ABSTRACT
Piscirickettsia salmonis is the etiological agent of salmonid rickettsial septicemia, an economically significant disease affecting the salmon aquaculture industry. As with other rickettsial pathogens, antigenic analysis of P. salmonis has been limited by the inherent difficulties of purifying an intracellular organism away from host cell material. In this report, we describe the use of diatrizoate meglumine and diatrizoate sodium (DMDS) density gradient centrifugation to purify P. salmonis grown in chinook salmonis was consistently concentrated in a visible band within the DMDS density gradient at density of 1.15 to 1.16 g ml(-1). Recovery of purified, viable organisms from DMDS density gradients varied from 0.6 to 3%. Preparations of uninfected CHSE-214 cells, CHSE-214 cells infected with P. salmonis, and gradient-purified P. salmonis were compared using sodium dodecyl sulfate polyacrylamide gel electrophoresis to assess the degree of purification and to identify P. salmonis-specific proteins. Although gradient-purified P. salmonis preparations were not completely free of host cell material, 8 bacterial proteins were identified. Polyclonal rabbit antiserum was used in an immunoblot of proteins from purified P. salmonis to identify 3 major and 5 minor antigens. The major antigens of 56, 30 and 20 kDa were potential candidates for experimental vaccines and development of novel diagnostic assays.
Subject(s)
Antigens, Bacterial/analysis , Fish Diseases/microbiology , Rickettsiaceae Infections/veterinary , Rickettsiaceae/isolation & purification , Salmon , Animals , Antigens, Bacterial/chemistry , Aquaculture , Bacterial Proteins/analysis , Bacterial Proteins/chemistry , Bacterial Proteins/immunology , Centrifugation, Density Gradient , Electrophoresis, Polyacrylamide Gel , Immunoblotting , Molecular Weight , Rickettsiaceae/growth & development , Rickettsiaceae/immunology , Rickettsiaceae Infections/microbiologyABSTRACT
Stainless steel wire 0.036 inch in diameter was compared with Kevlar 49 polyaramid fiber as a means of reinforcing a four-unit posterior provisional fixed restoration with 2 pontics. Three reinforcement patterns for wire and two for Kevlar 49 were evaluated and compared with the control, which was an unreinforced provisional restoration. A central tensile load was placed on the cemented provisional restoration and the variables were measured: (1) the initial stiffness; (2) the load at initial fracture; and (3) the unit toughness, or the energy stored in the beam at a point where the load had undergone a 1.0-mm deflection. Statistical analysis showed (1) the bent wire configuration had a significantly higher initial stiffness (P < or = .05), (2) there was no difference between designs for load at initial fracture, and (3) the bent wire had a significantly higher unit toughness value (P < or = .05).
