ABSTRACT Introduction Obstructive azoospermia occurs when there is a blockage in the male reproductive tract, leading to a complete absence of sperm in the ejaculate. It constitutes around 40% of all cases of azoospermia (1, 2). Blockages in the male reproductive tract can arise from either congenital or acquired factors, affecting various segments such as the epididymis, vas deferens, and ejaculatory ducts (3). Examples of congenital causes encompass conditions like congenital bilateral absence of the vas deferens and unexplained epididymal blockages (4). Acquired instances of obstructive azoospermia may result from factors like vasectomy, infections, trauma, or unintentional injuries caused by medical procedures (5). This complex condition affecting male fertility, presents two main treatment options: microsurgical reconstruction and surgical extraction of sperm followed by in vitro fertilization (IVF). Microsurgical reconstruction proves to be the most cost-effective option for treating obstructive azoospermia when compared with assisted reproductive techniques (6, 7). However, success rates of reconstruction defined by patency are as high as 99% for vasovasostomy (VV) but decline to around 65% if vasoepididymostomy (VE) is required (8, 9). Thus, continued refinement in technique is necessary in order to attempt to improve patency for patients undergoing VE. In this video, we show a comprehensive demonstration of microsurgical VE, highlighting the innovative epididymal occlusion stitch. The goal of this innovative surgical technique is to improve outcomes for VE. Materials and Methods The patient is a 39-year-old male diagnosed with obstructive azoospermia who presents for surgical reconstruction via VE. His partner is a 37-years-old female with regular menstrual cycles. The comprehensive clinical data encompasses a range of factors, including FSH levels, results from semen analysis, and outcomes from testicular sperm aspiration. This thorough exploration aims to provide a thorough understanding of our innovative surgical technique and its application in addressing complex cases of obstructive azoospermia. Results The procedure was started on the right, the vas deferens was identified and transected. The abdominal side of the vas was intubated and a vasogram performed, there was no obstruction. There was no fluid visible from the testicular side of the vas for analysis, thus we proceeded with VE. Upon inspection of the epididymis dilated tubules were identified. After selecting a tubule for VE, two 10-0 nylon sutures were placed, and it was incised. Upon inspection of the fluid motile sperm was identified. After VE, we performed a novel epididymal occlusion stitch technique. This was completed distal to the anastomosis by placing a 7-0 prolene through the tunica of the epididymis from the medial to lateral side. This stitch was then tightened down with the goal to largely occlude the epididymis so that sperm will preferentially travel through the anastomosis. The steps were then repeated on the left. At 3-month follow up, the patient had no change in testicular size as compared with preoperative size (18cc), he had no testicular or incisional discomfort, and on semen analysis he had presence of motile sperm. After 3 months post-surgery, the patient had motile sperm seen on semen analysis. Discussion The introduction of a novel epididymal occlusion stitch demonstrates a targeted strategy to enhance the success of microscopic VE. Encouragingly, a 3-month post-surgery follow-up reveals the presence of motile sperm, reinforcing the potential efficacy of our approach. This is promising given the historical lower patency, delayed time to patency, and higher delayed failure rates that patients who require VE experience (10). In total, 40% of all azoospermia cases can be attributed to obstruction. The conventional treatments for obstructive azoospermia involve microsurgical reconstruction and surgical sperm retrieval followed by IVF. While microsurgical reconstruction has proven to be economically viable, the quest for enhanced success rates has led to the exploration of innovative techniques. Historically, the evolution of VV and VE procedures, initially performed in the early 20th century, laid the foundation for contemporary microsurgical approaches (11). Notably, the microscopic VV demonstrated significant improvements in patency rates and natural pregnancy likelihood, as evidenced by the seminal Vasovastomy Study Group study in 1991 (8). In contemporary literature, success rates particularly for VE remain unchanged for the past three decades since the original published success rates by the Vasectomy Reversal Study Group (12). VE is associated with a longer time to patency as well with patients taking 2.8 to 6.6 months to have sperm return to ejaculate as compared to 1.7 to 4.3 months for those undergoing VV. Additionally, of those patients who successfully have sperm return to the ejaculate after VE up to 50% will have delayed failure compared to 12% for those undergoing VV who are patent. Finally, of those who experience delayed failure after undergoing VE it usually occurs earlier with studies reporting as early as 6 months post-operatively (20). Given the lack of improvement and significantly worsened outcomes with VE further surgical refinement is a constant goal for surgeons performing this procedure. Conclusion: In conclusion, this video is both a demonstration and a call to action for commitment to surgical innovation. We aim to raise the bar in VE success rates, ultimately bringing tangible benefits to patients and contributing to the ongoing evolution of reproductive medicine. The novel epididymal occlusion stitch emerges as a beacon of progress, promising not only enhanced safety but also potential reductions in patency time. Surgical excellence and methodological refinement, as exemplified in this video, lay the foundation for a future where male reproductive surgery continues to break new ground. 40 1, 1 (1 2. 2 . 2) 3. (3) 4. 4 (4) vasectomy infections trauma 5. 5 (5) fertility options IVF (IVF) costeffective cost effective 6, (6 7. 7 7) However 99 (VV 65 (VE 8, 8 (8 9. 9 9) Thus 39yearold yearold 39 year old 37yearsold yearsold 37 years cycles levels aspiration right transected obstruction 100 10 0 10- placed incised 70 7- left 3month month 18cc, 18cc cc , (18cc) discomfort postsurgery, postsurgery post surgery, Encouragingly followup approach 10. (10) total viable Historically th century 11. 11 (11) Notably likelihood 199 8. (8) literature 12. 12 (12) 28 66 6. 17 1. 43 Additionally 50 patent Finally postoperatively operatively 20. 20 (20) Conclusion conclusion innovation medicine progress ground ( (3 (4 (5 (IVF (18cc (10 (11 19 (12 (20 (2

ABSTRACT: This study determined the zootechnical, metabolic, and health performance of Girolando calves born with high or low birth weight, conceived through in vitro fertilization. The study was carried out on a commercial dairy farm located in Passos, Minas Gerais, Brazil. For this, a hundred Girolando calves were divided into two groups: The Control, which consisted of calves that were born weighing ≤ 35 kg; and high birth weight (HBW) calves, which were born weighing > 35 kg. The calves were monitored for zootechnical parameters; epidemiological indices, for diseases such diarrhea and pneumonia; as well as serum concentrations of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma-glutamyl transferase, cholesterol, triglycerides, paraoxonase1, albumin, urea, and globulin. Calves from the HBW group had a higher general mortality rate, but no effects on the zootechnical performance or metabolism were observed. The results obtained allow us to conclude that Girolando calves generated by IVF and heavier at birth have a greater chance to present diseases such as omphalophlebitis, tympany, and bovine parasitic sadness, and have a higher rate of general mortality. Conversely, birth weight of the calves does not influence diarrhea or pneumonia, the zootechnical performance, or the metabolism. ABSTRACT metabolic fertilization Passos Gerais Brazil this groups Control 3 kg (HBW parameters indices pneumonia aminotransferase phosphatase gammaglutamyl gamma glutamyl transferase cholesterol triglycerides paraoxonase1 paraoxonase albumin urea globulin observed omphalophlebitis tympany sadness Conversely

RESUMO: Este estudo determinou o desempenho zootécnico, metabólico e saúde de bezerras Girolando nascidas com alto ou baixo peso, concebidas por fertilização in vitro. O estudo foi realizado em uma fazenda leiteira comercial localizada em Passos, Minas Gerais, Brasil. Cem bezerras Girolando foram divididas em dois grupos: Controle, que consistiu em bezerras que nasceram com peso ≤ 35 kg; e HBW, bezerras que nasceram com peso > 35 kg. As bezerras foram monitoradas quanto aos parâmetros zootécnicos; índices epidemiológicos para doenças como diarreia e pneumonia; bem como as concentrações séricas de aspartato aminotransferase, alanina aminotransferase, fosfatase alcalina, gama-glutamil transferase, colesterol, triglicérides, paraoxonase1, albumina, uréia e globulina. As bezerras do grupo HBW apresentaram maior taxa de mortalidade, mas nenhum efeito no desempenho zootécnico ou metabolismo foram observados. Os resultados permitem concluir que bezerras Girolando provenientes de FIV que nascem mais pesadas têm maior chance de apresentar doenças, como onfaloflebite, timpanismo e tristeza parasitária bovina, tendo uma maior taxa de mortalidade geral. Porém, o peso ao nascimento das bezerras não influenciou a ocorrência de diarreia ou broncopneumonia, o desempenho zootécnico e o perfil metabólico. RESUMO vitro Passos Gerais Brasil grupos Controle 3 kg zootécnicos pneumonia aminotransferase alcalina gamaglutamil gama glutamil transferase colesterol triglicérides paraoxonase1 paraoxonase albumina globulina observados onfaloflebite bovina geral Porém broncopneumonia

Abstract Obesity has been associated with poorer reproductive outcomes after ART treatments, affecting the quality of embryos, endometrial receptivity, and the risk of gestational complications. The decision to delay IVF/ICSI treatments to promote weight loss must be carefully considered, especially in older women. Future research should focus on weight loss interventions tailored to patients’ age and BMI, with longitudinal studies to assess the effects of these interventions. Promoting healthy lifestyles and controlled weight loss is essential, but risks and benefits must be carefully weighed to optimize reproductive treatments.

Resumo A obesidade tem sido associada a piores desfechos reprodutivos após tratamentos de PMA, afetando a qualidade embrionária, a recetividade endometrial e o risco de complicações gestacionais. A decisão de adiar tratamentos de FIV/ICSI para promover a perda de peso deve ser considerada nas mulheres mais velhas. A investigação futura deve focar-se em estudos longitudinais para avaliar os efeitos de intervenções de perda de peso adaptadas à idade e ao IMC. A promoção de estilos de vida saudáveis e a perda de peso controlada são essenciais, mas os riscos e benefícios devem ser cuidadosamente ponderados para otimizar os tratamentos reprodutivos.

ABSTRACT The aim of this study was to evaluate the effect of Insulin-Transferrin-Selenium (ITS) supplementation in embryo culture on the blastocyst rate and embryo quality. The bovine zygotes obtained by in vitro maturation (MIV) and fertilization (IVF) were transferred to Evolve medium in 4 groups (T1: Without ITS supplement; T2: ITS the first 48 h of culture (CIV1): T3: ITS after day 2 of culture (CIV2), T4: ITS during all culture time (CIV1+CIV2). All procedures were performed at 38.5 °C,5%CO2and90%relativehumidity. The cleavage and blastocyst rates were determined at 48 h and day 8 of culture. The total cell number and the production of reactive oxygen species (ROS) in the expanded blastocysts were determined by staining the nucleus with Hoechst and the oxidation of the fluorescent probe dichlorodihydrofluorescein diacetate (H2DCFDA). The T4 cultured embryos presented the highest rate of blastocysts (32.6%) and total cell number (145.6) compared to the control group without STI (26.2% and 98.6, respectively; p<0.05). Likewise, the production of ROS in T4 blastocysts was lower (p<0.05) than that obtained in the other study groups. The results show that the addition of the ITS supplement throughout the embryo culture time has beneficial effects on the percentage of blastocysts and embryo quality, assessed by the number of total cells and the production of ROS.

RESUMEN El estudio tuvo como objetivo evaluar el efecto del suplemento Insulina-Transferrina-Selenio (ITS) en el cultivo embrionario sobre la tasa de blastocistos y calidad embrionaria. Los cigotos de bovino obtenidos por maduración (MIV) y fertilización in vitro (FIV) fueron transferidos a medio Evolve en 4 grupos (T1: Sin suplemento ITS; T2: ITS las primeras 48 h de cultivo (CIV1): T3: ITS después del día 2 de cultivo (CIV2), T4: ITS todo el tiempo de cultivo (CIV1 + CIV2). Todos los procedimientos se realizaron a 38.5 °C, 5% de CO2 y 90% de humedad relativa. Las tasas de clivaje y de blastocistos se determinaron a las 48 h y día 8 de cultivo, respectivamente. El número de células totales y la producción de especies reactivas de oxígeno (EROs) en los blastocistos expandidos fueron determinados por tinción del núcleo con Hoechst y la oxidación de la sonda fluorescente diacetato diclorodihidrofluoresceína (H2DCFDA). Los embriones cultivados de T4 presentaron la mayor tasa de blastocistos (32.6%) y número de células totales (145.6) comparada al grupo control sin ITS (26.2% y 98.6, respectivamente; p<0.05). Asimismo, la producción de EROs en los blastocistos de T4 fue menor (p<0.05) que la obtenida en los demás grupos de estudio. Los resultados demuestran que la adición del suplemento ITS durante todo el tiempo de cultivo embrionario tiene efectos benéficos sobre el porcentaje de blastocistos y calidad embrionaria, valorada por el número de células totales y la producción de EROs.

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 17. The study aimed to evaluate the Fertilization Rate of four independent in vitro treatments of bovine oocytes with Sexed (SX) and Conventional (Conv) semen, capacitated (B.O and Percoll) using the Swim-Up technique. Cleavage was assessed at 48 hours post-fertilization and on the seventh day of embryonic development. The results showed a cleavage percentage of 59.4% in T1 (Conv. Percoll), 60% in T2 (Conv. B.O), 75% in T3 (SX. Percoll), and 46.6% in T4 (SX. B.O), with a statistically significant difference (p>0.05) observed specifically between T3 and T4. The embryo development rate on the seventh day was 34.4% in T1, 20% in T2, 15% in T3, and 10.51% in T4. In this case, a statistically significant difference was found in the use of conventional medium with Percoll compared to SX B.O medium (p<0.05) (T1 and T4, respectively). The results also showed similar production of morula and early blastocysts and protruded embryos among the four treatments, demonstrating the feasibility of in vitro fertilization (IVF) technique for producing bovine embryos. In conclusion, the preliminary evaluation highlights the significant influence of the sexing method and treatment on semen quality and embryonic development in animal reproduction programs. Although Sexed semen with Percoll shows potential to improve cleavage rates, conventional semen, especially in T1 treatment (Conv. Percoll), exhibits superiority in individual motility and concentration. These findings provide crucial insights for optimizing reproductive programs.

Resumen 13. El objetivo del presente estudio fue evaluar la Tasa de Fecundación in vitro de ovocitos bovinos con semen sexado (SX) y convencional (Conv.) capacitados (B.O y Percoll) con la técnica Swim-Up. Se evaluó el clivaje a las 48 horas post fecundación y al séptimo día del desarrollo embrionario. Los resultados mostraron un porcentaje de clivaje del 59,4% en el T1 (Conv. Percoll), 60% en el T2 (Conv. B.O), 75% en el T3 (SX. Percoll) y 46,6% en el T4 (SX. B.O), mostrando diferencia estadística entre T3 y T4 (p>0,05). La tasa de desarrollo del embrión al séptimo día fue 34,4% T1, 20% en T2, 15% en T3 y 10,51% en T4; en este caso, se encontró diferencia estadísticamente significativa en el uso del medio convencional más Percoll en comparación con el medio de SX B.O (p<0,05) (T1 y T4, respectivamente). Los resultados también mostraron una producción similar de mórulas y blastocitos tempranos y protruidos entre los cuatro tratamientos, y demuestran la viabilidad del desarrollo e implementación de la técnica de fertilización in vitro (FIV) para producir embriones bovinos. Concluyendo la evaluación preliminar, destaca la significativa influencia del método de sexado y del tratamiento en la calidad del semen y desarrollo embrionario en programas de reproducción animal. Aunque el semen sexado con Percoll muestra potencial para mejorar tasas de clivaje, el semen convencional, especialmente en el tratamiento T1 (Conv. Percoll), exhibe una superioridad en motilidad individual y concentración. Estos resultados informan decisiones cruciales para optimizar programas reproductivos.

Abstract The objective of the present study was to transpose sperm freezing methodology from domestic goat to the Tadjik markhor (Capra falconeri heptneri) and to address the feasibility to develop IVP and artificial insemination using such frozen semen. Semen of different adult markhor males were successfully recovered by electro-ejaculation and were then frozen using caprine methodology. Frozen semen showed good survival rates at thawing and good fertility rates were assessed in heterologous in vitro fertilization system with goat oocytes. LOPU/IVF was applied for Tadjik markhor females allowing the first successful blastocyst production in vitro. In an applied program, we also transposed successfully intrauterine AI method with frozen/thawed semen to the Tadjik markhor. Capra heptneri electroejaculation electro ejaculation oocytes LOPUIVF LOPU IVF program frozenthawed thawed

The aim of this study was to assess the effect of eugenol on the in vitro production of ovine embryos. Oocytes were aspirated and taken for in vitro maturation (IVM), where they were divided into 4 groups: the CON (control), in which the oocytes were immersed in medium without an antioxidant; in the EU10, EU20, EU40 groups, the same medium as in the COM group was used, supplemented with 10, 20, and 40 µM of eugenol, respectively. The degree of expansion of cumulus cells was evaluated. Subsequently, the oocytes underwent in vitro fertilization and in vitro culture, and at the end of this process, the rate of cleaved structures was assessed. Regarding the expansion rate of cumulus cells, the CON group exhibited higher rates compared to the groups supplemented with eugenol. Evaluating oocytes with fully expanded cumulus cells, only the EU40 group showed a lower rate when compared to the rest of the treatment groups. Whereas observing oocytes with partially expanded cumulus cells, the EU40 group showed higher expansion percentages compared to the other treatments. Concerning the presence of the first polar body in the oocytes, it was observed that there was no statistical difference between the treatment groups. Lastly, concerning the cleaved structures, the CON and EU10 groups showed the highest rates compared to the EU20 and EU40 groups. Thus, it was possible to conclude that the use of 10 µM of eugenol in the IVM of ovine oocytes proved to be the optimal concentration of this substance.

El objetivo del presente estudio fue evaluar el efecto del eugenol en la producción in vitro de embriones ovinos. Los ovocitos fueron aspirados y llevados para la maduración in vitro (MIV), donde se dividieron en 4 grupos: el CON (control), en el cual los ovocitos fueron sumergidos en medio sin antioxidantes; en los grupos EU10, EU20, EU40 se utilizó el mismo medio que en el grupo CON, pero con la adición de 10, 20 y 40 µM de eugenol, respectivamente. Se evaluó el grado de expansión de las células del cúmulo. Después de esto, los ovocitos fueron sometidos a la fecundación in vitro y al cultivo in vitro, y al final de este proceso se evaluó la tasa de estructuras clivadas. En cuanto a la tasa de expansión de las células del cúmulo, el grupo CON mostró tasas más altas en comparación con los grupos suplementados con eugenol. Al evaluar los ovocitos con células del cúmulo totalmente expandidas, solo el grupo EU40 mostró una tasa más baja en comparación con el resto de los grupos de tratamiento. Mientras que al observar los ovocitos con células del cúmulo parcialmente expandidas, el grupo EU40 mostró porcentajes de expansión más altos en comparación con los otros tratamientos. Respecto a la presencia del primer corpúsculo polar en los ovocitos, se observó que no hubo diferencia estadística entre los grupos de tratamiento. Por último, en relación con las estructuras clivadas, los grupos CON y EU10 mostraron las tasas más altas en comparación con los grupos EU20 y EU40. Por lo tanto, se pudo concluir que el uso de 10 µM de eugenol en la MIV de ovocitos ovinos resultó ser la concentración óptima de esta sustância.

Objetivando avaliar o efeito do eugenol na produção in vitro de embriões ovinos. Os oócitos, foram aspirados e levados para a maturação in vitro (MIV), onde foram divididos em 4 grupos: o CON (controle), no qual os oócitos foram imersos em meio, sem antioxidante, já nos grupos EU10, EU20, EU40 foi utilizado o mesmo meio do grupo CON, acrescido de 10, 20 e 40 µM de eugenol, respectivamente. Foi avaliado o grau de expansão das células do cumulus. Após isso, os oócitos seguiram para a fecundação in vitro e cultivo in vitro, sendo, ao final desse processo, avaliada a taxa de estruturas clivadas. Em relação à taxa de expansão das células do cumulus, o grupo CON apresentou maiores taxas, quando comparado aos grupos adicionados de eugenol. Avaliando os oócitos com células do cumulus totalmente expandidas, somente o grupo EU40, apresentou-se menor, quando comparado ao restante dos grupos de tratamento. Já observando os oócitos com células do cumulus parcialmente expandidas, o grupo EU40, apresentou maiores percentuais de expansão, quando comparado aos demais tratamentos. Sobre a presença do primeiro corpúsculo polar nos oócitos, observou-se que não houve diferença estatística entre os grupos de tratamentos. E, por último, em relação às estruturas clivadas, os grupos CON e EU10 apresentaram as maiores taxas, quando comparado aos grupos EU20 e EU40. Dessa maneira, foi possível concluir que a utilização de 10 µM de eugenol na MIV de oócitos ovinos apresentou ser a melhor concentração dessa substância.

The aim of this study was to evaluate the effect of quercetin on the in vitro maturation (IVM) of oocytes and the subsequent in vitro fertilization (IVF) of ovine embryos. For this purpose, oocytes were collected and divided into the following treatment groups: control group, containing CON medium (control); and groups Q2, Q4, and Q8, with control medium supplemented with 2, 4, and 8 µM of quercetin, respectively. After IVM, the degree of cumulus cells expansion, presence of the first polar body (PB), and the other part of the oocytes proceeded to in vitro fertilization (IVF), where oocytes were incubated with spermatozoa for a period of 20 hours. Subsequently, the presumptive zygotes underwent in vitro culture (IVC) for 48 hours, and at the end of the process, the rate of cleaved structures was evaluated. The results were expressed in percentage and compared using the Chi-square test, with a significant difference considered when P&lt;0.05. Evaluating the expansion of cumulus cells, the CON and Q2 groups showed the best expansion rates compared to the Q4 and Q8 groups. Regarding the presence of the 1st PB, it was observed that only the Q8 group exhibited lower rates compared to the control group. When evaluating the rate of cleaved structures, it was observed that the Q2 group showed a higher number compared to the Q8 group. Quercetin does not influence the maturation of ovine oocytes.

El objetivo del presente estudio fue evaluar el efecto de la quercetina en la maduración in vitro (MIV) de ovocitos y la posterior fertilización in vitro (FIV) de embriones ovinos. Para ello, se recolectaron y seleccionaron ovocitos divididos en los siguientes grupos de tratamiento: grupo control, que contenía medio CON (control); y los grupos Q2, Q4 y Q8, con medio control al que se le añadieron 2, 4 y 8 µM de quercetina, respectivamente. Tras la MIV se evaluó el grado de expansión de las células del cúmulo, la presencia del primer corpúsculo polar (CP) y la otra parte de los ovocitos se utilizó para la fecundación in vitro (FIV), donde los ovocitos fueron incubados con espermatozoides durante un período de 20 horas. Posteriormente, los presuntos cigotos pasaron a cultivo in vitro (CIV) durante 48 horas y, al final del proceso, se evaluó la tasa de estructuras clivadas. Los resultados se expresaron en porcentaje y se compararon utilizando la prueba de Chi-cuadrado, considerando una diferencia significativa cuando P&lt;0,05. Al evaluar la expansión de las células del cúmulo, los grupos CON y Q2 mostraron las mejores tasas de expansión en comparación con los grupos Q4 y Q8. En cuanto a la presencia del 1er CP, se observó que solo el grupo Q8 mostró tasas más bajas en comparación con el grupo control. Al evaluar la tasa de estructuras clivadas, se observó que el grupo Q2 mostró un número mayor en comparación con el grupo Q8. La quercetina no influye en la maduración de ovocitos ovinos.

O objetivo do presente estudo foi avaliar o efeito da quercetina na MIV de oócitos e na subsequente PIV de embriões ovinos. Para isso, foram colhidos e selecionados oócitos divididos nos seguintes grupos de tratamento: grupo controle, contendo meio CON (controle); e grupos Q2, Q4 e Q8, com meio controle, acrescido de 2, 4 e 8 µM de quercetina, respectivamente. Após a MIV foi avaliado o grau de expansão das células do cumulus, presença do primeiro corpúsculo polar (CP) e a outra parte dos oócitos seguiu para a fecundação in vitro (FIV), onde os oócitos foram incubados com os espermatozoides, por um período de 20 h. Em seguida, os presumíveis zigotos seguiram para o cultivo in vitro (CIV), por um período de 48 h e, ao final do processo, foi avaliada a taxa de estruturas clivadas. Os resultados foram expressos em porcentagem e comparados, usando o Teste do Qui-quadrado e foi considerada diferença significativa quando P&lt;0,05. Avaliando a expansão das células do cumulus, os grupos CON e Q2 apresentaram as melhores taxas de expansão, quando comparadas aos grupos Q4 e Q8. Quanto à presença do 1º CP, foi possível observar que somente o grupo Q8 apresentou taxas menores, quando comparado ao grupo controle. Já avaliando a taxa de estruturas clivadas, foi possível observar que o grupo Q2 apresentou maior número quando comparado ao grupo Q8. A quercetina não influencia na maturação de oócitos ovinos.

The objective of this study was to evaluate the effect of α-lipoic acid on in vitro maturation and of l-arginine on in vitro fertilization of oocytes for in vitro production of sheep embryos. Thus, the ovaries were collected at a local slaughterhouse and the oocytes collected by the method of follicular aspiration with a vacuum pump were divided equally into five maturation groups: CON, without addition of antioxidant; CIS group, with cysteamine as an antioxidant source; and the groups ALA5, ALA10 and ALA20, containing concentrations of 5.0, 10.0 and 20.0 µM of ALA. After 24h of maturation, the oocytes were evaluated concerning occurrence and degrees of expansion of the cumulus cells. The oocytes of the best IVM group were destinated for in vitro fertilization (IVF). The selected and capacitated sperm were included in five groups: CON, without addition of capacitator agent; HEP group, with heparin as a capacitator source; and ARG5, ARG10 and ARG20 groups, where IVF occurred in the same HEP medium, replacing heparin with 5 mM, 10 mM and 20 mM L-arginine, respectively. The results were expressed as a percentage and the variables of cumulus cell expansion and number of cleaved structures were compared using the chi-square test in Epi Info software (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Results were considered significant when P &lt; 0.05. Regarding the expansion of cumulus cells all groups showed 100% expansion, on the total expansion category of COCs it was observed that ALA5 and ALA20 groups did not present the same proportion of COCs with total expansion as CIS group. After in vitro fertilization (IVF), there were no statistical differences in relation to the cleaved structures between CON, HEP, ARG5 and ARG10 groups. However, it was also observed that the use of 20.0 mM of L-arginine reduced the percentage of cleaved structures when compared to the ARG10 group (P&lt;0.05).

El objetivo de este trabajo fue evaluar el efecto del ácido α-lipoico en la maduración in vitro y la l-arginina en la fecundación in vitro de ovocitos para la producción in vitro de embriones ovinos. Así, se recolectaron ovocitos de ovarios obtenidos de mataderos mediante el método de aspiración folicular con bomba de vacío, los cuales se dividieron en cinco grupos de maduración: CON, sin adición de antioxidante, grupo CIS, con cisteamina como fuente antioxidante y los grupos ALA5, ALA10 y ALA20, que contienen concentraciones de 5, 10 y 20 µM de ácido α-lipoico. Después de 24 h de maduración, se evaluó la aparición y el grado de expansión de las células del cúmulo en los ovocitos. Los ovocitos del mejor grupo MIV fueron destinados a fecundación in vitro (FIV), siendo por tanto incluidos en gotas con medio de fecundación in vitro, junto con los espermatozoides seleccionados y capacitados, resultando en los siguientes grupos: CON, sin adición de agente capacitador; grupo HEP, con heparina como fuente de condensadores; y los grupos ARG5, ARG10 y ARG20, donde se produjo la FIV en el mismo medio HEP, reemplazando la heparina por L-arginina 5, 10 y 20 mM, respectivamente. Los resultados se expresaron en porcentajes y las variables expansión celular del cúmulo y número de estructuras escindidas se compararon mediante la prueba chicuadrado del software Epi Info (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Los resultados se consideraron significativos cuando P lt; 0,05. En cuanto a la expansión de las células del cúmulo, todos los grupos mostraron 100% de expansión. Evaluando los grados de expansión de las células del cúmulo, en la categoría de expansión total, se observó que los grupos ALA5 y ALA20 presentaron la misma proporción de CCO con expansión total que el grupo CIS (Plt;0.05), sin embargo se observa que hay una tendencia a la baja de las estructuras con plena expansión cuando se utilizan antioxidantes. En cuanto a las estructuras escindidas, fue posible identificar que no hubo diferencias estadísticas en relación a las estructuras escindidas, entre los grupos CON, HEP, ARG5 y ARG10. Sin embargo, también se observó que el uso de Larginina 20 mM redujo el porcentaje de estructuras escindidas en comparación con el grupo ARG10 (Plt;0.05).

O objetivo deste trabalho foi avaliar o efeito do ácido α-lipóico na maturação in vitro e da l-arginina na fertilização in vitro de oócitos para produção in vitro de embriões ovinos. Desta forma, foram realizadas colheitas de oócitos oriundos de ovários obtidos em abatedouro através do método de aspiração folicular com bomba de vácuo, que foram divididos em cinco grupos de maturação: CON, sem adição de antioxidante, grupo CIS, com cisteamina como fonte antioxidante e os grupos ALA5, ALA10 e ALA20, contendo as concentrações de 5, 10, e 20 µM de ácido α-lipóico. Após 24h de maturação, os oócitos foram avaliados quanto à ocorrência e grau de expansão das células do cumulus. Os oócitos do melhor grupo MIV foram destinados à fertilização in vitro (FIV), sendo, portanto, incluídos em gotas com meio de fertilização in vitro, juntamente com os espermatozoides selecionados e capacitados, resultando nos seguintes grupos: CON, sem adição de capacitador agente; grupo HEP, com heparina como fonte de capacitor; e grupos ARG5, ARG10 e ARG20, onde a FIV ocorreu no mesmo meio HEP, substituindo a heparina por L-arginina 5, 10 e 20 mM, respectivamente. Os resultados foram expressos em porcentagem e as variáveis de expansão das células do cumulus e número de estruturas clivadas foram comparadas por meio do teste qui-quadrado do software Epi Info (Epi Info 7.2.5, Atlanta, GA, EUA, 2021). Os resultados foram considerados significativos quando P lt; 0,05. Em relação à expansão das células do cumulus, todos os grupos apresentaram 100% de expansão. Avaliando os graus de expansão das células do cumulus, na categoria de expansão total observou-se que os grupos ALA5 e ALA20 apresentaram a mesma proporção de CCO com expansão total que o grupo CIS (Plt;0,05), porém se observa que há uma tendência de queda de estruturas com expansão total quando os antioxidantes foram utilizados. Em relação às estruturas clivadas, foi possível identificar que não houve diferenças estatísticas em relação às estruturas clivadas, entre os grupos CON, HEP, ARG5 e ARG10. Contudo, também foi observado que o uso de 20 mM de L-arginina reduziu o percentual de estruturas clivadas quando comparado ao grupo ARG10 (Plt;0,05).

The objective of this study was to evaluate the effect of including cilostazol in the in vitro maturation medium of oocytes on the in vitro production of sheep embryos. Oocytes were collected from ovaries obtained from a slaughterhouse by follicular aspiration with a vacuum pump. The oocytes were divided into four maturation groups: the CON group, where the cumulus-oocyte complexes were immersed in TCM-199 supplemented with 500 IU of penicillin, 0.5 mg of streptomycin, 1.25 µg of amphotericin, 0.2 mM of sodium pyruvate, 10% (v/v) fetal bovine serum (FBS), 10 ng/mL of epidermal growth factor (EGF), 10 µg/mL of FSH, 10 µg/mL of LH, 10 µg/mL of estradiol, and 100 µM of cysteamine; and in the CILO0.3, CILO1, and CILO10 groups, the oocytes were matured in the CON group medium without the addition of cysteamine and supplemented with concentrations of 0.3, 1, and 10 µM of cilostazol, respectively. After 24 hours, the oocytes were evaluated for the presence or absence of cumulus cells and the degree of expansion and then subjected to in vitro fertilization (IVF) with sperm in FIV medium. After IVF, the presumptive zygotes were cultured in vitro. Cleavage was evaluated on day 2, with day 0 being the start of IVF. Results were expressed as a percentage, and variables such as cumulus cell expansion and the number of cleaved structures were compared using the chi-square test in the Epi Info software (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Results were considered significant when P lt; 0.05. All groups showed 100% cumulus cell expansion, and there were no significant differences in cumulus cell expansion degree between the cilostazol- and cysteamine-supplemented groups (P gt; 0.05), as well as no significant differences in cleavage rates between the cilostazol- and cysteamine-supplemented groups (P gt; 0.05).

El objetivo de este estudio fue evaluar el efecto de la inclusión de cilostazol en el medio de maduración in vitro de ovocitos sobre la producción in vitro de embriones ovinos. Para ello, se realizaron recolecciones de ovocitos provenientes de ovarios obtenidos en un matadero mediante el método de aspiración folicular con bomba de vacío. Los ovocitos se dividieron en cuatro grupos de maduración: grupo CON, donde los complejos cúmulus ovocitos se sumergieron en TCM-199, suplementado con 500 UI de penicilina, 0,5 mg de estreptomicina, 1,25 µg de anfotericina, 0,2 mM de piruvato de sodio, 10% (v/v) de suero fetal bovino (SFB), 10 ng/mL de factor de crecimiento epidérmico (EGF), 10 µg/m de FSH, 10 µg/mL de LH, 10 µg/mL de estradiol y 100 µM de cisteamina; y en los grupos CILO0,3; CILO1 y CILO10, los ovocitos se maduraron en el medio del grupo CON, pero sin la adición de cisteamina y suplementado con las concentraciones de 0,3; 1 y 10 µM, respectivamente. Después de 24 horas, los ovocitos se evaluaron en cuanto a la presencia o no de células del cúmulus y en cuanto al grado de expansión y se destinaron a la fecundación in vitro, en medio FIV, junto con espermatozoides. Después de la FIV, los presuntos cigotos siguieron para el cultivo in vitro. Se evaluaron las clivajes en el día 2, siendo el día 0 el día del inicio del CIV. Los resultados se expresaron en porcentaje y las variables de expansión de las células del cúmulus y número de estructuras clivadas se compararon mediante la prueba del chi-cuadrado del software Epi Info (Epi Info 7.2.5, Atlanta, GA, EE. UU., 2021). Los resultados se consideraron significativos cuando P lt; 0,05. En relación a la expansión de las células del cúmulus, todos los grupos presentaron el 100% de expansión. No hubo diferencias significativas en cuanto al grado de expansión de las células del cúmulus entre los grupos suplementados con cilostazol y cisteamina (P gt; 0,05), así como no hubo diferencias significativas entre las tasas de clivaje entre los grupos suplementados con cilostazol y cisteamina (P gt; 0,05).

O objetivo deste estudo foi avaliar o efeito da inclusão de cilostazol no meio de maturação in vitro de oócitos sobre produção in vitro de embriões ovinos. Para isso, foram realizadas colheitas de oócitos oriundos de ovários obtidos em abatedouro por meio do método de aspiração folicular com bomba de vácuo. Os oócitos foram divididos em quatro grupos de maturação: grupo CON, onde os complexos cumulus oócitos foram imersos em TCM-199, suplementado com 500 UI de penicilina, 0,5 mg de estreptomicina, 1,25 µg de anfotericina, 0,2 mM de piruvato de sódio, 10% (v/v) de soro fetal bovino (SFB), 10 ng/mL de fator de crescimento epidérmico (EGF), 10 µg/m de FSH, 10 µg/mL de LH, 10 µg/mL de estradiol e 100 µM de cisteamina; e nos grupos CILO0,3; CILO1 e CILO10, os oócitos foram maturados no meio do grupo CON, mas sem a adição de cisteamina e suplementado com as concentrações de de 0,3; 1 e 10 µM , respectivamente. Após 24h, os oócitos foram avaliados quanto a presença ou não de células do cumulus e quanto ao grau de expansão e destinados à fecundação in vitro, em meio FIV, juntamente com espermatozoides. Após a FIV, os presumíveis zigotos seguiram para o cultivo in vitro. Foram avaliadas clivagens no dia 2, sendo dia 0 o dia do início do CIV. Os resultados foram expressos em porcentagem e as variáveis de expansão das células do cumulus e número de estruturas clivadas foram comparadas por meio do teste qui-quadrado do software Epi Info (Epi Info 7.2.5, Atlanta, GA, EUA, 2021). Os resultados foram considerados significativos quando P lt; 0,05. Em relação à expansão das células do cumulus, todos os grupos apresentaram 100% de expansão. Não houve diferenças significativas quanto ao grau de expansão das células do cumulus entre os grupos suplementados com cilostazol e cisteamina (P gt; 0,05), assim como não houve diferenças significativas entre as taxas de clivagem entre os grupos suplementados com cilostazol e cisteamina (P gt; 0,05).

Abstract Objective To understand the impact of the coronavirus disease 2019 pandemic on in vitro fertilization (IVF) clinical pregnancy rates and analyze factors that may have influenced their outcome. Methods This was a retrospective observational study conducted at a tertiary-care Brazilian fertility center. All fresh IVF and embryo warming cycles performed from March 11 to December 31, 2018–2021 were analyzed, and their data were used to calculate fertilization, embryo cleavage, cycle cancellation, embryo transfer (ET), and clinical pregnancy rates. Statistical tests were used to evaluate the alterations found. Logistic regression models were used to explore the association of the categorical variables with the observed clinical pregnancy rates. Data from 2018 and 2019 (prepandemic) and 2020 and 2021 (pandemic) were grouped. Results A total of 756 cycles were analyzed (n = 360 prepandemic and n = 396 pandemic). The age group of the patients, fertilization rates, and cleavage rates did not have significant differences (p > 0.05). There was a reduction in the percentage of fresh IVF and an increase in embryo warming cycles (p = 0.005) during the pandemic. There was also an increase in fresh cycle cancellations (p < 0.001) and a reduction in ET rates (p < 0.001). The pandemic had a negative impact on clinical pregnancy rates (p < 0.001) especially due to the increase in fresh cycle cancellations (p < 0.001). Conclusion Embryo warming cycles with subsequent frozen-thawed ET were presented as a viable alternative to continue assisted reproductive treatments against pandemic restrictions on fresh cycles, ensuring clinical pregnancy, albeit at a lower rate than that of the prepandemic period. 201 (IVF outcome tertiarycare tertiary care center 1 31 20182021 2018–202 cancellation ET, , (ET) found (prepandemic 202 (pandemic grouped 75 36 39 . pandemic) patients p 0.05. 005 0.05 0 05 0.05) 0.005 0005 0.001 0001 001 0.001. frozenthawed frozen thawed period 20 3 2018202 2018–20 (ET 7 00 0.0 0.00 000 2 201820 2018–2 0. 20182 2018–

Resumo Objetivo Compreender os impactos da pandemia de COVID-19 nas taxas de gravidez clínica em fertilização in vitro (FIV) e analisar fatores que possam ter influenciado seu resultado. Métodos Foi realizado um estudo observacional retrospectivo em um centro brasileiro de reprodução assistida. Todos os ciclos de FIV com embriões frescos e descongelados realizados entre 11 de março e 31 de dezembro, 2018-2021 foram analisados, e seus dados utilizados para cálculo das taxas de fertilização, clivagem embrionária, cancelamento de ciclos, transferência de embriões (TE) e gravidez clínica. Testes estatísticos avaliaram significância das alterações encontradas e modelos de regressão logística exploraram associação das variáveis categóricas estudadas com as taxas de gravidez clínica observadas. Os dados de 2018 e 2019 (pré-pandemia) e 2020 e 2021 (pandemia) foram agrupados. Resultados Foram analisados um total de 756 ciclos (n = 360 na pré-pandemia e n = 396 na pandemia). A faixa etária das pacientes e as taxas de fertilização e de clivagem não tiveram alterações significativas (p > 0,05). Na pandemia, houve redução da porcentagem de ciclos de FIV com embriões frescos e aumento dos com descongelamento (p = 0,005). Também foi notado aumento das taxas de cancelamentos de ciclos com embriões frescos (p < 0,001) e redução do número de TEs (p < 0,001). A pandemia exerceu impacto negativo na taxa de gravidez clínica (p < 0,001), especialmente devido ao aumento de cancelamentos dos ciclos a fresco (p < 0,001). Conclusão Frente às limitações pandêmicas impostas aos ciclos com embriões frescos, os ciclos de descongelamento de embriões se apresentaram como alternativa viável à continuidade dos ciclos de FIV, garantindo gravidez clínica ainda que em taxas inferiores às do período pré-pandêmico. COVID19 COVID 19 COVID-1 (FIV resultado assistida 1 3 dezembro 20182021 2018-202 embrionária TE (TE observadas 201 prépandemia pré (pré-pandemia 202 (pandemia agrupados 75 36 39 pandemia. . pandemia) p 0,05. 005 0,05 0 05 0,05) 0,005. 0005 0,005 0,005) 0,001 0001 001 0,001. 0,001, , prépandêmico. prépandêmico pandêmico. pandêmico pré-pandêmico COVID1 COVID- 2018202 2018-20 20 7 00 0,0 000 0,00 201820 2018-2 2 0, 20182 2018-

ABSTRACT Objective: To evaluate the effect of metabolic syndrome (MetS) diagnosis on oocyte quality and pregnancy outcomes in infertile women with polycystic ovary syndrome (PCOS) who undergoing antagonist-controlled ovarian stimulation (COS) and in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Subject and methods: This prospective cohort study was conducted from November 2019 to November 2020 across two university-affiliated infertility centers in Iran. The PCOS diagnosis was defined according to the Rotterdam criteria. The patients prior to IVF/ICSI cycles were evaluated for MetS diagnosis. MetS was detected according to the National Cholesterol Education Program/Adult Treatment Panel III with the presence of at least three or more of the specific clinical criteria. The cycle outcomes were compared between MetS and non-MetS groups. Results: Overall, 68 eligible infertile PCOS patients with MetS diagnosis and 126 without MetS participated. The MetS diagnosis was associated with the increased requirement of gonadotropins and the COS duration significantly (P = 0.001). Although the total numbers of retrieved and MII oocytes, obtained and top-quality embryos as well as clinical pregnancy and live birth rates in the MetS group were lower than those of in the non-MetS group, the differences were not statistically significant (P > 0.05). In follow-up of the obstetrics complications, the rate of preeclampsia was significantly higher in patients with MetS (P = 0.02). Conclusion: MetS diagnosis in PCOS patients was associated with non-significant poor COS and pregnancy outcome. Further studies with larger sample sizes are recommended to clarify the risk of MetS in patients undergoing ART cycles. Objective (MetS (PCOS antagonistcontrolled antagonist controlled (COS fertilizationintracytoplasmic fertilization intracytoplasmic IVFICSI IVF ICSI (IVF/ICSI methods 201 202 universityaffiliated university affiliated Iran criteria ProgramAdult Program Adult nonMetS non groups Results Overall 6 12 participated P 0.001. 0001 0.001 . 0 001 0.001) oocytes topquality top 0.05. 005 0.05 05 0.05) followup follow up complications 0.02. 002 0.02 02 0.02) Conclusion nonsignificant outcome 20 1 000 0.00 00 0.0 2 0.

In biliary atresia (BA), efforts to prevent premature liver transplantation (LT) are aimed at early diagnosis, timing of Kasai-portoenterostomy (KPE), and centralization of care. This report presents the clinical picture, treatment strategies, and outcomes of BA patients with no previous treatment. A retrospective cohort study (Jan/2001 to Jan/2021) was conducted to evaluate the outcome of patients with BA referred to a single team. Study groups were: 1) Kasai-only group (K-only) n=9), 2) LT-only group (n=7), and 3) Kasai+LT group (K+LT) (n=23). Survival with native liver and overall survival were 22.9 and 94.8%, respectively, at 120 months of follow-up. There was no difference in age at KPE in the K-only group (46.8±21.8 days) vs K+LT (52.1±22 days), P=0.4. Ten (25.6%) patients were babies conceived through in vitro fertilization (IVF). Four IVF patients (40%) presented associated congenital heart disease vs 5 patients (17%) in the remaining group (P=0.14). Two of the IVF patients were premature (<37 weeks). Median maternal age at birth was 35 years (33 to 41 years). Excellent patient survival is expected for patients with BA with the available treatment strategies. IVF+BA was an unexpected prevalent association in this cohort, and further studies are required to better understand these findings. BA, , (BA) LT (LT diagnosis Kasaiportoenterostomy Kasai portoenterostomy KPE, (KPE) care picture strategies Jan/2001 Jan2001 Jan 2001 (Jan/200 Jan/2021 Jan2021 2021 team 1 Kasaionly only Konly K (K-only n=9, n9 n n=9 9 n=9) 2 LTonly n=7, n7 n=7 7 (n=7) 3 KasaiLT KLT (K+LT n=23. n23 n=23 . 23 (n=23) 229 22 22. 948 94 8 94.8% respectively 12 followup. followup follow up. up follow-up 46.8±21.8 468218 46 21 (46.8±21. days 52.1±22 52122 52 (52.1±2 days, P04 P 0 4 P=0.4 25.6% 256 25 6 (25.6% IVF. (IVF) 40% 40 (40% 17% 17 (17% P=0.14. P014 P=0.14 14 (P=0.14) <37 37 (<3 weeks. weeks weeks) 33 (3 years. years) IVFBA findings (BA (KPE Jan/200 Jan200 200 (Jan/20 Jan/202 Jan202 202 n= (n=7 n2 n=2 (n=23 94.8 46.8±21. 46821 (46.8±21 52.1±2 5212 (52.1± P0 P=0. 25.6 (25.6 (IVF (40 (17 P01 P=0.1 (P=0.14 <3 (< ( Jan/20 Jan20 20 (Jan/2 (n= (n=2 94. 46.8±21 4682 (46.8±2 52.1± 521 (52.1 P=0 25. (25. (4 (1 (P=0.1 < Jan/2 Jan2 (Jan/ (n 46.8±2 468 (46.8± 52.1 (52. P= (25 (P=0. Jan/ (Jan 46.8± (46.8 52. (52 (2 (P=0 46.8 (46. (5 (P= 46. (46 (P

Abstract Despite relatively high maturation rate of in vitro matured oocytes in the dromedary camel, however, blastocyst production is very low after in vitro fertilization (IVF). Herein, the influences of oocyte collection method (follicular aspiration vs slicing; Experiment I), the addition of Insulin-like growth factor I (IGF-I) to the maturation medium (Experiment II) on in vitro maturation (IVM) of oocyte were investigated. Although the nuclear maturation did not differ regardless of collecting method, follicular aspiration led to lower degeneration rates than those in controls (P < 0.05). The percentages of oocytes at MII were greater in the presence of IGF-1 than in its absence (71.9% vs 48.4%, respectively, P<0.05). Additionally, the percentages of degenerated oocytes were higher in the control group compared to oocytes cultured in the presence of IGF-I (23.6% vs 10.4%, respectively, P<0.05). IGF-I treatment improved the quality of MII matured oocytes as evidenced by the decrease of cathepsin B (CTSB) activity, a marker of poor quality oocytes, when compared to control ones (P < 0.05). In conclusion, follicular aspiration decreased the degeneration rate; however, it had no effect on completion of maturation. IGF-I enhanced the IVM of oocyte and decreased degeneration rate. camel however IVF. IVF . (IVF) Herein slicing I, , I) Insulinlike Insulin like IGFI IGF (IGF-I II (IVM investigated P 0.05. 005 0.05 0 05 0.05) IGF1 1 IGF- 71.9% 719 71 9 (71.9 484 48 4 48.4% respectively P<0.05. P005 P<0.05 P<0.05) Additionally 23.6% 236 23 6 (23.6 104 10 10.4% CTSB (CTSB activity conclusion (IVF 00 0.0 71.9 7 (71. 48.4 P00 P<0.0 23.6 2 (23. 10.4 0. 71. (71 48. P0 P<0. 23. (23 10. (7 P<0 (2 ( P<