Proposal of Ureaplasma parvum sp. nov. and emended description of Ureaplasma urealyticum (Shepard et al. 1974) Robertson et al. 2001.

The phenotypic and genotypic properties of Ureaplasma urealyticum (family Mycoplasmataceae, order Mycoplasmatales, class Mollicutes) are reviewed here. The 14 recognized serovar standard strains found in humans exhibit no serological cross-reactivity with ureaplasmas from other hosts and uniquely express human immuoglobulin A1 protease activity. However, they exhibit many characteristics which place them in two distinct clusters known as the parvo biovar (or biovar 1 or B) and the T960T biovar (or biovar 2 or A).

A physical and genetic map of the Mycoplasma flocculare ATCC 27716 chromosome reveals large genomic inversions when compared with that of Mycoplasma hyopneumoniae strain J(T).

Pulsed-field gel electrophoresis and DNA hybridization data were used to construct a chromosomal map of Mycoplasma flocculare ATCC 27716, a non-pathogenic inhabitant of porcine respiratory tracts. Twenty-one genetic markers were placed on the map. Comparison of the genetic map with that of the closely related Mycoplasma hyopneumoniae strain J(T), the type strain of the causative agent of enzootic pneumonia in pigs, identified three chromosomal inversions that differentiate these genomes.

A physical and genetic map of the Mycoplasma hyopneumoniae strain J genome.

A macrorestriction map of the genome of Mycoplasma hyopneumoniae strain J, the type strain of the causative agent of enzootic pneumonia in pigs, was constructed using pulsed-field gel electrophoresis (PFGE) and DNA hybridization. The size of the genome as determined by PFGE was approximately 1070 kb. Assembly of the M.

Gene amplification (PCR) to detect and differentiate mycoplasmas in porcine mycoplasmal pneumonia.

The causative agent of porcine mycoplasmal pneumonia, Mycoplasma hyopneumoniae, is difficult and time-consuming to isolate. Serological identification using antibodies induced by the disease is confused by cross-reaction with a closely related organism, Myc. flocculare.

Ureaplasma gallorale, an isolate from chickens, is most closely related to the human isolate, U. urealyticum.

Ureaplasma gallorale is a urease-containing mycoplasma (a member of the Mollicutes) which is pathogenic for chickens, from which it was originally isolated. We amplified the 16S rRNA gene of this bacterium and then cloned and sequenced the amplicon. A phylogenetic analysis based on an alignment of the 16S rRNA sequences of U.

Phylogenetic relationships of the porcine mycoplasmas Mycoplasma hyosynoviae and Mycoplasma hyopharyngis.

The phylogenetic positions of the porcine mycoplasmas Mycoplasma hyosynoviae and Mycoplasma hyopharyngis were determined by using PCR-amplified 16S rRNA gene sequences. M. hyosynoviae is a member of the Mycoplasma hominis group, while M.

An unusual rRNA gene organization in Mycoplasma fermentans (incognitus strain).

The macro-restriction map of Mycoplasma fermentans (incognitus strain) was constructed and its rRNA genes were located on the map. It was found that this organism contains two sets of rRNA genes. The 16S and 23S rRNA genes were closely linked as two clusters.

Comparison of 16S rRNA genes within the T960 and parvo biovars of ureaplasmas isolated from humans.

Two-by-two sequence alignment revealed that the levels of homology between 16S rRNA gene sequences of strains belonging to the two biovars of Ureaplasma urealyticum (class Mollicutes) ranged from 98.5 to 98.9%.

Cloning the ribosomal RNA operons of Mycoplasma flocculare and comparison with those of Mycoplasma hyopneumoniae.

In contrast to other mycoplasma species the 16S/23S rRNA and 5S rRNA operons of Mycoplasma flocculare and Mycoplasma hyopneumoniae map at least 150 kb apart (20% of the genome). Both operons from M. flocculare have been cloned and sequenced.

Differentiation of Mycoplasma hyopneumoniae, M flocculare, and M hyorhinis on the basis of amplification of a 16S rRNA gene sequence.

To differentiate Mycoplasma hyopneumoniae, the cause of mycoplasmal pneumonia in pigs, from M flocculare and M hyorhinis, an assay, using the polymerase chain reaction to amplify a segment of the 16S rRNA gene sequence, was developed. The assay was found to be useful for identification of field isolates, as well as for identification of laboratory-adapted strains. Amplification of DNA from M hyopneumoniae and M flocculare resulted in products of 200 and 400 base pairs, respectively.

Polymerase chain reaction using 16S rRNA gene sequences distinguishes the two biovars of Ureaplasma urealyticum.

Several fundamental phenotypic and genotypic differences have separated strains of the genital mycoplasma Ureaplasma urealyticum into two clusters or biovars. However, the lack of an easily performed and unambiguous test to discriminate between them has hampered investigation of the relationship between these biovars and disease. We determined the 16S rRNA nucleotide sequence of U.

Construction of the physical maps of Mycoplasma hyopneumoniae and Mycoplasma flocculare and the location of rRNA genes on these maps.

The genomes of Mycoplasma flocculare and Mycoplasma hyopneumoniae, two mycoplasmas of the porcine respiratory system, were studied. Based upon antigenic cross-reactivity and DNA-DNA hybridization, these species have given indication of a close genetic relationship. By using field-inversion gel electrophoresis and employing the restriction digest fragments obtained from the gels as the probes, physical maps of the genomes of the two species were constructed.

Phylogenetic relationships of three porcine mycoplasmas, Mycoplasma hyopneumoniae, Mycoplasma flocculare, and Mycoplasma hyorhinis, and complete 16S rRNA sequence of M. flocculare.

The nucleotide sequence of the 16S rRNA gene of Mycoplasma flocculare was determined and was compared with the sequence of a related porcine mycoplasma, Mycoplasma hyopneumoniae. While the overall level of DNA-DNA homology was approximately 11%, sequence alignment of the two 16S rRNA genes yielded a homology value of more than 95%, emphasizing the highly conserved nature of the 16S rRNA gene. Multiple sequence alignments with other mollicutes indicated that M.

The growth response of Mycoplasma hyopneumoniae and Mycoplasma flocculare based upon ATP-dependent luminometry.

Cultures of Mycoplasma hyopneumoniae and M. flocculare in Friis' broth grew faster and to higher titers in air than in 8% CO2; cultures in air grew better when shaken than when stationary. Under the optimal conditions, both species have generation times of about 10 h and achieve maximum titers of at least 10(9) organisms per ml.

Human ureaplasmas show diverse genome sizes by pulsed-field electrophoresis.

Contour clamped homogeneous field (CHEF) agarose gel electrophoresis (AGE), ramped to give linear separation of DNA molecules of 600-1600 kilobase pairs (kbp), was used to determine mobilities for full-sized genomic DNA of the serotype standard strains of the human genital mollicutes, Ureaplasma urealyticum relative to yeast chromosomal DNA markers. Indicated genome sizes (in kbp) were 760 for the four biotype 1 strains and 840-1140 for eleven biotype 2 strains. Other estimates were: 720 for Mycoplasma hominis, 1070 for Mycoplasma hyopneumoniae, 890 for Mycoplasma flocculare, 1180 and 1350 for Mycoplasma mycoides subsp.

A gene probe to detect Mycoplasma hyopneumoniae, the etiological agent of enzootic porcine pneumonia.

Enzootic porcine pneumonia is caused by Mycoplasma hyopneumoniae. Since the disease is of world-wide importance it is important to detect and identify the causative agent. In experience laboratories this mycoplasma can usually be detected by culture but its identification still is difficult and time consuming.

Characterization of tetracycline-resistant strains of Ureaplasma urealyticum.

Gel electrophoresis revealed no plasmids in cloned strains of Ureaplasma urealyticum for which minimal inhibitory concentration (MICs) of tetracycline were greater than 64 mg/l. However, DNA from these strains hybridized with the tetM sequence from Streptococcus agalactiae in dot blot hybridization, whilst DNA from more-susceptible strains did not do so. Our results confirm and extend previous work, in which the tetM sequence was associated with resistance to the tetracycline antibiotics in strains of U.

Septic arthritis due to dual infection with Mycoplasma hominis and Ureaplasma urealyticum.

Acute septic arthritis of a knee and shoulder developed in a 32-year-old renal transplant patient. Cultures yielded Mycoplasma hominis and at least 1, and possibly 2, strains of Ureaplasma urealyticum. Doxycycline therapy controlled the symptoms and signs, and the joints became culture negative.

Purification of urease from Ureaplasma urealyticum.

We have purified urease from the Mollicutes, Ureaplasma urealyticum, using high performance liquid chromatography methods and DEAE-Sephadex chromatography. While only small amounts of material could be utilized in these methods, urease was purified at least 180-fold, yield a major band on SDS-PAGE of 66,000 daltons, a minor band of 64,000 daltons, and several faint bands of lower molecular mass. These results suggest that the 380,000 dalton intact urease is a pentamer or hexamer of these two larger subunits.

ATP measurements obtained by luminometry provide rapid estimation of Ureaplasma urealyticum growth.

ATP content obtained by luciferin-luciferase luminometry with commercially available reagents provided rapid estimates of Ureaplasma urealyticum populations. Each cell contained about 4.7 X 10(-18) mol of ATP.

Effects of anti-Ureaplasma urease antibody on homologous and heterologous urease activities.

Among organisms in the class Mollicutes only Ureaplasma species possess urease. Antiserum to urease of U. urealyticum strain T960 (CX8) was used to examine the cross-reactivity of urease from other Ureaplasma species, as well as urease of jack bean and several urease-possessing walled bacteria.

Serotypes of Ureaplasma urealyticum in spontaneous abortion.

Ureaplasma urealyticum was isolated from the tissues of 63 of 264 (24%) spontaneous abortions obtained by unaided vaginal delivery or by suction/sharp curettage and from 22 of 259 (8%) therapeutic abortions obtained by suction curettage. The isolates were serotyped by a modified indirect colony epifluorescence test. No differences in the incidence of specificities in the test and control groups were apparent, although statistical analysis of the data remains to be performed.

Properties of urease from Ureaplasma urealyticum: kinetics, molecular weight, and demonstration of multiple enzyme isoelectric point forms.

In the several strains of Ureaplasma urealyticum that we examined, all originally isolated from human sources, the ureases were found to have a pH optimum between 7.2 and 7.5, and the Km was approximately 2.

Problems associated with serotyping strains of Ureaplasma urealyticum.

We sought to identify problems associated with serotyping Ureaplasma urealyticum, a human genital tract mycoplasma. We examined the results of serotyping isolates from cases of nongonococcal urethritis and asymptomatic controls and found no indication of correlation between serotype(s) and pathogenic potential. Reproducibility in serotype determination was generally good, i.