Abstract
Ovarian follicle stimulation with exogenous FSH before ovum pickup (OPU) in Bos taurus females is a common practice to increase invitro embryo production (IVP). The optimal stimulatory period length for OPU-IVP, however, has not been definitively ascertained. The objective of the present study, therefore, was to determine the effect of length of the superstimulatory treatment period before OPU on ovarian response and IVP in Holstein heifers. Nonpregnant heifers (n=57) 13.8±0.2 months of age with moderate body condition score (3.0±0.1; scale 1 to 5) were assigned in a completely randomised design to one of the following experimental groups: FSH2d: 200mg of p-FSH (Folltropin-V®, Vetoquinol) distributed in four injections (60, 60, 40, and 40mg) of FSH 12h apart; FSH3d: 200mg of p-FSH distributed in six injections (40, 40, 40, 40, 20, and 20mg) of FSH 12h apart. Timing of follicular wave emergence was synchronized by dominant follicle removal 36h before the first p-FSH injection in all heifers. An intravaginal progesterone (P4) implant (1.38g of P4 CIDR®, Zoetis) was inserted at the time of the first p-FSH injection and removed at the time of OPU, which occurred in all heifers at 44h (Nivet et al. 2012 Reproduction 143, 165-171; https://doi.org/10.1530/REP-11-0391) after the last p-FSH injection. Additionally, follicle number was determined at OPU and classified as small (<6mm), medium (6–10mm) or large (>10mm). Oocytes from follicles of different sizes were pooled by heifer at OPU and then classified and subjected to IVP procedures. Differences between treatment groups were evaluated using generalized linear mixed models (SAS 9.4; SAS Institute Inc.) and data are presented as mean±s.e.m. (Table 1). Lengthening the FSH treatment period resulted in a greater (P=0.01) number of large follicles; however, the number of small, medium-size, and total follicles was not different between groups. Number of total recovered oocytes, viable oocytes, cleaved oocytes, as well as recovery rate, percent viable oocytes, and cleavage rate were not different (P>0.2) between groups. Similarly, the number of blastocysts produced per heifer and blastocyst rate were not different (P>0.9) among groups. In conclusion, lengthening the period of FSH treatment by 1 day increased the number of large follicles at OPU but did not improve overall ovarian response, oocyte recovery, or embryo production. Table 1. Ovarian response and IVP of heifers treated with p-FSH during 2 or 3 days before ovum pickup Response FSH2d (n=28) FSH3d (n=29) P-value Small follicles (n) 5.9±0.6 5.7±0.8 0.83 Medium follicles (n) 17.0±2.4 12.9±1.6 0.18 Large follicles (n) 2.5±0.5 4.5±0.6 0.01 Total follicles (n) 25.4±2.6 23.1±1.8 0.60 Total oocytes (n) 17.0±2.5 13.0±1.2 0.23 Recovery rate (%) 62.6±3.7 56.9±3.1 0.26 Viable oocytes (n) 14.4±2.0 11.3±1.1 0.30 Viable oocytes (%) 85.0±2.4 88.0±3.0 0.31 Cleaved oocytes (n) 8.7±1.6 7.1±1.2 0.62 Cleavage rate (%) 54.7±5.7 54.1±5.7 0.96 Blastocysts (n) 3.2±0.6 3.2±0.7 0.98 Blastocyst rate (%) 20.6±4.0 22.4±3.7 0.97
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