Immunoproteomic Analysis of Trichinella britovi Proteins Recognized by IgG Antibodies from Meat Juice of Carnivores Naturally Infected with T. britovi

Abstract

Infection with Trichinella nematodes elicits non-specific and specific immune responses; these depend on the dose of infection, the nematode, and the host species. Few studies have examined the presence of specific antibodies against Trichinella spp. in the meat juice of wild animals. The aims of the study were to determine the prevalence of antibodies against Trichinella spp. in meat juice and to identify the specific proteins reacting with the meat juice from free-living carnivores naturally infected with the parasite. Meat juice samples were taken from foxes, badgers, raccoon dogs, and martens and tested with indirect ELISA. Antibodies against Trichinella spp. were detected in 10% of foxes and 46% of raccoon dogs. The ELISA results were confirmed by immunoblot, which revealed different protein patterns in meat juice from red foxes, raccoon dogs, and badgers. The most frequently observed bands were sent for further analysis by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for the detection of Trichinella britovi immunogenic proteins. The results confirm the presence of proteins such as serine protease and heat shock proteins associated with Trichinella infection. These findings provide that meat juice is a useful matrix for proteomic analysis.

Keywords: Trichinella britovi; antibodies; meat juice immunology; wildlife.

Figure 1

The immunoblot analysis of T. britovi ML C antigens incubated with meat juice samples taken from different animals. Lanes 1–3—meat juice samples taken from foxes; lanes 4–15—meat juice samples taken from raccoon dogs; lane 16—meat juice sample taken from a badger; lanes 17–20—meat juice samples taken from badger, marten, fox, and raccoon dog (respectively), which were used as a negative control for immunoblot. Lines marked with + were recognized as positive in ELISA and immunoblot.

Figure 2

(A) The immunoblot analysis of T. britovi ML C antigens incubated with selected meat juice samples. (B) Signal intensity and relative migration values for T. britovi ML C antigens incubated with examined meat juice samples. Lanes 1–2—meat juice samples from foxes (F1, F2, respectively); Lanes 3–5—meat juice samples from raccoon dogs (R1, R2, R3, respectively). No. of bands—A, B, C, D, E, F, G.

Figure 3

The percentage share of the number of proteins derived from three categories: biological process, cellular component, and molecular function.