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Trypan Blue/Giemsa Staining to Assess Sperm Membrane Integrity in Salernitano Stallions and its Relationship to Pregnancy Rates
Author(s) -
Serafini R,
Longobardi V,
Spadetta M,
Neri D,
Ariota B,
Gasparrini B,
Di Palo R
Publication year - 2014
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/rda.12221
Subject(s) - sperm , andrology , acrosome , staining , sperm motility , trypan blue , biology , semen , membrane integrity , fertility , population , medicine , in vitro , biochemistry , genetics , environmental health , membrane
Contents Aim of this study was to test the reliability of T rypan blue/ G iemsa staining to evaluate sperm membrane integrity, acrosomal intactness and morphology in stallion to verify whether it could be applied in vitro as useful tool for sperm fertilizing ability. Fertility data on inseminated mares were collected to evaluate the relationship of sperm quality to pregnancy rates. Forty‐one ejaculates were collected from 3 stallions of S alernitano H orse B reed and evaluated for gross appearance, volume, visual motility and membrane integrity with T rypan blue/ G iemsa staining and thirty‐five mares were inseminated during the breeding season from A pril to J uly. Differences among stallions were found in volume, sperm concentration (p < 0.05) and visual motility (p < 0.01). A decrease in sperm motility, concentration (p < 0.05) and total sperm number was found in June– J uly (p < 0.01). Live sperm with intact acrosome ( LSIA ) and proximal droplets ( PD ) were lower (p < 0.01) in June–July, while acrosome reacted sperm ( ARS ) percentage increased (p < 0.05). No fertility differences were found among stallions with an average fertility per cycle of 44.6% and a pregnancy rate of 68.6%. Higher percentages of LSIA were found in the ejaculates used to inseminate mares that became pregnant vs those used in mares not pregnant (p < 0.05). The significance of LSIA as test variable to verify the reliability of T rypan blue/ G iemsa staining was confirmed by Receiver operating characteristic ROC analysis and the sensitivity of the test was 85% at a cut‐off value of 48% LSIA . Trypan blue‐ G iemsa showed to be an accurate method that can be applied on field to evaluate sperm membrane integrity and to identify poor‐quality ejaculates.