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Evaluation of Fertilizing Potential of Frozen‐thawed dog Spermatozoa Diluted in ACP‐106 ® using an In Vitro Sperm–Oocyte Interaction Assay
Author(s) -
Cardoso RCS,
Silva AR,
Silva LDM,
Chirinéa VH,
Souza FF,
Lopes MD
Publication year - 2007
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/j.1439-0531.2006.00703.x
Subject(s) - semen , sperm , andrology , cryopreservation , membrane integrity , sperm motility , yolk , biology , oocyte , rose bengal , semen analysis , acrosome , chemistry , anatomy , membrane , biochemistry , embryo , food science , medicine , infertility , microbiology and biotechnology , pregnancy , genetics
Contents The aim of present study was to evaluate frozen canine semen with ACP‐106 ® (Powder Coconut Water) using an in vitro sperm–oocyte interaction assay (SOIA). Ten ejaculates from five stud dogs were diluted in ACP‐106 ® containing 20% egg yolk, submitted to cooling in a thermal box for 40 min and in a refrigerator for 30 min. After this period, a second dilution was performed using ACP‐106 ® containing 20% egg yolk and 12% glycerol. Samples were thawed at 38°C for 1 min. Post‐thaw motility was evaluated by light microscopy and by using a computer aided semen analysis (CASA). Plasma membrane integrity and sperm morphology/acrosomal status were evaluated by fluorescent probes (C‐FDA/PI) and Bengal Rose respectively. Moreover, frozen‐thawed semen was analysed by a SOIA. Subjective post‐thaw motility was 52.0 ± 14.8% and it was significant higher than the total motility estimated by CASA (23.0 ± 14.8%) because this system considered the egg yolk debris as immotile spermatozoa. Although normal sperm rate and acrosomal integrity evaluated by Bengal Rose stain was 89.6 ± 3.1% and 94.3 ± 3.1%, respectively, post‐thaw percentage of intact plasma membrane was only 35.1 ± 14.3%. Regarding SOIA, the percentage of interacted oocytes (bound, penetrated and bound and/or penetrated) was 75.3%. Using regression analysis, it was found significant relations between some CASA patterns and data for SOIA. In conclusion, the freezing‐thawing procedure using ACP‐106 ® was efficient for maintain the in vitro fertility potential of dog spermatozoa.