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[Infectious distribution and resistant of Neisseria gonorrhoeae, Mycoplasma, and Chlamydia trachomatis in the chronic prostatitis].

OBJECTIVE: To investigate the infectious distribution and resistant of Neisseria gonorrhoeae, Mycoplasma, and Chlamydia trachomatis in the chronic prostatitis. METHODS: The identification and susceptibility of Neisseria gonorrhoeaes and Mycoplasmas were detected by a cultural method. The nitrocefin test was used to detect the beta-lactamase in Neisseria gonorrhoeae strains. Chlamydia trachomatis was identificated by a monoclonal gold labeled antibody method. RESULTS: A total of 2,900 prostatic fluids were detected and the rates of isolation of Neisseria gonorrhoeae, Mycoplasma, and Chlamydia trachomatis were 3.3%, 12.8%, and 0.9%, respectively. The prevalence of beta-lactamase in Neisseria gonorrhoeae strains was 12.6%. The resistant percentages of Neisseria gonorrhoeae strains were 73.7% to penicillin and 91.6% to ciprofloxacin and ofloxacin respectively, but the susceptibility to spectinomycin, cephalosporin and cefoxitin was good. The resistant rates of Mycoplasma to tetracycline, acetylspiramycin, erythromycin, and ofloxacin were 50.0% or more, but the susceptibilities to roxithromycin, doxycyclin, levofloxacin, minocyclin, josamycin, and azithromycin were about 70.0%-80.0%. CONCLUSION: The isolation rate of Mycoplasma is higher than that of Neisseria gonorrhoeae and Chlamydia trachomatis in the chronic prostatitis. It is important to detect the susceptibility of Neisseria gonorrhoeae and Mycoplasma for the use of antibiotics in reason.

Adult↗

[Determination of systemic infections due to Mycoplasma in patients with clinically defined amyotrophic lateral sclerosis].

INTRODUCTION: Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease of unknown origin which has been linked to chronic infections due to intracellular microorganisms. AIMS: The purpose of this study was to identify species of Mycoplasma in blood samples from patients with clinically defined ALS by means of the polymerase chain reaction (PCR) method in comparison to healthy control subjects. PATIENTS AND METHODS: We conducted a case-control study involving 75 participants, 20 of whom were patients with clinically defined ALS and 55 healthy controls. Venous blood samples were taken and processed in the Neuroimmunology Laboratory, where they were submitted to the PCR test for Mycoplasma sp. RESULTS: The patients with ALS were between 35 and 82 years old (mean: 52.5); the ages of the healthy control subjects ranged from 35 to 60 years (mean: 44.1). After performing the PCR for Mycoplasma sp. the following results were obtained: among the patients with ALS, 10 were found to be positive (50%) and 10 were negative (50%), whereas in the control subjects we found six positives (10.91%) and 49 negatives (89.09%); these results were statistically significant (p = 0.001). On calculating the estimated risk, an odds ratio of 8167 (CI 95%: 2.4-27.6) was obtained. This indicates that the risk of suffering from ALS, if the PCR test for Mycoplasma sp. is positive, is 8:1. CONCLUSIONS: There is a strong link between suffering from a chronic infection due to Mycoplasma and developing ALS. Intracellular pathogenic agents such as Mycoplasma can play a role in the genesis of neurodegenerative diseases.

Adult↗

Mycoplasmas and acholeplasmas isolated from ducks and their possible association with pasteurellas.

Two hundred and sixty-three cases of clinically diseased ducks of all ages were examined for the presence of mycoplasmas. Mycoplasmas and acholeplasmas belonging to more than eight serogroups were cultured from 68 of them, and comprised 12 M anatis, one M columbinasale, two M gallinaceum, two M gallinarum, nine M synoviae, three unidentified Mycoplasma species, 37 Acholeplasma laidlawii and one unclassified acholeplasma belonging to each of serogroups 7 and 8. They were identified by biochemical characterisation, disc growth inhibition and agar gel diffusion tests. Fifty-three (78 per cent) of the isolates occurred with species of Pasteurella: 33.8 per cent with Pasteurella anatipestifer, 32.4 per cent with P multocida and 11.8 per cent with both P anatipestifer and P multocida. Nine of the isolates (13.2 per cent) were in pure culture and six (8.8 per cent) with other agents. Of the ducks negative for mycoplasmas 33.3 per cent were infected with P anatipestifer, 25.1 per cent with P multocida and 14.4 per cent with both P anatipestifer and P multocida. There was no correlation between the infections with mycoplasmas and P anatipestifer but there was a weak association between the infections with mycoplasmas, especially M anatis and P multocida.

Acholeplasma↗

[Diagnostic experiences in the routine restrained inspection of turkey stock for mycoplasma infections].

A combined culturing medium has been developed and has proved to clearly increase rates of detection of avian mycoplasmas, as compared to mycoplasma culturing media previously used in the GDR. The medium can be used to culture all mycoplasma species relevant to turkey. Experimental studies have shown that pH values should be between 7.0 and 7.2 in liquid culturing media and should not exceed 7.2 in mycoplasma agar, in order to be capable of isolating, in cases of mixed infections, not only Mycoplasma (M.) meleagridis but also M. gallisepticum. Culturing media should be incubated at 38.5 degrees C. The living animal should best be diagnosed by examination of palatine and cloacal swabs, with sperm being additionally checked of insemination cocks. A monitoring programme has been drafted for mycoplasma-free broods of turkey parents.

Animals↗

Enzootic pneumonia in pigs: propagation of a causative mycoplasma in cell cultures and in artificial medium.

Three strains of a new species of mycoplasma were recovered from pneumonic pig lungs, known free of Mycoplasma hyorhinis, by prolonged incubation in pig testicle cell cultures. The three strains produced a characteristic cytopathic effect in the cell cultures. A highly enriched meat-infusion-broth medium was evolved and permitted regular propagation of these organisms. Pneumonia could consistently be produced by intratracheal inoculation of pigs with the mycoplasma propagated in the enriched broth medium or in cell cultures. The mycoplasma were recovered from the lungs of experimentally infected pigs by inoculation into the broth medium. Comparative studies of the pneumonia producing mycoplasma and of M. hyorhinis were carried out in cell cultures, broth media, and in pigs infected experimentally by different routes. The morphological characteristics of the mycoplasma, grown in the different media, are described and illustrated.

Animals↗

Ultrastructural changes in hamster tracheal ring cultures exposed to Mycoplasma pneumoniae.

Ultrastructural studies of lung biopsies in this laboratory from 2 children with chronic respiratory disease and elevated titers to Mycoplasma pneumoniae showed increased pleomorphic lysosomes in alveolar macrophages. This suggested that intracellular digestion might be an important feature of mycoplasma infection and an in vitro study was undertaken. Tracheal ring cultures from 35 Syrian hamsters were exposed to Clyde M129 strain of M pneumoniae. Controls consisted of untreated tracheal rings from the same animals. Both groups were subjected to light, fluorescent, and electron microscopy. Cultures exposed to virulent mycoplasma exhibited specific immunofluorescence at mucosal surfaces and in the submucosa. Mucosal cytopathic changes and submucosal necrosis were prominent. Submucosal macrophages contained increased numbers of lysosomes, some of which contained mycoplasma-like forms. If Mycoplasma pneumoniae are ingested by macrophages, this would provide a mechanism through which immune phenomena could occur and would partially explain the prominent peribronchial lymphocytic accumulation of human mycoplasma infection.

Animals↗

[State of cellular immunity and phagocytes in guinea pigs infected with Mycoplasma pneumoniae].

The work presents the results of the study of experimental Mycoplasma pneumoniae infection in guinea-pigs. A considerable increase in the engulfment of mycoplasmas and blood leukocytes was found to occur on days 14-28 after the infection. The correlation between the degree of the engulfment of mycoplasmas by macrophages and the activity of lymphocytes in the reaction of blast-cell transformation with phytohemagglutinin and Mycoplasma antigen was observed. The peculiarities of the course of infection in sensitized animals were revealed. Virulent Mycoplasma pneumoniae strains were found to have a toxic effect on the lymphocytes and macrophages of guinea-pigs. This effect was neutralized by specific antiserum. The importance of these facts for the pathogenesis of Mycoplasma infection is discussed.

Animals↗

A survey of Mycoplasma infections in domestic poultry.

A survey of mycoplasma infections of chickens, turkeys and ducks was made on tissues from a variety of sources and birds of various ages, and from pipped and dead-in-shell turkey embryos. The tissues examined consisted mainly of respiratory tissue and the cloaca and contents and also a small number of joint lesions and other tissues. From chickens, mycoplasmas were isolated from a total of 138 tissues with Mycoplasma gallisepticum in preponderance. This was followed by M gallinarum, untyped organisms, M synoviae and a number of other mycoplasmas and Acholeplasma laidlawii. From turkeys, poults and embryos, mycoplasmas were recovered from 164 tissues with M meleagridis in preponderance. This was followed by M gallisepticum, serovar I, M synoviae and a number of other species including untyped isolates. From ducks, M gallisepticum and M anatis were recovered in equal numbers. Mycoplasma infections with more than one species occurred in the same tissue in all species of stock but especially in turkeys.

Acholeplasma↗

Pathology of goats and sheep experimentally infected with Mycoplasma mycoides var. capri.

Twenty goats and four sheep were experimentally infected with Mycoplasma mycoides var. capri and examined histopathologically. Death occurred or a moribund state developed in most of them within a week after parenteral inoculation with mycoplasmas. Histopathological examination revealed severe serofibrinous inflammation on the serosa and interstitial connective tissue at the site of inoculation. Marked inflammatory edema with infiltrating polymorphs was found in the early stage of infection. As the infection progressed, the lesion became a cellulitis; necrosis, sometimes with focal liquefaction, was a constant feature. Organization with infiltration by lymphocytes and histiocytes had begun in some tissues. Acutely infected organs had evidence of systemic involvement. Lesions included splenitis, lymphadenitis, meningitis, metastatic foci in the intestine and urinary bladder, and parenchymatous degeneration in the liver, heart, and kidneys. Mycoplasmas were isolated from many tissues of critically ill animals. Other goats which had received mycoplasmas by instillation escaped severe illness. Even so, mycoplasmas were isolated from one or two of their tissues at necropsy. The great variation in clinical and pathological response was apparently dependent upon the route of inoculation. Recovery of mycoplasmas from two contact control goats which remained healthy is suggestive of the clinically undectable carrier state. As other workers have reported, the clinical and pathological changes in sheep were similar to those in parenterally inoculated goats.

Animals↗

Intestinal mycoplasma in Crohn's disease.

Intestinal diversion with reconnection in active Crohn's disease (CD) indicates that luminal contents or bacteria contribute to the formation of CD lesions. Fluorescent staining for mycoplasma in freshly resected Crohn's tissue and electron microscopy reveal intracellular organisms akin to mycoplasma. Historically, tissue culture of CD has shown mycoplasma described as contaminants. Mycoplasma are surface epithelial parasites requiring exogenous cholesterol for membrane stability and cell entry. PCR of intestinal tissue has shown Mycoplasma pneumoniae to be detectable more significantly in CD. Oral M. iowae in experimental poultry localizes to the distal small bowel and colon. Hypothetically, lipopeptides of mycoplasmal membranes are proposed to cause chronicity and stronger immune responses than by other bacteria. 'Intestinal' mycoplasmas, from a number of observations, deserve consideration as organisms mediating inflammation of acute and chronic CD.

Animals↗

Differentiation in the murine B cell lymphoma I.29: inductive capacities of lipopolysaccharide and Mycoplasma fermentans products.

Cells from the murine B lymphoma I.29, expressing IgM or IgA of identical idiotype, were found inducible by lipopolysaccharide to differentiate into plasma cells. Within 3 days, differentiating cells lost membrane-bound immunoglobulin (Ig) and accumulated large quantities of intracytoplasmic Ig. At day 6 of culture, IgA secretion increased 50-100-fold, as determined by enzyme-linked immunoassay. Proliferation increased for the first days of culture but decreased thereafter; by day 10 very few viable cells were present in lipopolysaccharide-stimulated cultures. Similar results were obtained by culturing I.29 cells in the presence of supernatants of certain B cell lines (e.g. BFO.3). The finding of a strict correlation between the inductive activity and presence of contaminating Mycoplasma fermentans suggested that factor(s) released by mycoplasma were responsible for the mitogenic activities. This was further indicated by the findings that: the supernatants of BFO.3 that were rendered free of mycoplasma were not inductive, and a nonactive cell line could be made active by infection with supernatants of BFO.3 cells containing viable microorganisms. Thus, supernatants of mycoplasma-infected cell lines may act as potent polyclonal activators on both normal and malignant B lymphocytes. The ability to induce membrane Ig on 70Z/3 cells indicates that mycoplasma-related mitogens are also active on pre-B cells. The possibility of mycoplasma contamination should thus be carefully excluded when presumptive factors of cloned cell lines are being evaluated.

Animals↗

Mycoplasma stimulates the production of oxidative radicals by murine peritoneal macrophages.

Mycoplasmas and mycoplasma membranes have been shown to induce the production of inflammatory cytokines, including tumor necrosis factor alpha and interleukin-6, as well as nitric oxide, by mouse macrophages and rat brain astrocytes. Luminol-enhanced chemiluminescence was used as a sensitive method to show that Mycoplasma capricolum membranes induce mouse peritoneal macrophages to produce reactive oxygen radicals. Coincubation of the mycoplasma with a secondary stimulus, namely macrophage-activating factor or interferon-gamma, increased the chemiluminescence. The augmentation was abolished by the nitric oxide synthase inhibitor NG-methyl-L-arginine, indicating the involvement of nitric oxide. The coproduction of superoxide and nitric oxide by the same cell allows the formation of the powerful oxidant peroxynitrite, which could be responsible for the increased chemiluminescence. Induction of oxidizing radicals by mycoplasmas may contribute to the clinical pathology seen in mycoplasma infections.

Animals↗

Unique dicistronic operon (ptsI-crr) in Mycoplasma capricolum encoding enzyme I and the glucose-specific enzyme IIA of the phosphoenolpyruvate:sugar phosphotransferase system: cloning, sequencing, promoter analysis, and protein characterization.

The region of the genome of Mycoplasma capricolum encompassing the genes for Enzymes I and IIAglc of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) was cloned and sequenced. Examination of the sequence revealed a unique arrangement of the pts operon. In all other bacterial species characterized thus far, the gene encoding Enzyme I (ptsI) in the pts operon is located immediately downstream of the gene (ptsH) encoding HPr, a general energy coupling protein of the PTS. In M. capricolum, ptsH and ptsI reside on 2 distinct operons at separate loci on the chromosome (Zhu PP, Reizer J, Reizer A, Peterkofsky A, 1993, J Biol Chem 268:26531-26540). In the present work, it is shown that the Mycoplasma Enzyme I gene is preceded by an open reading frame homologous to the product of the Escherichia coli kdtB gene and is followed by the gene (crr) encoding Enzyme IIAglc. Northern blot analysis indicated that ptsI and crr constitute a dicistronic operon that includes an independent promoter for the crr gene. Primer extension studies established the transcription start sites for the ptsI and crr genes. The products of the ptsI and crr genes are homologous to previously sequenced Enzymes I and IIAglc proteins but are more similar to the counterpart proteins from gram-positive than to those from gram-negative organisms. The deduced amino acid sequence of the Mycoplasma Enzyme I shows that it differs from other Enzymes I by having fewer acidic amino acids and more basic, amidated, and aromatic amino acids. The deduced amino acid sequence of the Mycoplasma Enzyme IIAglc indicates that it is the shortest (154 residues) of the proteins in this class and it is the only Enzyme IIAglc with a tryptophan and a cysteine residue. In vitro sugar phosphorylation studies with extracts from E. coli and Bacillus subtilis and purified proteins indicated that the Mycoplasma HPr is not a phosphoacceptor from the E. coli Enzyme I, whereas the Mycoplasma Enzyme IIAglc accepts and transfers phosphate from both E. coli and B. subtilis PTS components.

Amino Acid Sequence↗

Detection of Mycoplasma in avian live virus vaccines by polymerase chain reaction.

The polymerase chain reaction (PCR) was evaluated to detect mycoplasma contamination of avian live virus vaccines. The specificity of the primers showed that 34 strains belonging to nine species of avian mycoplasma DNA could be detected. The sensitivity of PCR to detect mycoplasma DNA was 10(0.2) colony forming units (cfu) of Mycoplasma synoviae and 10(0.7) cfu of Mycoplasma gallisepticum. When M. synoviae and M. gallisepticum were spiked into several avian live virus vaccines, PCR gave a positive reaction except for the avian pox and the avian encephalomyelitis vaccines which were prepared from organ homogenates. Short-term incubation of avian encephalomyelitis vaccines improved the sensitivity of PCR to detect both M. synoviae and M. gallisepticum. Therefore, PCR, combined with the short-term incubation, were shown to be most effective in detecting mycoplasma contamination in all of avian live virus vaccines.

Animals↗

Physical studies of lipid organization and dynamics in mycoplasma membranes.

It should be clear from this summary that we currently know a great deal about the organization and dynamics of the lipids in mycoplasma membranes in general, and in the cell membrane of A. laidlawii in particular. In fact, research on mycoplasma membranes has been important in unambiguously establishing the fundamental lipid bilayer structure of all biological membranes and in elucidating some of the major properties of bilayers in biomembranes, such as their thermotropic phase behavior and interactions with cholesterol and membrane proteins. Although a great deal has been learned, a number of issues have not been fully resolved. In particular, the concept of membrane lipid fluidity must be refined and quantitated, and the relationship between orientational order and rates of motion better understood. This will require that the apparent discrepancies between some of the results obtained, for example, by the various spectroscopic techniques, be resolved. In particular, the nature of the boundary lipid surrounding integral membrane proteins will require further study, as will the question of the specificity of lipid-protein interactions. Also, accurate quantitative measurements for the lateral and rotational mobilities of the various lipid components in the mycoplasma membranes have not yet been made. Although not reviewed in this chapter, the related questions of the in vivo rate of phospholipid, glycolipid, and cholesterol transverse diffusion (flip-flop), and the possible asymmetric transbilayer distribution of these components in mycoplasma membranes, are still not well understood. Although much remains to be done, particularly with respect to our understanding of protein structure and function in mycoplasma membranes, a solid basis for further advances has now been laid. The many natural advantages of mycoplasma for biochemical and biophysical investigations of membrane structure and function should continue to make these organisms very useful for membrane studies for years to come.

Cell Membrane↗

Non-occurrence of Mycoplasma genitalium in clinical specimens.

Five hundred and thirteen clinical specimens, mainly from patients with urogenital inflammations, were examined for Ureaplasma urealyticum and mycoplasmas, including cultures for Mycoplasma genitalium. The study yielded 95 isolates of Ureaplasma urealyticum, 37 isolates of Mycoplasma hominis and two isolates of Mycoplasma fermentans, but no growth of Mycoplasma genitalium was obtained. It was concluded that Mycoplasma genitalium is a relatively rare inhabitant of the human urogenital tract in Israel.

Female↗

What are mycoplasmas: the relationship of tempo and mode in bacterial evolution.

In phenotype the mycoplasmas are very different from ordinary bacteria. However, genotypically (i.e., phylogenetically) they are not. On the basis of ribosomal RNA homologies the mycoplasmas belong with the clostridia, and indeed have specific clostridial relatives. Mycoplasmas are, however, unlike almost all other bacteria in the evolutionary characteristics of their ribosomal RNAs. These RNAs contain relatively few of the highly conserved oligonucleotide sequences characteristic of normal eubacterial ribosomal RNAs. This is interpreted to be a reflection of an elevated mutation rate in mycoplasma lines of descent. A general consequence of this would be that the variation associated with a mycoplasma population is augmented both in number and kind, which in turn would lead to an unusual evolutionary course, one unique in all respects. Mycoplasmas, then, are actually tachytelic bacteria. The unusual evolutionary characteristics of their ribosomal RNAs are the imprints of their rapid evolution.

Base Composition↗

Endogenous HPRT activity in mycoplasmas isolated from cell cultures.

Five mycoplasma species most frequently isolated from cell cultures were tested for the presence of endogenous hypoxanthine phosphoribosyl-transferase (HPRT) activity. All of the five, cultured in cell-free medium, contained variable but significant levels of HPRT. Two strains of M. hyorhinis exhibited a 13-fold difference in their specific HPRT activity. When infected with any of these mycoplasma species, HPRT-deficient mouse cell mutants rapidly acquired a cell-associated HPRT activity; however, the cells remained sensitive to HAT medium and resistant to 6-thioguanine. On the other hand, normal HPRT-positive cells deliberately infected with the mycoplasmas uniformly became sensitive to HAT medium. The apparent transfer of mycoplasma-specific HPRT activity to HPRT-deficient cells may be used as a sensitive measure of cell infection by these mycoplasma strains. The HPRT activities of mycoplasmas share several common properties so that they can be distinguished easily from the mammalian HPRT isozymes. Compared to the animal cell enzymes, the mycoplasmal HPRT activities are less heat stable, more strongly inhibited by 6-thioguanine, and in general migrate more slowly in electrophoresis at a neutral pH.

Acholeplasma laidlawii↗