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In Vitro and In Vivo Studies on the Efficacy of Zinc-Oxide and Selenium Nanoparticle in Cryopreserved Goat (Capra hircus) Spermatozoa

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Abstract

Different nanoparticles (NPs) are currently being investigated for their potential role as cryoprotectant during semen cryopreservation in several mammalian species. It may be possible to improve semen quality following cryopreservation by supplementation of NPs in the freezing extenders. The present study was carried out in semen collected from four (4) Assam Hill Goat bucks (10 ejaculates per buck) to investigate the effect of supplementing zinc oxide (ZnO) and selenium (Se) NPs in Tris-citric acid-fructose yolk (TCFY) extender on in vitro sperm quality and in vivo fertility rate after freeze-thawing. The size morphology and zeta potential of ZnO and Se NPs were evaluated prior to its incorporation in the freezing extender. Qualified semen samples (> 70% progressive motility) were divided into five (5) aliquots and then diluted in TCFY extender containing ZnO and Se NP supplementation at different concentrations (T0, control; T1, 0.1 mg/mL ZnO NPs; T2, 0.5 mg/mL ZnO NPs; T3, 0.5 µg/mL Se NPs; and T4, 1 µg/mL Se NPs). Diluted semen was packed in 0.25 mL straws and then stored in liquid nitrogen. After thawing, post-thaw in vitro sperm attributes were evaluated. Finally, the effect of NPs on in vivo fertility rate was checked in heat-synched does (n = 70) by artificial insemination (AI) using straws that showed superior results during the in vitro study. Results showed that ZnO and Se NPs were poly-crystalline in nature with particle size below 100 nm (nm). The evaluated post-thaw sperm in vitro attributes were significantly (p < 0.001) higher in T1 in comparison to T0. The antioxidant enzyme activities were significantly (p < 0.001) higher in T1. Lipid peroxidation (LPO) profile was significantly (p < 0.001) lower in T1. Sperm motility and mitochondrial membrane potential (MMP) had a highly significant (r = 0.580, p < 0.05) association in T1. No significant (p > 0.05) differences in pregnancy rates were recorded after AI in the different treatments. In conclusion, extender supplemented with 0.1 mg/mL ZnO NPs improved post-thaw semen quality of goat spermatozoa consequently by increasing activities of endogenous antioxidant enzymes thereby lowering LPO levels. However, improved in vitro outcomes might not correspond to improved field fertility outcomes.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors are grateful to the sophisticated analytical instrument facility (SAIF) of North Eastern Hill University for instrumental support in nanoparticle characterization. The authors also thankfully acknowledge the Director, Indian Council of Agricultural Research (ICAR), Research Complex for North Eastern Hill Region, for granting the necessary permissions to carry out the research work.

Funding

The work was supported by the Indian Council of Agricultural Research, New Delhi, India, through an in-house project.

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Authors and Affiliations

  1. College of Veterinary Science, Assam Agricultural University, Guwahati, India

    Sayed Nabil Abedin, Anubha Baruah, Arundhati Bora, Devo Jyoti Dutta, Sudip Sinha, Shantanu Tamuly & Arundhati Phookan

  2. Semen Biology Laboratory, Division of Animal and Fisheries Science, Indian Council of Agricultural Research (ICAR) Research Complex for North Eastern Hill Region, Ri-Bhoi, Meghalaya, India

    Kishore Kumar Baruah, Govindasamy Kadirvel, Rahul Katiyar, Gautam Khargharia & Sourabh Deori

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  1. Sayed Nabil Abedin
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Contributions

Anubha Baruah, Kishore Kumar Baruah, and Sourabh Deori conceptualized the experimental design, coordinated the study, discussed the results, and gave important contributions to the writing. Sayed Nabil Abedin performed the in vitro trials in the laboratory, compiled the results, did the statistical analyses, and compiled the draft version of the manuscript. Govindasamy Kadirvel and Rahul Katiyar supervised the work. Arundhati Bora, Devo Jyoti Dutta, Sudip Sinha, Shantanu Tamuly, and Arundhati Phookan formatted and edited the final manuscript version.

Corresponding author

Correspondence to Sourabh Deori.

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Competing interests

The authors declare no competing interests.

Ethical Consent and Animal Use

A written approval of the Animal Ethics Committee of the institute was obtained prior to the onset of the experiment (770/GO/Re/S/03/CPCSEA/FVSc/AAU/IAEC/21–22/938).

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Abedin, S.N., Baruah, A., Baruah, K.K. et al. In Vitro and In Vivo Studies on the Efficacy of Zinc-Oxide and Selenium Nanoparticle in Cryopreserved Goat (Capra hircus) Spermatozoa. Biol Trace Elem Res 201, 4726–4745 (2023). https://doi.org/10.1007/s12011-022-03551-6

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