Entomoplasma freundtii sp. nov., a new species from a green tiger beetle (Coleoptera: Cicindelidae).

A mollicute (strain BARC 318T) isolated from gut tissue of a green tiger beetle (Coleoptera: Cicindelidae) was found by dark-field microscopy to consist of non-helical, non-motile, pleomorphic coccoid forms of various sizes. In ultrastructural studies, individual cells varied in diameter from 300 to 1200 nm, were surrounded by a cytoplasmic membrane and showed no evidence of cell wall. The organisms were readily filterable through membrane filters with mean pore diameters of 450 and 300 nm, with unusually large numbers of organisms filterable through 200 nm pore membrane filters. Growth occurred over a temperature range of 15-32 degrees C with optimum growth at 30 degrees C. The organism fermented glucose and hydrolysed arginine but did not hydrolyse urea. Strain BARC 318T was insensitive to 500 U penicillin ml-1 and required serum or cholesterol for growth. It was serologically distinct from all currently described sterol-requiring, fermentative Mycoplasma species and from 12 non-sterol-requiring Mesoplasma species, 13 non-sterol-requiring Acholeplasma species and 5 previously described sterol-requiring Entomoplasma species. Strain BARC 318T was shown to have a G + C content of 34 mol% and a genome size of 870 kbp. The 16S rDNA sequence of strain BARC 318T was compared to 16S rDNA sequences of several other Entomoplasma species and to other representative species of the genera Spiroplasma and Mycoplasma, and to other members of the class Mollicutes. These comparisons indicated that strain BARC 318T had close phylogenetic relationships to other Entomoplasma species. On the basis of these findings and other similarities in morphology, growth and temperature requirements and genomic features, the organism was assigned to the genus Entomoplasma. Strain BARC 318T (ATCC 51999T) is designated the type strain of Entomoplasma freundtii sp. nov.

Mouse interstitial lung disease and pleuritis induction by human Mollicute-like organisms.

Mollicute-Like Organisms (MLO) are cell-wall deficient intracellular bacterial pathogens. As MLO are non-cultivable, detection is based on finding typical Mollicute bodies within the host cell using a transmission electron microscope. Extracellular Mollicutes cause disease by a variety of mechanisms. MLO cause disease by similar mechanisms, and in addition directly alter the host cell nucleus, replace the cytoplasm, and destroy the organelles. MLO parasitization of plant cells causes a well studied chronic vascular disease reversible by tetracycline antibiotics. Recently similar MLO were reported to cause human chronic ocular vasculitis. As it parasitizes, lyses, and destroys leucocytes, it has been termed Leucocytoclastic MLO. Inoculation of this MLO into mouse eyelids produced delayed onset chronic ocular and lethal cardiac vasculitis. All lesions demonstrated tissue lysis with leucocytic infiltrates and MLO parasitized leucocytes. MLO-caused human and mouse disease responds to Rifampin. This report describes the 40 interstitial lung disease lesions in 21 of 100 of those MLO inoculated mice vs 0 in 200 controls (P less than 0.05) and 27 pleuritis lesions in 17 mice vs 0 control mice (P less than 0.05). The lung and pleural disease were associated in 13 lesions and unassociated in 41 lesions. MLO parasitized leucocytes were found in both the lung and pleural lesions from six of six MLO inoculated mice versus none of six controls. As most human interstitial lung and pleural diseases are idiopathic and closely resemble this mouse disease, they may be induced by MLO and treatable by Rifampin.

Mouse lethal cardiovascular disease: induction by human leucocyte intracellular Mollicutes.

Plant pathologists have known for several years that intracellular Mollicutes (M), i.e. cell wall deficient bacteria, are plant vascular pathogens, but because those M are non-cultivatable, they can only be studied by Transmission Electron Microscopy (TEM). Only recently have similar M been shown to be human and animal pathogens. Those human ocular Vasculitis (V) and mouse chronic ocular and lethal systemic V producing M parasitize vitreous polymorphonuclear leucocytes, lymphocytes, and monocytes as 'viral-like' 0.005-0.010 micron elemental particles which grow within the leucocyte into 0.01-0.03 micron diameter tubules, 0.3-1.5 micron spherules, and distinctive 0.5-0.7 micron cocci with spore-like cell walls. This report describes the 48 arteriolar and capillary sized V, Aschoff nodules, valvulitis, and myocytolytic lesions in the heart and great vessels in 18 of 100 human vitreous VM containing eyelid inoculated mice versus 0 of 200 controls (P less than 0.05) plus VM within parasitized leucocytes in 15 of 15 of those lesions by TEM. The results indicate dissemination of VM from the eyelid to produce a significant incidence of distinctive multifocal VM directly induced cardiovascular micro-V lesions that probably contributed to their excessive mortality. Because several human idiopathic diseases develop similar cardiovascular lesions a TEM search for VM parasitized leucocytes in those human diseases seems justified.

Enzymatic activities in cell fractions of mycoplasmalike organisms purified from aster yellows-infected plants.

Mycoplasmalike organisms (MLOs), purified from aster yellows-infected plants were osmotically lysed, and the membranes were separated from the cytoplasmic fraction through differential centrifugation. Electron microscopic examinations of sections of the purified MLOs and the isolated membranes showed pleomorphic bodies and unit membranous empty vesicles, respectively. Cell fractions were tested for NADH oxidase, NADPH oxidase, ATPase, RNase, DNase, and p-nitrophenyl phosphatase activity. NADH oxidase and ATPase were confined to the membrane fraction and NADPH oxidase to the cytoplasmic fraction of the MLOs. para-Nitrophenyl phosphatase, RNase, and DNase activities were detected in both membrane and cytoplasmic fractions, but p-nitrophenyl phosphatase and RNase appeared to be associated with membranes and DNase with the cytoplasmic fraction. Glucose-6-phosphate dehydrogenase was found in the cytoplasmic fraction of the MLO cells. Our findings on the distribution of enzymes in MLO cells and cell fractions are the first basic documentation on nonhelical, nonculturable microbes parasitic to plants.

Early phases of degeneration of mycoplasma-like bodies in plant tissue cultures infected with aster yellows: morphological analogies with Mycoplasma hominis.

Several forms of mycoplasma-like bodies (MLBs) were observed in electron micrographs of phloem cells from explants infected with aster yellows disease. The structures were found only in explants 11--25 days old, when the degenerating MLBs were first observed. They resembled those described by Anderson and Barile in 1965 for Mycoplasma hominis. A sequence of events in the degeneration of MLBs is postulated and morphological analogies with degenerating M. hominis are discussed.

Scanning electron microscopy of Thermoplasma acidophilum.

The scanning electron microscope was utilized to observe the morphology of the thermophilic, acidophilic mycoplasma, Thermoplasma acidophilum. Upon examination of the surface morphology, the size and shape of this unusual mycoplasma revealed its similarity to the other mycoplasmas that have been investigated.