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. 2023 Jun 5;13:9102. doi: 10.1038/s41598-023-35541-6

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Sperm Functional Parameters. The buffalo spermatozoa were exposed to 0.5, 1, 2, 5, 10 and 20 μM of Omethoate, Paraoxon methyl, TCPy for the assessment of membrane integrity (A–C), capacitation status (D–F), mitochondrial membrane potential (G–I) and lipid peroxidation (J–L) using fluorescent staining. CFDA with PI (see text) was used to assess the membrane integrity of the spermatozoa that were categorized as live, dead or moribund. CTC (see text) was used to categorize the spermatozoa as non-capacitated (NC), capacitated (C), or acrosome-reacted (AR). JC-1 (see text) was used to calculate the percentage of spermatozoa with high and low mitochondrial membrane potential while BODIPY (see text) was used to distinguish the spermatozoa with high and low lipid peroxidation (LPO).