The present study aimed to assess the applicability of luteal blood flow data acquired through the use of color Doppler ultrasonography and a post-processing analysis tool (ImageJ) for predicting pregnancy in buffaloes (Bubalus bubalis). The experiment was carried out on 59 multiparous Italian Mediterranean buffaloes that underwent synchronization of estrus and fixed-time artificial insemination (TAI). Corpus luteum features (size: CLS and blood flow: BFA) were taken from Day 5 to 10 after TAI and retrospectively measured with ImageJ. In the same period, blood samples were taken to assess progesterone (P₄) concentrations. Pregnancy diagnosis was carried out on Day 45 by ultrasound and confirmed on Day 70 post-TAI. Differences in CLS, BFA, and P₄ concentrations from Day 5 to 10 after TAI measured between groups were analyzed by ANOVA repeated measures as were differences within each day of measuring. Buffaloes that established a pregnancy (n = 29; 55%) had larger CLS (2.2 ± 0.1 vs. 1.9 ± 0.1 cm²; P < 0.01), higher BFA (0.6 ± 0.0 vs. 0.4 ± 0.0 cm²; P < 0.01), and higher P₄ blood level (1.8 ± 0.1 vs. 1.4 ± 0.1; P < 0.01) during Day 5–10 as compared to not-pregnant buffaloes (n = 22). Throughout the entire period, the first feature that changed between groups was P₄ blood concentration at Day 7 (1.7 ± 0.1 vs. 1.2 ± 0.1; P < 0.05) followed by BFA at Day 8 (0.6 ± 0.0 vs. 0.5 ± 0.0; P < 0.05), respectively, in pregnant and not-pregnant animals. The ROC analyses indicated that P₄ was able to predict pregnancy since Day 5 (P < 0.05) although a more reliable result could be obtained from Day 8 (P < 0.01). At Day 10, it was possible to set a cutoff value for every parameter taken into account. The logistic regression analysis showed that pregnancy was positively influenced by P₄ concentration (odds ratio 534.127; P < 0.01) and BFA (odds ratio 744.893; P < 0.01). In conclusion, the use of color Doppler ultrasonography, together with ImageJ, identified different patterns of BFA between pregnant and not-pregnant buffaloes starting from Day 8 post-TAI.

本研究旨在评估通过使用彩色多普勒超声检查和后处理分析工具（ImageJ）获得的黄体血流数据的适用性，以预测水牛的妊娠（水牛）。该实验是在59个意大利多头地中海水牛上进行的，这些水牛同步进行了发情和定时人工授精（TAI）。在TAI后第5天到第10天采集黄体特征（大小：CLS，血流量：BFA），并用ImageJ进行回顾性测量。在同一时期，采集血液样本以评估孕酮（P ₄）浓度。妊娠诊断在第45天通过超声进行，并在TAI后第70天确认。通过ANOVA重复测量分析各组之间在TAI后第5天至第10天的CLS，BFA和P ₄浓度的差异，以及每天测量的差异。确定怀孕的水牛（ñ= 29; 55％）的CLS较大（2.2±0.1 vs. 1.9±0.1 cm ²；P<0.01），更高的BFA（0.6±0.0对0.4±0.0 cm ² ;P<0.01），以及更高的P ₄血液水平（1.8±0.1 vs. 1.4±0.1;P 与未怀孕的水牛相比，在第5-10天的时间<0.01）ñ= 22）。在整个期间，两组之间的第一个变化特征是第7天的P ₄血药浓度（1.7±0.1对1.2±0.1；P <0.05），然后是第8天的BFA（0.6±0.0对0.5±0.0； P<0.05），分别在怀孕和未怀孕的动物中。ROC分析表明，从第5天起，P ₄就能预测怀孕（P <0.05），尽管从第8天起可以获得更可靠的结果（P<0.01）。在第10天，可以为所考虑的每个参数设置一个截止值。Logistic回归分析显示，怀孕受到P ₄浓度的正影响（比值534.127；P <0.01）和BFA（比值744.893; P<0.01）。总之，彩色多普勒超声检查与ImageJ的结合使用可从TAI后第8天开始，在孕妇和非孕妇水牛之间发现不同的BFA模式。