In vivo and in vitro expression of five genes involved in Corynebacterium pseudotuberculosis virulence

Caseous lymphadenitis (LC) is a chronic contagious disease caused by Corynebacterium pseudotuberculosis, which mainly affects goats and sheep. Vaccination is an effective but not yet well-established method, partly due to a lack of knowledge surrounding the most effective immunoprotective components. The present study aimed to quantify and compare the in vivo expression of genes pld (phospholipase D), cpp (CP40), nanH (neuraminidase H), sodC (superoxide dismutase C) and spaC (adhesin) using qRT-PCR, with the respective expression in vitro. Caseous material of abscesses removed from five animals was cultured, with colonies suggestive of C. pseudotuberculosis identified. RNA extraction was performed on these samples, as well as on the respective pellets derived from liquid cultures brain heart infusion. After evaluating RNA integrity, complementary DNA was synthesized, followed by the relative quantification each of the genes of interest. Mean mRNA expression of the five genes found in abscesses and in cultures differed significantly, with respective values of: nanH 811.50 +/- 198.27 and 359.35 +/- 75.45 (p = 0.009); cpp 856.31 +/- 385.11 and 154.54 +/- 94.34 (p = 0.0039); plD 922.70 +/- 450.73 and 212.41 +/- 153.10 (p = 0.016); sodC 1,293.53 +/- 564.75 and 223.63 +/- 145.58 (p = 0.016); spaC 1,157.10 +/- 525.13 and 214.26 +/- 125.70 (p = 0,016). Expression was observed to be 6-8 times higher in abscesses than in cultures, Indicative that is a genetic expression of the in vitro bacterium exists, yet in vivo has a greater magnitude corroborating to one of these virulence factors in the pathogenesis of LC.