ABSTRACT
A bacterial staining method using fluorescent redox dye 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) is described for quantifying actively respiring bacteria that adhere to commercially pure titanium surfaces coated with cross-linked albumin. This has not been possible to date using ordinary DNA stains such as propidium iodide (PI) or Hoechst, both of which produce a very bright background. With this technique, it was demonstrated that the cross-linked albumin inhibited the adherence of Staphylococcus aureus and Staph. epidermidis to the titanium surface.
Subject(s)
Colony Count, Microbial/methods , Staining and Labeling , Staphylococcus/isolation & purification , Tetrazolium Salts/metabolism , Titanium/chemistry , Bacterial Adhesion , Bisbenzimidazole/chemistry , Fluorescent Dyes/metabolism , Microscopy, Fluorescence , Oxygen Consumption , Propidium/chemistry , Serum Albumin, Bovine/chemistry , Staphylococcus/metabolismABSTRACT
A detailed study of the pathogenesis of herpetic eye disease in the guinea pig was undertaken to further develop this animal model. Several well-known HSV-1 strains were tested for their ability to produce disease and cause acute and latent infections of the trigeminal ganglion: McKrae, KOS, McIntyre, RE, and Shealey. Two HSV-2 strains failed to cause eye infections. The Shealey strain [HSV-1 (Sh)] produced the most severe eye infections, characterized by epithelial and stromal disease, corneal vascularization and ulcerative blepharitis. Consequently, HSV-1 (Sh) was selected as the prototype strain for this study. The frequency and severity of HSV-1 (Sh) eye disease patterns was determined by a semi-quantitative rating scale, which permitted accurate monitoring of the temporal development of the disease patterns cited above. Virus shedding from infected eyes was also quantified. All of the HSV-1 strains tested established trigeminal ganglionic latency with varying frequency, although HSV-1 (Sh) latency approached 100 percent. The kinetics of acute ganglionic infection by HSV-1 (Sh) was determined, and peak virus titers occurred on the third day after corneal inoculation. This study emphasizes the usefulness of the Guinea pig model for investigations on the pathogenesis, prevention and treatment of herpetic eye infections.
