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In reproductive technologies, uncovering the molecular aspects of oocyte and embryo competence under different conditions is crucial for refining protocols and enhancing efficiency. RNA-seq generates high-throughput data and provides transcriptomes that can undergo additional computational analyses. This study presented the transcriptomic profiles of in vitro matured oocytes and blastocysts produced in vitro from buffalo crossbred (Bubalus bubalis), coupled with gene co-expression and module preservation analysis. Cumulus Oophorus Complexes, obtained from slaughterhouse-derived ovaries, were subjected to in vitro maturation to yield metaphase II oocytes (616) or followed in vitro fertilization and culture to yield blastocysts for sequencing (526). Oocyte maturation (72%, ±3.34 sd) and embryo development (21.3%, ±4.18 sd) rates were obtained from three in vitro embryo production routines following standard protocols. Sequencing of 410 metaphase II oocytes and 70 hatched blastocysts (grade 1 and 2) identified a total of 13,976 genes, with 62% being ubiquitously expressed (8,649). Among them, the differentially expressed genes (4,153) and the strongly variable genes with the higher expression (fold-change above 11) were highlighted in oocytes (BMP15, UCHL1, WEE1, NLRPs, KPNA7, ZP2, and ZP4) and blastocysts (APOA1, KRT18, ANXA2, S100A14, SLC34A2, PRSS8 and ANXA2) as representative indicators of molecular quality. Additionally, genes exclusively found in oocytes (224) and blastocysts (2,200) with specific biological functions were identified. Gene co-expression network and module preservation analysis revealed strong preservation of functional modules related to exosome components, steroid metabolism, cell proliferation, and morphogenesis. However, cell cycle and amino acid transport modules exhibited weak preservation, which may reflect differences in embryo development kinetics and the activation of cell signaling pathways between buffalo and bovine. This comprehensive transcriptomic profile serves as a valuable resource for assessing the molecular quality of buffalo oocytes and embryos in future in vitro embryo production assays.

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In this longitudinal study, the anti-Müllerian hormone (AMH) levels in blood were determined in 32 Murrah buffalo females at 8, 10, 12, 16 and 19 months of age when females were synchronized and the antral follicular population (AFP) was estimated. Correlations of AFP to the AMH level at 19 months of age and retrospectively to younger ages were investigated. Then females were split into high and low AFP, and their AMH levels were compared for all ages and tested as predictors of AFP categories. The highest AMH level (p < .05) was detected at 8 months, reducing but not differing (p > .05) at 10, 12 and 16 months then reducing again (p < .05) at 19 months of age. The mean AFP was 17.6 ± 6.3 follicles, and it was positively correlated with AMH in all ages tested. High AFP females had approximately two times more antral follicles than low AFP (p < .05) and their AMH levels were higher (p < .01) than in low AFP ones in all ages. Only at 8 months, AMH levels can be used to precociously detect high AFP heifers (a cut-off point of 464.7 pg/mL; p < .05), while low AFP heifers could be detected by AMH measurements at 8, 10, 12 and 16 months of age (p < .05). We conclude that AMH of buffalo calves correlates with AFP of heifers later in life and depending on the age, its levels could be used to identify future females with low or high AFP.

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Testicular ultrasound enables the evaluation of changes in the testicular parenchyma. This study aimed to report the occurrence of hypoechogenic testicular alterations and their relationship with semen quality in five breeding buffaloes. Two buffaloes presented with hyperechoic points characteristic of fibrosis and anechoic density content between the parietal and visceral tunica. The two bulls without ultrasonographic changes showed higher average trajectory speed, linear velocity, curvilinear velocity, amplitude of lateral displacement of the spermatic head, total motility, progressive motility, fast speed, and acrosomal membrane values within the normal range. The number of spermatozoa with major and total defects was higher in the group of animals without alterations. The three buffaloes that presented with testicular alterations produced semen within established freezing standards.

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A puberdade corresponde ao momento da primeira ovulação, que culmina com o desenvolvimento final dos órgãos e estruturas envolvidas no controle neuro-endócrino reprodutivo. É um fenômeno multifatorial com grande impacto na vida produtiva das futuras matrizes e em sua eficiência econômica, ocorrendo muitas interações entre os fatores internos e externos. A idade da puberdade está relacionada ao peso corporal, que possui grandes efeitos da genética do animal e sua interação com o ambiente, especialmente a nutrição e interação social com machos. Algumas espécies, como os bubalinos, sofrem efeito de outros elementos ambientais, como a luminosidade e o fotoperíodo. A puberdade novilhas búfalas ou bovinas pode ser adiantada por meio do emprego de biotécnicas: indução da puberdade ou inseminação artificial em tempo fixo (IATF). Em ambas, os protocolos baseiam-se, principalmente, em aplicação de progestágeno associado com fonte de estradiol; mas a indução também pode ser realizada utilizando outros hormônios, como hormônio liberador de gonadotrofinas, prostaglandinas e gonadotrofina coriônica equina. No emprego da IATF, novilhas pré-púberes podem ser induzidas e inseminadas com outro protocolo de sincronização ou podem ser inseminadas com o mesmo protocolo de IATF, podendo aumentar as taxas de prenhez e diminuir os custos. Adiantando a puberdade, ocorre incremento na produção, melhorando a eficiência econômica da atividade pecuária.(AU)

Puberty corresponds to the moment of the first ovulation, which culminates in the final development of the organs and structures involved in the neuro-endocrine control of reproduction. It is a multifactorial phenomenon with a great impact on the productive life of future matrices and their economic efficiency, with many interactions between internal and external factors. The age of puberty is related to body weight, which has great effects on the animal's genetics and its interaction with the environment, especially nutrition and social interaction with males. Some species, such as buffalo, are affected by other environmental elements, such as luminosity and photoperiod. Puberty in buffaloes and bovine heifers can be advanced through the use of biotechniques: induction of puberty or timed artificial insemination (TAI). In both, the protocols are mainly based on the application of progestogen associated with a source of estradiol; but induction can also be performed using other hormones, such as gonadotropin-releasing hormone, prostaglandins and equine chorionic gonadotropin. In the employment of the TAI, prepubertal heifers can be induced and inseminated with another synchronization protocol or they can be inseminated with the same TAI protocol, which can increase pregnancy rates and decrease costs. Advancing puberty, there is an increase in production, improving the economic efficiency of livestock activity.(AU)

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Sex selection through sperm sorting offers advantages in regards selection pressure in high-producing livestock. However, the sex-sorting process results in sperm membrane and DNA damage that ultimately decrease fertility. We hypothesized that given the role of protamines in DNA packaging, protamine deficiency could account, at least partially, for the DNA damage observed following sperm sex sorting. To test this, we compared protamine status between unsexed and sexed spermatozoa from two bulls using the fluorochrome chromomycin A3 (CMA3) and flow cytometry. Then, we assessed embryo development following in vitro fertilization (IVF) using the same sperm treatments. Overall, sperm protamination was not different between sexed and unsexed semen. However, one of the two bulls displayed higher rates of protamine deficiency for both unsexed and sexed semen (P < 0.05). Moreover, unsexed semen from this bull yielded lower blastocyst (P < 0.05) and blastocyst hatching rates than unsexed sperm from the other bull. CMA3-positive staining was negatively correlated with cleavage (R2 85.1, P = 0.003) and blastocyst hatching (R2 87.6, P = 0.006) rates in unsexed semen. In conclusion, while the sex-sorting process had no effect on sperm protamine content, we observed a bull effect for sperm protamination, which correlated to embryo development rates following IVF.

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Ruminant energy supplementation with vegetable oils or fats has been standing out worldwide and oil palm processing has been receiving growing interest. This study assessed the effect of supplementation with saturated and unsaturated fatty acids from the palm oil industry on the lipid profile of seminal plasma and of the sperm membrane, as well as on the morphological and functional characteristics of raw and cryopreserved buffalo semen. Twelve purebred Murrah bulls (Bubalus bubalis) were assigned to the experimental groups and fed diets for 120 days with no added lipids (CONT, four bulls), or with an extra amount of 3% lipids from crude palm oil (PALM, four bulls), or from palm oil deodorizer distillate (PODD, four bulls). Semen was collected and cryopreserved every 15 days. The lipid composition of membranes and semen quality were determined after collections. Lipid supplementation did not impact feed intake (P>0.05). Diet enrichment with PALM increased the linoleic acid (C18:2,ω6) in seminal plasma. Lipid supplementation did not increase the polyunsaturated fatty acids in the sperm membrane composition, but significantly increased the lignoceric acid (C24:0). Cryopreserved semen of the supplemented bulls presented higher progressive motility (60.2 vs. 67.9 vs. 65.2%; P<0.05) and sperm viability detected by eosin-nigrosin staining (61.1 vs. 69.4 vs. 67.8%; P<0.05). Palm oil reduced major sperm defects in both raw (12.2 vs. 9.3 vs. 13.2%; P<0.0001) and cryopreserved semen (12.4 vs. 9.4 vs. 11.2%; P<0.0001). The lipids added to the diet did not impact the population of spermatozoa with intact plasma and acrosomal membranes (PI-/PSA-), but significantly increased the percentage of spermatozoa with high mitochondrial potential (25.6 vs. 31.5 vs. 32.0%; P=0.008). The results suggest that lipid supplementation based on crude palm oil or palm oil deodorizer distillate can be safely used to feed buffalo bulls and may increase sperm attributes related to male fertility.

RESUMO

Ruminant energy supplementation with vegetable oils or fats has been standing out worldwide and oil palm processing has been receiving growing interest. This study assessed the effect of supplementation with saturated and unsaturated fatty acids from the palm oil industry on the lipid profile of seminal plasma and of the sperm membrane, as well as on the morphological and functional characteristics of raw and cryopreserved buffalo semen. Twelve purebred Murrah bulls (Bubalus bubalis) were assigned to the experimental groups and fed diets for 120 days with no added lipids (CONT, four bulls), or with an extra amount of 3% lipids from crude palm oil (PALM, four bulls), or from palm oil deodorizer distillate (PODD, four bulls). Semen was collected and cryopreserved every 15 days. The lipid composition of membranes and semen quality were determined after collections. Lipid supplementation did not impact feed intake (P>0.05). Diet enrichment with PALM increased the linoleic acid (C18:2,6) in seminal plasma. Lipid supplementation did not increase the polyunsaturated fatty acids in the sperm membrane composition, but significantly increased the lignoceric acid (C24:0). Cryopreserved semen of the supplemented bulls presented higher progressive motility (60.2 vs. 67.9 vs. 65.2%; P 0.05) and sperm viability detected by eosin-nigrosin staining (61.1 vs. 69.4 vs. 67.8%; P 0.05). Palm oil reduced major sperm defects in both raw (12.2 vs. 9.3 vs. 13.2%; P 0.0001) and cryopreserved semen (12.4 vs. 9.4 vs. 11.2%; P 0.0001). The lipids added to the diet did not impact the population of spermatozoa with intact plasma and acrosomal membranes (PI-/PSA-), but significantly increased the percentage of spermatozoa with high mitochondrial potential (25.6 vs. 31.5 vs. 32.0%; P=0.008). The results suggest that lipid supplementation based on crude palm oil or palm oil deodorizer distillate can be safely used to feed buffalo bulls and may increase sperm attributes related to male fertility.(AU)

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RESUMO

Ruminant energy supplementation with vegetable oils or fats has been standing out worldwide and oil palm processing has been receiving growing interest. This study assessed the effect of supplementation with saturated and unsaturated fatty acids from the palm oil industry on the lipid profile of seminal plasma and of the sperm membrane, as well as on the morphological and functional characteristics of raw and cryopreserved buffalo semen. Twelve purebred Murrah bulls (Bubalus bubalis) were assigned to the experimental groups and fed diets for 120 days with no added lipids (CONT, four bulls), or with an extra amount of 3% lipids from crude palm oil (PALM, four bulls), or from palm oil deodorizer distillate (PODD, four bulls). Semen was collected and cryopreserved every 15 days. The lipid composition of membranes and semen quality were determined after collections. Lipid supplementation did not impact feed intake (P>0.05). Diet enrichment with PALM increased the linoleic acid (C18:2,6) in seminal plasma. Lipid supplementation did not increase the polyunsaturated fatty acids in the sperm membrane composition, but significantly increased the lignoceric acid (C24:0). Cryopreserved semen of the supplemented bulls presented higher progressive motility (60.2 vs. 67.9 vs. 65.2%; P 0.05) and sperm viability detected by eosin-nigrosin staining (61.1 vs. 69.4 vs. 67.8%; P 0.05). Palm oil reduced major sperm defects in both raw (12.2 vs. 9.3 vs. 13.2%; P 0.0001) and cryopreserved semen (12.4 vs. 9.4 vs. 11.2%; P 0.0001). The lipids added to the diet did not impact the population of spermatozoa with intact plasma and acrosomal membranes (PI-/PSA-), but significantly increased the percentage of spermatozoa with high mitochondrial potential (25.6 vs. 31.5 vs. 32.0%; P=0.008). The results suggest that lipid supplementation based on crude palm oil or palm oil deodorizer distillate can be safely used to feed buffalo bulls and may increase sperm attributes related to male fertility.

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In Vitro Embryo Production (IVP) is widely used to improve the reproductive efficiency of livestock animals, however increasing the embryo development rates and pregnancy outcomes is still a challenge for some species. Thus, the lack of biological knowledge hinders developing specie-specific IVP protocols. Therefore, the contributions of RNA-seq to generate relevant biological knowledge and improve the efficiency of IVP in livestock animals are reviewed herein.

RESUMO

The present study evaluated Brangus cows treated with single doses of follicle stimulating hormone (FSH) subjected to follicular aspiration after 24 h to assess oocyte recovery, in vitro fertilization and pregnancy rate. Follicles exceeding 3 millimeters in diameter were aspirated, 200 mg of FSH was administered 2 days later, and a new ovum pickup was performed 24 h afterward. These methods were performed 3 times every 3 days. In control, follicular aspirations occurred at intervals of 1-week without FSH administration o. The aspirated oocytes were evaluated, submitted to in vitro fertilization and the embryos were transferred to the recipients. The average recovery of oocytes was higher (p 0.05) in control cows (12.4±1.8) than in treated cows (9.4±1.3). There was no difference (p>0.05) in the mean percentage of viable oocytes (52.0±3.9 and 62.7±4.7%) or the mean percentage of embryos (41.4±4.8 and 41.5±4.2%) among control and treated cows, respectively. The mean percentage of pregnancy did not differ (p>0.05) for control cows (43.8±2.7%), and treated cows (40.9±6.8%). In conclusion, FSH treatment did not improve oocyte recovery, in vitro fertilization, and pregnancy percentage. However, there is possibility of several consecutive ovum pickup every t3 days, concentrating the in vitro fertilization and the pregnancy percentage.

O presente estudo avaliou vacas Brangus tratadas com doses únicas de hormônio folículo estimulante (FSH) submetidas a aspiração folicular após vinte e quatro horas, para avaliação da recuperação oocitária, fertilização in vitro e taxa de prenhez. Folículos superiores a três milímetros de diâmetro foram aspirados, 200 mg de FSH foram administrados dois dias depois e uma nova aspiração folicular foi realizada 24 horas após. Esses métodos foram efetivados três vezes a cada três dias. No controle, as aspirações foliculares ocorreram em intervalos de uma semana sem administração de FSH. Os oócitos aspirados foram avaliados, submetidos à fertilização in vitro e os embriões foram transferidos em receptoras. A recuperação média dos oócitos foi superior (p 0,05) nas vacas controle (12,4±1,8) do que nas vacas tratadas (9,4±1,3). Não houve diferença (p>0,05) na porcentagem média de oócitos viáveis (52,0±3,9 e 62,7±4,7%) ou na porcentagem média de embriões (41,4±4,8 e 41,5±4,2%) entre vacas controle e vacas tratadas, respectivamente. A porcentagem média de prenhez não diferiu (p>0,05) para as vacas controle (43,8±2,7%) e as tratadas (40,9±6,8%). Em conclusão, o tratamento com FSH não melhorou a recuperação de oócitos, a fertilização in vitro e o percentual de prenhez. No entanto, existe a possibilidade de várias aspirações foliculares consecutivas a cada três

RESUMO

The present study evaluated Brangus cows treated with single doses of follicle stimulating hormone (FSH) subjected to follicular aspiration after 24 h to assess oocyte recovery, in vitro fertilization and pregnancy rate. Follicles exceeding 3 millimeters in diameter were aspirated, 200 mg of FSH was administered 2 days later, and a new ovum pickup was performed 24 h afterward. These methods were performed 3 times every 3 days. In control, follicular aspirations occurred at intervals of 1-week without FSH administration o. The aspirated oocytes were evaluated, submitted to in vitro fertilization and the embryos were transferred to the recipients. The average recovery of oocytes was higher (p<0.05) in control cows (12.4±1.8) than in treated cows (9.4±1.3). There was no difference (p>0.05) in the mean percentage of viable oocytes (52.0±3.9 and 62.7±4.7%) or the mean percentage of embryos (41.4±4.8 and 41.5±4.2%) among control and treated cows, respectively. The mean percentage of pregnancy did not differ (p>0.05) for control cows (43.8±2.7%), and treated cows (40.9±6.8%). In conclusion, FSH treatment did not improve oocyte recovery, in vitro fertilization, and pregnancy percentage. However, there is possibility of several consecutive ovum pickup every t3 days, concentrating the in vitro fertilization and the pregnancy percentage.

O presente estudo avaliou vacas Brangus tratadas com doses únicas de hormônio folículo estimulante (FSH) submetidas a aspiração folicular após vinte e quatro horas, para avaliação da recuperação oocitária, fertilização in vitro e taxa de prenhez. Folículos superiores a três milímetros de diâmetro foram aspirados, 200 mg de FSH foram administrados dois dias depois e uma nova aspiração folicular foi realizada 24 horas após. Esses métodos foram efetivados três vezes a cada três dias. No controle, as aspirações foliculares ocorreram em intervalos de uma semana sem administração de FSH. Os oócitos aspirados foram avaliados, submetidos à fertilização in vitro e os embriões foram transferidos em receptoras. A recuperação média dos oócitos foi superior (p<0,05) nas vacas controle (12,4±1,8) do que nas vacas tratadas (9,4±1,3). Não houve diferença (p>0,05) na porcentagem média de oócitos viáveis (52,0±3,9 e 62,7±4,7%) ou na porcentagem média de embriões (41,4±4,8 e 41,5±4,2%) entre vacas controle e vacas tratadas, respectivamente. A porcentagem média de prenhez não diferiu (p>0,05) para as vacas controle (43,8±2,7%) e as tratadas (40,9±6,8%). Em conclusão, o tratamento com FSH não melhorou a recuperação de oócitos, a fertilização in vitro e o percentual de prenhez. No entanto, existe a possibilidade de várias aspirações foliculares consecutivas a cada três

RESUMO

In Vitro Embryo Production (IVP) is widely used to improve the reproductive efficiency of livestock animals, however increasing the embryo development rates and pregnancy outcomes is still a challenge for some species. Thus, the lack of biological knowledge hinders developing specie-specific IVP protocols. Therefore, the contributions of RNA-seq to generate relevant biological knowledge and improve the efficiency of IVP in livestock animals are reviewed herein.(AU)

Assuntos

RESUMO

In Vitro Embryo Production (IVP) is widely used to improve the reproductive efficiency of livestock animals, however increasing the embryo development rates and pregnancy outcomes is still a challenge for some species. Thus, the lack of biological knowledge hinders developing specie-specific IVP protocols. Therefore, the contributions of RNA-seq to generate relevant biological knowledge and improve the efficiency of IVP in livestock animals are reviewed herein.

Assuntos

RESUMO

The present study evaluated Brangus cows treated with single doses of follicle stimulating hormone (FSH) subjected to follicular aspiration after 24 h to assess oocyte recovery, in vitro fertilization and pregnancy rate. Follicles exceeding 3 millimeters in diameter were aspirated, 200 mg of FSH was administered 2 days later, and a new ovum pickup was performed 24 h afterward. These methods were performed 3 times every 3 days. In control, follicular aspirations occurred at intervals of 1-week without FSH administration o. The aspirated oocytes were evaluated, submitted to in vitro fertilization and the embryos were transferred to the recipients. The average recovery of oocytes was higher (p<0.05) in control cows (12.4±1.8) than in treated cows (9.4±1.3). There was no difference (p>0.05) in the mean percentage of viable oocytes (52.0±3.9 and 62.7±4.7%) or the mean percentage of embryos (41.4±4.8 and 41.5±4.2%) among control and treated cows, respectively. The mean percentage of pregnancy did not differ (p>0.05) for control cows (43.8±2.7%), and treated cows (40.9±6.8%). In conclusion, FSH treatment did not improve oocyte recovery, in vitro fertilization, and pregnancy percentage. However, there is possibility of several consecutive ovum pickup every t3 days, concentrating the in vitro fertilization and the pregnancy percentage.

O presente estudo avaliou vacas Brangus tratadas com doses únicas de hormônio folículo estimulante (FSH) submetidas a aspiração folicular após vinte e quatro horas, para avaliação da recuperação oocitária, fertilização in vitro e taxa de prenhez. Folículos superiores a três milímetros de diâmetro foram aspirados, 200 mg de FSH foram administrados dois dias depois e uma nova aspiração folicular foi realizada 24 horas após. Esses métodos foram efetivados três vezes a cada três dias. No controle, as aspirações foliculares ocorreram em intervalos de uma semana sem administração de FSH. Os oócitos aspirados foram avaliados, submetidos à fertilização in vitro e os embriões foram transferidos em receptoras. A recuperação média dos oócitos foi superior (p<0,05) nas vacas controle (12,4±1,8) do que nas vacas tratadas (9,4±1,3). Não houve diferença (p>0,05) na porcentagem média de oócitos viáveis (52,0±3,9 e 62,7±4,7%) ou na porcentagem média de embriões (41,4±4,8 e 41,5±4,2%) entre vacas controle e vacas tratadas, respectivamente. A porcentagem média de prenhez não diferiu (p>0,05) para as vacas controle (43,8±2,7%) e as tratadas (40,9±6,8%). Em conclusão, o tratamento com FSH não melhorou a recuperação de oócitos, a fertilização in vitro e o percentual de prenhez. No entanto, existe a possibilidade de várias aspirações foliculares consecutivas a cada três

RESUMO

The present study evaluated Brangus cows treated with single doses of follicle stimulating hormone (FSH) subjected to follicular aspiration after 24 h to assess oocyte recovery, in vitro fertilization and pregnancy rate. Follicles exceeding 3 millimeters in diameter were aspirated, 200 mg of FSH was administered 2 days later, and a new ovum pickup was performed 24 h afterward. These methods were performed 3 times every 3 days. In control, follicular aspirations occurred at intervals of 1-week without FSH administration o. The aspirated oocytes were evaluated, submitted to in v itrofertilization and the embryos were transferred to the recipients. The average recovery of oocytes was higher (p<0.05) in control cows (12.4±1.8) than in treated cows (9.4±1.3). There was no difference (p>0.05) in the mean percentage of viable oocytes (52.0±3.9 and 62.7±4.7%) or the mean percentage of embryos (41.4±4.8 and 41.5±4.2%) among control and treated cows, respectively. The mean percentage of pregnancy did not differ (p>0.05) for control cows (43.8±2.7%), and treated cows (40.9±6.8%). In conclusion, FSH treatment did not improve oocyte recovery, in vitro fertilization, and pregnancy percentage. However, there is possibility of several consecutive ovum pickup every t3 days, concentrating the in vitro fertilization and the pregnancy percentage.

O presente estudo avaliou vacas Brangus tratadas com doses únicas de hormônio folículo estimulante (FSH) submetidas a aspiração folicular após vinte e quatro horas, para avaliação da recuperação oocitária, fertilização in vitro e taxa de prenhez. Folículos superiores a três milímetros de diâmetro foram aspirados, 200 mg de FSH foram administrados dois dias depois e uma nova aspiração folicular foi realizada 24 horas após. Esses métodos foram efetivados três vezes a cada três dias. No controle, as aspirações foliculares ocorreram em intervalos de uma semana sem administração de FSH. Os oócitos aspirados foram avaliados, submetidos à fertilização in vitro e os embriões foram transferidos em receptoras. A recuperação média dos oócitos foi superior (p<0,05) nas vacas controle (12,4±1,8) do que nas vacas tratadas (9,4±1,3). Não houve diferença (p>0,05) na porcentagem média de oócitos viáveis (52,0±3,9 e 62,7±4,7%) ou na porcentagem média de embriões (41,4±4,8 e 41,5±4,2%) entre vacas controle e vacas tratadas, respectivamente. A porcentagem média de prenhez não diferiu (p>0,05) para as vacas controle (43,8±2,7%) e as tratadas (40,9±6,8%). Em conclusão, o tratamento com FSH não melhorou a recuperação de oócitos, a fertilização in vitro e o percentual de prenhez. No entanto, existe a possibilidade de várias aspirações foliculares consecutivas a cada três dias, concentrando a fertilização in vitro e o percentual de prenhez.

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PURPOSE: To evaluate the effects of this thymol-rich oil in the proliferation of human adipose tissue-derived stem cells. METHODS: Stem cells were isolated from human adipose tissue by liposuction. After the first passage, cells were cultivated in triplicate for three days in control medium and medium supplemented with three oil samples (1.0 µg/mL, 5.0 µg/mL, and 25.0 µg/mL). Cells were analyzed by the MTT assay at passage 1 (P1), and cell proliferation of control and 1 µg/mL groups was determined with a hemocytometer at P2 and P3. RESULTS: Viability of the essential oil-treated cells was significantly higher than the control group at P1 (p = 0.0008). The treatment with the oil, at a concentration of 1 µg/mL, led to increases of 24.8% at P1 and 43.0% at P3 in the rate of cell proliferation compared with control cells. CONCLUSION: Supplementing culture medium with essential oil of Lippia origanoides increased cell proliferation, especially at later passages.

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Abstract Purpose: To evaluate the effects of this thymol-rich oil in the proliferation of human adipose tissue-derived stem cells. Methods: Stem cells were isolated from human adipose tissue by liposuction. After the first passage, cells were cultivated in triplicate for three days in control medium and medium supplemented with three oil samples (1.0 μg/mL, 5.0 μg/mL, and 25.0 μg/mL). Cells were analyzed by the MTT assay at passage 1 (P1), and cell proliferation of control and 1 μg/mL groups was determined with a hemocytometer at P2 and P3. Results: Viability of the essential oil-treated cells was significantly higher than the control group at P1 (p = 0.0008). The treatment with the oil, at a concentration of 1 µg/mL, led to increases of 24.8% at P1 and 43.0% at P3 in the rate of cell proliferation compared with control cells. Conclusion: Supplementing culture medium with essential oil of Lippia origanoides increased cell proliferation, especially at later passages.

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Purpose: To evaluate the effects of this thymol-rich oil in the proliferation of human adipose tissue-derived stem cells. Methods: Stem cells were isolated from human adipose tissue by liposuction. After the first passage, cells were cultivated in triplicate for three days in control medium and medium supplemented with three oil samples (1.0 g/mL, 5.0 g/mL, and 25.0 g/mL). Cells were analyzed by the MTT assay at passage 1 (P1), and cell proliferation of control and 1 g/mL groups was determined with a hemocytometer at P2 and P3. Results: Viability of the essential oil-treated cells was significantly higher than the control group at P1 (p = 0.0008). The treatment with the oil, at a concentration of 1 µg/mL, led to increases of 24.8% at P1 and 43.0% at P3 in the rate of cell proliferation compared with control cells. Conclusion: Supplementing culture medium with essential oil of Lippia origanoides increased cell proliferation, especially at later passages.(AU)

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A produção de búfalos está se expandindo na Amazônia, sendo assim o uso de biotecnologias reprodutivas em programas de melhoramento genético tornou-se uma estratégia para melhorar a produtividade dos rebanhos sem a necessidade de aumentar as áreas de pastagens. A Produção In Vitro de Embriões (PIVE) é uma das biotecnologias disponíveis, a qual é contiuamente aperfeiçoada através da pesquisa de novos componentes para os meios de cultivo e da pesquisa de protocolos espécie específicos que resultem em maior número de embriões de boa qualidade. Contudo, a falta de informações sobre os requerimentos metabólicos de gametas e embriões têm dificultado a consolidação de protocolos de PIVE específicos para búfalos. Neste artigo discutimos as abordagens transcriptômicas que estão ajudando a preencher esta falta de informações, as limitações destas abordagens e os tópicos de pesquisa que ainda precisam ser investigados em búfalos.

Buffalo production is expanding in the Amazon region therefore the use of reproductive biotechnologies in breeding programs became a strategy to improve the herd productivity without a need for increasing pasture areas. In Vitro Production (IVP) is one of the available biotechnologies, which is continuously being improved through the research of novel components of culture media and the research of specie-specific protocols to obtain an increased number of good quality embryos. However, the lack of information about the metabolic requirements of gametes and embryos have delayed the consolidation of buffalo’s IVP specific protocols. Herein, we discussed the transcriptomic approaches that are helping to fill the lack of information, the limitations of these approaches and the research topics that still needs further investigations in buffaloes.

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RESUMO

A produção de búfalos está se expandindo na Amazônia, sendo assim o uso de biotecnologias reprodutivas em programas de melhoramento genético tornou-se uma estratégia para melhorar a produtividade dos rebanhos sem a necessidade de aumentar as áreas de pastagens. A Produção In Vitro de Embriões (PIVE) é uma das biotecnologias disponíveis, a qual é contiuamente aperfeiçoada através da pesquisa de novos componentes para os meios de cultivo e da pesquisa de protocolos espécie específicos que resultem em maior número de embriões de boa qualidade. Contudo, a falta de informações sobre os requerimentos metabólicos de gametas e embriões têm dificultado a consolidação de protocolos de PIVE específicos para búfalos. Neste artigo discutimos as abordagens transcriptômicas que estão ajudando a preencher esta falta de informações, as limitações destas abordagens e os tópicos de pesquisa que ainda precisam ser investigados em búfalos.(AU)

Buffalo production is expanding in the Amazon region therefore the use of reproductive biotechnologies in breeding programs became a strategy to improve the herd productivity without a need for increasing pasture areas. In Vitro Production (IVP) is one of the available biotechnologies, which is continuously being improved through the research of novel components of culture media and the research of specie-specific protocols to obtain an increased number of good quality embryos. However, the lack of information about the metabolic requirements of gametes and embryos have delayed the consolidation of buffalos IVP specific protocols. Herein, we discussed the transcriptomic approaches that are helping to fill the lack of information, the limitations of these approaches and the research topics that still needs further investigations in buffaloes.(AU)

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