Non-Tuberculous Mycobacteria hsp۶۵ in Relation to Bovine Subclinical Mastitis and IL۶ Concentration in Cow's Milk

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 hp۶۵ affects Interleukin (IL) ۶ concentrations. The findings of the Modified Whiteside Test (MWT) on the milk samples from ۷۰ cows, ۵۰ sheep, and ۳۰ goats revealed that ۸۲.۹%, ۷۶.۷%, and ۴۶.۷% 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<۰.۰۵). The presence of NTM in analyzed milk samples from cows and sheep was confirmed by hsp۶۵-based polymerase chain reaction (PCR) and gene sequencing, with significant differences (P<۰.۰۵) in ۷۱.۴% and ۲۰% of milk samples from cows and sheep, respectively. The PCR detection of the NTM hsp۶۵ gene in fecal samples from cows, sheep, and goats indicated that cows (۸۰%) had the highest proportion of NTM hsp۶۵ gene amplification, followed by goats (۷۰%), while sheep fecal samples had the lowest amount (۲۲%). The difference among the positive NTM hsp۶۵-based PCR was statistically significant (P<۰.۰۵). The phylogenetic tree and sequence analysis of the hsp۶۵ gene revealed two novel variant NTM hsp۶۵ genes that were deposited in Gene Bank (GenBank acc. LC۶۳۶۲۹۴ and LC۶۳۶۲۹۵). The current examined NTM Hsp۶۵ Mycobacterium sequences which were included in the Mycobacterium avium clade in the currently produced tree ELISA detection of IL۶ concentration in cow's milk revealed that IL-۶ concentration in mastitis milk was varied. The mean of IL-۶ concentration in cow's mastitis milk with MWT scores (+++ve) and mean of IL۶ concentration in each MWT scores (++ve), MWT scores (+ve), and -ve MWT cow's milk had a highly significant difference (P<۰.۰۰۱).

Keywords:

hsp۶۵ , Il۶ , non-tuberculous mycobacteria , Polymerase Chain Reaction