Abstract Overview and Aims: Delayed motherhood is a reality today, especially for women with higher levels of education. Elective oocyte cryopreservation reduces the risk of future infertility. Despite increasing discussions in the media and social networks, most women are not adequately informed about the fertility decline associated with age and the associated risk factors. The aim of our study was to access the knowledge and attitudes regarding motherhood, fertility, and elective oocyte cryopreservation among young female university students in Portugal. Study Design: Observational and descriptive study. Population: Female college students. Methods: Data were collected through an online Google Forms® survey during August and September 2023. Results: A total of 266 female students were included in the study, with an average age of 22.46 years (±4.2). When asked about their willingness to have children in the future, 65% (n=173) of participants responded “yes”, 24.8% (n=66) “maybe”, and 10.2% (n=27) “no”. The main reasons cited for “no” and “maybe” responses were career prioritization (68.8%), fear of childbirth (45.2%), fear of motherhood (43%) or socioeconomic issues (40%). Approximately 53.6% (n=121) of participants wished to have children after the age of 30 years. Concerns about future fertility were present in 69.2% (n=184) of participants, and 72.9% (n=194) were familiar with the concept of ovarian reserve, while 82.7% (n=220) were aware of fertility preservation methods. When asked about the possibility of opting for elective oocyte cryopreservation, only 14.3% (n=38) ruled out this possibility. Most young women (87,2%, n=232) wished to be more informed about the topic, and 87.6% (n=233) would like to discuss the issue with their gynaecologist. Conclusions: The majority of students considered motherhood, however their professional career was highlighted as the main priority for the next 10 years. Knowledge about the ovarian reserve decline and the effectiveness of oocyte cryopreservation was not satisfactory.

Resumo Introdução e Objetivos: A maternidade tardia é hoje uma realidade, principalmente para mulheres com níveis de escolaridade superiores. A criopreservação eletiva de ovócitos permite diminuir o risco de infertilidade futura. A maioria das mulheres não está adequadamente informada sobre o declínio da fertilidade associado à idade nem sobre os fatores de risco associados. O objetivo do estudo foi avaliar o conhecimento e atitudes em relação à maternidade, fertilidade e à criopreservação eletiva de ovócitos das jovens universitárias em Portugal. Tipo de estudo: Estudo observacional e descritivo. População: Estudantes universitárias do sexo feminino. Metodologia: Dados recolhidos através de inquérito online Google Forms® realizado durante os meses de agosto e setembro de 2023. Resultados: Foram incluídas no estudo 266 estudantes do sexo feminino, com uma média de idade de 22,46 anos (± 4,2). Quando inquiridas quanto à vontade de terem filhos no futuro, 65% (n=173) das participantes respondeu “sim”, 24,8% (n=66) “talvez” e 10,2% (n=27) “não”. O principal motivo apontado para as respostas “não” e “talvez” foi a priorização da carreira (68,8%). Cerca de 53,6% (n=121) das participantes apenas desejam ter filhos a partir dos 30 anos. Para 69,2% (n=184) a fertilidade futura é uma preocupação, 72,9% (n=194) está familiarizada com o conceito de reserva ovárica e 82,7% (n=220) com a existência de métodos de preservação da fertilidade. Quando questionadas quanto à possibilidade de recorrer à criopreservação eletiva de ovócitos, apenas 14,3% (n=38) exclui essa possibilidade. A maioria das jovens (87,2%, n=232) gostaria de estar mais informada sobre a temática e 87,6% (n=233) gostaria de ver discutido o tema com o seu médico. Conclusões: A maioria das estudantes equaciona a maternidade, contudo a carreira profissional é destacada como principal prioridade para os próximos 10 anos. O conhecimento sobre declínio da reserva ovárica e eficácia da criopreservação de ovócitos está ainda aquém do desejado.

ABSTRACT This experiment aimed to assess the effect of fetal calf serum (FCS) added to culture medium on day 5 to 7 after in vitro fertilization on production and subsequent cryotolerance of blastocysts from Ecuadorian Creole heifers. Immature cumulus-oocyte complexes (COCs) were collected by ovum pick-up (OPU) from 10 Creole heifers and from ovaries collected at abattoir in eight collection sessions. COCs were subjected to in vitro maturation (IVM), fertilization (IVF), and culture (IVC). On day 1 after IVF, presumptive embryos from each oocyte source (OPU [O] and Abattoir [A]) were allocated randomly in two groups according to whether 2.5% (v/v) FCS was added to culture medium on day 5 after IVF: 1) O-FCS+, 2) O-FCS-, 3) A-FCS+, and 4) A-FCS-. On day 7 after IVF, high quality embryos were vitrified. Cryotolerance of vitrified-warmed embryos was assessed according to blastocele re-expansion at 2 h and subsequent re-expansion and hatching at 24 and 48 h of re-incubation. Blastocysts rate on day 7 did not differ between oocyte source and FCS group. After vitrification/warming process, addition of FCS affected the re-expansion rate only at 2 h, irrespective of the oocyte source (p<0.05). Likewise, blastocysts hatching rate at 48 h of incubation was drastically affected only in OPU derived oocyte (p<0.01). Supplementation of FCS on day 5 after IVF did not improve blastocyst production and adversely affected the cryotolerance of in vitro bovine embryos derived from creole heifers raised on Ecuadorian Andean highlands.

RESUMEN El estudio tuvo como objetivo evaluar el efecto del suero fetal bovino (FCS) agregado al medio de cultivo los días 5 a 7 después de la fertilización in vitro sobre la producción y posterior criotolerancia de blastocistos de vaquillonas criollas ecuatorianas. Se recolectaron complejos cúmulo-ovocitos (COCs) inmaduros mediante aspiración intravaginal guiada por ultrasonido (OPU) de 10 vaquillonas criollas y de ovarios de matadero en ocho sesiones. Los COCs se sometieron a maduración in vitro (MIV), fecundación (FIV) y cultivo (IVC). El día 1 después de la FIV, los presuntos embriones de cada fuente de ovocitos (OPU [O] y Matadero [A]) se asignaron aleatoriamente en dos grupos según si se añadió FCS al 2.5% (v/v) al medio de cultivo el día 5 después de la FIV: 1) O-FCS+, 2) O-FCS-, 3) A-FCS+ y 4) A-FCS-. El día 7 después de la FIV se vitrificaron embriones de alta calidad. La criotolerancia de los embriones vitrificados-calentados se evaluó según la reexpansión del blastocele a las 2 h y la posterior reexpansión y eclosión a las 24 y 48 h de incubación. La tasa de blastocistos el día 7 no difirió entre la fuente de ovocitos y el grupo FCS. Después del proceso de vitrificación/ calentamiento, la adición de FCS solo afectó la tasa de reexpansión a las 2 h, independientemente de la fuente de ovocitos (p<0.05). Asimismo, la tasa de eclosión de los blastocistos a las 48 h de incubación se vio drásticamente afectada únicamente en los ovocitos derivados de OPU (p<0.01). En conclusión, la suplementación de FCS el día 5 después de la FIV no mejoró la producción de blastocistos y afectó negativamente la criotolerancia de embriones bovinos in vitro derivados de vaquillonas criollas criadas en los Andes ecuatorianos.

Abstract A total of 381 specimens of the tilapia Oreochromis mossambicus collected monthly from May 2017 to May 2018 in the Laguna de Los Patos, Cumaná, Venezuela, to evaluate reproductive parameters of this non-native species. Significant differences were found in relation to average height and weight between males and females, with the highest values in males. The sex ratio was 1:1.5 (males:females), which deviates significantly from the expected 1:1 ratio. The mean length of sexual maturity (Lm50) was 18.0 cm in females and 20.1 cm in males, reflecting that females mature at smaller sizes than males. The monthly variations of the gonadosomatic index (GSI) and the stages of sexual maturity show two reproductive peaks during the study, in October 2017 and April 2018, coinciding with the rainy and dry seasons in the region respectively. The condition factor (CF) showed significant differences between months, but not between sexes, with an average of 1.87 in females and 1.84 in males. The average absolute fecundity was 921 ± 604.6 eggs per fish, with a relative fecundity of 8.36 ± 3.09 eggs per gram of fish. Differences in oocyte size in mature females confirm that the species can spawn repeatedly over a period, which is considered an important factor for the establishment of tilapia in non-native environments. 38 201 Patos Cumaná Venezuela nonnative non native 11.5 115 1 1.5 5 1:1. malesfemales, malesfemales , (males:females) 11 1: Lm50 Lm (Lm50 180 18 0 18. 20 20. GSI (GSI study respectively CF (CF months sexes 187 87 1.8 184 84 92 6046 604 6 604. fish 836 8 36 8.3 309 3 09 3.0 period environments 11. 15 1. (males:females Lm5 (Lm5 2 9 60 83 8. 30 3. (Lm

Resumo Um total de 381 espécimes da tilápia Oreochromis mossambicus foram coletados mensalmente de maio de 2017 a maio de 2018, na Laguna de Los Patos, Cumaná, Venezuela, para avaliar parâmetros reprodutivos desta espécie não nativa. Foram encontradas diferenças significativas em relação às médias de altura e peso entre machos e fêmeas, sendo os maiores valores no sexo masculino. A razão sexual foi de 1:1,5 (machos:fêmeas), o que se desvia significativamente da proporção esperada de 1:1. O comprimento médio de maturação sexual (Lm50) foi de 18,0 cm nas fêmeas e 20,1 cm nos machos, refletindo que as fêmeas amadurecem em tamanhos menores que os machos. As variações mensais do índice gonadossomático (IGS) e dos estágios de maturidade sexual mostram dois picos reprodutivos durante o estudo, em outubro de 2017 e abril de 2018, coincidindo com as estações chuvosa e seca na região, respectivamente. O fator de condição (FC) apresentou diferenças significativas entre os meses, mas não entre os sexos, com média de 1,87 no sexo feminino e 1,84 no masculino. A fecundidade absoluta média foi de 921 ± 604,6 ovos por peixe, com fecundidade relativa de 8,36 ± 3,09 ovos por grama de peixe. Diferenças no tamanho dos ovócitos em fêmeas maduras confirmam que a espécie pode desovar repetidamente ao longo de um período, o que é considerado um fator importante para o estabelecimento de tilápias em ambientes não nativos. 38 201 2018 Patos Cumaná Venezuela nativa masculino 11,5 115 1 1,5 5 1:1, machosfêmeas, machosfêmeas , (machos:fêmeas) 11. 11 1. 1:1 Lm50 Lm (Lm50 180 18 0 18, 20 20, IGS (IGS estudo região respectivamente FC (FC meses sexos 187 87 1,8 184 84 92 6046 604 6 604, peixe 836 8 36 8,3 309 3 09 3,0 período nativos 11, 15 1, (machos:fêmeas 1: Lm5 (Lm5 2 9 60 83 8, 30 3, (Lm

Resumo O polvo com chifres Eledone cirrhosa, uma espécie de tamanho médio encontrada no Golfo Arábico ao largo da Arábia Saudita, foi coletado mensalmente no Golfo Arábico ao largo da cidade de Dammam durante o período de janeiro a dezembro de 2022. As amostras foram dissecadas e preparadas para exame por microscopia eletrônica. Durante a maturação genital, os túbulos seminíferos se estabelecem nos testículos, com espermatogônias ativas se dividindo. Espermatócitos 1 são observados no túbulo, seguidos de aumento de espermatogônias e espermatócitos em agosto. A espermiogênese começa com espermátides esféricas evoluindo para espermátides alongadas. Em setembro, as espermatogônias ativas, as divisões meióticas e o aumento da espermiogênese continuam. Os espermatozoides aparecem na Bolsa de Needham, indicando maturidade sexual. O ovário passa por vários estágios de desenvolvimento, com oócitos no estágio I em junho e julho, seguido pelo estágio II em outubro e novembro. No estágio IIIc, os cordões foliculares invadem o citoplasma do oócito, formando numerosas inclusões lipídicas e grânulos proteicos. O citoplasma contém cisternas de retículo endoplasmático e um Aparelho de Golgi pouco desenvolvido. O estágio IV ocorre em novembro, caracterizado pelo máximo desenvolvimento dos cordões foliculares e pelo início da vitelogênese. O ooplasma contém numerosas inclusões lipídicas, um sincício e células secretoras. A partir de dezembro, os oócitos do estágio V estão principalmente presentes, indicando a fase de atividade de secreção máxima. As plaquetas da gema acumulam-se no ooplasma do ovócito e o córion se forma na zona pelúcida. Em janeiro, os primeiros óvulos lisos são encontrados no ovário de alguns polvos e sua proporção aumenta continuamente. Este estudo teve como objetivo investigar a ação mitogênica da gonadotrofina bem como identificar os períodos de intensa multiplicação celular durante o ciclo sexual por meio de métodos citológicos. cirrhosa Saudita 2022 eletrônica genital testículos dividindo túbulo agosto alongadas setembro continuam Needham julho novembro IIIc oócito proteicos desenvolvido vitelogênese secretoras presentes máxima acumulamse acumulam pelúcida continuamente citológicos 202 20 2

Abstract The horned octopod Eledone cirrhosa, a medium-sized species found in Arabian Gulf off Saudi Arabia, was collected monthly from the Arabian Gulf off Dammam city during January to December 2022. Samples were dissected and prepared for examination using transmission electron microscopy. During genital maturation, seminiferous tubules are established in the testis, with active spermatogonia dividing. Spermatocytes 1 are observed in the tubule, followed by an increase in spermatogonia and spermatocytes in August. Spermiogenesis begins, with spherical spermatids evolving into elongated spermatids. In September, active spermatogonia, meiotic divisions, and increased spermiogenesis continue. Spermatozoa appear in Needham's pouch, indicating sexual maturity. The ovary undergoes various stages of development, with oocytes at stage I in June and July, followed by stage II in October and November. In stage III, follicular cords invade the oocyte's cytoplasm, forming numerous lipid inclusions and protein granules. The cytoplasm contains cisternae of endoplasmic reticulum and a poorly developed Golgi apparatus. Stage IV occurs in November, characterized by the maximum development of follicular cords and the beginning of vitellogenesis. The ooplasm contains numerous lipid inclusions, a syncytium, and secretory cells. From December, stage V oocytes are mainly present, indicating the activity phase of maximum secretion. Yolk platelets accumulate in the oocyte ooplasm, and chorion forms at the zona pellucida. In January, the first smooth eggs are found in some octopuses' ovary, with their proportion increasing steadily. This study aimed to investigate the mitogenic action of gonadotropin and identify the periods of intense cell multiplication during the sexual cycle using cytological methods. cirrhosa mediumsized medium sized Arabia 2022 microscopy maturation testis dividing tubule August begins September divisions continue Needhams Needham s pouch maturity July November III granules apparatus vitellogenesis syncytium cells present secretion pellucida octopuses steadily methods 202 20 2

Resumo O objetivo do presente estudo foi investigar o impacto sinérgico do α-tocoferol e do ácido α-linolênico (100 µM) na MIV e CIV de oócitos de búfala Nili Ravi. Os oócitos foram obtidos dos ovários de búfalos abatidos duas horas após o abate e levados ao laboratório. Complexos de oócitos cumulus de búfalo foram colocados aleatoriamente nos cinco grupos experimentais incluídos; GRUPO 1: Meio de maturação (MM) + 100 µM ALA (controle), GRUPO 2: MM + 100 µM ALA + 50 µM α-tocoferol, GRUPO 3: MM + 100 µM ALA + 100 µM α-tocoferol, GRUPO 4: MM + 100 µM ALA + 200 μM α-tocoferol e GRUPO 5: MM + 100 µM ALA + 300 μM α-tocoferol sob uma atmosfera de 5% de CO2 em ar a 38,5 °C por 22-24 h. A expansão cumulus e o estado de maturação nuclear foram determinados (Experimento 1). No experimento 2, os oócitos foram maturados como no experimento 1. Os oócitos maturados foram então fertilizados em meio de Tyrode's Albumina Lactato Piruvato (TALP) por cerca de 20 h e cultivados em meio de fluido oviductal sintético (SOF) para determinar o efeito do ácido α-linolênico (100 µM) e α-tocoferol em meio IVM em IVC de presumíveis zigotos. Para estudar o efeito do ácido α-linolênico (100 µM) em meio IVM e aumentar a concentração de α-tocoferol no meio de cultura no desenvolvimento inicial do embrião (Experimento 3), os presumíveis zigotos foram distribuídos aleatoriamente nos cinco grupos experimentais com concentração crescente de α-tocoferol em meios de cultura. Maior porcentagem de oócitos em estágio MII no experimento 1 (65,2 ± 2,0), embriões em estágio de mórula no experimento 2 (30,4 ± 1,5) e experimento 3 (22,2 ± 2,0) foram obtidos. No entanto, os resultados gerais para a expansão das células do cumulus, maturação do oócito para o estágio MII e desenvolvimento embrionário subsequente entre os tratamentos permanecem estatisticamente semelhantes (P> 0,05). A suplementação de α-tocoferol em meios de maturação com ácido α-linolênico e / ou em meios de cultura de embriões não aumentou ainda mais a maturação in vitro de oócitos ou a produção de embriões.

Abstract The objective of the current study was to investigate the synergistic impact of α-Tocopherol and α-Linolenic acid (100 µM) on IVM and IVC of Nili Ravi buffalo oocytes. Oocytes were obtained from the ovaries of slaughtered buffaloes within two hours after slaughter and brought to laboratory. Buffalo cumulus oocyte complexes were placed randomly in the five experimental groups included; GROUP 1: Maturation media (MM) + 100 µM ALA (control), GROUP 2: MM + 100 µM ALA + 50μM α-Tocopherol, GROUP 3: MM + 100 µM ALA + 100μM α-Tocopherol, GROUP 4: MM + 100 µM ALA + 200 μM α-Tocopherol and GROUP 5: MM + 100 µM ALA + 300 μM α-Tocopherol under an atmosphere of 5% CO2 in air at 38.5 °C for 22-24 h. Cumulus expansion and nuclear maturation status was determined (Experiment 1). In experiment 2, oocytes were matured as in experiment 1. The matured oocytes were then fertilized in Tyrode’s Albumin Lactate Pyruvate (TALP) medium for about 20 h and cultured in synthetic oviductal fluid (SOF) medium to determine effect of α-Linolenic acid (100 µM) and α-Tocopherol in IVM medium on IVC of presumptive zygotes. To study the effect of α-Linolenic acid (100 µM) in IVM media and increasing concentration of α-tocopherol in the culture media on early embryo development (Experiment 3), the presumptive zygotes were randomly distributed into the five experimental groups with increasing concentration of α-tocopherol in culture media. Higher percentage of MII stage oocytes in experiment 1(65.2±2.0), embryos at morula stage in experiment 2 (30.4±1.5) and experiment 3 (22.2±2.0) were obtained. However, overall results for cumulus cell expansion, maturation of oocyte to MII stage and subsequent embryo development among treatments remain statistically similar (P > 0.05). Supplementation of α-tocopherol in maturation media having α-Linolenic acid and/or in embryo culture media did not further enhance in vitro maturation of oocyte or embryo production.

Objective: Fertility preservation is a priority in oncology for female cancer patients. However, there is a lack of communication between infertility specialists and oncologists. This study aimed to evaluate infertility specialists’ perceptions and experiences regarding fertility preservation. Methods: Conduct an online survey to profile infertility specialists. Participants were infertility affiliated with the Brazilian Federation of Gynecology and Obstetrics Associations (FEBRASGO). The specialists received an online survey, which response rate were 30.9%, most of whom were in southern and southeastern. The survey consisted on 14 questions about the infertility specialists’ location, techniques in clinical practice, treatment successful rate, patients idea, etc. Results: The average experience in human reproduction were 15.5 ± 10.2 years (mean ± standard deviation, range 1-40). Among reproductive-aged female cancer patients recommended for fertility preservation, 60.3 ± 28.8% (range 10-100%) underwent preservation procedures. Main barriers were cost (41%), oncologists’ knowledge or acceptance (35%) and accessibility (9%). Most infertility specialists (58%) considered 40 years the limit for fertility preservation. Leukemia, lymphoma, breast and ovarian cancers were prioritized for fertility preservation, while lung, thyroid, gastric, and brain cancers were less relevant. Conclusion: This is the first Brazilian study about infertility specialists’ perceptions on oncology patients access to fertility preservation. These patients primarily receive treatment in the public health system, while infertility specialists mainly work in the private healthcare. This healthcare mode is currently fragmented, but integrating these experts is enhancing patient access to fertility preservation. Studies on this topic are still warranted. Objective However oncologists Methods FEBRASGO. FEBRASGO . (FEBRASGO) 309 30 9 30.9% southeastern 1 location practice idea etc Results 155 15 5 15. 102 10 2 10. mean deviation 140. 140 1-40) reproductiveaged reproductive aged 603 60 3 60. 288 28 8 28.8 10100% 10100 100% 100 10-100% procedures 41%, 41 41% , (41%) 35% 35 (35% 9%. 9% (9%) 58% 58 (58% 4 Leukemia lymphoma lung thyroid gastric relevant Conclusion system fragmented warranted (FEBRASGO 30.9 1-40 6 28. 1010 10-100 (41% (35 (9% (58 30. 1-4 101 10-10 (41 (3 (9 (5 1- 10-1 (4 ( 10-

Abstract: Aim This study evaluates the reproduction and fecundity of Leporinus lacustris in upper Paraná River floodplain environments with the objectives to (i) recognize the germ cells of folliculogenesis and oogenesis; (ii) characterize reproductive phases; (iii) investigate oocyte diameter and evaluate if this species has group synchronous or asynchronous oocyte development and if spawning is total or batch; (iv) estimate batch fecundity; (v) establish the relationship between fecundity/ovary weight, standard length, and total weight; (vi) verify the reproductive areas of this species in the upper Paraná River floodplain. Methods Sampling was carried out using gillnets in rivers, and open and closed lagoons, in March, June, September, and December 2013, 2014, 2015, 2016, 2018, and 2019 and March and September 2017. A total of 442 females were sampled and 111 subsampled, each of which had an ovary removed and fixed in Bouin solution. A subsample of 20 ovaries in the spawning capable phase were fixed in a 4% buffered formaldehyde solution to estimate the diameter of the oocytes and fecundity. The ovaries were dehydrated, embedded in historesin, cut, stained, and evaluated as regards folliculogenesis, oogenesis, development phases, and reproduction areas. Results The germ cells recorded were differentiated oogonia, primary and secondary growth oocytes, fully-grown and mature oocytes, and post-ovulatory follicles that permitted the recognition of the reproductive phases. Oocyte diameter information revealed that the species has group synchronous oocyte development, determinate fecundity and batch spawning, and eliminates an average of 38,490 oocytes. Conclusions The fact that Leporinus lacustris is reproductively successful in the Baía River and in the Fechada, Guaraná, Patos, Garças, Ventura and Ressaco do Pau Véio lagoons strengthens the importance of the areas of environmental protection of the islands and lowlands of the Paraná River, Ilha Grande National Park and Ivinheima River State Park. Abstract i (i oogenesis ii (ii phases iii (iii iv (iv v (v fecundityovary weight length vi (vi rivers June 2013 2014 2015 2016 2018 201 2017 44 11 subsampled 2 4 dehydrated historesin cut stained oogonia fullygrown fully grown postovulatory post ovulatory 38490 38 490 38,49 Fechada Guaraná Patos Garças 1 3849 3 49 38,4 384 38,

Resumo: Objetivo Este estudo avaliou a reprodução e fecundidade de L. lacustris em ambientes da planície de inundação do alto rio Paraná, com os objetivos de: (i) reconhecer as células germinativas da foliculogênese e oogênese; (ii) caracterizar as fases reprodutivas; (iii) investigar o diâmetro dos oócitos e avaliar se esta espécie tem o desenvolvimento dos oócitos sincrônico em grupo ou assincrônico e se a desova é total ou parcelada; (iv) estimar a fecundidade do lote; (v) estabelecer a relação entre fecundidade/peso do ovário, comprimento padrão e peso total e (vi) verificar as principais áreas de reprodução desta espécie na planície de inundação do alto rio Paraná. Métodos As amostragens foram realizadas em rios, lagoas abertas e fechadas no alto rio Paraná, com redes de espera nos meses de março, junho, setembro e dezembro de 2013, 2014, 2015, 2016, 2018 e 2019 e março e setembro de 2017. Foram amostradas 442 fêmeas e subamostradas 111, das quais foi retirada uma subamostra do ovário e fixada em solução de Bouin. Uma subamostra de 20 ovários na fase apta a desova foi fixada em solução de formaldeído tamponado a 4%, para posteriormente, estimar o diâmetro dos oócitos e fecundidade. Resultados As células germinativas registradas foram: oogônias diferenciadas, oócitos de crescimento primário e secundário, oócitos de crescimento completo e maduros, e folículos pós-ovulatórios que permitiram reconhecer as fases reprodutivas. As informações do diâmetro dos oócitos revelou que a espécie tem desenvolvimento dos oócitos sincrônico em grupo, fecundidade determinada e desova parcial, e elimina no período de desova em média 38.490 oócitos. Conclusões Leporinus lacustris reprodutivamente bem sucedido no rio Baía e nas lagoas Fechada, do Guaraná, dos Patos, das Garças, do Ventura e do Ressaco do Pau Véio reforçando a importância das áreas de proteção ambiental das ilhas e várzeas do rio Paraná, Parque Nacional da Ilha Grande e Parque Estadual do rio Ivinheima. Resumo L Paraná i (i oogênese ii (ii reprodutivas iii (iii parcelada iv (iv lote v (v fecundidadepeso vi (vi rios junho 2013 2014 2015 2016 201 2017 44 111 Bouin 2 4 4% posteriormente diferenciadas secundário maduros pósovulatórios pós ovulatórios parcial 38490 38 490 38.49 Fechada Guaraná Patos Garças Ivinheima 11 3849 3 49 38.4 1 384 38.

Abstract Background Basal energetic metabolism in sperm, particularly oxidative phosphorylation, is known to condition not only their oocyte fertilising ability, but also the subsequent embryo development. While the molecular pathways underlying these events still need to be elucidated, reactive oxygen species (ROS) could have a relevant role. We, therefore, aimed to describe the mechanisms through which mitochondrial activity can influence the first stages of embryo development. Results We first show that embryo development is tightly influenced by both intracellular ROS and mitochondrial activity. In addition, we depict that the inhibition of mitochondrial activity dramatically decreases intracellular ROS levels. Finally, we also demonstrate that the inhibition of mitochondrial respiration positively influences sperm DNA integrity, most likely because of the depletion of intracellular ROS formation. Conclusion Collectively, the data presented in this work reveals that impairment of early embryo development may result from the accumulation of sperm DNA damage caused by mitochondrial-derived ROS. phosphorylation ability elucidated (ROS role therefore addition levels Finally integrity formation Collectively mitochondrialderived derived

SUMMARY OBJECTIVE: The aim of this study was to determine the effect of anxiety and depression on pregnancy outcome in couples receiving in vitro fertilization treatment. METHODS: A total of 102 couples (102 females and 102 males) with unexplained infertility were included in the study. Personal Information Form was used to collect data, Case Follow-up Form to record the treatment process, and Spielberger’s State-Trait Anxiety Inventory and Beck Depression Inventory to measure the anxiety and depression levels of couples. Couples were measured twice: before treatment and on oocyte pickup day. RESULTS: There was no statistically significant difference between the anxiety and depression levels and oocyte count of women (p>0.05). There were statistically significant differences between State-Trait Anxiety Inventory level and sperm count and between BID level and sperm motility (p<0.05). There was no statistically significant difference between the anxiety and depression levels and pregnancy outcomes of women (p>0.05). CONCLUSION: Anxiety and depression had no effect on pregnancy outcome. More studies are needed to investigate the effect of anxiety and depression on pregnancy outcome in unexplained infertility. OBJECTIVE METHODS 10 (10 males data Followup Follow up process Spielbergers Spielberger s StateTrait State Trait twice day RESULTS p>0.05. p005 p p>0.05 . 0 05 (p>0.05) p<0.05. p<0.05 (p<0.05) CONCLUSION 1 (1 p00 p>0.0 (p>0.05 p<0.0 (p<0.05 ( p0 p>0. (p>0.0 p<0. (p<0.0 p>0 (p>0. p<0 (p<0. p> (p>0 p< (p<0 (p> (p< (p

Abstract The objective of this study was to evaluate the effect of three doses of equine chorionic gonadotropin (eCG) for ovarian superstimulation on ovarian response, follicular development and cumulus-oocyte complexes (COCs) collection in llamas. For this purpose, eighteen multiparous non-lactating adult (4–7 yo) female llamas with an average body condition of 2.8 (BCS 1–5) were submitted to a follicular ablation (FA) to induce a new follicular wave emergence. Two days after FA (Day 0), synchronized llamas were randomly allocated to three treatment groups (n = 6/group) and given 500, 750 and 1000 IU of eCG (Novormon®, Syntex, Buenos Aires, Argentina) per animal respectively to induce ovarian superstimulation. Transrectal ultrasonography were performed on Days 2, 4, and 6; and ovum pick up (OPU) was performed on Day 6. Data was evaluated by one-way analysis of variance (ANOVA), repeated measures ANOVA, and 2-tailed Chi-square. The average size (mm) of follicles was greater (p≤ 0.05) in the 1000 IU group compared to the other groups. There was a greater (p≤ 0.05) number of follicles ≥ 7 mm in the 1000 IU group compared to the 500 IU group. Number of COCs collected on Day 6 and the COC recovery rate were not different among groups. In conclusion, a single dose of 1000 IU of eCG induced the best ovarian response resulting in larger and greater number of follicles at the time of OPU. (eCG cumulusoocyte cumulus oocyte (COCs purpose nonlactating non lactating 4–7 47 4 (4– yo 28 2 8 2. BCS 1–5 15 1 5 (FA emergence 0, 0 , 0) n 6/group 6group 75 100 Novormon®, Novormon (Novormon® Syntex Aires Argentina OPU (OPU oneway one way ANOVA (ANOVA) 2tailed tailed Chisquare. Chisquare Chi square. square Chi-square (mm p≤ p (p 0.05 005 05 50 conclusion 4– (4 1– 10 Novormon® (Novormon (ANOVA 0.0 00 ( 0.

Abstract One of the crucial aspects to be considered for successful in vitro production (IVP) of embryos is the composition of the various media used throughout the stages of this reproductive biotechnology. The cell culture media employed should fulfill the metabolic requirements of both gametes during oocyte maturation and sperm development, as well as the embryo during its initial cell divisions. Most IVP protocols incorporate blood serum into the media composition as a source of hormones, proteins, growth factors, and nutrients. Numerous studies have suggested Platelet-Rich Plasma (PRP) as a substitute for fetal sera in cell culture, particularly for stem cells. Therefore, the objective of this study is to assess the potential use of PRP as a replacement for fetal bovine serum (FBS) during oocyte maturation for in vitro production of bovine embryos. During in vitro maturation (IVM), cumulus-oocyte complexes (COCs) were allocated into the following experimental groups: Group G1 (IVM medium with 5% PRP); Group G2 (MIV medium with 5% PRP and 5% SFB); Group G3 (MIV medium with 5% SFB); and Group G4 (MIV medium without either PRP or SFB). Subsequently, the cumulus-oocyte complexes were fertilized with semen from a single bull, and the resulting zygotes were cultured for seven days. Cleavage and blastocyst formation rates were assessed on days 2 and 7 of embryonic development, respectively. The quality of matured COCs was also evaluated by analyzing the gene expression of HSP70, an important protein associated with cellular stress. The results demonstrated that there were no significant differences among the experimental groups in terms of embryo production rates, both in the initial cleavage stages and blastocyst formation (except for the G4 group, which exhibited a lower blastocyst formation rate on D7, as expected). This indicates that PRP could be a cost-effective alternative to SFB in the IVP of embryos. (IVP biotechnology development divisions hormones proteins factors nutrients PlateletRich Platelet Rich (PRP cells Therefore FBS (FBS IVM, IVM , (IVM) cumulusoocyte cumulus (COCs G 5 PRP) MIV SFB) SFB. . Subsequently bull respectively HSP70 HSP stress except group D7 D expected. expected expected) costeffective cost effective HSP7

Abstract The subcortical maternal complex, which consists of maternal-effect genes, plays a crucial role in the development of oocytes and preimplantation embryo until the activation of the zygote genome. One such gene, known as peptidyl-arginine deiminase VI (Padi6), is involved in the oocyte maturation, fertilization and embryonic development. However, the precise function of Padi6 gene in buffalo is still unclear and requires further investigation. In this study, the sequence, mRNA and protein expression patterns of Padi6 gene were analyzed in oocytes, preimplantation embryos and somatic tissues of buffalo. The coding sequence of gene was successfully cloned and characterized. Real-time quantitative PCR results indicated an absence of Padi6 transcripts in somatic tissues. Notably, the expression levels of Padi6 in oocytes showed an increased from the germinal vesicle stage to metaphase II stage, followed by a rapid decrease during the morula and blastocyst stages. Immunofluorescence analysis confirmed these findings, revealing a noticeable decline in protein expression levels. Our research provides the initial comprehensive expression profile of Padi6 in buffalo oocytes and preimplantation embryos, serving as a solid foundation for further investigations into the functionality of maternal-effect genes in buffalo. complex maternaleffect effect genome peptidylarginine peptidyl arginine Padi6, Padi , (Padi6) maturation However investigation study characterized Realtime Real time Notably stages findings (Padi6 (Padi

Abstract This review aims to discuss the main factors involved in the development of early antral follicles until gonadotropin dependence. This follicular phase is characterized by intense proliferation of granulosa cells, formation of a fluid-filled cavity, morphological differentiation of cumulus cells, mural granulosa cells and recruitment of theca cells. The interaction between oocyte, granulosa and theca cells is crucial for follicular growth and hormone production. Growth factors produced by the oocyte, such as growth and differentiation factor-9 (GDF-9) and bone morphogenetic protein-15 (BMP-15), regulate granulosa cell proliferation and differentiation and antral cavity development, as well as stimulate the production of follicle-stimulating hormone (FSH) receptors in granulosa cells. In response to FSH, granulosa cells secrete C-type natriuretic peptide (CNP), which acts through its receptor to increase cyclic guanosine monophosphate (cGMP) production and consequently follicular development. Granulosa cells also produce insulin-like growth factor-1 (IGF-1) and increase aromatase enzyme activity, which results in greater sensitivity to gonadotropins and follicular steroidogenesis. The absence of IGF-1 signaling causes cessation of follicular growth at the early antral stage. Many other local factors are involved in the regulation of follicular development. Therefore, this review brings relevant data for a better understanding of the mechanisms involved in the control of early antral follicle growth, emphasizing the role of endocrine and paracrine factors, the oocyte-granulosa cell interaction and the processes of follicular atresia. The challenges for the establishment of efficient culture systems for in vitro growth of early antral follicles are also discussed. dependence fluidfilled fluid filled oocyte factor9 factor 9 factor- GDF9 GDF (GDF-9 protein15 protein 15 protein-1 BMP15, BMP15 BMP , (BMP-15) folliclestimulating stimulating FSH (FSH Ctype C type CNP, CNP (CNP) cGMP (cGMP insulinlike insulin like factor1 1 IGF1 IGF (IGF-1 activity steroidogenesis IGF- stage Therefore oocytegranulosa atresia discussed (GDF- protein1 protein- BMP1 (BMP-15 (CNP (IGF- (GDF (BMP-1 (IGF (BMP- (BMP

Resumo Esta revisão tem como objetivo discutir os principais fatores envolvidos no desenvolvimento de folículos antrais iniciais até a dependência de gonadotrofinas. Essa fase folicular é caracterizada por intensa proliferação de células da granulosa, formação de uma cavidade preenchida por líquido, diferenciação morfológica das células do cumulus, células da granulosa murais e recrutamento de células da teca. A interação entre oócito, células da granulosa e da teca é determinante para o crescimento folicular e produção hormonal. Fatores de crescimento produzidos pelo oócito, fator de crescimento e diferenciação-9 (GDF-9) e proteína morfogenética óssea-15 (BMP-15), regulam a proliferação e diferenciação de células da granulosa, e o desenvolvimento da cavidade antral, bem como estimulam a produção de receptores do hormônio folículo estimulante (FSH) nas células da granulosa. Em resposta ao FSH, as células da granulosa secretam o peptídeo natriurético tipo C (CNP), que atua através de seu receptor para aumentar a produção de monofosfato de guanosina cíclico (GMPc) e consequentemente o desenvolvimento folicular. As células da granulosa também produzem o fator de crescimento semelhante à insulina 1 (IGF-1) e aumentam a atividade da enzima aromatase, o que resulta em maior sensibilidade às gonadotrofinas e esteroidogênese folicular. A ausência de sinalização do IGF-1 causa cessação do crescimento folicular no início do estágio antral. Muitos outros fatores locais estão envolvidos na regulação do desenvolvimento folicular. Por tanto essa revisão traz dados relevantes para uma melhor compreensão dos mecanismos envolvidos no controle do crescimento de folículos antrais iniciais, enfatizando o papel dos fatores endócrinos e parácrinos, a interação oócito-células da granulosa e os processos de atresia folicular. Os desafios para o estabelecimento de sistemas de cultivo eficientes para o crescimento in vitro de folículos antrais iniciais também são discutidos. líquido cumulus oócito hormonal diferenciação9 9 diferenciação- GDF9 GDF (GDF-9 óssea15 óssea 15 óssea-1 BMP15, BMP15 BMP , (BMP-15) antral FSH (FSH CNP, CNP (CNP) GMPc (GMPc IGF1 IGF (IGF-1 aromatase IGF- parácrinos oócitocélulas discutidos (GDF- óssea1 óssea- BMP1 (BMP-15 (CNP (IGF- (GDF (BMP-1 (IGF (BMP- (BMP

Abstract Follicular dynamics in dairy sheep is poorly understood. This study assessed the follicular dynamics of dairy sheep within and out of the breeding season by observing the effect of two progesterone concentrations (60 and 120 mg) used in vaginal pessaries in pre-synchronization. The experiment was conducted in April/May and October/November using 40 healthy multiparous dairy ewes predominantly of the Lacaune breed, which underwent daily transrectal ultrasound examinations. The number of ovulations (1.35 vs 1.05), ovulatory follicle diameter (mm) (5.97 vs 5.05), length of the luteal phase (11 vs 9.14 days), and length of the estrous cycle (16.83 vs 16.6 days) were assessed within and out of the breeding season, respectively. The sheep out of the breeding season did not present a corpus luteum at the time of insertion of the vaginal pessary, characterizing anestrus. The ovulatory follicle had a larger diameter in the breeding season even with the shorter length of the follicular phase, suggesting higher viability and quality of the ovulated oocyte. The application of exogenous progesterone extends the luteal phase in the breeding season (11 vs 9.14 days). The anestrus observed out of season suggests the need for a higher progesterone concentration in reproductive protocols. Therefore, breeding protocols can be adapted for different seasons, suggesting the use of vaginal pessaries with a higher progesterone concentration (120 mg) out of the breeding season. The data allowed the characterization of follicular dynamics in dairy sheep, demonstrating their differences in each season based on the use of different progesterone doses in pre-synchronization. understood 60 (6 12 mg presynchronization. presynchronization pre synchronization. synchronization pre-synchronization AprilMay April May OctoberNovember October November 4 breed examinations 1.35 135 1 35 (1.3 1.05, 105 1.05 , 05 1.05) mm (mm 5.97 597 5 97 (5.9 5.05, 505 5.05 5.05) 11 (1 914 9 14 9.1 days, days 16.83 1683 16 83 (16.8 166 6 16. respectively pessary oocyte days. . Therefore seasons (12 ( 1.3 13 3 (1. 10 1.0 0 5.9 59 (5. 50 5.0 91 9. 16.8 168 8 (16. 1. 5. (5 (16

Resumo A dinâmica folicular em ovelhas leiteiras é pouco compreendida. Neste trabalho foi avaliado a dinâmica folicular de ovelhas leiteiras, dentro e fora da estação reprodutiva observando o efeito de duas concentrações de progesterona (60 e 120mg) utilizadas em pessários vaginais na pré-sincronização. O experimento foi realizado nos meses de abril/maio e outubro/novembro, utilizando 40 ovelhas multíparas leiteiras hígidas, predominantemente da raça Lacaune, que foram submetidas a exames ultrassonográficos diários por via transretal. Na estação reprodutiva foram avaliados o número de ovulações (1,35 vs 1,05), o diâmetro(mm) do folículo ovulatório (5,97 vs 5,05), a duração da fase lútea (11 vs 9,14 dias) e a duração do ciclo estral (16,83 vs 16,6 dias) dentro da estação em comparação com fora da estação reprodutiva respectivamente. Fora da estação as ovelhas não apresentaram corpo lúteo no momento da inserção do pessário vaginal, caracterizando anestro. Mesmo com menor duração da fase folicular, o folículo ovulatório apresentou maior diâmetro na estação reprodutiva, sugerindo maior viabilidade e qualidade do oócito ovulado. A aplicação de progesterona exógena amplia a fase lútea na estação reprodutiva (11 vs 9,14 dias). O anestro observado fora de época sugere a necessidade de maior concentração de progesterona nos protocolos reprodutivos. Desta forma, é possível adaptar protocolos reprodutivos para diferentes épocas, sugerindo o uso de pessários vaginais com maior concentração de progesterona (120 mg) fora da estação reprodutiva. Os dados obtidos permitiram caracterizar a dinâmica folicular em ovelhas leiteiras, demonstrando suas diferenças em cada estação a partir do uso de diferentes doses de progesterona na pré-sincronização compreendida 60 (6 120mg mg présincronização. présincronização pré sincronização. sincronização abrilmaio abril maio outubronovembro outubro novembro outubro/novembro 4 hígidas Lacaune transretal 1,35 135 1 35 (1,3 1,05, 105 1,05 , 05 1,05) diâmetromm mm diâmetro(mm 5,97 597 5 97 (5,9 5,05, 505 5,05 5,05) 11 (1 914 9 14 9,1 dias 16,83 1683 16 83 (16,8 166 6 16, respectivamente vaginal ovulado dias. . forma épocas 120 (12 ( 1,3 13 3 (1, 10 1,0 0 5,9 59 (5, 50 5,0 91 9, 16,8 168 8 (16, 12 1, 5, (5 (16

Abstract Lipid metabolism is essential for ensuring oocyte maturation and embryo development. β-Oxidized fatty acids (FA) are a potent source of energy for cells, particularly for bovine somatic follicular cells. Superstimulatory protocols using follicle stimulating hormone (FSH) or FSH combined with equine chorionic gonadotropin (eCG) are capable of stimulating the follicular microenvironment and drive the expression of biomarker genes associated with lipid metabolism in the cumulus-oocyte complex (COC) for better embryo development. In this study, we assesed the effects of FSH and FSH/eCG protocols on the expression of genes related to lipid metabolism in bovine granulosa cells (GCs). Further, we measured triglyceride levels in follicular fluid (FF) obtained from both superstimulatd and non-superstimulated cows (synchronized cows). In summary, superstimulation with gonadotropins maintained the TG levels in bovine FF and ensured GCs mRNA abundance of ACSL1, ACSL3, ACSL6, SCD, ELOVL5, ELOVL6, FASN, FADS2, and SREBP1. We, however, found the abundance of CPTIB mRNA to be lower in GCs obtained from cows subjected to FSH/eCG protocols than synchronized cows. In conclusion, the findings of this study showed that ovarian superstimulation around the preovulatory phase has a mild impact on the lipid metabolism in GCs. development βOxidized β Oxidized FA (FA (FSH eCG (eCG cumulusoocyte cumulus COC (COC FSHeCG . (GCs) Further (FF nonsuperstimulated non superstimulated cows) summary ACSL1 ACSL ACSL3 ACSL6 SCD ELOVL5 ELOVL ELOVL6 FASN FADS2 FADS SREBP1 SREBP We however conclusion (GCs