Molecular Characterization of Mycoplasma pneumoniae Isolates in the United States from 2012 to 2018.

Mycoplasma pneumoniae is a major cause of community-acquired pneumonia. There are limited data in the United States on the molecular epidemiological characteristics of M. pneumoniae We collected 446 M. pneumoniae-positive specimens from 9 states between August 2012 and October 2018. Culture, antimicrobial susceptibility testing, P1 subtyping, and multilocus VNTR (variable-number tandem repeats) analysis (MLVA) were performed to characterize the isolates. Macrolide-resistant M. pneumoniae (MRMp) was detected in 37 (8.3%) specimens. P1 subtype 2 (P1-2) was the predominant P1 subtype (59.8%). P1 subtype distribution did not change significantly chronologically or geographically. The macrolide resistance rate in P1 subtype 1 (P1-1) samples was significantly higher than that in P1-2 (12.9% versus 5.5%). Six P1-2 variants were identified, including two novel types, and variant 2c was predominant (64.6%). P1-2 variants were distributed significantly differently among geographic regions. Classical P1-2 was more frequent in lower respiratory tract specimens and had longer p1 trinucleotide repeats. Classical P1-2 was most common in MRMp (35.7%), while variant 2c was most common in macrolide-susceptible M. pneumoniae (67.5%). Fifteen MLVA types were identified; 3-5-6-2 (41.7%), 4-5-7-2 (35.3%), and 3-6-6-2 (16.6%) were the major types, and four MLVA clusters were delineated. The distribution of MLVA types varied significantly over time and geographic location. The predominant MLVA type switched from 4-5-7-2 to 3-5-6-2 in 2015. MLVA type was associated with P1 subtypes and P1-2 variant types but not with macrolide resistance. To investigate the M. pneumoniae genotype shift and its impact on clinical presentations, additional surveillance programs targeting more diverse populations and prolonged sampling times are required.

Cr2Te3 Thin Films for Integration in Magnetic Topological Insulator Heterostructures.

Chromium telluride compounds are promising ferromagnets for proximity coupling to magnetic topological insulators (MTIs) of the Cr-doped (Bi,Sb)2(Se,Te)3 class of materials as they share the same elements, thus simplifying thin film growth, as well as due to their compatible crystal structure. Recently, it has been demonstrated that high quality (001)-oriented Cr2Te3 thin films with perpendicular magnetic anisotropy can be grown on c-plane sapphire substrate. Here, we present a magnetic and soft x-ray absorption spectroscopy study of the chemical and magnetic properties of Cr2Te3 thin films. X-ray magnetic circular dichroism (XMCD) measured at the Cr L2,3 edges gives information about the local electronic and magnetic structure of the Cr ions. We further demonstrate the overgrowth of Cr2Te3 (001) thin films by high-quality Cr-doped Sb2Te3 films. The magnetic properties of the layers have been characterized and our results provide a starting point for refining the physical models of the complex magnetic ordering in Cr2Te3 thin films, and their integration into advanced MTI heterostructures for quantum device applications.

Crossover from lattice to plasmonic polarons of a spin-polarised electron gas in ferromagnetic EuO.

Strong many-body interactions in solids yield a host of fascinating and potentially useful physical properties. Here, from angle-resolved photoemission experiments and ab initio many-body calculations, we demonstrate how a strong coupling of conduction electrons with collective plasmon excitations of their own Fermi sea leads to the formation of plasmonic polarons in the doped ferromagnetic semiconductor EuO. We observe how these exhibit a significant tunability with charge carrier doping, leading to a polaronic liquid that is qualitatively distinct from its more conventional lattice-dominated analogue. Our study thus suggests powerful opportunities for tailoring quantum many-body interactions in solids via dilute charge carrier doping.

Genotypic characterization of Ureaplasma species by pulsed field gel electrophoresis.

We describe the first use of pulsed field gel electrophoresis to genotype human Ureaplasma species. This technique can distinguish between U. urealyticum and U. parvum, differentiate most of the 14 serovars from one another, and identify differences among clinical isolates of the same serovar.

Midtrimester vaginal Mycoplasma genitalium in women with subsequent spontaneous preterm birth.

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.

Comparative potency of gemifloxacin, new quinolones, macrolides, tetracycline and clindamycin against Mycoplasma spp.

This study determined the comparative in vitro potency of the new investigative quinolone gemifloxacin (SB-265805) using low-passaged clinical isolates and type strains of mycoplasma commonly found in the human respiratory and urogenital tracts. Organisms studied were Mycoplasma pneumoniae, Mycoplasma hominis, Mycoplasma fermentans, Mycoplasma genitalium, Mycoplasma penetrans and Ureaplasma urealyticum, obtained from different geographical regions. Comparator drugs were levofloxacin, trovafloxacin, grepafloxacin, azithromycin, clarithromycin, tetracycline and clindamycin. MICs were determined using a microbroth dilution method. The overall range of MICs of gemifloxacin was < or =0.008-0.125 mg/L for different Mycoplasma spp. and < or =0.008-0.5 mg/L for Ureaplasma spp. Depending on the species tested, gemifloxacin showed variable results when compared with the macrolides. However, gemifloxacin was as potent as, or more potent than, tetracycline, clindamycin and the other quinolones investigated.

Mycoplasma pneumoniae: a frequent cause of pneumonia among U.S. Marines in southern California.

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.

Evaluation of the Etest for detection of tetracycline resistance in Mycoplasma hominis.

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.

Serum concentrations of erythromycin after intravenous infusion in preterm neonates treated for Ureaplasma urealyticum infection.

Erythromycin is receiving renewed attention as an alternative for treatment of neonatal infections caused by Ureaplasma urealyticum because of recently proved abilities of this organism to produce systemic disease in this population. Although erythromycin has been used clinically for almost 40 years, very little is known about its activity in the preterm neonate. Fourteen neonates, birth weights < or = 1500 g and < or = 15 days of age, from whom U. urealyticum was isolated from the lower respiratory tract were randomized to receive erythromycin lactobionate either 25 or 40 mg/kg/day in four divided doses at 6-hour intervals scheduled for a total of 10 days. Blood samples collected at multiple time points after initial and steady state doses were assayed for erythromycin by liquid chromatography. Minimal inhibitory concentrations (MICs) of erythromycin for the U. urealyticum isolates were determined. MICs ranged from 0.031 to 2 micrograms/ml; MIC90 = 2 micrograms/ml. Serum erythromycin concentrations met or exceeded most MICs, with peak values of 3.05 to 3.69 and 1.92 to 2.9 micrograms/ml for the 40- and 25-mg/kg/day dosage groups, respectively. Pharmacokinetic parameters were calculated after the initial dose and at steady state for both dosage groups and compared. No adverse effects thought to be related to administration of erythromycin were observed. These preliminary findings showed that erythromycin is well-tolerated, has favorable pharmacokinetic activity in the preterm neonate and should be further investigated for treatment of ureaplasmal infections.

Septic arthritis and bacteremia due to Mycoplasma resistant to antimicrobial therapy in a patient with systemic lupus erythematosus.

We report a case of septic arthritis caused by Mycoplasma hominis in a patient with systemic lupus erythematosus. The infection started as monarthritis but spread to at least four joints despite apparently suitable therapy with various antimicrobial agents, including doxycycline, clindamycin, and ciprofloxacin; subsequent bacteremia was documented. Control was ultimately achieved with use of the experimental fluoroquinolone temafloxacin in combination with doxycycline administration, arthroscopic drainage of a persistently infected joint, several intravenous infusions of immunoglobulins (which led to increases in levels of antibodies specific to M. hominis), and discontinuation of corticosteroid therapy. Antimicrobial susceptibility testing of various mycoplasmal isolates showed the presence of the tetM gene, disparity between susceptibility to tetracycline and doxycycline, and increasing resistance to most antimicrobial agents used (including to fluoroquinolones before clinical use), although the patient ultimately had a favorable clinical response to treatment with combined modalities.

Rapid detection of tetM in Mycoplasma hominis and Ureaplasma urealyticum by PCR: tetM confers resistance to tetracycline but not necessarily to doxycycline.

Tetracycline resistance in Mycoplasma hominis and Ureaplasma urealyticum has been associated with the tetM determinant and has recently been increasing in incidence. We report here a rapid method for detection of the tetM determinant based on the use of the polymerase chain reaction (PCR) to amplify a 397-bp DNA fragment from the tetM gene and verification of specificity using the restriction enzyme TaqI. Analysis of 42 U. urealyticum and 49 M. hominis isolates indicates that the PCR method may be clinically useful for determination of tetracycline sensitivity, as tetM is presently the only known determinant associated with tetracycline resistance in these two organisms. All of the tetM-positive M. hominis isolates were sensitive to doxycycline, indicating that tetM does not necessarily confer resistance to this antibiotic.

In vitro susceptibilities of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum to sparfloxacin and PD 127391.

The in vitro activities of two investigational quinolones, sparfloxacin (previously designated AT 4140) and PD 127391, were determined for 30 strains each of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum and compared with those of ciprofloxacin, tetracycline, clindamycin, and erythromycin. Erythromycin was the most active compound against M. pneumoniae (maximum MIC, less than 0.008 microgram/ml). PD 127391 (MICs, less than 0.008 to 0.031 microgram/ml), sparfloxacin (MICs, less than 0.008 to 0.25 microgram/ml), clindamycin (MICs, less than 0.008 to 0.5 microgram/ml), and tetracycline (MICs, 0.063 to 0.25 microgram/ml) were superior to ciprofloxacin (MICs, 0.5 to 2 microgram/ml). Sparfloxacin and PD 127391 were active against M. hominis (MICs, less than 0.008 to 0.031 microgram/ml for each) at concentrations comparable to those of clindamycin (MICs, less than 0.008 to 0.063 microgram/ml) and at concentrations lower than those of ciprofloxacin (MICs, 0.125 to 0.5 microgram/ml). As expected, M. hominis was resistant to erythromycin (MICs, 32 to greater than or equal to 256 micrograms/ml). For U. urealyticum, PD 127391 (MICs, 0.031 to 0.5 microgram/ml) and sparfloxacin (MICs, 0.063 to 1 microgram/ml) were superior to erythromycin (MICs, 0.25 to 4 micrograms/ml), ciprofloxacin (MICs, 0.5 to 8 micrograms/ml), and clindamycin (MICs, 0.25 to 64 micrograms/ml. Both new quinolones were equally active against tetracycline-susceptible as well as resistant strains of M. hominis and U. urealyticum. The possible influence of medium components and/or pH on MICs was evaluated by testing a Staphylococcus aureus reference strain with each antibiotic in SP-4 broth and 10-B broth and comparing the results with published MICs for this strain. MICs determined in 10-B broth for erythromycin were affected most. This study shows that the activities of sparfloxacin and PD 127391 are similar to one another and comparable or superior to those of other drugs used to treat mycoplasmal infections. The MICs of both new quinolones were consistently 2 to several dilutions lower than those of ciprofloxacin for each species.

Comparison of agar versus broth dilution techniques for determining antibiotic susceptibilities of Ureaplasma urealyticum.

We determined minimal inhibitory concentrations (MIC) for tetracycline and erythromycin for 72 clinical isolates using broth and agar dilution methods. Erythromycin MIC ranges were less than 0.125-8 micrograms/ml and 1-64 micrograms/ml in broth and agar, respectively. The erythromycin MIC50 and MIC90 as determined by broth were two dilutions (fourfold) lower than those for agar. Tetracycline MIC ranges in broth and agar were less than 0.125-greater than 64 and 0.25-greater than 64 micrograms/ml, respectively. The tetracycline broth MIC50 was one dilution lower than that for agar. The tetracycline broth MIC90 was 64 micrograms/ml and that for agar was greater than 64 micrograms/ml. Of the strains tested, 98.6% using broth were susceptible or moderately susceptible to erythromycin as compared with 75% using agar, representing a significant difference (p less than 0.001). For tetracycline, 80.6% of strains were susceptible or moderately susceptible using broth and 73.6% using agar. MICs were determined by agar dilution after 72 and 96 hr of incubation in 32 strains. There was an increase in the erythromycin MIC by one dilution in 16 strains and two dilutions in one strain with the longer incubation. The tetracycline MIC increased by one dilution in nine strains between readings. Broth MICs were reproducible with one dilution for both drugs in 10 of 12 strains tested twice. Agar MICs were reproducible within a maximum of two dilutions (fourfold). Different interpretations of susceptibility versus resistance may be made depending on which assay is utilized, thus influencing conclusions regarding spectrum of activity of investigational drugs as well as treatment options. The technique employed should always be considered whenever apparently differing drug susceptibility patterns are reported.

Efficacy of clarithromycin against Mycoplasma pneumoniae.

The in-vitro susceptibility of Mycoplasma pneumoniae to clarithromycin, a new macrolide, was compared with that to erythromycin. A broth microdilution assay was used to evaluate 30 clinical isolates collected over a 15 year period from four countries (United States, Australia, Denmark and Japan). Clarithromycin inhibited the growth of all M. pneumoniae strains at low concentrations (less than or equal to 0.008 mg/l) similarly to erythromycin (less than or equal to 0.008 mg/l). In 120 outpatients with radiographically confirmed community acquired pneumonia (mean age 46.2 years), M. pneumoniae was detected culturally and/or serologically by ELISA and immunoblotting in 13% of patients, thus confirming the continued importance of this organism as a respiratory pathogen. M. pneumoniae isolates from these patients were shown to be equally susceptible to clarithromycin (less than or equal to 0.008 mg/l) and erythromycin (less than or equal to 0.008 mg/l). Both clarithromycin and erythromycin were effective in the treatment of community-acquired pneumonia. There were no statistically significant or clinically important differences between the two treatment groups with respect to clinical or radiographic resolution of disease or improvement in signs and symptoms. While the number of clinically evaluable patients with M. pneumoniae infections was small, both macrolides seemed equally effective.

Mycoplasmal infections of cerebrospinal fluid in newborn infants from a community hospital population.

Mycoplasma hominis or Ureaplasma urealyticum have previously been isolated from cerebrospinal fluid (CSF) in 13 of 100 newborn infants tested from a high risk university hospital population where the mothers were of predominantly lower income and socioeconomic status and had often received little or no prenatal care. We sought to determine whether such infections occur in neonates born to women cared for mainly through private obstetric practices and who delivered in 4 suburban community hospitals. CSF cultures were done in 318 infants during an 8-month period. M. hominis was isolated from 9 and U. urealyticum from 5 CSF cultures. Four infants infected with U. urealyticum and 3 infected with M. hominis were born at term. One infant infected with U. urealyticum had a birth weight of less than 1000 g. In 5 infants clearance of the infecting organism was documented without specific treatment. Twelve infants had good perinatal outcomes regardless of treatment and 2 died. One death in a 2240-g infant infected with M. hominis was associated with Haemophilus influenzae sepsis and pneumonia. The other death occurred 3 days after birth in a 630-g infant infected with U. urealyticum who had evidence of meningitis and intraventricular hemorrhage. Results of this study suggest that mycoplasmas are common causes of neonatal CSF infections, not only in high risk populations, but also in the general population.

Comparative susceptibility of Mycoplasma pneumoniae to erythromycin, ciprofloxacin, and lomefloxacin.

Lomefloxacin was found to be comparable to ciprofloxacin in its ability to inhibit the in vitro growth of Mycoplasma pneumoniae (MIC range 2-8 mcg/ml), but it was significantly less active than erythromycin. Although 30 different strains from widely differing geographic areas and isolation time periods were examined, no macrolide-resistant strains were observed.

Ureaplasma urealyticum pneumonia: experimental production and demonstration of age-related susceptibility.

Two different strains of Ureaplasma urealyticum isolated in pure culture from the lungs of newborn human infants were shown to produce an acute, self-limiting, interstitial pneumonia in newborn C3H/HeN and C57BL/6 mice that were free of other respiratory pathogens. Lesion severity peaked 3 to 6 days following intranasal inoculation of ureaplasmas and was resolved by 12 days. Rhinitis and otitis also occurred but did so less frequently than pneumonia. Organisms were localized within the alveoli in areas of inflammation. In comparison with newborn mice, 14-day-old mice were less susceptible to either colonization or disease.

Protein antigens of genital mycoplasmas.

Surface structures of the genital mycoplasmas Ureaplasma urealyticum and Mycoplasma hominis that are important in the human immune response and pathogenesis of disease are relatively poorly defined. In this study, an unusual antigen complex of U. urealyticum consisting of multiple bands forming a "ladder" pattern after electrophoretic separation was noted. It is similar to the variable V-1 surface antigen of Mycoplasma pulmonis. Data on U. urealyticum are only preliminary, but the ureaplasma antigen, if it proves to be analogous to V-1, may provide the antigenic determinants for distinguishing among serovars or serogroups and correlating them with pathogenicity. Surface proteins of M. hominis were identified with use of 125I surface labeling, [35S]methionine metabolic labeling, and immunoadsorption of rabbit antiserum. Comparison of M. hominis reference strains PG-21 and 4195 showed little homology between surface proteins, although with metabolic labeling they appeared essentially identical. Immunoblotting with patients' sera, using PG-21 as antigen, showed that most reactions were directed to surface proteins and that a 102K antigen (MH1) was recognized by 94% of the sera. MH1 was one of the few surface proteins of PG-21 that appeared to have counterparts in the other six reference strains, making MH1 a prime candidate for reliable and specific detection of M. hominis infection.