Animal Reproduction (AR)
Animal Reproduction (AR)

Evaluation of novel SexedULTRA-4M technology for in vitro bovine embryo production

Horacio Álvarez-Gallardo; Michael Edward Kjelland; Mario Pérez-Martínez; Fernando Villaseñor-González; Salvador Romo-García

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SexedULTRA-4M™ is made using an improved method of sex-sorting sperm in a less damaging environment for better retaining sperm integrity throughout the sorting process. The objective of this research was to compare conventional (CONV) and SexedULTRA-4M™ (ULTRA-4M) semen for bovine IVP using four Angus bulls. Matured slaughterhouse oocytes (n = 4000) were divided into the CONV group and the ULTRA-4M group (2000 COCs for each semen type). The IVF process was implemented with CONV and ULTRA-4M semen from the same bull. The cleavage rates, eight cell embryos and blastocysts on day 7 of culture were evaluated for each semen type and each bull. The statistical analysis was carried out with the ANOVA procedure SAS software. The results were 54.45% ± 1.03 and 58.10% ± 1.07; 35% ± 1.57 and 39.15% ± 1.62; 22.8% ± 1.09 and 27.15% ± 1.12 for CONV and ULTRA-4M, respectively, for cleavage rate, eight cell embryos and blastocysts on day 7 for the average of all bulls, comparing only the semen type. Concerning only the semen type, ULTRA-4M was significantly superior to CONV for cleavage rates (P = 0.01) and blastocysts on day 7 (P = 0.009). There were no significant differences between the CONV and ULTRA-4M groups (P>0.05) for all variables analyzed for Bull 1 and Bull 4, however, for Bull 2 ULTRA-4M was significantly superior to CONV for cleavage rates and blastocysts on day 7 (P< 0.05). In Bull 3, ULTRA-4M was significantly higher (P< 0.05) for blastocysts on day 7 compared to CONV. In conclusion, under the conditions of this research the ULTRA-4M and CONV semen produced similar bovine IVP results overall.


SexedULTRA-4M, sexed semen, bovine, IVF


Aguila L, Treulen F, Therrien J, Felmer R, Valdivia M, Smith LC. Oocyte selection for in vitro embryo production in bovine species: noninvasive approaches for new challenges of oocyte competence. Animals. 2020;10(12):2196. PMid:33255250.

Barceló-Fimbres M, Campos-Chillón LF, Seidel GE Jr. In vitro fertilization using non-sexed and sexed bovine sperm: sperm concentration, sorter pressure, and bull effects. Reprod Domest Anim. 2011;46(3):495-502. PMid:20946538.

Cran DG, Johnson LA, Miller NG, Cochrane D, Polge C. Production of bovine calves following separation of X- and Y-chromosome bearing sperm and in vitro fertilisation. Vet Rec. 1993;132(2):40-1. PMid:8442336.

Cran DG, Johnson LA, Polge C. Sex preselection in cattle: a field trial. Vet Rec. 1995;136(19):495-6. PMid:7645188.

Ferré LB, Chitwood JL, Fresno C, Ortega HH, Kjelland ME, Ross PJ. Effect of different mini-volume colloid centrifugation configurations on flow cytometrically sorted sperm recovery efficiency and quality using a computer-assisted semen analyzer. Reprod Domest Anim. 2018;53(1):26-33. PMid:28891229.

Fischer-Brown A, Barquero G, Clark S, Ferguson C, Ireland F, Jensen N, Lane S, Lindsey B, Lopes P, Monson R, Northey D, Reeder A, Rutledge J, Wheeler M, Kesler D. Twin vs. single transfer of IVP Holstein embryos to beef recipients. Reprod Fertil Dev. 2005;17(2):230.

Folchini NP, Leivas FG, Santos FW, Schwengber EB, Martin DM, Spiazzi CC, Brum DS. Uso de mini-Percoll modificado para seleção e redução da formação de espécies reativas de oxigênio (ROS) em espermatozoides bovinos. Rev Bras Reprod Anim. 2002;36(4):239-44.

González-Marín C, Góngora CE, Gilligan TB, Evans KM, Moreno JF, Vishwanath R. In vitro sperm quality and DNA integrity of SexedULTRA sex-sorted sperm compared to non-sorted bovine sperm. Theriogenology. 2018;114:40-5. PMid:29597122.

Gonzalez-Marin C, Lenz RW, Gilligan TB, Evans KM, Gongora CE, Moreno JF, Vishwanath R. SexedULTRA™, a new method of processing sex-sorted bovine sperm improves post-thaw sperm quality and in vitro fertility. Reprod Fertil Dev. 2017;29(1):204.

Lenz RW, Gonzalez-Marin C, Gilligan TB, DeJarnette JM, Utt MD, Helser LA, Hasenpusch E, Evans KM, Moreno JF, Vishwanath R. SexedULTRA™, a new method of processing sex-sorted bovine sperm improves conception rates. Reprod Fertil Dev. 2017;29(1):203-4.

Lu KH, Cran DG, Seidel GE Jr. In vitro fertilization with flow-cytometrically-sorted bovine sperm. Theriogenology. 1999;52(8):1393-405. PMid:10735084.

Lu KH, Seidel GE Jr. Effects of heparin and sperm concentration on cleavage and blastocyst development rates of bovine oocytes inseminated with flow cytometrically-sorted sperm. Theriogenology. 2004;62(5):819-30. PMid:15251233.

Machado GM, Carvalho JO, Filho ES, Caixeta ES, Franco MM, Rumpf R, Dode MA. Effect of Percoll volume, duration and force of centrifugation, on in vitro production and sex ratio of bovine embryos. Theriogenology. 2009;71(8):1289-97. PMid:19230963.

Merton JS, Haring RM, Stap J, Hoebe RA, Aten JA. Effect of flow cytometrically sorted frozen thawed semen on success rates of in vitro bovine embryo production. Theriogenology. 1997;47(1):295.

Missio D, Folchini NP, Leivas FG, Pavin CIIUM, Pinto HF, Cibin FWS, Brum DDS. Reduction in Percoll volume increases recovery rate of sex-sorted semen of bulls without affecting sperm quality and early embryonic development. Anim Reprod Sci. 2018;192:146-53. PMid:29534826.

Mostek A, Janta A, Ciereszko A. Proteomic comparison of non-sexed and sexed (X-bearing) cryopreserved bull semen. Anim Reprod Sci. 2020;221:106552. PMid:32861114.

Oliveira LZ, Lima VFH, Levenhagen MA, Santos RM, Assumpção TI, Jacomini JO, Andrade AF, Arruda RP, Beletti ME. Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa. J Vet Sci. 2011;12(3):267-72. PMid:21897100.

Palma GA, Olivier NS, Neumüller C, Sinowatz F. Effects of sex-sorted spermatozoa on the efficiency of in vitro fertilization and ultrastructure of in vitro produced bovine blastocysts. Anat Histol Embryol. 2008;37(1):67-73. PMid:18197903.

Seidel GE Jr. Economics of selecting for sex: the most important genetic trait. Theriogenology. 2003;59(2):585-98. PMid:12499006.

Seidel GE Jr. Update on sexed semen technology in cattle. Animal. 2014;8(Suppl 1):160-4. PMid:24680061.

Seidel GE Jr, Garner DL. Current status of sexing mammalian spermatozoa. Reproduction. 2002;124(6):733-43. PMid:12537000.

Simões R, Feitosa WB, Siqueira AF, Nichi M, Paula-Lopes FF, Marques MG, Peres MA, Barnabe VH, Visintin JA, Assumpção ME. Influence of bovine sperm DNA fragmentation and oxidative stress on early embryo in vitro development outcome. Reproduction. 2013;146(5):433-41. PMid:23940385.

Steele H, Makri D, Maalouf WE, Reese S, Kölle S. Bovine Sperm Sexing Alters Sperm Morphokinetics and Subsequent Early Embryonic Development. Sci Rep. 2020;10(1):6255. PMid:32277124.

Thomas JM, Locke JWC, Bonacker RC, Knickmeyer ER, Wilson DJ, Vishwanath R, Arnett AM, Smith MF, Patterson DJ. Evaluation of SexedULTRA 4M™ sex-sorted semen in timed artificial insemination programs for mature beef cows. Theriogenology. 2019;123:100-7. PMid:30296650.

Thomas JM, Locke JWC, Vishwanath R, Hall JB, Ellersieck MR, Smith MF, Patterson DJ. Effective use of SexedULTRA™ sex-sorted semen for timed artificial insemination of beef heifers. Theriogenology. 2017;98:88-93. PMid:28601161.

Vishwanath R, Moreno JF. Review: semen sexing - current state of the art with emphasis on bovine species. Animal. 2018;12(s1):s85-96. PMid:29552998.

Wheeler MB, Rutledge JJ, Fischer-Brown A, VanEtten T, Malusky S, Beebe DJ. Application of sexed semen technology to in vitro embryo production in cattle. Theriogenology. 2006;65(1):219-27. PMid:16263159.

Wilson RD, Fricke PM, Leibfried-Rutledge ML, Rutledge JJ, Penfield CM, Weigel KA. In vitro production of bovine embryos using sex-sorted sperm. Theriogenology. 2006;65(6):1007-15. PMid:16122781.

Wilson RD, Weigel KA, Fricke PM, Rutledge JJ, Leibfried-Rutledge ML, Matthews DL, Schutzkus VR. In vitro production of Holstein embryos using sex-sorted sperm and oocytes from selected cull cows. J Dairy Sci. 2005;88(2):776-82. PMid:15653544.

Zhang M, Lu KH, Seidel GE Jr. Development of bovine embryos after in vitro fertilization of oocytes with flow cytometrically sorted, stained and unsorted sperm from different bulls. Theriogenology. 2003;60(9):1657-63. PMid:14580648.

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