Sperm characteristics, metabolic markers.


This study investigated the effect of different sperm freezability on some semen characteristics and amino acid concentrations in the seminal plasma of Holstein bulls cryopreserved for two years. The semen straws (0.25 ml) that cryopreserved for two years were divided into three equal groups (7 bulls per group) according to the sperm’s freezability: medium (73.02%; F1), good (83.76%; F2), and high (91.20%; F3). The F1 bulls were superior (P≤ 0.05) in sperm’s cell individual motility (SCIM) for fresh semen as compared with F2 and F3 bulls. The F2 and F3 groups were superior (P≤ 0.01) to the F1 group in live sperm percentage at cryopreservation. The sperm’s DNA damage percentage was lesser (P≤ 0.05) in the F1 and F3 groups as compared with the F2 group. The differences among groups in normal sperm percentage, sperm’s plasma membrane and acrosome integrity percentage, malondialdehyde, and total antioxidant concentrations lacked significance. The essential and non-essential amino acid concentrations in seminal plasma did not differ among groups. In conclusion, sperm freezability influenced some semen attributes but did not affect amino acid concentrations in the seminal plasma of Holstein bulls.


Ahmed, S., M. I. U. Khan, M. Ahmed and S. Iqbal. 2017. Effect of age on lipid peroxidation of fresh and frozen-thawed semen of Nili-Ravi buffalo bulls. Italian J. Anim. Sci., 17 (3), 730-735

AL-Gebouri, F.G. and. S.M. Eidan. 2023. Metabolites and semen characteristics in different bulls fertility. Iraqi Journal of Agricultural Sciences, 55(Special Issue), 206-216. https://doi.org/10.36103/ijas.v55iSpecial.1899.

Al-Helal, A. M. and T. A. Abdulkareem. 2023. Effect of adding resveratrol to soybean-lecithin extender on semen attributes of buffalo bulls. Iraqi Journal of Agricultural Sciences, 54 (4): 1074-1083. https://doi.org/10.36103/ijas.v54i4.1797

Ali, M. D., F. F. Ibrahim, T. A. Abdulkareem, S. M. Eidan, Q. A. Ameen, M. G. Abdul-Karim, and O. A. Mohammed. 2015. Evaluation of Artificial Insemination Activities for Cattle in Iraq: Survey Study. Ministry of Agriculture, pp. 48-51

Al-Nuaimi, A. J. J. 2019. Effect of adding Olea europaea and Rosmarinus officinalis aqueous extracts and calcium to Tris and Soybean-Lecithin extenders on activating cooled and post-cryopreserved low-semen quality of Holstein bulls. Ph. D. Dissertation. College of Agricultural Engineering Sciences, University of Baghdad. pp: 86

Al-Nuaimi, A. J., T. A. Abdulkareem, F. F. Ibrahim, Z. A. Humade and F. A. Hussein. 2020. effect of adding Olea europaea and Rosmarinus officinalis aqueous extracts and calcium chloride to Tris extender on post-cryopreserved sperm’s cell individual motility and live sperm percentage for low semen quality of Holstein bulls. ‏ Biochem. Cell. Arch., 20 (1), 519-524. https://doi.org/10.35124/bca.2020.20.1.519

Al-Saedy, A. J. A. and T. A. Abdulkareem. 2022. Comparison of semen quality for three lines of Holstein bulls. 1. Some immediate and microscopic characteristics. Iraqi Journal of Agricultural Sciences, 53 (4), 752-759. https://doi.org/10.36103/ijas.v53i4.1585

Alwaeli, S. N. and S.M. Eidan.2024. Synergistic effect of sperm filtration technique and antioxidants added to extender of Iraqi buffalo semen. Iraqi Journal of Agricultural Sciences, 55(1), 413-421. https://doi.org/10.36103/kc809146.

Alwaeli, S.N. and Eidan, S.M. 2023. Effect of Glass wool and Sephadex sperm separation techniques on improving the poor quality semen of Iraqi buffalo bulls (4th–ICMTAS) IOP Conference Series: Earth and Environmental Science 1262(7), 072003. https://doi.org/10.1088/1755-315/1262/7/072003

Bailey, J., A. Morrier and N. Cormier. 2003. Semen cryopreservation: successes and persistent problems in farm species. Canadian Journal of Animal Science, 83: 393-401. https://doi.org/10.4141/A03-02

Bieniek, J. M., A. P. Drabovich and K. Lo. 2016. Seminal biomarkers for the evaluation of male infertility. Asian J. Androl., 18(3),426–433. https://doi.org/10.4103/1008-682X.175781

Birben, E., U.M. Sahiner, C. Sackesen, S. Erzurum and O. Kalayci. 2012. Oxidative stress and antioxidant defense. World Allergy Organ. J., 5(1), 9-19.

DOI: 10.1097/WOX.0b013e3182439613

Bustani, G. S and Baiee, F. H. 2021. Semen extenders: An evaluative overview of preservative mechanisms of semen and semen extenders. Veterinary World, 14(5), 1220-1233. https://doi.org/10.14202/vetworld.2021.1220-1233

Cheng, Y., X. Yang, X. Deng, X. Zhang, P. Li, J. Tao, C. Qin, J. Wei and Q. Lu. 2015. Metabolomics in bladder cancer: a systematic review. Int. J. Clin. Exp. Med. 8, 11052–11063

Eidan, S. M.2016. Effect on post-cryopreserved semen characteristics of Holstein bulls of adding combinations of vitamin C and either catalase or reduced glutathione to Tris extender. Anim. Reprod. Sci., 167, 1-7. https://doi.org/10.1016/j.anireprosci.2016.01.014

Eidan, S.M., O.A.A. Sultan, F.F. Ibrahim, K.F. Hurish, and H.H. Naseer. 2017a. Effect of adding Manganese chloride and Co-enzymes (α-lipoic acid and Q10) on post-cryopreservation semen quality characteristics of Holstein bulls. Iraqi J. Vet. Med., 41(2), 86-93. https://doi.org/10.30539/iraqijvm.v41i2.53

Eidan, S. M., O. H. Al-Zaidi, F. F. Ibrahim, B. A. R. Al-Timimi, and W. E. Lateef. 2015. Effect of adding catalase and glutathione reduce to Tris extender on freezing ability of Holstein bulls following different cryopreservation periods. The Iraqi J. Vet. Med., 39 (2), 19-24. https://doi.org/10.30539/iraqijvm.v39i2.161

Hitit, M., M. R. Ugur, T. T. N. Dinh, D. Sajeev, A. Kaya, E. Topper, W. Tan and E. Mimili. 2020. Cellular and functional physiopathology of bull sperm with altered sperm freezability. Front. Vet. Sci., 7, Article # 581137. https://doi.org/10.3389/fvets.2020.581137

Kumar D, P. Kumar, P. Singh, SP. Yadav, SK. Sarkar, A. Bharadwaj and PS. Yadav. 2014. Characteristics of frozen-thawed semen in predicting the fertility of buffalo bulls. Indian J. Anim. Sci., 84, 389–92. https://doi.org/10.56093/ijans.v84i4.39838

Kumaresan, A., A. Johannisson, E. M. Al-Essawe and J. M. Morrell.2017. Sperm viability, reactive oxygen species, and DNA fragmentation index combined can discriminate between above-and below-average fertility bulls. J. Dairy Sci., 100 (7), 5824-5836. https://doi.org/10.3168/jds.2016-12484

Kundu, C. N., K. Das and G. C. Majumder. 2001. Effect of amino acids on goat cauda epididymal sperm cryopreservation using a chemically defined model system. Cryobiology, 42 (1),21-27. https://doi.org10.1006/cryo.2001.2296

Leushuis, E., J. W. Van Der Steeg, P. Steures, S. Repping, P. M. Bossuyt, M. A. Blankenstein and P. G. Hompes. 2010. Reproducibility and reliability of repeated semen analyses in male partners of subfertile couples. Fertil. Steril., 94(7), 2631-2635. https://doi.org/10.1016/j.fertnstert.2010.03.021

Lone, S.A., T.K. Mohanty, R.K. Baithalu and H.P. Yadav. 2019. Sperm protein carbonylation. Andrologia, 51, e13233. https://doi.org/10.1111/and.13233

Mahmood, N. M., S.M. Eidan, R. I. Khalil, F. F. Ibrahim, W.Y. Lateef and T. A. Shihab. 2019. Effect of melatonin hormone on long-term cryopreserved semen: 2-characteristics of semen Holstein bulls. Biochem. Cell. Arch., 19 (1), 1429-1434. https://doi.org/10.35124/bca.2019.19.1.1429

Mohammad Abadi, F., A. Mirfazeli, H. Zaeri, M. Nejabat, M. Ariaie and H. Joshaghani. 2016. Analysis of plasma amino acids using RP-HPLC and pre-column derivatization with OPA/3-MPA. Medical Laboratory Journal, 10(2), 52-57

Mostek, A., M.A. Dietrich, M. Słowi´nska and A. Ciereszko. 2017. Cryopreservation of bull semen is associated with carbonylation of sperm proteins. Theriogenology, 92, 95–102. https://doi.org/10.1016/j.theriogenology.2017.01.011

Musa, K.S., and T.A. Abdulkareem. 2023. Protein profiles in seminal plasma of Iraqi buffalo bulls (Bubalus bubalis) associated with fresh and cryopreserved semen quality. IOP Conf. Series: Earth and Environmental Science 1262 (2023), 072095. https://doi.org/10.1088/1755-1315/1262/7/072095

Musa, K.S., and T.A. Abdulkareem. 2024a. Regression coefficient of fresh and cryopreserved semen attributes of Iraqi buffalo bulls (Bubalus bubalis) on protein types in seminal plasma. IOP Conf. Series: Earth and Environmental Science 1302 (1), 012055. https://doi.org/10.1088/1755-1315/1302/1/012055

Oberoi, B., S. Kumar, and P. Talwar. 2014. Study of human sperm motility post cryopreservation. Med. J. Armed. Forces. India 70, 349–353. https://doi.org/10.1016/j.mjafi.2014.09.006

Recuero, S., B. Fernandez-Fuertes, S. Bonet, L. Barranco and M. Yeste. 2019. Potential of seminal plasma to improve the fertility of frozen-thawed boar spermatozoa. Theriogenology, 137, 36–42. https://doi.org/10.1016/j.theriogenology.2019.05.035

Sharma, R., G. Ahmad, S. C. Esteves, and A. Agarwal. 2016. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay using bench top flow cytometer for evaluation of sperm DNA fragmentation in fertility laboratories: protocol, reference values, and quality control. J. Assist. Reprod. Genet., 33(2), 291-300. https://doi.org/10.1007/s10815-015-0635-7

SAS. 2012. Statistical Analysis System, User's Guide. Statistical. Version 9.1th ed. SAS. Inst. Inc. Cary. N.C. USA

Souza, A. L. P., G. L. Lima, G. C. X. Peixoto, A. M. Silva, M. F. Oliveira and A. R. Silva. 2016. Use of Aloe vera–based extender for chilling and freezing collared peccary (Pecari tajacu) semen. Theriogenology, 85(8), 1432-1438. https://doi.org/10.1016/j.theriogenology.2016.01.007

Sultan, O. A. A. 2018. Relationship of osteopontin, CD9 and COX-2 genes with semen characteristics of Holstein bulls. Ph.D. Dissertation, College of Agricultural Engineering Sciences, University of Baghdad

Ugur, M. R., A. S. Abdelrahman, H. C. Evans, A. A. Gilmore, M. Hitit, R. I. Ariflantini, B. Purwantara, A, Kaya and E. Memili. 2019. Advances in cryopreservation of bull sperm: A review. Front. Vet. Sci., 6, Article # 268. https://doi.org/10.3389/fvets.2019.00268

Ugur, M. R., T. Dinh, M. Hitit, A. Kaya, E. Topper, B. Didion and E. Memili. 2020. Amino acids of seminal plasma associated with the freezability of bull sperm. Front. Cell Dev. Biol., 7, Article #347. https;//doi.org/10.3389/fcell.2019.00347

Velho, A. L. C., E. Menezes, T. Dinh, A. Kaya, E. Topper, A. A. Moura and E. Memili. 2018. Metabolomic markers of fertility in bull seminal plasma. PLoS One, 13(4), e0195279. https://doi.org/10.1371/journal.pone.0195279

Zuidema, D., K. Kern, and P. Sutovsky. 2021. An exploration of current and perspective semen analysis and sperm selection for livestock artificial insemination. Animals, 11, Article # 3563. https://doi.org/10.3390/ani11123563







How to Cite


Publication Dates

Similar Articles

1-10 of 213

You may also start an advanced similarity search for this article.