Detection of some dengue-2 virus antigens in infected cells using immuno-microscopy.

Immunoelectron microscopy was used to detect the distribution of some dengue-2 virus proteins in infected Vero and Aedes albopictus (C6/36) cells. It was found that the envelope protein (GP 60) was located in clumps on the surface of plasma membrane, and accumulated very little in the infected cytoplasm. However no envelopment of dengue-2 virus nucleocapsids through the plasma membranes was observed. In contrast, the NS 3 (P 67) protein was distributed throughout the whole cytoplasm. No specific association of this protein with the proliferated virus-induced structures was seen. The NS 1 (GP 46) protein showed almost similar distribution as the NS 3 but its quantity appeared to be lower. The NS 3 and NS 1 distribution patterns observed supported the results of immunofluorescent staining but results on E protein were not consistent in both methods.

The influence of higher temperature on dengue-2 virus infected C6/36 mosquito cell line.

Dengue-2 virus infection of C6/36 cells was studied at 28 and 37 degrees C. In infected cells maintained at 28 degrees C, syncytial development was seen on day 4 postinfection, whereas at 37 degrees C, extensive syncytial development was seen by 32 h. Extracellular virus titre was found to correlate with the cytopathic changes. Nine Dengue-2 virus specified proteins were observed in polyacrylamide analyses of cytoplasmic extracts of C6/36 infected cells. All the proteins were observed, although in varied intensities by 32 h postinoculation at 37 degrees C and only on day 4 postinoculation at 28 degrees C. The GP60 glycoprotein appeared at 32 h postinfection when the cells were maintained at 37 degrees C and became prominent only on day 5 at 28 degrees C. The results revealed that a higher temperature accelerated the onset of cytopathic effects, hastened the development of virus specified proteins, and also enhanced the titre of extracellular infectious virus. The importance of the accumulation of the envelope protein GP60 for the development of CPE was indicated.

Prevalence of antibodies to Toxoplasma gondii in striped skunks from Saskatchewan and Alberta.

Fifteen percent (81 of 542) of striped skunks, Mephitis mephitis, collected in the prairie of Alberta and Saskatchewan during 1974 to 1978, were positive for antibodies against Toxoplasma gondii. The seropositive rates varied from 8% (6 of 78) for skunks less than six months of age to 47% (9 of 19) in animals three or more years old. Spring and summer transmission was indicated by a preponderance of high titres (greater than or equal to 1:1024) in seropositive skunks collected April through September (22 of 40, 55%) compared to seropositives collected October through MFarch (10 of 38, 26%) (P = < 0.05). Prevalence was significantly greater among skunks collected in the relatively humid parkland (63 of 286, 22%) than in the arid prairie grassland biome (20 of 225, 8%) (P = < 0.01). The results indicate that T. gondii is focally enzootic in Alberta and Saskatchewan.