiles. Mitochondrial Status: the Sim-µG + 60 µg mL −1 LC profiles were more similar to 1G + 80 µg mL −1 LC profiles. Summary: although Sim-µG sperm lost their motility within 3 h, they were alive. Cell profiles indicate that Sim-µG sperm nuclear membranes are less stable and their mitochondria are less functional than the 1G controls, but their acrosomes are intact indicating that fertilizing potential may remain. Additional experiments are needed to determine the time course for Sim-µG, induced changes, and whether Sim-µG sperm can penetrate eggs.