STAINING OF MICROSPORIDIAN SPORES BY OPTICAL BRIGHTENERS WITH REMARKSON THE USE OF BRIGHTENERS FOR THE DIAGNOSIS OF AIDS-ASSOCIATED HUMAN MICROSPORIDIOSES

Conditions for the effective fluorescence labelling of microsporidian spores by optical brighteners, based on the presence of chitin in the spore wall, are described. Spores of Vairimorpha ephestiae, V. necatri x, V plodiae, Nosema bombycis, N. apis, N. algerae, Encephalitozoon cu niculi and Enterocytozoon bieneusi were examined. The degree of bindin g of Calcofluor White M2R (CFW) to untreated spores depends on the con ditions and time of storage and the degree of bacterial contamination of the spore sample. Unpurified spores, stored in water are unreliable as control material for the estimation of CFW labelling. However, spo res subjected to alkaline treatment by NaOH before CFW application are visualized with ease under all experimental conditions by their brigh t, not quenching fluorescence in shortwave light (almost-equal-to 350 nm) in CFW dilutions of 10(-4) or even lower. Similiar improvement in labelling is achieved by exposing spores to CFW dissolved in 0.5-1 N N aOH. As well as Calcofluor White M2R other optical brigteners (e.g. Uv itex 2B, Ciba-Geigy or Rylux BA, Ostacolor) can be used for labelling of spores.