03069nas a2200421 4500000000100000008004100001260001300042653001000055653001100065653002100076653001400097653001500111653001200126653001200138653001700150653002300167653001100190653002100201653001200222653001100234653001800245653002500263653003100288653001800319653001400337653001300351653002400364653002600388653001200414100001500426700001600441245010000457856008200557300001200639490000800651520197400659022001402633 1973 d c1973 Mar10aAcids10aBiopsy10aCatechol Oxidase10aCell Wall10aChloroform10aEnzymes10aEthanol10aEthyl Ethers10aHistocytochemistry10aHumans10aInclusion Bodies10aleprosy10aLipids10aMycobacterium10aMycobacterium leprae10aMycobacterium lepraemurium10aPeriodic Acid10aPyridines10aSolvents10aSpecies Specificity10aStaining and Labeling10aXylenes1 aFisher C A1 aBarksdale L00aCytochemical reactions of human leprosy bacilli and mycobacteria: ultrastructural implications. uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC251709/pdf/jbacter00353-0313.pdf a1389-990 v1133 a
Leprosy bacilli harvested from freshly biopsied tissue from cases of lepromatous, borderline and histoid leprosy were, in conjunction with Mycobacterium lepraemurium and representative mycobacteria, examined cytochemically with and without their pyridine-extractable acid-fastness. Unlike the mycobacteria, unextracted leprosy bacilli failed to give a positive response to the periodic acid Schiff test or to take up Sudan black B, toluidine blue O, alkaline methylene blue or safranin O. Once their acid-fastness was removed with pyridine, leprosy bacilli were stained by all of the foregoing dyes except Sudan black B, under this condition they remained gram positive. While permanent loss of acid-fastness from leprosy bacilli always resulted in a loss of acid hematein-fixing material (Smith-Dietrich-Baker tests), the reverse was not true. Mild aqueous saponification, bromination, or sequential treatment with lipase and phospholipase D resulted in a loss of acid hematein-positivity but not acid-fastness. After pyridine extraction, bromination, or aqueous saponification, true mycobacteria lost neither their acid hematein-positivity nor their acid-fastness. The acid hematein-positive material and the acid-fastness of both leprosy bacilli and mycobacteria were lost after treatment with alkaline ethanol. These cytochemical findings are discussed in the light of what is known of the ultrastructure of leprosy bacilli and mycobacteria, and of the occurrence of a dl-3, 4-dihydroxyphenylalanine oxidase in leprosy bacilli but not in mycobacteria. An effort is made to explain the rather unique cytochemical properties of leprosy bacilli. Since pyridine-extractable acid-fastness (and acid hematein-positivity) serve to distinguish human leprosy bacilli from M. lepraemurium, one or the other, or both, are suggested as bases for differentiating these two organisms in animal experiments designed to show the in vivo propagation of human leprosy bacilli.
a0021-9193