-
Microbial Pathogenesis Aug 1995The infectious pattern of mycoplasmas (Mycoplasma penetrans, Mycoplasma pneumoniae and Mycoplasma genitalium) in mammalian cells was examined using confocal microscopy...
The infectious pattern of mycoplasmas (Mycoplasma penetrans, Mycoplasma pneumoniae and Mycoplasma genitalium) in mammalian cells was examined using confocal microscopy and flow cytometry combined with cell fractionation and mycoplasma viability determinations. Within 2 h postinfection mycoplasmas parasitize cell surfaces, enter the intracellular spaces and locate throughout the cytoplasmic and perinuclear regions. These mycoplasmas can be cultivated from cytoplasmic and nuclear fractions 96 h later and continue to persist intracellularly for at least 7 days, suggesting a much more active intracellular role for mycoplasmas than had been considered previously.
Topics: Animals; Cell Fractionation; Cell Line; Flow Cytometry; Humans; Lung; Microscopy, Confocal; Mycoplasma penetrans; Mycoplasma pneumoniae
PubMed: 8577234
DOI: 10.1006/mpat.1995.0050 -
Current Microbiology Jan 2021Mycoplasma is the smallest self-replicating bacteria, figuring as common contaminant of eukaryotic cell cultures. Production inputs and operator's manipulation seem to...
Mycoplasma is the smallest self-replicating bacteria, figuring as common contaminant of eukaryotic cell cultures. Production inputs and operator's manipulation seem to be the main sources of such contamination. Many analytical approaches have been applied for mycoplasma detection in cell cultures and also in biological products. However, unless they were validated, only indicator cell culture and bacteriological culture are considered as compendial methods for quality control of biological products. Nano-flow cytometry has been pointed out as an alternative technique for addressing prokaryotic and eukaryotic cell viability being a substantial tool for reference material production. In this study, a viability-flow-cytometry assay was standardized for M. gallisepticum and then applied to other cell-culture-contaminant mycoplasmas. For this, M. galliseticum's growth rate was observed and different treatments were evaluated to establish low viability cultures (cell death-induced control). Distinct viability markers and their ideal concentrations (titration) were appraised. Ethanol treatment showed to be the best death-inducing control. CFDA and TOPRO markers revealed to be the best choice for detecting live and dead mycoplasma frequencies, respectively. The standardized methodology was applied to Mycoplasma arginini, M. hyorhinis, M. orale, Spiroplasma citri and Acholeplasma laidlawii. Significant statistical difference was observed in the percentage of viable cells in comparison to ethanol treatment for A. laidlawii in CFDA and in both markers for M. gallisepticum, M. hyorhinis and S. citri. In summary, we standardized a flow cytometry assay for assessing M. gallisepticum - and potentially other species - viability and ultimately applied for reference material production improving the quality control of biological products.
Topics: Cell Culture Techniques; Flow Cytometry; Mycoplasma; Mycoplasma gallisepticum; Tenericutes
PubMed: 33159562
DOI: 10.1007/s00284-020-02255-1 -
Scientific Reports Mar 2020Mycoplasma mobile, a fish pathogen, exhibits its own specialized gliding motility on host cells based on ATP hydrolysis. The special protein machinery enabling this...
Mycoplasma mobile, a fish pathogen, exhibits its own specialized gliding motility on host cells based on ATP hydrolysis. The special protein machinery enabling this motility is composed of surface and internal protein complexes. Four proteins, MMOBs 1630, 1660, 1670, and 4860 constitute the internal complex, including paralogs of F-type ATPase/synthase α and β subunits. In the present study, the cellular localisation for the candidate gliding machinery proteins, MMOBs 1620, 1640, 1650, and 5430 was investigated by using a total internal reflection fluorescence microscopy system after tagging these proteins with the enhanced yellow fluorescent protein (EYFP). The M. mobile strain expressing a fusion protein MMOB1620-EYFP exhibited reduced cell-binding activity and a strain expressing MMOB1640 fused with EYFP exhibited increased gliding speed, showing the involvement of these proteins in the gliding mechanism. Based on the genomic sequences, we analysed the sequence conservativity in the proteins of the internal and the surface complexes from four gliding mycoplasma species. The proteins in the internal complex were more conserved compared to the surface complex, suggesting that the surface complex undergoes modifications depending on the host. The analyses suggested that the internal gliding complex was highly conserved probably due to its role in the motility mechanism.
Topics: Bacterial Proteins; Genome, Bacterial; Mycoplasma; Sequence Analysis, DNA
PubMed: 32123220
DOI: 10.1038/s41598-020-60535-z -
Journal of Virological Methods Jan 2022Mollicutes (Mycoplasma and Acholeplasma) are parasitic bacteria that adhere to cellular surfaces, naturally resistant to many antibiotics and extremely small. They are...
Mollicutes (Mycoplasma and Acholeplasma) are parasitic bacteria that adhere to cellular surfaces, naturally resistant to many antibiotics and extremely small. They are often found as contaminants in cultured cells, where they go unnoticed. They may be present in viral stocks because they are present in supernatants of cells where cultured viruses are released. The best way to keep laboratories free of Mycoplasma is to discard infected cultures, but, as judged by the very common finding of Mycoplasma-contaminated cultures in many laboratories, this is not done as often as it should be. A possible reason is that most procedures recommended take as long as performing a simple experiment and many laboratories delay testing to save money and time. Indeed, many methods exist to detect Mycoplasma infection of cell lines, but they take at least a couple of hours of hands-on work, if not more. Here we describe a procedure to screen viral stocks and tissue cultures for Mycoplasma presence. It relies on isolation of Mycoplasma on ordinary horse blood agar directly from exhausted tissue culture supernatants and does not require experienced personnel or expensive equipment. It only requires minutes of hands-on work, and, for this, it may be useful for weekly screening of cultures. It yields semiquantitative results in roughly 5 days, which is the time that usually passes between one subculture passage of cells in vitro to another. Because of its simplicity, it may be useful for detecting Mycoplasma in viral stocks and for frequent screening of cultures in research laboratories.
Topics: Cell Culture Techniques; Cells, Cultured; Humans; Mycoplasma; Mycoplasma Infections; Mycoplasma hyorhinis
PubMed: 34644588
DOI: 10.1016/j.jviromet.2021.114327 -
Research in Veterinary Science Oct 2013Mycoplasma membrane proteins influence cell shape, cell division, motility and adhesion to host cells, and are thought to be integrally involved in the pathogenesis of...
Mycoplasma membrane proteins influence cell shape, cell division, motility and adhesion to host cells, and are thought to be integrally involved in the pathogenesis of mycoplasmoses. Many of the membrane proteins predicted from mycoplasma genome sequences remain hypothetical, as their presence in cellular protein preparations is yet to be established experimentally. Recent genome sequences of several strains of Mycoplasma bovis have provided further insight into the potential role of the membrane proteins of this pathogen in colonisation and infection. This review highlights recent advances in knowledge about the influence of M. bovis membrane proteins on the pathogenesis of infection with this species and identifies future research directions for enhancing our understanding of the role of these proteins.
Topics: Bacterial Outer Membrane Proteins; Gene Expression Regulation, Bacterial; Genome, Bacterial; Mycoplasma bovis; Virulence
PubMed: 23810376
DOI: 10.1016/j.rvsc.2013.05.016 -
Zentralblatt Fur Bakteriologie,... Feb 1973
Topics: Cell Membrane; Culture Media; Mycoplasma; Pseudomonas aeruginosa; Staphylococcus
PubMed: 4145875
DOI: No ID Found -
Veterinaria Italiana Dec 2022This study aimed to perform molecular typing of Mycoplasma mycoides subsp. mycoides from slaughtered cattle in Adamawa and Taraba States, north‑eastern Nigeria. A...
This study aimed to perform molecular typing of Mycoplasma mycoides subsp. mycoides from slaughtered cattle in Adamawa and Taraba States, north‑eastern Nigeria. A total of four hundred and eighty (480) samples of lung tissues, nasal swabs, ear swabs and pleural fluids were collected from cattle at slaughter and processed according to standard laboratory protocols. Identification and confirmation were achieved with specific PCR and PCR‑RFLP. An overall M. mycoides subsp. mycoides isolation rate of 6.87% (33/480) was obtained. In Adamawa State, 12 (10.91%) isolates of M. mycoides subsp. mycoides came from both, lung tissues and pleural fluids. While in Taraba State, 5 (7.14%) and 4 (5.71%) isolates of M. mycoides subsp. mycoides came from lung tissues and pleural fluids, respectively. The samples from nasal and ear swabs from the study states were negative for M. mycoides subsp. mycoides. Thirty‑three out of the 37 culture positive isolates were confirmed to be Mycoplasma mycoides subspecies mycoides with the production of a band equivalent to 574‑bp. Molecular typing with restriction endonuclease Vsp1 results in the two bands of 180‑bp and 380‑bp. In conclusion, the study has established an isolation rate of 6.87% for M. mycoides subsp. mycoides. Measures to strengthen movement control in order to minimise the spread of this dreaded disease of cattle were recommended.
Topics: Animals; Cattle; Mycoplasma mycoides; Nigeria; Mycoplasma; Laboratories
PubMed: 37219834
DOI: 10.12834/VetIt.2331.16051.3 -
G3 (Bethesda, Md.) Feb 2019is an important pathogen involved in bovine respiratory disease, which causes huge economic losses worldwide. Our knowledge regarding the genomics, pathogenic...
is an important pathogen involved in bovine respiratory disease, which causes huge economic losses worldwide. Our knowledge regarding the genomics, pathogenic mechanisms, and genetics of is rather limited. In this study, the complete genome of GS01 strain was sequenced using PacBio SMRT technology and first genome-wide analyzed. GS01 has a single circular chromosome of 1,065,810 bp encoding 825 predicted proteins. Twenty-three potential virulence genes and two pathogenicity islands were identified in This pathogen was cytopathogenic, could form prolific biofilms, and could produce a large amount of HO Methylation analysis revealed adenine and cytosine methylation across the genome and 13 distinct nucleotide motifs. Comparative analysis showed a high collinearity relationship between GS01 and type strain ATCC 27140. Phylogenetic analysis demonstrated that is genetically close to and The data presented in this study will aid further study on the pathogenic mechanisms and evolution of .
Topics: Biofilms; DNA Methylation; Genome, Bacterial; Mycoplasma; Nucleotide Motifs; Phylogeny; Virulence Factors
PubMed: 30573467
DOI: 10.1534/g3.118.200941 -
Klinische Wochenschrift Jul 1961
Topics: Mycoplasma; Mycoplasmataceae; Virulence
PubMed: 13756808
DOI: 10.1007/BF01477786 -
Brazilian Journal of Medical and... 2020The objective of this study was to analyze the infection rate and drug resistance of Ureaplasma urealyticum (UU) and Mycoplasma hominis (MH) in the genitourinary tract...
The objective of this study was to analyze the infection rate and drug resistance of Ureaplasma urealyticum (UU) and Mycoplasma hominis (MH) in the genitourinary tract of Chinese patients. From December 2018 to June 2019, vaginal secretion or urinary secretion of outpatients in our hospital were selected for culture and drug sensitivity analysis of Ureaplasma urealyticum and Mycoplasma hominis. In 4082 Chinese samples, 1567 Mycoplasma were detected, a detection rate of 38.39%, among which 1366 cases were UU single positive, accounting for 33.47%, 15 cases were MH single positive, accounting for 0.36%, 186 cases were UU and MH mixed positive, accounting for 4.56%. The most affected age groups were 21-30 years and 31-40 years, accounting for 19.09 and 15.05%, respectively. The results of drug sensitivity showed that doxycycline, minocycline, josamycin, clarithromycin, and roxithromycin were more sensitive to mycoplasma infection. The distribution of Ureaplasma urealyticum and Mycoplasma hominis in the human genitourinary system and their sensitivity to antibiotics is different for sex and age groups.
Topics: Adult; Anti-Bacterial Agents; Asian People; China; Female; Humans; Male; Microbial Sensitivity Tests; Mycoplasma hominis; Ureaplasma Infections; Ureaplasma urealyticum; Young Adult
PubMed: 33263642
DOI: 10.1590/1414-431X202010099