Deep freezing of concentrated boar semen for intra-uterine insemination

Effects on sperm viability

Fernando Saravia, Margareta Wallgren, S. Nagy, Anders Johannisson, Heriberto Rodríguez-Martínez

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

The use of deep-frozen boar semen for artificial insemination (AI) is constrained by the need for high sperm numbers per dose, yielding few doses per ejaculate. With the advancement of new, intra-uterine insemination strategies, there is an opportunity for freezing small volumes containing high sperm numbers, provided the spermatozoa properly sustain cryopreservation. The present study aimed to concentrate (2 × 109 spz/mL) and freeze boar spermatozoa packed in a 0.5 mL volume plastic medium straw (MS) or a multiple FlatPack (MFP) (four 0.7 mL volume segments of a single FlatPack [SFP]) intended as AI doses for intra-uterine AI. A single freezing protocol was used, with a conventional FlatPack (SFP, 5 × 109 spz/5 mL volume) as control. Sperm viability post-thaw was monitored as sperm motility (measured by computer-assisted sperm analysis, CASA), as plasma membrane integrity (PMI, assessed either by SYBR-14/PI, combined with flow cytometry, or a rapid hypo-osmotic swelling test [sHOST]). Sperm motility did not differ statistically (NS) between test-packages and control, neither in terms of overall sperm motility (range of means: 37-46%) nor sperm velocity. The percentages of linearly motile spermatozoa were, however, significantly higher in controls (SFP) than in the test packages. Spermatozoa frozen in the SFP (control) and MFP depicted the highest PMI (54 and 49%, respectively) compared to MS (38%, P <0.05) when assessed with flow cytometry. In absolute numbers, more viable spermatozoa post-thaw were present in the MFP dose than in the MS (P <0.05). Inter-boar variation was present, albeit only significant for MS (sperm motility) and SFP (PMI). In conclusion, the results indicate that boar spermatozoa can be successfully frozen when concentrated in a small volume.

Original languageEnglish
Pages (from-to)1320-1333
Number of pages14
JournalTheriogenology
Volume63
Issue number5
DOIs
Publication statusPublished - Mar 15 2005

Fingerprint

Insemination
Semen
boars
insemination
Freezing
Spermatozoa
semen
freezing
spermatozoa
viability
Sperm Motility
Artificial Insemination
sperm motility
straw
artificial insemination
Sperm Count
dosage
Flow Cytometry
flow cytometry
Semen Preservation

Keywords

  • Boars
  • Computer-assisted sperm analysis (CASA)
  • Flow cytometry
  • High sperm count artificial insemination (AI) doses
  • sHOST
  • Spermatozoa

ASJC Scopus subject areas

  • Animal Science and Zoology
  • veterinary(all)

Cite this

Deep freezing of concentrated boar semen for intra-uterine insemination : Effects on sperm viability. / Saravia, Fernando; Wallgren, Margareta; Nagy, S.; Johannisson, Anders; Rodríguez-Martínez, Heriberto.

In: Theriogenology, Vol. 63, No. 5, 15.03.2005, p. 1320-1333.

Research output: Contribution to journalArticle

Saravia, Fernando ; Wallgren, Margareta ; Nagy, S. ; Johannisson, Anders ; Rodríguez-Martínez, Heriberto. / Deep freezing of concentrated boar semen for intra-uterine insemination : Effects on sperm viability. In: Theriogenology. 2005 ; Vol. 63, No. 5. pp. 1320-1333.
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abstract = "The use of deep-frozen boar semen for artificial insemination (AI) is constrained by the need for high sperm numbers per dose, yielding few doses per ejaculate. With the advancement of new, intra-uterine insemination strategies, there is an opportunity for freezing small volumes containing high sperm numbers, provided the spermatozoa properly sustain cryopreservation. The present study aimed to concentrate (2 × 109 spz/mL) and freeze boar spermatozoa packed in a 0.5 mL volume plastic medium straw (MS) or a multiple FlatPack (MFP) (four 0.7 mL volume segments of a single FlatPack [SFP]) intended as AI doses for intra-uterine AI. A single freezing protocol was used, with a conventional FlatPack (SFP, 5 × 109 spz/5 mL volume) as control. Sperm viability post-thaw was monitored as sperm motility (measured by computer-assisted sperm analysis, CASA), as plasma membrane integrity (PMI, assessed either by SYBR-14/PI, combined with flow cytometry, or a rapid hypo-osmotic swelling test [sHOST]). Sperm motility did not differ statistically (NS) between test-packages and control, neither in terms of overall sperm motility (range of means: 37-46{\%}) nor sperm velocity. The percentages of linearly motile spermatozoa were, however, significantly higher in controls (SFP) than in the test packages. Spermatozoa frozen in the SFP (control) and MFP depicted the highest PMI (54 and 49{\%}, respectively) compared to MS (38{\%}, P <0.05) when assessed with flow cytometry. In absolute numbers, more viable spermatozoa post-thaw were present in the MFP dose than in the MS (P <0.05). Inter-boar variation was present, albeit only significant for MS (sperm motility) and SFP (PMI). In conclusion, the results indicate that boar spermatozoa can be successfully frozen when concentrated in a small volume.",
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