Mastitis is a condition in which the mammary tissue becomes inflamed. Changes in color and the appearance of clots, as well as increases in cell counts in the milk, are all indicators of inflammation. Mastitis is a common occurrence in cows as a result of inframammary infections. The present study aimed to find out how often nontuberculous mycobacteria (NTM) mastitis occurs and how hp65 affects Interleukin (IL) 6 concentrations. The findings of the Modified Whiteside Test (MWT) on the milk samples from 70 cows, 50 sheep, and 30 goats revealed that 82.9%, 76.7%, and 46.7% of milk samples from cows, sheep, and goats were positive, respectively. This test demonstrated a range of positive milk sample MWT reactions, and the difference among the current positivity score results was statistically significant (P<0.05). The presence of NTM in analyzed milk samples from cows and sheep was confirmed by hsp65-based polymerase chain reaction (PCR) and gene sequencing, with significant differences (P<0.05) in 71.4% and 20% of milk samples from cows and sheep, respectively. The PCR detection of the NTM hsp65 gene in fecal samples from cows, sheep, and goats indicated that cows (80%) had the highest proportion of NTM hsp65 gene amplification, followed by goats (70%), while sheep fecal samples had the lowest amount (22%). The difference among the positive NTM hsp65-based PCR was statistically significant (P<0.05). The phylogenetic tree and sequence analysis of the hsp65 gene revealed two novel variant NTM hsp65 genes that were deposited in Gene Bank (GenBank acc. LC636294 and LC636295). The current examined NTM Hsp65 Mycobacterium sequences which were included in the Mycobacterium avium clade in the currently produced tree ELISA detection of IL6 concentration in cow's milk revealed that IL-6 concentration in mastitis milk was varied. The mean of IL-6 concentration in cow's mastitis milk with MWT scores (+++ve) and mean of IL6 concentration in each MWT scores (++ve), MWT scores (+ve), and -ve MWT cow's milk had a highly significant difference (P<0.001).
Keywords: IL6; Non-tuberculous mycobacteria; Polymerase chain reaction; hsp65.