Density measurement of Demodex canis by qPCR and analysis of serum cytokine levels in dogs with different clinical forms of demodicosis

Objectives: To quantify (by qPCR) the density of Demodex canis mites in the skin of dogs with demodicosis and in healthy dogs, as well as measuring the serum concentrations of interleukin (IL)-1 beta, IL-6, IL-8, IL-10, IL-12, and tumour necrosis factor-alfa (TNF-alpha). Methods: Fifty-four dogs were divided into three groups: localized demodicosis (LD, n = 16), generalized demodicosis (GD, n = 22), and control group (CG, n = 16). All dogs were subjected to skin scraping, blood collection, and skin biopsy. DNA extraction was performed and the parasite density was established by qPCR. Serum cytokine concentrations were obtained by flow cytometry. Results: The median number of mites in the skin of the GD (6.2 x 10(4) copies/mu L) and LD dogs (1.2 x 10(4) copies/mu L) was statistically higher than that in the CG dogs (8.7 x 10(2) copies/mu L). Whereas there were no significant differences in median IL-1 beta, IL-8, IL-10, IL-12, and TNF-alpha levels among the study groups, there was a statistically higher IL-6 concentration in the LD dogs than in the healthy dogs. Conclusions: According to our results, qPCR is an effective method for measuring the density of D. canis in the canine integument. In addition, the activation of the acute-phase immune response in localized demodicosis can be induced by IL-6 activity.