Animal Reproduction (AR)
Animal Reproduction (AR)

Spirulina platensis extract addition to semen extender enhances cryotolerance and fertilizing potentials of buffalo bull spermatozoa

Magdy Badr; Zaher Rawash; Ahmed Azab; Ragab Dohreg; Taha Ghattas; Mohamed Fathi

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The objective of this study was to investigate the effect of Spirulina platensis extract (SPE) addition to the freezing extender on freezability, lipid peroxidation, ultrastructure alterations and fertilizing potentials of frozen-thawed buffalo bull spermatozoa. Semen samples were collected with artificial vagina from five adult fertile bulls and diluted with Tris-base extender containing SPE (1, 5, 10 and 20 μg/mL) or without SPE (control). Diluted semen was cooled to 4 °C throughout one hour and frozen in 0.25 mL straws: prior to being stored in liquid nitrogen. Cryopresreved spermatozoa were assessed for post-thawing sperm motility, viability, acrosomal integrity, ultrastructure changes, antioxidant activities, lipid peroxidation and fertility rate. The current results clearly indicated that adding 10μg/mL SPE to the freezing extender significantly improved (P< 0.05) post-thawing motility and decrease the percentage of acrosomal damage (51.67±6.02% and 16.33±1.46%, respectively) compared with the control (28.33±4.41% and 26.33±1.77%, respectively). Moreover, addition of 10 μg/mL SPE to the semen extender significantly diminished (P< 0.05) MDA concentration (10.66±2.40 nmol/109) compared with the control (22.66±4.26 nmol/109). Therefore, the present results revealed that addition of 10μgl/mL SPE to the freezing extender might improve semen quality and reduce cryodamage of the buffalo bull spermatozoa.


spirulina, buffalo bull, spermatozoa, cryopreservation, lipid peroxidation


Abd El-Baky HH, El-Baz FK, El-Baroty GS. Enhancement of antioxidant production in Spirulina platensis under oxidative stress. American- Eurasian J. Sci. Res. 2007;2:170-9.

Abshenas J, Babaei H, Zare MH, Allahbakshi A, Sharififar F. The effect of green tea (Camellia sinensis) extract on mouse semen quality after scrotal heat stress. Vet Res Forum. 2011;4:242-7.

Aitken RJ, Ryan AL, Baker MA, McLaughlin EA. Redox activity associated with the maturation and capacitation of mammalian spermatozoa. Free Radic Biol Med. 2004;36(8):994-1010. PMid:15059640.

Aitken RJ. Reactive oxygen species as mediators of sperm capacitation and pathological damage. Mol Reprod Dev. 2017;84(10):1039-52. PMid:28749007.

Alvarez JG, Storey BT. Spontaneous lipid peroxidation in rabbit epididymal spermatozoa: its effect on sperm motility. Biol Reprod. 1982;27(5):1102-8. PMid:6297627.

Antonio Marcos EG, Monroy Hermosillo O, Cortes Barberena E, Rodriguez Tobon E, Rivera Martinez JG, Arenas Rios E. Can C-phycocyanin be used as a sperm protector? Veterinaria Mexico OA. 2018;5(4):1-9.

Asghari A, Fazilati M, Latifi AM, Salavati H, Choopani A. Antioxidant properties of Spirulina. J Appl Biotechnol Rep. 2016;3:345-51.

Azevedo HC. (2006). Spermatic integrity and function in RAM cryopreserved sêmen after incorporation of cholesterol, desmosterol, oleic–linoleic acid and -lactalbumin [thesis]. Botucatu: Faculty of Veterinary Medicine and Animal Science, UNESP.

Babadzhanov AS, Abdusamatova N, Yusupova FM, Faizullaeva N, Mezhlumyan LG, Malikova MK. Chemical composition of Spirulina platensis cultivated in Uzbekistan. Chem Nat Compd. 2004;40(3):276-9.

Badr MR, El-Malak MJA, Hassan HM. Effect of trehalose on cryopreservation, oxidative stress and DNA integrity of buffalo spermatozoa. J Reprod Infertil. 2010;2:50-7.

Baruselli PS, Carvalho NAT, Gimenes LU, Crepaldi GA. Fixed-time artificial insemination in buffalo. Ital J Anim Sci. 2007;6:107-18.

Bashandy S, Mohamed M, Fatma A. Spirulina platensis, reduced liver and kidney injuries induced by sodium arsenite. Int J Pharm Tech Res. 2016;11:35-48.

Bilodeau JF, Blanchette S, Gagnon C, Sirard MA. Thiols prevent H O - mediated loss of sperm motility in cryopresreved bull semen. Theriogenology. 2001;56(2):275-86. PMid:11480619.

Boonkusol D, Saikhun K, Ratanaphumma P. Effect of extender and storage time on motility and ultrastructure of cooled-preserved boar spermatozoa. Witthayasan Kasetsat Witthayasat. 2010;44:582-9.

Bucak MN, Sariozkan S, Tuncer PB, Ulutas PA, Akcadag HAA. Effect of antioxidants on microscopic semen parameter, lipidperoxidation and antioxidant activities in Angora goat semen following cryopreservation. Small Rumin Res. 2009;81(2-3):90-5.

Calamera JC, Fernandez PJ, Buffone MG, Acosta AA, Doncel GF. Effects of long-term in vitro incubation of human spermatozoa: functional parameters and catalase effect. Andrologia. 2001;33(2):79-86. PMid:11350371.

Chu W, Lim Y, Radhakrishnan AK, Lim PE. Protective effect of aqueous extract from Spirulina platensis against cell death induced by free radicals. BMC Complement Altern Med. 2010;10(1):53. PMid:20858231.

Cingi C, Conk-Dalay M, Cakli H, Bal C. The efects of Spirulina on allergic rhinitis. Eur Arch Otorhinolaryngol. 2008;265(10):1219-23. PMid:18343939.

Djalil OIA, Ngoula Ferdinand N, Adoum IY, Tchoffo Herve T, Djalal AK, Vemo BN, Herman NV. Effects of hydro-ethanolic extract of Spirulina (Spirulina platensis) on semen characteristics and oxidative stress indicators in male rabbit (Oryctolagus cuniculus). J Vet Med Animal Sci. 2019;2(1):1010.

El-Amrawi GA. Study on capacitation of buffalo spermatozoa. In: Egyptian Society Animal Reproduction Fertility 3rd Annual Congress; 1992; Cairo. Cairo: ESARF; 1992. p. 214-22.

El-Ratel IT, Gabr AA. Potential impact of Spirulina Alga as an antioxidant on improving semen production and oxidative stress in blood and seminal plasma of rabbit bucks. Egypt Poult Sci J. 2020;40(1):209-24.

El-Shahat KH, Taysser MI, Badr MR, Zaki K. Investigating the effects of theophylline on motility, hyperactivity, and acrosome reaction of spermatozoa in rams. Veterinary Medicine and Public Health Journal. 2020;3:81-4.

El-Tantawy WH. Antioxidant effects of Spirulina supplement against lead acetate-induced hepatic injury in rats. J Tradit Complement Med. 2016;6(4):327-31. PMid:27774414.

El-Tohamy MM, El-Nattat WS. Effect of antioxidant on lead-induced oxidative damage and reproductive dysfunction in male rabbits. J Am Sci. 2010;6:613-22.

Esener OBB, Gurel-Gurevin E, Isbilen-Basok B, Yigit F, Bilal T, Altiner A, Yilmazer N, Armutak EI. Spirulina platensis affects factors involved in spermatogenesis and increases ghrelin receptors in testis tissue of rats fed a high-fat diet. Pol J Vet Sci. 2017;20(3):467-75. PMid:29166288.

Farag MR, Alagawany M, Abd El-Hac ME, Dhama K. Nutritional and healthical aspects of Spirulina (Arthrospira) for poultry, animals and human. Int J Pharmacol. 2016;12(1):36-51.

Farber JL, Kyle ME, Coleman JB. Mechanisms of cell injury by activated oxygen species. Lab Invest. 1990;62(6):670-9. PMid:2162996.

Flohé L, Otting F. Superoxide dismutase assays. Methods Enzymol. 1984;105:93-104. PMid:6328209.

Ford WCL. Regulation of sperm function by reactive oxygen species. Hum Reprod Update. 2004;10(5):387-99. PMid:15218008.

Fouda S, Ismail R. Effect of Spirulina Platensis on reproductive performance of rabbit bucks. Egypt J Nutr Feeds. 2017;20(1):55-66.

Granaci V. Achievements in the artificial insemination of swine. Bull Univ Agric Sci Vet Med Cluj-Napoca Anim Sci Biotechnol. 2007a;63/64:382-6.

Granaci V. Contributions on the study of the cryoresistance increase of the bull semen material. Bull Univ Agric Sci Vet Med Cluj-Napoca Anim Sci Biotechnol. 2007b;63/64:387-91.

Gupta S, Hrishikeshvan HJ, Sehajpal PK. Spirulina protects against rosiglitazone induced osteoporosis in insulin resistance rats. Diabetes Res Clin Pract. 2010;87(1):38-43. PMid:19896232.

Gutiérrez-Rebolledo GA, Galar-Martínez M, Garcia-Rodriguez RV, Chamorro-Cevallos GA, Hernández-Reyes AG, Martinez-Galero E. Antioxidant effect of Spirulina (Arthrospira) maxima on chronic inflammation induced by Freund’s complete adjuvant in rats. J Med Food. 2015;18(8):865-71. PMid:25599112.

Hoque MN, Talukdar AK, Kamal M, Kumar AJ, Farida YB, Shamsuddin M. Ovulation synchronization in water buffaloes guided by milk progesterone Elisa. J Embryo Transf. 2011;26:105-9.

Hosseini SM, Khosravi-Darani K, Mozafari MR. Nutritional and medical applications of Spirulina microalgae. Mini Rev Med Chem. 2013;13(8):1231-7. PMid:23544470.

Joyce DA. Oxygen radicals in disease. Adverse Drug React Bull. 1987;127(1):476-9.

Keservani RK, Kesharwani RK, Sharma AK, Jarouliya U. Dietary supplements, nutraceutical, and functional foods in immune response (immunomodulators). In: Bagchi D, Preuss HG, Swaroop A, editors. Nutraceutical and functional foods in human life and disease prevention. Boca Raton: CRC Press, Taylor and Francis; 2015. chap. 20, p. 343-60.

Khan Z, Bhadouria P, Bisen PS. Nutritional and therapeutic potential of Spirulina. Curr Pharm Biotechnol. 2005;6(5):373-9. PMid:16248810.

Kistanova E, Marchev Y, Nedeva R, Kacheva D, Shumkov K, Georgiev B, Shimkus A. Effect of the Spirulina platensis included in the main diet on the boar sperm quality. Biotechnol Anim Husb. 2009;25(5-6-1):547-57.

Kumar P, Saini M, Kumar D, Bharadwaj A, Yadav PS. Estimation of endogenous levels of osteopontin, total antioxidant capacity and malondialdehyde in seminal plasma: application for fertility assessment in buffalo (Bubalus bubalis) bulls. Reprod Domest Anim. 2017;52(2):221-6. PMid:27862407.

Kurd F, Samavati V. Water soluble polysaccharides from Spirulina platensis: extraction and in vitro anti-cancer activity. Int J Biol Macromol. 2015;74:498-506. PMid:25583023.

Leboeuf B, Restall B, Salamon S. Production and storage of goat semen for artificial insemination. Anim Reprod Sci. 2000;62(1-3):113-41. PMid:10924822.

Medeiros CMO, Forell F, Oliveira ATD, Rodrigues JL. Current status of sperm cryopreservation: why isn’t it better? Theriogenology. 2002;57(1):327-44. PMid:11775978.

Mendes RL, Nobre BP, Cardoso MT, Pereira AP, Palavra AF. Supercritical carbon dioxide extraction of compounds with pharmaceutical importance from microalgae. Inorg Chim Acta. 2003;356:328-34.

Mizera A, Kuczaj M, Szul A. Impact of the Spirulina maxima extract addition to semen extender on bovine sperm quality. Ital J Anim Sci. 2019;18(1):601-7.

Nah WH, Koh IK, Ahn HS, Kim MJ, Kang H-G, Jun JH, Gye MC. Effect of Spirulina maxima on spermatogenesis and steroidogenesis in streptozotocin-induced type I diabetic male rats. Food Chem. 2012;134(1):173-9.

Neild DM, Gadella BM, Chaves MG, Miragaya MH, Colenbrander B, Agüero A. Membrane changes during different stages of a freeze-thaw protocol for equine semen cryopreservation. Theriogenology. 2003;59(8):1693-705. PMid:12566145.

Paul V, Prakash BS. Efficacy of the ovsynch protocol for synchronization of ovulation and fixed-time artificial insemination in Murrah buffaloes (Bubalus bubalis). Theriogenology. 2005;64(5):1049-60. PMid:16125551.

Placer ZA, Cushman LL, Johnson BC. Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Anal Biochem. 1966;16(2):359-64. PMid:6007581.

Rahman MA, Moitry NF, Alam M, Yasmin Z, Debnath D, Mostofa M. Effects of Spirulina in lead induced toxicities in long evans rats. J Environ Sci Nat Resour. 2012;5(1):79-82.

Sedlak J, Lindsay RHC. Estimation of total, protein-bound and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem. 1968;25(1):192-205. PMid:4973948.

Sikka SC, Rajasekaran M, Hellstrom WJ. Role of oxidative stress and antioxidants in male infertility. J Androl. 1995;16(6):464-8. PMid:8867595.

Storey B. Mammalian sperm metabolism:oxygen and sugar, friend and foe. Int J Dev Biol. 2008;52(5-6):427-37. PMid:18649255.

Vemo BN, Kenfack A, Ngoula F, Akono Nantia E, Njieudeu Ngaleu CC, Nounamo Guiekep AJ, Megnimeza Tsambou AM, Nana Yidjeu F, Patrick Nelo C, Teguia A. Toxicity and reproductive parameter impairment of cypermethrin in male guinea pig (Cavia porcellus). Turkish J Agric-Food Sci Technol. 2018;6(2):130-5.

Vemo BN, Kenfack A, Ngoula F, Nantia EA, Kodjio N, Guiekep AJN, Tsambou AMM, Teguia A. Effects of ethanol extract of Bersama engleriana leaves on oxidative stress and reproductive parameters in male Guinea pig (Cavia porcellus) exposed to cypermethrin. Int J Biol Chem Sci. 2017;11(5):2243-53.

Venkatachalam RN, Singh K, Marar T. Phytochemical screening and in vitro antioxidant activity of Psidium guajava. Free Rad Ant. 2012;2(1):31-6.

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