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Semen quality of stallions involved in breeding
VOKROUHLÍKOVÁ, Jana
The main objective of the study was to determine the seminal characteristics of warmblooded stallions, to assess a relationship between motility parameters of spermatozoa and sperm head dimensions (Experiment 1) and to compare the ability of three milk-based extenders to preserve the motility and viability of cooled shipped semen (Experiment 2) using Sperm Class Analyzer (SCA, MICROPTIC SL). In Experiment 1, 32 and 36 ejaculates were collected during breeding seasons 2016 and 2017 from 10 clinically healthy and fertile warmblooded stallions (age 3- 22 years). The volume of ejaculate (mL), sperm concentration (M/mL), motility of sperm with kinematic parameters, viability of sperm (%) and sperm morphometry with morphometric parameters were evaluated. In Experiment 2, 21 ejaculates from 11 stallions were collected before the beginning of the breeding season 2018 and diluted with Kenney, EquiPlus and INRA 96. Spermatozoa viability (VIT, %), total motility (MOT, %), progressive motility (PMOT, %) and selected kinematic parameters were evaluated for 63 semen samples. These parameters were determined 2 hours after semen processing and then in 24, 48 and 72-hour intervals of storage at 5 °C. The following average values were found in the obtained ejaculate samples for the observed periods of the breeding seasons 2016 and 2017 - volume of ejaculate 49.22 and 56.77 ml, sperm concentration 208.20 and 222.19 M/ml, MOT 82.44 and 81.38 %, PMOT 37.34 and 35.46 %, VIT 60.71 and 42.91 %, morphologically normal sperm 75.00 and 64.50 %, curvilinear velocity (VCL) 62.67 and 60.26 ?m/s, straight-line velocity (VSL) 32.10 and 27.55 ?m/s, average path velocity (VAP) 48.14 and 44.35 ?m/s, linearity coeficient (LIN) 50.64 and 44.89 %, straightness coeficient (STR) 66.56 and 62.01 %, wobble coeficient (WOB) 75.74 and 72.12 %, amplitude of lateral head displacement (ALH) 2.59 and 2.90 ?m, beat- cross frequency (BCF) 7.32 and 7.49 Hz, head length (L) 6.33 and 6,41 ?m, head maximum width (W) 3.01 and 3.11 ?m, head area (A) 15.65 and 16.37 ?m2, head perimeter (P) 16.26 and 16.55 ?m, elongation (Elong ) 0.36 and 0.35, ellipticity (Ellip) 2.11 and 2.07, roughness (Rugo) 0.74 and 0.75 and regularity (Regu) 0.96. There where significant differences among stallions in all evaluated parameters of sperm (P < 0.05). Furthermore, the influence of age and fertility of stallions was also demonstrated for the observed parameters. MOT, PMOT, VCL, VSL, VAP, LIN and WOB were positively correlated with area, width, length and perimeter of the head and negatively correlated with elongation and ellipticity of the sperm head. Sperms with larger head dimensions thus had larger MOT, PMOT, VCL, VSL, VAP, LIN, WOB, STR and smaller ALH and BCF. In contrast, sperms with greater elongation and ellipticity had lower motility and kinematic parameters. The effect of the extender (P < 0.05) and the storage time (P < 0.001) was highly significant for evaluated parameters. The ejaculates extended with INRA 96 reached significantly higher values (P < 0.01) for all evaluated parameters compared to Kenney extender. The INRA 96 extender significantly increased the values of PMOT, VCL and VAP compared to EquiPlus (P < 0.05). The results obtained from using INRA 96 showed that quality of cooled semen can be sufficiently maintained during at least 24 hours of storage.
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Hodnocení kvalitativních a kvantitativních ukazatelů ejakulátu u hřebců
Hájková, Ivana
Evaluation of semen quality is an important element in the evaluation of stallion fertility. Quality ejaculate produced in sufficient quantities is a prerequisite for successful insemination in horse breeding. To assess the quality of ejaculate is used by many laboratory evaluation methods. Selected methods are also engaged in this thesis. The basic parameters considerating in the ejaculate in this thesis are the evaluation of activity, sperm concentration and morphological sperm analysis. For using ejaculate in producing insemination doses need to be assessed volume of ejaculate too. Minimum volume of the ejaculate of stallions should be 20 ml. The activity of fresh semen should be at least 60 %. Normal sperm concentration value of stallions ejaculate is considered at least 100 × 106 in 1 ml of semen. The minimum amount of morphologically normal stallions sperm is required above the threshold value of 60 %. In this thesis we studied a group of stallions which were stabled in Zemský hřebčinec Tlumačov, s.p. Stallions were included in breeding season for 2015. Evaluation of quantitative and qualitative parameters were performed od 47 samples of fresh semen, which were collected from 12 stallions during the breeding season. Semen collecting was from April to July 2015. Immediately after each semen collecting was measured volume of ejaculate and sperm activity was recorded. To further evaluation were counting sperm concentration and morphology of sperm evaluation. For mophological analysis were preparations stained by two methods: B. T. Farelly staining and method Byghošť. The analysis was determined by the percentage of morphological sperm defects, which were divided into the following categories: acrosome defects, sperm head defects, sperm mid-piece defects, sperm tail defects, abnormal forms of sperm and immature sperm. The research also used a theoretical comparison of the two methods of stainings due to occurence of morphological defects and estimate the correlation relationships between qualitative and quantitative parameters of the ejaculated os stallions.
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