Reproductive Biotechnologies Research Articles

The Sixth Mass Extinction and Amphibian Species Sustainability Through Reproduction and Advanced Biotechnologies, Biobanking of Germplasm and Somatic Cells, and Conservation Breeding Programs (RBCs)

Primary themes in intergenerational justice are a healthy environment, the perpetuation of Earth’s biodiversity, and the sustainable management of the biosphere. However, the current rate of species declines globally, ecosystem collapses driven by accelerating and catastrophic global heating, and a plethora of other threats preclude the ability of habitat protection alone to prevent a cascade of amphibian and other species mass extinctions. Reproduction and advanced biotechnologies, biobanking of germplasm and somatic cells, and conservation breeding programs (RBCs) offer a transformative change in biodiversity management. This change can economically and reliably perpetuate species irrespective of environmental targets and extend to satisfy humanity’s future needs as the biosphere expands into space. Currently applied RBCs include the hormonal stimulation of reproduction, the collection and refrigerated storage of sperm and oocytes, sperm cryopreservation, in vitro fertilization, and biobanking of germplasm and somatic cells. The benefits of advanced biotechnologies in development, such as assisted evolution and cloning for species adaptation or restoration, have yet to be fully realized. We broaden our discussion to include genetic management, political and cultural engagement, and future applications, including the extension of the biosphere through humanity’s interplanetary and interstellar colonization. The development and application of RBCs raise intriguing ethical, theological, and philosophical issues. We address these themes with amphibian models to introduce the Multidisciplinary Digital Publishing Institute Special Issue, The Sixth Mass Extinction and Species Sustainability through Reproduction Biotechnologies, Biobanking, and Conservation Breeding Programs.

Reproductive Biotechnology Tools as a Game Changer for the Dairy Sector: The Case of Sexed Semen and Estrus Synchronization to Produce Seedstock Dairy Heifers

Human curiosity about the sexual orientation of living things has a long history. For thousands of years, livestock owners wanted to find a suitable way to predict the sex of the progeny to be born for their herds on their farm. Thanks to the development of semen sexing technology, animal breeding has undergone a revolution, which allows farmers to control the sex of their offspring. Sexed semen enhances overall productivity and also assists farmers in meeting the rising demand for high-quality dairy animals by enabling them to selectively generate seedstock female calves. The technology is a one-stop solution to enhance breed improvement especially in a country like Ethiopia where 97 percent of the cattle population is indigenous, which is not selected for milk production and results in undernourishment for animal source origin diet, including milk and meat. The benefits of using sexed semen include sex selection, improved genetic selection, enhanced breeding effectiveness, better control over herd dynamics, greater financial gains, environmental sustainability, and improved animal welfare. Hence, with the finding of this study, an overall result of a 98% response rate 95% female skewness, and 79% conception rate was recorded after being inseminated with sexed semen. The finding of the response rate is closer to the results reported by another researcher, while the average conception rate is comparatively higher. This could be due to careful animal selection, the use of fertile quality semen, proper heat detection, and inseminating at an optimum window of time. In general, from the piece of this study, it is possible to foresee and conclude that the adoption and application of sexed semen technology is a groundbreaking and game-changer technology for the dairy industry in a country like Ethiopia, which overcomes the limitations of improved crossbred dairy cattle, replacement heifers, and high milk prices on the one hand, a shortage of grazing land, ...

Dystocia in Dairy Cows and Heifers: A Review with a Focus on Future Perspectives

Dairy cow management has evolved tremendously in recent decades, particularly regarding reproductive techniques. The widespread adoption of synchronization protocols, sexed semen, beef semen in dairy cows, reproductive biotechnologies such as in vivo-derived (IVD) or in vitro-produced embryos (IVP), and precision livestock farming is transforming the daily practices of dairy farmers and veterinarians. These implementations are typically adapted in different ways when applied to the breeding of heifers or cows. Considering these developments, dystocia—a significant welfare and productivity concern—may warrant reevaluation. The aforementioned changes are likely to have a substantial impact on its prevalence, severity, and outcome. This review aims to address the main aspects of dystocia in dairy cows and heifers, with a particular focus on the potential impact of recent advances in reproductive and calving management.

Opportunities for CRISPR-Cas9 application in farm animal genetic improvement.

CRISPR-Cas9 has emerged as a powerful tool in livestock breeding, enabling precise genetic modifications to address genetic diseases, enhance productivity, and develop disease-resistant animal breeds. A thorough analysis of previous research highlights the potential of CRISPR-Cas9 in overcoming genetic disorders by targeting specific mutations in genes. Furthermore, its integration with reproductive biotechnologies and genomic selection facilitates the production of gene-edited animals with high genomic value, contributing to genetic enhancement and improved productivity. Additionally, CRISPR-Cas9 opens new avenues for developing disease-resistant livestock and creating innovative breeding models for high-quality production. A key trend in the field is the development of multi-sgRNA vectors to correct mutations in various genes linked to productivity traits or certain diseases within individual genomes, thereby increasing resistance in animals. However, despite the potential advantages of CRISPR-Cas9, public acceptance of genetically modified agricultural products remains uncertain. Would consumers be willing to purchase such products? It is essential to advocate for bold and innovative research into genetically edited animals, with a focus on safety, careful promotion, and strict regulatory oversight to align with long-term goals and public acceptance. Continued advancements in this technology and its underlying mechanisms promise to improve poultry products and genetically modified livestock. Overall, CRISPR-Cas9 technology offers a promising pathway for advancing livestock breeding practices, with opportunities for genetic improvement, enhanced disease resistance, and greater productivity.

DYSTOCIAS IN CATTLE CAUSES AND CONSEQUENCES: A brief integrative literature review

Among domestic animals, cattle are the species that present problems with dystocia, bringing great economic losses to the producer. Dystocia can occur for several reasons, from a slight delay in the normal onset of labor or even the impossibility of giving birth, almost always requiring the intervention of a veterinarian, and can only be corrected with obstetric maneuvers, in the most severe cases, cesarean section or when the product is dead, fetotomy. The study aims to examine research published between 2017 and 2023, preferably, that discusses challenges in childbirth, reproductive biotechnologies, reproductive diseases, livestock management and the effect of dystocia on animal production and health. This study is characterized by being a literature review whose general objective is to discuss the importance of knowledge of dystocia in childbirth is fundamental to carry out the appropriate intervention. After the process of choosing and defining the theme, a bibliographic search was carried out in the Google Scholar database. The search terms were: difficult calving, reproductive biotechnologies, twin birth, embryo transfer, reproductive pathologies, fetal development, reproductive physiology, cattle management. From the literature review, it was possible to note the agreement between the literature and the practices carried out in the field by veterinarians, with regard to placenta retention and uterine torsion. The occurrence of placenta retention is considered a very common episode and many factors contribute to the development of this disease, which causes many secondary problems in the cattle herd.

Success of Artificial Insemination in Rural Area Cows

Biotechnologies of reproduction like artificial insemination were identified as a mean to intensify the local production. Therefore, some factors are still responsible to the failure of their applications in rural area. This survey was conducted from April to June 2023, to evaluate the effects of age, body weight, body condition score (BCS), calving rank, postpartum to artificial insemination interval and feed complementation on the success rate of artificial insemination in females Goudali. Thus, activities took place at two peri-urban livestock farms in the town of Ngaoundéré, in the Adamaoua region of Cameroon where 34 cows were concerned. The selected cows have an average of 8.21±2.41 years old, a body weight of 343±52.28 kg, a body condition score of 3.23±0.55, a calving rank of 2.21±1.22 and a postpartum to insemination interval of 4.39±1.40 months. The induction and synchronization of estrus were carried out based on the PRID-PGF2α-PMSG protocol which allowed to obtain a synchronization rate of 97.06% and the insemination were done 56-57 hours after the previous operation, with an imported semen of Brahman bull. During rectal palpation and early pregnancy diagnosis based on progesterone level measurement, the result shown that, 20 cows out of 33 inseminated were pregnant. The success rate of artificial insemination recorded was 60.6%. Amongst all the studied factors, analysis shown no significant (P> 0.05) effect on the success rate after artificial insemination. Thus, recommendations were made to popularize the use of artificial insemination and early pregnancy diagnosis test as a mean to improve reproductive performances of local farms. However, studies must be done to evaluate the cost of their applications in rural areas.

Methodological approaches in vitrification: Enhancing viability of bovine oocytes and invitro-produced embryos.

Cryopreservation of bovine oocytes and embryos is essential for long-term preservation and widespread distribution of genetic material, particularly in bovine invitro embryo production, which has witnessed substantial growth in the past decade due to advancements in reproductive biotechnologies. Among current cryopreservation methods, vitrification has emerged as the preferred cryopreservation technique over slow freezing for preserving oocytes and invitro-produced (IVP) embryos, as it effectively addresses membrane chilling injury and ice crystal formation. Nonetheless, challenges remain and a simple and robust vitrification protocol that guarantees the efficiency and viability after warming has not yet been developed. Furthermore, although slow cooling can easily be adapted for direct transfer, an easier and more practical vitrification protocol for IVP embryos is required to allow the transfer of IVP embryos on farms using in-straw dilution. In addition, the susceptibility of bovine oocytes and embryos to cryoinjuries highlights the need for novel strategies to improve their cryotolerance. This manuscript examines various methodological approaches for increasing the viability of bovine oocytes and IVP embryos during vitrification. Strategies such as modifying lipid content or mitigating oxidative damage have shown promise in improving cryotolerance. Additionally, mathematical modelling of oocyte and embryo membrane permeability has facilitated the rational design of cryopreservation protocols, optimizing the exposure time and concentration of cryoprotectants to reduce cytotoxicity.

Sperm Metabolism.

Bioenergetics plays a crucial role in sperm functions, including motility, capacitation-related protein modifications, oocyte recognition and interaction, all of which are essential for fertilization. Sperm metabolism is recognized as flexible, responding to environmental cues and energetic demands during ejaculation, the journey along the female tract, and until fertilization. Recent studies suggest that sperm metabolic functions are relevant beyond fertilization and may influence zygote and embryo development, impacting paternal-derived effects on offspring development and health. In recent years, sperm metabolic functions and homeostasis have gained increasing interest in male reproduction research. Given the crucial implications of sperm metabolism on fertility-related processes, this field is of interest not only in human male fertility but also in livestock research, semen conservation, and assisted reproductive techniques. Newly developed assessment tools are allowing a better understanding of sperm metabolism under different conditions and identifying species-specific peculiarities. This review aims to discuss the current knowledge of mammalian sperm metabolism, focusing on species-specific features, changes during the sperm journey, and potential contributions to translational research and reproductive biotechnologies. Furthermore, we propose future perspectives on sperm bioenergetics research.

Why choose the rabbit to work in reproductive technology?

Rabbits have played a significant role in both livestock production and the advancement of reproductive scientific research. Their unique biological traits, including induced ovulation and a reproductive process that closely mirrors that of humans, have been pivotal in their use as a model. Moreover, their body size is perfectly aligned with the 3Rs principles: Replacement, Reduction, and Refinement. Consequently, techniques for gamete collection and embryo recovery, followed by their use in artificial insemination or embryo transfer, are characterized by being minimally invasive. However, refining invitro fertilization and embryo culture techniques continues to present challenges. The incorporation of cutting-edge genomic editing tools, such as CRISPR/Cas9, has reestablished rabbits as essential models in genetic and biomedical research, driving scientific progress. This review aims to describe the most effective reproductive biotechnologies for both male and female rabbits and how these methodologies are in line with the 3Rs principles-Replacement, Reduction, and Refinement-highlighting their significance in conducting ethical research.

In Vitro Embryo Production: Overview

Reproductive biotechnologies, also known as assisted reproductive technologies (ART), have undergone significant development over time, reaching a remarkable level of evolution in the 20th century. Currently, several biotechnologies stand out, among which Artificial Insemination (AI), widely spread globally, Embryo Production (EP) both in vivo and in vitro for embryo transfer, Intracytoplasmic Sperm Injection (ICSI), sexing, among other innovations. In Vitro Production (IVP) of embryos has emerged as the biotechnology that has experienced the greatest development and evolution in recent decades, covering various fields such as research in assisted human reproduction, obtaining transgenic animals for biomedicine, preservation of endangered species and conservation of animals of high genetic value. This research aims to analyze the relevance of IVP, as well as the fundamental elements that make up this complex process. IVP of embryos involves several stages, from oocyte procurement, oocyte selection and in vitro maturation, sperm preparation, in vitro fertilization, to the culture of the resulting zygotes to blastocysts. These blastocysts, once transferable to recipients, culminate in the birth of a new being. However, this process has both advantages and disadvantages, which motivates continuous innovation in the field.

Effect of different concentrations of inulin on ram sperm quality during cryopreservation

BACKGROUND: In reproductive biotechnology, sperm cryopreservation has a vital role to play. Cryopreservation of sperm produces reactive oxygen species (ROS), which disrupt sperm function and structural competence. Numerous protective chemicals, including fructans, have been used during sperm cryopreservation. OBJECTIVES: To evaluate the effect of different concentrations of the fructosan inulin on ram sperm quality parameters, superoxide dismutase (SOD) activity, and malondialdehyde (MDA) production after freezing and thawing. MATERIALS AND METHODS: The pooled samples from four healthy rams were divided into seven equal aliquots and diluted in a Tris-base extender supplemented with 1, 2, 4, 8, 16, and 28 mM of inulin or without inulin supplementation (control). By using liquid nitrogen vapor, the semen was frozen and stored at 196°C. RESULTS: The total motility, viability, and DNA integrity were significantly improved after freezet-hawing with 28 mM inulin, compared to other treatment groups (P<0.05). A Tris-based extender containing 16 and 28 mM of inulin displayed the highest levels of ram sperm membrane integrity when compared with the control (p<0.05). The abnormality of ram sperm was increased during freeze-thawing at control and 1 mM of inulin, compared to 16 and 28 mM of inulin (P<0.05). Additionally, 28 mM of inulin decreased MDA and increased SOD activity in ram sperm in comparison with the other treatments (P<0.05). CONCLUSION: As a result, 28 mM of inulin could be beneficial for the cryopreservation industry and reduce the harmful effects of freeze-thawing on ram sperm.

Multivariate analysis of herd structure and genetic resource indicators in seedstock beef cattle herds.

Genetic, environmental, technological and financial resources are used differently in cattle herds that participate in the same breeding programme. The percentages of calves sired by sires within their own herd or from external herds vary across herds, as do the intensities of use of reproductive biotechnologies. These divergences may be related to differences in the indicators of genetic performance for economic traits. The aim of this study was to determine the factors related to herd structure and genetic resource utilization that exert the greatest influence on the genetic merit of seedstock herds within a Nellore breeding programme. The database comprised 21 factors, along with genomic-enhanced expected progeny differences (GE-EPDs) for growth, reproductive and carcass traits, as well as a selection index of animals from 128 herds. By combining principal component analysis and cluster analysis, we were able to group the herds. We identified statistically significant differences (p < 0.05) in the mean values of the factors, GE-EPDs and genetic trends among the groups of herds. Differences in the percentage of sires from external herds and in sire age between the groups of herds were the factors most associated with differences in mean GE-EPDs and genetic trends. Using young sires from other herds or lineages is an effective strategy in animal breeding. By enhancing genetic variability, this approach does not only improve the genetic quality of herds but also accelerates genetic progress in desired traits over time. Therefore, to ensure the success of this strategy, it is crucial that seedstock herds undergo a thorough selection process aimed at maximizing the genetic potential of future generations of beef cattle.

ACADEMICIAN V. P. BURKAT (1939–2009) – FOUNDER OF MODERN THEORY AND METHODOLOGY OF BREEDING, ORGANIZER OF RESEARCH WORK IN ANIMAL HUSBANDRY IN UKRAINE (to the 85th anniversary of the birthday)

The purpose of the article is to reconstruct the major life milestones and creative path of the recognized scientist in the field of animal husbandry, Doctor of Agricultural Sciences, Professor, Academician of UAAS V. P. Burkat. Research methods are general scientific (analysis, synthesis, classification), special historical (problematic-chronological, comparative-historical), biographical and source studies. The contribution of the scientist to the development of theoretical and methodological principles of breeding, selection, and biotechnology of reproduction of farm animals is substantiated. The important role of V. P. Burkat in the organization of domestic research work in animal husbandry, in particular, in the formation of the Institute of Animal Breeding and Genetics named after M.V. Zubets NAAS was proven. The scientific heritage of the scientist is systematized, the priority of his research on the formation of a systematic concept of the breed, linear breeding, development of genetic and biotechnological bases of selection in animal husbandry, the actualization of the problem of conservation and rational use of the farm animals, etc. is substantiated. The contribution of V. P. Burkat to the formation of the modern theory and methodology of breeding in dairy and beef cattle breeding, the breeding of specialized breeds and types of cattle is revealed. It is shown that among his scientist's methodological developments, the most significant is the method of creating the Ukrainian Red-and-White Dairy breed, which is the basis for the development of other specialized domestic breeds of dairy and beef production directions. The program of activities and achievements of established by V.P. Burkat of the scientific school "Breeding and Biotechnology in Animal Husbandry" is summarized. The scholar's scientific search for the popularization and representation of the history of agricultural science in Ukraine is highlighted.

Reproductive Biotechnology Tools as a Game Changer for the Dairy Sector: The Case of Sexed Semen and Estrus Synchronization to Produce Seedstock Dairy Heifers

Human curiosity about the sexual orientation of living things has a long history. For thousands of years, livestock owners wanted to find a suitable way to predict the sex of the progeny to be born for their herds on their farm. Thanks to the development of semen sexing technology, animal breeding has undergone a revolution, which allows farmers to control the sex of their offspring. Sexed semen enhances overall productivity and also assists farmers in meeting the rising demand for high-quality dairy animals by enabling them to selectively generate seedstock female calves. The technology is a one-stop solution to enhance breed improvement especially in a country like Ethiopia where 97 percent of the cattle population is indigenous, which is not selected for milk production and results in undernourishment for animal source origin diet, including milk and meat. The benefits of using sexed semen include sex selection, improved genetic selection, enhanced breeding effectiveness, better control over herd dynamics, greater financial gains, environmental sustainability, and improved animal welfare. Hence, with the finding of this study, an overall result of a 98% response rate 95% female skewness, and 79% conception rate was recorded after being inseminated with sexed semen. The finding of the response rate is closer to the results reported by another researcher, while the average conception rate is comparatively higher. This could be due to careful animal selection, the use of fertile quality semen, proper heat detection, and inseminating at an optimum window of time. In general, from the piece of this study, it is possible to foresee and conclude that the adoption and application of sexed semen technology is a groundbreaking and game-changer technology for the dairy industry in a country like Ethiopia, which overcomes the limitations of improved crossbred dairy cattle, replacement heifers, and high milk prices on the one hand, a shortage of grazing land, and environmental destruction due to a decreasing number of unwanted cattle population on the other side.

Fixed-Time Artificial Insemination Versus Artificial Insemination at Stalk Detected in Cows And Heifers

Objective: The objective of the present study was to evaluate the pregnancy rate of IACD and IATF in cows and heifers. Theoretical framework: Artificial insemination at a fixed time (IATF) and artificial insemination at detected heat (IACD) are essential tools in livestock farms, improving the production and reproduction of the herd in a short time, considering that differences sometimes occur between the pregnancy rate of cows and heifers with different treatments influencing the production of the herds. Methodology: 194 heifers and 200 Bos taurus x Bos indicus crossbred cows were used. The animals that entered IATF on a random day of the estrous cycle were synchronized with an intravaginal device of Progesterone and Estradiol. The animals that entered IACD were monitored. visual of the characteristic signs of heat twice a day for a period of two hours each, to carry out artificial insemination (AI), the statistical analysis was carried out with the help of the SAS software. Results and discussion: The pregnancy rate among the 215 heifers and cows in the IACD group was 64.00% and 44.00%, respectively, unlike the IATF group with a total of 179 animals, a pregnancy rate of 91.14% for heifers and 78.00% in cows, showing statistically significant differences in follicular size for cows and heifers under treatment with IACD and similar for IATF. Implications of the research: The cyclicity of heifers was efficiently induced in the hormonal protocols with IATF, this group of animals had a higher pregnancy rate and greater follicular diameter unlike the IACD group. Originality/Value: This study contributes to the understanding and effectiveness of using reproductive biotechnologies in livestock herds to increase production and profitability, thus being more sustainable.

Unlocking the potential of propolis: Concentration-dependent effects on Ossimi Ram (Ovis aries) semen quality

Reproductive biotechnology requires successful and efficient cryopreservation of semen since it is a crucial technique for artificial insemination, which preserves and multiplies the superior genetic origins of animals. Propolis, a natural antioxidant produced by honeybees, was examined in this study. Tris-citric-glucose-soybean lecithin extender supplemented with propolis ethanolic extract (P, μg/mL) at 0 (Control), 25 (T1), 50 (T2), 100 (T3), and 200 (T4) was used. The extended semen, prepared in a 1:20 ratio (semen: extender) at 37 °C, was equilibrated for 2 hours at 5 °C and packaged in straws (0.25 ml) before storage in liquid nitrogen at -196 °C. Visual characteristics (e.g., motility, livability, abnormality), DNA integrity, lipid peroxidation, and total antioxidant capacity of semen after thawing with P extract were thoroughly investigated. The results revealed that the low concentration (T1, 25 and T2, 50 µg/ml P) decreased lipid peroxidation, apoptotic sperm, improved sperm characteristics, and maintained the highest sperm cell viability. This resulted in improved reproductive performance and DNA integrity during cryopreservation of Ossimi ram semen. However, the high concentration (T3, 100 and T4, 200 µg/ml P) had the opposite effect and showed a low impact on maintaining cryopreserved Ossimi ram semen. In conclusion, Propolis shows potential for enhancing cryopreservation efficiency, but an optimal dosage is crucial to avoid adverse effects.

The Role of Skin-Derived Somatic Cell and Tissue Cryobanks in the Conservation of Aquatic Mammals.

Anthropogenic actions, especially inadequate waste disposal, cause permanent effects on aquatic fauna, resulting in a significant loss in their population. In this scenario, in situ and ex situ conservation strategies have been developed for these species. Among these strategies is the formation of somatic cell and tissue banks derived from skin collection that act complementarily to other biotechnologies. These banks contain all the information for genomic, genetic, and proteomic analyses. They are useful in the assessment of the toxicity of pollutants on the physiology of the species and regenerative and reproductive biotechnologies. The formation of these cryobanks involves different steps, including cryopreservation, with the optimization of all steps occurring in a species-specific manner. There is a diversity of studies on aquatic mammals; however, a low quantity compared to the number of studies on land mammals, with more than 80% of species still unexplored. This is mainly due to the difficulty of execution and asepsis in collecting skin from aquatic mammals and the in vitro culture, which seems to require more particularities for it to be successful. Therefore, this review aims to address the current scenario and the steps involved in the conservation of somatic cells and tissues derived from aquatic mammal skin, as well as results that have been achieved in recent years and the prospects.

Is it safe to use ovarian follicular fluid from cows seropositive for brucellosis in the production of invitro embryos?

Animal reproduction biotechniques are important tools for the technological advancement of livestock, as they allow the selection of the reproductive potential of superior quality females and males; however, infectious diseases that have a predilection for the reproductive system can be a hindrance for the use of these technologies. Therefore, the present study aimed to detect Brucella spp. in the ovarian follicular fluid of brucellosis-positive bovine cows. A total of 47 bovine ovarian follicular fluid aspirates from cows, positive in tests for brucellosis and from Brucella-positive herd, were submitted to PCR. The primers used in the PCR were specific to the genus Brucella (bcsp31 gene). All 47 bovine aspirates were negative for Brucella spp. 0.00% (95% CI: 0.00-4.00%). Our results demonstrated that Brucella spp. was absent in the ovarian follicular fluid from seropositive cows, which indicates that Brucella spp.-infected cows could be used for reproductive biotechnologies carried out with follicular aspirates. Future studies are needed to more precisely evaluate the feasibility and safety of using these oocytes from brucellosis-seropositive cows to transfer embryos to heifers/cows not infected by Brucella, aiming to produce calves free of the infection.

Machine learning for determining the stage of the estrous cycle in bitches

Reproductive biotechnologies, such as artificial insemination, are important tools in the reproduction of female dogs. Accurate determination of the specific stage of the estrous cycle is crucial for the successful application of these technologies.Vaginal cytology serves as a cost-effective and rapid diagnostic solution. However, itrelies on the analyzer’s expertise, making itsubject to human errors. Additionally, it may involve a prolonged duration between sample collection and result analysis.To minimize these limitations and streamline the diagnostic process,this studydeveloped software to automate the identification of the main phases of the estrous cycle that are important for artificial insemination. Eighteen vaginal cytology images were used, withsix images representing each of the phases studied (proestrus, estrus, and diestrus).Imageswere analyzed using the open source CellProfiller software, with subsequentclassification of the images using theTanagra software. Sensitivity and specificity valueswere determined for the proestrus, estrus, and diestrus phases, yielding results of 0.99, 0.86, 0.95, and 1, 0.95, 0.82, respectively.These findingsdemonstrate the model’scapacityfor correctly identifyingdifferent phases of the estrous cycle. The proposed model proved effective for the study’sobjective, and the authors suggest that it may be applicable to other economically important species.

Ecological Civilisation and Amphibian Sustainability through Reproduction Biotechnologies, Biobanking, and Conservation Breeding Programs (RBCs).

Intergenerational justice entitles the maximum retention of Earth's biodiversity. The 2022 United Nations COP 15, "Ecological Civilisation: Building a Shared Future for All Life on Earth", is committed to protecting 30% of Earth's terrestrial environments and, through COP 28, to mitigate the effects of the climate catastrophe on the biosphere. We focused this review on three core themes: the need and potential of reproduction biotechnologies, biobanks, and conservation breeding programs (RBCs) to satisfy sustainability goals; the technical state and current application of RBCs; and how to achieve the future potentials of RBCs in a rapidly evolving environmental and cultural landscape. RBCs include the hormonal stimulation of reproduction, the collection and storage of sperm and oocytes, and artificial fertilisation. Emerging technologies promise the perpetuation of species solely from biobanked biomaterials stored for perpetuity. Despite significant global declines and extinctions of amphibians, and predictions of a disastrous future for most biodiversity, practical support for amphibian RBCs remains limited mainly to a few limited projects in wealthy Western countries. We discuss the potential of amphibian RBCs to perpetuate amphibian diversity and prevent extinctions within multipolar geopolitical, cultural, and economic frameworks. We argue that a democratic, globally inclusive organisation is needed to focus RBCs on regions with the highest amphibian diversity. Prioritisation should include regional and international collaborations, community engagement, and support for RBC facilities ranging from zoos and other institutions to those of private carers. We tabulate a standard terminology for field programs associated with RBCs for publication and media consistency.