Comparative effects of melatonin and melatonin nanoparticle for cryopreservation of Barbari Buck semen in the summer season

P  KUMAR; S K  MAURYA; R  KUMAR; R  KUMAR; R  KUMAR; RISHIKANT; R  DIWAKAR; K D  SINGH

doi:10.56093/ijans.v95i9.171426

Authors

P KUMAR College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya-224229, UP, India
S K MAURYA College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya-224229, UP, India
R KUMAR College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya-224229, UP, India
R KUMAR College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya-224229, UP, India
R KUMAR College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya-224229, UP, India
RISHIKANT College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya-224229, UP, India
R DIWAKAR College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya-224229, UP, India
K D SINGH College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya-224229, UP, India

https://doi.org/10.56093/ijans.v95i9.171426

Keywords:

Antioxidants, Buck, Cryopreservation, Fertility Assessment, Melatonin, Nanoparticles, Semen Quality

Abstract

The study aimed to evaluate potential of melatonin and melatonin loaded nanoparticles to mitigate cryopreservation-induced cryo-damage using sperm related parameters of Barbari Buck semen. The pooled ejaculates were diluted using Tris-Egg-Yolk-Citric-Acid Fructose-Glycerol (TEYCAFG) extender and subsequently partitioned into three aliquots i.e. CON (without additives), MT (fortified with 1.0mM melatonin), and MTN (fortified with 5.0µM melatonin nanoparticles. Various seminal attribute viz. motility, viability, HOS reactive sperm, and the levels of GOT, and GPT was evaluated at pre-freeze and post-thaw stage. Significantly (P<0.05) higher motility and liveabilty in MTN (58.56±2.17 and 53.80±8.73%) and MT (55.95±2.6 and 51.84±8.72%) groups were recorded as compared those of CON (46.14±0.25 and 45.56±7.49%). Similarly, significantly (P<0.05) higher HOS reactive sperms and sperm acrosomal integrity was noted in MTN (57.99±2.11 and 67.24±1.22%) as compared to MT (54.18±2.12 and 64.92±2.05%) and CON (50.14±0.41and 61.28±0.26%). In addition, abnormality, and enzyme leakage (GPT and GOT) was significantly (P<0.05) declined in MTN (6.29±0.48%, 25.46±0.74 and 127.30±1.86 µM/L) as compared to MT (7.14±0.72%, 27.94±1.04 and 149.30±2.11 µM/L) and CON 8.50±0.11%, 30.14±0.00 and 157.70±0.00 µM/L). Overall, the incorporation of 5 μM melatonin nanoparticles in the semen dilutor significantly improved post-thaw seminal attributes of Barbari buck semen.

Downloads

Download data is not yet available.

References

Abdolvahab MH, Karimi P, Mohajeri N, Abedini M and Zare H. 2024. Targeted drug delivery using nanobodies to deliver effective molecules to breast cancer cells: the most attractive application of nanobodies. Cancer Cell International 24(1): 67.

Akhtarshenas B, Kowsar R, Hajian M, Vash NT, Soltani L, Mahdavi AH and Esfahani MHN. 2024. ρ-Coumaric acid-zinc oxide nanoparticles improve post-thaw quality of goat spermatozoa and developmental competence of fertilized oocytes in vitro. Scientific Reports 14(1): 31971.

Alam K, Srivastava S, Singh B, Kumar R, Kumar R and Sakhahari D S. 2024a. Cryopreservation of bovine semen using extract of Cinnamomum zeylanicum. Cryoletters 45(3): 168–76.

Alam K, Srivastava S, Verma S K, Verma A K, Saurabh, Kumar R, Kumar R, Jaiswal, A, Alam A and Rana T. 2024b. The effect of different concentrations of melatonin on the quality of frozen and thawed semen of Jamnapari goat. Reproduction in Domestic Animals 59(11): e14743.

Aranarochana A, Sirichoat A, Pannangrong W, Wigmore P and Welbat J U. 2021. Melatonin Ameliorates Valproic Acid-Induced Neurogenesis Impairment: The Role of Oxidative Stress in Adult Rats. Oxidative Medicine and Cellular Longevity 2021(1): 9997582.

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): 1220–33.

Cao B, Qin J, Pan B, Qazi I H, Ye J, Fang Y and Zhou G. 2022. Oxidative stress and oocyte cryopreservation: recent advances in mitigation strategies involving antioxidants. Cells 11 (22): 3573.

Casao A, Pena-Delgado V and Pérez-Pe R. 2025. From spermatogenesis to fertilisation: The role of melatonin on ram spermatozoa. Domestic Animal Endocrinology 91: 106916.

Chen R, Sun G, Xu L, Zhang X, Zeng W and Sun X. 2022. Didymin attenuates doxorubicin-induced cardiotoxicity by inhibiting oxidative stress. Chinese Herbal Medicines 14(1): 70–78.

Danso F, Iddrisu L, Lungu S E, Zhou G and Ju X. 2024. Effects of heat stress on goat production and mitigating strategies: a review. Animals 14(12): 1793.

Deka B C and Rao A R. 1986. Motility of buck spermatozoa during preservation at 5° C with and without seminal plasma. The Indian Veterinary Journal 63:169–70.

Evans G and Maxwell W C. 1987. Salamons' artificial insemination of sheep and goats (No. Ed. 2, pp. xi+-194).

Fayez E, Samir H, Youssef F S, Salama A and El Sayed M A. 2024. Administration of melatonin nanoparticles improves testicular blood flow, echotexture of testicular parenchyma, scrotal circumference, and levels of estradiol and nitric oxide in prepubertal ossimi rams under summer heat stress. Veterinary Research Communications 48(6): 3953–65.

Franco P I R, do Carmo Neto J R, Braga Y L L, de Lima Pedroso B, Martins J A, Rocha V L, Amaral A C, Martins D B, Ruiz K C, Pereira J X and e Silva J R M. 2025. Melatonin-loaded lecithin and chitosan nanoparticles are cytotoxic to 4 T1 breast cancer cells and safe in a BALB/c mouse model. International Journal of Biological Macromolecules 311(1):143401.

Galano A, Tan D X and Reiter R J. 2018. Melatonin: a versatile protector against oxidative DNA damage. Molecules 23(3): 530.

Galián S, Peinado B, Almela L, Poto Á and Ruiz S. 2023. Post-thaw quality of spermatozoa frozen with three different extenders in the Murciano Granadina goat breed. Animals 13(2): 309.

Gangwar C, Kharche S D, Ranjan R, Kumar S, Goel A K, Jindal S K and Agarwal S K. 2015. Effect of vitamin C supplementation on freezability of Barbari buck semen. Small Ruminant Research 129: 104–7.

Ghosh V, Mukherjee A and Chandrasekaran N. 2015. Optimization of process parameters to develop nanoemulsion by ultrasound cavitation. Journal of Pure and Applied 53.

Hafez E S E and Hafez B. 2000. Transport and Survival of Gametes In Reproduction in Farm Animal. 7 th. Ed. ESE Hafez, B. Hafez. Kiawah Island, South Carolina, USA.

Hai E, Li B, Zhang J and Zhang J. 2024. Sperm freezing damage: the role of regulated cell death. Cell Death Discovery 10(1): 239.

Jeyendran R S, Van der Ven H H, Perez-Pelaez M, Crabo B G and Zaneveld L J D. 1984. Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. Reproduction 70(1): 219–28.

Jomova K, Alomar S Y, Alwasel S H, Nepovimova E, Kuca K and Valko M. 2024. Several lines of antioxidant defense against oxidative stress: antioxidant enzymes, nanomaterials with multiple enzyme-mimicking activities, and low-molecular-weight antioxidants. Archives of Toxicology 98(5): 1323–67.

Joseph T T, Schuch V, Hossack D J, Chakraborty R and Johnson E L. 2024. Melatonin: the placental antioxidant and anti-inflammatory. Frontiers in Immunology 15: 1339304.

Kumar P, Upadyay R C, Kumar R, Chaudhary P K, Maurya P K, Kumar R, Yadav V and Kumar M. 2018. Evaluation and comparison of heat stress indices for cattle and buffaloes. International Journal of Current Microbiology and Applied Sciences 7: 292–98.

Li T, Zheng, Y, Li T, Guo M, Wu X, Liu R, Liu Q, You X, Zeng W and Lv Y, 2022. Potential dual protective effects of melatonin on spermatogonia against hexavalent chromium. Reproductive Toxicology 111: 92–105.

Liu Y, Liang Y, Yuhong J, Xin P, Han J L, Du Y, Yu X, Zhu R, Zhang M, Chen W and MaY. 2024.Advances in Nanotechnology for Enhancing the Solubility and Bioavailability of Poorly Soluble Drugs. Drug Design, Development and Therapy 18: 1469–95.

Lu X, Fei L, Li Y, Du J, Ma W, Huang H and Wang J. 2023. Effect of different plant growth regulators on callus and adventitious shoots induction, polysaccharides accumulation and antioxidant activity of Rhodiola dumulosa. Chinese Herbal Medicines 15(2): 271–77.

Makris A, Alevra, A I, Exadactylos A and Papadopoulos S. 2023. The Role of Melatonin to Ameliorate Oxidative Stress in Sperm Cells. International Journal of Molecular Sciences 24(20): 15056.

Milewska A J, Kuczyńska A, Pawłowski M, Martynowicz I, Deluga-Białowarczuk S, Sieczyński P, Kuczyński W and Milewski R. 2024. The Influence of Cryopreservation on Sperm Morphology and Its Implications in Terms of Fractions of Higher-Quality Sperm. Journal of Clinical Medicine 13(24): 7562.

Monteiro K K A C, Shiroma M E, Damous L L, Simões M D J, Simões R D S, Cipolla-Neto J, Baracat E C and Soares-Jr J M. 2024. Antioxidant actions of melatonin: a systematic review of animal studies. Antioxidants 13(4): 439.

Paulenz H, Soderquist L, Pérez-Pé R and Berg K A. 2002. Effect of different extenders and storage temperatures on sperm viability of liquid ram semen. Theriogenology 57(2): 823–36.

Peng L, Ru M and Liang Z. 2022. Variation of photosynthesis, secondary metabolites and antioxidant activities in third generation of spaceflight-induced Salvia miltiorrhiza. Chinese Herbal Medicines 14(4): 592–601.

Peris-Frau P, Soler A J, Iniesta-Cuerda M, Martín-Maestro A, Sánchez-Ajofrín I, Medina-Chávez D A, Fernández-Santos M R, García-Álvarez O, Maroto-Morales A, Montoro V and Garde J J. 2020. Sperm cryodamage in ruminants: understanding the molecular changes induced by the cryopreservation process to optimize sperm quality. International Journal of Molecular Sciences 21(8): 2781.

Sari R P, Nugraha C D and Suyadi S. 2024. Pengaruh thawing terhadapkualitas semen kambingboermenggunakanpengencer tris-aminomethane yang disuplementasiantioksidan quercetin. Jurnal Sain Peternakan Indonesia 19(1): 11-20.

Sarıkaya E and Dogan S. 2020. Glutathione Peroxidase in Health. Glutathione System and Oxidative Stress in Health and Disease 49.

Sikka S C, Rajasekaran M A and Hellstrom W J. 1995. Role of oxidative stress and antioxidants in male infertility. Journal of Andrology16: 464–8.

Sikka S C. 2001. Relative impact of oxidative stress on male reproductive function. Current Medicinal Chemistry 8(7): 851–62.

Sun W, Jiang S, Su J, Zhang J, Bao X, Ding R, Shi P, Li S, Wu C, Zhao G and Cao G. 2021. The effects of cryopreservation on the acrosome structure, enzyme activity, motility, and fertility of bovine, ovine, and goat sperm. Animal Reproduction 17(4): e20200219.

Swift M L. 1997. GraphPad prism, data analysis, and scientific graphing. Journal of Chemical Information and Computer Sciences 37(2): 411–12.

Tan D X, Manchester L C, Esteban-Zubero E, Zhou Z and Reiter R J. 2015. Melatonin as a potent and inducible endogenous antioxidant: synthesis and metabolism. Molecules 20(10): 18886–906.

Tanga B M, Qamar A Y, Raza S, Bang S, Fang X, Yoon K and Cho J. 2021. Semen evaluation: Methodological advancements in sperm quality-specific fertility assessment- A review. Animal Bioscience 34(8): 1253.

Tucker C B, Rogers A R and Schütz K E. 2008. Effect of solar radiation on dairy cattle behaviour, use of shade and body temperature in a pasture-based system. Applied Animal Behaviour Science 109(2–4): 141–54.

Upadhyay R C, Sirohi S, Ashutosh, Singh S V, Kumar A and Gupta S K. 2009. In: Global Climate Change and IndianAgriculture (Edited by PK Aggarwal). Published by ICAR, New Delhi. Pp. 104–7.

Wang H, Tian Y, Tan H, Zhou M, Li M, Zhi Y, Shi Y and Li X. 2023. Research and application of leek roots in medicinal field. Chinese Herbal Medicines 15(3): 391–97.

Wang Y, Fu X and Li H. 2025. Mechanisms of oxidative stress-induced sperm dysfunction. Frontiers in Endocrinology 16: 1520835.

West J W, Mullinix B G and Bernard J K. 2003. Effects of hot, humid weather on milk temperature, dry matter intake, and milk yield of lactating dairy cows. Journal of Dairy Science 86(1): 232–42.

Xi H, Gao X, Qiu L, Wang Y, Qiu Y, Tao Z, Hu M, Jiang X, Yao Q, Kou L, Zhao J and Chen R. 2025. Melatonin-loaded nanoparticles protecting human sperm from oxidative stress during cryopreservation. Expert Opinion on Drug Delivery 22(7): 1043–55.

Xu Y, Sun S, Wang M, Shen W, Wang L, Ren C, Ling Y, Zhang Z and Cao H. 2025. Dexamethasone and azithromycin enhance goat sperm preservation quality by regulating lipid metabolism. Theriogenology 231:197–209.

Xue S H, Xu B B, Yan X C, Zhang J X and Su R. 2025. Sperm Membrane Stability: In-Depth Analysis from Structural Basis to Functional Regulation. Veterinary Sciences 12(7): 658.

Yang B, Xia L, Deng R, Wu L, Zhang Z, Wu X, Ding T, Zhao Y, Huang J and Huang Z. 2025. Impact of sperm DNA fragmentation index on assisted reproductive outcomes: a retrospective analysis. Frontiers in Endocrinology 15:1530972.

Zhou D, Chang W, Qi J, Chen G and Li N. 2023. Lung protective effects of dietary malate esters derivatives from Bletilla striata against SiO2 nanoparticles through activation of Nrf2 pathway. Chinese Herbal Medicines 15(1): 76–85.

Zhu L, Zhang Q, Hua C and Ci X. 2023. Melatonin alleviates particulate matter-induced liver fibrosis by inhibiting ROS-mediated mitophagy and inflammation via Nrf2 activation. Ecotoxicology and Environmental Safety 268: 115717.

Zou J, Wei L, Li D, Zhang Y, Wang G, Zhang L, Cao P and Li G. 2022. Study on cryopreservation of Guanzhong dairy goat semen with bovine semen seminal plasma. Theriogenology 189: 113–17.