Conventional Semen Research Articles

Postnatal phenotype of dairy cows is altered by in vitro embryo production using reverse X-sorted semen

Abnormal fetuses, neonates, and adult offspring derived by assisted reproductive technologies have been reported in humans and mice and have been associated with increased likelihood of certain adult diseases. To test the hypothesis that bovine females derived by assisted reproductive technologies have altered postnatal growth and adult function, a retrospective cohort study evaluated survival, growth, and production traits of offspring derived by in vitro embryo production (IVP) with conventional (IVP-conv) or reverse X-sorted semen (IVP-sexed), multiple ovulation and embryo transfer, and artificial insemination (AI) in a large dairy herd. Live calves produced by IVP were born slightly heavier compared with AI calves. In addition, IVP-sexed calves had a higher cumulative mortality from 90 to 180 d of age compared with AI offspring. Mortality of IVP-conv and multiple ovulation and embryo transfer offspring was intermediate and not different from AI or IVP-sexed offspring. The altered phenotype of offspring from IVP-sexed extended to adult milk production. Cows derived by IVP-sexed produced less milk, fat, and protein in their first lactation compared with dairy cows derived by AI. Additionally, females born to nulliparous dams had a distinct postnatal phenotype compared with offspring from parous dams even when data were restricted to offspring of surrogate females. In conclusion, procedures associated with in vitro production of embryos involving use of reverse-sorted spermatozoa for fertilization result in an alteration of embryonic programming that persists postnatally and causes an effect on milk production in adulthood. Thus, some benefits of reverse-sorted semen for genetic improvement may be offset by adverse programming events.

MP35-06 STATIN USE IS NOT DETRIMENTAL TO SEMEN PARAMETERS IN SUBFERTILE MEN

You have accessJournal of UrologyInfertility: Epidemiology & Evaluation II1 Apr 2017MP35-06 STATIN USE IS NOT DETRIMENTAL TO SEMEN PARAMETERS IN SUBFERTILE MEN James M. Hotaling, Sorena Keihani, James R. Craig, Chong Zhang, Angela P. Presson, Jeremy B. Myers, William O. Brant, Kenneth I. Aston, Benjamin R. Emery, and Douglas T. Carrell James M. HotalingJames M. Hotaling More articles by this author , Sorena KeihaniSorena Keihani More articles by this author , James R. CraigJames R. Craig More articles by this author , Chong ZhangChong Zhang More articles by this author , Angela P. PressonAngela P. Presson More articles by this author , Jeremy B. MyersJeremy B. Myers More articles by this author , William O. BrantWilliam O. Brant More articles by this author , Kenneth I. AstonKenneth I. Aston More articles by this author , Benjamin R. EmeryBenjamin R. Emery More articles by this author , and Douglas T. CarrellDouglas T. Carrell More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.1096AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Data from animal studies and small trials show conflicting results regarding the effects of statins on semen quality. Evidence is even more limited in subfertile patients. We aimed to assess the effects of statin use on semen parameters. To investigate this, we retrospectively compared semen parameters in statin users vs. non-users in subfertile men. METHODS From 2002-2013, we reviewed data from 12257 subfertile men visited at our fertility clinic. Patients who reported using any statin drugs for >3 months before semen sample collection were identified. Data on patient age, medication use, and conventional semen parameters were extracted. 7698 subfertile men taking no medications served as controls. Patients who were using any known spermatotoxic medications were excluded from the study. Variables with non-normal distributions (concentration, total count, and total motile sperm count) were log-transformed for analyses and the corresponding coefficients were presented as ratios. Linear mixed effect regression models were used to test the effects of statin use on semen parameters adjusting for age. The model coefficients, 95% confidence intervals (CIs) and p-values were reported, and statistical significance was assessed at the 0.05 level. RESULTS A total of 109 patients were identified taking statins. Mean age was 38.3 (standard deviation: 7.3). Estimates from the age-adjusted regression model are presented in Table-1. Ejaculate volume was 0.3 ml lower (95% CI: -0.58, -0.02 ml, p-value=0.04) and sperm viability was 4.52% higher (CI: 1.65%, 7.39 %, p-value=0.002) in statin users compared to controls. CONCLUSIONS Statin use was not adversely associated with semen parameters other than semen volume in subfertile patients. However, this should be interpreted cautiously given the limited sample size and retrospective nature of this study. Additional studies are needed to confirm these findings. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e459 Advertisement Copyright & Permissions© 2017MetricsAuthor Information James M. Hotaling More articles by this author Sorena Keihani More articles by this author James R. Craig More articles by this author Chong Zhang More articles by this author Angela P. Presson More articles by this author Jeremy B. Myers More articles by this author William O. Brant More articles by this author Kenneth I. Aston More articles by this author Benjamin R. Emery More articles by this author Douglas T. Carrell More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Effective use of SexedULTRA™ sex-sorted semen for timed artificial insemination of beef heifers

An experiment was designed to evaluate the relative fertility of SexedULTRATM sex-sorted semen compared to conventional, non-sex-sorted semen when used among beef heifers in conjunction with split-time AI following the 14-d CIDR-PG protocol. Units of conventional semen were generated with 25.0 × 106 live cells per 0.5 ml straw prior to freezing, and units of sex-sorted semen were generated using the SexedULTRATM Genesis III sorting technology with 4.0 × 106 live cells per 0.25 ml straw prior to freezing. Sex-sorted units were sorted to contain X chromosome-bearing sperm cells at an accuracy level of >90%. Estrus was synchronized in 851 heifers at four locations using the 14-d CIDR-PG protocol: controlled internal drug release (CIDR) insert (1.38 g progesterone) on Day 0, CIDR removal on Day 14, and administration of prostaglandin F2α (PG; 25 mg im) on Day 30. Estrus detection aids were applied at PG on Day 30 to evaluate estrous response rate, and split-time AI was performed based on estrous response. At 66 h after PG (Day 33), heifers having expressed estrus received timed AI. Heifers failing to express estrus by 66 h received timed AI 24 h later (90 h after PGF2α on Day 34). Heifers failing to express estrus by 90 h were administered gonadotropin-releasing hormone (GnRH; 100 μg im) concurrent with AI. Heifers were preassigned to treatment (insemination with either conventional or SexedULTRATM sex-sorted semen), and treatments were balanced within each location based on source, reproductive tract score, and weight. Heifers were exposed for natural service beginning 14 d after AI for the remainder of a 60 d breeding season. Pregnancy rates to AI across locations tended to be higher (P = 0.09) for heifers inseminated with conventional semen (60%; 257/429) compared to sex-sorted semen (52%; 218/422). Higher pregnancy rates to AI (P < 0.0001) were obtained among heifers that expressed estrus prior to AI than among heifers that failed to express estrus prior to AI at 90 h. Total pregnancy rates at the end of the 60 d breeding season did not differ between heifers that received sex-sorted semen at AI (89%; 376/422) and heifers that received conventional semen at AI (89%; 382/429). In summary, the pregnancy rates observed suggest that SexedULTRATM sex-sorted semen can be used effectively for timed AI of beef heifers when split-time AI is performed following the 14-d CIDR-PG protocol.

Short communication: Optimization of a timed artificial insemination program for reproductive management of heifers in Canadian dairy herds

Pregnancy per AI (P/AI) following the use of 1 of 2 timed AI (TAI) protocols and 2 different intervals between TAI and resynchronization were compared in heifers that were inseminated with either conventional or sex-sorted semen. Holstein heifers (n = 317; 527 inseminations) were submitted to a 5-d Cosynch protocol with (+) or without (-) GnRH at the time of controlled internal drug release (CIDR) insertion on d 0, CIDR removal and a single PGF2α treatment on d 5, and TAI plus GnRH on d 8 (72 h later). Visual estrus detection (ED) was conducted on d 6 in the afternoon and d 7 in the morning and heifers observed in estrus were artificially inseminated on d 7 in the afternoon. Heifers were alternately assigned conventional or sex-sorted semen. Pregnancy was diagnosed by ultrasound 27 and 42 d after AI, and heifers diagnosed as nonpregnant were resynchronized, up to 3 times, starting on d 27 or 34 to provide an interbreeding interval of 35 or 42 d. Overall, TAI protocol had no effect on P/AI at 27 or 42 d after artificial insemination or on pregnancy loss, but P/AI following the first service tended to be higher in the -GnRH TAI group (66.3 vs. 56.8%). Pregnancy per AI at 27 d (61.9 vs. 55.5%) tended to differ between conventional and sex-sorted semen. Heifers artificially inseminated based on ED tended to have a greater P/AI (67.6 vs. 58.2%) and had decreased pregnancy loss (0.0 vs. 4.1%) than those submitted to TAI. A greater number of heifers in the -GnRH TAI protocol were artificially inseminated on ED than the +GnRH TAI protocol (21.5 vs. 13.7%). No difference in P/AI was observed between the 35- and 42-d interbreeding intervals; however, more heifers in the 42-d group were artificially inseminated based on ED than in the 35-d group (22.7 vs. 7.8%). A 5-d Cosynch+CIDR TAI protocol without the initial GnRH and with a single PGF2α at CIDR removal is an acceptable alternative to achieve high P/AI when either conventional or sex-sorted semen is used in Holstein heifers. Breeding heifers based on detected estrus increases labor, but has the potential to increase fertility.

Use of the fluorescent dye tetramethylrhodamine methyl ester perchlorate for mitochondrial membrane potential assessment in human spermatozoa

Mitochondrial membrane potential (ΔΨm) is an indicator of sperm quality and its evaluation complements the standard semen analysis. The fluorescent dye JC-1 has been widely used to assess sperm ΔΨm; however, some problems have been detected under certain experimental conditions. Another fluorescent compound, tetramethylrhodamine methyl ester perchlorate (TMRM), has been used in somatic cells and bovine spermatozoa but not in human spermatozoa. TMRM accumulates in hyperpolarised mitochondria and the fluorescence intensity of this compound correlates with ΔΨm. Thus, the aim of this study was to evaluate and validate the usefulness of the fluorescent dye TMRM for measuring sperm ΔΨm. The results showed that TMRM accurately detects sperm populations displaying either high or low ΔΨm. Moreover, TMRM was able to measure sperm ΔΨm under the experimental conditions in which JC-1 had previously presented difficulties. Differences in ΔΨm according to sperm and semen quality were properly detected and a positive correlation between ΔΨm and conventional semen parameters was observed. Finally, a positive correlation was found between the ΔΨm measurement by TMRM and by the widely used JC-1. In conclusion, TMRM is a simple, time-effective method, easy to set in laboratories equipped with flow cytometry technology, and can accurately detect changes in ΔΨm with efficiency comparable to JC-1 without its limitations.

Understanding farmers’ preferences for artificial insemination services provided through dairy hubs

Africa has a shortage of animal products but increasing demand because of population growth, urbanisation and changing consumer patterns. Attempts to boost livestock production through the use of breeding technologies such as artificial insemination (AI) have been failing in many countries because costs have escalated and success rates have been relatively low. One example is Kenya, a country with a relatively large number of cows and a dairy industry model relevant to neighbouring countries. There, an innovative dairy marketing approach (farmer-owned collective marketing systems called dairy hubs) has been implemented to enhance access to dairy markets and dairy-related services, including breeding services such as AI. So far, the rate of participation in these dairy hubs has been slow and mixed. In order to understand this phenomenon better and to inform dairy-related development activities by the Kenyan government, we investigated which characteristics of AI services, offered through the dairy hubs, farmers prefer. To do so, we applied a choice experiment (CE), a non-market valuation technique, which allowed us to identify farmers’ preferences for desired characteristics should more dairy hubs be installed in the future. This is the first study to use a CE to evaluate breeding services in Kenya and the results can complement findings of studies of breeding objectives and selection criteria. The results of the CE reveal that dairy farmers prefer to have AI services offered rather than having no service. Farmers prefer AI services to be available at dairy hubs rather than provided by private agents not affiliated to the hubs, to have follow-up services for pregnancy detections, and to use sexed semen rather than conventional semen. Farmers would further like some flexibility in payment systems which include input credit, and are willing to share the costs of any AI repeats that may need to occur. These results provide evidence of a positive attitude to AI services provided through the hubs, which could mean that AI uptake would improve if service characteristics are improved to match farmer preferences. The dairy hubs concept is currently in the implementation phase with most hubs at startup phase, hence understanding which AI service characteristics farmers prefer can inform the design of high-quality and cost-effective AI services in the future.

Sex-sorted sperm in farm animals: scale of use, the effectiveness of insemination, risk factors

The aim of the study was to present the scale of use, risk factors and possibilities, which sorter semen gives in biotechnics used in reproduction of cattle. Modern sorters allow for the evaluation of 6 million X and Y spermatozoa per hour. Sex-sorted semen, which is commercially used, contains 2.1 x 106 of spermatozoa. It is used mostly in AI of milk heifers, mainly in large cattle herds. Sorted semen containing Y spermatozoa is sold less often in the world than the one with X spermatozoa. The percentage of the desired sex of the young is higher than 90. The pregnancy rate after application of sorted semen is about 20–25% lower than after insemination of non-sorted semen and depends on a number of factors. The main factors are: breed of female, service number, the herd of origin, the depth of semen deposition, the bull producing semen, ambient temperature and technical parameters during sperm sorting. A number of methods have been developed to improve conception rate, including timed artificial insemination (TAI) and synchronization of heat and ovulation. Results of donor inseminations with the use of sorter semen are presented, with the lower percentage of embryos suitable for the transfer and embryos of the highest quality highlighted. Previous studies do not indicate a reduction of the conception rate after the transfer of embryos obtained in vitro and in vivo after fertilization using sorted semen. It remains difficult to justify a significant increase in the frequency of stillbirths of bulls after using sorted sperm. Similarly, 16% of stillbirths of bulls were observed after embryo transfer, when donors were inseminated with sorter semen. The percentage of stillbirths of bulls after embryo transfer with the use of conventional semen is 9%. The sorted semen is not often used for inseminations in pigs, sheep and goats.

Influence of the abstinence period on human sperm quality: analysis of 2,458 semen samples.

ObjectiveThe aim of this study was to evaluate the influence of different periods of abstinence on conventional semen parameters as well as functional parameters in human semen, including mitochondrial function, chromatin packing and sperm DNA fragmentation.MethodsWe recruited a cohort of 2,458 men undergoing infertility investigation. Semen analyses were performed according to WHO guidelines/morphology-motile sperm organelle morphology examination/MSOME. For DNA integrity analysis, the percentages of DNA fragmentation (TUNEL), abnormal chromatin packaging/underprotamination (chromomycin A3/CMA3), abnormal mitochondrial membrane potential (MMP/MitoTracker Green), and apoptosis (annexin-V) were recorded. Associations between the sexual abstinence period and sperm parameters were assessed using Spearman correlation. For group comparisons, the subjects were categorized according to the sexual abstinence period (SAP) into three groups: SAP <2 days, SAP 2-5 days, and SAP >5 days.ResultsThe duration of abstinence had a statistically significant positive influence on sperm concentration and volume, the number of leukocytes and a statistically significant negative influence on sperm motility and vitality. The percentages of DNA fragmentation and MMP (mitochondrial damage) worsened with the increased duration of abstinence. The percentage of sperm protamination was statistically significantly increased with abstinence.ConclusionIncrease in the sexual abstinence period influences sperm quality. This study reinforces the importance of the duration of ejaculatory abstinence on semen parameter variation. It highlights the deleterious effect of increased abstinence on DNA damage, which is most likely associated with ROS (mitochondrial damage/number of leukocytes). The increase in chromatin packaging can represent a protective feature for DNA.

Semen parameters in men with varicocele: DNA fragmentation, chromatin packaging, mitochondrial membrane potential, and apoptosis.

ObjectiveTo evaluate varicocele effects on conventional semen parameters: sperm DNA fragmentation, chromatin packaging, mitochondrial membrane potential (MMP) and sperm apoptosis.MethodsA cross-sectional study was carried out with semen samples from 2,399 men of couples who attended an infertility clinic. A total of 16.3% (391/2399) of the men were diagnosed with varicocele by a urologist.ResultsA regression analysis revealed that the percentages of sperm with DNA fragmentation, abnormal chromatin packaging, and abnormal MMP were significantly increased in individuals with varicocele, when compared to men without varicocele. Apoptosis was not influenced by varicocele. Conventional semen parameters were significantly worse in individuals with the disease. On the other hand, in men with varicocele, Spearman's correlation demonstrated that early apoptosis and abnormal MMP showed a positive and significant correlation with sperm DNA fragmentation.ConclusionMen with varicocele had worse semen parameters, including increased levels of sperm DNA fragmentation, inactive mitochondria, and abnormal chromatin packaging. These changes are possible causes of infertility in individuals with varicocele.

Mitochondrial Biomarkers Reflect Semen Quality: Results from the MARCHS Study in Chongqing, China.

Unexplained infertility requires that more sensitive and mechanism-based biomarkers should be developed and used independently of or in addition to conventional semen parameters for an infertility diagnosis. In the present study, semen samples were collected from young men participating in the Male Reproductive Health in Chongqing College students (MARCHS) cohort study in the follow-up stage in 2014. Conventional semen parameters were measured in all 656 participants, whereas sperm mitochondrial membrane potential (MMP), mitochondrial DNA copy number (mtDNAcn), mtDNA integrity and apoptotic parameters were measured among 627, 386, 362, and 628 participants, respectively. We found that sperm MMP was significantly positively correlated with all of conventional semen parameters including semen volume (r = 0.090, p = 0.025), sperm concentration (r = 0.301, p<0.01), total sperm count (r = 0.324, p<0.01), and progressive motility (r = 0.399, p<0.01); sperm MMP was also negatively correlated with Annexin V+ sperm (r = -0.553, p<0.01); mtDNAcn was significantly negatively correlated with sperm concentration (r = -0.214, p<0.01), total sperm count (r = -0.232, p<0.01), and progressive motility (r = -0.164, p = 0.01); mtDNA integrity was also significantly positively correlated with sperm concentration (r = 0.195, p<0.01), total sperm count (r = 0.185, p<0.01), and progressive motility (r = 0.106, p = 0.043). After adjusting for potential confounders, these relationships remained significant. Furthermore, we explored the potential effects of lifestyles on such mitochondrial biomarkers and found that the current drinkers displayed a higher level of sperm MMP; additionally, mt DNAcn was increased with age. The results indicated that certain mitochondrial biomarkers could serve as predictors of semen quality in a general population, and the study provides a baseline for the effects of population characteristics and lifestyles on such mitochondrial markers.

9 EVALUATION OF A MODIFIED GnRH-BASED TIMED ARTIFICIAL INSEMINATION PROTOCOL ASSOCIATED WITH ESTRUS DETECTION IN CYCLIC BEEF HEIFERS INSEMINATED WITH SEX-SELECTED SEMEN

This study compared pregnancy per TAI (P/TAI) between sex-selected and conventional semen in cyclic beef heifers subjected to the modified (without initial GnRH) 5-day Co-synch protocol and evaluated the usefulness of an oestrus detection (ED) to identify heifers that were most likely to conceive. Heifers (n = 1034; 13 to 15 months of age) at three locations were scanned by ultrasonography to determine cyclicity (presence of corpus luteum) and reproductive tract normalcy. Cyclic heifers (n = 940) received a 1.38-g progesterone-releasing device (CIDR; Zoetis Animal Health, Florham Park, NJ, USA) on Day 0, CIDR removal and 500 μg of cloprostenol (Estroplan; Vetoquinol NA Inc., Edmonton, AB, Canada) on Day 5, and 100 μg of GnRH (Fertiline; Vetoquinol NA Inc.) along with TAI on Day 8 (72 ± 1 h after CIDR removal). On Day 5, heifers received ED patches (EstrotectTM; Estrotect Inc.) that were scored from 0 to 3, based on colour change between initial application and Day 8; 0 = unchanged, 1 = ≤50% colour change, 2 = &gt;50% colour change, 3 = missing. Heifers were assigned by ear-tag number to be inseminated with frozen–thawed sex-selected or conventional semen from either of two sires available commercially. Pregnancy diagnosis was performed by transrectal ultrasonography 28 (herd A) or 48 (herds B and C) days after TAI. Data were analysed using the PROC GLIMMIX in SAS (SAS Institute Inc., Cary, NC, USA), with herd included as a random effect. The CIDR retention rate was 98%; P/TAI in heifers that lost the CIDR was 21%. Pregnancy per TAI [% (no.)] did not differ (P &gt; 0.05) between sires [58.4 (426) v. 58.0 (414) for sires A and B, respectively], but it was higher (P &lt; 0.01) for conventional semen [63.7 (422) v. 52.6 (418)]. An interaction between sire and type of semen (P = 0.07) revealed that sex-selected semen from bull B had the lowest P/TAI [49.3 (203)]. Overall, heifers with an ED patch scored 2 had greater (P &lt; 0.01) P/TAI [64.8 (526)] than those scored 0 [42.9 (170)] or 1 [46.8 (79)]. Heifers with a missing ED patch had intermediate P/TAI [58.5 (65)]. However, the effect of ED patch scoring on P/TAI depended on type of semen. In heifers inseminated with conventional semen, PTAI was significantly lower (P &lt; 0.01) in those with an ED patch scored 0 [49.4(87)] compared to those with a patch scored 2 [69.6 (270)]. In heifers inseminated with sex-selected semen, P/TAI was significantly lower (P &lt; 0.05) in those with an ED patch scored 0 [36.1(83)] or 1 [37.2 (43)] compared with those scored 2 [59.8 (256)] or 3 [58.3 (36)]. In summary, P/TAI was greater in heifers inseminated with conventional semen. However, the P/TAI obtained with the modified GnRH-based TAI protocol in cyclic heifers inseminated with sex-selected semen was encouraging. In addition, the Estrotect oestrus detection patches were considered useful to identify animals for TAI with sex-selected semen and could be used to increase the adoption of this technology in beef herds. This study was supported by Alberta Agriculture and Forestry, Vetoquinol NA Inc., Estrotect Inc. and collaborative beef producers.

190 SexedULTRA™, A NEW METHOD OF PROCESSING SEX-SORTED BOVINE SPERM IMPROVES CONCEPTION RATES

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 &lt; 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 &lt; 0.001) 56-day nonreturn rate followed by conventional 15.0 (66.73 v. 65.66%); XY 2.1 presented the lowest (P &lt; 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

Proteomic analysis of seminal plasma and sperm cells and their associations with semen freezability in Guzerat bulls.

The objective of the present study was to describe the relationship of seminal plasma and total sperm cell proteins with the semen freezability parameters of Guzerat bulls. Thirteen bulls were subjected to breeding soundness evaluation. Semen samples were collected, cryopreserved, and then post-thawing sperm kinetics were assessed, where high ( = 7) and low ( = 6) freezability groups were defined. Seminal plasma and total sperm proteins from the 2 groups were separated by 2-dimensional SDS-PAGE, and spots were identified by mass spectrometry. Semen parameters post-cryopreservation were as follows in the high and low freezability groups, respectively: mean total motility, 52.4 ± 20.5 and 13.7 ± 3.9; percentage of normal sperm, 89.0 ± 2.6 and 64.7 ± 14.0; and reactivity of hypo-osmotic swelling test, 38.9 ± 4.7 and 13.6 ± 3.7. Three seminal plasma proteins (osteopontin-K, DNase γ precursor, and DNASE1L3) and 6 proteins from sperm cells (acrosome formation-associated factor isoform 2, annexin A1, disintegrin and metalloproteinase domain-containing protein 2, dihydrolipoyl dehydrogenase, and glyceraldehyde-3-phosphate dehydrogenase) were highly expressed ( < 0.05) in the high freezability group. Another 6 seminal plasma proteins (acrosin inhibitor 1, glutathione peroxidase 3, metalloproteinase inhibitor 2, ephrin-A1, annexin A1, and platelet-activating factor acetylhydrolase) were significantly higher ( < 0.05) in the low freezability group. We described the associations of seminal plasma and sperm cell proteins with post-thawing sperm viability of Guzerat bulls raised in a typical semiarid environment. Such associations indicate that specific seminal plasma proteins more abundant in bulls of low semen freezability may be a response to an early oxidative stress that is not detected by conventional prefreezing semen evaluation. Moreover, specific sperm proteins were more associated with good freezability. The results presented here can serve as guidelines for future research aiming to develop better extenders and/or to improve bull semen selection for cryopreservation.

Coenzyme Q10 Intake From Food and Semen Parameters in a Subfertile Population

To assess the association between coenzyme Q10 (CoQ10) intake from food sources and semen quality. We assessed this association in a prospective cohort of men attending a fertility clinic. CoQ10 supplementation has been associated with improvements in semen parameters. However, the impact of CoQ10 intake from food sources on semen quality has not been investigated. Subfertile couples seeking fertility evaluation at the Massachusetts General Hospital Fertility Center were invited to participate in an ongoing study of environmental factors and fertility. In total, 211 male participants completed a validated food frequency questionnaire and provided 476 semen samples. Multivariable linear mixed models were used to examine the relation between CoQ10 intake from food and semen parameters while adjusting for potential confounders and accounting for within-person correlations. Mean dietary CoQ10 intake was 19.2 mg/day (2.4-247.2 mg/day). No subjects were taking CoQ10 supplements. There were no associations between dietary CoQ10 intake from food and conventional semen parameters. The adjusted mean difference (95% confidence interval) comparing men in the top and bottom quartiles of CoQ10 intake from food were -3.1 mil/mL (95% confidence interval -29.5, 38.8 mil/mL) for sperm concentration, -4.5% (-15.1%, 6.0%) for total motility, -1.3% for progressive motility (-8.4%, 5.7%), and 0.3% (-1.4%, 2.0%) for sperm morphology. CoQ10 intake from food was not related to semen parameters among subfertile men. Mean dietary intake of CoQ10 in this study was 10-fold lower than the supplemental dose used in clinical trials showing improved sperm motility. CoQ10 intake from food alone may be insufficient to optimize semen parameters.

Mitochondrial Functionality and Chemical Compound Action on Sperm Function.

During the last decade, several studies have shown that mitochondrial parameters, such as integrity, respiratory activity, membrane potential and ROS production are intimately linked with sperm quality. Given the limitations of conventional semen analyses in terms of predicting male fertility, an increasing number of studies are focusing on the characterization of sperm mitochondria in order to more accurately assess sperm functionality. Moreover, mitochondria from several organs, such as the liver, have been described as a powerful screening tool for drug safety, being an easy in vitro model to assess the toxicity of distinct families of compounds. Given that mitochondrial functionality is intimately related to sperm homeostasis, it has become important to understand how compounds, ranging from dietary supplements, environmental pollutants, dependency-inducing drugs to pharmacological agents (such as erectile dysfunction-targeted drugs and male contraceptives) affect sperm mitochondrial function. In this review, we discuss studies describing the effects of various chemical agents on spermatozoa, with particular emphasis on mitochondrial function. From the extensive literature analyzed, we conclude that in some cases the role of sperm mitochondria as putative predictors of sperm functionality is very obvious, while in others further studies are needed to clarify this issue.

Evaluation of spermatozoal function-useful tools or just science.

Conventional microscopic semen analysis does not provide precise information on the fertilizing potential of a male. The traditional basis for semen evaluation is that male fertility is dependent on production of a "proper" concentration/number of motile, morphologically normal spermatozoa in excess to achieve conception. Many independent studies, especially in human medicine, have demonstrated that the absolute number of spermatozoa does not accurately determine fertility, but their functional competence. Many functional tests of spermatozoa are developed over the last decades. Computer-assisted sperm analysis (CASA) and flow cytometry have become the gold standard for semen assessment in specialized andrology laboratories. Other functional assays, such as gamete interaction tests, provide additional information regarding the real fertilizing potential of sperm cells. From this point of view, such tests are valuable diagnostic tools in fertility disorders and may be helpful to make a decision which method of treatment to use: pharmacological therapy, intrauterine insemination, introduction of classic IVF, ICSI or exclusion from a breeding programme. The most useful gamete interaction tests include induced acrosome reaction, zona pellucida binding assay, oocyte penetration assay and hyaluronan binding assay. In recent years, andrology has entered into a new era of sophisticated OMICS methods. Genomics, epigenomics, transcriptomics and proteomics brought high hopes for rapid progress in clinical diagnostics.

Quality and developmental rate of embryos produced with sex-sorted and conventional semen from superovulated dairy cattle

This study investigated the effect of sex-sorted semen compared with conventional semen on the outcome of embryo recovery, placing special emphasis on the quality, and developmental stage of embryos. Data were analyzed for 443 embryo collections with sex-sorted semen (SEX group) and 1528 with conventional semen (CONV group) in superovulated dairy heifers and cows. The insemination protocol for conventional semen included two inseminations, comprising a total dose of 30 million sperm passing into the uterine body. For sex-sorted semen, two (30%) to three (70%) deep uterine inseminations were performed, the total dose ranging from eight to 12 million sperm. The data were analyzed separately for heifers and cows. The total number of recovered structures was similar among the groups. The number of viable embryos decreased in the SEX groups compared with the CONV (with 1.4 and 3.2 fewer embryos in heifers and cows, correspondingly, P < 0.001), and correspondingly the proportions of unfertilized ova and degenerated embryos increased in the SEX groups (P < 0.001). The proportion of unsuccessful collections, yielding no transferable embryos, increased in the SEX groups for both heifers (from 7.2% to 11.2%, P = 0.025) and cows (from 9.0% to 20.7%, P < 0.001). Regarding the quality of viable embryos, the quality grades were superior in the CONV group compared with the SEX group for heifers (P < 0.001) and cows (P < 0.001). The proportion of grade 1 embryos decreased by 6.5 percentage points in heifers and 11.9 percentage points in cows when sex-sorted semen was used. Correspondingly, the proportions of grade 2 and 3 embryos increased in heifers and cows when sexed semen was used. The mean developmental stages of embryo collections were numerically slightly lower in the SEX group. In heifers, the delay in developmental stage was statistically significant (P = 0.001), but in cows, there was only a tendency toward that (P = 0.067). In conclusion, sex-sorted sperm decreased the transferable embryo yield and increased the risk of a recovery yielding no transferable embryos. Furthermore, use of sex-sorted semen decreased the proportion of grade 1 embryos. In addition, it also seemed to delay embryonic development, although the delay in embryonic development was minimal and its biological relevance remains undefined. Despite the compromised embryo production, taken into account the optimization of recipient resources, the use of sex-sorted semen is advantageous, especially in superovulated heifers, which are of most importance in the modern breeding strategies using genomic selection.

Are oxidative stress markers associated with unexplained male infertility?

Male infertility can be responsible for up to 20% of the cases attending fertility consultation facilities; nonetheless, the underlying molecular mechanisms that could explain it are still elusive. Therefore, we aimed to evaluate conventional and functional parameters of semen samples from patients who presented with male infertility of unknown origin. Conventional semen parameters and functional parameters (i.e. intracellular reactive oxygen species production, mitochondrial membrane potential, sperm chromatin structure assay, sperm membrane lipid peroxidation and antioxidant capacity of seminal plasma) were evaluated on semen samples from 54 healthy donors, 23 patients with idiopathic infertility and 34 fertile controls. No significant differences were observed in the conventional seminal parameters between the fertile and infertile men. However, increased intracellular reactive oxygen species (ROS) production and DNA fragmentation were observed in the infertile patients compared to the fertile group. Alterations in intracellular ROS production and DNA fragmentation could be associated with male idiopathic infertility. These parameters could eventually distinguish both groups more accurately than the conventional parameters. Our current results are encouraging, and the efficacy of these parameters in the clinical settings needs to be further assessed to establish their predictive potential as a marker of unexplained male infertility.

Stochastic dynamic simulation modeling including multitrait genetics to estimate genetic, technical, and financial consequences of dairy farm reproduction and selection strategies.

The objective of this study was to develop a daily stochastic dynamic dairy simulation model that included multitrait genetics and to evaluate the effects of reduced genetic models and various reproduction and selection strategies on the genetic, technical, and financial performance of a dairy herd. The 12 correlated genetic traits included in the 2014 lifetime net merit (NM$) index were modeled for each animal. For each animal, a true breeding value (TBV) for each trait was calculated as the average of the sire's and dam's TBV, plus a fraction of the inbreeding and Mendelian sampling variability. Similarly, an environmental component for each trait was calculated and was partitioned into a permanent and a daily (temporary) effect. The combined TBV and environmental effects were converted into the phenotypic performance of each animal. Hence, genetics and phenotypic performances were associated. Estimated breeding values (EBV) were also simulated. Genetic trends for each trait for the service sire were based on expected trends in US Holsteins. Surplus heifers were culled based on various ranking criteria to maintain a herd size of 1,000 milking cows. In the first 8 scenarios, culling of surplus heifers was either random or based on the EBV of NM$. Four different genetic models, depending on the presence or absence of genetic trends or genetic and environmental correlations, or both, were evaluated to measure the effect of excluding multitrait genetics on animal performance. In the last 5 scenarios, the full genetic model was used and culling of surplus heifers was either random or based on the EBV of NM$ or the EBV of milk. Sexed semen use and reliability of the EBV were also varied. Each scenario was simulated for 15yr into the future. Results showed that genetic models without all 12 genetic trends and genetic and environmental correlations provided biased estimates of the genetic, technical, and financial performance of the dairy herd. Average TBV of NM$ of all cows in the herd was $263 greater in yr 15 in a scenario that combined sexed semen use in heifers and culling of surplus heifers with the lowest EBV of NM$, compared with a scenario that used only conventional semen and surplus heifers were culled randomly. The average TBV of daughter pregnancy rate of all cows in the herd was 1.25 percentage points greater in yr 15 in a scenario that combined using sexed semen in heifers as well as culling of surplus heifers ranked by EBV of NM$, compared with a scenario using conventional semen only as well as culling surplus heifers ranked by EBV of milk. In conclusion, the multitrait genetics model resulted in improved estimates of genetic, technical, and financial effects and appears useful to evaluate consequences of various reproduction and selection strategies within a dairy farm.

Modern Reproductive Technologies to Improve Cattle Production

Cattle contribute significantly to the global supply of animal-derived proteins which are an important part of a well-balanced human diet. With the human population increasing by 1 billion people every 13 years, there will be an estimated 9.6 billion people on planet Earth by the year 2050. To attain global food security the amount of available food will need to double between now and then. Enhancing reproductive efficiency is a prerequisite for boosting production of meat and milk from cattle, and, fortunately, an arsenal of modern reproductive technologies is available to assist with that effort. The objective of this manuscript is to provide an overview of reproductive biotechnologies that can bolster the efficient production of meat and milk from cattle. Protocols for synchronization of estrus and synchronization of ovulation facilitate more efficient artificial insemination using conventional or sex-sorted semen. In vivo and in vitro production of preimplantation embryos from genetically superior females enable creation of multiple offspring with high production potential. Biochemical and ultrasonographic methods for pregnancy testing identify non-pregnant females that can be re-mated or sold to prevent wastage of valuable feed resources. Somatic cell nuclear transfer is used to create copies of highly productive animals, and genome editing of zygotes provides a novel opportunity to selectively enhance the genetic makeup of cattle for the benefit of animal and human health. Wise use of these reproductive technologies will increase food production from cattle and will help alleviate world hunger.