-
International Journal of Molecular... Mar 2020Human pluripotent stem cells have the potential to change the way in which human diseases are cured. Clinical-grade human embryonic stem cells and human induced... (Review)
Review
Human pluripotent stem cells have the potential to change the way in which human diseases are cured. Clinical-grade human embryonic stem cells and human induced pluripotent stem cells have to be created according to current good manufacturing practices and regulations. Quality and safety must be of the highest importance when humans' lives are at stake. With the rising number of clinical trials, there is a need for a consensus on hPSCs characterization. Here, we summarize mandatory and 'for information only' characterization methods with release criteria for the establishment of clinical-grade hPSC lines.
Topics: Bacteria; Cell- and Tissue-Based Therapy; Endotoxins; Human Embryonic Stem Cells; Humans; Induced Pluripotent Stem Cells; Mycoplasma; Pluripotent Stem Cells; Viruses
PubMed: 32244538
DOI: 10.3390/ijms21072435 -
Frontiers in Cellular and Infection... 2023Bovine respiratory disease (BRD) is the most devastating disease affecting beef and dairy cattle producers in North America. An emerging area of interest is the... (Meta-Analysis)
Meta-Analysis
BACKGROUND
Bovine respiratory disease (BRD) is the most devastating disease affecting beef and dairy cattle producers in North America. An emerging area of interest is the respiratory microbiome's relationship with BRD. However, results regarding the effect of BRD on respiratory microbiome diversity are conflicting.
RESULTS
To examine the effect of BRD on the alpha diversity of the respiratory microbiome, a meta-analysis analyzing the relationship between the standardized mean difference (SMD) of three alpha diversity metrics (Shannon's Diversity Index (Shannon), Chao1, and Observed features (OTUs, ASVs, species, and reads) and BRD was conducted. Our multi-level model found no difference in Chao1 and Observed features SMDs between calves with BRD and controls. The Shannon SMD was significantly greater in controls compared to that in calves with BRD. Furthermore, we re-analyzed 16S amplicon sequencing data from four previously published datasets to investigate BRD's effect on individual taxa abundances. Additionally, based on Bray Curtis and Jaccard distances, health status, sampling location, and dataset were all significant sources of variation. Using a consensus approach based on RandomForest, DESeq2, and ANCOM-BC2, we identified three differentially abundant amplicon sequence variants (ASVs) within the nasal cavity, ASV5_, ASV19_, and ASV37_ However, no ASVs were differentially abundant in the other sampling locations. Moreover, based on SECOM analysis, ASV37_ had a negative relationship with ASV1_, ASV5_, and ASV4_. ASV19_ had negative relationships with ASV1_, ASV4_, ASV54_, ASV7_, and ASV8_.
CONCLUSIONS
Our results confirm a relationship between bovine respiratory disease and respiratory microbiome diversity and composition, which provide additional insight into microbial community dynamics during BRD development. Furthermore, as sampling location and sample processing (dataset) can also affect results, consideration should be taken when comparing results across studies.
Topics: Cattle; Animals; Respiratory Tract Diseases; Cattle Diseases; Clostridiales; Microbiota; Mycoplasma hyorhinis
PubMed: 37743862
DOI: 10.3389/fcimb.2023.1223090 -
Applied and Environmental Microbiology Nov 2018One hundred seventy-eight mycoplasma strains isolated from South African poultry flocks between 2003 and 2015 were identified by full-genome sequencing and phylogenetic...
One hundred seventy-eight mycoplasma strains isolated from South African poultry flocks between 2003 and 2015 were identified by full-genome sequencing and phylogenetic analysis of the 16S rRNA gene and were classified as follows: (25%), (25%), , (23%), (14%), (10%), and (3%), as well as one strain (1%). MIC testing was performed on the axenic samples, and numerous strains of each species were resistant to either chlortetracycline or tylosin or both, with variable sensitivity to enrofloxacin. The strains of all species tested remained sensitive to tiamulin, except for one sample that demonstrated intermediate sensitivity. The mutation of A to G at position 2059 (A2059G) in the 23S rRNA gene, which is associated with macrolide resistance, was found in the South African and strains, as well as a clear correlation between macrolide resistance in and and mutations G354A and G748A in the L4 ribosomal protein and 23S rRNA gene, respectively. No correlation between resistance and point mutations in the genes studied could be found for Only a few strains were resistant to enrofloxacin, apart from one strain with point mutation D420N, which has been associated with quinolone resistance, and no other known markers for quinolone resistance were found in this study. Proportionally more antimicrobial-resistant strains were detected in , , and than in and Of concern, three strains showed multidrug resistance to chlortetracycline, tylosin, and oxytetracycline. Nonpathogenic poultry species are often overlooked due to their lesser impact on poultry health and production compared to the OIE-listed pathogenic strains and The use of antimicrobials as in-feed growth promoters and for the control of mycoplasmosis is common in poultry production across the world. Here, we provide evidence that certain nonpathogenic species are acquiring multidrug resistance traits. This would have significant implications if these species, for which no vaccines are applied, are able to transfer their antibiotic resistance genes to other mycoplasmas and bacteria that may enter the human food chain.
Topics: Animals; Anti-Bacterial Agents; Chickens; Chlortetracycline; Diterpenes; Drug Resistance, Multiple, Bacterial; Microbial Sensitivity Tests; Mycoplasma; Mycoplasma Infections; Phylogeny; Poultry Diseases; South Africa; Tylosin
PubMed: 30171000
DOI: 10.1128/AEM.01660-18 -
Microbial Genomics Oct 2023is a fast-growing species isolated from wild and first described in 2013. isolates have been associated with arthritis, kerato conjunctivitis, pneumonia and...
is a fast-growing species isolated from wild and first described in 2013. isolates have been associated with arthritis, kerato conjunctivitis, pneumonia and septicemia, but were also recovered from apparently healthy animals. To better understand what defines this species, we performed a genomic survey on 14 strains collected from free-ranging or zoo-housed animals between 1987 and 2017, mostly in Europe. The average chromosome size of the strains was 1,040±0,024 kbp, with 24 % G+C and 852±31 CDS. The core genome and pan-genome of the species contained 628 and 1312 protein families, respectively. The strains displayed a relatively closed pan-genome, with many features and putative virulence factors shared with species from the cluster, including the MIB-MIP Ig cleavage system, a repertoire of DUF285 surface proteins and a complete biosynthetic pathway for galactan. genomes were found to be mostly syntenic, although repertoires of mobile genetic elements, including Mycoplasma Integrative and Conjugative Elements, insertion sequences, and a single plasmid varied. Phylogenetic- and gene content analyses confirmed that was closer to the cluster than to the ruminant species and . Ancestral genome reconstruction showed that the emergence of the species was associated with the gain of 17 gene families, some of which encode defence enzymes and surface proteins, and the loss of 25 others, some of which are involved in sugar transport and metabolism. This comparative study suggests that the cluster could be extended to include . We also find evidence that the specific organization and structure of the DnaA boxes around the of may contribute to drive the remarkable fast growth of this minimal bacterium.
Topics: Animals; Genome, Bacterial; Phylogeny; Mycoplasma mycoides; Mycoplasma; Ruminants; Genomics; Membrane Proteins
PubMed: 37823548
DOI: 10.1099/mgen.0.001112 -
Journal of Artificial Organs : the... Mar 2022A major concern in the clinical application of cell therapy is the manufacturing cost of cell products, which mainly depends on quality control. The mycoplasma test, an...
A major concern in the clinical application of cell therapy is the manufacturing cost of cell products, which mainly depends on quality control. The mycoplasma test, an important biological test in cell therapy, takes several weeks to detect a microorganism and is extremely expensive. Furthermore, the manual detection of mycoplasma from images requires high-level expertise. We hypothesized that a mycoplasma identification program using a convolutional neural network could reduce the test time and improve sensitivity. To this end, we developed a program comprising three parts (mycoplasma detection, prediction, and cell counting) that allows users to evaluate the sample and verify infected/non-infected cells identified by the program. In experiments conducted, stained DNA images of positive and negative control using mycoplasma-infected and non-infected Vero cells, respectively, were used as training data, and the program results were compared with those of conventional methods, such as manual counting based on visual observation. The minimum detectable mycoplasma contaminations for manual counting and the proposed program were 10 and 5 CFU (colony-forming unit), respectively, and the test time for manual counting was 20 times that for the proposed program. These results suggest that the proposed system can realize a low-cost and streamlined manufacturing process for cellular products in cell-based research and clinical applications.
Topics: Animals; Chlorocebus aethiops; Deep Learning; Mycoplasma; Vero Cells
PubMed: 34160717
DOI: 10.1007/s10047-021-01282-4 -
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 -
Nature Communications Apr 2022Mycoplasmas have exceptionally streamlined genomes and are strongly adapted to their many hosts, which provide them with essential nutrients. Owing to their relative...
Mycoplasmas have exceptionally streamlined genomes and are strongly adapted to their many hosts, which provide them with essential nutrients. Owing to their relative genomic simplicity, Mycoplasmas have been used to develop chassis for biotechnological applications. However, the dearth of robust and precise toolkits for genomic manipulation and tight regulation has hindered any substantial advance. Herein we describe the construction of a robust genetic toolkit for M. pneumoniae, and its successful deployment to engineer synthetic gene switches that control and limit Mycoplasma growth, for biosafety containment applications. We found these synthetic gene circuits to be stable and robust in the long-term, in the context of a minimal cell. With this work, we lay a foundation to develop viable and robust biosafety systems to exploit a synthetic Mycoplasma chassis for live attenuated vectors for therapeutic applications.
Topics: Containment of Biohazards; Genomics; Mycoplasma pneumoniae
PubMed: 35393441
DOI: 10.1038/s41467-022-29574-0 -
Archives of Razi Institute Sep 2018Mycoplasma gallisepticum and Mycoplasma synoviae are the causative agents of avian mycoplasmosis in commercial poultry. Among the available tools, polymerase chain... (Comparative Study)
Comparative Study
Mycoplasma gallisepticum and Mycoplasma synoviae are the causative agents of avian mycoplasmosis in commercial poultry. Among the available tools, polymerase chain reaction (PCR) and culture are confirmatory tools for the diagnosis of mycoplasmosis after the initial serological screening of suspected birds. Overall, 181 samples were analyzed, 152 (84%) and 103 (57%) of which were found positive by PCR and culture, respectively. Further, 54 (92%) broiler samples were found positive for general avian mycoplasma. Among the total positive samples, MS positivity was as high as 72 (47%) by PCR, while it was 45 (44%) by culture. MG positivity was 23% and 25% in PCR- and culture-positive samples. MG grows more easily compared to MS. The agreement value between the tests was 67%. Overall, flock wise prevalence was not much varied. The prevalence of mycoplasmosis was higher during winter. Our study confirmed that PCR is the most sensitive and reliable tool for the diagnosis of avian mycoplasmosis in field samples.
Topics: Animals; Chickens; Culture Techniques; Mycoplasma Infections; Mycoplasma gallisepticum; Mycoplasma synoviae; Polymerase Chain Reaction; Poultry Diseases
PubMed: 30280844
DOI: 10.22092/ari.2017.108217.1085 -
Sheng Wu Gong Cheng Xue Bao = Chinese... Dec 2023is the pathogen causing swine mycoplasmal pneumonia. The lack of well-established animal models of . infection has delayed the progress of . -related anti-infection... (Review)
Review
is the pathogen causing swine mycoplasmal pneumonia. The lack of well-established animal models of . infection has delayed the progress of . -related anti-infection immunity studies. This paper reviews the inflammatory response, the recognition of . by the innate immune system, and the role of innate immune cells, complement system, antimicrobial peptides, autophagy, and apoptosis in . infection. The aim was to elucidate the important roles played by the components of the innate immune system in the control of . infection, and prospect key research directions of innate immune response of . infection in the future.
Topics: Animals; Swine; Mycoplasma hyopneumoniae; Pneumonia of Swine, Mycoplasmal; Immunity, Innate
PubMed: 38147980
DOI: 10.13345/j.cjb.230504 -
American Journal of Transplantation :... Jun 2021Hyperammonemia syndrome (HS) is a rare complication with high mortality described after lung transplantation. Its pathophysiology is still unclear, but previous studies,...
Hyperammonemia syndrome (HS) is a rare complication with high mortality described after lung transplantation. Its pathophysiology is still unclear, but previous studies, including murine models, have linked the identification of Mycoplasmataceae in airway specimens with HS occurrence. This study explores the association between Mycoplasmataceae polymerase chain reaction (PCR) positivity, ammonia levels, HS, and mortality post-lung transplant. Adults who underwent lung transplantation between July 2017 and August 2019 had prospective surveillance testing for Mycoplasma and Ureaplasma using PCR on post-operative bronchoscopy samples. One hundred and fifty-nine patients underwent lung transplantation during the study period. Mean age was 54 (±13) years; baseline diseases were predominantly pulmonary fibrosis (37.7%) and chronic obstructive pulmonary disease (35.8%). Mycoplasma and/or Ureaplasma airway positivity was found in 42 (26.4%) of tested patients, represented mostly by M. salivarium (26/43; 60.4%), U. parvum (7/43; 16.2%), and U. urealyticum (5/43; 11.6%). Median peak ammonia levels were higher in those with Ureaplasma colonization compared to uncolonized patients (p = .04), however, only three patients developed HS. Recipient airway Ureaplasma positivity was independently associated with younger (aOR 0.94, 95% CI 0.88-0.99, p = .04) and female donors (aOR 4.29; 95% CI 1.01-18.2, p = .05).
Topics: Adult; Ammonia; Animals; Female; Humans; Lung Transplantation; Mice; Middle Aged; Mycoplasma; Prospective Studies; Ureaplasma; Ureaplasma Infections; Ureaplasma urealyticum
PubMed: 33179447
DOI: 10.1111/ajt.16394