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Assessment of sperm attributes of frozen-thawed AI doses from Swedish and Estonian dairy bull sires

with special reference to pre-selection through swim-up, and the influence of age on potential fertility

Hallap, Triin (2005). Assessment of sperm attributes of frozen-thawed AI doses from Swedish and Estonian dairy bull sires. Diss. (sammanfattning/summary) Uppsala : Sveriges lantbruksuniv., Acta Universitatis agriculturae Sueciae, 1652-6880 ; 2005:113
ISBN 91-576-6912-0
[Doctoral thesis]

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Abstract

The fertility of bull semen used in artificial insemination (AI) is essential for the effective use of this technology in bovine breeding. The laboratory evaluation of semen from healthy bulls, before and after freezing for AI, is largely based on subjectively scoring sperm motility, and on measurement of sperm concentration. Since the relationship between these parameters is barely indicative of the fertility of the semen samples, other, often more complicated methods are used to evaluate a battery of sperm attributes of importance for their fertilizing capacity. This thesis evaluated the usefulness of various conventional and novel sperm quality tests in assessing frozen-thawed (FT) AI doses produced from semen of young, unproven and older, progeny-tested dairy bulls of the Estonian Holstein Friesian (EHF) and Swedish Red and White (SRB) breeds and their relationship with field fertility after AI. In addition, the influence of sperm preparation methods prior to quality measurements was investigated. Semen AI doses were evaluated immediately post-thaw (PT) and after cleansing through washing/resuspension (W) and swim-up (SU). Special attention was given to age-related changes in semen from 1- v. 4-, and 3- v. 5- and 7-year-old bulls. Spermatozoa were evaluated for their morphology (microscopy on wet and dried, stained smears), motility (assessed subjectively and by computer-assisted sperm analysis [CASA]) and membrane integrity (using SYBR-14/propidium iodide [PI] fluorophores and microscopy). Mitochondrial activity (with MitoTracker Deep Red), membrane fluidity (using Merocyanine 540 lipid dye/Yo-Pro-1/H332) and deoxyribonucleic acid (DNA) integrity (using acridine orange [AO] staining) were assayed using flow cytometry (FC). Use of SU provided spermatozoa with significantly better motility, morphology, membrane integrity, mitochondrial activity and chromatin stability compared with either PT or W treatment. Although the SU selection step increased the proportion of spermatozoa with a stable plasma membrane, it also initiated membrane destabilization. Age differences in sperm quality (motility, membrane integrity and mitochondrial function) were seen PT (at 4 years of age for SRB and at 3–7 years for EHF bulls) and were accentuated when using SU, but not W, as pre-selection procedure. No changes in chromatin stability were, however, registered for either breed. Only few FT sperm quality attributes (e.g. average path velocity (VAP), proportion of cells with tail abnormalities, and non-linear motility) showed significant correlation to fertility after AI when evaluated PT or after W. More attributes (e.g. CASA total sperm motility, concentration of motile and linearly motile spermatozoa, VAP, as well as the percentage of spermatozoa with unstable plasmalemma) had a significant relationship with non-return rate (NRR) when spermatozoa were examined after SU. In conclusion, SU was superior to W in harvesting intact spermatozoa with attributes essential for fertilization, distinguishing bulls and revealing age-dependent changes. Also, the use of CASA and FC of fluorophore-loaded spermatozoa increased the objectivity of the tests assayed. Combining SU and CASA, methods considered to be easily applicable at the semen-producing enterprises, indicated that the overall semen quality of proven bulls was predictable from measurements done at an early age. While sperm membrane stability and mitochondrial activity were seen as suitable markers for monitoring semen quality, measurements of chromatin intactness could not yield additional information.

Authors/Creators:Hallap, Triin
Title:Assessment of sperm attributes of frozen-thawed AI doses from Swedish and Estonian dairy bull sires
Subtitle:with special reference to pre-selection through swim-up, and the influence of age on potential fertility
Year of publishing :November 2005
Volume:2005:113
Number of Pages:33
Papers/manuscripts:
NumberReferences
ALLI. Hallap, T., Håård, M., Jaakma, Ü., Larsson, B., Rodriguez-Martinez, H. 2004. Does cleansing of frozen-thawed bull semen before assessment provide samples that relate better to potential fertility? Theriogenology 62, 702–713. II. Hallap, T., Håård, M., Jaakma, Ü., Larsson, B., Rodriguez-Martinez, H. 2004. Variations in quality of frozen-thawed semen from Swedish Red and White AI sires at one and four years of age. International Journal of Andrology 27, 166–171. III. Hallap, T., Jaakma, Ü., Rodriguez-Martinez, H. 2005. Changes in semen quality in Estonian Holstein AI bulls at 3, 5 and 7 years of age. Reproduction in Domestic Animals (accepted for publication). IV. Hallap, T., Nagy, Sz., Håård, M., Jaakma, Ü., Johannisson, A., Rodriguez-Martinez, H. 2005. Sperm chromatin stability in frozen-thawed semen is maintained over age in AI bulls. Theriogenology 63, 1752–1763. V. Hallap, T., Nagy, Sz., Jaakma, Ü., Johannisson, A., Rodriguez-Martinez, H. 2005. Mitochondrial activity of frozen-thawed spermatozoa assessed by MitoTracker Deep Red. Theriogenology 63, 2311–2322. VI. Hallap, T., Nagy, Sz., Jaakma, Ü., Johannisson, A., Rodriguez-Martinez, H. 2005. Usefulness of a triple fluorochrome combination Merocyanine 540/Yo-Pro-1/Hoechst 33342 in assessing membrane stability of viable frozen-thawed spermatozoa from Estonian Holstein AI bulls. Theriogenology (in press).
Place of Publication:Uppsala
ISBN for printed version:91-576-6912-0
ISSN:1652-6880
Language:English
Publication Type:Doctoral thesis
Full Text Status:Public
Agris subject categories.:L Animal production > L53 Animal physiology - Reproduction
Subjects:Not in use, please see Agris categories
Agrovoc terms:bulls, semen, spermatozoa, freezing, fertility, quality, laboratory experimentation, artificial insemination, sweden, estonia
Keywords:frozen-thawed (FT) bull semen, swim-up (SU), computer-assisted sperm analysis (CASA), flow cytometry (FC), in vivo fertility
URN:NBN:urn:nbn:se:slu:epsilon-800
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-800
ID Code:977
Department:(VH) > Dept. of Clinical Sciences
Deposited By: Triin Hallap
Deposited On:09 Nov 2005 00:00
Metadata Last Modified:02 Dec 2014 10:08

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