Abstract
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.
Highlights
Caseous lymphadenitis (CL), a chronic contagious disease that affects small ruminants, is caused by Corynebacterium pseudotuberculosis, a gram-positive, pleomorphic, non-sporulated, facultatively anaerobic bacillus belonging to the Corynebacteriaceae family (Actinomycetales order).The literature is inconsistent with regard to the prevalence of this disease in goat and sheep herds
The strategy to control caseous lymphadenitis is mainly based on vaccination, which has yet to be widely applied throughout the region, in addition to the adoption of complementary measures concerning the inclusion of new animals in herds, early detection of infected animals, segregation, treatment or elimination of diseased animals, special care in routine handling, as well as other measures (Guimarães et al 2011; Windsor 2011; Kumar et al 2013)
Discussion mRNA detection of the nanH, cpp, pld, sodC, and spaC genes in bacterial cells obtained from culture indicated that transcription of these genes occurred, in addition to the possible synthesis of the respective proteins
Summary
Caseous lymphadenitis (CL), a chronic contagious disease that affects small ruminants, is caused by Corynebacterium pseudotuberculosis, a gram-positive, pleomorphic, non-sporulated, facultatively anaerobic bacillus belonging to the Corynebacteriaceae family (Actinomycetales order).The literature is inconsistent with regard to the prevalence of this disease in goat and sheep herds. In Brazilian goats raised in semi-arid areas, studies conducted. Meyer (2003), in an extensive serological investigation involving 1966 goats conducted in 19 municipalities throughout the semi-arid zone of the state of Bahia, found highly variable prevalence, with infection rates ranging from 9.2 to 72.2%. Considering that most goat herds are concentrated in the northeastern region of Brazil, together with the fact that these animals are essential for the sustenance of small producers, caseous lymphadenitis poses a relevant threat to the raising of livestock in this region (Meyer 2003). The strategy to control caseous lymphadenitis is mainly based on vaccination, which has yet to be widely applied throughout the region, in addition to the adoption of complementary measures concerning the inclusion of new animals in herds, early detection of infected animals, segregation, treatment or elimination of diseased animals, special care in routine handling, as well as other measures (Guimarães et al 2011; Windsor 2011; Kumar et al 2013)
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