A phylogenomic and molecular markers based taxonomic framework for members of the order Entomoplasmatales: proposal for an emended order Mycoplasmatales containing the family Spiroplasmataceae and emended family Mycoplasmataceae comprised of six genera.

The "Spiroplasma cluster" is a taxonomically heterogeneous assemblage within the phylum Tenericutes encompassing different Entomoplasmatales species as well as the genus Mycoplasma, type genus of the order Mycoplasmatales. Within this cluster, the family Entomoplasmataceae contains two non-cohesive genera Entomoplasma and Mesoplasma with their members exhibiting extensive polyphyletic branching; additionally, the genus Mycoplasma is also embedded within this family. Genome sequences are now available for all 19 Entomoplasmataceae species with validly published names, as well as 6 of the 7 species from the genus Mycoplasma. With the aim of developing a reliable phylogenetic and taxonomic framework for the family Entomoplasmataceae, exhaustive phylogenetic and comparative genomic studies were carried out on these genome sequences. Phylogenetic trees were constructed based on concatenated sequences of 121 core proteins for this cluster, 67 conserved proteins shared with the phylum Firmicutes, 40 ribosomal proteins, three major subunits of RNA polymerase (RpoA, B and C) by different means and also for the 16S rRNA gene sequences. The interspecies relationships as well as different species groups observed in these trees were identical and robustly resolved. In all of these trees, members of the genera Mesoplasma and Entomoplasma formed three and two distinct clades, respectively, which were interspersed among the members of the other genus. The observed species groupings in the phylogenetic trees are independently strongly supported by our identification of 103 novel molecular markers or synapomorphies in the forms of conserved signature indels and conserved signature proteins, which are uniquely shared by the members of different observed species clades. To account for the different observed species clades, we are proposing a division of the genus Mesoplasma into an emended genus Mesoplasma and two new genera Tullyiplasma gen. nov. and Edwardiiplasma gen. nov. Likewise, to recognize the distinct species groupings of Entomoplasma, we are proposing its division into an emended genus Entomoplasma and a new genus Williamsoniiplasma gen. nov. Lastly, to rectify the long-existing taxonomic anomaly caused by the presence of genus Mycoplasma (order Mycoplasmatales) within the Entomoplasmatales, we are proposing an emendation of the family Mycoplasmataceae to include both Entomoplasmataceae plus Mycoplasma species and an emendation of the order Mycoplasmatales, which now comprises of the emended family Mycoplasmataceae and the family Spiroplasmataceae. The taxonomic reclassifications proposed here accurately reflect the species relationships within this group of Tenericutes and they should lead to a better understanding of their biological and pathogenic characteristics.

Mycoplasmateceae species are not found in fallopian tubes of women with tubo-peritoneal infertility

BACKGROUND: The role of mycoplasmas on the development and sequelae of pelvic inflammatory disease remains controversial. The objective of the present study is to correlate directly the presence of Mycoplasmateceae through polimerase chain reaction (PCR) determinations in cervix and Fallopian tubes of infertile patients with tubo-peritoneal factor diagnosed through laparoscopy. METHODS: Thirty patients with tubo-peritoneal infertility and 30 normal fertile patients were included in the study; cervical samples and tubal flushings were obtained during laparoscopy. PCR determinations for the detection of genetic material of Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealiticum, Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis in cervix and tubal flushings were performed. RESULTS: No Mycoplasmataceae species as "only" microorganisms were found in tubal flushings of tubo-peritoneal infertility patients, whereas three (10%) fertile patients with normal tubes were positive for mycoplasma presence. This difference was not significant (p = 0.237). Among the 30 patients suffering from tubal infertility diagnosed through laparoscopy, Mycoplasmatecae species were not detected in the Fallopian tubes by PCR determinations, while in normal tubes from fertile patients these and other microorganisms could be found without distorting tubal anatomy. CONCLUSION: Mycoplasmateceae species were not detected in Fallopian tubes of women with tubo-peritoneal infertility.

[Evaluation of the Mycoscreen technique for the detection and quantification of genital mycoplasmas]. / Evaluación del método Mycoscreen para la detección y cuantificación de micoplasmas genitales.

A total of 161 samples from genital sources were evaluated using two different methods for genital Mycoplasma isolation: inoculation to urea broth, arginine broth and A7 solid media (standard method) and a new enzymatic method (Mycoscreen Oxoid). The isolation rates for Ureaplasma urealitycum were 42% (standard method) and 49% (Mycoscreen) (sensitivity: 100%; specificity: 88%). The isolation rates for Mycoplasma hominis were 13% (standard method) and 14% (Mycoscreen) (sensitivity: 80%; specificity: 95%). Using the enzymatic method, manufacturer's positive result is defined as a count greater than 10,000 UCC-ml at 24 hours. When we compare the 24 hours results, the concordance rates was 84% (Ureaplasma urealyticum) and 62% (Mycoplasma hominis). Mycoscreen method is a valid and fast technique for the isolation and identification of genital Mycoplasma. It also allows a fast screening of pathologic counts of Ureaplasma urealyticum in genital exudates.

Urine as a specimen for diagnosis of sexually transmitted diseases.

The case of specimen collection has led to the suggestion that urine might be a useful specimen for the isolation of sexually transmitted disease agents. It would only be an appropriate specimen for agents that infect the urethra, such as Neisseria gonorrhoeae or Chlamydia trachomatis. Comparative tests have shown that culture of urine for chlamydiae (from men with urethritis) or for gonococci from women is an insensitive procedure. Gonococci can be isolated from urine from men at rates essentially equivalent to culture of urethral swabs. If specimens can be processed promptly (to avoid bactericidal effects of urine), culture of urine can likely be useful for screening asymptomatic men for gonococcal infection.