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Frontiers in Microbiology 2022is a common opportunistic zoonotic pathogen, and its ongoing acquisition of antimicrobial resistance genes poses challenges to clinical treatments. Human-sourced whole...
is a common opportunistic zoonotic pathogen, and its ongoing acquisition of antimicrobial resistance genes poses challenges to clinical treatments. Human-sourced whole genomic sequencing of human isolates has been reported, but pig-sourced isolates have not been thoroughly investigated even though these animals can serve as reservoirs for human infections. In the current study, we report a molecular epidemiological investigation to unravel the antimicrobial and virulence gene risk factors for contamination in 9 pig farms in 3 different cities in Zhejiang Province, China. We collected 541 swab samples from healthy pigs and 30 were confirmed as . All 30 isolates were resistant to tetracyclines, macrolides, sulfonamides, β-lactams and chloramphenicol, and all were multiple drug-resistant and 27 were strong biofilm formers. Phylogenetic analyses indicated these 30 isolates clustered together in 2 major groups. Whole genome sequencing demonstrated that the isolates possessed 91 different antimicrobial resistance genes belonging to 30 antimicrobial classes including and All isolates contained mobile genetic elements including integrative conjugative elements (ICEs) and integrative and mobilizable elements (IMEs). Minimum inhibitory concentration (MIC) testing indicated direct correlates between cognate genes and antimicrobial resistance. We also identified 95 virulence factors, almost all isolates contained 20 fimbrial and flagellar operons, and this represents the greatest number of these operon types found in a single species among all sequenced bacterial genomes. These genes regulate biofilm formation and represent a confounding variable for treating infections. Our isolates were present in healthy animals, and multiple drug resistance in these isolates may serve as a reservoir for other intestinal and environmental Enterobacteriaceae members. This prompts us to more strictly regulate veterinary antibiotic use.
PubMed: 35875581
DOI: 10.3389/fmicb.2022.952982 -
Nan Fang Yi Ke Da Xue Xue Bao = Journal... May 2023To construct a gene knockout strain of and explore the effect of gene deletion on biological characteristics of .
OBJECTIVE
To construct a gene knockout strain of and explore the effect of gene deletion on biological characteristics of .
METHODS
Fusion PCR was used to obtain the fusion gene of and the kanamycin-resistant gene , which was ligated with the suicide vector pCVD442 and transduced into . The gene knockout strain of was obtained after homologous recombination with the suicide vector. PCR and Sanger sequencing were used to identify genomic deletion of gene in the genetically modified strain. The concentration of molybdate in the wild-type and gene knockout strains was determined using inductively coupled plasma mass spectrometry (ICP-MS), and their survival ability in LB medium was compared under both aerobic and anaerobic conditions.
RESULTS
PCR and sanger sequencing confirmed genomic deletion of gene in the obtained strain. The concentration of intracellular molybdenum in the gene knockout strain was 1.22 mg/kg, significantly lower than that in the wild-type strain (1.46 mg/kg, < 0.001). Under the aerobic condition, the gene knockout strain grown in LB medium showed no significant changes in survival ability compared with the wild-type strain, but its proliferation rate decreased significantly under the anaerobic condition and also when cultured in nitrate-containing LB medium under anaerobic condition.
CONCLUSION
Homologous recombination with the suicide vector can be used for gene knockout in . gene participates in molybdate uptake and is associated with anaerobic growth of Proteus mirabilis in the presence of nitrate.
Topics: Humans; Gene Deletion; Nitrates; Proteus mirabilis; Gene Knockout Techniques
PubMed: 37313829
DOI: 10.12122/j.issn.1673-4254.2023.05.23 -
Frontiers in Immunology 2020Bacterial DNAs are constantly detected in atherosclerotic plaques (APs), suggesting that a combination of chronic infection and inflammation may have roles in AP...
Bacterial DNAs are constantly detected in atherosclerotic plaques (APs), suggesting that a combination of chronic infection and inflammation may have roles in AP formation. A series of studies suggested that certain Gram-negative bacteria were able to interact with dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin [DC-SIGN; cluster of differentiation (CD) 209] or langerin (CD207), thereby resulting in deposition of CD209s at infection sites. We wondered if (a member of Proteobacteria family) could interact with APs through CD209/CD207. In this study, we first demonstrated that CD209/CD207 were also receptors for that mediated adherence and phagocytosis by macrophages. interacted with fresh and CD209s/CD207-expressing APs cut from human coronary arteries, rather than in healthy and smooth arteries. These interactions were inhibited by addition of a ligand-mimic oligosaccharide and the coverage of the ligand, as well as by anti-CD209 antibody. Finally, the hearts from an atherosclerotic mouse model contained higher numbers of than that of control mice during infection-challenging. We therefore concluded that the interacts with APs in human coronary arteries CD209s/CD207. It may be possible to slow down the progress of atherosclerosis by blocking the interactions between CD209s/CD207 and certain atherosclerosis-involved bacteria with ligand-mimic oligosaccharides.
Topics: Adult; Aged; Animals; Antibodies, Monoclonal; Antigens, CD; Bacterial Adhesion; CHO Cells; Cell Adhesion Molecules; Coronary Artery Disease; Coronary Vessels; Cricetulus; Disease Models, Animal; Female; Host-Pathogen Interactions; Humans; Lectins, C-Type; Ligands; Macrophages; Male; Mannose-Binding Lectins; Mice; Mice, Inbred C57BL; Mice, Knockout, ApoE; Middle Aged; Oligosaccharides; Plaque, Atherosclerotic; Proteus mirabilis; RAW 264.7 Cells; Receptors, Cell Surface
PubMed: 33488579
DOI: 10.3389/fimmu.2020.579010 -
Microbiology Spectrum Dec 2022Ordered transposon libraries are a valuable resource for many bacterial species, especially those with difficult methods for generating targeted genetic mutations. Here,...
Ordered transposon libraries are a valuable resource for many bacterial species, especially those with difficult methods for generating targeted genetic mutations. Here, we present the construction of an ordered transposon library for the bacterial urinary tract pathogen Proteus mirabilis strain HI4320. This library will facilitate future studies into P. mirabilis biology. For large experimental screens, it may be used to overcome bottleneck constraints and avoid biased outcomes resulting from gene length. For smaller studies, the library allows sidestepping the laborious construction of single targeted mutants. This library, containing 18,432 wells, was condensed into a smaller library containing 1,728 mutants. Each selected mutant had a single transposon insertion in an open reading frame, covering 45% of predicted genes encoded by P. mirabilis HI4320. This coverage was lower than expected and was due both to library wells with no mapped insertions and a surprisingly high proportion of mixed clones and multiple transposon insertion events. We offer recommendations for improving future library construction and suggestions for how to use this P. mirabilis library resource. Ordered libraries facilitate large genetic screens by guaranteeing high genomic coverage with a minimal number of mutants, and they can save time and effort by reducing the need to construct targeted mutations. This resource is now available for P. mirabilis, a common and complicating agent of catheter-associated urinary tract infection. We also present obstacles encountered during library construction with the goal to aid others who would like to construct ordered transposon libraries in other species.
Topics: Humans; DNA Transposable Elements; Proteus mirabilis; Urinary Tract Infections; Gene Library; Urinary Tract; Proteus Infections
PubMed: 36377916
DOI: 10.1128/spectrum.03142-22 -
Antibiotics (Basel, Switzerland) Jul 2020Chaperone-usher fimbrial adhesins are powerful weapons against the uropathogens that allow the establishment of urinary tract infections (UTIs). As the antibiotic... (Review)
Review
Chaperone-usher fimbrial adhesins are powerful weapons against the uropathogens that allow the establishment of urinary tract infections (UTIs). As the antibiotic therapeutic strategy has become less effective in the treatment of uropathogen-related UTIs, the anti-adhesive molecules active against fimbrial adhesins, key determinants of urovirulence, are attractive alternatives. The best-characterized bacterial adhesin is FimH, produced by uropathogenic (UPEC). Hence, a number of high-affinity mono- and polyvalent mannose-based FimH antagonists, characterized by different bioavailabilities, have been reported. Given that antagonist affinities are firmly associated with the functional heterogeneities of different FimH variants, several FimH inhibitors have been developed using ligand-drug discovery strategies to generate high-affinity molecules for successful anti-adhesion therapy. As clinical trials have shown d-mannose's efficacy in UTIs prevention, it is supposed that mannosides could be a first-in-class strategy not only for UTIs, but also to combat other Gram-negative bacterial infections. Therefore, the current review discusses valuable and effective FimH anti-adhesive molecules active against UTIs, from design and synthesis to in vitro and in vivo evaluations.
PubMed: 32664222
DOI: 10.3390/antibiotics9070397 -
Veterinary Medicine and Science Jul 2023Pericarditis means inflammation in the pericardial sac. Pericarditis is divided into three categories based on morphology, including fibrinous, purulent, and...
Pericarditis means inflammation in the pericardial sac. Pericarditis is divided into three categories based on morphology, including fibrinous, purulent, and constrictive. In the present study, a 7-month-old male Ghezel breed sheep was examined for low weighting rate for three months. Tachypnea, tachycardia, heart friction sound, absence of fever and normal appetite were recorded in the clinical examination. In the patient's history, there was a history of perforated chest trauma behind the left scapula about three months ago. After the echocardiography examination and bacteriology procedures, purulent pericarditis caused by Proteus mirabilis was diagnosed. The bacterium was analysed using genome sequencing and new strain called Abhar114 was diagnosed. This is the first report of pericarditis caused by Proteus mirabilis in sheep.
Topics: Male; Animals; Sheep; Proteus mirabilis; Pericarditis; Echocardiography; Sheep Diseases
PubMed: 37192480
DOI: 10.1002/vms3.1160 -
Virus Research Mar 2023The crystalline formation of biofilms by Proteus blocks the urine flow which often complicates the health care of catheterized patients. Bacteriophages has been...
The crystalline formation of biofilms by Proteus blocks the urine flow which often complicates the health care of catheterized patients. Bacteriophages has been highlighted as a promising tool to control biofilm-mediated bacterial infections. Here, we isolated and characterized two newly isolated lytic phages capable of infecting clinical isolates of P. mirabilis and P. vulgaris. Moreover, insights regarding the biological and molecular characterization were analysed. Both RP6 and RP7 phages showed a Proteus-genus-specific profile, administering no lytic activity against other family of Enterobacteriaceae. The optimal MOI value of the RP6 and RP7 phages were determined as 0.1 and 0.01, respectively. The one-step growth curve showed that RP6 and RP7 phages have a short latent period of 20 min and large burst size of 220-371 PFU/ML per infected host cell. Bacteria growth was reduced immediately after the phages were added, which is shown by the optical density (OD) measurement after 24 hr. Proteus phage RP6 and RP7 were found to eradicate both the planktonic and mature biofilms produced by the Proteus isolates tested. Genome sequence of Proteus phage RP6 was found to be 58,619 bp, and a G-C content of 47%. Also, Proteus phage RP7 genome size was 103,593 bp with G-C ratio of 38.45%. A total of 70 and 172 open reading frame (ORF) was encoded in RP6 and RP7 phage genomes, respectively. Interestingly, there were no tRNA encoded by Proteus phage RP6 genome even though there is a significant G-C content difference between the phage and its host. Additionally, the exhibition of highly lytic activity and absence of virulence and antibiotic-resistant genes in both Proteus RP6 and RP7 phages emphasized that this newly isolated phages are promising for potential therapeutic phages.
Topics: Humans; Bacteriophages; Sewage; Proteus; Proteus mirabilis; Genome, Viral; Biofilms; Hospitals
PubMed: 36717023
DOI: 10.1016/j.virusres.2023.199049 -
Journal of Global Antimicrobial... Dec 2020Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis are urinary tract infection (UTI) pathogens and extended spectrum β-lactamase (ESBL)-producing pathogens...
OBJECTIVES
Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis are urinary tract infection (UTI) pathogens and extended spectrum β-lactamase (ESBL)-producing pathogens exhibit co-resistance to oral fluoroquinolones (FQ) and trimethoprim-sulphamethoxazole (TMP-SMX). This study assessed the prevalence of ESBL phenotypes and co-resistance to FQ and TMP-SMX.
METHODS
In total, 766 E. coli, 260 K. pneumoniae and 104 P. mirabilis from UTIs in 18 countries were evaluated for susceptibility in the SENTRY surveillance programme, and results interpreted using EUCAST criteria.
RESULTS
E. coli, K. pneumoniae and P. mirabilis accounted for 57.1%, 11.3% and 7.8%, respectively, of the isolates. Among E. coli, resistance to levofloxacin and TMP-SMX ranged from 21.8% to 32.7% for all isolates increasing to 66.5-67.0% among those with a ESBL phenotype (17.9% of all UTI E. coli from Europe were ESBL phenotypes). In contrast, all E. coli were susceptible to meropenem. For K. pneumoniae, resistance rates for levofloxacin and TMP-SMX were 32.2-40.0% increasing to 69.1-78.6% for ESBL phenotypes. Meropenem was the most active agent, with 7.7% resistance. Among P. mirabilis resistance to levofloxacin and TMP-SMX was 26-38.5% and increased to 100% for ESBL phenotypes. No meropenem-resistant P. mirabilis were reported.
CONCLUSIONS
High co-resistance rates were observed for oral antibiotics among ESBL phenotypes raising concerns regarding empiric use of FQ and TMP-SMX for treating resistant UTIs outside of the hospital. In contrast, intravenous carbapenems retain activity against resistant UTI pathogens. New oral options with the spectrum of the carbapenems would address an unmet need for managing resistant UTIs.
Topics: Escherichia coli; Europe; Humans; Microbial Sensitivity Tests; Urinary Tract Infections
PubMed: 33212286
DOI: 10.1016/j.jgar.2020.10.020 -
Scientific Reports Jan 2021Modification of outer membrane proteins (OMPs) is the first line of Gram-negative bacteria defence against antimicrobials. Here we point to Proteus mirabilis OMPs and...
Modification of outer membrane proteins (OMPs) is the first line of Gram-negative bacteria defence against antimicrobials. Here we point to Proteus mirabilis OMPs and their role in antibiotic and phage resistance. Protein profiles of amikacin (AMKrsv), phage (Brsv) and amikacin/phage (AMK/Brsv) resistant variants of P. mirabilis were compared to that obtained for a wild strain. In resistant variants there were identified 14, 1, 5 overexpressed and 13, 5, 1 downregulated proteins for AMKrsv, Brsv and AMK/Brsv, respectively. Application of phages with amikacin led to reducing the number of up- and downregulated proteins compared to single antibiotic treatment. Proteins isolated in AMKrsv are involved in protein biosynthesis, transcription and signal transduction, which correspond to well-known mechanisms of bacteria resistance to aminoglycosides. In isolated OMPs several cytoplasmic proteins, important in antibiotic resistance, were identified, probably as a result of environmental stress, e.g. elongation factor Tu, asparaginyl-tRNA and aspartyl-tRNA synthetases. In Brsv there were identified: NusA and dynamin superfamily protein which could play a role in bacteriophage resistance. In the resistant variants proteins associated with resistance mechanisms occurring in biofilm, e.g. polyphosphate kinase, flagella basal body rod protein were detected. These results indicate proteins important in the development of P. mirabilis antibiofilm therapies.
Topics: Amikacin; Anti-Bacterial Agents; Bacterial Infections; Bacterial Outer Membrane Proteins; Bacteriophages; Biofilms; Drug Resistance, Microbial; Gram-Negative Bacteria; Membrane Proteins; Proteus mirabilis
PubMed: 33452316
DOI: 10.1038/s41598-020-80907-9 -
Journal of Bacteriology Jan 2022Cells can use self recognition to achieve cooperative behaviors. Self-recognition genes are thought to principally evolve in tandem with partner self-recognition...
Cells can use self recognition to achieve cooperative behaviors. Self-recognition genes are thought to principally evolve in tandem with partner self-recognition alleles. However, other constraints on protein evolution could exist. Here, we have identified an interaction outside self-recognition loci that could constrain the sequence variation of a self-recognition protein. We show that during collective swarm expansion in Proteus mirabilis, self-recognition signaling co-opts SdaC, a serine transporter. Serine uptake is crucial for bacterial survival and colonization. Single-residue variants of SdaC reveal that self recognition requires an open conformation of the protein; serine transport is dispensable. A distant ortholog from Escherichia coli is sufficient for self recognition; however, a paralogous serine transporter, YhaO, is not. Thus, SdaC couples self recognition and serine transport, likely through a shared molecular interface. Self-recognition proteins may follow the framework of a complex interaction network rather than an isolated two-protein system. Understanding the molecular and ecological constraints on self-recognition proteins lays the groundwork for insights into the evolution of self recognition and emergent collective behaviors. Bacteria can receive secret messages from kin during migration. For Proteus mirabilis, these messages are necessary for virulence in multispecies infections. We show that a serine transporter, conserved among gammaproteobacteria, enables self-recognition. Molecular co-option of nutrient uptake could limit the sequence variation of these message proteins. SdaC is the primary transporter for l-serine, a vital metabolite for colonization during disease. Unlike many self-recognition receptors, SdaC is sufficiently conserved between species to achieve recognition. The predicted open conformation is shared by transport and recognition. SdaC reveals the interdependence of communication and nutrient acquisition. As the broader interactions of self-recognition proteins are studied, features shared among microbial self-recognition systems, such as those of spp. and spp., could emerge.
Topics: Bacterial Proteins; Biological Transport; Gene Expression Regulation, Bacterial; Locomotion; Membrane Proteins; Proteus mirabilis
PubMed: 34662238
DOI: 10.1128/JB.00347-21