Multistep analysis reveals the relationship between blood indices at the time of ovum pick-up and in vitro embryo production in heifers

Abstract

The inflammatory factors of complete blood count (CBC) are associated with a decrease in the in vitro embryo production (IVP) outcome in women. The relation between the blood indices and in vitro fertilization (IVF) outcomes in bovines remains to be elucidated. Using ovum pick-up (OPU), oocytes were retrieved from heifers (n = 60) and inseminated separately with sperm. The blastocyst formation was recorded on day 7 after insemination for each animal and the blood indices were evaluated at the time of OPU. Then, heifers were classified on the basis of (1) blastocyst formation, cleaved vs. failed, or (2) inflammation, low-grade inflammation (lymphocyte counts > 5.6 × 10⁹/L) vs. no inflammation (lymphocyte counts < 5.6 × 10⁹/L). Oocytes derived from heifers with higher lymphocytes, red blood cells (RBC), platelets, hematocrit, red cell distribution width (RDW-SD) and plateletcrit values and lower monocytes, eosinophils, mean corpuscular hemoglobin (MCH) and MCH concentration (MCHC) successfully developed to the blastocyst stage. Heifers with low-grade inflammation numerically had a higher percentage of blastocyst formation than normal heifers. The principle component analysis (PCA) showed that blastocyst formation had the strongest positive association with RDW-cv and RDW-SD, while having a strong negative association with mean corpuscular volume (MCV), hemoglobin, MCHC and MCH. The PCA determined that the number of grade A COCs and the percentage of COCs reached the cleavage stage had a negative association with white blood cells (WBC), lymphocytes, basophils and monocytes, and a positive correlation with platelet to lymphocyte ratio, platelet distribution width (PDW) and plateletcrit. Network mapping detected close similarities between BFR and RDW-SD, MPV, and lymphocytes. The area under the receiver operating characteristic curve (AUC) identified that, eosinophils (AUC 0.80), RDW-SD (AUC 0.76), monocytes (AUC 0.76) and lymphocytes (AUC 0.76) had a good predictive ability to detect heifers with high OPU-IVP outcome (≥60%). In conclusion, these findings suggest that CBC indices at the time of OPU were associated with the IVF outcome and may be incorporated into protocols for the identification of heifers with high potential for blastocyst formation.

Keywords: Blood indices; Bovine oocytes; Eosinophils; In vitro embryo production; Lymphocytes; RDW.