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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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La reproducción asistida con donación ha venido a interrogar y a ampliar la noción de parentesco. La hegemonía del modelo biogenético dificulta que los padres de estas familias incorporen con tranquilidad la donación en su gestación. Desde los inicios de la técnica los padres han optado por no contar a su decendencia la historia de su concepción, muchas veces por miedo a que se deslegitimara su relación filial. Con el tiempo, se ha observado una apertura hacia la comunicación. Este cambio va de la mano con el desarrollo de la investigación en el área, que comenzó preguntándose por el bienestar psicológico de los nacidos por donación, por los aspectos relacionales de las familias concebidas de este modo e interrogando los beneficios de la comunicación de orígenes. Luego, investigando las percepciones en relación con su origen de las personas nacidas por donación. Finalmente, se ha publicado acerca la imposibilidad del anonimato en la era de los exámenes genéticos directos al consumidor. El modo de entender la reproducción con donante ha evolucionado, de un modelo médico en el que el foco es el lograr un embarazo saludable, a un modelo de formación de familia, en el que la motivación es contribuir a formar familias de buen funcionamiento. Esto tiene implicancias en la consejería a quienes participan del proceso, y también en los lineamientos que las sociedades científicas y comités de ética sugieren a los programas de reproducción con donante.

Gamete donation has come to question and broaden the notion of kinship. The hegemony of an biogenetic model has been an obstacle for parents of these families to incorporate donation into their history. From the beginning of the technique the parents chose not to tell their descent the history of their conception, often for fear that their filial relationship would be delegitimized. Over time, an openness to disclosure has been observed. This change goes hand in hand with the development of research in the area, which began by asking about the psychological well-being of those born by donation. Then investigating the relational aspects of families conceived in this way and questioning the benefits of disclosure. Subsequently, it was possible to study the perceptions regarding their origin of people born by donation. Finally, the impossibility of anonymity in the era of direct consumer genetic testing has been published. The way of understanding donor reproduction has evolved, from a medical model in focus is to achieve a healthy pregnancy, a family formation model, in which the motivation is to contribute to forming well-functioning families. This has implications in counseling to those who participate in the process, and in the guidelines that scientific societies and ethics committees affect donor reproduction programs.

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Populations of gray brocket deer (Mazama gouazoubira) are declining; yet, knowledge on the reproductive biology of this species remains limited. Therefore, this study aimed to describe morphology, viability, membrane integrity, mitochondrial activity, morphometry, micromorphology, and ultrastructure of the gray brocket deer sperm. Three adult male gray brocket deer were used in the study. Semen collection was performed using electroejaculation. Semen were analyzed by evaluating pH, motilities, vigor, mass movement, volume, concentration, viability, membrane integrity, mitochondrial activity, morphology, and morphometry. Micromorphology and ultrastructure of sperm were analyzed using scanning and transmission electron microscopy (SEM and TEM), respectively. There was no significant difference among males regarding on pH, motilities, vigor, mass movement, volume, concentration, viability. High values for membrane integrity, mitochondrial activity, and normal sperm were observed. The most frequent defects were simple bent tail and bowed midpiece. The head length, and width, midpiece, and tail length were 8.5, 4.4, 11.5, and 41.3 µm, respectively. SEM sperm showed paddle-shaped heads, with apical ridge and serrated band on the equatorial segment. TEM revealed the nucleus, acrosome, plasma membrane, mitochondria sheath, proximal centrioles, segmented columns, axoneme, outer dense fibers, and fibrous sheath. SEM and TEM showed the presence of some abnormalities. These results are expected to provide baseline values of diverse semen parameters, contributing toward the development of reproductive biotechnologies for gray brocket deer and, other deer species at risk of extinction.

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In vitro gamete derivation based on differentiation of germ cells (GC) from stem cells has emerged as a potential new strategy for the treatment of male infertility. This technology also has potential applications in animal reproduction as an alternative method for dissemination of elite animal genetics, production of transgenic animals, and conservation of endangered species. Mesenchymal stem cells (MSC) are multipotent progenitor cells defined by their ability to differentiate into mesodermal lineages. Under the effect of selected bioactive factors, MSC upregulate expression of pluripotent and GC specific-markers revealing their potential for GC differentiation. In addition to the effect of trophic factors, cell-to-cell interaction with Sertoli cells (SC) may be required to guide the sequential differentiation of MSC into GC. Thus, the aim of the present study was to investigate the effect of coculture with SC on the potential for in vitro GC differentiation of bovine fetal MSC (bfMSC) derived from bone marrow (BM-MSC) and adipose tissue (AT-MSC). bfMSC were isolated from male bovine fetuses and SC were collected from adult bull testes. The effect of SC interaction with BM-MSC or AT-MSC was analyzed on the expression of pluripotent factors OCT4 and NANOG, GC genes FRAGILLIS, STELLA and VASA and male GC markers DAZL, PIWIL2, STRA8 and SCP3 at Day 14 of coculture. Flow cytometry analyses detected that the majority (95,5% ±â€¯2.5; P < 0.05) of the isolated population of SC cultures were positive for SC-specific marker WT1. Levels of mRNA of WT1 in BM-MSC and AT-MSC were lower (P < 0.05) compared to SC; whereas, WT1 expression was not detected in bovine fetal fibroblasts (FB). Cocultures of BM-MSC and AT-MSC with SC had higher (P < 0.05) OCT4 mRNA levels compared to monocultures of BM-MSC, AT-MSC and SC. Moreover, cocultures of BM-MSC with SC had higher (P < 0.05) proportion of cells positive for Oct4 and Nanog compared to monocultures of BM-MSC and SC. Levels of mRNA of DAZL, PIWIL2 and SCP3 were upregulated in cocultures of AT-MSC with SC compared to monocultures of AT-MSC and SC. Accordingly, the proportion of cells positive for Dazl were higher (P < 0.05) in cocultures of AT-MSC with SC compared to monocultures of AT-MSC and SC. Changes in gene expression profiles during coculture of SC with AT-MSC suggest that cell-to-cell interaction or bioactive factors provided by SC may induce progression of AT-MSC into early stages of GC differentiation.

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The use of donated gametes and embryos in assisted reproduction techniques (ART) makes it necessary to examine interests that involve relevant ethical and legal considerations, which include the autonomy and privacy rights of the intended parents, donors' right to privacy and the right of the minors to know their genetic origin. This article presents arguments to consider policies of more openness to obtain information from donors in order to protect the child's best interest in knowing his/her genetic origins. It concludes with the situation in Mexico, where ART has been carried out with donated gametes since several years ago; however, due to the absence of regulations to control these procedures, each establishment imposes its own criteria for the operation of its programs.

El uso de gametos y embriones donados en técnicas de reproducción humana asistida hace necesario examinar intereses que implican consideraciones éticas y jurídicas relevantes, y estos incluyen los derechos de autonomía y privacidad de los padres de intención, el derecho a la privacidad de los donantes y el derecho de los menores a conocer su origen genético. En este artículo se exponen argumentos para considerar políticas de mayor apertura para la obtención de información de los donantes en aras de proteger el interés superior del niño y la niña a conocer sus orígenes genéticos. Concluye con la problemática en México, donde desde hace varios años se realizan técnicas de reproducción humana asistida con gametos donados, sin embargo, debido a la ausencia de una normatividad que regule estos procedimientos, cada establecimiento impone sus criterios para el funcionamiento de sus programas.

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The use of donated gametes and embryos in assisted reproduction techniques (ART) makes it necessary to examine interests that involve relevant ethical and legal considerations, which include the autonomy and privacy rights of the intended parents, donors’ right to privacy and the right of the minors to know their genetic origin. This article presents arguments to consider policies of more openness to obtain information from donors in order to protect the child’s best interest in knowing his/her genetic origins. It concludes with the situation in Mexico, where ART has been carried out with donated gametes since several years ago; however, due to the absence of regulations to control these procedures, each establishment imposes its own criteria for the operation of its programs.

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Perumytilus purpuratus is a marine mussel considered a bioengineer species with a broad distribution in the Pacific and Atlantic coast of South America. Studies have shown two geographically and genetically differentiated subpopulations at molecular level and in sperm morphological traits. To open avenues for molecular research on P. purpuratus, a global de novo transcriptome from gonadal tissue of mature males was sequenced using the Illumina platform. From a total of 126.38 million reads, 37,765 transcripts were successfully annotated. BUSCO analysis determined a level of 89% completeness for the assembled transcriptome. The functional gene ontology (GO) annotation indicated that, in terms of abundance, the transcripts related with molecular function were the most represented, followed by those related with biological process and cellular components. Additionally, a subset of GO annotations generated using the "sperm" term resulted in a total of 1,294 sequences where the biological process category was the more represented, with transcripts strongly associated to sperm-processes required for fertilization, and with processes where the sperm-egg interaction could be implicated. Our work will contribute to the evolutionary understanding of the molecular mechanisms related to tissue-specific functions. This work reports the first male gonad transcriptome for the mussel P. purpuratus, generating a useful transcriptomic resource for this species and other closely related mytilids.

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The present study aimed to evaluate the activation characteristics of the lebranche mullet spermatozoa in natura and diluted with CF-HBSS for 96h at 4±2°C. The semen was collected from eight wild fish in Florianópolis – SC (Brazil) (27°S) in May, during reproductive migration. Three pools of semen were divided into two treatments: in natura and diluted with CF-HBSS 1:3. The semen was activated with seawater (salinity of 34, pH 8.7 and 4±2°C) to determine: motility time, motility rate and sperm cell membrane integrity. Dilution with CF-HBSS 1:3 increased motility time of diluted semen (15- 20% for 6, 12 and 18h). Motility rate was equal to fresh semen for up to 24h of refrigeration and it was 30% higher than in natura semen at 12, 18 and 24h. Cell membrane integrity was maintained in fresh semen up to 6h, and it was 15-20% higher than in natura semen from 6 to 48h (p < 0.05). The use of CF-HBSS as diluent in the ratio 1:3 is recommended for semen refrigeration of lebranche mullet.

O objetivo do presente estudo foi avaliar em tainha as características de ativação dos espermatozoides in natura e diluídos com CF-HBSS mantidos por 96h a 4±2°C. O sêmen foi coletado de oito peixes selvagens em Florianópolis – SC (27°S) em maio, durante a migração reprodutiva. Três pool de sêmen foram divididos em dois tratamentos: in natura e diluído com CF-HBSS 1:3. O sêmen foi ativado com água oceânica (34 de salinidade, pH 8,7 e 4±2°C) para determinar: tempo de motilidade, taxa de ativação espermática e integridade da membrana celular. A diluição com CF-HBSS 1:3 propiciou o aumento no tempo de motilidade do sêmen diluído (15-20% para 6, 12 e 18h); manteve a taxa de motilidade igual ao sêmen fresco por até 24h de refrigeração e foi 30% maior do que em in natura em 12, 18 e 24h; manteve também a integridade da membrana celular igual ao sêmen fresco por 6h e 15-20% maior que em sêmen in natura (p < 0.05) para o período de 6 a 48h. O uso do CF-HBSS como diluidor na proporção 1:3 é recomendado para a refrigeração do sêmen da tainha.

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The present study aimed to evaluate the activation characteristics of the lebranche mullet spermatozoa in natura and diluted with CF-HBSS for 96h at 4±2°C. The semen was collected from eight wild fish in Florianópolis SC (Brazil) (27°S) in May, during reproductive migration. Three pools of semen were divided into two treatments: in natura and diluted with CF-HBSS 1:3. The semen was activated with seawater (salinity of 34, pH 8.7 and 4±2°C) to determine: motility time, motility rate and sperm cell membrane integrity. Dilution with CF-HBSS 1:3 increased motility time of diluted semen (15- 20% for 6, 12 and 18h). Motility rate was equal to fresh semen for up to 24h of refrigeration and it was 30% higher than in natura semen at 12, 18 and 24h. Cell membrane integrity was maintained in fresh semen up to 6h, and it was 15-20% higher than in natura semen from 6 to 48h (p < 0.05). The use of CF-HBSS as diluent in the ratio 1:3 is recommended for semen refrigeration of lebranche mullet.(AU)

O objetivo do presente estudo foi avaliar em tainha as características de ativação dos espermatozoides in natura e diluídos com CF-HBSS mantidos por 96h a 4±2°C. O sêmen foi coletado de oito peixes selvagens em Florianópolis SC (27°S) em maio, durante a migração reprodutiva. Três pool de sêmen foram divididos em dois tratamentos: in natura e diluído com CF-HBSS 1:3. O sêmen foi ativado com água oceânica (34 de salinidade, pH 8,7 e 4±2°C) para determinar: tempo de motilidade, taxa de ativação espermática e integridade da membrana celular. A diluição com CF-HBSS 1:3 propiciou o aumento no tempo de motilidade do sêmen diluído (15-20% para 6, 12 e 18h); manteve a taxa de motilidade igual ao sêmen fresco por até 24h de refrigeração e foi 30% maior do que em in natura em 12, 18 e 24h; manteve também a integridade da membrana celular igual ao sêmen fresco por 6h e 15-20% maior que em sêmen in natura (p < 0.05) para o período de 6 a 48h. O uso do CF-HBSS como diluidor na proporção 1:3 é recomendado para a refrigeração do sêmen da tainha.(AU)

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BACKGROUND: Bovine babesiosis is a tick-borne disease caused by the protozoan parasites of the genus Babesia. In their host vector, Babesia spp. undergo sexual reproduction. Therefore, the development of sexual stages and the subsequent formation of the zygote are essential for the parasite to invade the intestinal cells of the vector tick and continue its life-cycle. HAP2/GCS1 is a protein identified in plants, protozoan parasites and other organisms that has an important role during membrane fusion in fertilization processes. The identification and characterization of HAP-2 protein in Babesia would be very significant to understand the biology of the parasite and to develop a transmission-blocking vaccine in the future. RESULTS: To isolate and sequence the hap2 gene DNA from an infected bovine with Babesia bigemina was purified. The hap2 gene was amplified, cloned and sequenced. The sequences of hap2 from four geographically different strains showed high conservation at the amino acid level, including the typical structure with a signal peptide and the HAP2/GSC domain. Antisera anti-HAP2 against the conserved extracellular region of the HAP2 amino acid sequence were obtained from rabbits. The expression of hap2 in the host and vector tissues was analyzed by using semi-quantitative RT-PCR, and the protein was examined by western blot and immunofluorescence. Based on the RT-PCR and WB results, HAP2 is expressed in both, sexual stages induced in vitro, and in infected ticks as well. We did not detect any expression in asexual erythrocytic stages of B. bigemina, relevantly anti-HAP2 specific antibodies were able to block zygotes formation in vitro. CONCLUSION: Babesia bigemina HAP2 is expressed only in tick-infecting stages, and specific antibodies block zygote formation. Further studies regarding the function of HAP2 during tick infection may provide new insights into the molecular mechanisms of sexual reproduction of the parasite.

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The long-term impacts of recent marked increases in the incidence and extent of hypoxia (dissolved oxygen <2 mg/L) in coastal regions worldwide on fisheries and ecosystems are unknown. Reproductive impairment was investigated in Atlantic croaker collected in 2010 from the extensive coastal hypoxic region in the northern Gulf of Mexico. Potential fecundity was significantly lower in croaker collected throughout the ~20,000 km(2) hypoxic region than in croaker from normoxic sites. In vitro bioassays of gamete viability showed reductions in oocyte maturation and sperm motility in croaker collected from the hypoxic sites in response to reproductive hormones which were accompanied by decreases in gonadal levels of membrane progestin receptor alpha, the receptor regulating these processes. The finding that environmental hypoxia exposure reduces oocyte viability in addition to decreasing oocyte production in croaker suggests that fecundity estimates need to be adjusted to account for the decrease in oocyte maturation.

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