Abstract
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.
Highlights
The corpus luteum (CL) is essential for establishing and maintaining pregnancy in females [1,2,3]
In terms of differences per each day within the two groups (P and NP), the blood flow area (BFA) and the P4 concentration increased in a time-dependent manner in P group, and in the NP group, the same features started to decrease from Day 9 (Table 1)
We demonstrated that a post-processing tool, such as ImageJ, used to analyze Doppler images could be a suitable tool for studying CL blood flow, whereas the common examinations of the CL area did not seem to be as useful for that purpose
Summary
The corpus luteum (CL) is essential for establishing and maintaining pregnancy in females [1,2,3]. The advent of color Doppler ultrasound imaging made it possible to examine the intensity of CL blood flow [4] and allows for the generation of real-time information about CL functionality in different phases of the estrous cycle. Buffaloes with higher CL vascularization on Day 8 after TAI had a higher pregnancy rate, and there is a positive relationship between TAMV values, P4 concentrations, and pregnancy rates
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