Rheotaxis in Mycoplasma gliding.

This review describes the upstream-directed movement in the small parasitic bacterium Mycoplasma. Many Mycoplasma species exhibit gliding motility, a form of biological motion over surfaces without the aid of general surface appendages such as flagella. The gliding motility is characterized by a constant unidirectional movement without changes in direction or backward motion. Unlike flagellated bacteria, Mycoplasma lacks the general chemotactic signaling system to control their moving direction. Therefore, the physiological role of directionless travel in Mycoplasma gliding remains unclear. Recently, high-precision measurements under an optical microscope have revealed that three species of Mycoplasma exhibited rheotaxis, that is, the direction of gliding motility is lead upstream by the water flow. This intriguing response appears to be optimized for the flow patterns encountered at host surfaces. This review provides a comprehensive overview of the morphology, behavior, and habitat of Mycoplasma gliding, and discusses the possibility that the rheotaxis is ubiquitous among them.

Seroprevalence of mycoplasmosis in broiler, layer, and native chickens in Giza, Egypt.

We aimed to investigate Mycoplasma infections among chicken flocks (Ross, Lohmann and native) in Giza, Egypt, using serological tests, including the slide plate agglutination (SPA) test, hemagglutination inhibition (HI) test, and enzyme-linked immunosorbent assay (ELISA). The slide plate agglutination examination, a serological test, indicated the prevalence of Mg and Ms infections of 10.9% and 13.2%, respectively. On 91 SPA test positive serum samples for either Mg or Ms, a passive hemagglutination/hemagglutination inhibition (HI) test was performed. The SPA and HI test findings were found to be comparable. On 90 SPA test positive samples, an ELISA was performed using commercial kits for Mg and Ms serodiagnosis. According to the ELISA data, only 83.33% and 18.88% of SPA test positive samples were confirmed as positive for Ms and Mg infections, respectively. The prevalence increased to 84.44% and 77.77%, respectively, when suspected samples were deemed positive.

Movements of Mycoplasma mobile Gliding Machinery Detected by High-Speed Atomic Force Microscopy.

Mycoplasma mobile, a parasitic bacterium, glides on solid surfaces, such as animal cells and glass, by a special mechanism. This process is driven by the force generated through ATP hydrolysis on an internal structure. However, the spatial and temporal behaviors of the internal structures in living cells are unclear. In this study, we detected the movements of the internal structure by scanning cells immobilized on a glass substrate using high-speed atomic force microscopy (HS-AFM). By scanning the surface of a cell, we succeeded in visualizing particles, 2 nm in height and aligned mostly along the cell axis with a pitch of 31.5 nm, consistent with previously reported features based on electron microscopy. Movements of individual particles were then analyzed by HS-AFM. In the presence of sodium azide, the average speed of particle movements was reduced, suggesting that movement is linked to ATP hydrolysis. Partial inhibition of the reaction by sodium azide enabled us to analyze particle behavior in detail, showing that the particles move 9 nm right, relative to the gliding direction, and 2 nm into the cell interior in 330 ms and then return to their original position, based on ATP hydrolysis. IMPORTANCE The Mycoplasma genus contains bacteria generally parasitic to animals and plants. Some Mycoplasma species form a protrusion at a pole, bind to solid surfaces, and glide by a special mechanism linked to their infection and survival. The special machinery for gliding can be divided into surface and internal structures that have evolved from rotary motors represented by ATP synthases. This study succeeded in visualizing the real-time movements of the internal structure by scanning from the outside of the cell using an innovative high-speed atomic force microscope and then analyzing their behaviors.

Establishment of caprine airway epithelial cells grown in an air-liquid interface system to study caprine respiratory viruses and bacteria.

Respiratory diseases negatively impact the global goat industry, but are understudied. There is a shortage of established and biological relevant in vitro or ex vivo assays to study caprine respiratory infections. Here, we describe the establishment of an in vitro system based on well-differentiated caprine airway epithelial cell (AEC) cultures grown under air liquid interface conditions as an experimental platform to study caprine respiratory pathogens. The functional differentiation of the AEC cultures was monitored and confirmed by light and immunofluorescence microscopy, scanning electron microscopy and examination of histological sections. We validated the functionality of the platform by studying Influenza D Virus (IDV) infection and Mycoplasma mycoides subsp. capri (Mmc) colonization over 5 days, including monitoring of infectious agents by titration and qPCR as well as colour changing units, respectively. The inoculation of caprine AEC cultures with IDV showed that efficient viral replication takes place, and revealed that IDV has a marked cell tropism for ciliated cells. Furthermore, AEC cultures were successfully infected with Mmc using a multiplicity of infection of 0.1 and colonization was monitored over several days. Altogether, these results demonstrate that our newly-established caprine AEC cultures can be used to investigate host-pathogen interactions of caprine respiratory pathogens.

Mycoplasma agassizii, an opportunistic pathogen of tortoises, shows very little genetic variation across the Mojave and Sonoran Deserts.

Mycoplasma agassizii is a common cause of upper respiratory tract disease in Mojave desert tortoises (Gopherus agassizii). So far, only two strains of this bacterium have been sequenced, and very little is known about its patterns of genetic diversity. Understanding genetic variability of this pathogen is essential to implement conservation programs for their threatened, long-lived hosts. We used next generation sequencing to explore the genomic diversity of 86 cultured samples of M. agassizii collected from mostly healthy Mojave and Sonoran desert tortoises in 2011 and 2012. All samples with enough sequencing coverage exhibited a higher similarity to M. agassizii strain PS6T (collected in Las Vegas Valley, Nevada) than to strain 723 (collected in Sanibel Island, Florida). All eight genomes with a sequencing coverage over 2x were subjected to multiple analyses to detect single-nucleotide polymorphisms (SNPs). Strikingly, even though we detected 1373 SNPs between strains PS6T and 723, we did not detect any SNP between PS6T and our eight samples. Our whole genome analyses reveal that M. agassizii strain PS6T may be present across a wide geographic extent in healthy Mojave and Sonoran desert tortoises.

Contagious caprine pleuropneumonia (CCPP): A systematic review and meta-analysis of the prevalence in sheep and goats.

Contagious caprine pleuropneumonia (CCPP) is a highly contagious respiratory disease of small ruminants that is caused by the bacterium Mycoplasma capricolum subsp. capripneumoniae. Sheep and goats are two of the species of small ruminants most at risk of CCPP. Outbreaks of CCPP cause significant economic and trade disturbances in several parts of the world. However, the extent and magnitude of CCPP in a particular geographical region is still not well known due to lack of comprehensive data on its occurrence. The present study aimed to investigate the prevalence of CCPP in sheep and goats raised in different geographical regions as well as the factors contributing to the spread of the disease. Searches were done in five online repositories: PubMed, Web of Science, Scopus, CAB Direct and Google Scholar using pre-selected key terms. Data were retrieved from the 41 articles that met the study's inclusion criteria. The pooled prevalences were determined using a random effect meta-analysis model. Prevalence of CCPP was 23.19% (95% CI: 11.90-34.47%) in sheep and 24.91% (95% CI: 20.99-28.84%) in goats. Overall, the regional level pooled prevalence estimates ranged from 8.0% (95% CI: 6.91-9.09%) to 28.70% (22.02-35.38%), depending on species and world region. Substantial heterogeneity (I2  > 75%) was observed in most pooled prevalence estimates. The results indicate high prevalences of CCPP in sheep and goats across the regions, particularly in Africa and Asia, and highlights the need to institute appropriate control measures. Active surveillance and research on risk factors are recommended.

'Candidatus Mycoplasma haemoalbiventris', a novel hemoplasma species in white-eared opossums (Didelphis albiventris) from Brazil.

Opossums of the genus Didelphis are considered synanthropic animals due to their close contact with human beings. Previously, two species of hemotropic mycoplasmas (hemoplasmas) have been detected in opossums: 'Candidatus Mycoplasma haemodidelphidis' in the North American opossum (Didelphis virginiana) and a potentially novel hemotropic Mycoplasma sp. in the white-eared opossums (Didelphis albiventris) from Brazil. Accordingly, the aims of this study were as follows: (a) to determine the prevalence of hemotropic Mycoplasma spp. in free-ranging opossums, (b) to characterize molecularly the hemotropic Mycoplasma sp. infecting opossums and (c) to determine factors associated with hemoplasma infection in opossums from Canoinhas municipality, Santa Catarina State, southern Brazil. For this purpose, 50 white-eared opossums (33 captured and 17 road-killed animals) were evaluated by a pan-hemoplasma PCR assay based on 16S rRNA. Six out of 50 (12%; 95% CI: 5.6%-23.8%) opossums were infested by Ctenocephalides felis fleas. Twenty out of 50 (40%; 95% CI: 26.41%-54.82%) opossums tested positive for hemotropic Mycoplasma sp. by PCR. Sequencing and phylogenetic analysis of the 16S and 23S rRNA gene fragments confirmed that animals were infected by a potentially novel hemotropic Mycoplasma sp. previously reported in white-eared opossums from Brazil. No significant association was found between gender (p = .7759), trap area (p = .0887) or presence of fleas (p = .3811) and positivity for hemoplasmas. The potentially novel hemoplasma species seems to be highly prevalent in white-eared opossums from the states of Paraná, Santa Catarina and Mato Grosso do Sul. Based on the phylogenetic analyses of the 16S rRNA and 23S rRNA genes along with epidemiological data, the name 'Candidatus Mycoplasma haemoalbiventris' is proposed for this novel organism.

The Enemy Within: How Does a Bacterium Inhibit the Foraging Aptitude and Risk Management Behavior of Allenby's Gerbils?

AbstractMicrobes inhabiting multicellular organisms have complex, often subtle effects on their hosts. Gerbillus andersoni allenbyi are commonly infected with Mycoplasma haemomuris-like bacteria, which may cause mild nutrient (choline, arginine) deficiencies. However, are there more serious ecological consequences of infection, such as effects on foraging aptitudes and risk management? We tested two alternatives: the nutrient compensation hypothesis (does nutrient deficiency induce infected gerbils to make up for the shortfall by foraging more and taking greater risks?) and (2) the lethargy hypothesis (do sick gerbils forage less, and are they compromised in their ability to detect predators or risky microhabitats?). We compared the foraging and risk management behavior of infected and noninfected gerbils. We experimentally infected gerbils with the bacteria, which allowed us to compare between noninfected, acutely infected (peak infection loads), and chronically infected (low infection loads) individuals. Our findings supported the lethargy hypothesis over the nutrient compensation hypothesis. Infected individuals incurred dramatically elevated foraging costs, including less efficient foraging, diminished "quality" of time spent vigilant, and increased owl predation. Interestingly, gerbils that were chronically infected (lower bacteria load) experienced larger ecological costs than acutely infected individuals (i.e., peak infection loads). This suggests that the debilitating effects of infection occur gradually, with a progressive decline in the quality of time gerbils allocated to foraging and managing risk. These increased long-term costs of infection demonstrate how small direct physiological costs of infection can lead to large indirect ecological costs. The indirect ecological costs of this parasite appear to be much greater than the direct physiological costs.

Reproduction of contagious bovine pleuropneumonia via aerosol-based challenge with Mycoplasma mycoides subsp. mycoides.

Contagious bovine pleuropneumonia (CBPP) is a respiratory disease caused by Mycoplasma mycoides subsp. mycoides. Infection occurs via Mycoplasma-containing droplets and therefore requires close contact between animals. The current infection models are suboptimal and based on intratracheal installation of mycoplasmas or in-contact infection. This work tested the infection of adult cattle via aerosols containing live mycoplasmas mimicking the infection of cattle in the field. Therefore, we infected six cattle with aerosolized Mycoplasma mycoides subsp. mycoides strain Afadé over seven consecutive days with altogether 109 colony forming units. All animals seroconverted between 11-24 days post infection and five out of six animals showed typical CBPP lesions. One animal did not show any lung lesions at necropsy, while another animal had to be euthanized at 25 days post infection because it reached endpoint criteria. Seroconversion confirmed successful infection and the spectrum of clinical and lesions observed mirrors epidemiological models and the field situation, in which only a fraction of animals suffers from acute clinical disease post infection.

The Lipocalin2 Gene is Regulated in Mammary Epithelial Cells by NFκB and C/EBP In Response to Mycoplasma.

Lcn2 gene expression increases in response to cell stress signals, particularly in cells involved in the innate immune response. Human Lcn2 (NGAL) is increased in the blood and tissues in response to many stressors including microbial infection and in response to LPS in myeloid and epithelial cells. Here we extend the microbial activators of Lcn2 to mycoplasma and describe studies in which the mechanism of Lcn2 gene regulation by MALP-2 and mycoplasma infection was investigated in mouse mammary epithelial cells. As for the LPS response of myeloid cells, Lcn2 expression in epithelial cells is preceded by increased TNFα, IL-6 and IκBζ expression and selective reduction of IκBζ reduces Lcn2 promoter activity. Lcn2 promoter activation remains elevated well beyond the period of exposure to MALP-2 and is persistently elevated in mycoplasma infected cells. Activation of either the human or the mouse Lcn2 promoter requires both NFκB and C/EBP for activation. Thus, Lcn2 is strongly and enduringly activated by mycoplasma components that stimulate the innate immune response with the same basic regulatory mechanism for the human and mouse genes.

Bacteria Boost Mammalian Host NAD Metabolism by Engaging the Deamidated Biosynthesis Pathway.

Nicotinamide adenine dinucleotide (NAD), a cofactor for hundreds of metabolic reactions in all cell types, plays an essential role in metabolism, DNA repair, and aging. However, how NAD metabolism is impacted by the environment remains unclear. Here, we report an unexpected trans-kingdom cooperation between bacteria and mammalian cells wherein bacteria contribute to host NAD biosynthesis. Bacteria confer resistance to inhibitors of NAMPT, the rate-limiting enzyme in the amidated NAD salvage pathway, in cancer cells and xenograft tumors. Mechanistically, a microbial nicotinamidase (PncA) that converts nicotinamide to nicotinic acid, a precursor in the alternative deamidated NAD salvage pathway, is necessary and sufficient for this protective effect. Using stable isotope tracing and microbiota-depleted mice, we demonstrate that this bacteria-mediated deamidation contributes substantially to the NAD-boosting effect of oral nicotinamide and nicotinamide riboside supplementation in several tissues. Collectively, our findings reveal an important role of bacteria-enabled deamidated pathway in host NAD metabolism.

Effects of famciclovir in cats with spontaneous acute upper respiratory tract disease.

OBJECTIVES: The aim of this study was to assess the effects of famciclovir administration in cats with spontaneously acquired acute upper respiratory tract disease. METHODS: Twenty-four kittens with clinical signs of acute upper respiratory tract disease were randomly allocated to receive doxycycline (5 mg/kg PO q12h) alone (group D; n = 12) or with famciclovir (90 mg/kg PO q12h; group DF; n = 12) for up to 3 weeks. Clinical disease severity was scored at study entry and daily thereafter. Oculo-oropharyngeal swabs collected at study entry and exit were assessed using quantitative PCR for nucleic acids of feline herpesvirus type 1 (FHV-1), feline calicivirus (FCV), Chlamydia felis, Bordetella bronchiseptica and Mycoplasma felis. RESULTS: The median (range) age of cats was 1.5 (1-6) months in group D vs 1.6 (1-5) months in group DF (P = 0.54). Pathogens detected in oculo-oropharyngeal swabs at study entry included FCV (n = 13/24; 54%), M felis (n = 8/24; 33%), FHV-1 (n = 7/24; 29%), C felis (n = 7/24; 29%) and B bronchiseptica (n = 3/24; 12%). Median (range) duration of clinical signs was 11.5 (3-21) days in group DF and 11 (3-21) days in group D (P = 0.75). Median (range) total disease score at the end of the study did not differ between groups (group D 1 [1-1] vs group DF 1 [1-3]; P = 0.08). CONCLUSIONS AND RELEVANCE: This study revealed no significant difference in response to therapy between cats treated with doxycycline alone or with famciclovir; cats improved rapidly in both groups. However, identification of FHV-1 DNA was relatively uncommon in this study and clinical trials focused on FHV-1-infected cats are warranted to better evaluate famciclovir efficacy.

Phenotypic diversity in an emerging mycoplasmal disease.

The avian pathogen Mycoplasma gallisepticum (MG) is a known pathogen of poultry, and newly emerged pathogen of house finches wherein it is associated with lethal conjunctivitis. Factors present in MG that are known to mediate virulence include cytadherence, sialidase activity, peroxide production, and biofilm formation. We have quantitatively assessed these factors for MG isolates from house finches from a temporal and geographic distribution across the continental United States that show differing capacity for virulence in vivo. Statistically significant (P < 0.05) differences were observed across strains for sialidase activity, cytadherence, and hydrogen peroxide production. Sialidase activity increased over time in geographically static populations, but did not correlate with time overall. All strains were able to bind α-2,6-linked sialic acid. No strains were found to bind α-2,3-linked sialic acid. All strains produced biofilms in vitro; however, no significant differences were observed in the density of biofilms across strains. Quantitative variance in virulence-associated traits is consistent with within-host evolutionary adaptation in response to a change in ecological niche by a parasitic pathogen.

Mycoplasma nasistruthionis sp. nov. and Mycoplasma struthionis sp. nov. isolated from ostriches with respiratory disease.

Twelve Mycoplasma (M.) strains isolated from the nose, the trachea, and the lung of ostriches (Struthio camelus) displaying respiratory disease were investigated. Analysis of 16S rRNA gene sequences placed five of these strains within the M. synoviae cluster, and seven strains within the M. hominis cluster of genus Mycoplasma, which was further confirmed by analyses of the 16S-23S rRNA intergenic spacer region, and partial rpoB gene and amino acid sequences. Genomic information as well as phenotypic features obtained by matrix-assisted laser desorption ionization time of flight (MALDI-ToF) mass spectrometry analysis and serological reactions indicated that the strains examined are representatives of two hitherto unclassified species of genus Mycoplasma, for which the names Mycoplasma nasistruthionis sp. nov., with type strain 2F1AT (= ATCC BAA-1893T = DSM 22456T), and Mycoplasma struthionis sp. nov., with type strain 237IAT (= ATCC BAA-1890T = DSM 22453T), are proposed.

Evaluation of the immunogenicity and efficacy of BCG and MTBVAC vaccines using a natural transmission model of tuberculosis.

Effective vaccines against tuberculosis (TB) are needed in order to prevent TB transmission in human and animal populations. Evaluation of TB vaccines may be facilitated by using reliable animal models that mimic host pathophysiology and natural transmission of the disease as closely as possible. In this study, we evaluated the immunogenicity and efficacy of two attenuated vaccines, BCG and MTBVAC, after each was given to 17 goats (2 months old) and then exposed for 9 months to goats infected with M. caprae. In general, MTBVAC-vaccinated goats showed higher interferon-gamma release than BCG vaccinated goats in response to bovine protein purified derivative and ESAT-6/CFP-10 antigens and the response was significantly higher than that observed in the control group until challenge. All animals showed lesions consistent with TB at the end of the study. Goats that received either vaccine showed significantly lower scores for pulmonary lymph nodes and total lesions than unvaccinated controls. Both MTBVAC and BCG vaccines proved to be immunogenic and effective in reducing severity of TB pathology caused by M. caprae. Our model system of natural TB transmission may be useful for evaluating and optimizing vaccines.

Differential interaction of the dark septate endophyte Cadophora sp. and fungal pathogens in vitro and in planta.

Dark septate endophytes (DSEs) present a group of widespread root-colonizing fungi. The role of these endophytes in ecosystems and their interactions with plant pathogens are not well understood. In the current study, we assessed the antagonistic potential of the model DSE Cadophora sp. against the tomato soilborne pathogens Rhizoctonia solani, Pythium aphanidermatum and Verticillium dahliae. To investigate their interactions, we conducted in vitro assays followed by a greenhouse experiments in which tomato plants were inoculated with different combinations of the DSE and pathogens. RNA accumulation of selected tomato pathogenesis-related genes and of Cadophora sp. genes with putative antifungal function was analyzed. Cadophora sp. inhibited the growth of the fungal pathogens in vitro and vice versa; a negative impact of the pathogens on the growth of the DSE was also detected. In roots, however, this mutual negative interaction could not be observed. Expression analyses of plant genes could not explain this differential effect, but among the Cadophora sp. genes analyzed, a gene coding for a chalcone synthase was downregulated in planta. The data indicate that plants can change the interaction between fungi and, therefore, in vitro detected antagonism does not necessarily reflect the situation inside the plant.

Molecular and biological properties of phytoplasmas.

Phytoplasmas, a large group of plant-pathogenic, phloem-inhabiting bacteria were discovered by Japanese scientists in 1967. They are transmitted from plant to plant by phloem-feeding insect hosts and cause a variety of symptoms and considerable damage in more than 1,000 plant species. In the first quarter century following the discovery of phytoplasmas, their tiny cell size and the difficulty in culturing them hampered their biological classification and restricted research to ecological studies such as detection by electron microscopy and identification of insect vectors. In the 1990s, however, tremendous advances in molecular biology and related technologies encouraged investigation of phytoplasmas at the molecular level. In the last quarter century, molecular biology has revealed important properties of phytoplasmas. This review summarizes the history and current status of phytoplasma research, focusing on their discovery, molecular classification, diagnosis of phytoplasma diseases, reductive evolution of their genomes, characteristic features of their plasmids, molecular mechanisms of insect transmission, virulence factors, and chemotherapy.

The dynamics between limited-term and lifelong coinfecting bacterial parasites in wild rodent hosts.

Interactions between coinfecting parasites may take various forms, either direct or indirect, facilitative or competitive, and may be mediated by either bottom-up or top-down mechanisms. Although each form of interaction leads to different evolutionary and ecological outcomes, it is challenging to tease them apart throughout the infection period. To establish the first step towards a mechanistic understanding of the interactions between coinfecting limited-term bacterial parasites and lifelong bacterial parasites, we studied the coinfection of Bartonella sp. (limited-term) and Mycoplasma sp. (lifelong), which commonly co-occur in wild rodents. We infected Bartonella- and Mycoplasma-free rodents with each species, and simultaneously with both, and quantified the infection dynamics and host responses. Bartonella benefited from the interaction; its infection load decreased more slowly in coinfected rodents than in rodents infected with Bartonella alone. There were no indications for bottom-up effects, but coinfected rodents experienced various changes, depending on the infection stage, in their body mass, stress levels and activity pattern, which may further affect bacterial replication and transmission. Interestingly, the infection dynamics and changes in the average coinfected rodent traits were more similar to the chronic effects of Mycoplasma infection, whereas coinfection uniquely impaired the host's physiological and behavioral stability. These results suggest that parasites with distinct life history strategies may interact, and their interaction may be asymmetric, non-additive, multifaceted and dynamic through time. Because multiple, sometimes contrasting, forms of interactions are simultaneously at play and their relative importance alternates throughout the course of infection, the overall outcome may change under different ecological conditions.

Ameliorative effect of synthesized silver nanoparticles by green route method from Zingiber zerumbet on mycoplasmal pneumonia in experimental mice.

Mycoplasma pneumoniae (MP) can infect both the upper and lower respiratory tracts related diseases and a common cause of community-acquired pneumonia, mostly in young children and adolescents. Zingiber zerumbet (L.) belongs to the family of Zingiberaceae and it is a perennial and aromatic plant that cultivates in subtropical and tropical countries. This plant is traditionally found throughout Southeast Asia and the Pacific region, where it is commonly used in foods, and beverages purposes. Zingiber zerumbet is a valuable foundation of diverse classes of bioactive major compounds that fit to a varied diversity of chemical metabolites, including polyphenols, alkaloids and terpenes. The numerous studies of Z. zerumbet have shown the enormous pharmacological potential of this plant and its derived bioactive compounds in the treatment of various immune-related diseases like inflammation and other chronic diseases. Based on the previous scientific reports, there are no scientific investigations that claim the antipneumonial activity of the Z. zerumbet based silver nanoparticle. Therefore, the aim of the present study was designed and evaluated the anti-pneumonial potential of biosynthesized with Z. zerumbet based silver nanoparticles in mycoplasmal pneumonia in experimental rats.

First description of two moderately halophilic and psychrotolerant Mycoplasma species isolated from cephalopods and proposal of Mycoplasma marinum sp. nov. and Mycoplasma todarodis sp. nov.

Two moderately halophilic and psychrotolerant new Mycoplasma species were isolated from common cephalopods. Three strains were isolated in pure culture from two individual European flying squid (Todarodes sagittatus), and two individual octopuses (Octopus vulgaris). The strains showed optimal growth at 25 °C and a salinity of 3% (w/v) NaCl. Molecular analyses revealed that the isolates belonged to two new, but phylogenetically related species, divergent from all previously described Mollicutes, representing the first marine isolates of the class, and also the first Mycoplasma strains for which NaCl requirement has been demonstrated. A genome search against all available marine metagenomes and 16S rRNA gene databases indicated that these two species represent a novel non-free-living marine lineage of Mollicutes, specifically associated with marine animals. Morphology and physiology were compatible with other members of this group, and genomic and phenotypic analyses demonstrated that these organisms represent two novel species of the genus Mycoplasma, for which the names Mycoplasma marinum sp. nov. and Mycoplasma todarodis sp. nov. are proposed; the type strains are PET (DSM 105487T, CIP 111404T) and 5HT (DSM 105,488T, CIP 111405T), respectively.