AUTHOR=Salzano Angela , Russo Marco , Anglani Giuseppe , Licitra Francesca , Zullo Gianluigi , Cotticelli Alessio , Fatone Gerardo , Campanile Giuseppe 

TITLE=Early Prediction of Corpus Luteum Functionality Using an Imaging Software

JOURNAL=Frontiers in Veterinary Science

VOLUME=7

YEAR=2020

URL=https://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2020.00299

DOI=10.3389/fvets.2020.00299

ISSN=2297-1769

ABSTRACT=<p>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 (<italic>Bubalus bubalis</italic>). 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 (P<sub>4</sub>) concentrations. Pregnancy diagnosis was carried out on Day 45 by ultrasound and confirmed on Day 70 post-TAI. Differences in CLS, BFA, and P<sub>4</sub> 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 (<italic>n</italic> = 29; 55%) had larger CLS (2.2 ± 0.1 vs. 1.9 ± 0.1 cm<sup>2</sup>; <italic>P</italic> &lt; 0.01), higher BFA (0.6 ± 0.0 vs. 0.4 ± 0.0 cm<sup>2</sup>; <italic>P</italic> &lt; 0.01), and higher P<sub>4</sub> blood level (1.8 ± 0.1 vs. 1.4 ± 0.1; <italic>P</italic> &lt; 0.01) during Day 5–10 as compared to not-pregnant buffaloes (<italic>n</italic> = 22). Throughout the entire period, the first feature that changed between groups was P<sub>4</sub> blood concentration at Day 7 (1.7 ± 0.1 vs. 1.2 ± 0.1; <italic>P</italic> &lt; 0.05) followed by BFA at Day 8 (0.6 ± 0.0 vs. 0.5 ± 0.0; <italic>P</italic> &lt; 0.05), respectively, in pregnant and not-pregnant animals. The ROC analyses indicated that P<sub>4</sub> was able to predict pregnancy since Day 5 (<italic>P</italic> &lt; 0.05) although a more reliable result could be obtained from Day 8 (<italic>P</italic> &lt; 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 P<sub>4</sub> concentration (odds ratio 534.127; <italic>P</italic> &lt; 0.01) and BFA (odds ratio 744.893; <italic>P</italic> &lt; 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.</p>