Tick-borne disease agents in various wildlife from Mississippi.

Because tick-borne diseases are becoming increasingly important throughout the world, monitoring their causative agents in wildlife may serve as a useful indicator of potential human exposure. We assessed the presence of known and putative zoonotic, tick-borne agents in four wildlife species in Mississippi. Animals were tested for exposure to or infection with Ehrlichia chaffeensis, Ehrlichia ewingii, Borrelia lonestari, Rickettsia spp., Anaplasma phagocytophilum, and Francisella tularensis. Whole blood and serum were tested from white-tailed deer (WTD; Odocoileus virginianus) and feral swine (Sus scrofa); serum was tested from raccoons (Procyon lotor) and opossums (Didelphis virginiana). We used polymerase chain reaction to detect all agents in blood, whereas an indirect fluorescent antibody assay was used to detect antibodies to E. chaffeensis, B. lonestari, and Rickettsia parkeri (spotted fever group rickettsiae) antigens in serum. Molecular evidence of infection with E. chaffeensis, B. lonestari, and An. phagocytophilum was detected only in WTD. Antibodies to E. chaffeensis antigen were detected in 43.9% of WTD, 32.8% of swine, 42.1% of raccoons, and 15.8% of opossums. Serologic evidence of exposure to B. lonestari antigen was found in 19.3% of WTD, 6.9% of swine, and 5.3% of raccoons, but not in opossums. Interestingly, the percent of animals with antibodies reactive to spotted fever group rickettsiae (R. parkeri antigen) was highest in raccoons (73.7%) and opossums (57.9%). These results support the role of WTD as reservoirs for E. chaffeensis, B. lonestari, and An. phagocytophilum, as well as provide additional evidence for exposure of raccoons and opossums to E. chaffeensis. Finally, we provide new data that feral swine may have antibodies to these agents. Thus, in general, these four wildlife species are exposed to tick-borne disease agents in Mississippi, suggesting that ticks carry and have the potential to transmit the agents to humans in the state.

Alterations in membrane-associated CD14 expression and the simultaneous liberation of soluble CD14 fragment in adherent macrophages mediated by a leukocyte carboxyl/aspartate protease.

Investigations sought to discover the biochemical mechanisms in macrophages that mediate the 'shedding' of soluble CD14 fragment. Stimulated macrophages display both increased liberation of soluble CD14 fragment and decreases in residual membrane-associated CD14 complexes following exposure to activating agents (fMLP/A23187). Application of 'class-specific' protease inhibitors revealed that a thiol/cysteine was involved in the biochemical production of soluble CD14 fractions and that a metalloprotease enzymatically degraded soluble CD14 fragment. Exposure of macrophages to individual proteases revealed that both cathepsin-D and elastase promoted variable depletion of membrane-associated CD14 complexes. Additionally, cathepsin-D, and to a lesser extent elastase, generated soluble CD14 fragment. Related studies isolated a carboxyl/aspartate protease from activated macrophages using pepstatin-A affinity chromatography. The physical and functional properties of macrophage pepstatin-A binding protein fractions closely corresponded with the known characteristics of cathepsin-D with respect to: (i) cellular origin; (ii) binding-avidity of carboxyl/aspartate proteases for pepstatin-A; (iii) non-specific proteolysis of haemoglobin detected by Hb-PAGE zymography; and (iv) hydrolysis of a synthetic cathepsin-D-specific peptide substrate. Interpretation of these findings collectively implies that activated leukocytes can biochemically alter membrane-associated CD14 complex expression and promote the liberation of soluble CD14 fragment in both activated and non-activated cell populations.