Hendricks K.E.M.,University of Florida |
Penfold L.M.,White Oak Conservation Center |
Evenson D.P.,SCSA Diagnostics |
Kaproth M.T.,Genex Cooperative Inc. |
And 2 more authors.
Theriogenology | Year: 2010
Biological samples, including cryopreserved sperm, are routinely X-rayed during air shipment. The goal was to investigate the impact of X-irradiation used for checked and carry-on luggage on bovine sperm chromatin integrity and postfertilization in vitro embryonic development. Frozen domestic bull sperm (Bos taurus) (n = 9 bulls) stored in a dry shipper (-160 °C) was screened by X-irradiation 0, 1, 2, and 3 times as either carry-on or checked luggage. Duplicate straws were thawed, and sperm were assessed for chromatin damage using the sperm chromatin structure assay (SCSA) and by postfertilization in vitro developmental competence of mature oocytes. Multiple exposure to X-rays did not significantly affect sperm chromatin integrity assessed by SCSA. There were lower proportions of oocytes cleaved (P = 0.07; 21.6 ± 3.1% vs. 29.4 ± 3.1%, 24.9 ± 3.1%, and 25.7 ± 3.3% for 3 vs. 0, 1, and 2 times, respectively; least-squares means ± SEM) and that developed to blastocysts (P = 0.06; 9.0 ± 1.7% vs. 13.8 ± 1.7%, 11.5 ± 1.7%, and 12.6 ± 1.9%, respectively) when fertilization was performed with sperm X-rayed 3 times using checked luggage irradiation; developmental competence (percentage cleaved embryos becoming blastocysts) was unaffected. There were no deleterious effects of other X-irradiation treatments on embryo development. We inferred that screening by X-irradiation may reduce the ability of sperm to activate oocyte cleavage after multiple exposures at the checked luggage dose. However, there was no evidence that competence of embryos to become blastocysts was reduced by X-irradiation (45.4 ± 5.7%, 40.4 ± 5.7%, 46.4 ± 6.1%, and 41.8 ± 5.7% for 0, 1, 2, and 3 doses, respectively), but potential long-term epigenetic effects are unknown. © 2010.
Card C.J.,University of Rhode Island |
Anderson E.J.,University of Rhode Island |
Zamberlan S.,University of Rhode Island |
Krieger K.E.,Genex Cooperative Inc. |
And 2 more authors.
Biology of Reproduction | Year: 2013
Ejaculated bovine spermatozoa retain a pool of RNAs that may have a function in early embryogenesis and be used as predictors of male fertility. The bovine spermatozoal transcript profile remains incomplete because previous studies have relied on hybridization-based techniques, which evaluate a limited pool of transcripts and cannot identify full-length transcripts. The goal of this study was to sequence the complete cryopreserved bovine spermatozoal transcript profile using Illumina RNA-Sequencing (RNA-Seq). Spermatozoal RNA was pooled from nine bulls with conception rate scores ranging from -2.9 to 3.5 and confirmed to exclude genomic DNA and somatic cell mRNA. After selective amplification of poly(A)+ RNA and high-throughput sequencing, 6166 transcripts were identified via alignment to the bovine genome (UMD 3.1/bosTau6). RNA-Seq transcript levels (n = 9) were highly correlated with quantitative PCR copy number (r2 = 0.9747). The bovine spermatozoal transcript profile is a heterogeneous population of degraded and full-length predominantly nuclear-encoded mRNAs. Highly abundant spermatozoal transcripts included PRM1, HMGB4, and mitochondrial-encoded transcripts. Full-length transcripts comprised 66% of the top 368 transcripts (fragments per kilobase of exon per million fragments mapped [FPKM] . 100) and amplification of the full-length transcript or 5 ′and 3 ′ends was confirmed for selected transcripts. In addition to the identification of transcripts not previously reported in spermatozoa, several known spermatozoal transcripts from various species were also found. Gene ontology analysis of the FPKM . 100 spermatozoal transcripts revealed that translation was the most predominant biological process represented. This is the first report of the spermatozoal transcript profile in any species using high-throughput sequencing, supporting the presence of mRNA in spermatozoa for further functional and fertility studies. © 2013 by the Society for the Study of Reproduction, Inc.
PubMed | University of Rhode Island and Genex Cooperative Inc.
Type: | Journal: Animal reproduction science | Year: 2017
Spermatozoal messenger RNA (mRNA) has the potential as a molecular marker for sire fertility because this population can reflect gene expression that occurred during spermatogenesis and may have a functional role in early embryonic development. The goal of this study was to compare the oligo-dT selected spermatozoal transcript profiles of higher fertility (Conception Rate (CR) 1.8-3.5) and lower fertility (CR -2.9 to -0.4) sires using Ribonucleic Acid Sequencing (RNA-Seq). A total of 3227 transcripts and 5366 transcripts were identified in the higher and lower fertility populations, respectively. While common transcripts between the two populations were identified (2422 transcripts), several transcripts were also unique to the fertility populations including 805 transcripts that were unique to the higher fertility population and 2944 transcripts that were unique to the lower fertility population. From gene ontological analysis, the transcripts unique to each fertility population differed in Biological Processes (BP), including enrichment of regulatory transcripts for growth and protein kinase activity in the higher fertility bulls. Biological variation in transcript presence among individual sires was also found. Of the candidate fertility spermatozoal transcripts chosen from the RNA-Seq population analysis reported here and previous publications, COX7C was negatively correlated with sire fertility. Using high-throughput sequencing, candidate spermatozoal transcripts were identified for further study as potential markers for sire fertility.
Mallory D.A.,University of Missouri |
Lock S.L.,Genex Cooperative Inc. |
Woods D.C.,Genex Cooperative Inc. |
Poock S.E.,University of Missouri |
Patterson D.J.,University of Missouri
Journal of Dairy Science | Year: 2013
The objective was to compare pregnancy per AI (P/AI) with conventional (CON) or sex-sorted (SS) semen from a single sire within a fixed-time AI (FTAI) program designed for dairy heifers. Holstein heifers (n=240) were assigned to treatment (CON or SS) according to body weight and reproductive tract score. All heifers underwent FTAI by using the " Show-Me-Synch" protocol [controlled internal drug release (CIDR) insert from d 0 to 14 followed by PGF2α (25mg i.m.) 16d after insert removal (d 30) with GnRH (100μg i.m.) and FTAI at 66h after PGF2α]. A single professional technician performed the FTAI. Heifers were fitted with heat detection patches at PGF2α to characterize estrous response. Estrous response did not differ between CON (63/120; 53%) and SS (70/120; 58%) treatments. The CON heifers, however, achieved greater FTAI P/AI (82/120; 68%) compared with SS (45/120; 38%) heifers. The P/AI did not differ for CON heifers that exhibited or failed to exhibit estrus before FTAI [44/63 (70%) vs. 38/57(67%), respectively]. For SS heifers, however, those that exhibited estrus had greater P/AI compared with those that failed to exhibit estrus [32/70 (46%) vs. 13/50 (26%)]. Pregnancy per AI resulting from FTAI was greater for heifers that were inseminated with CON semen compared with those that received SS semen. The expression of estrus before FTAI did not affect P/AI when CON semen was used, whereas the P/AI with SS semen was greater for heifers detected in estrus. Further studies are required to develop strategies for using sex-sorted semen when inseminating heifers at predetermined fixed times on the basis of expression of estrus before FTAI. © 2013 American Dairy Science Association.
Thomas J.M.,University of Missouri |
Lock S.L.,Genex Cooperative Inc. |
Poock S.E.,University of Missouri |
Ellersieck M.R.,University of Missouri |
And 2 more authors.
Journal of Animal Science | Year: 2014
This experiment was designed to test the hypothesis that delayed insemination of nonestrous cows would increase pregnancy rates when using sexsorted semen in conjunction with fixed-time artificial insemination (FTAI). Estrus was synchronized for 656 suckled beef cows with the 7-d CO-Synch + controlled internal drug release (CIDR) protocol (100 μg GnRH + CIDR [1.38 g progesterone] on d 0, 25 mg PGF2α at CIDR removal on d 7, and 100 μg GnRH on d 10, 66 h after CIDR removal). Estrus detection aids (Estrotect) were applied at PGF2α and CIDR removal on d 7, and estrous expression was recorded at GnRH on d 10. Cows were assigned to 1 of 3 treatments: 1) FTAI (concurrent with GnRH, 66 h after CIDR removal) with conventional semen regardless of estrous expression, 2) FTAI with sex-sorted semen regardless of estrous expression, or 3) FTAI with sex-sorted semen for cows having expressed estrus and delayed AI 20 h after final GnRH for cows failing to express estrus. A treatment × estrous expression interaction was found (P < 0.0001). Higher pregnancy rates (P < 0.0001) were achieved with conventional semen (Treatment 1; 77%) than with sex-sorted semen (Treatments 2 and 3; 51 and 42%, respectively) among cows that expressed estrus. However, among cows that failed to express estrus, delayed insemination with sex-sorted semen yielded higher (P < 0.0001) pregnancy rates than with sexsorted semen at the standard time (Treatments 2 and 3; 3 versus 36%, respectively). Furthermore, among cows that failed to express estrus, FTAI pregnancy rates when using sex-sorted semen at the delayed time (36%) were comparable (P = 0.9) to those achieved using conventional semen at the standard time (Treatment 1; 37%). These results indicate that delaying AI of nonestrous cows by 20 h from the standard FTAI improves pregnancy rates when sex-sorted semen is used with FTAI. © 2014 American Society of Animal Science. All rights reserved.
Kennedy C.E.,University of Missouri |
Krieger K.B.,Genex Cooperative Inc. |
Sutovsky M.,University of Missouri |
Xu W.,MOFA ICB |
And 7 more authors.
Molecular Reproduction and Development | Year: 2014
SUMMARY: Post-acrosomal WW-domain binding protein (PAWP) is a signaling molecule located in the post-acrosomal sheath (PAS) of mammalian spermatozoa. We hypothesized that the proper integration of PAWP in the sperm PAS is reflective of bull-sperm quality and fertility. Cryopreserved semen samples from 298 sires of acceptable, but varied, fertility used in artificial insemination services were analyzed using immunofluorescence microscopy and flow cytometry for PAWP protein. In normal spermatozoa, PAWP fluorescence formed a regular band around the proximal PAS. Anomalies of PAWP labeling in defective spermatozoa were reflected in flow cytometry by varied intensities of PAWP-induced fluorescence. Distinct sperm phenotypes were also identified, including morphologically normal and some defective spermatozoa with moderate levels of PAWP; grossly defective spermatozoa with low/no PAWP; and defective spermatozoa with high PAWP. Analysis by ImageStream flow cytometry confirmed the prevalence of abnormal sperm phenotypes in the spermatozoa with abnormal PAWP content. Live/dead staining and video recording showed that some abnormal spermatozoa are viable and capable of progressive motility. Conventional flow-cytometric measurements of PAWP correlated significantly with semen quality and fertility parameters that reflect the sires' artificial insemination fertility, including secondary sperm morphology, conception rate, non-return rate, and residual value. A multiplex, flow-cytometric test detecting PAWP, aggresomes (ubiquitinated protein aggregates), and acrosomal integrity (peanut-agglutinin-lectin labeling) had a predictive value for conception rate, as demonstrated by step-wise regression analysis. We conclude that PAWP correlates with semen/fertility parameters used in the cattle artificial insemination industry, making PAWP a potential biomarker of bull fertility. © 2014 Wiley Periodicals, Inc.
Abdel-Azim G.,Genex Cooperative Inc.
Journal of Dairy Science | Year: 2010
Field data were collected over a period of 2 yr by artificial insemination technicians for the purpose of evaluating differences among bulls in their fertility when synchronization and semen sorting were involved. First, main effects of synchronization and semen sorting were found to reduce bull fertility by 1.5 and 12.7%, respectively. Second, the interaction of both factors with bull fertility significantly enhanced the evaluation models. Differences between 2 sets of adjusted conception rates for synchronized and nonsynchronized services ranged from 0.5 to 2.9%, whereas differences between 2 sets of adjusted conception rates for sorted and conventional semen ranged from -1.8 to 15.2%. This implies that using conventional fertility models that ignore these effects may not be sufficiently accurate in situations where synchronization or semen sorting are involved. Accounting for synchronization and especially for semen sorting to evaluate bulls on their fertility and the production of separate sets of conception rates under each situation are essential. © 2010 American Dairy Science Association.
Bjelland D.W.,University of Wisconsin - Madison |
Weigel K.A.,University of Wisconsin - Madison |
Coburn A.D.,Genex Cooperative Inc. |
Wilson R.D.,Genex Cooperative Inc.
Journal of Dairy Science | Year: 2015
Recent evidence has suggested that some of the decline in reproductive ability in dairy cattle has been caused by embryonic death. The current study compared expected genomic inbreeding from sire-dam mating pairs to genomic inbreeding from live progeny in an attempt to determine how embryonic inbreeding may affect fertility. A total of 11,484 Holstein cattle with 43,485 SNP markers and pedigree information were available for analysis. A total of 412 sire-dam-progeny trios in which all animals had reliable genotypes were discovered. After removal of trios because of parentage errors, 374 remained for analysis. Additionally, a total of 3,031 animals comprising 3,906 genotyped full-sibling pairs were available for comparison. Expected genomic inbreeding measures were calculated by predicting homozygosity independently per SNP (FPHE) in sire-dam mating pairs and by simulating progeny using phased haplotype information (FROHE and FPHE). Actual genomic inbreeding measures were calculated using the percent homozygosity of all SNP (FPH) and using runs of homozygosity (FROH). Average FPHE values (62.8±0.78%) were slightly lower than FPH (63.1±1.12%), when considering each SNP independently. After phasing haplotypes, FPHE (62.5±0.83%) was again slightly lower than FPH (62.7±1.16%), and FROHE (3.46±1.54%) was slightly lower than FROH (3.53±2.17%). Results suggest increases in expected genomic inbreeding do not explain a large effect on embryo viability at average levels of expected inbreeding. Higher variation in FROH values was present with sire-dam mating pairs exhibiting high FROHE, which may suggest high levels of genomic inbreeding are required for a noticeable effect on overall embryo viability. Genomic inbreeding between full siblings was also compared with moderate correlations (0.47-0.52) present. Overall, expected genomic inbreeding measures were calculated, but results did not suggest a large effect of expected inbreeding on embryo viability. © 2015 American Dairy Science Association.
PubMed | Genex Cooperative Inc.
Type: Journal Article | Journal: Journal of dairy science | Year: 2010
Field data were collected over a period of 2 yr by artificial insemination technicians for the purpose of evaluating differences among bulls in their fertility when synchronization and semen sorting were involved. First, main effects of synchronization and semen sorting were found to reduce bull fertility by 1.5 and 12.7%, respectively. Second, the interaction of both factors with bull fertility significantly enhanced the evaluation models. Differences between 2 sets of adjusted conception rates for synchronized and nonsynchronized services ranged from 0.5 to 2.9%, whereas differences between 2 sets of adjusted conception rates for sorted and conventional semen ranged from -1.8 to 15.2%. This implies that using conventional fertility models that ignore these effects may not be sufficiently accurate in situations where synchronization or semen sorting are involved. Accounting for synchronization and especially for semen sorting to evaluate bulls on their fertility and the production of separate sets of conception rates under each situation are essential.