Ureaplasma Urealyticum Serovars Research Articles

Detection of Ureaplasma spp. serovars in genital tract of infertile males.

BackgroundThe colonization of Ureaplasma species in genital tract is related with male infertility. However, it has been postulated based upon limited study that virulence is related to serotype specificity. The aim of this study was to determine the distribution of Ureaplasma serovars in genital tract of infertile males and analyze their role in male infertility.MethodsA total of 358 urethral swabs samples were obtained from infertile males. The culture of Ureaplasma species were performed using a commercially available Mycoplasma IST 2 kit. Serovars were determined by real‐time polymerase chain reaction (real‐time PCR).ResultsA total of 92 (25.7%) infertile males were positive for Ureaplasma spp; among them, Ureaplasma parvum (UPA) was detected in 73 (79.3%) isolates, and Ureaplasma urealyticum (UUR) was detected in 19 (20.7%) isolates. Serovars 1, 6, or in combination accounted for 63.0% (46/73) of UPA isolates. Serovar 9 (alone and in combination of other serovars) was the most common serovar in UUR (47.4%, 9/19). Multiple serovars were detected in 21 (22.8%) isolates, and serovars 4, 5, 7, and 12 were not detected in any sample.ConclusionThe distribution of 14 Ureaplasma serovars in genital tract of infertile males was identified for the first time by real‐time PCR assay. UPA serovars 1 and 6, and UUR serovar 9 are the most common serovars colonization in urogenital tract of infertile males.

Ureaplasma spp. lipid-associated membrane proteins induce human monocyte U937 cell cycle arrest through p53-independent p21 pathway

Ureaplasma spp. are known to be associated with human genitourinary tract diseases and perinatal diseases and Ureaplasma spp. Lipid-associated membrane proteins (LAMPs) play important roles in their related diseases. However, the exact mechanism underlying pathogenesis of Ureaplasma spp. LAMPs is largely unknown. In this study, we explored the pathogenic mechanisms of Ureaplasma spp. LAMPs by elucidating their role in modulating the cell cycle and related signaling pathways in human monocytic cell U937, which is highly related to the inflammatory and protective effect in infectious diseases. We utilized the two ATCC reference strains (Ureaplasma parvum serovar 3 str. ATCC 27,815 (UPA3) and Ureaplasma urealyticum serovar 8 str. ATCC 27,618 (UUR8)) and nine clinical isolates which including both UPA and UUR to study the effects of Ureaplasma spp. LAMPs on U937 in vitro. We found that LAMPs derived from UUR8 and both UPA and UUR of clinical strains markedly inhibited the cell proliferation, while UPA3 LAMPs suppressed slightly. Besides, the result of flow cytometry analysis indicated all the Ureaplasma spp. LAMPs could arrest U937 cells in G1 phase. Next, we found that the cell cycle arrest was associated with increased levels of p53 and p21, and a concomitant decrease in the levels of CDK2, CDK4, CDK6 and cyclin E1 at both transcriptional and translational levels after treatment with LAMPs derived from UUR8 or clinical strains, while only cyclin E1 was down-regulated after treatment with UPA3 LAMPs. Further study showed that p53 down-regulation had almost no effect on the distribution of cell cycle and the expression of p21. In conclusion, this study demonstrated that LAMPs derived from UUR8 and clinical strains could inhibit the proliferation of U937 cells by inducing G1 cell cycle arrest through increasing the p21 expression in a p53-independent manner, while UPA3 LAMPs could induce the cell cycle arrest slightly. Our study could contribute to the understanding of Ureaplasma spp. pathogenesis, which has potential value for the treatment of Ureaplasma spp. infections.

Novel insights into neuroinflammation: bacterial lipopolysaccharide, tumor necrosis factor \u03b1, and Ureaplasma species differentially modulate atypical chemokine receptor 3 responses in human brain microvascular endothelial cells

BackgroundAtypical chemokine receptor 3 (ACKR3, synonym CXCR7) is increasingly considered relevant in neuroinflammatory conditions, in which its upregulation contributes to compromised endothelial barrier function and may ultimately allow inflammatory brain injury. While an impact of ACKR3 has been recognized in several neurological autoimmune diseases, neuroinflammation may also result from infectious agents, including Ureaplasma species (spp.). Although commonly regarded as commensals of the adult urogenital tract, Ureaplasma spp. may cause invasive infections in immunocompromised adults as well as in neonates and appear to be relevant pathogens in neonatal meningitis. Nonetheless, clinical and in vitro data on Ureaplasma-induced inflammation are scarce.MethodsWe established a cell culture model of Ureaplasma meningitis, aiming to analyze ACKR3 variances as a possible pathomechanism in Ureaplasma-associated neuroinflammation. Non-immortalized human brain microvascular endothelial cells (HBMEC) were exposed to bacterial lipopolysaccharide (LPS) or tumor necrosis factor-α (TNF-α), and native as well as LPS-primed HBMEC were cultured with Ureaplasma urealyticum serovar 8 (Uu8) and U. parvum serovar 3 (Up3). ACKR3 responses were assessed via qRT-PCR, RNA sequencing, flow cytometry, and immunocytochemistry.ResultsLPS, TNF-α, and Ureaplasma spp. influenced ACKR3 expression in HBMEC. LPS and TNF-α significantly induced ACKR3 mRNA expression (p < 0.001, vs. control), whereas Ureaplasma spp. enhanced ACKR3 protein expression in HBMEC (p < 0.01, vs. broth control). Co-stimulation with LPS and either Ureaplasma isolate intensified ACKR3 responses (p < 0.05, vs. LPS). Furthermore, stimulation wielded a differential influence on the receptor’s ligands.ConclusionsWe introduce an in vitro model of Ureaplasma meningitis. We are able to demonstrate a pro-inflammatory capacity of Ureaplasma spp. in native and, even more so, in LPS-primed HBMEC, underlining their clinical relevance particularly in a setting of co-infection. Furthermore, our data may indicate a novel role for ACKR3, with an impact not limited to auto-inflammatory diseases, but extending to infection-related neuroinflammation as well. AKCR3-induced blood-brain barrier breakdown might constitute a potential common pathomechanism.

Ureaplasma isolates differentially modulate growth factors and cell adhesion molecules in human neonatal and adult monocytes

Generally regarded as commensal bacteria, the pathogenicity of Ureaplasma has often been considered low. Controversy remains concerning the clinical relevance of Ureaplasma infection in the pathogenesis of inflammation-related morbidities. Recently, we demonstrated Ureaplasma-driven pro-inflammatory cytokine responses in human monocytes in vitro. We hypothesized that Ureaplasma may induce further inflammatory mediators. Using qRT-PCR and multi-analyte immunoassay, we assessed the expression of granulocyte-colony stimulating factor (G-CSF), vascular endothelial growth factor (VEGF), intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) in term neonatal and adult monocytes exposed to Ureaplasma urealyticum serovar 8 (Uu8) and U. parvum serovar 3 (Up3). Ureaplasma significantly induced VEGF mRNA in neonatal (Up3: p < 0.05, versus broth control) and adult monocytes (Uu8: p < 0.05) as well as ICAM-1 mRNA in neonatal cells (p < 0.05 each). As far as protein expression was concerned, Up3 stimulated VEGF release in both monocyte subsets (p < 0.01) and enhanced secretion of ICAM-1 protein in neonatal monocytes (p < 0.05). In adult cells, ICAM-1 protein release was increased upon exposure to both isolates (Uu8: p < 0.05, Up3: p < 0.01). Ureaplasma-induced responses did not significantly differ from corresponding levels mediated by E. coli lipopolysaccharide (LPS). The stimulatory effects were dose-dependent. Ureaplasma infection, on the contrary, did not affect G-CSF and VCAM-1 expression. Of note, co-infection of LPS-primed neonatal monocytes with Ureaplasma enhanced LPS-induced ICAM-1 release (Uu8: p < 0.05). Our results confirm Ureaplasma-driven pro-inflammatory activation of human monocytes in vitro, demonstrating a differential modulation of growth factors and cell adhesion molecules, that might promote unbalanced monocyte responses and adverse immunomodulation.

Ureaplasma parvum Prosthetic Joint Infection Detected by PCR

We describe the first reported case of Ureaplasma parvum prosthetic joint infection (PJI) detected by PCR. Ureaplasma species do not possess a cell wall and are usually associated with colonization and infection of mucosal surfaces (not prosthetic material). U. parvum is a relatively new species name for certain serovars of Ureaplasma urealyticum, and PCR is useful for species determination. Our patient presented with late infection of his right total knee arthroplasty. Intraoperative fluid and tissue cultures and pre- and postoperative synovial fluid cultures were all negative. To discern the pathogen, we employed PCR coupled with electrospray ionization mass spectrometry (PCR/ESI-MS). Our patient's failure to respond to empirical antimicrobial treatment and our previous experience with PCR/ESI-MS in culture-negative cases of infection prompted us to use this approach over other diagnostic modalities. PCR/ESI-MS detected U. parvum in all samples. U. parvum-specific PCR testing was performed on all synovial fluid samples to confirm the U. parvum detection.

Serological investigation ofUreaplasma urealyticumin Korean preterm infants

PurposeUreaplasma colonization is related with perinatal complications in preterm infants. Little is known about the difference in virulence among various Ureaplasma urealyticum serovars. The aim of this study was to determine U. urealyticum serovars of preterm infants in order to assess whether any of the serovars were associated with bronchopulmonary dysplasia (BPD).MethodsThree hundred forty-four preterm infants with a gestational age less than 34 weeks admitted to Gangnam Severance Hospital neonatal intensive care unit from July 2011 to December 2012 were included in this study. Tracheal and gastric aspirations were conducted on infants to confirm Ureaplasma colonization. Ureaplasma colonization was confirmed in 9% of infants, of these, serovars were determined by real-time polymerase chain reaction.ResultsA total of 31 infants (gestational age, 29.3±3.1 weeks; birth weight, 1,170±790 g) were U. urealyticum positive. The Ureaplasma positive group treated for more days with oxygen and ventilation than the negative group (P<0.05). Histologic chorioamnionitis and moderate to severe BPD were more frequent in the Ureaplasma positive group than in the negative group (P<0.05). U. urealyticum isolates were either found to be a mixture of multiple serovars (32%), serovar 9 alone or combined with other serovars (39%), serovar 11 (26%), 2 (13%), 8 (10%), 10 (13%), and 13 (25%). No individual serovars were significantly associated with moderate to severe BPD and chorioamnionitis.ConclusionThis is the first study to describe the distribution of U. urealyticum serovars from Korean preterm infants. Ureaplasma-colonized infants showed higher incidence of BPD and chorioamnionitis.

Frequency of Ureaplasma Serovars in Respiratory Secretions of Preterm Infants at Risk for Bronchopulmonary Dysplasia

Ureaplasma respiratory tract colonization is associated with bronchopulmonary dysplasia (BPD) in preterm infants. Whether the 4 Ureaplasma parvum and 10 Ureaplasma urealyticum serovars differ in virulence is unknown. This study was conducted to determine the distribution of Ureaplasma serovars in respiratory secretions of a prospective cohort of preterm infants and to assess whether any of the serovars are associated with BPD. Serial endotracheal and/or nasopharyngeal aspirates were obtained for Ureaplasma culture and PCR from 136 infants of gestational age <33 weeks. All positive samples were speciated and serovars were determined by real-time PCR. A total of 51 (37.5%) infants were Ureaplasma-positive one or more times during the first month of life. Respiratory colonization was inversely related to gestational age. Sixty-five percent of infants <26 weeks compared with 31% infants ≥ 26 weeks were culture or PCR positive. U. parvum was more common (N = 32, 63%) than U. urealyticum (N = 17, 33%); both species were present in 2 samples. Serovars 3 and 6 alone and in combination accounted for 96% U. parvum isolates. U. urealyticum isolates were commonly a mixture of multiple serovars, with serovar 11 alone or combined with other serovars (10/17, 59%) being the most common serovar. No individual species or serovars or serovar mixtures were associated with moderate-to-severe BPD. U. parvum serovars 3 and 6 and U. urealyticum serovar 11 were the most common serovars detected in respiratory samples from a prospective cohort of preterm infants.

Comparison of full gyrA, gyrB, parC and parE gene sequences between all Ureaplasma parvum and Ureaplasma urealyticum serovars to separate true fluoroquinolone antibiotic resistance mutations from non-resistance polymorphism

To determine the role of amino acid substitutions in Ureaplasma GyrA, GyrB, ParC and ParE proteins in mediating fluoroquinolone resistance. Nucleic acid sequences from gyrA, gyrB, parC and parE genes from all 14 Ureaplasma serovars were aligned. Full genome sequences for serovars 1, 3-7, 9 and 11-14 were available from the National Center for Biotechnology Information database and we sequenced the full topoisomerase genes from ciprofloxacin-susceptible reference strains of serovars 2, 8 and 10. Phylogenetic trees were constructed to analyse nucleotide sequence similarity. Deduced amino acid sequences were compared with all 33 previously reported fluoroquinolone-resistant strains to clarify true fluoroquinolone-resistance-associated substitutions. Non-resistance-associated polymorphisms were identified in GyrA (39), GyrB (26), ParC (107) and ParE (34) proteins. Phylogenetic analysis demonstrated species clustering for all genes, except parE in which serovars 4, 12, 10 and 13 formed a separate cluster more similar to Ureaplasma parvum than the remaining Ureaplasma urealyticum serovars. Examination of all previously reported fluoroquinolone-resistant strains found that one-third of identified residue substitutions could be attributed to normal species polymorphism; therefore, the mechanism of resistance for these strains is still undetermined. In particular, Glu or Asp at position 112 in GyrA and Ala or Thr at 125/136 in ParC were substitutions identified when U. urealyticum strain sequences were previously aligned with the published serovar 3 genome sequence. Combining analysis of the recently available Ureaplasma genomes with sequences from the additional serovars has enabled us to clarify which substitutions found by previous investigators could potentially be responsible for fluoroquinolone resistance.

Differences in biofilm development and antibiotic susceptibility among clinical Ureaplasma urealyticum and Ureaplasma parvum isolates

The aim of this work was to study the ability of clinical isolates of Ureaplasma spp. to form biofilms in vitro and to compare the antibiotic susceptibility of sessile cells and their planktonic counterparts. A total of nine Ureaplasma spp. isolates recovered from unrelated male patients diagnosed with urethritis or chronic prostatitis and two isolates isolated from the urine of two healthy volunteers were included. Ureaplasma species identification was performed by 16S rDNA gene amplification and sequencing. Conventional antibiotic susceptibility tests were carried out by the broth microdilution method. Biofilm susceptibility assays were performed following the method proposed by Moskowitz using 10C urea broth medium and confirming bacterial growth by colour shift of the medium. The chi(2) test was applied to analyse the statistical differences between the MIC and the minimal biofilm inhibitory concentration. Isolates were identified as Ureaplasma urealyticum serovar 7 (five isolates), U. urealyticum serovar 13 (four isolates) and Ureaplasma parvum serovar 3 (two isolates). Biofilm formation was observed in 9 out of the 11 strains studied (82%); two isolates of U. urealyticum serovar 13 were non-biofilm formers. Global resistance percentages of planktonic cells compared with sessile cells were different for erythromycin (0% versus 44%, P = 0.02), telithromycin (22% versus 77%, P = 0.02), ciprofloxacin (66% versus 100%), levofloxacin (0% versus 33%) and tetracycline (0% versus 33%). All nine biofilm-forming strains were fully susceptible to clarithromycin in both planktonic and biofilm types of growth. These results indicate that biofilm formation can protect mycoplasma cells from antibiotics and host defences, favouring their persistence in chronically infected or colonized patients while increasing resistance to antimicrobial agents. Therefore, the capacity to form biofilms by Ureaplasma spp. isolates should be considered when antibiotic treatments are required.

Phylogenetic analysis of Ureaplasma urealyticum--support for the establishment of a new species, Ureaplasma parvum.

In this study, the phylogenetic relationships between the two biovars and 14 serovars of Ureaplasma urealyticum were studied using the sequences of four different genes or genetic regions, namely: 16S rRNA genes; 16S-23S rRNA gene spacer regions; urease gene subunits ureA, ureB, partial ureC and adjoining regions upstream of ureA, ureA-ureB spacer and ureB-ureC spacer; the 5'-ends of the multiple-banded antigen (MBA) genes. U. urealyticum genotypes, based on all four genomic sequences, could be clearly separated into two clusters corresponding with currently recognized biovars 1 and 2. Sequences were generally conserved within each biovar. However, there was heterogeneity within the 5'-end regions of the MBA genes of the four serovars of biovar 1; the sequence of serovar 3 was identical with the previously published sequence and differed by only three bases from that of serovar 14; but there were significant differences between the sequences of serovars 3 and 14 and those of serovars 1 and 6. Based on the phylogenetic analysis, support is given to previous recommendations that the two biovars of U. urealyticum be classified as distinct species, namely U. parvum and U. urealyticum for biovars 1 and 2, respectively. In the future, the relationship between the new species and clinical manifestations of ureaplasma infections should be studied.

The phylogeny of Ureaplasma urealyticum based on the mba gene fragment.

Sequencing of mba gene fragments of reference strains of Ureaplasma urealyticum serovars 1, 3, 6, 14 and 8, in addition to 33 clinical U. urealyticum isolates is reported. A phylogenetic tree deduced from alignment of these sequences clearly demonstrates two major clusters (confidence limit 100%), which equate to the parvo and T960 biovars, and five types, which have been designated mba 1, 3, 6, 8 and X. These relationships are supported by bootstrap analysis. Polymorphisms within the mba fragment of types mba 1, 3 and 6 were used to define nine subtypes (mba 1a, 1b, 3a, 3b, 3c, 3d, 3e, 6a and 6b), thus facilitating high resolution typing of U. urealyticum. Inclusion of reference strains for serovars 1, 3, 6 and 8 in the mba typing scheme showed that the results of this analysis are broadly consistent with currently accepted serotyping. In addition, a ure gene fragment from nine of the clinical isolates was amplified and sequenced. Comparisons of the sequences clearly distinguished the two biovars of U. urealyticum; however, this fragment was invariant within the parvo biovar. This study has shown that the mba sequence can reveal the fine details of the relationships between U. urealyticum isolates and also supports the significant evolutionary gap between the two biovars.

Clastogenic effects of different Ureaplasma urealyticum serovars on human chromosomes.

The possibility that Ureaplasma urealyticum might play an important role in human infertility was first raised more than 20 years ago, but this association remains speculative. Considering the hypothesis that the pathogenicity of Ureaplasma urealyticum may depend on its serotypes, the clastogenic effects of different strains of Ureaplasma urealyticum, at concentrations of 10(3) CCU (color changing units)/ml, 10(4) CCU/ml and 10(5) CCU/ml, were evaluated in vitro in short-term cultures of human lymphocytes. Total or partial mitotic inhibition was produced by Ureaplasma urealyticum serotypes 2, 3 and 10 independent of the concentration (10(3) CCU/ml, 10(4) CCU/ml or 10(5) CCU/ml) of the microorganisms employed. In contrast, the clastogenic effects observed with serotypes 1, 7 and 12 varied according to the concentration employed in the test. Mitotic alterations were observed in Ureaplasma urealyticum serotypes 5, 6, 7, 8, 9, 11 and 12. Chromatid gaps (53.0%) and chromatid breaks (13.9%) were the most frequent types of alterations observed. The results of this in vitro assay demonstrated that the clastogenic effects varied with the Ureaplasma urealyticum serotypes evaluated.

Two Genomic Clusters among 14 Serovars of Ureaplasma urealyticum

Fourteen established serovars of Ureaplasma urealyticum were examined by restriction endonuclease cleavage patterns of genomic DNA, Southern blot hybridization and DNA-DNA hybridization. The restriction endonuclease cleavage patterns and the Southern blot hybridization both revealed that the 14 serovars are divided into two clusters. The DNA-DNA hybridization data further confirmed that these two clusters are genetically distinct of each other.

Rheumatoid Arthritis: New Findings on the Failure to Isolate or Detect Mycoplasmas by Multiple Cultivation or Serologic Procedures and a Review of the Literature

Using 12 different and elaborate broth, agar, and cell culture procedures, we failed to isolate mycoplasmas, ureaplasmas, spiroplasmas, or chlamydiae from the synovial fluid of 10 patients with rheumatoid arthritis (RA) and from six patients with non-rheumatoid arthritis (NRA). In addition, sera from 35 patients with RA and 12 patients with NRA also were examined. Although some of the sera had moderately high titers of metabolism-inhibiting antibody to some of the 10 human Mycoplasma species, especially to the common respiratory pathogen Mycoplasma pneumoniae, and to some of the eight Ureaplasma urealyticum serovars, especially serovars V and VII, there were no significant differences between titers of these antibodies in the two groups of patients. Among RA patients serum antibody titers to M. pneumoniae were 1:32 in five and 1:16 in eight; two patients had higher synovial fluid titers (1:16) than serum titers (1:4). The geometric mean titer (GMT) of antibody to serovar V in synovial fluid was higher in RA patients than in NRA patients, but the difference did not reach significance (P = .056). Reports on the possible role of infectious agents in the pathogenesis of rheumatoid arthritis are reviewed.

Variable antigens of Ureaplasma urealyticum containing both serovar-specific and serovar-cross-reactive epitopes

Currently there are 14 recognized serovars of Ureaplasma urealyticum, and it has been postulated that only certain ones may be associated with disease and that lack of serovar-specific antibody may be an important risk factor. Unfortunately, ureaplasma antigens important in the human immune response and disease pathogenesis are poorly defined. By using sera from ureaplasma-infected patients and antiureaplasma monoclonal antibodies, the present study has demonstrated, for serovars 3, 8, and 10, antigens which (i) are species specific, (ii) contain both serovar-specific and cross-reactive epitope(s), (iii) are produced not only in vitro but also in vivo, (iv) undergo a high rate of structural variation in vitro, (v) are present and structurally variable on invasive ureaplasma isolates (i.e., those from placenta, lung, and cerebrospinal fluid), and (vi) are among the predominant antigens recognized during infections in humans. Furthermore, we have shown that monoclonal antibodies to these antigens can inhibit the growth of the organisms in vitro, indicating the potential for these antigens to be important for host defense.

Development and application of monoclonal antibodies for the detection of cell infecting mycoplasmas

Two hybridoma cell lines producing monoclonal antibodies (MAbs) to mycoplasmas were established by fusion of mouse myeloma and spleen cells obtained from mice immunized with whole cell Mycoplasma pneumoniae antigens. The MAbs designated as MYC-4 and MYC-9 bound to a single M. pneumoniae protein band with approximate molecular weights of 150 000. In dot-EIA both MAbs reacted with ten Mycoplasma spp., two serovars of Ureaplasma urealyticum and Acholeplasma laidlawii. The MAbs have great potential to be used as an antibody probe for the rapid screening of cell cultures and hybridomas for mycoplasmal contamination.

Metabolic distinctiveness of ureaplasmas.

Enzymes of the Embden-Meyerhof-Parnas pathway and hexose monophosphate shunt were examined in cytoplasmic extracts of three serovars of Ureaplasma urealyticum. We found no glucose-6-phosphate or 6-phosphogluconate dehydrogenase, hexokinase, phosphoglucose isomerase, aldolase, or lactic dehydrogenase activities. We failed to find cytochrome pigments in extracts and found no significant production of 14CO2 from [U-14C]glucose, nor did we find oxygen-dependent reduced nicotinamide adenine dinucleotide oxidase activity. Lactic acid was found only at trace levels in spent culture fluids. Ureaplasmas are apparently nonfermentative and are unlike all other mollicutes in that they have no detectable oxygen-dependent reduced nicotinamide adenine dinucleotide oxidase activity.

Detection of End Products of the Arginine Dihydrolase Pathway in Both Fermentative and Nonfermentative Mycoplasma Species by Thin-Layer Chromatography

A total of 22 species of the order Mycoplasmatales, representing eight serologically distinct groups, were tested for the presence of the first two enzymes of the arginine dihydrolase pathway by using two-dimensional thin-layer chromatography to separate end products. Both citrulline, produced by arginine deiminase, and ornithine, produced by ornithine carbamoyltransferase, were found in the glycolytic serogroup 1 species Mycoplasma putrefaciens, Mycoplasma capricolum, and Mycoplasma sp. bovine group 7, in Mycoplasma fermentans (serogroup 6), and in the nonfermentative serogroup 7 species Mycoplasma arginini, Mycoplasma hominis, Mycoplasma gallinarum, Mycoplasma gateae, Mycoplasma salivarium, Mycoplasma orale, and Mycoplasma buccale. Citrulline alone was found in one glycolytic species, Mycoplasma gallisepticum (serogroup 4). No end products were found in six other fermentative Mycoplasma species, in three Acholeplasma species, or in two Ureaplasma urealyticum serovars. The use of radiolabeled substrate with representative species showed that citrulline and ornithine were produced from arginine and not derived from the medium.

Immunoglobulin A1 protease activity in strains of Ureaplasma urealyticum.

Thirteen serovars of Ureaplasma urealyticum were analyzed for the ability to cleave human IgA. Strains of all of the serovars tested cleaved IgA 1 in the hinge region of the alpha-chain, resulting in intact Fc and monomeric Fab fragments. IgA 1 protease activity was also observed in concentrated cell-free extracts of spent cultivation medium, indicating that the IgA 1 protease was excreted into the medium during growth of the micro-organisms. Five clinical isolates of U. urealyticum obtained from urine, cervix, vagina, amniotic fluid, and synovial fluid were positive for IgA 1 protease activity. No proteolytic activity was observed against human IgA 2, IgG, or IgM. Strains of Mycoplasma fermentans, M. salivarium and seven serovars of M. hominis were negative for IgA 1 protease activity.

Synthesis of Lipids from Acetate Is Not Characteristic of Acholeplasma or Ureaplasma Species

Previous studies of Acholeplasma species indicated that this genus could be separated from Mycoplasma on the basis of the ability to synthesize lipids from acetate. We report that recently described strains of Acholeplasma florum (strain L1 isolated from a lemon flower and strain GF-1 isolated from a grapefruit flower) and two unclassified Acholeplasma strains (strain J233 from coconut palm and Acholeplasma sp. strain PS-1 from an insect) convert little if any acetate carbon to lipid, in contrast to Acholeplasma sp. strain 0502-CL1 isolated from broccoli and six other established Acholeplasma species of animal origin. Thus, the ability to synthesize lipid from acetate in Edward test medium containing horse serum is not characteristic of all members of the genus Acholeplasma (class Mollicutes), but appears to be a property only of a subgroup in the genus containing the species that are most frequently isolated from animals (Acholeplasma laidlawii, Acholeplasma granularum BTS39, Acholeplasma oculi 19L, Acholeplasma equifetale N93, and probably Acholeplasma axanthum S743 and Acholeplasma modicum PG49). Like other Mycoplasma species which we studied, Mycoplasma bovigenitalium PG11, Mycoplasma hominis PG27, Mycoplasma arginini G230, and four sterol-requiring mycoplasmas of insect or plant origin (Mycoplasma sp. strain MQ3 from an insect, Mycoplasma sp. strain pommier from an apple, Mycoplasma sp. strain melaleuca from a Melaleuca flower, and Mycoplasma sp. strain 831-C4 from lettuce) convert little acetate to lipid. Similarly, eight serovars of Ureaplasma urealyticum convert little if any acetate to lipid.