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Mollicutes

Mollicutes is a class of bacteria distinguished by the absence of a cell wall and its peptidoglycan. The word Mollicutes is derived from Latin mollis 'soft, pliable' and cutis 'skin'. Individuals are very small, typically only 0.2–0.3 μm in size and have a very small genome size. They vary in form, although most have sterols that make the cell membrane somewhat more rigid. Many move about by gliding, but members of the genus Spiroplasma are helical and move by twisting. The best-known genus in the Mollicutes is Mycoplasma, though it has been split in 2018 into several genera. Colonies show the typical "fried-egg" appearance.

Origin and evolution to parasitic life
Analysis of the genomes of Mollicutes ("mycoplasmas") gives solid support for the hypothesis that Mollicutes evolved from Gram-positive bacteria by a process of reductive evolution. By adopting a parasitic mode of life with use of nutrients from their hosts, many Mollicutes reduced their genetic material considerably. On the other hand, they lost the genes for many assimilative processes. Thus, Mycoplasma possibly became the smallest self-replicating organism in nature. Mycoplasmoides genitalium, with 580,000 base pairs, has an especially small genome size. Some phytoplasmas also have a very small genome size. The genera with the smallest genome are considered to be phylogenetically the most "recent" mollicutes. To maintain their parasitic mode of life the Mollicutes have developed rather sophisticated mechanisms to colonize their hosts and resist the host immune system. Proliferation Mollicutes (formerly also called pleuropneumonia-like organisms, PPLO) as well as L-form bacteria (formerly also called L-phase bacteria), both lacking cell walls, do not proliferate by binary fission but by a budding mechanism. In 1954, this mode of proliferation has been shown by continual observations of live cells using phase-contrast microscopy. Previously, Mollicutes were sometimes considered stable L-form bacteria, but phylogenetic analysis has identified them as bacteria that have lost their cell walls in the course of evolution. ==History of the classification==
History of the classification
The classification of the Mollicutes has always been difficult. The individuals are tiny, and being parasites, they have to be cultivated on special media. Until now, many species could not be isolated at all. In the beginning, whether they were fungi, viruses, or bacteria was not clear. Also, the resemblance to L-forms was confusing. At first, all members of the class Mollicutes were generally named "mycoplasma" or pleuropneumonia-like organism (PPLO). Mollicutes other than some members of genus Mycoplasma were still unidentified. The first species of Mycoplasma/Mollicutes, that could be isolated was Mycoplasma mycoides. This bacterium was cultivated by Nocard and Roux in 1898. In 1956, D.G. Edward and E.A. Freundt made a first proposal for classifying and naming PPLOs. They left undecided, however, whether they belong to the bacteria (prokaryotes, in 1956 called "Schizomycetes") or to the eukaryotes. As type species (name-giving species) of the PPLOs/mycoplasmas, Edward and Freundt proposed Mycoplasma mycoides, being the causative organism of bovine pleuropneumonia and referring to the pleuropneumonia-like organisms. Until then, Mycoplasma mycoides was known as Asterococcus mycoides, but later that name was not recognized as valid. In their publication of 1956, they described 15 species of Mycoplasma. In 1967 the class Mollicutes, containing the order Mycoplasmatales, was proposed by the Subcommittee on Taxonomy of the Mycoplasmata. Now, the name Mycoplasma should exclusively be used for members of the genus Mycoplasma, rather than the use as a trivial name for any Mollicute. As the trivial name has been used in literature for a long time, this is yet not always the case. For classification and nomenclature of Mollicutes, there are special rules, which are maintained by the International Committee on Systematics of Prokaryotes (ICSP) Subcommittee on the Taxonomy of Mollicutes (formerly the International Committee on Systematic Bacteriology (ICSB) Subcommittee on taxonomy of Mycoplasmatales). Traditionally, Mollicutes taxonomy has been based on serology and phenotypic characteristics. However, most modern classifications are based on DNA or RNA sequences, especially 16S rRNA sequences) Bacteria into three divisions (= phyla) on the basis of the cell wall types: • Gram-negative Gracilicutes, with a thin cell wall and little peptidoglycan; • Gram-positive Firmicutes, with a thicker cell wall and more peptidoglycan (the name was later changed in "Firmicutes"), and • the "Mollicutes", without a cell wall. The phylum for Mollicutes The results of Mollicutes phylogenetic analyses have been controversial. Some taxonomists place them in Bacillota, others in Mycoplasmatota. Woese et al. suggested that the Mollicutes might have been derived from different branches of bacteria. They concluded, that the Mollicutes are not a phylogenetically coherent group and therefore do not form a distinct higher level taxon. Instead, they cluster within Gram-positive bacteria of the phylum Bacillota. The results of molecular phylogenetic analyses have been partly dependent on the chosen molecular marker, like rRNA, elongation factor or another protein. Phylogenetic trees based on phosphoglycerate kinase (Pgk) amino acid sequences' indicated a monophyletic origin for the Mollicutes within the Bacillota. An early edition of ''Bergey's Manual of Systematic Bacteriology'' placed class Mollicutes within phylum Bacillota, whereas in the announced 2nd edition, they are moved to a separate phylum Tenericutes (later renamed Mycoplasmatota). The change is motivated by "their unique phenotypic properties, in particular the lack of rigid cell walls, and the general low support by alternative markers". Up-to-date analysis of the SSU (16S) rRNA puts Mollicutes in Bacillota (see LTP below). GTDB's 120-protein approach does the same (see below). The 2018 Mycoplasma split It has been known since the 1990s that Mycoplasma includes several groups that were not very related to its type species. Four lineages were defined by 2007. In 2018, Gupta et al. re-circumscribed the genus Mycoplasma around M. mycoides. A total of 78 species were removed from Mycoplasma, creating five new genera and a number of higher taxonomic levels. Under this new scheme, a new family Mycoplasmoidaceae was created to correspond to the "pneumoniae" group, with M. pneumoniae and related species transferred to a new genus Mycoplasmoides. Another new family Metamycoplasmataceae was created to correspond to the "hominis" group. Both families belong to a new order Mycoplasmoitales, distinct from the Mycoplasmatales of Mycoplasma. The taxonomy was accepted by the ICSB with validation list 184 in 2018 and became the correct name. Both List of Prokaryotic names with Standing in Nomenclature (LPSN) and National Center for Biotechnology Information (NCBI) now use the new nomenclature. Gupta's proposed taxonomy, as expected, moved the medically important "pneumoniae" group out of Mycoplasma into its own genus. As a result, a number of mycoplasmologists petitioned to the ICSB to reject the name in 2019. They argue that although Gupta's phylogenetic methods were likely solid, the proposed name changes are too sweeping to be practically adopted, citing some principles of the Code such as "name stability". Gupta and Oren wrote a rebuttal in 2020, further detailing the pre-existing taxonomic problems. In 2022, the ICSP's Judicial Opinion 122 ruled in favor of the name changes proposed by Gupta, meaning they remain valid under the Prokaryotic Code Gupta et al. 2019 performed some uncontroversial sorting of the order Mycoplasmatales. ==Phylogeny==
Phylogeny
The currently accepted taxonomy is based on the List of Prokaryotic names with Standing in Nomenclature (LPSN) and National Center for Biotechnology Information (NCBI). ♦ Mollicutes Mycoplasmatota and Mollicutes are not listed in GTDB, but they are recovered as a monophyletic node (marked in cladogram) in RS226 with a low non-parametric bootstrap support of 32%. In RS220, Haloplasmataceae was not grouped in under the same node as the rest of the Mollicutes. The taxa labels only reflect an approximate match to the current taxonomy. An alternative match has what is labeled "Mollicutes" (or an inner node excluding Haloplasma, as this genus has a peptidoglycan cell wall) be "Mycoplasmatota", the node labelled "(1)" as Izemoplasmatia, and the node labelled as "(2)" as Mollicutes sensu stricto. == Species which infect humans ==
Species which infect humans
Species in Mollicutes, other than those listed below, have been recovered from humans, but are assumed to have been contracted from a non-human host. The following species use humans as the primary host (modified for 2018 taxonomy): • MalmycoplasmaMa. penetransMetamycoplasmaMe. buccaleMe. fauciumMe. hominisMe. oraleMe. salivariumMycoplasmoidesMd. amphoriformeMd. genitaliumMd. pirumMd. pneumoniaeMycoplasmopsisMp. fermentansM. incognitusMp. lipophilumMp. primatumMp. spermatophilum == Pathogenicity ==
Pathogenicity
Several species in Mollicutes can cause disease, including Md. pneumoniae, which is an important cause of atypical pneumonia (also known as "walking pneumonia"), and Md. genitalium, which has been associated with pelvic inflammatory diseases. Mycoplasma infections in humans are associated with skin eruptions in 17% of cases. P1 antigen The P1 antigen is the primary virulence factor of Mycoplasmoides. P1 is a membrane associated protein that allows adhesion to epithelial cells. The P1 receptor is also expressed on erythrocytes which can lead to autoantibody agglutination from mycobacteria infection. Sexually transmitted infections Ureaplasma, Malmycoplasma, and Metamycoplasma species are not part of the normal vaginal flora. Some Mollicutes species are spread through sexual contact. These species have a negative effect on fertility. In addition, infection is associated with increased risk of cervicitis, infertility, preterm birth, and spontaneous abortion. Mycoplasmoides genitalium has developed resistance to some antibiotics. Infant disease Low birth-weight, preterm infants are susceptible to Ureaplasma, Malmycoplasma, and Metamycoplasma infections. Malmycoplasma and Metamycoplasma species are associated with infant respiratory distress syndrome, bronchopulmonary dysplasia, and intraventricular hemorrhage in preterm infants. These species are: • Mp. fermentansMd. genitaliumMesomycoplasma hyorhinis (Ms. hyorhinis hereafter) • Ma. penetrans The majority of these Mollicutes species have shown a strong correlation to malignant transformation in mammalian cells in vitro. Infection and host cell transformation The presence of Mollicutes was first reported in samples of cancer tissue in the 1960s. as well as the normal contact-dependent inhibition cells. A major feature that differentiates Mollicutes from other carcinogenic pathogens is that the Mollicutes do not cause the cellular changes by insertion of their own genetic material into the host cell.) before the irreversible stage, the infected cells should return to normal. Connections to cancer in vivo and future research Epidemiologic, genetic, and molecular studies suggest infection and inflammation initiate certain cancers, including those of the prostate. Md. genitalium and Ms. hyorhinis induce malignant phenotype in benign human prostate cells (BPH-1) that were not tumorigenic after 19 weeks of exposure. Gastric cancer: Strong evidence indicates the infection of Ms. hyorhinis contributes to the development of cancer within the stomach and increases the likelihood of malignant cancer cell development. Lung cancer: Studies on lung cancer have supported the belief that more than a coincidental positive correlation exists between the appearance of Mollicutes strains in patients and the infection with tumorigenesis. Prostate cancer: p37, a protein encoded for by Ms. hyorhinis, has been found to promote the invasiveness of prostate cancer cells. The protein also causes the growth, morphology, and gene expression of the cells to change, causing them to become a more aggressive phenotype. Renal cancer: Patients with renal cell carcinoma (RCC) exhibited a significantly high amount of Mollicutes sp. compared with the healthy control group. This suggests Mollicutes may play a role in the development of RCC. == Laboratory contaminant ==
Laboratory contaminant
Mollicutes species are often found in research laboratories as contaminants in cell culture. Mollicutes cell culture contamination occurs due to contamination from individuals or contaminated cell culture medium ingredients. A 2002 report lists Me. orale, Ms. hyorhinis, Mp. arginini, Mp. fermentans, Me. hominis, and Acholeplasma laidlawii as the most common contaminants. Mollicutes cells are physically small – less than 1  μm, so are difficult to detect with a conventional microscope. Mollicutes may induce cellular changes, including chromosome aberrations, changes in metabolism, and cell growth. Severe Mollicutes infections may destroy a cell line. Detection techniques include DNA probe, enzyme immunoassays, PCR, plating on sensitive agar, and staining with a DNA stain including DAPI or Hoechst. An estimated 11–15% of U.S. laboratory cell cultures are contaminated with Mollicutes. A Corning study showed that half of U.S. scientists did not test for Mollicutes contamination in their cell cultures. The study also stated that, in former Czechoslovakia, 100% of cell cultures that were not routinely tested were contaminated while only 2% of those routinely tested were contaminated (study p. 6). Since the U.S. contamination rate was based on a study of companies that routinely checked for Mollicutes, the actual contamination rate may be higher. European contamination rates are higher and that of other countries are higher still (up to 80% of Japanese cell cultures). About 1% of published Gene Expression Omnibus data may have been compromised. Several antibiotic-containing formulations of antimycoplasmal reagents have been developed over the years. ==See also==
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