Resumo
The aim of this study was to evaluate the efficacy of controlled internal drug release (CIDR) and its reuse in treatment of inactive ovaries in buffaloes with subsequent resumption of cyclicity and reduction of inter-calving intervals. This study was conducted on 54 anestrous buffaloes suffering from ovarian inactivity. The animals were treated by new CIDRs and disinfected second used CIDRs for 7 or 14 days and PGF2α, with or without GnRH. The highest estrus induction rate (EIR; 100%), pregnancy rate (PR; 100%) and 1st service conception rate (1st service CR; 83.3%) were achieved with the treatment regime (CIDR 7 days plus i.m. injection of 25 mg of PGF2α in the 6th day plus i.m. injection of 10 µg GnRH in 8th) day followed by the treatment regime (CIDR 14 days plus i.m. injection of PGF2α in the 13th day); where the EIR, PR and 1st service CR were 85.7, 71.4 and 57.1%, respectively. It could be concluded that the use of CIDR 7 days + i.m. injection of 25 mg of PGF2α in the 6th day + i.m. injection of 10 µg GnRH in 8th day is an alternative to restart the ovarian activity in buffalo cows.(AU)
Assuntos
Animais , Preparações Farmacêuticas/análise , Ovário/citologia , Búfalos/classificaçãoResumo
The aim of this study was to evaluate the efficacy of controlled internal drug release (CIDR) and its reuse in treatment of inactive ovaries in buffaloes with subsequent resumption of cyclicity and reduction of inter-calving intervals. This study was conducted on 54 anestrous buffaloes suffering from ovarian inactivity. The animals were treated by new CIDRs and disinfected second used CIDRs for 7 or 14 days and PGF2α, with or without GnRH. The highest estrus induction rate (EIR; 100%), pregnancy rate (PR; 100%) and 1st service conception rate (1st service CR; 83.3%) were achieved with the treatment regime (CIDR 7 days plus i.m. injection of 25 mg of PGF2α in the 6th day plus i.m. injection of 10 µg GnRH in 8th) day followed by the treatment regime (CIDR 14 days plus i.m. injection of PGF2α in the 13th day); where the EIR, PR and 1st service CR were 85.7, 71.4 and 57.1%, respectively. It could be concluded that the use of CIDR 7 days + i.m. injection of 25 mg of PGF2α in the 6th day + i.m. injection of 10 µg GnRH in 8th day is an alternative to restart the ovarian activity in buffalo cows.
Assuntos
Animais , Ovário/citologia , Preparações Farmacêuticas/análise , Búfalos/classificaçãoResumo
The aim of the present study was to improve in vitro maturation and cleavage rates of buffalo oocytes. Good quality oocytes were divided into two experiments. In Experiment 1 oocytes were cultured for 24 h in a CO2 incubator at 38.5oC either in TCM-199, Hams F-10, MEM or FertiCult medium supplemented with either 10% FCS or 0.3% BSA. Experiment 2 was carried out to investigate the effect of different hormones (either 50 µg/ml eCG, 50 µg/ml FSH or 1 mg/ml E2) added to four of the afore mentioned media enriched with 10% FCS at the same culture conditions. Matured oocytes were fertilized in vitro using frozen thawed semen capacitated with heparin and caffeine. The sperm-oocytes were co-cultured for 22 h in BO or TALP medium. The fertilized oocytes were cultured in either BO or TALP medium for an additional five days at the same culture conditions and checked daily for cleavage. Addition of FCS to all media led to a higher maturation rate, without any significant variation, than BSA did (75.6 vs. 71.3%). Although no influence on the maturation rate was observed, addition of eCG to TCM-199, Hams F-10 or MEM media resulted in a non-significant increase of in vitro maturation rate of buffalo oocytes compared to other hormonal additives to the same media. Furthermore, supplementation of maturation media with eCG resulted in a non-significant higher in vitro maturation rate of buffalo oocytes compared to FSH and E2 (80.4% for eCG supplemented media vs. 74.0% and 73.0% for FSH and E2, respectively). There was a non-significant difference in the cleavage rate of buffalo oocytes matured in TCM-199 supplemented with either sera or hormones and fertilized either in BO or TALP medium. However, the highest non-significant cleavage rate was achieved when oocytes were matured in TCM-199 supplemented with eCG and fertilized in TALP medium (50%). The overall cleavage rate was not significantly greater in TALP than in BO medium (33.7 vs. 15.5%). It could be concluded that supplementation of maturation media with FCS and/or eCG could successfully improve IVM rate of buffalo oocytes. Furthermore, high cleavage rate could be achieved when oocytes were matured in TCM-199 supplemented with FCS and eCG and fertilized in TALP medium.(AU)
Assuntos
Clivagem do DNA , Oócitos/citologia , Búfalos , Técnicas de Maturação in Vitro de OócitosResumo
The aim of the present study was to improve in vitro maturation and cleavage rates of buffalo oocytes. Good quality oocytes were divided into two experiments. In Experiment 1 oocytes were cultured for 24 h in a CO2 incubator at 38.5oC either in TCM-199, Hams F-10, MEM or FertiCult medium supplemented with either 10% FCS or 0.3% BSA. Experiment 2 was carried out to investigate the effect of different hormones (either 50 µg/ml eCG, 50 µg/ml FSH or 1 mg/ml E2) added to four of the afore mentioned media enriched with 10% FCS at the same culture conditions. Matured oocytes were fertilized in vitro using frozen thawed semen capacitated with heparin and caffeine. The sperm-oocytes were co-cultured for 22 h in BO or TALP medium. The fertilized oocytes were cultured in either BO or TALP medium for an additional five days at the same culture conditions and checked daily for cleavage. Addition of FCS to all media led to a higher maturation rate, without any significant variation, than BSA did (75.6 vs. 71.3%). Although no influence on the maturation rate was observed, addition of eCG to TCM-199, Hams F-10 or MEM media resulted in a non-significant increase of in vitro maturation rate of buffalo oocytes compared to other hormonal additives to the same media. Furthermore, supplementation of maturation media with eCG resulted in a non-significant higher in vitro maturation rate of buffalo oocytes compared to FSH and E2 (80.4% for eCG supplemented media vs. 74.0% and 73.0% for FSH and E2, respectively). There was a non-significant difference in the cleavage rate of buffalo oocytes matured in TCM-199 supplemented with either sera or hormones and fertilized either in BO or TALP medium. However, the highest non-significant cleavage rate was achieved when oocytes were matured in TCM-199 supplemented with eCG and fertilized in TALP medium (50%). The overall cleavage rate was not significantly greater in TALP than in BO medium (33.7 vs. 15.5%). It could be concluded that supplementation of maturation media with FCS and/or eCG could successfully improve IVM rate of buffalo oocytes. Furthermore, high cleavage rate could be achieved when oocytes were matured in TCM-199 supplemented with FCS and eCG and fertilized in TALP medium.
Assuntos
Clivagem do DNA , Oócitos/citologia , Búfalos , Técnicas de Maturação in Vitro de OócitosResumo
The present study was conducted to investigate the effects of ovarian morphology on oocyte quantity and quality, as well as the effect of preincubated granulosa cells (PGCs) on in vitro maturation of buffalo oocytes and steroid hormone production. A total of 52 ovarian pairs were grouped into 3 types: Type I (with functional corpus luteum), Type II (with regressed corpus luteum), or Type III (without corpus luteum). The number of follicles and oocytes per ovary were documented. The follicles were classified into 3 groups: <2 mm, 2 to 6 mm, and >6 mm. Oocytes were classified according to their morphology into one of 4 grades (A, B, C, or D) and according to their cumulus compactness into 4 groups (>3 layers of cumulus cells, 1 to 3 layers, partial remnants of cumulus cells, or no cumulus cells). Preincubated granulosa cells were used to investigate their steroidogenic potential with in vitro maturation. A greater number of vesicular follicles and aspirated oocytes were found in Type III than in Type II or Type I. The number of Grade A and Grade B oocytes was significantly higher (P < 0.01) in number in Type III ovaries compared to other types. Oocytes with >3 layers of cumulus cells had a higher maturation rate than oocytes with partial remnants or no cumulus cells, but had low maturation rate compared to oocytes with 1 to 3 layers of cumulus cells. Besides to the higher maturation rate in compact than denuded oocytes, there was a higher (P < 0.01) rate in compact or denuded oocytes when cultured in vitro with PGCs than the corresponding oocytes with no PGCs. These maturation rates coincided with a higher (P < 0.05) concentration of estradiol-17β when compact oocytes were cultured with or without PGCs compared to denuded oocytes and a higher (P < 0.05) concentration of progesterone after culture with PGCs for both compact and denuded oocytes compared to oocytes with no PGCs. In summary, buffalo ovaries with no corpus luteum may result in a higher number of follicles and good oocytes than those with a corpus luteum. Oocytes with an intact cumulus had better maturation than those with partial or denuded cumulus although the denuded oocytes improved their meiotic competence when cultured in vitro with PGCs.(AU)
Assuntos
Animais , Bovinos , Ovário/anatomia & histologia , Oócitos/citologia , Células da Granulosa/citologia , Búfalos/classificação , Fertilização in vitro/instrumentaçãoResumo
The present study was conducted to investigate the effects of ovarian morphology on oocyte quantity and quality, as well as the effect of preincubated granulosa cells (PGCs) on in vitro maturation of buffalo oocytes and steroid hormone production. A total of 52 ovarian pairs were grouped into 3 types: Type I (with functional corpus luteum), Type II (with regressed corpus luteum), or Type III (without corpus luteum). The number of follicles and oocytes per ovary were documented. The follicles were classified into 3 groups: 6 mm. Oocytes were classified according to their morphology into one of 4 grades (A, B, C, or D) and according to their cumulus compactness into 4 groups (>3 layers of cumulus cells, 1 to 3 layers, partial remnants of cumulus cells, or no cumulus cells). Preincubated granulosa cells were used to investigate their steroidogenic potential with in vitro maturation. A greater number of vesicular follicles and aspirated oocytes were found in Type III than in Type II or Type I. The number of Grade A and Grade B oocytes was significantly higher (P 3 layers of cumulus cells had a higher maturation rate than oocytes with partial remnants or no cumulus cells, but had low maturation rate compared to oocytes with 1 to 3 layers of cumulus cells. Besides to the higher maturation rate in compact than denuded oocytes, there was a higher (P < 0.01) rate in compact or denuded oocytes when cultured in vitro with PGCs than the corresponding oocytes with no PGCs. These maturation rates coincided with a higher (P < 0.05) concentration of estradiol-17β when compact oocytes were cultured with or without PGCs compared to denuded oocytes and a higher (P < 0.05) concentration of progesterone after culture with PGCs for both compact and denuded oocytes compared to oocytes with no PGCs. In summary, buffalo ovaries with no corpus luteum may result in a higher number of follicles and good oocytes than those with a corpus luteum. Oocytes with an intact cumulus had better maturation than those with partial or denuded cumulus although the denuded oocytes improved their meiotic competence when cultured in vitro with PGCs.