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MSystems Apr 2024species are able to produce and release secreted proteins, such as toxins, adhesins, and virulence-related enzymes, involved in bacteria adhesion, invasion, and immune...
UNLABELLED
species are able to produce and release secreted proteins, such as toxins, adhesins, and virulence-related enzymes, involved in bacteria adhesion, invasion, and immune evasion between the pathogen and host. Here, we investigated a novel secreted protein, MbovP0725, from encoding a putative haloacid dehalogenase (HAD) hydrolase function of a key serine/threonine phosphatase depending on Mg for the dephosphorylation of its substrate and it was most active at pH 8 to 9 and temperatures around 40°C. A transposon insertion mutant strain of HB0801 that lacked the protein MbovP0725 induced a stronger inflammatory response but with a partial reduction of adhesion ability. Using transcriptome sequencing and quantitative reverse transcription polymerase chain reaction analysis, we found that the mutant was upregulated by the mRNA expression of genes from the glycolysis pathway, while downregulated by the genes enriched in ABC transporters and acetate kinase-phosphate acetyltransferase pathway. Untargeted metabolomics showed that the disruption of the gene caused the accumulation of 9-hydroxyoctadecadienoic acids and the consumption of cytidine 5'-monophosphate, uridine monophosphate, and adenosine monophosphate. Both the exogenous and endogenous MbvoP0725 protein created by purification and transfection inhibited lipopolysaccharide (LPS)-induced IL-1β, IL-6, and TNF-α mRNA production and could also attenuate the activation of MAPK-associated pathways after LPS treatment. A pull-down assay identified MAPK p38 and ERK as potential substrates for MbovP0725. These findings define metabolism- and virulence-related roles for a HAD family phosphatase and reveal its ability to inhibit the host pro-inflammatory response.
IMPORTANCE
() infection is characterized by chronic pneumonia, otitis, arthritis, and mastitis, among others, and tends to involve the suppression of the immune response via multiple strategies to avoid host cell immune clearance. This study found that MbovP0725, a haloacid dehalogenase (HAD) family phosphatase secreted by , had the ability to inhibit the host pro-inflammatory response induced by lipopolysaccharide. Transcriptomic and metabolomic analyses were used to identify MbovP0725 as an important phosphatase involved in glycolysis and nucleotide metabolism. The transposon mutant strain T8.66 lacking MbovP0725 induced a higher inflammatory response and exhibited weaker adhesion to host cells. Additionally, T8.66 attenuated the phosphorylation of MAPK P38 and ERK and interacted with the two targets. These results suggested that MbovP0725 had the virulence- and metabolism-related role of a HAD family phosphatase, performing an anti-inflammatory response during infection.
Topics: Female; Humans; Mycoplasma bovis; Lipopolysaccharides; Bacterial Adhesion; Mycoplasma Infections; Immunity; Phosphoprotein Phosphatases; RNA, Messenger; Serine
PubMed: 38440990
DOI: 10.1128/msystems.00891-23 -
Italian Journal of Pediatrics Mar 2024The prevalence of macrolide-resistant Mycoplasma pneumoniae has increased considerably. Treatment in children has become challenging. This study aimed to evaluate the...
BACKGROUND
The prevalence of macrolide-resistant Mycoplasma pneumoniae has increased considerably. Treatment in children has become challenging. This study aimed to evaluate the efficacy of doxycycline therapy for macrolide-resistant Mycoplasma pneumoniae pneumonia in children at different periods.
METHODS
We retrospectively analyzed the data of patients with macrolide-resistant Mycoplasma pneumoniae pneumonia hospitalized between May 2019 to August 2022. According to treatment, patients were divided into three groups: oral doxycycline treatment alone (DOX group), changed from intravenous azithromycin to oral doxycycline (ATD group), and intravenous azithromycin treatment alone (AZI group). ATD group cases were separated into two sub-groups: intravenous azithromycin treatment<3 days (ATD1 group) and ≥ 3 days (ATD2 group). Clinical symptoms were compared in each group and adjusted by Propensity score matching (PSM) analysis.
RESULTS
A total of 106 were recruited in this study. 17 (16%) were in DOX group, 58 (55%) in ATD group, and 31(29%) in AZI group. Compared with ATD group and AZI group, the DOX group showed shorter hospitalization duration and fever duration after treatment, while higher rate of chest radiographic improvement. After using PSM analysis, shorter days to hospitalization duration (P = 0.037) and to fever duration after treatment (P = 0.027) in DOX + ATD1 group than in ATD2 group was observed. A higher number of patients in the DOX + ATD1 group achieved defervescence within 72 h (P = 0.031), and fewer children received glucocorticoid adjuvant therapy (P = 0.002). No adverse reactions associated with doxycycline was observed during treatment.
CONCLUSIONS
Children receiving early oral doxycycline had a shorter duration of fever and hospitalization in macrolide-resistant Mycoplasma pneumoniae patients.
Topics: Child; Humans; Doxycycline; Mycoplasma pneumoniae; Macrolides; Azithromycin; Retrospective Studies; Anti-Bacterial Agents; Pneumonia, Mycoplasma
PubMed: 38439015
DOI: 10.1186/s13052-024-01615-y -
The Journal of Maternal-fetal &... Dec 2024The present study investigated the relationship between bronchopulmonary dysplasia (BPD) and intra-amniotic infection with species.
OBJECTIVES
The present study investigated the relationship between bronchopulmonary dysplasia (BPD) and intra-amniotic infection with species.
METHODS
This was a single-center, retrospective cohort study. Patients with singleton pregnancies who underwent inpatient management at our department for preterm premature rupture of membranes (PPROM), preterm labor, cervical insufficiency, and asymptomatic cervical shortening at 22-33 gestational weeks were included. Amniocentesis was indicated for patients with PPROM or an elevated maternal C-reactive protein level (≥0.58 mg/dL). Patients with an amniotic fluid IL-6 concentration ≥3.0 ng/mL were diagnosed with intra-amniotic inflammation, while those with positive aerobic, anaerobic, , and spp. cultures were diagnosed with microbial invasion of the amniotic cavity (MIAC). Patients who tested positive for both intra-amniotic inflammation and MIAC were considered to have intra-amniotic infection. An umbilical vein blood IL-6 concentration >11.0 pg/mL indicated fetal inflammatory response syndrome (FIRS). The maternal inflammatory response (MIR) and fetal inflammatory response (FIR) were staged using the Amsterdam Placental Workshop Group Consensus Statement.
RESULTS
Intra-amniotic infection with spp. was diagnosed in 37 patients, intra-amniotic infection without spp. in 28, intra-amniotic inflammation without MIAC in 58, and preterm birth without MIR/FIR and FIRS in 86 as controls. Following an adjustment for gestational age at birth, the risk of BPD was increased in patients with intra-amniotic infection with spp. (adjusted odds ratio: 10.5; 95% confidence interval: 1.55-71.2), but not in those with intra-amniotic infection without spp. or intra-amniotic inflammation without MIAC.
CONCLUSION
BPD was only associated with intra-amniotic infection with species.
Topics: Pregnancy; Infant, Newborn; Humans; Female; Ureaplasma; Chorioamnionitis; Retrospective Studies; Bronchopulmonary Dysplasia; Prevalence; Interleukin-6; Prenatal Exposure Delayed Effects; Placenta; Premature Birth; Fetal Membranes, Premature Rupture; Amniotic Fluid; Inflammation
PubMed: 38418200
DOI: 10.1080/14767058.2024.2320670 -
Emerging Microbes & Infections Dec 2024Paediatric pneumonia (MPP) is a heterogeneous disease with a diverse spectrum of clinical phenotypes. No studies have demonstrated the relationship between underlying...
Paediatric pneumonia (MPP) is a heterogeneous disease with a diverse spectrum of clinical phenotypes. No studies have demonstrated the relationship between underlying endotypes and clinical phenotypes as well as prognosis about this disease. Thus, we conducted a multicentre prospective longitudinal study on children hospitalized for MPP between June 2021 and March 2023, with the end of follow-up in August 2023. Blood samples were collected and processed at multiple time points. Multiplex cytokine assay was performed to characterize serum cytokine profiles and their dynamic changes after admission. Cluster analysis based on different clinical phenotypes was conducted. Among the included 196 patients, the levels of serum IL-17A and IL-6 showed remarkable variabilities. Four cytokine clusters based on the two cytokines and four clinical groups were identified. Significant elevation of IL-17A mainly correlated with diffuse bronchiolitis and lobar lesion by airway mucus hypersecretions, while that of IL-6 was largely associated with lobar lesion which later developed into lung necrosis. Besides, glucocorticoid therapy failed to inhibit IL-17A, and markedly elevated IL-17A and IL-6 levels may correlate with lower airway obliterans. Our study provides critical relationship between molecular signatures (endotypes) and clustered clinical phenotypes in paediatric patients with MPP.
Topics: Humans; Child; Mycoplasma pneumoniae; Interleukin-6; Interleukin-17; Prospective Studies; Longitudinal Studies; Pneumonia, Mycoplasma; Cytokines
PubMed: 38407218
DOI: 10.1080/22221751.2024.2324078 -
Theriogenology Apr 2024Mycoplasmas colonize fish, reptiles, birds and mammals, being commensals or causing diseases, sometimes severe in ruminants, swine, poultry, or wildlife animals. So far,...
Mycoplasmas colonize fish, reptiles, birds and mammals, being commensals or causing diseases, sometimes severe in ruminants, swine, poultry, or wildlife animals. So far, 15 species of canine Mycoplasma spp. have been described. Conflicting results have been presented regarding the pathogenicity of Mycoplasma spp. Although many virulence factors of these bacteria have been described, they still require attention. The main aim of our study was to evaluate the presence of known canine Mycoplasmas in the male reproductive tract of clinically healthy dogs. The second aim was to check if Mycoplasma spp. cause any abnormalities in semen quality that could have further consequences and to propose the schemes for managing the carriers. 83.3% of examined dogs were Mycoplasma spp. -positive dogs, and most of them were the carriers of more than one species. Six dogs had azoospermic ejaculates. The total spermatozoa numbers were similar in Mycoplasma -positive and negative groups. Motility was slightly higher in Mycoplasma spp.-negative group, but the difference was not statistically significant. There was no significant difference in semen characteristics between the carriers and Mycoplasma spp.-negative dogs. Neither the individual species nor the number of species strains had a significant effect on sperm morphological parameters as well as viability. Semen quality parameters are not correlated with the species found on the prepuce. Over 70% Mycoplasma spp.- positive dogs have more than one species of this bacteria. Despite finding mycoplasmas in azoospermic dogs, we suggest that they were not the cause of infertility. Mycoplasma spp. could be a part of normal microbiota in canine prepuce in individuals without any clinical signs.
Topics: Male; Dogs; Animals; Swine; Mycoplasma; Semen Analysis; Semen; Azoospermia; Spermatozoa; Mammals; Dog Diseases; Swine Diseases
PubMed: 38402701
DOI: 10.1016/j.theriogenology.2024.02.018 -
Journal of Dairy Science Jul 2024The bovine hemoplasmas include Mycoplasma wenyonii and Candidatus Mycoplasma haemobos, which are increasingly recognized as infecting cattle throughout the world....
The bovine hemoplasmas include Mycoplasma wenyonii and Candidatus Mycoplasma haemobos, which are increasingly recognized as infecting cattle throughout the world. Infection with hemotropic mycoplasma has been reported to be widespread in mature dairy cows, but little is known about its prevalence in calves and heifers. The objective of this study was to investigate the prevalence and dynamics of infection with M. wenyonii and C. M. haemobos in calves and replacement heifers on Michigan dairy farms and assess the potential associations between infection status and hematological values. The study was designed as a prospective cross-sectional study with a longitudinal component. A convenience sample of 11 farms agreed to participate and were visited twice between March and September 2022. During the first farm visit, researchers collected blood samples from up to 94 animals per farm distributed among newborn and preweaning calves (n ≤ 31), weaned calves (n = 21), pre-breeding heifers (n = 21), and pregnant heifers (n = 21). During the first visit, blood samples (n = 174) were also collected from a convenience sample of mature cows to confirm the herd infection status. The same calves and heifers were sampled again ∼95 d (±3.0) later. During the first visit, blood samples were collected from 797 calves and replacement heifers, whereas 675 samples were collected during the second visit due to the inability to locate some animals. Detection of M. wenyonii and C. M. haemobos was based on results of real-time PCR. The hematocrit was determined using microcentrifugation, and the concentration of leukocytes using an automated cell counter. In all herds, most mature cows that were sampled tested positive for infection. The within-herd apparent prevalence of hemoplasma in calves and replacement heifers was 100% for both M. wenyonii and C. M. haemobos. The apparent prevalence of hemoplasma in youngstock was associated with age. In calves that were 1 to 6 mo old, the prevalence of infection was 6% to 8% but sharply increased to 31% by 8 mo of age. In older animals, the prevalence remained high, and was almost 100% in animals greater than 17 mo of age. Based on calves and heifers sampled twice, the cumulative incidence varied widely among herds, ranging from 3.7% to 96.0%, and increased with the age of the animals. We found no difference in hematocrit or number of lymphocytes, monocytes, neutrophils, or total leukocytes based on infection status. The number of eosinophils was greater in infected animals. This is the first study to report the prevalence of hemoplasmas in calves and replacement heifers in the United States. It indicates that young calves can be infected with hemoplasmas, but the rate of infection is low. The likelihood of infection increases as animals age, with a notable rise in the proportion of infected heifers occurring by 8 mo old, and the prevalence eventually reaching nearly 100% in older animals. Once infected, heifers appear to remain chronic carriers. Hemoplasma infection alone does not usually lead to the development of clinical signs, and most of the animals remain apparently healthy.
Topics: Animals; Cattle; Female; Mycoplasma; Mycoplasma Infections; Prevalence; Cattle Diseases; Cross-Sectional Studies; Michigan; Prospective Studies; Farms
PubMed: 38395406
DOI: 10.3168/jds.2023-24395 -
High diversity, novel genotypes, and vertical transmission of hemotropic Mycoplasma in micromammals.Comparative Immunology, Microbiology... Apr 2024Hemotropic mycoplasmas (hemoplasmas) are emerging zoonotic pathogens. Micromammals have received little attention as hosts for hemoplasmas despite their ubiquitous...
Hemotropic mycoplasmas (hemoplasmas) are emerging zoonotic pathogens. Micromammals have received little attention as hosts for hemoplasmas despite their ubiquitous presence, high population abundances, and close association with humans. A PCR protocol targeting a fragment of the 16 S rRNA gene and direct sequencing in blood samples of 189 adult specimens and 35 fetuses belonging to three species of Eulipotyphla (shrews) and seven species of Rodentia, captured in three ecologically diverse habitats in North-Eastern Spain (Steppe, High Mountain, Mediterranean) yielded and occurrence of 26%, including 36% of 39 shrews and 23% of 150 rodents. Sequencing revealed the presence of 14 nucleotide sequence types (ntST) among the 56 readable sequences. In general, each ntST was associated with a given host species, although in some cases, the same ntST was sequenced in different species (chiefly rodents). Most ntST were closely related to rodent and/or bat hemoplasmas, but one was identical with Mycoplasma haemocanis/haemofelis, and others can be considered novel genotypes. High sequence diversity was detected in rodents, whereas in the white-toothed shrew (Crocidura russula), 9/11 sequences from two distant areas were identical. Phylogenetic and network analyses classified our sequences in different clades including hemoplasmas of rodents, carnivores, bats, and humans. Twelve of the fetuses (34.2%) of 9/12 litters (75.0%) of shrews and rodents were hemoplasma-positive, indicating frequent vertical transmission. Our study contributes to expanding our knowledge about the distribution, diversity, and transmission of hemoplasmas.
Topics: Animals; Humans; Mycoplasma Infections; Phylogeny; Shrews; RNA, Ribosomal, 16S; DNA, Bacterial; Mycoplasma; Carnivora; Rodentia; Genotype; Chiroptera
PubMed: 38387227
DOI: 10.1016/j.cimid.2024.102151 -
Epidemiology and Infection Feb 2024
Topics: Humans; Mycoplasma pneumoniae; Pneumonia, Mycoplasma; Disease Outbreaks
PubMed: 38380476
DOI: 10.1017/S0950268824000293 -
Journal of Structural Biology Jun 2024Cellular cryo-electron tomography (cryo-ET) has emerged as a key method to unravel the spatial and structural complexity of cells in their near-native state at...
Cellular cryo-electron tomography (cryo-ET) has emerged as a key method to unravel the spatial and structural complexity of cells in their near-native state at unprecedented molecular resolution. To enable quantitative analysis of the complex shapes and morphologies of lipid membranes, the noisy three-dimensional (3D) volumes must be segmented. Despite recent advances, this task often requires considerable user intervention to curate the resulting segmentations. Here, we present ColabSeg, a Python-based tool for processing, visualizing, editing, and fitting membrane segmentations from cryo-ET data for downstream analysis. ColabSeg makes many well-established algorithms for point-cloud processing easily available to the broad community of structural biologists for applications in cryo-ET through its graphical user interface (GUI). We demonstrate the usefulness of the tool with a range of use cases and biological examples. Finally, for a large Mycoplasma pneumoniae dataset of 50 tomograms, we show how ColabSeg enables high-throughput membrane segmentation, which can be used as valuable training data for fully automated convolutional neural network (CNN)-based segmentation.
Topics: Cryoelectron Microscopy; Electron Microscope Tomography; Software; Algorithms; Image Processing, Computer-Assisted; Neural Networks, Computer; Cell Membrane; Mycoplasma pneumoniae; User-Computer Interface; Imaging, Three-Dimensional
PubMed: 38367824
DOI: 10.1016/j.jsb.2024.108067 -
Vaccine Mar 2024Vaccination is the most cost-effective tool to control contagious bovine pleuropneumonia. The vaccines currently used in Africa are derived from a live strain called T1,...
Vaccination is the most cost-effective tool to control contagious bovine pleuropneumonia. The vaccines currently used in Africa are derived from a live strain called T1, which was attenuated by passage in embryonated eggs and broth culture. The number of passages is directly correlated to the degree of attenuation of the vaccinal strains and inversely correlated to their immunogenicity in cattle. Current quality control protocols applied to vaccine batches allow the assessment of identity, purity, and titers, but cannot assess the level of genetic drift form the parental vaccine strains. Deep sequencing was used to assess the genetic drift generated over controlled in vitro passages of the parental strain, as well as on commercial vaccine batches. Signatures of cloning procedures were detected in some batches, which imply a deviation from the standard production protocol. Deep sequencing is proposed as a new tool for the identity and stability control of T1 vaccines.
Topics: Animals; Cattle; Bacterial Vaccines; Pleuropneumonia; Africa; Vaccines, Attenuated; Quality Control; High-Throughput Nucleotide Sequencing; Pleuropneumonia, Contagious; Cattle Diseases; Mycoplasma mycoides
PubMed: 38365481
DOI: 10.1016/j.vaccine.2024.02.031