Semen cryopreservation is an essential biotechnology in canine reproduction and during the cryopreservation process commonly egg yolk are used. The discrepancy in the egg yolk composition and the potential risk of disease dissemination are obstacles for semen exportation and use. Therefore, studies aiming to substitute egg yolk are extremely important. In this context, soy lecithin contains a low-density lipoprotein fraction, is an interesting alternative. Thus, the objective of this study was to compare extenders based on soy lecithin (several concentrations and forms) with egg yolk during the cryopreservation process of dog sperm. For this purpose, we used twelve dogs. Semen was evaluated at different time points (after refrigeration, glycerolization, and thawing), by motility analysis (CASA) and functional tests (e.g., membrane integrity-eosin/nigrosin, acrosome integrity-fast green/Bengal rose, mitochondrial activity-3'3 diaminobenzidine, Chromatin susceptibility to acid-induced denaturation-SCSA, and susceptibility to oxidative stress-thiobarbituric acid reactive substances). The results indicated that egg yolk and lower concentrations of lecithin had similar effects on mitochondrial activity and motility. Thus, soy lecithin is a potentially viable alternative to egg yolk for the cryopreservation of dog semen.

Taurine bulls are highly susceptible to heat stress, leading to increased oxidative stress (OS) and impaired sperm viability. Polyunsaturated fatty acids (PUFAs) supplementation can be an alternative to improve semen quality, which also results in more sperm susceptibility to lipid peroxidation. Moreover, this deleterious effect can be exacerbated in animals affected by heat stress. Vitamin E is a key antioxidant that counteracts lipid peroxidation of sperm membrane caused by OS. Thus, combining PUFAs with vitamin E may improve sperm quality. In this context, this study aimed to evaluate the effect of interaction between PUFAs and vitamin E on sperm quality in Bos taurus bulls under testicular heat stress. Sixteen taurine bulls under testicular heat stress were randomly assigned in four groups: Control, Vitamin E, PUFA, and PUFA + Vitamin E. All groups lasted for 60 days. Samples were cryopreserved/thawed and analyzed for motility variables (CASA), membrane and acrosome integrity, mitochondrial activity, susceptibility to oxidative stress, DNA integrity, and sperm-binding capacity. Results showed that vitamin E had a beneficial effect on some sperm characteristics, whereas PUFA supplementation had an adverse effect when the two treatments were evaluated separately. Finally, the association between PUFAs and vitamin E did not improve sperm quality.

SummaryThe aim of this study was to compare different concentrations of soy lecithin (LEC0.01%, LEC0.05% and LEC0.1%) with egg yolk (Control) in cooling extenders during the storage of semen at 5ºC for 5 days. Twelve dogs (n = 12) were selected, and semen was cooled and assessed after 2, 24, 48, 72, 96 or 120 h. At each time point, sperm were analyzed for kinetic patterns (using computer-assisted sperm analysis), mitochondrial activity (3′3- diaminobenzidine assay), lipid peroxidation (TBARS assay), DNA fragmentation (SCSA®) and plasma and acrosome membrane integrity (eosin/nigrosin and fast green/rose Bengal stains, respectively). The Control group (1814.4 ± 197.2) presented the highest rates of lipid peroxidation at 120 h. Conversely, progressive motility (42.8 ± 4%), linearity (45.4 ± 1%), and VAP (88 ± 3%) were higher in the Control group. In addition, there was lower mitochondrial activity in the Control group at 72 h. Therefore, our data show that lecithin used at these concentrations was not able to maintain sperm viability at as high qualities as would egg yolk. Moreover, the decrease in high mitochondrial activity and the persistence of sperm motility may indicate a compensatory mechanism in canine spermatozoa (i.e., glycolytic pathway). Furthermore, these higher lipid peroxidation indexes could indicate the necessity for future therapy using extenders and antioxidants over a long cooling time for dog sperm.

Bos taurus bulls, when raised under tropical conditions, are highly susceptible to heat stress, which leads to impaired semen quality, leading to significant economical losses because, in these regions, the reproductive mounting season occurs mainly during the summer. Previous studies have indicated that oxidative stress (i.e. attack by reactive oxygen species) may be the main mechanism of sperm damage in such conditions. Therefore, treatment with antioxidants may be an important alternative to improve semen quality in heat-stressed B. taurus bulls. The objective of the present study was to evaluate whether the treatment with vitamin E, an important antioxidant, could improve sperm quality in insulated bulls. Towards this aim, eight adult Holstein bulls were submitted for semen collection, and the sperm was submitted for motility evaluation by computer-assisted sperm analysis (Ivos, Hamilton Thorne Inc., Beverly, MA, USA), examination of membrane and acrosomal integrity (eosin/nigrosin and fast green/bengal rose stain, respectively), mitochondrial activity (diaminobenzidine stain; full mitochondrial activity or no mitochondrial activity), and sperm susceptibility to oxidative stress (thiobarbituric acid-reactive substances). Bulls were then insulated (testicles covered in a thermal bag for 3 days) and randomly assigned to two treatment groups: no vitamin E (placebo) and vitamin E (subcutaneous injection of 3000 IU of α-tocopherol each of 10 days). Subsequent semen analysis was performed 1 and 60 days after the insulation. Statistical analysis was performed with SAS (SAS Institute Inc., Cary, NC, USA) repeated-measures ANOVA, and significance of P < 0.05 was adopted. No differences were found on any of the variables before insulation. One day after insulation, animals treated with vitamin E showed a lower percentage of static sperm and a higher percentage of motile sperm when compared with animals treated with the placebo (28 and 63% v. 56 and 34%, respectively; P < 0.05). Also at this time, sperm susceptibility to oxidative stress was lower in animals treated with vitamin E (vitamin E: 410 ng/106 sperm; no vitamin E: 1760 ng/106 sperm; P < 0.05). Sixty days after insulation, sperm susceptibility to oxidative stress was still lower in animals treated with vitamin E when compared with the placebo group (1176 and 192 ng/106 sperm, respectively; P < 0.05). However, no differences were found on the other variables. Results indicate that vitamin E, an antioxidant whose main function is protection of the plasma membrane, may be an alternative to avoid the acute deleterious effects of the heat stress in B. taurus bulls raised under tropical conditions. In addition, even with no heat stress involved, vitamin E treatment may provide constant protection, increasing the resistance of the sperm against the reactive oxygen species.

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