L-cysteine improves boar semen motility at 5 ºC but does not affect the oxidative status.

Hypothermic storage has been proposed as a method to reduce bacterial loads and promoting prudent use of antibiotics. Reducing temperature, however, can lead to cold shock damage and oxidative stress in boar semen. This study verified the effect of L-cysteine on the quality of semen stored at 5 °C for 120 h. Twenty-one normospermic ejaculates were diluted in Beltsville Thawing Solution into five treatments: Positive control (Pos_Cont, storage at 17 °C without L-cysteine) and groups with 0, 0.5, 1, and 2 mmol/L of L-cysteine supplementation stored at 5 °C. Variables were analyzed as repeated measures, considering treatment, storage time, and interaction as main factors. The effects of different L-cysteine concentrations were also evaluated using polynomial orthogonal contrasts. Sperm motility and pH were higher in the Pos_Cont compared to the groups stored at 5 °C (P < 0.05). In polynomial orthogonal contrast models, total motility was affected by the interaction between L-cysteine and storage time (P = 0.04), with a linear increase in motility when increasing the amount of L-cysteine at 72 and 120 h. Progressive motility increased quadratically as the L-cysteine reached 1 mmol/L (P < 0.01). In the thermoresistance test at 120 h, sperm motility increased quadratically up to an L-cysteine dose of 1 mmol/L (P < 0.05). Sulfhydryl content linearly increased with L-cysteine supplementation (P = 0.01), with no effect on intracellular ROS and sperm lipid peroxidation (P ≥ 0.06) in 5ºC-stored doses. In conclusion, L-cysteine supplementation has a positive effect on sperm motility up to 120 h of storage at 5 °C.

Sperm concentration of boar semen doses and sperm quality: Novel perspectives based on the extender type and sperm resilience.

This study evaluated the effect of sperm concentration of boar semen doses on their capacity to maintain its motility over the thermo-resistance test (TRT; sperm resilience) and verified if the extender type (short or long-term) could influence this effect. Thirty ejaculates from five crossbred mature PIC® boars were used, and a factorial design was followed to produce semen doses with 1.5 billion cells in 45 or 90 mL, using Beltsville Thawing Solution (BTS) or Androstar® Plus (APlus). Then, low-concentration doses (16.7 × 106 cells/mL in 90 mL) and higher-concentration doses (33.3 × 106 cells/mL in 45 mL) with BTS or APlus were produced and stored at 17 °C for 168 h. At 72 h, during the TRT, the low-concentration doses (16.7 × 106 cells/mL) lost three-fold less motility than doses with 33.3 × 106 cells/mL (P < 0.01), regardless of the extender type (11. 5% vs. 30.5% of initial motility, respectively). Similar results were found when the TRT was carried out at 168 h, with low-concentration doses losing two-fold less motility (11.4%) than highly concentrated doses (25.9%; P < 0.01). No sperm concentration effect was observed on membrane integrity or mitochondrial membrane potential (P ≥ 0.23). The osmolarity was not affected by the sperm concentration (P = 0.56), only by the extender and the storage time (P < 0.01). In conclusion, the sperm concentration effect on sperm quality was not influenced by extender type, and the data suggest that a low concentration of semen doses positively affects sperm resilience.