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PubMed2001

Gender is a major factor in determining the severity of mycoplasma respiratory disease in mice.

Gender is a significant factor in determining the susceptibility to and severity of pulmonary diseases in both humans and animals. Murine respiratory mycoplasmosis (MRM), due to Mycoplasma pulmonis infection, is an excellent animal model for evaluation of the role of various host factors on the development of acute or chronic inflammatory lung diseases. MRM has many similarities to mycoplasma respiratory disease in humans. The purpose of the present study was to determine whether gender has a significant impact on lung disease due to M. pulmonis infection in mice. It was demonstrated that male mice consistently developed more severe disease in the lung parenchyma than did female mice. There was no gender difference in disease severity along the airways or any difference in mycoplasma numbers in lungs of male and female mice. Furthermore, surgical removal of reproductive organs reduced the severity of mycoplasma disease and the numbers of mycoplasma organisms recovered from lungs. Thus, gender plays a significant role in determining the severity of M. pulmonis disease. In fact, the gender of the host was a major factor in determining whether an acute or chronic inflammatory lung disease developed after infection with M. pulmonis.

Animals↗

PubMed1963

GROWTH INHIBITORY PROPERTIES OF MYCOPLASMA ANTIBODY.

Bailey, Jack S. (The George Washington University, Washington, D.C.), Harold W. Clark, William R. Felts, and Thomas McP. Brown. Growth inhibitory properties of Mycoplasma antibody. J. Bacteriol. 86:147-150. 1963.-A substance in antiserum responsible for growth inhibition of Mycoplasma was found to be associated with the water-insoluble globulin fraction. This fraction and the agglutinins removed from agglutinated antigen inhibited the growth of Mycoplasma in a similar manner. Sonic oscillation of antiserum broth cultures partially relieved the growth inhibition. The results of this study suggest the existence of neutralized Mycoplasma in a latent phase, which could explain their infrequent detection in tissues and fluids. The application of such techniques as sonic oscillation may improve the recovery and identification of Mycoplasma from these sources.

Antibodies↗

PubMed1964

PRODUCTION AND BIOLOGICAL CHARACTERISTICS OF AN EXTRACELLULAR NEUROTOXIN FROM MYCOPLASMA NEUROLYTICUM.

Tully, Joseph G. (National Institute of Allergy and Infectious Diseases, Bethesda, Md.). Production and biological characteristics of an extracellular neurotoxin from Mycoplasma neurolyticum. J. Bacteriol. 88:381-388. 1964.-Toxigenic strains of Mycoplasma neurolyticum were found to produce a potent extracellular neurotoxin for mice under a variety of cultural conditions. The observed biological characteristics of this toxin were quite similar to those originally described by Sabin. The toxin was again found to be readily inactivated by prolonged incubation and, when separated from Mycoplasma cells by filtration, possessed a greater heat resistance than whole cells. Young mice and rats were generally susceptible to neurotoxin, whereas most other experimental animals were resistant. Filtration results confirmed a minimal reproductive unit of 125 to 200 mmu for two neurotoxic Mycoplasma strains, with both cell-free filtrates reproducing the rolling disease syndrome in mice. Twice-washed Mycoplasma cells were also observed to be as neurotoxic as filtrates. Plate counting results revealed an intravenous ld(50) of approximately 3 x 10(8) washed cells for young mice.

Animals↗

PubMed1996

Diagnosis of contagious caprine pleuropneumonia by detection and identification of Mycoplasma capricolum subsp. capripneumoniae by PCR and restriction enzyme analysis.

Contagious caprine pleuropneumonia (CCPP), one of the most serious and dramatic diseases of goats, is caused by Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae). This organism is very difficult to isolate and to correctly identify. In a previous report we described a method for the rapid detection and identification of M. capripneumoniae. This method is based on a PCR system by which a segment of the 16S rRNA gene from all mycoplasmas of the M. mycoides cluster can be amplified. The PCR product is then analyzed by restriction enzyme cleavage for the identification of M. capripneumoniae DNA. This system has now been further evaluated with respect to specificity and diagnostic efficacy for the identification and direct detection of the organism in clinical material. Identification by restriction enzyme analysis of amplified DNA from mycoplasmas of the M. mycoides cluster was verified for 55 strains, among which were 15 strains of M. capripneumoniae. The PCR was applied to clinical samples from the nose, ear, pharynx, pleural fluid, and lung tissue containing M. capripneumoniae or other mycoplasmas. As expected, mycoplasmas belonging to the M. mycoides cluster could be detected by the PCR. Restriction enzyme analysis of the PCR products could then be applied for the identification of M. capripneumoniae. Clinical samples and cultures containing M. capripneumoniae were dried on filter paper, to try an easier sample transport method, and were tested by PCR. M. capripneumoniae DNA could be detected in the dried specimens, but the sensitivity of the PCR test was reduced.

Animals↗

PubMed1997

High rates of genital mycoplasma infection in the highlands of Papua New Guinea determined both by culture and by a commercial detection kit.

Duplicate vaginal swabs were collected from 100 women, and comparisons were made between an in-house broth-agar culture system and a commercially available kit, the Mycoplasma IST kit (bioMérieux), for the detection of Mycoplasma hominis and Ureaplasma urealyticum. There was good agreement between the two systems for detection of the genital mycoplasmas in terms of sensitivity, with values of > 92% being obtained. In terms of specificity, the mutual comparisons were less favorable, though specificity values of > 72% were obtained. Statistically there was no significant difference in the performance of the two tests (P < 0.1 for both M. hominis and U. urealyticum). While the broth-agar culture system was considerably less expensive than the kit, the Mycoplasma IST kit provided additional information on antibiotic susceptibilities and had the advantages of a shelf life of up to 12 months and not requiring the preparation of culture media. The prevalences of colonization obtained for M. hominis and U. urealyticum were extremely high in this randomly selected group of women from periurban and rural settlements in the Eastern Highlands of Papua New Guinea, being > or = 70% for M. hominis and > or = 78% for U. urealyticum. colonization with both genital mycoplasmas simultaneously was also very common, with > or = 60% of women being colonized by both M. hominis and U. urealyticum.

Adolescent↗

PubMed2002

Phylogeny-based rapid identification of mycoplasmas and ureaplasmas from urethritis patients.

Some strains of mycoplasmas and ureaplasmas (family Mycoplasmataceae) are associated with nongonococcal urethritis (NGU) or other genitourinary infections. We have developed a rapid and reliable method of identifying the presence and prevalence of mycoplasmas and ureaplasmas in men with NGU. This method is based on the amplification of a part of the 16S rRNA gene by PCR and phylogenetic analysis. A portion of the 16S rRNA gene from 15 prototype strains was amplified with a set of common primers, and their nucleotides were sequenced. The nucleotide sequence of the V4 and V5 regions was analyzed by the neighbor-joining method. The 15 prototype strains were grouped into three distinct clusters, allowing us to clearly segregate the strains into distinct lineages. To determine the prevalence of these pathogens among patients with NGU, this protocol was tested with 148 urine samples. Amplifications were observed for 42 samples, and their nucleotide sequences were analyzed along with those of the 15 prototype strains. The phylogenetic tree thus constructed indicated that 15 of the 42 formed a cluster with Mycoplasma genitalium. Among the remaining specimens, 2 formed a cluster with Mycoplasma hominis, 19 with Ureaplasma urealyticum, and 5 with Ureaplasma parvum; the remaining sample contained both M. genitalium and U. urealyticum. This phylogeny-based identification of mycoplasmas and ureaplasmas provides not only a powerful tool for rapid diagnosis but also the basis for etiological studies of these pathogens.

DNA, Bacterial↗

PubMed1995

Detection by polymerase chain reaction of all common Mycoplasma in a cell culture facility.

The identification of cell cultures contaminated with organisms from the class Mollicutes has led us to examine the effectiveness of polymerase chain reaction (PCR) for detecting these organisms in genomic DNA. We developed a previously identified nested PCR primer set and compared its ability to detect Mycoplasma with that of a commercially available PCR kit for detecting Mycoplasma. We found that although the commercial system detected and identified a few of the most common Mycoplasma species, the primer set (GPO-1, GPO-2, MGSO) detected the presence of all the common Mycoplasma species and many of the rare mycoplasma species previously encountered in tissue culture.

Animals↗

PubMedDate not supplied

Efficacy of roxithromycin in the treatment of mycoplasma pneumonia.

Macrolide and tetracycline antibiotics, which are protein synthesis inhibitors, are effective in the treatment of mycoplasma pneumonia. We evaluated the clinical efficacy and safety of roxithromycin, a new macrolide antibiotic, in the treatment of mycoplasma pneumonia. Roxithromycin was administered orally to patients who had been definitely diagnosed through Mycoplasma pneumoniae isolation or serum antibody titer as having mycoplasma pneumonia. The efficacy assessment was based on clinical signs and symptoms of infection as well as bacterial culture from clinical samples. Clinical efficacy was excellent in 6 cases, good in 6 cases and fair in 1 case, with an efficacy rate of 92.3%. The bacteriological effect was evaluated in 6 patients: the organism was eradicated in 4 cases and unchanged in 2 cases. In this study, the MIC of roxithromycin against M. pneumoniae fell in the range 0.0156-0.00625 mg/l. No adverse reaction was observed. As for abnormal laboratory findings, two cases showed elevated serum glutamic-pyruvic transaminase (S-GPT), one elevated serum glutamic-oxaloacetic transaminase and S-GPT, and one reduced neutrophil counts. From our results, we consider that roxithromycin is useful in the treatment of mycoplasma pneumonia.

Administration, Oral↗

PubMed1996

Effects of interleukin-2 and cyclosporin A on pathologic features in Mycoplasma pneumonia.

To elucidate the immunopathologic mechanisms of Mycoplasma pneumonia, the effects of interleukin-2 (IL-2) and cyclosporin A (CYA) on Mycoplasma pneumonia in mice were investigated. Mice were intranasally inoculated with Mycoplasma pulmonis (M. pul) and treated with IL-2, CYA, or minocycline (MINO) every day between Days 3 and 9. They were killed at Days 7, 14, or 21 after the inoculation. Cell-mediated immunity (CMI) of the host was assessed by delayed-type hypersensitivity (DTH) to sheep red blood cells (SRBC). Peribronchial and perivascular lymphocyte cuffing and the accumulation of macrophages at the ends of bronchioles were exacerbated (p < 0.05) in IL-2-treated mice at Day 14 and reduced (p < 0.05) in CYA-treated mice at Day 7. Although the DTH responses to SRBC of saline-inoculated, IL-2-treated mice were increased at Days 7 (p < 0.01) and 14 (p < 0.05), those of M. pul-inoculated, IL-2-treated mice at Day 7 could not recover to the control level. The CMI levels of M. pul-inoculated, CYA-treated mice were decreased at Days 7, 14 (p < 0.05), and 21 (p < 0.01). Mycoplasma organisms in the lung showed the greatest decrease (p < 0.05) in MINO- and IL-2-treated mice, but increased at Day 21 (p < 0.05) in mice treated with IL-2 alone. These results suggest that the pathologic features of Mycoplasma pneumonia could be modified by the degree of host CMI.

Animals↗

PubMed1989

Short-term exposure to nitrogen dioxide enhances susceptibility to murine respiratory mycoplasmosis and decreases intrapulmonary killing of Mycoplasma pulmonis.

In C57BL/6N and C3H/HeN mice known to be free of all murine pathogens and matched for age, sex, microbiologic, and environmental factors, exposure to NO2 for 4 h prior to exposure to infectious aerosols of Mycoplasma pulmonis resulted in potentiation of murine respiratory mycoplasmosis (MRM). In the C57BL/6N mice, NO2 increased the incidence of death, incidence of gross lung lesions, and incidence of microscopic lung lesions, but did not increase the incidence of infection in the lungs. Nitrogen dioxide affected the C3H/HeN mice (a strain known to be more susceptible than the C57BL/6N strain to MRM) similarly, with the exception that the incidence of death and microscopic lesions were not affected in this strain at the concentrations of M. pulmonis used. Exposure to the oxidant also increased the severity of microscopic lesions and the numbers of Mycoplasma organisms in the lungs of both mouse strains. Thus, NO2 appeared to affect host lung defense mechanisms responsible for limiting the extent of infection. The NO2 exposure level required to produce potentiation varied with the genetic background of the host, the number of Mycoplasma organisms administered, and the end point measured. In further experiments in C57BL/6N mice, exposure to 5 or 10 ppm of NO2 for 4 h prior to infection with aerosolized, radiolabeled M. pulmonis reduced clearance of these organisms from the lungs over a 72-h time period. Nitrogen dioxide exposure did not change the rate of physical removal of Mycoplasma organisms from the lung. Reduced clearance was due to impaired intrapulmonary killing of Mycoplasma organisms in NO2-exposed mice.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Inhalation↗

PubMed2006

Molecular and antigenic characterization of a Mycoplasma bovis strain causing an outbreak of infectious keratoconjunctivitis.

An unusually high incidence of infectious keratoconjunctivitis followed by pneumonia and arthritis was observed in beef calves of a managed herd. No Moraxella spp. or bacteria other than Mycoplasma spp. were obtained from conjunctival and nasal swabs. A strategy was designed for characterization of bovine mycoplasmas at species and strain level on the basis of a combination of molecular tools and the immunoblotting method. The strategy made it possible to rapidly assign the bacterium responsible for this outbreak to one of the phylogenetic clusters of bovine mycoplasmas delineated in this study and then to identify it as Mycoplasma bovis. The strain, designated Sar 1, showed a 100% 16S rDNA sequence identity with two European strains (120/81 and MC3386) isolated in Germany and Ireland, respectively, and hosts a vsp gene analog to the vspA, vsp422-4, and vsp422-8 genes of the M. bovis reference strain PG45T and of the field strain 422. The use of a cross-reactive rabbit serum developed against the Mycoplasma agalactiae immunodominant antigen P48 confirmed the molecular findings. The immunological response of calves against M. bovis was also investigated. This is the first report on the occurrence of M. bovis on the Island of Sardinia (Italy).

Animals↗

PubMed1995

Immunohistochemical and pathological study of Mycoplasma bovis-associated lung abscesses in calves.

Out of 45 cases of fatal chronic pneumonia in calves examined for Mycoplasma bovis infection from February to July 1994, 11 cases with pulmonary abscesses that were culture positive for M. bovis were encountered. The cases were studied in detail using a recently developed monoclonal antibody-based immunoperoxidase technique. Mycoplasma bovis organisms were detected in specific locations at all stages of abscessation observed. In bronchioles or terminal airways within which abscesses developed, M. bovis was located at the epithelial surface and in close association with infiltrating neutrophils and macrophages. Abscessed airways that had lost the epithelium were encapsulated and were seen as coagulative necrotic foci that stained intensely for M. bovis, especially at the periphery. Some foci stained weakly and such might have been resolving lesions. Mycoplasma bovis was also demonstrated at sites of mild mononuclear cell infiltration in the livers and kidneys of 2 calves. The mycoplasma was detected within bile ducts in the liver and in the tubular epithelium of the kidney. Abscesses not staining for M. bovis, presumably caused by other pathogens, were seen concurrently with M. bovis-associated abscesses in some lungs. Thirteen other M. bovis-positive cases showed no abscesses, possibly indicating heterogeneity among M. bovis strains. Three other cases with abscesses were negative for M. bovis by culture and immunoperoxidase staining. The monoclonal antibody-based immunohistochemical technique is efficient for specific detection of M. bovis in cases of enzootic pneumonia of calves with or without abscessation. Mycoplasma bovis is implicated in the pathogenesis of lung abscesses in some calves.

Animals↗

PubMed1998

Genital mycoplasmas stimulate tumor necrosis factor-alpha and inducible nitric oxide synthase production from a murine macrophage cell line.

Ureaplasma urealyticum and Mycoplasma hominis, two genital mycoplasmas, are the most common organisms isolated in the perinatal period and both either cause or are associated with poor perinatal outcomes. We speculate that these microbes could increase inflammation by stimulating macrophages to produce tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase because of their propensity to interact with the host's immune system. To test this hypothesis, RAW 264.7 cells, a murine macrophage cell line, were coincubated for 16 h with either U. urealyticum or M. hominis, and LPS and sterile broth were used as controls. Lipopolysaccharide (LPS) and both mycoplasmas induced TNF-alpha production, which was concentration-dependent, whereas sterile broth had little effect. TNF-alpha production was not inhibited by the addition of polymyxin B, excluding the possibility of contaminating endotoxin in this effect. Inducible nitric oxide synthase was produced only in the presence of recombinant inteferon-gamma. We conclude that both viable and nonviable U. urealyticum and M. hominis are capable of TNF-alpha induction from murine macrophages and that LPS is not involved in this event. Also, the genital mycoplasmas are capable of stimulating inducible nitric oxide synthase production from murine macrophages. We speculate that the genital mycoplasmas produce perinatal disease by producing proinflammatory mediators by their interaction with inflammatory cells and either induce or act as a catalyst and augment inflammation which in turn leads to a poor pregnancy outcome.

Animals↗

PubMed2000

Late abnormal findings on high-resolution computed tomography after Mycoplasma pneumonia.

BACKGROUND: The clinical course of Mycoplasma pneumonia is typically mild and self-limited. There are, however, several case reports of severe complication following this illness with considerable morbidity and mortality. OBJECTIVES: This study was conducted to investigate, using high-resolution computed tomography (HRCT), the long-term pulmonary structural abnormalities after Mycoplasma pneumonia and to identify risk factors (chest radiograph findings, antibody titers, and host factors) that might increase the likelihood of developing the sequelae. METHODS: Thirty-eight children requiring hospitalization attributable to Mycoplasma pneumonia were recruited by the retrospective examination of hospital records. They underwent HRCT after an interval of 1.0 to 2. 2 years. A control group of 17 children with the history of Mycoplasma upper respiratory infection was also studied after a similar interval. RESULTS: Abnormal HRCT findings were present in 37% (14/38) of the pneumonia group, compared with 12% (2/17) of the control group. The abnormalities in the pneumonia group, which appeared alone or in combination, included mosaic perfusion (n = 12), bronchiectasis (n = 8), bronchial wall thickening (n = 4), decreased vascularity (n = 1), and air trapping on expiratory scan (9 of 29 tested). The area affected by these abnormalities, usually involving 2 or more lobes, corresponded in all cases to the location of the infiltrate on chest radiograph at the time of pneumonia. Between subjects with abnormal HRCT (n = 14) and normal HRCT (n = 24) in the pneumonia group, significant differences were observed in age at the time of pneumonia (mean +/- standard deviation: 5.3 +/- 2. 0 years vs 7.7 +/- 3.4 years) and peak antimycoplasma antibody titer (geometric mean [range of 1 standard deviation]; 1:7943 [3126-19 953] vs 1:3093 [832-11 482]). CONCLUSIONS: We conclude that a considerable proportion of children with history of Mycoplasma pneumonia have abnormal findings on HRCT, suggestive of small airway obstruction and that younger age and higher antibody titer at the time of pneumonia may be risk factors for these sequelae.

Antibodies, Bacterial↗

PubMed2004

Inactivation, storage, and PCR detection of Mycoplasma on FTA filter paper.

We evaluated the feasibility of using Flinders Technology Associates (FTA) filter paper for the inactivation and storage of mycoplasma DNA templates and their detection by the polymerase chain reaction (PCR). FTA paper is a cotton-based cellulose membrane containing lyophilized chemicals that lyses most types of bacteria and viruses. Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) cultures were spotted at various volumes on the filter paper and stored at different temperatures for various periods of time before performing PCR. MG and MS were readily detected at all time frames (1-60 days) independent of the volume applied (1-100 microl) or storage temperature (4 C-41 C). Sensitivity and specificity of the FTA-PCR were comparable to the standard diagnostic PCR, allowing the detection of MG/MS in field samples without interference by nontargeted mycoplasma. Analysis of 193 field samples by both methods showed nearly 100% agreement with serology and culture results. The long-term DNA stability at a wide range of temperatures makes the FTA cards a good alternative for collecting and simultaneously inactivating mycoplasma. It also offers the convenience of storage and transport of DNA in a cost-effective manner for further molecular analysis, such as restriction enzyme length polymorphism and nucleotide sequencing.

Bacteriological Techniques↗

PubMed1996

Mycoplasma bovis as an agent of mastitis, pneumonia, arthritis and genital disorders in cattle.

Bovine diseases due to Mycoplasma bovis can cause considerable economic losses in cattle production. While the pathogen is principally responsible for therapy-resistant mastitis on large dairy farms, on smaller farms the typical mycoplasma diseases are calf pneumonia and arthritis. Moreover, the pathogen is able to cause genital disorders. M. bovis infection can be controlled effectively only if appropriate measures are implemented at the earliest possible stage. Since immunoprophylaxis and antibiotic treatment are known to be ineffective, control measures must include the introduction of strict hygiene standards, the restriction of animal movement out of infected herds and the culling of clinically diseased animals and shedders of the mycoplasma (the latter only in the case of mastitis and genital disorders). In this review, symptoms of the various diseases caused by M. bovis are described and characteristics of the course of infection are outlined. To clarify the origin and spread of the infection, the authors describe the main properties and reservoirs of the pathogen and summarise experimental evidence on modes of transmission to susceptible organs. As effective diagnosis is a prerequisite for the introduction of early control measures, the advantages and disadvantages of currently used diagnostic methods are discussed in detail. It is a serious shortcoming if testing for mycoplasmas is not included in routine bacterial examination of clinical samples. As a consequence, some M. bovis infections will remain undetected and outbreaks cannot be controlled properly. Finally, practical recommendations are given for prevention and control, including the formation of mycoplasma-free herds.

Animals↗

PubMedDate not supplied

Adenosine deaminase activity and free IL-2 receptor levels in serum from patients with mycoplasma pneumonia.

Adenosine deaminase (ADA) activity and free interleukin (IL)-2 receptor levels were assayed in serum samples from patients with mycoplasma and bacterial pneumonia to evaluate the usefulness of these parameters in distinguishing between these diseases at an early stage. Serum ADA and free IL-2 receptor levels in patients with mycoplasma pneumonia (32.4 +/- 9.2 U/l, 960 +/- 204 U/ml) were significantly higher than those in patients with bacterial pneumonia (12.5 +/- 3.3 U/l, 425 +/- 86 U/ml) and in healthy controls (14.0 +/- 3.4 U/l, 286 +/- 49 U/ml) (p less than 0.001). Of the 20 mycoplasma pneumonia cases, 19 showed increased levels of ADA over 20.8 U/l; in 17 of the 19, the increase of ADA was seen before the elevation of the specific antibody to Mycoplasma pneumoniae. In contrast, serum ADA levels in all 20 cases of bacterial pneumonia were lower than 20.8 U/l. There results indicate that assays for serum ADA and free IL-2 receptor levels are useful in distinguishing between bacterial and mycoplasma pneumonia at an early stage.

Adenosine Deaminase↗

PubMed2003

T lymphocyte responses are critical determinants in the pathogenesis and resistance to mycoplasma respiratory disease.

Mycoplasmas are responsible for many human and animal respiratory diseases and have a tremendous economic and health impact worldwide. Currently, there is no vaccine against any animal or human mycoplasma species. Despite the vast amount of research, the mechanisms that control hosts' resistance and susceptibility to mycoplasma infection remains unclear. Immune responses, particularly T lymphocyte responses, are believed to be critical players in the pathogenesis of mycoplasma disease. In this review, we will highlight the potential roles of T lymphocytes as influential mediators in animal and human mycoplasma pathogenesis and resistance infection.

Animals↗