Determination of enzyme activity in rabbit seminal plasma and its relationship with quality semen parameters

M.P. Viudes de Castro, L. Casares-Crespo, A. Monserrat-Martínez, J.S. Vicente


The objective of this study was to determine rabbit seminal plasma enzyme activity. Furthermore, correlations between semen parameters and enzyme activity and male age were examined. The study was performed using 17 New Zealand White males from 5 to 9 mo old. Overall, 252 semen samples were collected from bucks from May to September. Semen characteristics were analysed and the seminal plasma was obtained by centrifugation. The activities of alanyl aminopeptidase (APN), aspartate transaminase (AST), alanine aminotransferase (ALT), γ-glutamyl transpeptidase (GGT), lactate dehydrogenase (LDH) and alkaline phosphatase (ALKP) in the seminal plasma fluid were measured. Significant differences between males were found in APN, GGT, LDH, ALKP and ALT activities (P<0.05). No significant differences between enzyme activity and male age were found. We also observed significant positive correlations between male age and sperm concentration (r=0.26), progressive motility (r=0.17) and amplitude of lateral head displacement (r=0.21), and negative ones between male age and average path velocity (r=–0.56), velocity of the sperm head along its actual curvilinear path (r=–0.61), straight line velocity (r=–0.50), linearity index (r=–0.13), and cytoplasmic droplet (r=–0.33). Furthermore, a significant negative correlation between APN activity and the status of the acrosome (r=–0.20) and significant positive correlations between APN activity and the sperm abnormalities (r=0.21), GGT activity and sperm concentration (r=0.34) and the status of the acrosome (r=0.31), and ALKP activity and sperm concentration were observed (r=0.41). In our study, APN and GGT seem to be the most predictive enzymes for rabbit semen quality.


rabbit; seminal plasma; enzyme activity; seminal parameters

Full Text:



Alvariño J.M.R., 2000. Reproductive performance of male rabbits. In Proc.: 7th World Rabbit Congress, July 4-7, Valencia, Spain, 2308-1910.

Agrawal Y., Vanha-Perttula T. 1986. Alanyl aminopeptidase of bovine seminal vesicle secretion. Int. J. Biochem., 18: 725-729. doi:10.1016/0020-711X(86)90396-4

Attia Y.A., Kamel K.I. 2012. Semen quality, testosterone, seminal plasma biochemical and antioxidant profiles of rabbit bucks fed diets supplemented with different concentrations of soybean lecithin. Animal, 6: 824-833. doi:10.1017/S1751731111002229

Cheng L., Lin Y.L., Peng G., Li F. 2012. Structural basis for multifunctional roles of mammalian aminopeptidase N. Proc. Natl. Acad. Sci. USA, 109:17966-17971. doi:10.1073/pnas.1210123109

Dogan I., Polat U., Nur Z. 2009. Correlations between seminal plasma enzyme activities and semen parameters in seminal fluid of Arabian horses. Iran. J. Vet. Res., Shiraz University, 10: Ser.No.27.

Farrell P.B., Foote R.H., Simkin M.E., Clegg E.D., Wall R.J. 1993. Relationship of semen quality, number of sperm inseminated, and fertility in rabbits. J. Androl., 14: 464-471.

Fernández D., Valdivia A., Irazusta J., Ochoa C., Casis L. 2002. Peptidase activities in human semen. Peptides, 23: 461-468. doi:10.1016/S0196-9781(01)00622-2

Huang K., Takahara S., Kinouchi T., Takayama M., Ishida T., Ueyama H., Nishi K., Ohkubo I. 1997. Alanyl aminopeptidase from human seminal plasma: purification, characterization and immunohistochemical localization in the male genital tract. J. Biochem., 122: 779-787. doi:10.1093/oxfordjournals.jbchem.a021823

Irazusta J., Valdivia A., Fernández D., Agirregoiia E., Ochoa C., Casis L. 2004. Enkephalin-degrading enzymes in normal and subfertile human semen. J. Androl., 25: 733-739.

Kiso W.K., Selvaraj V., Nagashima J., Asano A., Brown J.L., Schmitt D.L., Leszyk J., Travis A.J., Pukazhenthi B.S. 2013. Lactotransferrin in Asian Elephant (Elephas maximus) seminal plasma correlates with semen quality. PLOS One, 8: e71033. doi:10.1371/journal.pone.0071033

Kozdrowski R., Dubiel A. 2004. The effect of season on the properties of wild boar (Sus scrofa L.) semen. Anim. Reprod. Sci., 80: 281-289. doi:10.1016/j.anireprosci.2003.08.006

La Falci V.S., Tortorella H., Rodrigues J.L., Brandelli A. 2002. Seasonal variation of goat seminal plasma proteins. Theriogenology, 57: 1035-1048. doi:10.1016/S0093-691X(01)00714-2

Lavara R., Mocé E., Lavara F., Viudes-de-Castro M.P., Vicente J.S. 2005. Do parameters of seminal quality correlate with the results of on-farm inseminations in rabbits? Theriogenology, 64: 1130-1141. doi:10.1016/j.theriogenology.2005.01.009

Luzi F., Maertens L., Lijten P., Pizzi F. 1996. Effect of feeding level and dietary protein content on libido and semen characteristics of bucks. In Proc.: 6th World Rabbit Congress, July 9-12, 1996, Toulouse, France, vol. 2, 87-92.

Mann T., Lutwak-Mann C. 1981. Biochemistry of spermatozoa: chemical and functional correlations in ejaculated semen. In: Male Reproductive Function and Semen. Themes and Trends in Physiological and Biochemical and Investigative Andrology, ed. T. Mann and Lutwak-Mann, Spring-Verlag: Berlin, 1981, p. 195-268.

Miros V.V., Mikhno V.I. 1982. Semen quality of male rabbits in relation to age and season. Inst. Zhivotnovodstva Lesotepi i Polesya, 34: 45-48.

Muller B. 1983. Genital tract proteins in the male rabbit: Alkaline phosphatase enzyme action and site of synthesis. Andrologia, 15: 676-681. doi:10.1111/j.1439-0272.1983.tb00191.x

Okab A.B. 2007. Semen characteristics and plasma testosterone of New-Zealand male rabbits are affected by environmental temperatures. Slovak J. Anim. Sci. 40: 161-167.

Osada T., Watanabe G., Kondo S., Toyoda M., Sakaki Y., Takeuchi T. 2001. Male reproductive defects caused by puromycinsensitive aminopeptidase deficiency in mice. Mol. Endocrinol., 15: 960-971. doi:10.1210/mend.15.6.0643

Pesch S., Bergmann M., Bostedt H. 2006. Determination of some enzymes and macro- and microelements in stallion seminal plasma and their correlations to semen quality. Theriogenology, 66: 307-313. doi:10.1016/j.theriogenology.2005.11.015

Salzberger Z., Lewin L.M., Shalgi R. 1992. Loss of acid phosphatase from rat spermatozoa as a method for assessing the acrosome reaction. Andrologia, 24: 155-159. doi:10.1111/j.1439-0272.1992.tb02630.x

Sirat M.P., Sinha A.K., Singh B.K., Prasad R.L. 1996. Effect of cryoprotectants on release of various enzymes from buck spermatozoa during freezing. Theriogenology, 45: 405-416. doi:10.1016/0093-691X(95)00377-K

Smith P.K., Krohn R.I., Hermanson G.T., Mallia A.K., Gartner F.H., Provenzano M., Fujimoto E.K., Goeke N.M., Olson B.J., Klenk D.C. 1985. Measurement of protein using bicinchoninic acid. Analyt. Bioch., 150: 76-5. doi:10.1016/0003-2697(85)90442-7

Theau-Clément M., Bolet G., Sanchez A., Saleil G., Brun J.M. 2015. Some factors that influence semen characteristics in rabbits. Anim Reprod. Sci., 157: 33-38. doi:10.1016/j.anireprosci.2015.03.011

Urner F., Sakkas D. 2003. Protein phosphorylation in mammalian spermatozoa. Reproduction, 125: 17-26. doi:10.1530/rep.0.1250017

Viudes-de-Castro M.P., Mocé E., Lavara R., Marco-Jiménez F., Vicente J.S. 2014. Aminopeptidase activity in seminal plasma and effect of dilution rate on rabbit reproductive performance after insemination with an extender supplemented with buserelin acetate. Theriogenology, 81: 1223-1228. doi:10.1016/j.theriogenology.2014.02.003

Xu X., Pommier S., Arbov T., Hutchings B., Sotto W., Foxcroft G.R. 1998. In vitro maturation and fertilization techniques for assessment of semen quality and boar fertility. J. Anim. Sci., 76: 3079-3089.

Yousef M.I. 2005. Reproductive performance, blood testosterone, lipid peroxidation and seminal plasma biochemistry of rabbits as affected by feeding Acacia saligna under subtropical conditions. Food Chem. Toxicol., 43: 333-339. doi:10.1016/j.fct.2004.10.006

Yousef M.I., Abdallah G.A., Kamel K.I. 2003. Effect of ascorbic acid and Vitamin E supplementation on semen quality and biochemical parameters of male rabbits. Anim. Reprod. Sci., 76: 99-111. doi:10.1016/S0378-4320(02)00226-9

Yousef M.I., Esmail A.M., Baghdadi H.H. 2004. Effect of isoflavones on reproductive performance, testosterone levels, lipid peroxidation and seminal plasma biochemistry of male rabbits. J. Environ. Sci. Heal. B, 39: 819-833. doi:10.1081/LESB-200030880

Yousef M.I., El-Demerdash F.M., Kamil K.I., Elaswad F.A.M. 2006. Ameliorating effect of folic acid on chromium(VI)-induced changes in reproductive performance and seminal plasma biochemistry in male rabbits. Reprod. Toxicol., 21: 322-328. doi:10.1016/j.reprotox.2005.09.005

Yousef M.I., Kamel K.I., Hassan M.S., El-Morsy A.M.A. 2010. Protective role of propolis against reproductive toxicity of triphenyltin in male rabbits. Food Chem. Toxicol., 48: 1846-1852. doi:10.1016/j.fct.2010.04.018

Abstract Views

Metrics Loading ...

Metrics powered by PLOS ALM


Cited-By (articles included in Crossref)

This journal is a Crossref Cited-by Linking member. This list shows the references that citing the article automatically, if there are. For more information about the system please visit Crossref site

1. Reproductive characteristics, semen quality, seminal oxidative status, steroid hormones, sperm production efficiency of rabbits fed herbal supplements
Olatunji Abubakar Jimoh, Wahab Adekunle Oyeyemi, Hafsat Ololade Okin-Aminu, Bolaji Fatai Oyeyemi
Theriogenology  vol: 168  first page: 41  year: 2021  
doi: 10.1016/j.theriogenology.2021.03.020


 Universitat Politècnica de València


Official journal of the World Rabbit Science Association (WRSA)


e-ISSN: 1989-8886     ISSN: 1257-5011