Globally, countries including China have initiated an extensive gene bank conservation program focused on safeguarding crucial animal genetic resources, with specific emphasis on germplasm cells, notably semen. A fundamental aspect of establishing sperm banks relies on the selection of semen collection methods, and its recognized as an indispensable tool in this process [1]. Therefore, the choice of semen collection methods can significantly impact semen quality across various species [[2], [3], [4], [5], [6], [7], [8], [9]]. In small ruminants, two prevalent semen collection techniques stand out: electroejaculation (EE) and artificial vagina (AV) [10]. Each approach presents distinct advantages and drawbacks. Nevertheless, the AV method is the preferred semen collection technique [11], although prior training are required for male subjects [12]. On the other hand, EE could be a valuable procedure, especially for untrained males and wild animals [[13], [14], [15], [16]]. Nonetheless, it has been observed that administering electrical stimulation exceeding six volts during EE may lead to elevated stress levels in male animals [[17], [18], [19]]. Previous studies have shown that ejaculates obtained via EE exhibit comparable volume, motility, viability, and sperm concentration to those collected using AV [20]. In contrast, EE-collected semen has been reported to exhibit a larger volume of seminal plasma compared to that obtained through AV [1], potentially influencing the response to cryopreservation. Furthermore, León et al. [6] found no significant impact of EE and AV on mass and individual sperm motility in cattle. Numerous investigations have also reported the influence of semen collection on sperm quality and fertility [21,22]. The composition and concentration of proteins in SP may vary depending on the collection methods [23]. Specifically, in various mammalian species, certain SP proteins have been identified for their roles in stabilizing sperm membrane, regulating sperm motility, viability, and capacitation, as well as facilitating sperm fertilization [[24], [25], [26], [27], [28], [29]].

Over the last decade, new technologies, such as proteomics, have been incorporated to study the sperm biology [30]. Moreover, some SP-derived proteins have been utilized as biomarkers for sperm functionality, and even fertility [[31], [32], [33], [34], [35]]. It has also been observed that SP proteins differ based on the applied collection methods [36,37]. For example, SP from rams collected by AV showed a lower concentration of low molecular weight proteins as compared to EE [38]. Additionally, using a Tandem Mass Tag (TMT) peptide labeling combined with a LC–MS/ MS identified, a total of 866 proteins in the SP collected with AV from donkey [39]. In stallions, and by using the high throughput LC/MS-MS approach, a total of 1687 proteins were found in SP from AV collected semen [40]. Furthermore, using 2D-PAGE coupled with mass spectrometry, Rego et al. [37], found 48 proteins in SP collected by EE. Nevertheless, debates persist regarding sperm quality and SP proteomes concerning semen collection methods.

Hence, to the best of our knowledge, this study represents the first attempt using advanced technology to evaluate the differential impact of semen collection methods (EE and AV) on semen quality and SP proteomes in Yunshang black goats, which is a newly recognized breed in China. These findings will hold the potential to offer valuable insights into the underlying mechanisms of each semen collection method, assisting in the selection of appropriate techniques for goat semen preservation and contributing to the conservation of goats' breeds.