Effect of topical vaginal products on the detection of prostate-specific antigen, a biomarker of semen exposure, using ABAcards
Margaret Christine SneadAthena P. KourtisCarolyn M. BlackChristine K. MauckTeresa M. BrownAna Penman‐AguilarJohan H. MelendezMaria GalloDenise J. JamiesonMaurizio Macaluso
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Despite several efforts to standardize methods of semen analysis, sperm count is known to be subject to large interlaboratory differences. This is especially a problem in multicentre clinical trial settings and protocols for the preparation of semen for centralized assessment of sperm concentration are suggested here. The stability of semen has been tested after fixation with formalin at different dilutions and at different temperatures for different sperm concentrations. Sperm concentrations in formalin-fixed semen (at dilutions 1 + 0.5 to 1 + 19) were stable (<20% difference from original) at all concentrations (0.1 x 10(6)/mL to 100 x 10(6)/mL) for at least 5 days at room temperature and 4 degrees C. Prolonged stability up to 5 weeks at 4 degrees C was demonstrated for the lower dilutions (1 + 0.5 and 1 + 1). Freezing of fixed semen samples was unacceptable. These results illustrate that centralized assessment of sperm concentrations is feasible.
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Indonesia has two National Artificial Insemination centers and 17 Regional Artificial Insemination Centers. The frozen semen production techniques differed between the centers, including the type of diluent and semen dilution technique. The aim of the research was to compare the quality of frozen Limousin bull semen diluted using different techniques.Semen was collected from three sexually mature Limousin bulls using an artificial vagina. Immediately after collection, the semen was evaluated macroscopically and microscopically. Semen that had >70% motile sperm and <20% sperm abnormality was divided into three tubes and diluted with skim milk-egg yolk (SMEY) using three different dilution techniques: One-step dilution (100% SMEY with 8% glycerol) at room temperature ([RT] 20°C until 25°C) two-step dilution (50% SMEY without glycerol at RT, stored at 5°C; and 50% SMEY with 16% glycerol after 1 h stored at 5°C); and three-step dilution (50% SMEY without glycerol at RT, stored at 5°C; and 50% SMEY with 16% glycerol added twice at 1 h and 1.5 h after being stored at 5°C). The diluted semen was loaded into 0.25 mL mini straws, equilibrated, and frozen using a freezing machine. Sperm motility, viability, membranes, DNA integrity, and concentrations of malondialdehyde (MDA) and aspartate aminotransferase (AST) enzymes were evaluated after thawing.The results showed that there were no significant differences in sperm motility and DNA integrity between dilutions (p>0.05). However, sperm viability and membrane intactness of one-step dilutions were higher than those of three-step dilutions. The concentrations of MDA and AST enzymes of sperm in one-step dilutions were lower than those of three-step dilutions (p<0.05).It was concluded that the one-step-dilution technique was better than three-step dilution for cryopreservation of Limousin bull semen.
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This study examined factors affecting the analysis of motility of chicken semen. The viability of spermatozoa was estimated using varying dilution ratios and supplementation with BSA or fatty acid free (FAF)-BSA as protein sources in semen diluent. Fresh semen was examined after preparing dilutions in beltsvile poultry semen extender (BPSE) of 1/8, 1/16 and 1/32 at 25℃. The motility of incubated semen at each dilution was observed at 3 min (89.9%, 69.9% and 53.2%), 30 min (86.7%, 71.4% and 51.7%), 1 h (89.5%, 74.0% and 53.5%) and 3 h (78.5%, 66.5% and 45.7%), respectively. The addition of BSA or FAF-BSA to BPSE diluent significantly increased the viability of semen in 1/32 dilution with results of 53.2% (control), 84.8% (BSA) and 92.9% (FAF-BSA) (p<0.05). This phenomenon was also observed in the dilution of frozen semen, where FAF-BSA treatment increased the viability of thawed semen from 17.6% to 34.0% in a 1/8 dilution (p<0.05). When the protein sources were used in the dilution, the survival rates of diluted chicken semen were also increased with time lapse. These results show that FAF-BSA may act to protect chicken semen and is suitable as a basic component of chicken semen diluent for the method of analyzing rooster semen after freezing.
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