In vivo embryo production and recovery in lacaune ewes after imposing a superovulation treatment regimen is related to pFSH dose.

This study was conducted to assess effects of different doses of pFSH on follicular recruitment, superovulatory response, ova/embryo recovery, and embryo yield in lactating ewes. Ewes (n = 24) had a superovulation treatment regimen imposed. All ewes were implanted with a progesterone intravaginal device for 9 d, and administered either 100 (G-100) or 200 (G-200) mg pFSH, proportioned into six doses administered at 12-h intervals, starting 60 h before device removal. At 7 days subsequent to progesterone device removal, there were non-surgical embryo recoveries (NSER) from ewes having three or more corpora lutea. At the time of the first pFSH injection, number of antral follicles were similar (P < 0.05) between ewes in the G-100 and G-200 group, however, there were more 3.1-4.0 mm follicles in ewes of the G-200 than G-100 group at the time of the second pFSH administration. Estrous response and CL number were less (P < 0.05) in ewes of the G-100 (66.7 % and 2.6 ±â€¯0.7) than G-200 (91.7 % and 11.6 ±â€¯1.2) group. There were embryo collections from 100 % and 90.9 % of ewes in the G-100 and G-200 groups, respectively (P > 0.05). Viable embryo numbers and ova/embryo recovery rate were greater (P < 0.05) in ewes of the G-200 (6.9 ±â€¯1.1 and 67.8 %) than G-100 (1.0 ±â€¯0.5 and 27.6 %) group. A dose of 200 mg pFSH was more effective in inducing a superovulatory response and embryo yield after NSER in ewes, however, the 100 mg dose was insufficient for these purposes.

Embryo development is impaired in goats that are treated for hydrometra and subsequently subjected to superovulation.

BACKGROUND: Reproductive efficiency after hydrometra (HD) treatment is usually unsatisfactory. METHODS: To identify mechanisms involved in low reproductive efficiency of HD-treated goats, pluriparous dairy goats treated for HD (n=10, HD) or with no reproductive disorders (n=11, control: CONT) were induced to oestrus and superovulated. Goats were mated with fertile bucks and seven days after oestrus, non-surgical embryo recovery was performed. Embryos were evaluated and gene expression was performed. RESULTS: There were no differences (P>0.05) in sexual behaviour parameters, superovulation response, mean number of retrieved structures and viable embryos between groups; although embryo recovery rate was higher (P=0.01) in CONT group. Structures in delayed stage (8-16 cells) were more frequent (P<0.05) in HD (29 vs 1 per cent) goats, as well as the percentage of advanced embryos was greater (P<0.05) for CONT (59.3 vs 33.3 per cent) goats. However, the expression of genes related to apoptosis (BAX and Bcl-2), trophectoderm differentiation (CDX2) and pluripotency maintenance (NANOG) was not affected (P>0.05) in embryos that reached the morulae and blastocyst stages. CONCLUSION: Although the HD embryos that developed to morula and blastocyst stages showed no change in the expression of genes related to their quality and implantation capacity, overall, embryo development was impaired in HD-treated goats.

Embryo yield and quality are associated with progestogen treatment during superovulation protocol in lactating Lacaune ewes.

This study assessed the effect of progestogen treatment length on ovarian parameters and embryo yield in superovulated Lacaune ewes collected by nonsurgical embryo recovery. Twenty-three lactating ewes were superovulated 30 d apart using a cross-over design. All ewes received 60 mg of MAP intravaginal sponges for 6 (G-6 group) or 9 (G-9 group) d. A total dose of 133 mg pFSH was given in six decreasing doses (twice a day) starting at 60 h before device removal. Ultrasound examination of the ovaries was performed at the first pFSH injection and one day before embryo recovery, which was performed 6-7 d after the onset of estrus. Embryo recovery was conducted only in ewes that expressed estrus and were mated. There was no difference (P > 0.05) in the total number of follicles between G-6 (15.7 ± 1.0) and G-9 (15.6 ± 0.8) at the time of the first pFSH treatment. The percentage of responding donors with ≥3 corpora lutea (CL; 78.2% [18/23] vs 69.5% [16/23]), mean (±SEM) CL number (7.0 ± 1.2 vs 8.1 ± 1.6), transcervical passage rate (94.4% [17/18] vs 83.3% [15/18], and ova/embryo recovery rate (54.5% [60/110] vs 68.0% [83/122]) were not different (P > 0.05) between the G-6 and G-9 groups. However, the mean number of viable embryos was lower (P < 0.05) in the G-6 group (1.8 ± 0.7) than in the G-9 group. (3.5 ± 1.1). In conclusion, treatment with an intravaginal MAP sponge for 9 d during a superovulation protocol is beneficial for viable embryo yield in Lacaune ewes out of the breeding season.

Gene expression patterns of in vivo-derived sheep blastocysts is more affected by vitrification than slow freezing technique.

Transfer of fresh sheep embryos frequently results in higher pregnancy rate compared to cryopreserved ones, possibly due to a failure in the communication between the cryopreserved embryo and the endometrium during pre-implantation and pregnancy establishment. Thus, this study assessed the effect of sheep embryo cryopreservation (slow freezing or vitrification) on embryo survival rate and expression of genes related to trophectoderm differentiation (CDX2), pluripotency maintenance (NANOG), cell proliferation (TGFB1), mitochondrial activity (NRF1) and apoptosis (BAX and BCL2). Superovulation (n = 32 ewes) was performed and embryos were transcervically collected. One hundred good quality (Grade I and II) embryos were allocated into three groups: fresh embryos (CTL; n = 15), slow freezing (SF; n = 42) or vitrification (VT; n = 43). After thawing/warming, three pools of five blastocysts per group were used for RT-qPCR; the remaining 55 embryos were cultured in vitro in SOFaa medium at 38.5 °C and 5% CO2 (SF: n = 27 and VT: n = 28). Survival rate of SF and VT were, respectively, 29.6% (8/27) and 14.2% (4/28) at 24 h; and 48.1% (13/27) and 32.1% (9/28) at 48 h (P > 0.05). Only CDX2 was affected (up-regulated, P < 0.05) in both groups compared to CTL. The BAX transcript was upregulated in VT, compared to SF group. The VT increased (P < 0.05) the expression of all genes, except for NANOG and NRF1, when compared to the CTL. In conclusion, although in vitro survival was similar between techniques, VT led to increased changes in blastocyst gene expression compared to CTL and SF.

Acrodermatitis dysmetabolica as a sign of methylmalonic aciduria decompensation.

Methylmalonic aciduria children must follow an adequate diet with low protein intake and should be regularly monitored to prevent complications. Although skin lesions like acrodermatitis enteropathica are rare in this disease, their appearance should be correlated with possible low plasma isoleucine level and it can be a sign of decompensation.

Transient neonatal diabetes due to a missense mutation (E227K) in the gene encoding the ATP-sensitive potassium channel (KCNJ11).

Neonatal diabetes is a monogenic form of diabetes. Herein, we report on a newborn presenting diabetic ketoacidosis at 17 days of life. A KCNJ11 mutation was identified. In such cases, insulin can be replaced by sulfonylurea with a successful metabolic control, as an example of how molecular diagnosis may influence the clinical management of the disorder.