Application of highly sensitive saturation labeling to the analysis of differential protein expression in infected ticks from limited samples

Table 1 Rhipicephalus spp. ticks naturally infected with Rickettsia, Ehrlichia, Theileria or Anaplasma species.

Tick species	Collection	Pathogen infection	N ^a)	Total proteins extracted (μg) b)
R. sanguineus	questing	R. conorii	3	57.9
R. sanguineus	dog	E. canis	2	40.1
R. bursa	sheep	T. annulata	9	92.2
R. turanicus	sheep	A. ovis	2	18.5

Questing and feeding Rhipicephalus spp. adult female ticks were collected in Sicilian farms and analyzed for pathogen infection by PCR or RLB. To define pathogen species infecting ticks, PCR and sequence analysis of cloned amplicons were performed for Anaplasma, Ehrlichia and Rickettsia spp. For Theileria spp., RLB results were confirmed at the species level. For proteomics analysis, sex and collection-matching uninfected controls were used. Uninfected ticks were negative for all pathogens analyzed.
^a) Number of ticks comprising the sample pool for 2-DE.
^b) After protein extraction with TriReagent (Sigma), interfering components for 2D-DIGE experiments were removed by using 2D-Clean up Kit (GE Healthcare) and protein concentration was determined using the 2D-Quant Kit (GE Healthcare). For analysis, protein samples were pooled and table shows the total quantity obtained for each group.

ISSN: 1477-5956