Thawing boar semen in the presence of seminal plasma: Effects on sperm quality and fertility.

Causes of poor fertility after insemination of frozen-thawed (FT) sperm include reduced sperm viability at thawing and a shorter longevity of surviving sperm in the female genital tract due to sub-lethal damage. The present studies examined the effect of incubating thawed boar sperm in seminal plasma (SP) on sperm membrane integrity (viability), and motility in vitro (experiment 1), and fertility in vivo (experiment 2). For experiment 1, FT sperm from five individual boars and a sperm pool from these boars were thawed and incubated for 4 h in media containing 0%, 10%, or 50% autologous seminal plasma (individual boars) or pooled seminal plasma (sperm pool). At approximately 10 min (0 h) and again at 1 h, 2 h, 3 h, and 4 h, sperm populations were examined for percentage sperm viability and percent sperm motility. Each variable progressively decreased during the incubation period. Incubation in 50% SP increased percentages of live sperm (P < 0.0001) and percent sperm motility (P < 0.01) at all time points compared to incubation in either 0% or 10% SP. For experiment 2, multiparous Large white x Landrace sows (n = 82) each received 900 IU eCG at weaning and 750 IU hCG 80 h later to control time of ovulation. Sows were assigned on the basis of parity to be inseminated with pooled semen with or without SP from the boars used in experiment 1. Sows received 3 x 10(9) live fresh-extended sperm (n = 30) or FT sperm thawed in 80 mL BTS extender (n = 26) or 3 x 10(9) live FT sperm thawed in 80 mL BTS containing 50% SP (FT-SP; n = 26). Sows were inseminated at 36 h, and 42 h after hCG injection. Compared to sows receiving fresh semen, the pregnancy rate of FT inseminated sows tended (P = 0.06) to be lower with the FT-SP group being intermediate. Farrowing rates were not different (83.3%, 69.2%, and 65.4% for fresh, FT, and FT-SP, respectively). Inseminations with FT sperm were associated with a reduction in litter size (P < 0.05), which was not evident in the FT-SP group. Taken together, these data confirm an adverse effect of inseminating FT sperm on sperm quality and sow fertility but suggest that thawing FT sperm in 50% SP may partially alleviate these adverse effects.

[1]  K. Weitze,et al.  Storage of boar semen. , 2000, Animal reproduction science.

[2]  K. Larsson,et al.  Fertility of Deep Frozen Boar Spermatozoa , 1976, Acta Veterinaria Scandinavica.

[3]  F. Peña,et al.  In vitro evaluation of the quality and fertilizing capacity of boar semen frozen in 0.25 ml straws. , 2006, Reproduction in domestic animals = Zuchthygiene.

[4]  T. Okazaki,et al.  Seminal plasma damages sperm during cryopreservation, but its presence during thawing improves semen quality and conception rates in boars with poor post-thaw semen quality. , 2009, Theriogenology.

[5]  T. Molitor,et al.  The effect of spermatozoa and seminal plasma on leukocyte migration into the uterus of gilts. , 1999, Journal of animal science.

[6]  D. Waberski,et al.  Effect of time of insemination relative to ovulation on fertility with liquid and frozen boar semen. , 1994, Theriogenology.

[7]  C. Ortega-Ferrusola,et al.  Detection of "apoptosis-like" changes during the cryopreservation process in equine sperm. , 2007, Journal of andrology.

[8]  S. Einarsson,et al.  A field trial on the fertility of deep-frozen boar spermatozoa. , 1973, Nordisk veterinaermedicin.

[9]  K. Roberts,et al.  Effects of seminal plasma on cooling-induced capacitative changes in boar sperm. , 2006, Journal of andrology.

[10]  M. Buhr,et al.  Effect of insemination-ovulation interval and addition of seminal plasma on sow fertility to insemination of cryopreserved sperm. , 2007, Reproduction in domestic animals = Zuchthygiene.

[11]  M. Vadnais,et al.  Effects of extender, incubation temperature, and added seminal plasma on capacitation of cryopreserved, thawed boar sperm as determined by chlortetracycline staining. , 2005, Animal reproduction science.

[12]  H. Rodríguez-Martínez,et al.  Comparative effects of autologous and homologous seminal plasma on the viability of largely extended boar spermatozoa. , 2004, Reproduction in domestic animals = Zuchthygiene.

[13]  J. Bailey,et al.  Impact of cryopreservation and reactive oxygen species on DNA integrity, lipid peroxidation, and functional parameters in ram sperm , 2007, Molecular reproduction and development.

[14]  H. Rodríguez-Martínez,et al.  Capacitation status and in vitro fertility of boar spermatozoa: effects of seminal plasma, cumulus-oocyte-complexes-conditioned medium and hyaluronan. , 2002, International journal of andrology.

[15]  G. Evans,et al.  Normal fertility in ewes after cervical insemination with frozen-thawed spermatozoa supplemented with seminal plasma. , 1999, Reproduction, fertility, and development.

[16]  M. Bernardi,et al.  Artificial insemination of gilts with 1.5 billion sperms stored in different periods associated with different pre-ovulatory intervals. , 2005, Reproduction in domestic animals = Zuchthygiene.

[17]  J. H. Rademaker,et al.  Use of boar spermatozoa for artificial insemination. III. Fecundity of boar spermatozoa stored in Beltsville liquid and Kiev extenders for three days at 18 C. , 1982, Journal of animal science.

[18]  L. Johnson,et al.  Viability and membrane integrity of spermatozoa after dilution and flow cytometric sorting in the presence or absence of seminal plasma. , 1996, Reproduction, fertility, and development.

[19]  D. Armstrong,et al.  Seminal plasma regulates endometrial cytokine expression, leukocyte recruitment and embryo development in the pig. , 2004, Reproduction.

[20]  S. Einarsson,et al.  Distribution of frozen-thawed spermatozoa in the reproductive tract of gilts at different time intervals after insemination. , 1973, Journal of reproduction and fertility.

[21]  J P Noordhuizen,et al.  Effect of superovulation induction on embryonic development on day 5 and subsequent development and survival after nonsurgical embryo transfer in pigs. , 2000, Theriogenology.

[22]  J. Calvete,et al.  Major proteins of boar seminal plasma as a tool for biotechnological preservation of spermatozoa. , 2008, Theriogenology.

[23]  H. Rodríguez-Martínez,et al.  Effect of freezing and thawing rates on the post-thaw viability of boar spermatozoa frozen in FlatPacks and Maxi-straws. , 2000, Animal reproduction science.

[24]  R. Tempelman,et al.  Effect of cooling and seminal plasma on the capacitation status of fresh boar sperm as determined using chlortetracycline assay. , 2005, Animal reproduction science.

[25]  R. Friendship,et al.  Effect of sperm cryopreservation and supplementing semen doses with seminal plasma on the establishment of a sperm reservoir in gilts. , 2007, Reproduction in domestic animals = Zuchthygiene.

[26]  J. Roca,et al.  Fertility of weaned sows after deep intrauterine insemination with a reduced number of frozen-thawed spermatozoa. , 2003, Theriogenology.

[27]  Leyland Fraser,et al.  Effect of different procedures of ejaculate collection, extenders and packages on DNA integrity of boar spermatozoa following freezing-thawing. , 2007, Animal reproduction science.

[28]  W. Holt,et al.  Sperm-oviduct interaction: induction of capacitation and preferential binding of uncapacitated spermatozoa to oviductal epithelial cells in porcine species. , 1999, Biology of reproduction.

[29]  H. Rodríguez-Martínez,et al.  Exposure to the seminal plasma of different portions of the boar ejaculate modulates the survival of spermatozoa cryopreserved in MiniFlatPacks. , 2009, Theriogenology.

[30]  D. Waberski,et al.  The initial fertilizing capacity of longerm-stored liquid boar semen following pre- and postovulatory insemination. , 1994, Theriogenology.

[31]  J. Calvete,et al.  Influence of Porcine Spermadhesins on the Susceptibility of Boar Spermatozoa to High Dilution1 , 2003, Biology of reproduction.

[32]  B. Kemp,et al.  In synchronized pigs, the duration of ovulation is not affected by insemination and is not a determinant for early embryonic diversity. , 1993, Theriogenology.

[33]  L. García-Marín,et al.  Changes in tyrosine phosphorylation associated with true capacitation and capacitation‐like state in boar spermatozoa , 2005, Molecular reproduction and development.

[34]  H. Rodríguez-Martínez,et al.  Sperm capacitation in the porcine oviduct. , 2004, Animal reproduction science.

[35]  R. Harrison,et al.  Use of fluorescent probes to assess membrane integrity in mammalian spermatozoa. , 1990, Journal of reproduction and fertility.