Concentration, motility and morphology are parameters commonly used to determine the fertilization potential of an ejaculate. These parameters give a general view on the quality of sperm but do not provide information about one of the most important components of the reproductive outcome: DNA. Either single or double DNA strand breaks can set the difference between fertile and infertile males. Sperm DNA fragmentation can be caused by intrinsic factors like abortive apoptosis, deficiencies in recombination, protamine imbalances or oxidative stress. Damage can also occur due to extrinsic factors such as storage temperatures, extenders, handling conditions, time after ejaculation, infections and reaction to medicines or post-testicular oxidative stress, among others. Two singular characteristics differentiate sperm from somatic cells: Protamination and absence of DNA repair. DNA repair in sperm is terminated as transcription and translation stops post-spermiogenesis, so these cells have no mechanism to repair the damage occurred during their transit through the epididymis and post-ejaculation. Oocytes and early embryos have been shown to repair sperm DNA damage, so the effect of sperm DNA fragmentation depends on the combined effects of sperm chromatin damage and the capacity of the oocyte to repair it. In this contribution we review some of these issues.

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This paper aimed at investigating the potential use of sperm DNA fragmentation (SDF) to improve the routine screening of infertility of Holstein bulls. Cryopreserved sperm samples from 201 Holstein bulls provided by an AI center were used in the analyses of SDF at 0 (SDF_0) and 6 (SDF_6) h of incubation at 37°C. A refinement of the sperm chromatin dispersion test implemented in the Sperm-Halomax kit was employed to measure SDF. Records on routinely collected semen traits (volume, concentration, mass and individual motility evaluated in the fresh ejaculate, and individual motility in post-thawed semen straws) were provided by the AI center. Artificial insemination bull fertility was obtained from official field recording as successful or failed insemination. The results show that the average SDF was low (around 3.5%) at 0 and 6 h of incubation. A moderate effect of inbreeding depression was found. Estimated heritability for SDF traits were moderately high (0.41 and 0.29 for SDF_0 and SDF_6, respectively) and estimated repeatability of SDF measures in the same animal were high (0.73 and 0.70 for SDF_0 and SDF_6, respectively). An overall estimated service bull value (ESBV) obtained through statistical modeling that allowed for adjustment of systematic environmental effects not specific to a bull and of the female contribution to fertility, and the estimated genetic values (EGV) were obtained from field-recorded AI information. The ESBV and EGV were also obtained for all semen traits. Moderately large and negative Pearson correlation coefficients were observed between SDF traits and male fertility ranging from (-0.43 to -0.50; P <0.001). Results of stepwise regression analyses showed that SDF_6 had the largest partial r(2) (0.15 to 0.26) among all semen characteristics. Overall, the selected semen traits explained 25% and 31% of the observed variability in bull fertility measured as EGV and ESBV, respectively. When looking at the predictive ability of bull fertility categories, the results of discriminant and logistic regression analyses showed that low-fertility bulls (those in the 10th or lower percentile in the fertility distribution) can be accurately identified by using measures of SDF alone or in combination with sperm motility. Values of SDF around 7% to 10% could be used as indicators of low AI success.

The gap in fertility between conventional and sex-sorted bovine sperm, known to be on the order of 10 percentage points, has never been bridged, even by increasing the number of sex-sorted sperm per inseminate. A concerted effort in the last few years has resulted in substantial changes in all stages of the sex-sorting process to develop an improved sex-sorted product called SexedULTRA™ (Sexing Technologies, Navasota, TX, USA). In vitro tests show that SexedULTRA™ maintained sperm integrity better than the previous XY method, and field trials were conducted to determine if this translated into improved bovine field fertility. The initial field trial was performed using ejaculates from 8 Holstein bulls located at Select Sires in Plain City (OH, USA). Each ejaculate was split in 2 aliquots that were then processed following the 2 methods (XY method or SexedULTRA™). A total of 6,930 Holstein heifers were inseminated across 41 commercial herds in the United States. Conception rate data were analysed using mixed model ANOVA (JMP 10.0.0; SAS Institute Inc., Cary, NC) with fixed effects of treatment, bull, bull by treatment interaction, and the random effect of herd. Sex-sorted bull sperm following the SexedULTRA™ method resulted in a greater (P < 0.001) conception rate compared with the XY method sperm (45.7 ± 1.7 v. 41.2 ± 1.6%). This is the first report of an improvement in conception rates using sex-sorted bovine semen in a decade since it became commercially available. A second field trial to investigate dose rate effects and compare against conventional semen was performed using 5 dairy bulls located at German Genetics International GmbH in Cloppenburg (Germany). Each ejaculate was split 4 ways, sex-sorted, and frozen in 0.25-mL straws as follows: XY 2.1 million/straw, SexedULTRA™ 2.1 million/straw, SexedULTRA™ 3.0 million/straw, and SexedULTRA™ 4.0 million/straw. A total of 7,855 heifers were inseminated with these sex-sorted straws, whereas 62,398 heifers were inseminated with conventional straws (15 million/straw) produced using contemporary ejaculates from the same bulls. The 56-day nonreturn rate was calculated by sire and treatment combination, and assigned a weight based on the total number of AI for each combination thereof. Table 1 contains the nonreturn rate weighted means and the relative fertility compared to conventional sperm. Data were analysed using a mixed model ANOVA (JMP 10.0.0; SAS Institute Inc.) with treatment and bull as fixed effects. SexedULTRA™ 4.0 sperm presented the greatest (P < 0.001) 56-day nonreturn rate followed by conventional 15.0 (66.73 v. 65.66%); XY 2.1 presented the lowest (P < 0.001) nonreturn rate. This is the first time a dose response effect with sex-sorted bovine sperm and parity in conception rates with conventional semen has been demonstrated. Table 1. Field trial results for number of inseminations, 56-day nonreturn rate (NRR) weighted means, and relative fertility1 compared with conventional sperm

This study examined DNA damage and postthaw motility of white-tailed deer sperm (n = 28) before and after sex selection and conventional sorting using MoFlo XDP SX flow cytometry. Semen samples from the same individuals were treated in 4 different ways: 1) chilled-extended sperm samples (without glycerol); 2) cryopreserved conventional samples, samples directly cryopreserved after the addition of extenders; 3) cryopreserved conventionally sorted samples, sorted samples to remove the dead sperm subpopulation; and 4) cryopreserved sex-sorted samples; sorted samples to remove the dead sperm subpopulation and separation of X- and Y-chromosome-bearing sperm. In all the cases (n = 6), conventional samples showed decreased postthaw motilities (43 ± 26%) when compared with X-sorted samples (59 ± 20%; P < 0.05) and Y-sorted samples (54 ± 20%; P > 0.05). The DNA fragmentation baseline was <5% for frozen-thawed conventional samples, but even less after sex sorting and conventional sorting: 2.4 and 1.7%, respectively. On the other hand, conventional samples showed greater (P < 0.05) DNA fragmentation than the sex-sorted sperm (n = 6) at 96 h (average of 4.8 ± 4.5% and 5.3 ± 4%, respectively). Conventionally sorted samples (n = 8) did not have greater (P > 0.05) DNA fragmentation when compared with the sex-sorted samples. Fragmentation of DNA on X-chromosome and Y-chromosome-bearing sorted sperm were not significantly different (n = 10, P > 0.05) after 96 h (2.6 ± 3.6% and 2.2 ± 0.5%, respectively). Future research should be implemented for examining the fertilizing potential of sex-sorted white-tailed deer sperm (e.g., AI fertility trials).