Temporal analysis of early pregnancy related gene transcripts in Murrah Buffaloes

A crucial requirement for effective reproductive management in cattle like cows and buffaloes is the early and accurate pregnancy detection. By allowing the farmer to quickly spot non-pregnant animals and cure and/or rebreed them, early pregnancy identification is essential for reducing the calving interval. The goal of the current study was to standardize the expression of the CCL8 and CXCL10 genes as an early pregnancy marker in Murrah buffaloes. Blood samples were taken on day 16 for gene expression following artificial insemination, where as blood sample collected on day 0, 7, 14, 21 post A.I. for progesterone concentration. Buffaloes were split into two groups, pregnant (n = 6) and non-pregnant (n = 6), based on the day of the resumption of estrus. Using qRT-PCR based on SYBR green dye, the gene expression levels in peripheral blood leucocytes (PBLs) were assessed. CCL8, CXCL10, and GAPDH gene amplification products produced amplicons with respective sizes of 108, 117, and 158 bp. The results of the qPCR analysis demonstrate that CCL8 mRNA gene expression in pregnant Murrah buffaloes was found to be 5.13 and 12.21 fold higher in comparison to non-pregnant Murrah buffaloes, while CXCL10 mRNA expression was found to be 4.19 and 22.17 fold higher in comparison to non-pregnant Murrah buffaloes. As a result, on day 16 of pregnancy, pregnant buffaloes had higher levels of CCL8 and CXCL10 mRNA expression in PBLs than non-pregnant buffaloes. Progesterone levels in the pregnant group significantly rose (p0.05) from day 0 to day 21. On days 0, 7, and 14, however, there was no discernible difference between the pregnant and non-pregnant groups. On day 45 after the A.I., per-rectal examination further verified the pregnancy. Hence, on day 16 gene expression profiling of CCL8 and CXCL10 in Murrah buffaloes may be employed as an early pregnancy marker.