116 Anti-Mullerian hormone in selecting donors for ovum pickup in cattle: generating a cutoff using ROC analysis

Choosing the right donors is crucial for the success of ovum pickup (OPU), in vitro embryo production (IVEP), and embryo transfer (ET) programs. Understanding the follicular pool can improve oocyte recovery and embryo production per donor in their lifetime. However, using ultrasound machines for this purpose is often difficult in field conditions, and per-rectal examination is inaccurate in making the judgment. Hence, a biomarker for the follicular pool is required. Anti-Mullerian hormone (AMH) has a strong correlation with antral follicle count and the number of oocytes retrieved through OPU in cattle. However, to use AMH for selecting donors, reference values and cutoffs must be established. Therefore, the aim of this study was to determine AMH cutoff values for selecting donors in Bos indicus cattle for OPU. Blood samples were collected from 41 donors (Gir, Kankrej, Red Sindhi, Sahiwal, and Tharparkar) during both the luteal and follicular phases of the estrous cycle. The serums were separated and stored at −20°C until further analysis. AMH levels were measured using ELISA (Cloud Clone Corp.). For each animal, the average AMH levels during the luteal and follicular phases were calculated for subsequent analysis. Donors were classified based on average oocyte recovery (14.7 ± 8.1; mean ± SD). Animals with an average oocyte recovery greater than 22.8 were assigned to the high group (n = 8). Those with an average oocyte recovery of less than 6.6 were placed in the low group (n = 5). The remaining animals were placed in the moderate group (n = 28). AMH levels were calculated for each group, with means reported as mean ± s.e.m. (pg mL⁻¹). Comparisons between groups were made using Student’s t-test, with significance set at P < 0.05. AMH values were further analyzed using the receiver operating characteristic (ROC) curve, and the cutoff value was determined based on the optimal balance of sensitivity and specificity. On average, animals presented AMH concentrations of 2682.6 ± 274.0 pg mL⁻¹ (ranging from 618.6 to 10 632.3). No significant difference (P = 0.88) in AMH values was observed between the samples collected during the luteal (2710.4 ± 289.3 pg mL⁻¹) and follicular phases (2654.7 ± 261.8 pg mL⁻¹), indicating that samples can be collected at any point of the estrous cycle. The AMH values obtained in the high group (2864.8 ± 378.3) were significantly higher (P = 0.04) than those in the low group (1591.1 ± 387.1). The area under the ROC curve for the AMH level was 0.668 (P = 0.0143). Based on the ROC analysis, the optimal cutoff value for AMH is 2020.1 pg mL⁻¹; this value enables deciding whether a donor is good or not based on the AMH value. Sensitivity at this cutoff is 0.80, and specificity at this cutoff is 0.57. The results indicate that AMH positively correlates with oocyte recovery in OPU for cattle. An AMH cutoff of 2020.1 pg mL⁻¹ can be used to identify animals with low oocyte yields in Bos indicus, suggesting that AMH could serve as a valuable biomarker for improving donor selection. This study establishes a cutoff for Bos indicus, but a more detailed analysis with a larger population of specific breeds may be necessary for a more precise selection for individual breeds.