الفهرس 
REVIEW OF LITERATUREOnly 14 pages are availabe for public view
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Abstract
Postpartum endometritis is one of the most prevalent illnesses in dairy animals, particularly cattle and buffalo, resulting in significant economic losses due to increased inter-calving intervals. The fertility deficiency also leads to the decrease of buffalo’s production. Determining the immunological state of buffalo relative to the development of endometritis could help improve some reproductive management strategies and reduce the economic loss of buffalo production.
Furthermore, the early diagnosis of subclinical endometritis is enhanced by investigation of differences in the expression patterns of immune-related genes. In this respect, the development of clinical or subclinical endometritis in buffalo was linked to the expression of inflammatory-related genes in uterine tissue.
Immune genes linked to reproductive diseases can be distinguished by how they are expressed in high and low responders. Cytokines are proteins produced spontaneously by immune cells that play a major role in the host’s defense against infection as well as specific and nonspecific immunity. The type of cytokine secreted by a cell is determined by the antigenic stimulation and the type of activated cells.
Accordingly, the purpose of this study was to investigate the gene expression of different immune genes in endometritis-infected buffaloes by using Real Time PCR amplification. RNA was extracted from uteri samples taken from 120 Egyptian buffaloes, 60 animals had endometritis signs including abnormal secretions as well as symptoms of inflammation in the uterus, such as swelling, redness, and hardness; while the other 60 animals had healthy appearance.
QRT-PCR was performed on cDNA, synthesized from extracted RNA, using Sybr green and GAPDH as a house-keeping gene. The analysis is based on the mean cycle threshold (Ct) values of triplicate samples. Using GAPDH as a housekeeping gene for normalization, the Chi-square test was employed to examine the significant differences (P<0.05) in gene expression of the tested genes.
The melting curves for the ten tested genes, IL-1α, IL-1β, IL-6, IL-10, TNF-α, TGFBR1, PTGER2, PTGER4, HP and CXCL5, as well as the housekeeping gene GAPDH, showed a single peak, indicating the absence of primer-dimer formation and specificity of the amplification.
The current findings indicate that the ten genes had different responses toward endometritis infection in buffaloes. mRNA expression of five of these genes: IL-1α, IL-1β, IL-6, TGFBR and CXCL5 were up-regulated with fold changes of 1.3, 1.7, 5, 56.1 and 4.3, respectively, comparing to healthy animals. On the contrary, the expression of the other five genes: IL-10, TNF-α, HP, PTGER2 and PTGER4 were down-regulated with fold changes of 0.2, 0.4, 0.6, 0.6 and 0.4, respectively, in infected buffaloes compared with those in healthy animals.
The present results indicated that the relative expression of inflammation-related immunity genes may have an effective role on the early detection of endometritis infection in buffalo and this early diagnosis can reduce the economic loss of buffalo production and reproduction.
Finally, the mRNA expression of the studied ten genes are found to be related to buffalo uterine health status and differ between infected and healthy animals. So, it can be concluded that the investigation of immunity gene’s expression using real time PCR can be used as diagnostic tools especially for subclinical endometritis infection in buffalo.