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Biomedical subjects

D L Rose

Publications and source records attributed to D L Rose.

At least 19 recordsLinked to original sources

Fatal septicemia due to Mycoplasma arginini: a new human zoonosis.

A 64-year-old slaughterhouse worker with advanced non-Hodgkin's lymphoma developed septicemia and pneumonia. Mycoplasma arginini, a wall-free prokaryote found in a variety of domestic animal hosts, was repeatedly isolated from blood and bronchial washings from the patient. Immunosuppression, in part caused by hypogammaglobulinemia, probably played a key role in predisposing the patient to a fatal infection. This case suggests that animal mycoplasmas should be considered in the list of infectious agents acquired by immunosuppressed hosts.

Bacteremia

Mycoplasma penetrans sp. nov., from the urogenital tract of patients with AIDS.

An unusual mycoplasma, which was isolated from the urine of a human immunodeficiency virus-positive male homosexual patient, has an elongated flask shape and two unique sharply divided internal compartments. The tiplike compartment is densely packed with fine granules, and the body compartment is loosely filled with coarse granules consistent with ribosomal structures. The organism has properties of adherence, hemadsorption, and cytadsorption and invades many different types of mammalian cells. Adhesion and penetration apparently involve the terminally located tiplike structure. Cholesterol is required for growth, and the mycoplasma ferments glucose and hydrolyzes arginine, but does not hydrolyze urea. The results of DNA homology studies revealed that this organism is not genetically related to previously described mycoplasma species that have the same biochemical properties. The results of serologic studies demonstrated that this organism is antigenically distinct from all previously described mycoplasmas. We propose that this new mollicute species should be named Mycoplasma penetrans sp. nov. The type strain is strain GTU-54-6A1 (= ATCC 55252).

Acquired Immunodeficiency Syndrome

Identification of a plant-derived mollicute as a strain of an avian pathogen, Mycoplasma iowae, and its implications for mollicute taxonomy.

Strain PPAV, a filamentous but nonhelical mollicute, was isolated from aborted apple seeds in France in late 1979. This organism grew well in SP-4 broth, fermented glucose, and required sterol for growth, and most of its properties suggested that it belonged to the genus Mycoplasma. However, it was serologically distinct; in addition, unlike other Mycoplasma species, genome measurements consistently yielded values of about 1,000 MDa (ca. 1,500 kbp), and the organism had a growth temperature optimum of 43 degrees C. A comparison of strain PPAV 16S rRNA sequences with those of other mollicutes revealed a high degree of sequence similarity to a strain of Mycoplasma iowae, which is commonly encountered in poultry. This relationship was confirmed by performing a restriction endonuclease pattern analysis and DNA-DNA hybridization tests. The genome size of type strain 695 of M. iowae was determined to be about 1,000 MDa (1,500 kbp) by renaturation kinetics, a value which is much higher than any other value known in the genus. Additional measurements by pulsed-field gel electrophoresis yielded values of 1,300 kbp for both strain PPAV and M. iowae. Subsequent phenotypic comparisons supported this relationship. Serologic tests with strain PPAV and other strains of M. iowae confirmed the findings of other investigators that this species is serologically heterogeneous. The high optimum temperature for growth of strain PPAV was also shared by a number of M. iowae isolates. Genome size is an inappropriate character for taxonomic assignment to the family Mycoplasmataceae because strain PPAV and other established species in this family are now known to have genomes ranging in size from 1,000 to 1,400 kbp.

Animals

Mycoplasma lactucae sp. nov., a sterol-requiring mollicute from a plant surface.

Strain 831-C4T (T = type strain), isolated from the surface of lettuce plants (Lactuca sativa) obtained from a retail food market, was shown to be a sterol-requiring mollicute. Morphological examination of this organism by electron and dark-field microscopic techniques showed that it consists of small, nonhelical, nonmotile, pleomorphic coccoid cells, with individual cells surrounded by a single cytoplasmic membrane. No evidence of a cell wall was observed. The organism grew rapidly in all conventional culture medium formulations for mollicutes in either aerobic or anaerobic environments. The optimum temperature for growth was 30 degrees C, but multiplication occurred at 18 to 37 degrees C. Strain 831-C4T catabolized glucose, but hydrolysis of arginine or urea could not be demonstrated. The genome size of strain 831-C4T was determined to be about 569 megadaltons, while the base composition (guanine-plus-cytosine content) of the DNA was 30.0 mol%. Recent studies in which we compared the 16S rRNA sequences of strain 831-C4T with those of more than 40 other mollicutes indicated that this organism is phylogenetically related to the Spiroplasma-Mycoplasma mycoides clade. Strain 831-C4T was serologically unrelated to the type strains of previously described Mycoplasma species and to 18 other unclassified sterol-requiring isolates cultivated from various animal, plant, or insect sources. Strain 831-C4T (= ATCC 49193) is the type strain of Mycoplasma lactucae sp. nov.

Mycoplasma

Mycoplasma melaleucae sp. nov., a sterol-requiring mollicute from flowers of several tropical plants.

Three sterol-requiring mollicutes from floral surfaces of two tropical plant species (Melaleuca quinquenervia and Melaleuca decora) and a single isolate from a flower of the silk oak (Grevillea robusta) were serologically indistinguishable. Strain M1T (T = type strain), isolated from Melaleuca quinquenervia, was chosen for characterization. Light and electron microscopic observations of strain M1T revealed nonhelical, nonmotile, pleomorphic coccoid cells surrounded by a single cytoplasmic membrane. No evidence of a cell wall was observed. The organism grew well in SP-4 medium, but no sustained growth occurred in conventional mycoplasma media containing horse serum. The optimum temperature for growth was 23 degrees C, but multiplication occurred over a temperature range of 10 to 30 degrees C. Growth was not observed at temperatures above 30 degrees C. Strain M1T and related strains (strains M5, M10, and SO1) catabolized glucose but hydrolyzed neither arginine nor urea. The size of the strain M1T genome was about 561 megadaltons, while the guanine-plus-cytosine content of the DNA was about 27.0 mol%. The organism was serologically unrelated to the type strains of the 80 previously recognized Mycoplasma species or to 18 other unclassified sterol-requiring strains cultivated from animal, plant, or insect sources. Recent sequencing studies of 16S rRNA demonstrated that strain M1T is a member of a clade that contains the type species of the genus Mycoplasma. Strain M1 (= ATCC 49191) is the type strain of Mycoplasma melaleucae sp. nov.

Cholesterol

Mycoplasma somnilux sp. nov., Mycoplasma luminosum sp. nov., and Mycoplasma lucivorax sp. nov., new sterol-requiring mollicutes from firefly beetles (Coleoptera: Lampyridae).

Strain PYAN-1T (T = type strain), which was isolated from a pupal gut of the firefly beetle Pyractonema angulata, and strains PIMN-1T and PIPN-2T, which were isolated from guts of adult Photinus marginalis and Photinus pyralis fireflies, respectively, were demonstrated to be sterol-requiring mollicutes. Cells of the three strains were shown by electron and dark-field microscopy to be small, pleomorphic, nonhelical, nonmotile bodies surrounded by single membranes. No evidence of a cell wall was observed, and the organisms were not susceptible to 500 U of penicillin per ml. The three strains grew rapidly in SP-4 broth medium. Strains PIMN-1T and PIPN-2T grew in medium supplemented with bovine serum fraction, but strain PYAN-1T did not. All three strains grew on solid media when the cultures were incubated aerobically, but only strains PYAN-1T and PIPN-2T formed colonies when anaerobic conditions were employed. The three strains catabolized glucose but hydrolyzed neither arginine nor urea. All of the strains grew at temperatures of 18 to 32 degrees C; strains PYAN-1T and PIMN-1T also grew at 10 degrees C. The optimal temperature for growth for strains PYAN-1T and PIPN-2T was 30 degrees C; strain PIMN-1T grew equally well at 30 or 32 degrees C. None of the three strains grew at 37 degrees C. The genome sizes of strains PYAN-1T, PIMN-1T, and PIPN-2T were about 527 (478 to 589), 570 (480 to 630), and 762 (635 to 871) megadaltons, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

A phylogenetic analysis of the mycoplasmas: basis for their classification.

Small-subunit rRNA sequences were determined for almost 50 species of mycoplasmas and their walled relatives, providing the basis for a phylogenetic systematic analysis of these organisms. Five groups of mycoplasmas per se were recognized (provisional names are given): the hominis group (which included species such as Mycoplasma hominis, Mycoplasma lipophilum, Mycoplasma pulmonis, and Mycoplasma neurolyticum), the pneumoniae group (which included species such as Mycoplasma pneumoniae and Mycoplasma muris), the spiroplasma group (which included species such as Mycoplasma mycoides, Spiroplasma citri, and Spiroplasma apis), the anaeroplasma group (which encompassed the anaeroplasmas and acholeplasmas), and a group known to contain only the isolated species Asteroleplasma anaerobium. In addition to these five mycoplasma groups, a sixth group of variously named gram-positive, walled organisms (which included lactobacilli, clostridia, and other organisms) was also included in the overall phylogenetic unit. In each of these six primary groups, subgroups were readily recognized and defined. Although the phylogenetic units identified by rRNA comparisons are difficult to recognize on the basis of mutually exclusive phenotypic characters alone, phenotypic justification can be given a posteriori for a number of them.

Base Sequence

Isolation and characterization of Mycoplasma genitalium strains from the human respiratory tract.

Mycoplasma genitalium, an organism first isolated from the urethras of two men with nongonococcal urethritis, has been found in throat specimens from military recruits participating in an inactivated Mycoplasma pneumoniae vaccine field trial in 1974-1975. Four of 16 preserved throat isolates, previously identified as strains of M. pneumoniae, have now been shown to be mixtures of M. pneumoniae and M. genitalium. Purification of these mixed mycoplasmas by selection of single colonies confirmed the presence of M. genitalium. Identification of M. genitalium was based upon the occurrence of a species-specific 140-kilodalton protein adhesin in these isolates and their serologic reactivity to an M. genitalium antiserum. The frequent occurrence of both M. pneumoniae and M. genitalium in a number of these throat specimens, in combination with their shared antigenic cross-reactivities, suggests the likelihood that M. genitalium strains are easily missed in the usual laboratory identification procedures. What role M. genitalium may play in human respiratory disease remains to be determined.

Bacterial Proteins

Further characterization of an unusual plant Mollicutes species of uncertain taxonomic status.

PPAV is a type of Mollicutes that was isolated in SP4 medium from seeds of apples affected by proliferation, a disease in which mycoplasma-like organisms (MLOs) are involved. However, PPAV is probably not the etiological MLO agent for at least two reasons: 1) optimal temperature growth is 43 C, and 2) in spite of numerous isolation attempts over several years, no second PPAV culture could be obtained. PPAV is surrounded by a single cytoplasmic membrane and forms typical fried egg-shaped colonies on solid medium. The organism grows in simplified mycoplasma media, such as BSR. In growth inhibition, it shows no serological relationships with any other mycoplasmas or acholeplasmas, including those cultured from the surfaces of plants. The absence of relatedness of PPAV to other Mollicutes was confirmed by DNA hybridization studies. The genome of PPAV is close to 10(9) daltons and contains 25.2 mol % G + C. Its genome size is similar to that of Acholeplasma spp. and Spiroplasma spp. It has, however, a clearcut sterol requirement and therefore cannot be an acholeplasma. Neither is it a spiroplasma since, though filamentous in BSR medium, it has never shown signs of helicity. Hence, PPAV is a taxonomical paradox.

Acholeplasma

Characterization of some new insect-derived acholeplasmas.

Further analysis of three sterol-nonrequiring Mollicutes (strains PS-1, TAC, and YJS) isolated from gut fluids of insects confirms their similarity to Acholeplasma. They are serologically distinct from acholeplasmas of vertebrates and several other sterol nonrequiring Mollicutes isolated from plant surfaces. The PS-1 strain had a DNA G + C content of 31 mol % and a genome size of 1,030 megadaltons (MDa). Optimum temperature is in the range of 23 to 30 C. Thirty-two new nonhelical Mollicutes isolated from a much wider range of insect hosts were examined for acholeplasmas. Twenty-five of the insect isolates were grown consistently in serum-free broth, with or without Tween 80 supplements. Two of the acholeplasmas were serologically related to Acholeplasma florum, 13 strains were serologically identical to the TAC isolate reported earlier, and 10 of the putative acholeplasmas could not be identified with current reference antisera. Seven of the new nonhelical insect isolates appeared to be sterol-requiring Mollicutes. One sterol-requiring isolate (ELCN-1) was recovered from the hemolymph of a firefly, and is the first report of nonhelical Mollicutes in the insect hemocoel. Two of the seven sterol-requiring Mollicutes, which were nonhelical in earlier passages in broth, later reverted to typically helical spiroplasmas. Confirmation of sterol-requiring, nonhelical Mollicutes in insects would provide an important ecological finding that insects constitute an important reservoir for both acholeplasmas and mycoplasmas.

Acholeplasma

Urogenital challenge of primate species with Mycoplasma genitalium and characteristics of infection induced in chimpanzees.

Eighteen male and eight female primates, representing five subhuman species, were inoculated urogenitally with Mycoplasma genitalium, a microorganism recovered from men with nongonococcal urethritis. Male rhesus (Macaca mulatta) and cynomolgus (Macaca fascicularis) monkeys apparently were resistant. Female squirrel monkeys (Saimiri sciureus) and female tamarins (Saguinus mystax) exhibited low-level, genital-tract infections. Male chimpanzees (Pan troglodytes) developed an obvious genital-tract infection, with some shedding organisms for 21 weeks. M. genitalium was recovered from the blood of two of the male chimpanzees, usually when large numbers of organisms were in the urethra. Female chimpanzees generally shed organisms for 12-15 weeks. Most chimpanzees colonized with the organism exhibited increased numbers of polymorphonuclear leukocytes in the genital tract and developed a significant antibody response. The results offer substantial evidence for the pathogenicity of M. genitalium for the urogenital tract of higher primates and suggest the microorganism may have a role in human genital-tract infections.

Animals

Intraspecies genetic relatedness among strains of Acholeplasma laidlawii and of Acholeplasma axanthum by nucleic acid hybridization.

This study compares the intraspecies genetic relatedness among strains of two established species of Acholeplasma. Radiolabelled DNA probes were prepared from three strains of Acholeplasma laidlawii and two strains of Acholeplasma axanthum, by using the nick translation method. The labelled DNA probes of these two strains were hybridized to an excess of unlabelled DNA from 12 strains of Acholeplasma laidlawii and from six strains of Acholeplasma axanthum, respectively. Nucleic acid hybridization analyses showed a wide variation among strains within each of the two established species, ranging from 48 to 100% homology. The results demonstrate that strains isolated from diverse hosts and habitats within a given species of Acholeplasma exhibit extensive genotypic variations.

Acholeplasma

DNA cleavage patterns as indicators of genotypic heterogeneity among strains of Acholeplasma and Mycoplasma species.

Electrophoretic patterns of digestion products of Acholeplasma and Mycoplasma DNA by restriction endonucleases were compared. The patterns of Acholeplasma axanthum strains isolated from a variety of hosts and habitats differed markedly from each other, indicating considerable genotypic heterogeneity among strains included in this species. Heterogeneity was less marked among the Acholeplasma oculi strains tested, and was minimal among strains of the avian pathogen Mycoplasma gallisepticum. Strains of Mycoplasma genitalium isolated from the urethra of patients with non-gonococcal urethritis and from the urethra of an experimentally infected chimpanzee yielded identical cleavage patterns, indicating a high degree of genetic homogeneity of these strains. The data support the notion that mycoplasma species of strict host and tissue specificity exhibit marked genetic homogeneity. The advantages and deficiencies of the use of DNA cleavage patterns for classification purposes are discussed.

Acholeplasma

Nucleic acid relationships among Acholeplasma species.

3H-labeled Acholeplasma DNA probes were generated in vitro by the nick-translation method and used to determine the nucleotide sequence homology among the type strains of the eight currently recognized species of Acholeplasma. Very little nucleotide sequence homology (less than or equal to 18%) was found among the eight species, with heteroduplexes showing at least 12% or more mismatching as determined by thermal elution midpoints. The small amount of nucleotide sequence homology among the eight species indicates that these species are quite distinct and are not closely related to each other genomically.

Acholeplasma

Growth characteristics of large- and small-colony types of Mycoplasma mycoides subsp. mycoides on 5% sheep blood agar.

Mycoplasma mycoides subsp. mycoides of the large-colony (LC) type was isolated in pure culture on 5% sheep blood agar plates inoculated with lung specimens from a 4-month-old Toggenburg goat. The growth characteristics of this isolate, of four known LC types, and of five known small-colony (SC) types of M. mycoides subsp. mycoides were compared on 5% sheep blood agar at 2, 5, and 7 days. The SC types were not visible at 2 days and did not grow larger than 0.1 mm, whereas the LC types were visible in 2 days and increased in diameter over 7 days to between 0.4 and 0.7 mm. These results indicate that growth on 5% sheep blood agar can be used as an additional marker in differentiating LC and SC types of M. mycoides subsp. mycoides.

Agar

A newly discovered mycoplasma in the human urogenital tract.

A new mycoplasma, serologically distinct from all other known mycoplasmas, was isolated from urethral specimens from two of thirteen men with non-gonococcal urethritis. Repeatable isolation and propagation was accomplished by use of a special culture medium. The organisms adhered to glass or plastic, erythrocytes, and monkey kidney cells. This property appears to be associated with surface material restricted to the area of a terminal structure of the flask-shaped mycoplasmas. Although the data are insufficient to implicate the new mycoplasmas in human disease, the fact that they are unique, extremely fastidious, and have adherence properties, has stimulated efforts to assess their pathogenicity and possible role in human urogenital disease.

Culture Media

Helical mycoplasmas (spiroplasmas) from Ixodes ticks.

A new spiroplasma isolated from Ixodes pacificus collected in Oregon was serologically and morphologically distinct from known spiroplasmas. The new spiroplasma could also be isolated in tick cell cultures. Discovery of a new fastidious mycoplasma in ticks offers opportunities to explore the possible role of these agents in human and animal diseases.

Animals

Urogenital mycoplasma infections of man: a review with observations on a recently discovered mycoplasma.

Ureaplasma urealyticum organisms (ureaplasmas), Mycoplasma hominis, M. fermentans, M. primatum, M. Salivarium and M. pneumoniae have been isolated from the genitourinary tract. The first two of these microorganisms are found most frequently. M. hominis is a cause of some cases of postpartum and postabortal fever, acute pyelonephritis and pelvic inflammatory disease. Ureaplasmas have been associated with chorioamnionitis, habitual spontaneous abortion, low birthweight, the urethral syndrome in women, and nongonococcal urethritis (NGU) in men; but the difficulty of proving an etiological relationship is emphasized. However, in NGU there is accumulating evidence to indicate that ureaplasmas cause some cases. Some patients suffering from NGU, from whom ureaplasmas, mycoplasmas and chlamydiae cannot be isolated, respond to tetracycline therapy. This has suggested that a tetracycline-sensitive microorganism might be responsible. In this context, the isolation of a glucose-metabolizing mycoplasma from the genitourinary tracts of 2 of 13 men with NGU is of interest. This mycoplasma, serologically different from all other tested, has the structural and biological features of a pathogenic organism.

Abortion, Habitual