A Core Genome Multilocus Sequence Typing Scheme for Proteus mirabilis

Objective A core genome multilocus sequence typing(cgMLST)scheme to genotype and identify potential risk clonal groups(CGs)in Proteus mirabilis.Methods In this work,we propose a publicly available cgMLST scheme for P.mirabilis using chew BBACA.In total 72 complete P.mirabilis genomes,representing the diversity of this species,were used to set up a cgMLST scheme targeting 1,842 genes,635 unfinished(contig,chromosome,and scaffold)genomes were used for its validation.Results We identified a total of 205 CGs from 695 P.mirabilis strains with regional distribution characteristics.Of these,159 unique CGs were distributed in 16 countries.CG20 and CG3 carried large numbers of shared and unique antibiotic resistance genes.Nine virulence genes(papC,papD,papE,papF,papG,papH,papI,papJ,and papK)related to the P fimbrial operon that cause severe urinary tract infections were only found in CG20.These CGs require attention due to potential risks.Conclusion This research innovatively performs high-resolution molecular typing of P.mirabilis using whole-genome sequencing technology combined with a bioinformatics pipeline(chewBBACA).We found that the CGs of P.mirabilis showed regional distribution differences.We expect that our research will contribute to the establishment of cgMLST for P.mirabilis.

Phenotypic and Antimicrobial Susceptibility Studies of Proteus mirabilis Isolates from Fresh Water Fishes in FCT, Abuja-Nigeria

Our study was carried out to determine the phenotypic characterization antimicrobial susceptibility of Proteus mirabilis from fish in FCT, Abuja using isolation, selective plating, preliminary observation, complete biochemical method and antimicrobial susceptibility testing. The biochemical tests conducted includes include Citrate Utilization test, Triple Sugar Iron test, Urea test, Methyl Red test, Indole test, and Voges Proskauer test. The isolates were confirmed by MicrobactTMGNB24E identification kit (Oxiod, UK). A total of 400 fish samples were bought in the market from three area council of the FCT. The result of the study showed overall prevalence rate of (13) 3.25% of Proteus mirabilis isolates. Distribution based on Area councils showed that AMAC had higher prevalence rate of 4.81%, while Bwari had 2.99% and Gwagwalada with 2.57% prevalence. All isolates were subjected to antimicrobial susceptibility testing using the modified single disc diffusion method. From the antimicrobial susceptibility testing done it was discovered that Proteus mirabilis are resistant to Amoxyclav (100%), Erythromycin (92.3%), Tetracycline (92.3%) and Ceftriaxone (23.1%). However, the isolates were susceptible to Ofloxacin (100%), Netillin (92.3%), Levofloxacin (92.3%), Ceftazidime (76.9%), Co-trimoxazole (69.2%) and Gentamicin (61.5%). Since Proteus mirabilis sources of zoonotic diseases and can potentially be dangerous to humans and other animals, our research was able to isolate it from fresh water fish sold in the Federal Capital Territory. This makes public health awareness of the risks associated with Proteus mirabilis in Nigeria necessary.

Identification of the autotransporter Pet toxin in Proteus mirabilis strain isolated from patients with urinary tract infections

Proteus mirabilis, a motile Gram-negative bacterium, represents a common cause of complicated urinary tract infections. Autotransporters are a family of secreted proteins from Gram-negative bacteria that direct their own secretion across the outer membrane (type V autotransporter secretion mechanism). Serine protease autotransporters of Enterobacteriaceae (SPATEs) include adhesins, toxins, and proteases that can contribute to the virulence. Plasmid-encoded toxin (Pet) is the predominant protein in culture supernatants of enteroaggregative E. coli prototype strain 042 and has been extensively studied. Pet toxin is encoded on the 65-MDa adherence-related plasmid of EAEC 042 strain. In this work, Pet protein was found in the supernatant obtained from Proteus mirabilis RTX339 strain isolated from a psychiatric patient suffering complicated urinary tract infections (UTIs). The nucleotide sequence of pet gene was obtained using primers designed from E. coli 042 pet gene reported. The alignment of the sequence showed 100% identity with the pet gene reported. Is important to note that Proteus mirabilis RTX339 pet gene has chromosomal location. The chromosomal location of the gene was established since no plasmids were harbored by this strain.

Pathogenic infection and microbial composition of yellow catfish(Pelteobagrus fulvidraco)challenged by Aeromonas veronii and Proteus mirabilis

The catfishes are a group of economically important freshwater fish in China,which in recent years have suffered heavy losses as a result of bacterial outbreaks.In this study,we examined the diversity of the microbiome of infected skin mucus of yellow catfish(Pelteobagrus fulvidraco)and channel catfish(Ictalurus punctatus)and analyzed the bacterial pathogens.We found several common pathogenic bacteria,such as Aeromonas spp.,Vibrio spp.,Moraxella spp.and Proteus spp.present in both fish species,but with significantly different bacterial community structures.We isolated and cultured Aeromonas veronii and Proteus mirabilis and validated their infectivity in zebrafish(Danio rerio)and yellow catfish.Intraperitoneal injection of either bacteria into zebrafish,but not immersion,caused 100%mortality and ovary fragmentation.Yellow catfish was more sensitive than zebrafish with 100%mortality in the immersion challenges with A.veronii or P.mirabilis,and with a higher abundance of A.veronii in the P.mirabilis-challenged group compared to control.To our knowledge,this is the first study on the pathogenicity of P.mirabilis in yellow catfish,and the results will help to develop effective strategies for the disease prevention and control in catfish farming.

Emergence of Klebsiella pneumoniae carbapenemase-producing Proteus mirabilis in Hangzhou, China

Background Carbapenems are used to treat severe infections caused by multi-drug-resistant organisms, however, the emergence of carbapenem-resistant bacterial isolates is becoming an increasing therapeutic challenge. Since the first Klebsiella （K.） pneumoniae carbapenemase （KPC）-producing K. pneumoniae was reported in 2001, KPC-producing isolates have been found increasingly, specially in Enterobacteriaceae. The aim of this study was to characterize the mechanisms of a carbapenem-resistant Proteus （P.） mirabilis. Methods A carbapenem-resistant P. mirabilis isolate was recovered from pleural drainage fluid of a patient admitted to surgical intensive care unit. Antimicrobial susceptibility testing of the isolate was performed by disk diffusion according to Clinical and Laboratory Standards Institute guidelines, and subsequent minimal inhibitory concentrations were determined with the E-test. Amplification of the blaKPC gene generated a positive band and the PCR products were sequenced subsequently. The plasmid of the isolate was extracted and was successfully transformed into Escherichia （E.） coli DH5a. Results The P. mirabilis isolate was resistant to all detected antimicrobial agents except tigecycline. KPC-2 was confirmed by DNA sequence analysis. The transformant E. coil was resistant to carbapenems. Further study demonstrated that upstream and downstream regions of b/aKPC-2 were identical to that observed in K. pneumoniae submitted to GenBank from China in 2007. Conclusion Carbapenem resistance in the P. mirabilis isolate in this study is mainly due to production of KPC-2.

Multidrug-resistant Proteus mirabilis isolates carrying blaOXA-1 and blaNDM-1 from wildlife in China:increasing public health risk

The emergence of multidrug resistance(MDR)in Proteus mirabilis clinical isolates is a growing public health concern and has serious implications for wildlife.What is the role of wildlife has been become one of the hot issues in disseminating antimicrobial resistance.Here,54 P.mirabilis isolates from 12 different species were identified.Among them,25 isolates were determined to be MDR by profile of antimicrobial susceptibility;10 MDR P.mirabilis isolates were subjected to comparative genomic analysis by whole genome sequencing.Comprehensive analysis showed that chromosome of P.mirabilis isolates mainly carries multidrug-resistance complex elements harboring resistance to carbapenem genes blaOXA-1,blaNDM-1,and blaTEM-1.Class I integron is the insertion hotspot of IS26;it can be inserted into type I integron at different sites,thus forming a variety of multiple drug resistance decision sites.At the same time,Tn21,Tn7,and SXT/R391 mobile elements cause widespread spread of these drug resistance genes.In conclusion,P.mirabilis isolates from wildlife showed higher resistance to commonly used clinic drugs comparing to those from human.Therefore,wild animals carrying MDR clinical isolates should be paid attention to by the public health.

Comparative outcomes of the pathogen in cultured Jones tubes used in lacrimal bypass surgery according to follow up periods

AIM:To evaluate the pathogens in cultured Jones tubes used in lacrimal bypass surgery according to the postoperative periods and to obtain data for the prevention of infection of functional lacrimal stent invention.METHODS:Totally 71 patients(81 eyes)who underwent the removal of Jones tubes were enrolled in study.All the removed Jones tubes were cultured for bacterial and fungal identification and tested for bacterial antibiotic sensitivity.The results were analyzed according to the duration of the inserted Jones tube after lacrimal bypass surgery.RESULTS:Of the 81 eyes,bacteria were isolated from 69 eyes(85.2%)and fungi from 6 eyes(7.4%).Among 69 eyes,40.6% showed Staphylococcus aureus(S.aureus),11.6% were Pseudomonas aeruginosa(P.aeruginosa).Gram-positive bacteria were isolated more than Gramnegative bacteria,but Gram-negative bacteria showed a higher incidence in the Jones tube implanted for over 10y(P=0.035).The antibiotic sensitivity test showed that 46.4% of S.aureus were resistant to oxacillin.In terms of antibiotics commonly used in ocular clinical practice,vancomycin was sensitive to S.aureus and Streptococcus pneumoniae(S.pneumoniae),amikacin responded to P.aeruginosa and Proteus mirabilis(P.mirabilis).Trimethoprim/sulfamethoxazole(TMP/SMX)was all sensitive to S.aureus,S.pneumoniae and P.mirabilis except P.aeruginosa.CONCLUSION:S.aureus is the most commonly found organism in the Jones tube after lacrimal bypass surgery,and 46.4% of them are methicillin-resistant S.aureus(MRSA),sensitive to vancomycin.Especially,P.mirabilis responded with amikacin is dominantly detected in the Jones tubes implanted for more than 10y.

Pathological effects of vanillic acid on urinary tract in rats with infectious calculi

Objective:The pathological changes of urinary tract were observed after the treatment of infectious stone by vanillic acid.Methods:40 male Wistar rats were randomly divided into blank control group,infectious stone group,vanillic acid group and acetohydroxamic acid group.Blank control group:4 male Wistar rats were fed without special treatment.Infectious Stone Group:12 male Wistar rats were implanted with 4 mm bacteria bearing foreign body in the front of the polyurethane vein indwelling needle infected with Proteus mirabilis into the bladder to induce the formation of infection stone.In the vanillic acid group,after the model were establishmented,12 male Wistar rats were treated with vanillic acid at 50mg/kg?d for 3 weeks.Acetohydroxamic acid group:after the model were establishmented,12 male Wistar rats were treated with acetohydroxamic acid at 12.5mg/kg?d for 3 weeks.The rats were killed 21 days later.The kidney,ureter and bladder were fixed in 10%paraformaldehyde solution for 24 hours.The specimens were embedded in paraffin,and the pathological analysis was carried out after HE and immunohistochemical staining.Results:In addition to the blank control group,the other three groups of rats had calculus formation in the bladder.No obvious pathological changes were found in the kidney,ureter and bladder in the blank control group,and inflammatory reactions were found in the other three groups:severe urinary tract inflammation was found in the infectious stone group,followed by acetylhydroxamic acid group.In the vanillic acid group,the pathological changes of urinary tract were the lightest,with a small amount of inflammatory cell infiltration and mucosal hyperplasia,but no serious pathological changes such as mucosal loss.The expression of Tamm-Horsfall Protein(THP)in the ascending branch of loop of Henle and the proximal part of distal tubule in vanillic acid group was not significant compared with that in the infectious stone group and acetohydroxamic acid group.Conclusion:Proteus mirabilis and its infective stones can lead to the damage of urinary epithelium.Vanillic acid can reduce the inflammatory lesions of urinary tissue.

Stereoselective Synthesis and Immunological Evaluation of Common O-antigen of P. mirabilis OE and P. vulgaris TG103

Proteus species especially Proteus mirabilis and Proteus vulgaris are zoonotic pathogens which can cause public health disease.Owing to their antibiotic-resistance,developing vaccines against these pathogens is urgently required.Herein,we describe the first synthesis of the common O-antigen of Proteus mirabilis OE and Proteus vulgaris TG 103.

β-Cyclodextrin-modified AuBi metallic aerogels enable efficient peroxidase mimicking for colorimetric sensing of urease-positive pathogenic bacteria

The detection of pathogenic bacteria with improved accessibility,reduced analysis time,and increased sensitivity is of great importance for diagnosing the infected disease.Nanozymes have attracted rising attention in the bioassay field.Designing a model nanozyme needs the combined merit of sensible nanostructures and a large specific surface area to guarantee exceptional enzyme-mimic activity.Herein,aβ-cyclodextrin modified AuBi aerogel is prepared by a one-pot reduction strategy.The introduction ofβ-cyclodextrin(featured with a hydrophobic cavity and hydrophilic surface)enhances the catalytic activity of AuBi aerogels by engendering host-guest complex and improving dispersity/stability.Based on the specific urea hydrolysis,which could produce NH_(3)to raise pH by urease,the pH up-regulation would inhibit the peroxidase-mimicking performances ofβ-cyclodextrin/AuBi aerogels.Therefore,the sensitive colorimetric detection platform for urease activity could be constructed.Moreover,the sensing platform can detect straightforwardly urease-positive Proteus mirabilis in urine circumstances with a wide detection range and a low limit of detection(LOD)of 4 colony-forming unit(CFU)·mL^(-1).The reproducibility,stability,and specificity of this approach are verified to be satisfactory.Also,as an inhibitor of urease activity,the fluoride ion could be detected by the constructed sensing platform sensitively and specifically.Overall,this work provides a blueprint for designing an ideal nanozyme and paves a new roadway for detecting pathogenic bacteria.