Fig 1.
Schematic alignment of N. sennetsu and related organisms showing the restrictions sites used for the RFLP.
The unique RFLP pattern of the 16 sRNA gene target, after incubation with AluI (green), BsmFI (yellow) or StyI (red) allow the differentiation of N. sennetsu, E. chaffeensis and A. phagocytophium as well as other potentially amplified organisms. The resulting fragment sizes are as follows; N. sennetsu–AluI: 345bp (uncut); BsmF1: 180bp, 80bp, 60bp, 16bp; StyI: 215bp, 127bp; E. chaffeensis–AluI: 345bp (uncut); BsmF1: 328bp, 16bp; StyI: 215bp, 127bp; A. phagocytophium–AluI: 199bp, 145bp BsmF1: 328bp, 16bp; StyI: 345bp (uncut).
Fig 2.
Electron microscopic appearance of N. sennetsu (ATCC VR-367, Miyayama strain) in DH82 canine monocyte cultures. a)
Low power micrograph of an infected cell in which a number of bacteria can be identified in the cytoplasm (arrowheads) in addition to the nucleus (N), mitochondria (Mi) and lipid droplet (L). Note the single extracellular bacterium (arrow). Bar represents 1μm. b) Enlargement of part of the cytoplasm showing a number of gram negative bacteria (B). N–nucleus. Bar represents 200nm. c) Detail of N. sennetsu (arrow) showing the gram negative bacteria limited by two unit membranes located within a membrane bound vacuole. G—Golgi stack. Bar represents 200nm.
Table 1.
Overview of confirmed N. sennetsu patients in Laos.