Intracellular fate of Ureaplasma parvum entrapped by host cellular autophagy.

Genital mycoplasmas, including Ureaplasma spp., are among the smallest human pathogenic bacteria and are associated with preterm birth. Electron microscopic observation of U. parvum showed that these prokaryotes have a regular, spherical shape with a mean diameter of 146 nm. U. parvum was internalized into HeLa cells by clathrin-mediated endocytosis and survived for at least 14 days around the perinuclear region. Intracellular U. parvum reached endosomes in HeLa cells labeled with EEA1, Rab7, and LAMP-1 within 1 to 3 hr. After 3 hr of infection, U. parvum induced the cytosolic accumulation of galectin-3 and was subsequently entrapped by the autophagy marker LC3. However, when using atg7-/- MEF cells, autophagy was inadequate for the complete elimination of U. parvum in HeLa cells. U. parvum also colocalized with the recycling endosome marker Rab11. Furthermore, the exosomes purified from infected HeLa cell culture medium included U. parvum. In these purified exosomes ureaplasma lipoprotein multiple banded antigen, host cellular annexin A2, CD9, and CD63 were detected. This research has successfully shown that Ureaplasma spp. utilize the host cellular membrane compartments possibly to evade the host immune system.

Ureaplasma parvum and Ureaplasma urealyticum are detected in semen after washing before assisted reproductive technology procedures.

OBJECTIVE: To investigate the prevalence of ureaplasmas in semen and washed semen and to explore their effect on semen andrology variables. DESIGN: Prospective study. SETTING: In vitro fertilization (IVF) unit of a private hospital. PATIENT(S): Three hundred forty-three men participating in an assisted reproductive technology (ART) treatment cycle. MAIN OUTCOME MEASURE(S): The prevalence of ureaplasmas in semen and washed semen tested by culture, polymerase chain reaction assays, and indirect immunofluorescent antibody assays. RESULT(S): Ureaplasmas were detected in 73 of 343 (22%) semen samples and 29 of 343 (8.5%) washed semen samples. Ureaplasmas adherent to the surface of spermatozoa were demonstrated by indirect immunofluorescent antibody testing. Ureplasma parvum serovar 6 (36.6%) and U. urealyticum (30%) were the most prevalent isolates in washed semen. A comparison of the semen andrology variables of washed semen ureaplasma positive and negative groups demonstrated a lower proportion of nonmotile sperm in men ureaplasma positive for washed semen. CONCLUSION(S): Ureaplasmas are not always removed from semen by a standard ART washing procedure and can remain adherent to the surface of spermatozoa.

Ureaplasma canigenitalium sp. nov., isolated from dogs.

Ureaplasma strains isolated from dogs (Canis familiaris) were characterized and compared with the type strains of five previously described species of the genus Ureaplasma, Ureaplasma urealyticum (isolated from humans), Ureaplasma diversum (isolated from cattle), Ureaplasma gallorale (isolated from chickens), Ureaplasma cati (isolated from cats), and Ureaplasma felinum (isolated from cats). The canine strains hydrolyzed urea but not arginine or glucose, were membrane bound, lacked a cell wall, passed through 450-nm-pore-size membrane filters, required cholesterol for growth, and formed minute colonies (diameter, 20 to 140 microns) on agar medium. These canine ureaplasma strains have been reported to be members of four serovars. The four serovars of canine strains fell into a single group on the basis of their genomic properties, as determined by DNA-DNA hybridization. On the basis of these findings, we propose that ureaplasmas with these characteristics belong to a new species, Ureaplasma canigenitalium, with strain D6P-C (= ATCC 51252) as the type strain.

Clinical and biological characteristics of Ureaplasma urealyticum induced polyarthritis in a patient with common variable hypogammaglobulinaemia.

Persistent infectious polyarthritis caused by Ureaplasma urealyticum in a patient with common variable hypogammaglobulinaemia is described. The patient developed a symmetrical, destructive polyarthritis and tenosynovitis associated with a markedly depressed synovial fluid glucose concentration and characteristic soft tissue abscesses. The ureaplasma organism developed resistance to multiple antibiotics and persisted for five years. The organism was identified repeatedly in many joints by culture, confirmed by DNA hybridisation, and mycoplasma-like structures were shown in synovial tissues by electron microscopy.

Localization of endogenous activity of phospholipases A and C in Ureaplasma urealyticum.

Endogenous activities of phospholipases A and C in Ureaplasma urealyticum were assayed in cellular fractions of exponential-phase cells. Enzymatic studies indicated that ATPase activity was localized in the plasma membrane fraction and NADH and NADPH dehydrogenase activities were localized in the cytosol fraction. Studies with purified ureaplasma membranes demonstrated that, of three serovars tested, endogenous phospholipase A1, A2, and C activities were localized in the plasma membrane. Very low levels of activity were observed in the cytosol fractions. Phospholipase A2 activity in the plasma membrane was 3- to 5-fold higher than the activity in the lysates and 60- to 300-fold higher than the activity of phospholipase A1. Phospholipase C was localized mainly in the plasma membrane, with 20% found in the cytosol fraction. The levels of activity were comparable among the three serovars. There was a significantly lower level of activity in cells from the stationary growth phase than in the exponential phase. Significant differences were observed in the phospholipase A activities among the U. urealyticum serovars 3, 4, and 8. Phospholipase A2 activity was twofold higher in serovar 8 membranes, and phospholipase A1 activity was twofold higher in serovar 3 membranes. These results demonstrate that endogenous activities of phospholipase A and C are localized primarily in the plasma membrane fraction of U. urealyticum. The specific activities in the membranes of the phospholipases varied among the three serovars. Phospholipase enzymes may function as virulence factors in U. urealyticum and may vary among the serovars.

Hemadsorption by colonies of Ureaplasma urealyticum.

Hemadsorption by colonies of Ureaplasma urealyticum and Mycoplasma pneumoniae differed quantitatively and qualitatively. Using standard methodology, few strains of U. urealyticum hemadsorbed; with a modified method, most strains hemadsorbed, indicating a second type of association. Scanning electron microscopy of tannin-osmium-stained preparations showed guinea pig erythrocytes embedded in ureaplasma colonies and craters left when erythrocytes were dislodged.

Estudio microbiológico y observación morfocitológica de mycoplasma hominis y ureaplasma urealyticum por microscopía electrónica de transmisión / Microbiological study and morphocytological observation of mycoplasma hominis and ureaplasma urealyticum with transmition electron microscopy

Existe un alto grado de interés en el estudio de las especies de Mycoplasmas y su relación con procesos patológicos en el hombre. En este trabajo se realiza un estudio comparativo de 2 técnicas de procesamiento de MET de 2 especies de Mycoplasmas. A partir de medio líquido fue posible observar por MET ambas especies, describiendo sus formas, midiendo su tamaño y haciendo una breve descripción de su ultraestructura

Cervical chlamydia trachomatis and mycoplasmal infections in women with abnormal Papanicolaou smears.

In a series of 2,346 Papanicolaou-stained smears from women undergoing routine gynaecological examination, 39 showed cytomorphological signs of inflammation suggesting Chlamydia trachomatis infection (Papanicolaou class II or III). The 39 smears were studied microbiologically by the direct-immunofluorescence test and cell culture to see whether chlamydial infection correlated with the presence of Mycoplasma hominis and Ureaplasma urealyticum. The results were compared with the cytological and colposcopic findings. C. trachomatis was cultured in 56.41% of the 39 smears, and isolated by the direct-immunofluorescence test in 51.28%. M. hominis was detected in 35.89% and U. urealyticum in 25.54%. Though all three organisms coexisted in 10.25% of the smears, C. trachomatis and M. hominis in 15.38%, C. trachomatis and U. urealyticum in 2.56%, no valid conclusions could be drawn from their association. The study did, however, indicate that vacuolated cells and cells with "bubbly" cytoplasm are common also to other infections seen in PAP-test smears and do not necessarily warrant a diagnosis of C. trachomatis, but that Gupta-type intracellular inclusion bodies do.

In vitro exposure of bovine morulae to Ureaplasma diversum.

Ureaplasma diversum has been associated with infertility in the cow experimentally and in naturally occurring cases. However, the pathogenic mechanism is undetermined. The purpose of this study was to determine whether ureaplasmas are pathogenic for bovine morulae in vitro. Twenty-one morulae were recovered from three superovulated, mature, Holstein cows six or seven days postestrus. The embryos were divided into three groups (A,B,C) and incubated for 16 hours at 37 degrees C in humidified air with 10% CO2. Group A was incubated in embryo culture medium alone, Group B was incubated in culture medium with sterile ureaplasma broth added and Group C was incubated in culture medium containing 1.7 X 10(6) colony forming units Ureaplasma diversum strain 2312. After incubation, the morulae were examined using an electron microscope. Structures morphologically identical to U. diversum were present on the outer surface of the zonae pellucidae of all the morulae exposed to the organism and none were present on the unexposed control embryos. No other morphological differences were observed in either the ureaplasma-exposed embryos or the two groups of control embryos. Ureaplasma diversum was isolated from three of the five embryos incubated in culture medium with sterile ureaplasma broth added. These three embryos were recovered from one donor cow which cultured positive for U. diversum from the vulva and flush fluid. This finding suggests that the contaminating organisms entered the embryo culture wells either in the embryo collection medium or attached to the embryos.(ABSTRACT TRUNCATED AT 250 WORDS)

Ultrastructural changes in mollicutes induced by the peptide antibiotic herbicolin A.

Electron microscopy of negatively stained mycoplasma, ureaplasma, and acholeplasma cells showed ultrastructural changes after 10 min of treatment of the organisms with the peptide antibiotic herbicolin A in concentrations ranging from 10 micrograms/ml for Mycoplasma capricolum to 600 micrograms/ml for Ureaplasma urealyticum. The morphological changes were shown to be reversible at low concentrations of the antibiotic but irreversible at high concentrations.

Colony morphology, ultrastructure and morphogenesis in Mycoplasma hominis, Acholeplasma laidlawii and Ureaplasma urealyticum.

Colonies of Mycoplasma hominis, Acholeplasma laidlawii (three strains) and Ureaplasma urealyticum were examined by light and electron microscopy and their characteristic morphology, ultrastructure and morphogenesis are described. Mycoplasma hominis and A. laidlawii, PG8 and oral strains, developed typical 'fried-egg' colonies which were remarkably heterogeneous in size. The colonies of A. laidlawii strain NCTC 10116 were more homogeneous and grew mainly on the surface of the agar showing a fine granular appearance. Ureaplasma urealyticum produced smaller, granular colonies which grew deeply embedded in the agar and generally without much surface growth. The cellular ultrastructure in these colonies was also examined. The results indicate that several aspects of colony morphogenesis and ultrastructure varied for each of the three species examined.

Cell fractions and enzymatic activities of Ureaplasma urealyticum.

The localization of some enzymic activities in cell fractions of Ureaplasma urealyticum was studied. A quantitative evaluation of the effectiveness of several cell lysis procedures was obtained by using labeled membranes and sucrose density gradient centrifugation. Ultrasonic treatment was found to be the most effective procedure for lysing the cells, whereas digitonin and osmotic shock caused the lysis of only 70 and 50% of the cells, respectively. The localization of selected enzymes in Ureaplasma cells resembled that found in other Mycoplasma species. Adenosine triphosphatase, ribonuclease, deoxyribonuclease, and p-nitrophenylphosphatase activities were located exclusively in the membrane fraction, whereas urease and L-histidine ammonia-lyase were located in the cytoplasm.

Significance of appropriate techniques and media for isolation and identification of Ureaplasma urealyticum from clinical specimens.

Controversy over the association of Ureaplasma urealyticum with reproductive failure may be due to methods used to isolate the microorganism. U. urealyticum isolations from clinical material should be done simultaneously in broth and on Shepard's differential agar medium (A7) containing manganese sulfate. Urine sediments result in a 9% (P = 0.0002) higher rate of isolation than than cervical and urethral swabs. Primary isolations may not display standard textbook morphology. Isolated colonies may be present, but brown streaks in cervical mucus or a coalescent haze around epithelial cells in urine sediment may also be seen in areas of concentrated growth. The broth and agar media used, method of incubation, type of specimen, and method of storing specimens before culture are all factors which influence the recovery of U. urealyticum.

Morphology and ultrastructure of Ureaplasma urealyticum in agar growth.

Colonies of Ureaplasma urealyticum serotype VI grown on agar were examined by thin sectioning technique. The arrangement of cells within the colonies was less dense and less stratified than that observed in most other mycoplasma colonies. Small, dense and pleomorphic cells were located mainly along the colony borderline towards the agar. Studies of cell morphology after 3, 10 and 15 days of growth revealed that such cells were younger than the less dense, mostly ovoid cells seen in the central area of colonies. The ultrastructure of individual cells was similar to that of Ureaplasma grown in liquid medium, as described earlier, featuring geometrical ribosome arrangements and extramembranous coat. Serial sections demonstrated the difficulties involved in discussing cell morphology on the basis of single thin sections. The advantages of the preparation method used for electron microscopy are discussed.

Multiple forms of urease in cytoplasmic fractions of Ureaplasma urealyticum.

The urease activity of Ureaplasma urealyticum was found to be located in the cytoplasmic fraction and consisted of multiple stable enzyme forms representing at least four biotypes.