Investigating the expression of IRAK1 and TRAF6 genes in the mammary tissue of Holstein cows with clinical mastitis

The objective of this investigation was to compare the relative expression level of the IRAK1 and TRAF6 genes in the mammary tissue of cows afflicted with clinical mastitis and unaffected cows utilizing the Real-time PCR technique. For this particular purpose, a total of 15 mammary tissue samples, consisting of 5 healthy specimens and 10 samples with clinical mastitis. An RNA extraction kit was employed to extract RNA from the designated samples. Following the assessment of both the quality and quantity of the extracted RNA, it was utilized for cDNA synthesis. Subsequently, a pair of specific primers were employed twice to ascertain the relative expression level of the desired genes in relation to the I8SrRNA gene serving as an internal control through the qPCR technique. The relative expression level of the genes was determined using the 2-ΔΔct method, while the statistical discrepancy between the diseased and healthy groups was analyzed utilizing the t-student test in SAS statistical software version 9/1. The acquired outcomes indicated that the expression levels of the TRAF6 and IRAK1 genes in the diseased samples were 2.53 (P<0.024) and 3.04 (P<0.0012) times higher, respectively. The identification of solutions to develop strategies for the control of mastitis disease is of considerable significance, with the treatment thereof possessing substantial economic rationale for the global dairy cattle industry. The results obtained from this investigation may be employed to enhance comprehension pertaining to the role of TRAF6 and IRAK1 genes within the defense system of mammary glands, as well as to devise novel examinations geared towards the identification of polymorphisms within these genes for application in breeding programs aimed at identifying mastitis-resistant animals.