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1.
Am J Obstet Gynecol ; 185(1): 163-5, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11483922

ABSTRACT

OBJECTIVE: We sought to determine the midtrimester prevalence of Mycoplasma genitalium in women who had subsequent spontaneous preterm birth. STUDY DESIGN: In a prospective study of lower genital tract infections, we identified 127 women who subsequently had spontaneous preterm birth. Vaginal samples were obtained between 21 and 25 weeks' gestation for pH, for bacterial vaginosis Gram stain, and cultures that yielded Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis. M genitalium was identified by using validated polymerase chain reaction (PCR) primers, and the results were compared to pregnancy outcomes. RESULTS: Of 124 women with spontaneous preterm births, only five (3.9%) had PCR assays positive forM genitalium. The mean +/- SD delivery gestational age was similar for women with a positive PCR (34.6 +/- 2.2 weeks) and a negative PCR (34.0 +/- 2.7 weeks) (P =.62). None of the women with positive PCR results tested positive for any other sexually transmitted disease, whereas 36 (30%) women with negative PCR results tested positive. CONCLUSIONS: The occurrence of M genitalium in the vagina at midtrimester is infrequent in women with subsequent spontaneous preterm birth.


Subject(s)
Gestational Age , Mycoplasma Infections , Obstetric Labor, Premature/microbiology , Vaginosis, Bacterial/microbiology , Animals , Chlamydia trachomatis/isolation & purification , DNA, Bacterial/analysis , Female , Humans , Mycoplasma/genetics , Mycoplasma/isolation & purification , Mycoplasma Infections/microbiology , Neisseria gonorrhoeae/isolation & purification , Polymerase Chain Reaction , Pregnancy , Pregnancy Trimester, Second , Prospective Studies , Trichomonas vaginalis/isolation & purification , Vagina/microbiology
2.
J Allergy Clin Immunol ; 107(4): 595-601, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11295645

ABSTRACT

BACKGROUND: Asthma is a prevalent disease with marked effects on quality of life and economic societal burden. However, the cause of asthma and its pathophysiology are not completely defined. Recently, the possibility that chronic infection may play a role has been suggested. OBJECTIVE: We sought to define the association between Mycoplasma and Chlamydia species and chronic asthma. METHODS: We performed a comparison study of asthmatic patients and normal control subjects. Fifty-five patients with chronic stable asthma were compared with 11 normal control subjects by using PCR, culture, and serology for Mycoplasma species, Chlamydia species, and viruses from the nasopharynx, lung, and blood. Bronchoalveolar lavage cell count and differential, as well as tissue morphometry, were also evaluated. Computer-generated scoring for the degree of chronic sinusitis in asthmatic patients was additionally evaluated. RESULTS: Thirty-one of 55 asthmatic patients had positive PCR results for Mycoplasma (n = 25) or Chlamydia species (n = 6), which were mainly found on lung biopsy specimens or in lavage fluid. Only 1 of 11 normal control subjects had positive PCR results for Mycoplasma species. The distinguishing phenotype between asthmatic patients with positive and negative PCR results was the significantly greater number of tissue mast cells in the group with positive results. CONCLUSION: A significant number of patients with chronic stable asthma demonstrate the presence of Mycoplasma species, Chlamydia species, or both in their airways, with the distinguishing feature of increased mast cell number. These findings need further delineation but may help us to understand the pathophysiology of asthma and new treatment options.


Subject(s)
Asthma/etiology , Chlamydia Infections/complications , Mycoplasma Infections/complications , Adult , Antibodies, Bacterial/blood , Asthma/microbiology , Asthma/virology , Chlamydia/immunology , Chlamydia/isolation & purification , Chronic Disease , Female , Humans , Male , Mycoplasma/immunology , Mycoplasma/isolation & purification , Polymerase Chain Reaction
3.
JAMA ; 285(5): 601-5, 2001 Feb 07.
Article in English | MEDLINE | ID: mdl-11176866

ABSTRACT

During the past 2 decades, new infectious diseases have appeared and old ones previously thought to be controlled have reemerged. New and reemerging infectious agents will continue to pose serious threats well into the 21st century. The prediction that the threat of infectious disease may not diminish is supported by evidence that infectious agents cause many chronic diseases and cancer of previous unknown etiology. Moreover, the utility of existing antimicrobial agents is rapidly eroding, tipping the balance in favor of multidrug-resistant pathogens, and there appear to be few, if any, new classes of drugs currently in clinical development. The need for research directed toward development of new antibiotics has never been greater. Advances in research technologies and microbial genome sequencing in the past decade have led to identification of a large number of new targets. Functional genomics and integrative biology should validate these targets and provide the best opportunity for developing effective new therapies, improved diagnostic techniques, and better tools to understand host-pathogen interactions.


Subject(s)
Anti-Bacterial Agents , Communicable Diseases/drug therapy , Drug Resistance, Microbial , Research/trends , Animals , Chemistry, Pharmaceutical , Computational Biology , Forecasting , Genomics , Humans , Xenobiotics
4.
Nature ; 407(6805): 757-62, 2000 Oct 12.
Article in English | MEDLINE | ID: mdl-11048724

ABSTRACT

The comparison of the genomes of two very closely related human mucosal pathogens, Mycoplasma genitalium and Mycoplasma pneumoniae, has helped define the essential functions of a self-replicating minimal cell, as well as what constitutes a mycoplasma. Here we report the complete sequence of a more distant phylogenetic relative of those bacteria, Ureaplasma urealyticum (parvum biovar), which is also a mucosal pathogen of humans. It is the third mycoplasma to be sequenced, and has the smallest sequenced prokaryotic genome except for M. genitalium. Although the U. urealyticum genome is similar to the two sequenced mycoplasma genomes, features make this organism unique among mycoplasmas and all bacteria. Almost all ATP synthesis is the result of urea hydrolysis, which generates an energy-producing electrochemical gradient. Some highly conserved eubacterial enzymes appear not to be encoded by U. urealyticum, including the cell-division protein FtsZ, chaperonins GroES and GroEL, and ribonucleoside-diphosphate reductase. U. urealyticum has six closely related iron transporters, which apparently arose through gene duplication, suggesting that it has a kind of respiration system not present in other small genome bacteria The genome is only 25.5% G+C in nucleotide content, and the G+C content of individual genes may predict how essential those genes are to ureaplasma survival.


Subject(s)
Genome, Bacterial , Ureaplasma urealyticum/genetics , Adenosine Triphosphate/metabolism , Biological Evolution , DNA, Bacterial , Humans , Iron/metabolism , Molecular Sequence Data , Sequence Analysis, DNA , Ureaplasma urealyticum/classification , Ureaplasma urealyticum/metabolism , Ureaplasma urealyticum/pathogenicity
5.
Lab Anim Sci ; 49(4): 385-94, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10480643

ABSTRACT

BACKGROUND AND PURPOSE: Sendai virus infection in rats is an excellent model for studying development and role of host defenses throughout the respiratory tract after this infection. Therefore, development of serum antibody responses and disease were studied. METHODS: Forty-two anesthetized pathogen-free 3- to 4- week-old LEW/NCr rats were inoculated intranasally with Sendai virus. At postinoculation days 0, 2, 3, 5, 8, 10, and 14, rats were euthanized by administration of a pentobarbital sodium overdose followed by exsanguination. Serum was obtained from all animals, and nasal wash and bronchoalveolar lavage specimens were collected during selected experiments. An ELISPOT assay was used to measure numbers of Sendai virus-specific antibody-forming cells in respiratory tract lymphoid tissue. RESULTS: Recovery from disease and clearance of virus from respiratory tract tissues coincided with development of serum antibody responses. Upper respiratory tract lymph nodes were the initial and major sites of appearance of antibody-forming cells. Immunoglobulin G was the predominant subtype of these cells during recovery from the infection and in rats resistant to infection. Passive transfer of antisera or specific IgG protected the lower but not the upper respiratory tract. CONCLUSIONS: Circulating components of immunity have a major role in resistance and recovery from disease in the lower respiratory tract, whereas local responses are likely involved in protection of the upper respiratory tract. Local lymphoid tissues are the major production sites of IgG, which contributes to resistance to and recovery from respiratory tract diseases.


Subject(s)
Antibodies, Viral/analysis , Respiratory Tract Diseases/virology , Respirovirus Infections/virology , Respirovirus/immunology , Animals , Antibodies, Viral/blood , Bronchoalveolar Lavage Fluid/immunology , Immunity, Innate , Immunization, Passive , Immunoglobulin A/analysis , Immunoglobulin G/analysis , Lung/immunology , Lymphoid Tissue/immunology , Male , Nose/immunology , Rats , Rats, Inbred Lew , Respiratory System/immunology , Respiratory System/virology , Respiratory Tract Diseases/immunology , Respirovirus Infections/immunology , Therapeutic Irrigation
6.
Emerg Infect Dis ; 5(1): 164-7, 1999.
Article in English | MEDLINE | ID: mdl-10081687

ABSTRACT

Mycoplasma penetrans, a rare bacterium so far only found in HIV-infected persons, was isolated in the blood and throat of a non-HIV-infected patient with primary antiphospholipid syndrome (whose etiology and pathogenesis are unknown).


Subject(s)
Antiphospholipid Syndrome/complications , Bacteremia/complications , Mycoplasma Infections/complications , Mycoplasma penetrans/isolation & purification , Adolescent , Female , Humans , Mycoplasma Infections/diagnosis
7.
Pediatr Infect Dis J ; 17(10): 865-71, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9802626

ABSTRACT

OBJECTIVE: To compare the safety and efficacy of azithromycin with amoxicillin/clavulanate or erythromycin for the treatment of community-acquired pneumonia, including atypical pneumonia caused by Mycoplasma pneumoniae and Chlamydia pneumoniae. METHODS: Multicenter, parallel group, double blind trial in which patients 6 months to 16 years of age with community-acquired pneumonia were randomized 2:1 to receive either azithromycin for 5 days or conventional therapy for 10 days (amoxicillin/clavulanate if < or =5 years of age or erythromycin estolate if >5 years of age). Patients from 23 geographically diverse sites were evaluated for clinical outcomes and/or adverse events at Days 3 to 5, Days 15 to 19 and 4 to 6 weeks posttherapy. Microbiology (culture or polymerase chain reaction) was done at baseline and Days 15 to 19 for bacteria, Chlamydia pneumoniae and Mycoplasma pneumoniae. Serology for C. pneumoniae and M. pneumoniae was done at baseline and 4 to 6 weeks posttherapy. RESULTS: Of 456 patients enrolled during 17 consecutive months, 420 were evaluable. Clinical success at Study Days 15 to 19 was 94.6% in the azithromycin group and 96.2% in the comparative treatment group (P = 0.735) and at 4 to 6 weeks posttherapy 90.6 and 87.1%, respectively (P = 0.330). Evidence of infection was identified in 46% of 420 evaluable patients (1.9% bacteria, 29.5% M. pneumoniae and 15% C. pneumoniae). Microbiologic eradication was 81% for C. pneumoniae and 100% for M. pneumoniae in the azithromycin group vs. 100 and 57%, respectively, in the comparator group. Treatment-related adverse events occurred in 11.3% of the azithromycin group and 31% in the comparator group (P < 0.05). CONCLUSION: Azithromycin used once daily for 5 days produced a satisfactory therapeutic outcome similar to those of amoxicillin/clavulanate or erythromycin given three times a day for 10 days for treatment of community-acquired pneumonia. Azithromycin had significantly fewer side effects than comparator drugs.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Azithromycin/therapeutic use , Pneumonia, Bacterial/drug therapy , Adolescent , Amoxicillin-Potassium Clavulanate Combination/therapeutic use , Child , Child, Preschool , Chlamydia Infections/drug therapy , Chlamydophila pneumoniae , Communicable Diseases , Double-Blind Method , Drug Therapy, Combination/therapeutic use , Erythromycin/therapeutic use , Female , Humans , Infant , Male , Mycoplasma pneumoniae , Pneumonia, Mycoplasma/drug therapy
8.
Am J Respir Crit Care Med ; 158(3): 998-1001, 1998 Sep.
Article in English | MEDLINE | ID: mdl-9731038

ABSTRACT

Infection with Mycoplasma pneumoniae has been shown to exacerbate asthma in humans. However, the role of M. pneumoniae in the pathogenesis of chronic asthma has not been defined. Eighteen asthmatics with chronic, stable asthma and 11 nonasthmatic control subjects underwent evaluation of the upper and lower airways and serologic analysis to determine the presence of M. pneumoniae, Chlamydia pneumoniae, and seven respiratory viruses through culture, enzyme-linked immunoassay (EIA) and polymerase chain reaction (PCR). M. pneumoniae was detected by PCR in 10 of 18 asthmatics and one of 11 control subjects (p = 0.02). In nine of the 10 patients, the organism was detected in bronchoalveolar lavage or bronchial biopsies. Seven of 18 asthmatics and one of 11 control subjects were also positive for M. fermentans and M. genitalium by PCR. All patients' cultures, EIAs, and serology were negative for M. pneumoniae. All PCR and cultures were negative for C. pneumoniae, and all EIAs for respiratory viruses were negative in all subjects. Nine asthmatics and one control subject exhibited positive serology for C. pneumoniae (p = 0.05). M. pneumoniae was present in the lower airways of chronic, stable asthmatics with greater frequency than control subjects, and may play a role in the pathogenesis of chronic asthma.


Subject(s)
Asthma/microbiology , Bronchi/microbiology , Mycoplasma pneumoniae/isolation & purification , Adult , Biopsy , Bronchi/pathology , Bronchoalveolar Lavage Fluid/microbiology , Chlamydia Infections/diagnosis , Chlamydophila pneumoniae/isolation & purification , Chronic Disease , Female , Humans , Influenza A virus/isolation & purification , Influenza B virus/isolation & purification , Influenza, Human/diagnosis , Male , Mycoplasma/classification , Mycoplasma/isolation & purification , Mycoplasma fermentans/isolation & purification , Parainfluenza Virus 1, Human/isolation & purification , Parainfluenza Virus 2, Human/isolation & purification , Parainfluenza Virus 3, Human/isolation & purification , Pneumonia, Mycoplasma/complications , Pneumonia, Mycoplasma/diagnosis , Respiratory Syncytial Virus Infections/diagnosis , Respiratory Syncytial Viruses/isolation & purification , Respirovirus Infections/diagnosis , Rubulavirus Infections/diagnosis
9.
Emerg Infect Dis ; 4(3): 475-87, 1998.
Article in English | MEDLINE | ID: mdl-9716980

ABSTRACT

Powerful diagnostic technology, plus the realization that organisms of otherwise unimpressive virulence can produce slowly progressive chronic disease with a wide spectrum of clinical manifestations and disease outcomes, has resulted in the discovery of new infectious agents and new concepts of infectious diseases. The demonstration that final outcome of infection is as much determined by the genetic background of the patient as by the genetic makeup of the infecting agent is indicating that a number of chronic diseases of unknown etiology are caused by one or more infectious agents. One well-known example is the discovery that stomach ulcers are due to Helicobacter pylori. Mycoplasmas may cause chronic lung disease in newborns and chronic asthma in adults, and Chlamydia pneumoniae, a recently identified common cause of acute respiratory infection, has been associated with atherosclerosis. A number of infectious agents that cause or contribute to neoplastic diseases in humans have been documented in the past 6 years. The association and causal role of infectious agents in chronic inflammatory diseases and cancer have major implications for public health, treatment, and prevention.


Subject(s)
Arteriosclerosis/microbiology , Inflammation/microbiology , Lung Diseases/microbiology , Neoplasms/microbiology , Animals , Chronic Disease , Disease Models, Animal , Humans
10.
Infect Immun ; 66(8): 3485-91, 1998 Aug.
Article in English | MEDLINE | ID: mdl-9673224

ABSTRACT

Current evidence suggests that host defense in respiratory mycoplasmosis is dependent on both innate and humoral immunity. To further delineate the roles of innate and adaptive immunity in antimycoplasmal defenses, we intranasally infected C3H/HeSnJ-scid/scid (C3H-SCID), C3H/HeSnJ (C3H), C57BL/6J-scid/scid (C57-SCID), and C57BL/6N (C57BL) mice with Mycoplasma pulmonis and at 14 and 21 days postinfection performed quantitative cultures of lungs and spleens, quantification of lung lesions, and histopathologic assessments of all other major organs. We found that numbers of mycoplasmas in lungs were associated with genetic background (C3H susceptible, C57BL resistant) rather than functional state of adaptive immunity, indicating that innate immunity is the main contributor to antimycoplasmal defense of the lungs. Extrapulmonary dissemination of mycoplasmas with colonization of spleens and histologic lesions in multiple organs was a common occurrence in all mice. The absence of adaptive immune responses in severe combined immunodeficient (SCID) mice resulted in increased mycoplasmal colonization of spleens and lesions in extrapulmonary sites, particularly spleens, hearts, and joints, and also reduced lung lesion severity. The transfer of anti-M. pulmonis serum to infected C3H-SCID mice prevented extrapulmonary infection and disease, while the severity of lung lesions was restored by transfer of naive spleen cells to infected C3H-SCID mice. Collectively, our results strongly support the conclusions that innate immunity provides antimycoplasmal defense of the lungs and humoral immunity has the major role in defense against systemic dissemination of mycoplasmal infection, but cellular immune responses may be important in exacerbation of mycoplasmal lung disease.


Subject(s)
Lung Diseases/immunology , Mycoplasma Infections/immunology , Animals , Antibodies, Bacterial/immunology , Bacterial Vaccines/immunology , Disease Models, Animal , Female , Immunity, Active/immunology , Immunity, Innate/genetics , Immunity, Innate/immunology , Immunization, Passive , Lung/immunology , Lung/microbiology , Lung/pathology , Lung Diseases/genetics , Lung Diseases/microbiology , Lung Diseases/pathology , Male , Mice , Mice, Inbred C3H , Mice, Inbred C57BL , Mice, SCID , Mycoplasma Infections/genetics , Mycoplasma Infections/microbiology , Mycoplasma Infections/pathology , Severe Combined Immunodeficiency/immunology , Spleen/cytology , Spleen/immunology
11.
Clin Infect Dis ; 26(1): 103-10, 1998 Jan.
Article in English | MEDLINE | ID: mdl-9455517

ABSTRACT

Since the 1950s the U.S. military has used intramuscular injections of benzathine penicillin G (BPG) to control outbreaks of respiratory disease. In an effort to find an alternative prophylaxis, a randomized field trial was conducted among 1,016 male U.S. Marine trainee volunteers at high risk for respiratory disease. Participants were evaluated for evidence of acute respiratory infection by serological tests on pretraining and posttraining sera (63 days apart). Oral azithromycin prophylaxis (500 mg/w) outperformed BPG, preventing infection from Streptococcus pyogenes (Efficacy [E] = 84%; 95% confidence interval [CI], 63%-93%), Streptococcus pneumoniae (E = 80%; 95% CI, 50%-92%), Mycoplasma pneumoniae (E = 64%; 95% CI, 25%-83%), and Chlamydia pneumoniae (E = 58%; 95% CI, 15%-79%) in comparison with results in a no-treatment group. Azithromycin group subjects reported few side effects and less respiratory symptoms than the BPG and no-treatment groups. According to serological tests, oral azithromycin is an effective alternative prophylaxis to BPG for military populations.


Subject(s)
Azithromycin/therapeutic use , Respiratory Tract Infections/prevention & control , Administration, Oral , Adult , Azithromycin/administration & dosage , Azithromycin/adverse effects , Humans , Male , Penicillin G Benzathine/therapeutic use , Pharynx/microbiology , Streptococcus/isolation & purification
12.
Infect Immun ; 66(1): 272-9, 1998 Jan.
Article in English | MEDLINE | ID: mdl-9423868

ABSTRACT

Murine chronic respiratory disease is characterized by persistent colonization of tracheal and bronchial epithelial cell surfaces by Mycoplasma pulmonis, submucosal and intraluminal immune and inflammatory cells, and altered airway activity. To determine the direct effect of M. pulmonis upon transepithelial ion transport in the absence of immune and inflammatory cell responses, primary mouse tracheal epithelial cell monolayers (MTEs) were apically infected and assayed in Ussing chambers. M. pulmonis-infected MTEs, but not those infected with a nonmurine mycoplasma, demonstrated reductions in amiloride-sensitive Na+ absorption, cyclic AMP, and cholinergic-stimulated Cl- secretion and transepithelial resistance. These effects were shown to require interaction of viable organisms with the apical surface of the monolayer and to be dependent upon organism number and duration of infection. Altered transport due to M. pulmonis was not merely a result of epithelial cell death as evidenced by the following: (i) active transport of Na+ and Cl-, albeit at reduced rates; (ii) normal cell morphology, including intact tight junctions, as demonstrated by electron microscopy; (iii) maintenance of a mean transepithelial resistance of 440 omega/cm2; and (iv) lack of leakage of fluid from the basolateral to the apical surface of the monolayer. Alteration in epithelial ion transport in vitro is consistent with impaired pulmonary clearance and altered airway function in M. pulmonis-infected animals. Furthermore, the ability of M. pulmonis to alter transport without killing the host cell may explain its successful parasitism and long-term persistence in the host. Further study of the MTE-M. pulmonis model should elucidate the molecular mechanisms which mediate this reduction in transepithelial ion transport.


Subject(s)
Chlorides/metabolism , Mycoplasma Infections/metabolism , Sodium/pharmacokinetics , Amiloride/metabolism , Animals , Cell Polarity , Cell Size , Cells, Cultured , Cholinergic Agents/metabolism , Cyclic AMP/metabolism , Epithelial Cells/cytology , Epithelial Cells/ultrastructure , Ion Transport , Mice , Species Specificity , Tight Junctions/metabolism , Tight Junctions/ultrastructure , Trachea/cytology
13.
Mil Med ; 162(8): 524-6, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9271902

ABSTRACT

From August 1993 through April 1994, U.S. Marines (98% male, median age 20 years) who were hospitalized with radiographically confirmed pneumonia were prospectively studied for evidence of acute Mycoplasma pneumoniae infection. Overall, 32 (36.4%) of the 88 patients with paired sera had evidence of acute infection by an elevated immunoglobulin M titer (22.7%), a 4-fold rise in immunoglobulin G titer (9.1%), a positive polymerase chain reaction result (11.1%), and/or a positive culture (5.8%). No specific symptoms or clinical findings were strong predictors of M. pneumoniae infection. Among patients with evidence of acute M. pneumoniae infection, admitting clinicians chose other pathogens as more likely etiologic agents 46.4% of the time, and over the course of the hospitalization, 10% of patients failed to receive appropriate antibiotics. These data indicate that M. pneumoniae may cause a high proportion of pneumonias among military personnel and should be considered in empiric treatment and prophylaxis.


Subject(s)
Hospitalization/statistics & numerical data , Military Personnel , Pneumonia, Mycoplasma/virology , Acute Disease , Adolescent , Adult , California , Female , Humans , Incidence , Male , Middle Aged , Naval Medicine , Pneumonia, Mycoplasma/diagnostic imaging , Radiography , Risk Factors , Seasons
16.
J Clin Microbiol ; 35(4): 992-4, 1997 Apr.
Article in English | MEDLINE | ID: mdl-9157171

ABSTRACT

Mycoplasma hominis is a commensal in the genital tract of women and has been associated with urogenital and extragenital infections. However, central nervous system infections with this organism in adults are very rare. Here we describe the recovery of M. hominis from a brain abscess associated with a postpartum infection. Seroconversion to the isolated strain was detected by both a metabolic inhibition test and an immunoblotting assay. This case demonstrates the pathogenic potential of M. hominis and the need for rapid recognition of the organism so that appropriate chemotherapeutic intervention can occur.


Subject(s)
Abscess/microbiology , Brain Diseases/microbiology , Mycoplasma Infections/microbiology , Mycoplasma hominis/isolation & purification , Postpartum Period , Adult , Female , Humans , Mycoplasma Infections/physiopathology
17.
Diagn Microbiol Infect Dis ; 27(4): 117-22, 1997 Apr.
Article in English | MEDLINE | ID: mdl-9154407

ABSTRACT

The Etest was compared with microbroth dilution for performing in vitro susceptibility tests in 38 isolates of Mycoplasma hominis chosen to represent a wide range of MIC values. MIC50s were 4 micrograms/ml for both methods; whereas, MIC90s were 64 and > 256 micrograms/ml for broth and Etest, respectively. Etest MICs determined on SP 4 agar were usually two or more dilutions greater than microbroth MICs in SP 4 broth, and values were prone to change by one to two dilutions when different inoculum densities were used. Inocula of 10(5) to 10(6) color-changing units/ml gave the most consistently readable MICs, with discrete colony formation surrounding the ellipse. Etest MICs for 5 isolates tested a second time at the same inoculum on SP 4 agar agreed with the original value within 1 dilution. For 12 isolates tested on A 8 agar simultaneously with SP 4 agar, MICs for 10/12 agreed within one dilution; whereas, in the other 2 isolates, MICs varied by two dilutions. These findings suggest that the Etest, when properly controlled, can be used to determine tetracycline MICs for M. hominis.


Subject(s)
Mycoplasma hominis/drug effects , Tetracycline Resistance , Microbial Sensitivity Tests , Mycoplasma Infections/drug therapy , Mycoplasma hominis/isolation & purification , Mycoplasma hominis/metabolism
18.
Infect Immun ; 64(12): 5326-31, 1996 Dec.
Article in English | MEDLINE | ID: mdl-8945584

ABSTRACT

Mouse strains differ markedly in resistance to Mycoplasma pulmonis infection, and investigation of these differences holds much promise for understanding the mechanisms of antimycoplasmal host defenses. To determine the potential genetic diversity of resistance to disease in murine respiratory mycoplasmosis (MRM) and to select disease-resistant and nonresistant mouse strains for further genetic analysis, we screened 17 inbred mouse strains of various Bcg and H-2 genotypes for resistance to M. pulmonis. Mice were inoculated intranasally with 10(4) CFU of M. pulmonis UAB CT and evaluated at 21 days postinfection for severities of the four histologic lung lesions characteristic of MRM: alveolar exudate, airway exudate, airway epithelial hyperplasia, and lymphoid infiltrate. On the basis of these assessments of MRM severity, one group of mouse strains was found to be extremely resistant to disease (C57BR/cdJ, C57BL/6NCr, C57BL/10ScNCr, and C57BL/6J). The remaining strains of mice (C57L/J, SJL/NCr, BALB/cAnNCr, A/JCr, C3H/HeJ, SWR/J, AKR/NCr, CBA/NCr, C58/J, DBA/2NCr, C3H/HeNCr, C3HeB/FeJ, and C3H/HeJCr) developed disease of widely varying severities. Furthermore, strains in the group with more disease varied in pattern of lesion severity. While the severities of all four lesions were correlated in most mouse strains, this was not always true. DBA/2NCr mice had one of the highest scores for alveolar exudate, only a moderate score for airway exudate, and significantly lower scores for both airway epithelial hyperplasia and lymphoid infiltrate than all other strains susceptible to lung disease. DBA/2NCr mice had one of the highest mortality rates. We concluded that resistance to MRM is a complex trait. The observed differences in lung disease severity could not be explained by known differences at the Bcg or H-2 locus in the strains of mice we studied.


Subject(s)
Mycoplasma Infections/genetics , Mycoplasma , Pneumonia, Bacterial/genetics , Animals , Mice , Mice, Inbred Strains , Species Specificity
19.
Clin Diagn Lab Immunol ; 3(6): 774-8, 1996 Nov.
Article in English | MEDLINE | ID: mdl-8914774

ABSTRACT

One of the major surface structures of Ureaplasma urealyticum recognized by antibodies of patients during infection is the MB antigen. Previously, we showed by Western blot (immunoblot) analysis that any one of the anti-MB monoclonal antibodies (MAbs) 3B1.5, 5B1.1, and 10C6.6 could block the binding of patient antibodies to MB. Subsequent DNA sequencing revealed that a unique six-amino-acid direct tandem repeat region composed the carboxy two-thirds of this antigen. In the present study, using antibody-reactive peptide scanning of this repeat region, we demonstrated that the amino acids defining the epitopes for MAbs 3B1.5 5B1.1 and 10C6.6 are EQP, GK, and KEQPA, respectively. Peptide scanning analysis of an infected patient's serum antibody response showed that the dominant epitope was defined by the sequence PAGK. Mapping of these continuous epitopes revealed overlap between all MAb and patient polyclonal antibody binding sites, thus explaining the ability of a single MAb to apparently block all polyclonal antibody binding sites. We also show that a single amino acid difference in the sequence of the repeats of serovars 3 and 14 accounts for the lack of reactivity with serovar 14 of two of the serovar 3-specific MAbs. Finally, the data demonstrate the need to obtain the sequences of the mba genes of all serovars before an effective serovar-specific antibody detection method can be developed.


Subject(s)
Bacterial Proteins/chemistry , Epitope Mapping , Epitopes/chemistry , Repetitive Sequences, Nucleic Acid/immunology , Ureaplasma urealyticum/immunology , Amino Acid Sequence , Antigens, Bacterial/chemistry , Humans , Molecular Sequence Data
20.
Int J Syst Bacteriol ; 46(3): 827-9, 1996 Jul.
Article in English | MEDLINE | ID: mdl-8782697

ABSTRACT

Phylogenetic relationships among species of the genus Ureaplasma were elucidated by analyzing 16S rRNA sequence information. The 16S rRNA genes of six strains of the mammalian Ureaplasma species were amplified by PCR and were sequenced directly by a primer walking method. The phylogenetic tree based on the nucleotide sequence of the 16S rRNA genes corresponded to the evolutionary history of the host animal species.


Subject(s)
RNA, Ribosomal, 16S/genetics , Ureaplasma/classification , Animals , Base Sequence , DNA, Bacterial , Genes, Bacterial , Mammals/microbiology , Molecular Sequence Data , Phylogeny , RNA, Bacterial , Sequence Homology, Nucleic Acid , Ureaplasma/genetics
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