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Nihon Saikingaku Zasshi. Japanese... 2015Bacteria have various way to move over solid surfaces, such as glass, agar, and host cell. These movements involve surface appendages including flagella, type IV pili... (Review)
Review
Bacteria have various way to move over solid surfaces, such as glass, agar, and host cell. These movements involve surface appendages including flagella, type IV pili and other "mysterious" nano-machineries. Gliding motility was a term used various surface movements by several mechanisms that have not been well understood in past few decades. However, development of visualization techniques allowed us to make much progress on their dynamics of machineries. It also provided us better understanding how bacteria move over surfaces and why bacteria move in natural environments. In this review, I will introduce recent studies on the gliding motility of Flavobacteium and Mycoplasma based on the detail observation of single cell and its motility machinery with micro-nano scales.
Topics: Bacterial Translocation; Fimbriae, Bacterial; Flagella; Flavobacterium; Mycoplasma
PubMed: 26632217
DOI: 10.3412/jsb.70.375 -
Archives of Razi Institute Mar 2021Mycoplasma agalactiae (M. agalactiae) is known as the main etiological agent of contagious agalactia (CA). The CA is a disease affecting dairy sheep and goats, the main...
Mycoplasma agalactiae (M. agalactiae) is known as the main etiological agent of contagious agalactia (CA). The CA is a disease affecting dairy sheep and goats, the main characteristics of which include keratoconjunctivitis, arthritis, and mastitis. This pathogen results in milk production reduction and suppression, thereby leading to serious economic loss. In the present study, 125 sheep and goat samples were collected from 15 provinces of Iran. Cultural and molecular methods were used for sample characterization. After extracting genomic DNAs using the phenol/chloroform method, the PCR technique was employed to detect Mycoplasma genus in 163bp fragment of 16S rRNA gene (M-PCR) and M. agalactiae in 800bp fragment of conserve and specific P30 lipoprotein gene (P30-PCR) in cultural and clinical samples. Finally, to validate the experimental approach, a 375 bp amplicon of P80 lipoprotein was amplified using the MA-PCR. Out of 125 samples under investigation, 43 cases were positive, and Mycoplasma colonies were observed in the pleuropneumonia-like organisms agar culture. Based on the results of the M-PCR method, 61 specimens (out of 125 samples) were scored positive for Mycoplasma presence. Furthermore, 20 samples were positive according to the P30-PCR data. It should be mentioned that the MA-PCR was performed based on the P80 gene on 125 total samples to furtherverify the results for M.agalactiae detection. Based on the obtained data, P30 and P80 genes were presented and amplified in all Iranian M. agalactiae isolates (n=20). Our results indicated that the P30 gene was conserved and specific to all Iranian M. agalactiae isolates and this new P30-PCR method (as an MA-PCR technique) might be useful in the detection of this pathogen.
Topics: Animals; Goat Diseases; Iran; Mycoplasma; Mycoplasma Infections; Mycoplasma agalactiae; RNA, Ribosomal, 16S; Sheep
PubMed: 33818958
DOI: 10.22092/ari.2020.115005.1141 -
Journal of Medical Microbiology Jan 2022is a bacterium belonging to the class . It causes acute and chronic infections of the urogenital tract. The main features of this bacterium are an absence of cell wall...
is a bacterium belonging to the class . It causes acute and chronic infections of the urogenital tract. The main features of this bacterium are an absence of cell wall and a reduced genome size (517-622 protein-encoding genes). Previously, we have isolated morphologically unknown colonies called micro-colonies (MCs) from the serum of patients with inflammatory urogenital tract infection. MCs are functionally different from the typical colonies (TCs) in terms of metabolism and cell division. To determine the physiological differences between MCs and TCs of and elucidate the pathways of formation and growth of MCs by a comparative proteomic analysis of these two morphological forms. LC-MS proteomic analysis of TCs and MCs using an Ultimate 3000 RSLC nanoHPLC system connected to a QExactive Plus mass spectrometer. The study of the proteomic profiles of colonies allowed us to reconstruct their energy metabolism pathways. In addition to the already known pentose phosphate and arginine deamination pathways, can utilise ribose phosphate and deoxyribose phosphate formed by nucleoside catabolism as energy sources. Comparative proteomic HPLC-MS analysis revealed that the proteomic profiles of TCs and MCs were different. We assume that MC cells preferably utilised deoxyribonucleosides, particularly thymidine, as an energy source rather than arginine or ribonucleosides. Utilisation of deoxyribonucleosides is less efficient as compared with that of ribonucleosides and arginine in terms of energy production. Thymidine phosphorylase DeoA is one of the key enzymes of deoxyribonucleosides utilisation. We obtained a DeoA overexpressing mutant that exhibited a phenotype similar to that of MCs, which confirmed our hypothesis. In addition to the two known pathways for energy production (arginine deamination and the pentose phosphate pathway) can use deoxyribonucleosides and ribonucleosides. MC cells demonstrate a reorganisation of energy metabolism: unlike TC cells, they preferably utilise deoxyribonucleosides, particularly thymidine, as an energy source rather than arginine or ribonucleosides. Thus MC cells enter a state of energy starvation, which helps them to survive under stress, and in particular, to be resistant to antibiotics.
Topics: Arginine; Humans; Mycoplasma Infections; Mycoplasma hominis; Phenotype; Phosphates; Proteome; Ribonucleosides; Thymidine
PubMed: 35037614
DOI: 10.1099/jmm.0.001468 -
Cell Reports Jun 2020Here, we propose an approach to identify active metabolic pathways by integrating gene essentiality analysis and protein abundance. We use two bacterial species...
Here, we propose an approach to identify active metabolic pathways by integrating gene essentiality analysis and protein abundance. We use two bacterial species (Mycoplasma pneumoniae and Mycoplasma agalactiae) that share a high gene content similarity yet show significant metabolic differences. First, we build detailed metabolic maps of their carbon metabolism, the most striking difference being the absence of two key enzymes for glucose metabolism in M. agalactiae. We then determine carbon sources that allow growth in M. agalactiae, and we introduce glucose-dependent growth to show the functionality of its remaining glycolytic enzymes. By analyzing gene essentiality and performing quantitative proteomics, we can predict the active metabolic pathways connected to carbon metabolism and show significant differences in use and direction of key pathways despite sharing the large majority of genes. Gene essentiality combined with quantitative proteomics and metabolic maps can be used to determine activity and directionality of metabolic pathways.
Topics: Bacterial Proteins; Carbon; Chromatography, High Pressure Liquid; Genes, Essential; Glucose; Glycolysis; Mass Spectrometry; Metabolic Networks and Pathways; Mycoplasma agalactiae; Mycoplasma pneumoniae; Proteome; Proteomics
PubMed: 32492430
DOI: 10.1016/j.celrep.2020.107722 -
BMC Veterinary Research Oct 2021Mycoplasma species have been associated with economically important diseases affecting ruminants worldwide and include contagious bovine pleuropneumonia (CBPP),...
BACKGROUND
Mycoplasma species have been associated with economically important diseases affecting ruminants worldwide and include contagious bovine pleuropneumonia (CBPP), contagious caprine pleuropneumonia (CCPP) and contagious agalactia, listed by the World Organisation for Animal Health (OIE). The Mycoplasma Team at the Animal and Plant Health Agency provides an identification service for Mycoplasma and Ureaplasma species of veterinary importance to the United Kingdom (UK), supporting the detection of new and emerging pathogens, as well as contributing to the surveillance of endemic, and the OIE listed diseases exotic to the UK. Mycoplasma and other Mollicutes species were identified from diagnostic samples from farmed ruminants in England and Wales using a combination of culture and 16S rRNA gene-based PCR-denaturing gradient gel electrophoresis, submitted between 2005 and 2019.
RESULTS
A total of 5578 mollicutes identifications, which include mycoplasmas and the related acholeoplasmas and ureaplasmas, were made from farmed ruminant animals during the study period. Throughout the study period, the pathogen Mycoplasma bovis was consistently the most frequently identified species, accounting for 1411 (32%) of 4447 molecular identifications in cattle, primarily detected in the lungs of pneumonic calves, followed by joints and milk of cattle showing signs of arthritis and mastitis, respectively. M. bovirhinis, M. alkalescens, M. dispar, M. arginini and Ureaplasma diversum, were also common. Mixed species, principally M. bovis with M. alkalescens, M. arginini or M. bovirhinis were also prevalent, particularly from respiratory samples. The non-cultivable blood-borne haemoplasmas Candidatus 'Mycoplasma haemobos' and Mycoplasma wenyonii were identified from cattle, with the latter species most often associated with milk-drop. M. ovipneumoniae was the predominant species identified from sheep and goats experiencing respiratory disease, while M. conjunctivae preponderated in ocular samples. The UK remains free of the ruminant mycoplasmas listed by OIE.
CONCLUSIONS
The continued high prevalence of M. bovis identifications confirms its ongoing dominance and importance as a significant pathogen of cattle in England and Wales, particularly in association with respiratory disease. M. ovipneumoniae has seen a general increase in prevalence in recent years, notably in coughing lambs and should therefore be considered as a primary differential diagnosis of respiratory disease in small ruminants.
Topics: Animal Diseases; Animals; England; Mycoplasma; Mycoplasma Infections; RNA, Ribosomal, 16S; Ruminants; Tenericutes; Wales
PubMed: 34641885
DOI: 10.1186/s12917-021-03037-y -
Applied and Environmental Microbiology May 2021MalF has been shown to be required for virulence in the important avian pathogen To characterize the function of MalF, predicted to be part of a putative ABC...
MalF has been shown to be required for virulence in the important avian pathogen To characterize the function of MalF, predicted to be part of a putative ABC transporter, we compared metabolite profiles of a mutant with a transposon inserted in (MalF-deficient ST mutant 04-1; Δ) with those of wild-type bacteria using gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry. Of the substrates likely to be transported by an ABC transport system, glycerol was detected at significantly lower abundance in the Δ mutant, compared to the wild type. Stable isotope labeling using [U-C]glycerol and reverse transcription-quantitative PCR analysis indicated that MalF was responsible for the import of glycerol into and that, in the absence of MalF, the transcription of , which encodes a second transporter, GtsA, was upregulated, potentially to increase the import of glycerol-3-phosphate into the cell to compensate for the loss of MalF. The loss of MalF appeared to have a global effect on glycerol metabolism, suggesting that it may also play a regulatory role, and cellular morphology was also affected, indicating that the change to glycerol metabolism may have a broader effect on cellular organization. Overall, this study suggests that the reduced virulence of the Δ mutant is due to perturbed glycerol uptake and metabolism and that the operon including should be reannotated as to reflect its function in glycerol transport. Many mycoplasmas are pathogenic and cause disease in humans and animals. causes chronic respiratory disease in chickens and infectious sinusitis in turkeys, resulting in economic losses in poultry industries throughout the world. Expanding our knowledge about the pathogenesis of mycoplasma infections requires better understanding of the specific gene functions of these bacteria. In this study, we have characterized the metabolic function of a protein involved in the pathogenicity of , as well as its effect on expression of selected genes, cell phenotype, and HO production. This study is a key step forward in elucidating why this protein plays a key role in virulence in chickens. This study also emphasizes the importance of functional characterization of mycoplasma proteins, using tools such as metabolomics, since prediction of function based on homology to other bacterial proteins is not always accurate.
Topics: ATP-Binding Cassette Transporters; Bacterial Proteins; Chromatography, High Pressure Liquid; DNA Transposable Elements; Gas Chromatography-Mass Spectrometry; Glycerol; Hydrogen Peroxide; Mass Spectrometry; Mycoplasma gallisepticum; Reverse Transcriptase Polymerase Chain Reaction; Virulence
PubMed: 33741628
DOI: 10.1128/AEM.03112-20 -
Archives of Razi Institute Apr 2023is unique among prokaryotes because of its small size, small genomes, and complete lack of cell walls, which makes them cell wall-less prokaryotes. This study aimed to...
is unique among prokaryotes because of its small size, small genomes, and complete lack of cell walls, which makes them cell wall-less prokaryotes. This study aimed to evaluate the effect of vaccinating one-day-old chicks with inactivated and live vaccines (CRDF) of (MG) on their humoral immune response and immune organs. The Enzyme-Linked Immunosorbent Assay was used to measure Ab titers and investigate histopathological changes. A total of 130 one-day-old broiler chicks were randomly divided into four groups of 30. The groups were treated as follows: G1 included the chicks vaccinated with live F-strain MG vaccine (on eye drop of 0.03ml/dose), G2 included the chicks vaccinated with inactivated MG (0.3 ml s.c) vaccine, G3 included the chicks vaccinated with inactivated and live MG vaccines, and G4 was considered the control group, in which the chicks were not vaccinated. Blood samples were collected on days 21 and 35 of the chick's life to measure the titers of specific antibodies. On day 35, the chicks were dissected, and the bursa of Fabricius, as well as the spleen, were removed for histological evaluations. On day 21, the results showed a significant difference (≤0.05) between all vaccinated groups in Ab titers, compared to G4, with the highest mean in G3, followed by G2 and G1, in descending order. On day 35, there was a significant difference (≤0.05) between G3 and other vaccinated groups (G2 and G1), as well as G4. In addition, there was a significant increase in all vaccinated groups on day 35, compared to day 21. In G1, histopathological examination results showed a moderate lymphocytic hyperplasia bursal follicle. In G2, varying degrees of lymphoproliferative were observed in the major bursal follicle, and in G3, a marked lymphocytic hyperplasia bursal follicle was observed. In G4, on the other hand, no obvious histopathological findings were recorded. The results of the spleen histopathological evaluation showed various degrees of lymphoproliferative and moderate neutrophilic infiltrate in the red pulp in G1, and mild sinus congestion with scattered lymphocytes was recorded in the lumen in G2. In the spleen of the chicks in G3, reactive lymphoid hyperplasia was observed. In contrast to the groups mentioned above, in G4, the spleen structure showed a typical structure. It was concluded that the chicks vaccinated with inactivated and live MG vaccines experienced increased production of Ab titers and the immune stimulation of immune organs.
Topics: Animals; Chickens; Mycoplasma gallisepticum; Viral Vaccines; Hyperplasia; Immunity, Humoral
PubMed: 37396742
DOI: 10.22092/ARI.2022.359729.2464 -
Brazilian Journal of Biology = Revista... 2021Contamination of primary and cell cultures by mycoplasmas is one of the main economic and biological pitfalls in basic research, diagnosis and manufacture of...
Contamination of primary and cell cultures by mycoplasmas is one of the main economic and biological pitfalls in basic research, diagnosis and manufacture of biotechnological products. It is a common issue which may be difficult to conduct surveillance on. Mycoplasma presence may affect several physiological parameters of the cell, besides being considered an important source of inaccurate and/or non-reproducible scientific results. Each cell type presents characteristical symptoms, mainly morphological, that indicate a contamination by mycoplasma. HEp-2 cells originate from carcinoma of the larynx and are, therefore, part of the respiratory tract, which is one of mycoplasma habitats. Despite the importance these cells in several biological research (evaluation of cell proliferation and migration, apoptosis, antiviral and antitumor compounds), the alterations induced by mycoplasma contamination in HEp-2 cells have not yet been described. Here, we describe the progressive morphological alterations in culture of HEp-2 cells infected with mycoplasma, as well as the-diagnosis of the infection and its treatment. Mycoplasma contamination described within this work led to cytoplasm elongation, cell-to-cell spacing, thin plasma membrane projections, cytoplasmic vacuoles, fusion with neighboring cells, and, finally, cell death. Contamination was detected by fluorescence imaging (DAPI) and PCR reactions. The cultures were treated with BM-Cyclin antibiotic to eliminate contamination. The data presented here will be of relevance to researchers whose investigations involve cell culture, especially respiratory and HEp-2 cells.
Topics: Anti-Bacterial Agents; Cell Culture Techniques; Mycoplasma; Polymerase Chain Reaction
PubMed: 32321065
DOI: 10.1590/1519-6984.215721 -
Antimicrobial Agents and Chemotherapy May 2020Antibiotic resistance is a global concern; however, data on antibiotic-resistant spp. and are limited in comparison to similar data on other microbes. A total of 492...
Antibiotic resistance is a global concern; however, data on antibiotic-resistant spp. and are limited in comparison to similar data on other microbes. A total of 492 spp. and 13 strains obtained in Hangzhou, China, in 2018 were subjected to antimicrobial susceptibility testing for levofloxacin, moxifloxacin, erythromycin, clindamycin, and doxycycline using the broth microdilution method. The mechanisms underlying quinolone and macrolide resistance were determined. Meanwhile, a model of the topoisomerase IV complex bound to levofloxacin in wild-type spp. was built to study the quinolone resistance mutations. For spp., the levofloxacin, moxifloxacin, and erythromycin resistance rates were 84.69%, 51.44%, and 3.59% in and 82.43%, 62.16%, and 5.40% in , respectively. Of the 13 strains, 11 were resistant to both levofloxacin and moxifloxacin, and five strains showed clindamycin resistance. ParC S83L was the most prevalent mutation in levofloxacin-resistant strains, followed by ParE R448K. The two mutations GyrA S153L and ParC S91I were commonly identified in quinolone-resistant A molecular dynamics-refined structure revealed that quinolone resistance-associated mutations inhibited the interaction and reduced affinity with gyrase or topoisomerase IV and quinolones. The novel mutations S21A in the L4 protein and G2654T and T2245C in 23S rRNA and the gene were identified in erythromycin-resistant spp. As fluoroquinolone resistance in spp. and remains high in China, the rational use of antibiotics needs to be further enhanced.
Topics: Anti-Bacterial Agents; China; Drug Resistance, Bacterial; Humans; Macrolides; Microbial Sensitivity Tests; Mycoplasma Infections; Mycoplasma hominis; Quinolones; Ureaplasma; Ureaplasma Infections; Ureaplasma urealyticum
PubMed: 32229497
DOI: 10.1128/AAC.02560-19 -
Medicina (Kaunas, Lithuania) Feb 2021Ureaplasma urealyticum (UU) and Mycoplasma hominis (MH) are two commensal microorganisms that form the urogenital microbiota. Under a state of dysbiosis, both bacteria...
Ureaplasma urealyticum (UU) and Mycoplasma hominis (MH) are two commensal microorganisms that form the urogenital microbiota. Under a state of dysbiosis, both bacteria cause intrauterine infection. Therefore, the purpose of the present study was to analyze the prevalence of UU and MH among four hundred and eleven infertile women. Women between thirty and thirty-five years old were the most affected group, followed by those that were 25 and 30 years old, respectively. Cumulatively, the prevalence of single UU and MH, and coinfection, was 28.46% ( = 117), ( = 2) 0.48%, and 2.91% ( = 12), respectively, with an overall detection rate of 31.87% ( = 131). To assess the associated drug susceptibility, endocervical samples were unequally sent to Regina Maria ( = 281) and Synevo ( = 130) laboratories for further analyses. Pristinamycin (100% vs. 100%) and Josamycin (100% vs. 98.00%) were the most efficient antibiotics in eradicating UU and MH, several others also displaying a high efficiency, among which can be mentioned Doxycycline (98.23%), Minocycline (96.00%), Tetracycline (96.48% vs. 68.00%), and Erythromycin (70.17% vs. 92.00%). Based on antibiograms, Clarithromycin (88.00%), Roxithromycin (88.00%), Levofloxacin (82.00%), and Azithromycin (78.94%) can be further used in treating such infections. On the other hand, Clindamycin (4.00%) and Ciprofloxacin (12.27% vs. 2.00%) are no longer viable because both UU and MH display an intermediate response towards gained resistance. Interestingly, the efficiency of Ofloxacin (22.79% vs. 60.00%) was conflicting, this possibly suggesting a transient stage to a gradual adaptability of these microorganisms to Ofloxacin. The most susceptible age groups in each case were women that were between twenty and forty years old. It can be concluded that four antibiotics can be safely used for treating UU, MH, or dual infections whose efficiency was over 95%.
Topics: Adult; Female; Humans; Infertility, Female; Male; Mycoplasma hominis; Prevalence; Romania; Ureaplasma Infections; Ureaplasma urealyticum; Young Adult
PubMed: 33652790
DOI: 10.3390/medicina57030211