Oviduct fluid during IVF moderately modulates polyspermy in in vitro-produced goat embryos during the non-breeding season.

The present study determined i) the presence of proteins (oviduct-specific glycoprotein, OVGP1; heat shock protein-70A, HSPA1A; heat shock protein-A8, HSPA8; annexin A1, ANXA1; annexin A5, ANXA5; and myosin-9, MYH9) known to be involved in early reproduction in the oviduct fluid (OF) of anestrous goats; and ii) the functional effect of during IVF on polyspermy modulation and embryonic development. In vitro-matured oocytes were co-cultured with spermatozoa (1.0, 2.0, or 4.0 x 106 cells/mL) for 18 h in SOF medium supplemented with 5 µg/mL of heparin, 4 µg/mL gentamicin, and 10% estrus sheep serum (CTRL1, CTRL2, and CTRL4 groups) or the same medium plus 10% OF (OF1, OF2, and OF4 groups) obtained from anestrus goats. The analysis of OF by western blotting confirmed the presence of the six proteins tested for. The increase in sperm concentration had no effect (P > 0.05) on the penetration rate in any group; however, monospermy rate decreased as sperm concentration was increased in both OF and CTRL. Regardless of the concentration used, when data were pooled, OF supplementation improved (P < 0.05) monospermy and tended (P = 0.057) to enhance IVF efficiency. Additionally, IVF efficiency was higher (P < 0.05) in OF1 than in OF4 [60 ± 13 vs 37 ± 5%). The development capacity was not affected (P > 0.05) by the sperm concentration and OF treatment, and the average values were cleavage (72 ± 2.6%), blastocyst (37 ± 3.0%), blastocyst in relation to the cleaved (51 ± 4.8%), hatched (62 ± 1.2%), and number of cells per blastocyst (174 ± 1.8%). In conclusion, the six proteins analyzed are present in the OF of anestrous goats, and the supplementation of this OF during IVF may modulate the polyspermy incidence and enhance IVF efficiency, especially when 1x106 sperm per mL is used.

Exogenous progestogens differentially alter gene expression of immature cumulus-oocyte complexes in sheep.

This study evaluated the role of progesterone (P4) and medroxyprogesterone acetate (MAP) on the molecular status of immature cumulus-oocyte complexes (COCs) and the implications for oocyte quality in sheep. The number of viable COCs per ewe and the rate of COCs screened for developmental competence by brilliant cresyl blue positive (BCB+) were similar (P > 0.05), respectively, across treatments (P4: 7.7 ± 0.7 and 4.7 ± 1.2; MAP: 5.7 ± 1.0 and 3.5 ± 2.3; and control: 5.7 ± 1.1 and 3.6 ± 2.4). The COCs' gene expression was altered by exogenous progestogens compared with the control group: markers of steroidogenic pathway (FSH receptor [FSHr], LH receptor [LHr], and estradiol receptor α) and of quality (zygote arrest 1, growth differentiation factor 9, and B-cell lymphoma 2) were in abundance in P4 (P < 0.05). In addition, reelin protein (RELN) was downregulated, and Bcl-2 was upregulated in MAP (P < 0.05). In the P4 vs MAP comparison, FSHr, LHr, and RELN genes were upregulated (P < 0.05) in the P4 group. In conclusion, P4 and MAP promoted dissimilar effects on transcriptome profiling of immature BCB-selected COCs, possibly due to the differences in the chemical structure of progestogens and concentrations of serum P4. Exogenous P4 impacted positively on the profile of genes related to oocyte quality.

Exogenous progestogen does not affect first-wave follicle populations and oocyte quality during ovarian stimulation with FSH in sheep.

The effect of short-term administration of medroxyprogesterone acetate (MPA) or natural progesterone (P4) during ovarian stimulation with FSH on oocyte recovery was investigated in Santa Inês ewes. Ewes were treated with an intravaginal sponge containing MPA for 6 d; GnRH was applied 36 h after sponge removal and FSH was given in 3 injections (40, 24, and 16 mg, respectively) every 12 h after (D0, approximate ovulation time). At the first FSH dose, the ewes received either a new MPA sponge (n = 10) or a controlled device for internal release impregnated with P4 (n = 10) or did not receive any device (n = 10). Ovarian dynamics were assessed every 12 h by transrectal ultrasonography from D-3 to D2. Oocytes were recovered by laparoscopic ovum pick-up (LOPU) on D2 and graded by morphologic quality. The number of small, medium, and large follicles at D0 and D2 (ultrasound examinations), number of both follicles aspirated and oocytes recovered at LOPU, recovery rate, and oocyte grade did not differ (P > 0.05) among treatments. Thus, the short-term use of MPA or P4 during ovarian stimulation did not affect the first-wave follicle population or morphologic quality of oocytes. We would suggest that, in this protocol, the use of exogenous progestin is unnecessary.

Colour-Doppler ultrasound imaging as a laparoscopy substitute to count corpora lutea in superovulated sheep.

This study evaluated colour-Doppler ultrasound imaging (UI) as a substitute for laparoscopy to count the corpora lutea (CL) in superovulated sheep. Twenty-five Santa Ines ewes were superovulated three times at 21-day intervals. Corpora lutea were counted by colour-Doppler UI (CLDOPPLER ) 6 days after each superovulation and confirmed by laparoscopy (CLLAP ) 12 hr later. The mean number of CL was similar for both techniques (2.1 ± 2.5 vs. 2.1 ± 2.7 for CLDOPPLER and CLLAP , respectively) with a significant positive correlation (r = .94; r2 =.89). Colour-Doppler UI effectively evaluated the ovarian response in superovulated ewes and efficiently identified animals that did not respond to superovulation.

Uso da somatotropina recombinante bovina em búfalas leiteiras I: produção e composição físico-química do leite / Use of recombinant bovine somatotropin (rbST) in dairy buffaloes I: production and physicochemical composition of milk

Objetivou-se avaliar a influência da somatotropina recombinante bovina (rbST) sobre a produção e os constituintes do leite de búfalas entre 63 e 154 dias em lactação. Foram utilizadas 22 búfalas, distribuídas em dois grupos experimentais: grupo rbST - aplicação de 500 mg de rbST a cada 14 dias; grupo controle - sem aplicação de rbST. A cada sete dias, foi aferida a produção de leite de todas as búfalas e coletada uma amostra para análise físico-química. As variáveis produtivas e as oriundas de análises laboratoriais foram avaliadas como medidas repetidas no tempo, utilizando-se o comando Repeated gerado pelo procedimento GLM do SAS. A média dos parâmetros estudados para os grupos rbST e controle foram, respectivamente: produção de leite - 6,54 vs. 6,68 kg; gordura - 6,31 vs. 6,34%; proteína 3,86 vs. 3,81%; lactose - 4,96 vs. 5,02%; sólidos totais - 16,05 vs. 16,03%; extrato seco desengordurado - 9,75 vs. 9,74%; contagem de células somáticas - 329,90 vs. 171,68 (x 1000/mL); e elecondutividade - 2,87 vs. 2,81mS/cm. A utilização de 500mg de rbST administrados quinzenalmente, entre 63 e 154 dias em lactação não alterou a produção de leite, a proporção dos constituintes e a CCS do leite de búfalas leiteiras.(AU)

This study aimed to evaluate the effect of recombinant bovine somatotropin (rbST) on milk yield and the proportion of buffalo milk components during lactation. Twenty-two buffaloes randomly distributed in two experimental groups were used: Group rbST - application of 500mg rbST every 14 days, between 63 and 154 days in milk (DIM); Control Group - without treatment. Weekly, the milk yield of buffaloes was measured and a sample was collected for physicochemical analysis. The response variables were evaluated as repeated measures, using the Repeated procedure through the GLM procedure of SAS. Means of each variable after rbST and Control were: Milk yield - 6,54 vs. 6,68 kg; Fat - 6,31 vs. 6,34%; Protein - 3,86 vs. 3,81%; Lactose - 4,96 vs. 5,02%; Milk solids - 16,05 vs. 16,03%; Defatted dry matter - 9,75 vs. 9,74%; Somatic Cells Count - 329,90 vs. 171,68 (x 1000/mL); and electrical conductivity- 2,87 vs. 2,81mS/cm. The use of 500mg of rbST administered every two weeks, between 63 and 154 DIM did not affect milk yield, proportion of milk constituents and SCC of dairy buffaloes.(AU)