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OBJECTIVE: This study aimed to examine the behavior towards the acceptance of donor egg, donor sperm, and donor embryo of Northern Thai infertile couples, separated between men and women. METHODS: A cross-sectional study was conducted at the CMEx Fertility Center, Chiang Mai, Thailand. The questionnaires consisted of sociodemographic questions and the acceptance of couples toward donor egg, sperm and embryo. The couples filled in the answers separately. RESULTS: A total of 250 infertile couples were assessed. There were no differences in the acceptance rate of donor egg, sperm and embryo between the men and the women. Male acceptance rates were 25.6%, 18.8%, and 18.8%, respectively; while female acceptance rates were 24.4%, 18.4%, and 19.2%, respectively. Most couples (over 70%) concordantly rejected the donation program. Around 10% of couples had discordant answers. The concordance accepted for couples for donor egg, sperm and embryo was only 20%, 13.2%, and 14.8%. Older people and those who had been infertile for a longer period were significantly more likely to accept donation programs. CONCLUSIONS: There is no difference concerning the acceptance of donor gametes and embryo among men and women. Most participants reject the utilization of donor programs, the overall acceptance rate is relatively low. This may indicate the need for more adequate information and education for the community to enhance prevention programs rather than focus on the treatment with donor gametes or embryos.

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Over a reproductive cycle, the prevalence and intensity of degeneration of testicular follicles in Megapitaria squalida collected from the mining port of Santa Rosalia (a highly metal-polluted area), and San Lucas (a less polluted site), Gulf of California, Mexico, were evaluated. At San Lucas, most individuals had a typical testicular structure, and degeneration of testicular follicles was present in 9.5 % of spawning organisms. In contrast, at Santa Rosalia, 68 % of males, mainly in the ripe stage, had testicular degeneration (72 % severe intensity, mostly in medium and large-sized). Degeneration was characterized by intense hemocyte infiltration, identified as dense masses with numerous melanized cells in the follicle lumen. In both sites, males with testicular follicles degeneration had a lower condition index compared to males without degeneration. Degeneration of testicular follicles before spawning compromises and decreases the reproductive activity of M. squalida males at Santa Rosalia, which may ultimately affect the population sustainability.

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Abstract Introduction: Short-term gametes storage is an inexpensive and simple technique that allows the use of the same batch of eggs or sperm at different times, maximizing the application of research protocols and the use of gametes in production. Arbacia dufresnii is a sea urchin species with proven aquaculture potential and already used in the nutraceutical industry. Aging of its gametes is unknown and is a needed information to scale up the production. Objective: Determine the effect of male and female gamete aging on the fertilization success of Arbacia dufresnii. This will allow optimizing the use of gametes after collection decoupling spawning from fertilization. Methods: A. dufresnii individuals were induced to spawn and gametes were kept at 12 ± 1 °C throughout each bioassay. Sperm was separated into two treatments: activated sperm in seawater (AS), and dry sperm (DS). Two bioassays were made: Bioassay 1 evaluated the effect of time on fertility by performing fertilization tests at 0 h, 24 h, 48 h, 72 h, and 96 h after spawning. Bioassay 2 evaluated the contribution of each type of aged gamete on fertility, combining aged gametes (96 h) with fresh gametes (0 h). Results: Bioassay 1: the fertilization success obtained by combining eggs (E) with AS or DS presented important differences. While the fertilization success remained acceptable (greater than 50 %) for up to 72 h using ExDS, it only remained acceptable for up to 48 h using ExAS. Bioassay 2: acceptable fertilization success was found by combining aged E (96 h) with fresh sperm, or aged DS (96 h) with fresh E, but not using aged AS with fresh E. Conclusions: The findings of this work show that fertilization success in A. dufresnii gametes remains relatively unchanged for up to 48 h after spawning when combining ExAS, and for up to 72 h when combining ExDS. However, when combining aged E or aged DS with a fresh gamete, post-collection fertilization can be extended up to 96 h. In this work, the first steps have been taken to understand the conservation time of A. dufresnii gametes with minimum intervention.

Resumen Introducción: El almacenamiento de gametos a corto plazo es una técnica económica y sencilla que permite utilizar el mismo lote de óvulos o espermatozoides en diferentes momentos, maximizando la aplicación de protocolos de investigación y el uso de gametos en la producción. Arbacia dufresnii es una especie con probado potencial acuícola como fuente de gametos para la industria nutracéutica. Sin embargo, se desconoce el envejecimiento de sus gametos y es una información necesaria para escalar la producción. Objetivo: Determinar el efecto del envejecimiento de los gametos masculinos y femeninos en el éxito de la fecundación de Arbacia dufresnii con el fin de optimizar el aprovechamiento de los gametos después de la recolecta desincronizando el desove de la fecundación. Métodos: Se indujo el desove de individuos de A. dufresnii y los gametos se mantuvieron a 12 ± 1 °C durante cada bioensayo. El esperma se separó en dos tratamientos: esperma activado en agua de mar (AS) y esperma seco (DS). Se realizaron dos bioensayos: El Bioensayo 1 evaluó el efecto del tiempo sobre la fertilidad realizando pruebas de fecundación a las 0 h, 24 h, 48 h, 72 h y 96 h después del desove. El bioensayo 2 evaluó la contribución de cada tipo de gameta envejecida (96 h) sobre la fertilidad, combinando gametos envejecidas (96 h) con gametos frescas (0 h). Resultados: Bioensayo 1: el éxito de fecundación obtenido combinando huevos (E) con AS o DS presentó diferencias importantes. Si bien el éxito de la fecundación se mantuvo aceptable (más del 50 %) durante un máximo de 72 h con ExDS, solo permaneció aceptable hasta 48 h con ExAS. Bioensayo 2: se encontró un éxito de fecundación aceptable combinando E envejecidos (96 h) con esperma fresco, o DS envejecido (96 h) con E fresco (0 h), pero no usando AS envejecido con E fresco (0 h). Conclusiones: Los hallazgos de este trabajo muestran que el éxito de la fecundación en los gametos de A. dufresnii permanece relativamente sin cambios hasta 48 h después del desove cuando se combina ExAS, y hasta 72 h cuando se combina ExDS. Sin embargo, cuando se combina E envejecido o DS envejecido con un gameto fresco, el tiempo entre la recolección y la fecundación puede extenderse hasta 96 h. En este trabajo se han dado los primeros pasos para entender el tiempo de conservación de los gametos de A. dufresnii con mínima intervención.

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KEY MESSAGE: Unreduced megagametophytes via second-division restitution were confirmed through heterozygosity analysis, and four candidate physical centromeres of rubber were located for the first time. The evaluation of maternal heterozygosity restitution (MHR) is vital in identifying the mechanism of 2n gametogenesis and assessing the utilization value of 2n gametes. In this study, three full-sib triploid populations were employed to evaluate the MHR of 2n female gametes of rubber tree clone GT1 and to confirm their genetic derivation. The 2n female gametes of GT1 were derived from second-division restitution (SDR) and transmitted more than half of the parental heterozygosity. In addition, low recombination frequency markers were developed, and four candidate physical centromeres of rubber tree were located for the first time. The confirmation that 2n female gametes of rubber tree clone GT1 are derived from SDR provides insights into the molecular mechanisms of 2n gametogenesis. In addition, the identified centromere location will aid in the development of centromeric markers for the rapid identification of the 2n gametogenesis mechanism.

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The vitrification of ovarian follicles is a strategic tool that may contribute to advances in aquaculture and the conservation of many important species. Despite the difficulties inherent to the cryopreservation of oocytes, some successful protocols have been developed for different species, but little is known about the capacity of oocytes to develop after thawing. Therefore, the profiles of the reproductive pathway genes and fatty acid membrane composition during the initial stages of development were analyzed in fresh ovarian follicles and follicles after the vitrification process. There were differences in the expression of the hypothalamic-pituitary-gonad axis genes during the follicular development in the control group as well as in the vitrified group. Similarly, alterations in the composition of fatty acids were observed after vitrification. Despite this, many alterations were observed in the vitrified group; more than half of the stage III ovarian follicles were able to grow and mature in vitro. Therefore, the vitrification of ovarian follicles may impact them at molecular and membrane levels, but it does not compromise their capability for in vitro maturation, which indicates that the technique can be a strategic tool for aquaculture.

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OBJECTIVE: Infertility is one of the most significant reproductive health issues addressed with medically assisted procreation. This study looked into a potential correlation between the number of mature oocytes harvested and donor biological characteristics in order to propose an anti-Müllerian hormone (AMH) cutoff level to optimize the selection of candidates for gamete donation. METHODS: The donors were healthy women included in the Public Gamete Bank between 2011 and 2021. Their results can be used as a national indicator of fertility. RESULTS: We found that women with higher AMH levels had more antral follicles and oocytes harvested. As age increased, the number of oocytes harvested decreased. The suggested AMH cutoff level for successful donation was 1.12 ng/mL. CONCLUSIONS: The analysis of the reproductive health of Public Gamete Bank donors allows the standardization of AMH cutoff values at a national level, since the same laboratory techniques were employed consistently across medical centers. The study also allowed insight into the factors that compromise donation success. If adopted, a more rigorous selection of donor candidates would increase the success rate of egg donations.

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OBJECTIVE: Genetic counseling and carrier screening are part of the gamete donation process by healthy individuals. We aim to review the findings of genetic counseling and carrier screening of a cohort of candidates at our public gametes bank. METHODS: Thirty-four male and 64 female candidates had genetic counseling with a medical geneticist before donation. Of these, one female candidate voluntarily dropped-out. Thirty-four males and 63 females performed karyotype and screening for the more common pathogenic variants for CFTR-related cystic fibrosis and spinal muscular atrophy (SMN1) in the Portuguese population. In addition, all females also performed Fragile X expansion screening (FMR1). Thirty candidates with known or assumed African ancestry performed hemoglobinopathies screening. RESULTS: Six candidates were definitely or temporarily withheld from the donation process given their family or personal history that required further investigation. Of 97 candidates tested, 16.5% presented anomalous laboratory results (16/97): ten candidates were carriers for an autosomal recessive disorder - cystic fibrosis (5/97), sickle cell anemia (3/30), and spinal muscular atrophy (2/97). One female was an FMR1 pre-mutation carrier (1/63). One female candidate presented with triple X mosaicism: 47,XXX[2]/46,XX[50]. Two candidates presented with chromosomal instability of unknown origin. In one candidate, a mosaic for the Philadelphia chromosome was detected, revealing the diagnosis of chronic myeloid leukemia. CONCLUSIONS: From a cohort of 97 candidates, 21.7% had a family/personal history or an anomalous laboratory result that required additional genetic counseling, stressing the importance of performing pre-donation genetic counseling in this population.

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OBJECTIVE: Every individual has the right to a safe and satisfactory sex and reproductive life. Therefore, several countries have made efforts to make Reproductive Health resources available to their populations. However, few results have been published regarding the policies implemented in Portugal. This study looked into the reproductive health status of the Portuguese male population. METHODS: Sperm donor candidates from the Portuguese Public Gamete Bank registered between 2011 and 2018 were included in the study. Spermogram findings were evaluated with respect to sociodemographic and risk factors. RESULTS: This is the first study performed in this population. We found that sperm quality has decreased throughout the last nine years, and that spermatozoa progressive motility is inversely correlated with the body mass index. An association between drug use and decreased sperm pH was also found. CONCLUSIONS: Changes in sperm quality have important consequences in male fertility. Most of the identified causes of decreased sperm quality are modifiable factors and should therefore be addressed since an early age.

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Mammalian fertilization is a complex process involving a series of successive sperm-egg interaction steps mediated by different molecules and mechanisms. Studies carried out during the past 30 years, using a group of proteins named CRISP (Cysteine-RIch Secretory Proteins), have significantly contributed to elucidating the molecular mechanisms underlying mammalian gamete interaction. The CRISP family is composed of four members (i.e., CRISP1-4) in mammals, mainly expressed in the male tract, present in spermatozoa and exhibiting Ca2+ channel regulatory abilities. Biochemical, molecular and genetic approaches show that each CRISP protein participates in more than one stage of gamete interaction (i.e., cumulus penetration, sperm-ZP binding, ZP penetration, gamete fusion) by either ligand-receptor interactions or the regulation of several capacitation-associated events (i.e., protein tyrosine phosphorylation, acrosome reaction, hyperactivation, etc.) likely through their ability to regulate different sperm ion channels. Moreover, deletion of different numbers and combination of Crisp genes leading to the generation of single, double, triple and quadruple knockout mice showed that CRISP proteins are essential for male fertility and are involved not only in gamete interaction but also in previous and subsequent steps such as sperm transport within the female tract and early embryo development. Collectively, these observations reveal that CRISP have evolved to perform redundant as well as specialized functions and are organized in functional modules within the family that work through independent pathways and contribute distinctly to fertility success. Redundancy and compensation mechanisms within protein families are particularly important for spermatozoa which are transcriptionally and translationally inactive cells carrying numerous protein families, emphasizing the importance of generating multiple knockout models to unmask the true functional relevance of family proteins. Considering the high sequence and functional homology between rodent and human CRISP proteins, these observations will contribute to a better understanding and diagnosis of human infertility as well as the development of new contraceptive options.

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Abstract: 15. The anonymity of gamete donors in the context of medically-assisted reproduction techniques (ART) and the right of the offspring to know their genetic or biological parents' identity is a controversial and widely debated topic in the scientific literature. The positions on the issue in each country are different. Sometimes they are in opposition to each other even in countries with strong similarities, such as those in the European Union (EU), in the framework of shared ethical values. Although some countries still enshrine the rule of anonymity, there is an undeniable tendency to guarantee the right to know one's origins by creating relevant exceptions or abolishing donor anonymity status altogether. 16. This article offers ethical and legal considerations of whether the so-called 'right to be forgotten' (RTBF) could be extended to include gamete donors' right to remain anonymous. This perspective goes against the general trend, certainly in Europe, of recognizing that offspring born from donor gametes have a right to access information relating to their genetic progenitors. The novel addition is to question whether the General Data Protection Regulation (GDPR) might provide fertile ground for questioning this approach, and effectively support those jurisdictions where anonymity is still possible.

Resumen: 20. El anonimato de los donantes de gametos en el contexto de las técnicas de reproducción médicamente asistida (RM) y el derecho de la descendencia a conocer su identidad genética o biológica es un tema controvertido y ampliamente debatido en la literatura científica. Las posiciones sobre el tema en cada país son diferentes. A veces se oponen entre sí, incluso en países con fuertes similitudes, como los de la Unión Europea (UE), en el marco de valores éticos compartidos. Aunque algunos países siguen consagrando la norma del anonimato, es innegable la tendencia a garantizar el derecho a conocer el propio origen creando las excepciones pertinentes o suprimiendo por completo el estatus de anonimato del donante. 21. Este artículo ofrece consideraciones éticas y jurídicas sobre si el llamado "derecho al olvido" podría ampliarse para incluir el derecho de los donantes de gametos a permanecer en el anonimato. Esta opinión es contraria a la tendencia general, ciertamente en Europa, de reconocer que los hijos nacidos de gametos donados tienen derecho a acceder a la información relativa a sus padres genéticos. La nueva adición consiste en debatir si el Reglamento general de protección de datos (RGPD) podría proporcionar un terreno fértil para cuestionar este enfoque y apoyar efectivamente a las jurisdicciones en las que el anonimato sigue siendo posible.

Resumo 25. O anonimato dos dadores de gâmetas no contexto das técnicas de reprodução medicamente assistida (RMA) e o direito da descendência a conhecer a sua identidade genética ou biológica é um tema controverso e amplamente debatido na literatura científica. As posições sobre a questão em cada país são diferentes. Por vezes estão em oposição umas às outras, mesmo em países com fortes semelhanças, como os da União Europeia (UE), no quadro de valores éticos partilhados. Embora alguns países ainda consagrem a regra do anonimato, existe uma tendência inegável para garantir o direito de conhecer as suas origens, criando exceções relevantes ou abolindo completamente o estatuto de anonimato dos dadores. 26. Este artigo oferece considerações éticas e legais sobre se o chamado "direito ao esquecimento" poderia ser alargado para incluir o direito dos dadores de gâmetas a permanecerem anónimos. Esta perspetiva vai contra a tendência geral, certamente na Europa, de reconhecer que os descendentes nascidos de gâmetas doadas têm o direito de aceder à informação relacionada com os seus progenitores genéticos. O novo aditamento é debater se o Regulamento Geral de Proteção de Dados (RGPD) poderá fornecer um terreno fértil para questionar esta abordagem, e apoiar efetivamente as jurisdições onde o anonimato ainda é possível.

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