Description of Corynebacterium tuberculostearicum sp. nov., a leprosy-derived Corynebacterium.

Leprosy-derived corynebacteria (LDC) have been extensively studied over the past decade. A composite of their biological properties (cell morphology, staining reactions, cellular inclusions and guanine-plus-cytosine content of their deoxyribonucleic acid; 16 strains studied) and their chemical structures (peptidoglycan type, major cell wall polysaccharide, major glycolipid as well as characteristic mycolic acids) appears to define them as members of the genus Corynebacterium. In relation to other corynebacteria found in humans, including "JK corynebacteria", they seem to be distinct. They are here named Corynebacterium tuberculostearicum sp. nov. because they produce a 10-methyloctadecanoic (tuberculostearic) acid (8 strains studied). This and some of their other attributes are considered in relation to properties of leprosy bacilli and Mycobacterium leprae.

Propionibacterium, Corynebacterium, Mycobacterium and Lepra bacilli.

Evidence is presented which suggests that certain key markers of lepra bacilli reside collectively in Proprionibacterium acnes, Corynebacterium tuberculostearicum and Mycobacterium leprae. The unrestricted replication of Mycobacterium leprae depends most probably upon the presence of an immune-deficiency-inducing viral agent or possibly on the combined effects of the organisms considered.

DNA isolated from Mycobacterium leprae: genome size, base ratio, and homology with other related bacteria as determined by optical DNA-DNA reassociation.

DNA derived from Mycobacterium leprae (grown in armadillos) was isolated, purified, and analyzed spectrophotometrically. The genome size and guanine-plus-cytosine content of M. leprae were 1.3 X 10(9) and 55.8%, respectively. Among selected strains of mycobacterial, nocardial and corynebacterial species, Corynebacterium sp. 2628 LB, isolated from a human leprosy patient, showed the highest DNA homology with M. leprae. Of the DNAs derived from mycobacteria, those of M. tuberculosis and M. scrofulaceum showed a comparatively high reassociation with the DNA of M. leprae.

DNA isolated from Mycobacterium leprae: genome size, base ratio, and homology with other related bacteria as determined by optical DNA-DNA reassociation.

DNA derived from Mycobacterium leprae (grown in armadillos) was isolated, purified, and analyzed spectrophotometrically. The genome size and the guanine-plus-cytosine content of M. leprae were 1.3 x 10(9) and 55.8%, respectively. Among selected strains of mycobacterial, nocardial, and corynebacterial species, Corynebacterium sp. 2628 LB, isolated from a human leprosy patient, showed the highest DNA homology with M. leprae. Of the DNAs derived from mycobacteria, those of M. tuberculosis and M. scrofulaceum showed a comparatively high reassociation with the DNMA of M. liprae.

Chemical characterization of organisms isolated from leprosy patients.

CHEMICAL ANALYSES OF THE CELL WALLS OF ORGANISMS ISOLATED IN VARIOUS PARTS OF THE WORLD FROM CASES OF LEPROMATOUS AND TUBERCULOID LEPROSY MAKE POSSIBLE THEIR ASSIGNMENT TO ONE OF THE THREE GENERA: Corynebacterium, Mycobacterium, or Propionibacterium. One, bacterium 22M, remains unassigned. The combined chemical and enzymatic properties attributed to leprosy bacilli freshly harvested from lepromata are found collectively, but not individually, in these three genera.

Cytochemical reactions of human leprosy bacilli and mycobacteria: ultrastructural implications.

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.

Elimination of the acid fastness but not the gram positivity of leprosy bacilli after extraction with pyridine.

Bacilli from a number of tissues excised from cases of lepromatous and borderline leprosy, unlike mycobacteria, were found to lose their acid-fastness after treatment with pyridine.

Phenoloxidase activity in organisms isolated from lepromatous and tuberculoid leprosy.

Anaerobic corynebacteria (propionibacteria), isolated in pure culture from the plasma of a case of tuberculoid leprosy and from lepromata of cases of lepromatous leprosy, exhibited phenoloxidase activity such as that shown by Prabhakaran to be associated with leprosy bacilli harvested from patients suffering from leprosy. Several corynebacteria, mycobacteria, and nocardias similarly examined did not produce phenoloxidase.

Ultrastructural changes in bacteria isolated from cases of leprosy.

One of two groups of rod-shaped bacteria (bacilli) isolated from cases of human leprosy have been found, under certain cultural conditions, to give rise to unusual phenotypes. In electron micrographs of ultrathin sections and negatively stained whole cells of these osmotically fragile bacteria, ultrastructural anomalies are apparent and seem to arise from disorder in the process(es) of septum formation or cell envelope biosynthesis or both. Two of the four strains examined are related to Corynebacterium acnes.

Crystalline inclusions of bacterium 22M.

Crystalline inclusions occur in bacterium 22M. Arrays consist of parallel rows of polyhedral subunits (7.5 nm in diameter), 2.0 nm apart, with a distance of 3.0 nm between rows.