The collection of gametes from recently deceased domestic and wildlife mammals has been well documented in the literature. Through the utilization of gametes recovered postmortem, scientists have successfully produced embryos in 10 different wildlife species, while in 2 of those, offspring have also been born. Thus, the collection of gametes from recently deceased animals represents a valuable opportunity to increase genetic resource banks, obviating the requirement for invasive procedures. Despite the development of several protocols for gamete collection, the refinement of these techniques and the establishment of species–specific protocols are still required, taking into account both the limitations and the opportunities. In the case of wildlife, the optimization of such protocols is impeded by the scarcity of available animals, many of which have a high genetic value that must be protected rather than utilized for research purposes. Therefore, optimizing protocols for wildlife species by using domestic species as a model is crucial. In this review, we focused on the current advancements in the collection, preservation, and utilization of gametes, postmortem, in selected species belonging to Equidae, Bovidae, and Felidae, both domestic and wildlife.

Canine and feline epididymal semen provide an additional source of gametes to preserve the genetics of valuable breeding dogs and tomcats, especially for those that fail to ejaculate, need castration as a therapy or die unexpectedly. Moreover, since it is quite common to perform castration of non-breeding dogs and cats, the development of a gene bank of epididymal semen collected after castration would greatly contribute to increase the genetic diversity in dogs and cats. Collection and cryopreservation of epididymal semen necessitates a full understanding of the function of the epididymis and of the characteristics of epididymal spermatozoa as opposed to ejaculated semen. During collection of epididymal semen, specific factors may have a negative effect on epididymal semen quality and freezability. Accordingly, the elimination of these triggers could enhance epididymal semen freezability and consequently positively influence post-thaw semen quality and outcome for different ARTs.

Small animal reproduction (SAR) is a main research field in veterinary medicine and bibliometric analyses are useful to investigate trends in specific research areas. The objective of the present study was to conduct a bibliometric analysis of the literature of the last decade on SAR. A search equation was created, and documents were retrieved from the Web of Science database. Documents were manually revised, categorized and R software version 4.1.2 with Bibliometrix R package version 3.1 and MS Excel were used to perform the analyses. The included documents (n = 1470) were mainly research articles (78%). The top countries for the number of documents and citations were Brazil, United States, Italy, Poland, and Korea. These also account for the most prolific authors and institutions. Analyses by author keywords, categories, and recent reviews of the literature suggest that research on the canine species is more abundant than research on the feline one and that reproductive biotechnologies are a main research focus. Some clinical topics are still considered niche or neglected themes (e.g., semen collection in tomcats, neonatology). However, heterogeneity and ambiguity in keywords and categories are undeniable. This study offers interesting insights, providing definitions for main keywords in the field of SAR.

The iSperm^® is a portable device for semen analysis. This study aimed to investigate its correlation with a conventional computer-assisted sperm analyzer (ISAS^®v1) for the assessment of semen concentration and kinematic parameters in dogs (n = 224). The intra-assay variability of both devices and their ability to estimate semen concentration at a fixed value of 40 × 10⁶/mL were also investigated. Results showed that the intra-assay variability was lower for the ISAS^®v1 for all parameters compared to the iSperm^®. Hence, iSperm^® estimates were more variable in-between fields. Both the iSperm^® and the ISAS^®v1 were not reliable in estimating semen concentration (ISAS^®v1: median 30 × 10⁶/mL, interquartile range (IQR) 12, p < 0.01; iSperm^®: median 35.12 × 10⁶/mL, IQR 11.11, p < 0.01). Finally, positive correlations were found between both devices with stronger correlations obtained when four fields were analyzed by the iSperm^®. However, the low number of spermatozoa analyzed per field and the inability to avoid artifacts are downsides that currently limit the reliability of the iSperm^®. Therefore, the software of iSperm^® needs some improvement to make it a valid and practical alternative to automated computerized systems for the analysis of canine semen.

This study evaluated how semen selection by single layer centrifugation (SLC) with Canicoll affects semen freezability in dogs. A total of eighteen ejaculates, collected from dogs with optimal and suboptimal semen quality (optimal: normal morphology (NM) ≥ 80%, n = 9; suboptimal: NM between 60 and 79%, n = 9), were divided into two aliquots and subjected to standard centrifugation or SLC before cryopreservation. Motility, NM, membrane integrity, mitochondrial membrane potential (MMP), and DNA integrity were improved in fresh samples after SLC, regardless of semen quality, but at the expense of some good quality spermatozoa. After thawing, NM and membrane integrity were improved in SLC-selected semen in both semen qualities. Interestingly, MMP was also higher but only in optimal quality semen. Still, spermatozoa from suboptimal quality semen did not survive freezing to the same extent as spermatozoa from optimal quality semen, even after selecting superior spermatozoa. Semen selection with Canicoll is, therefore, an effective technique to isolate a subpopulation of high-quality spermatozoa and obtain sperm samples of better quality after thawing, but is not sufficient to improve the intrinsic inferior freezability of suboptimal quality semen. So far, eighteen pups were born after insemination with SLC-selected frozen-thawed semen, proving that these selected spermatozoa remain fertile.