Elizabethkingia Miricola Research Articles

Clinical characteristics of children with cystic fibrosis infected with unusual bacteria.

Pulmonary infections are usually caused by bacterial microorganisms such as Pseudomonas aeruginosa, Staphylococcus aureus, Haemophilus influenzae, and Burkholderia cepacia complex in cystic fibrosis (CF) patients. Unusual bacteria (UB) have been described by new isolation techniques recently in the respiratory samples of CF patients. The aim is to investigate the effects of the presence of UB in the respiratory cultures of CF patients on clinical outcomes, necessity of treatment and prognosis. The UB were identified by MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry technology. Rhizobium radiobacter were detected in 2, Chyrseobacterium species (gleum and indolgenes) in 5, Aeromonas hydrophila in 1, Orchobacterium anthropy in 1,Wautersiella falsenii in 1, Leclercia adecarboxylata in 1, Delftia acidovorans in 1, Cupriavidus Gilardi in 1, R.radiobacter twith Elizabethkingia miricola in 1 and R.radiobacter with C.gleum in 1 patient. Median age of the first UB growth was 3 years. After the first UB growth, the median follow-up time was 15 months. Before the UB growth, 60.0% of the patients had respiratory colonization with methicillin-susceptible S.aureus (MSSA). UB growth were accompanied with MSSA in 66.6% of the patients. Median percentage of FEV1 before and during the UB growth for patients who could perform spirometry, were 80 and 102, respectively. Median body mass index before and during the UB growth were 16 and 16.2, respectively. These UB were not detected during the follow-ups except in one patient. The UB growth did not cause any additional symptoms and decrease in BMI and FEV1 in patients with CF. MSSA may be a facilitating factor for UB growth as majority of the patients had MSSA colonization before and during the UB growth.

Susceptibility of Elizabethkingia spp. to commonly tested and novel antibiotics and concordance between broth microdilution and automated testing methods.

We aimed to determine susceptibilities of Elizabethkingia spp. to 25 commonly tested and 8 novel antibiotics, and to compare the performance of different susceptibility testing methods. Clinical isolates of Elizabethkingia spp., Chryseobacterium spp. and Flavobacterium spp. collected during 2002-18 (n = 210) in a nationwide surveillance programme in Taiwan were speciated by 16S rRNA sequencing. MICs were determined by broth microdilution. The broth microdilution results of 18 common antibiotics were compared with those obtained by the VITEK 2 automated system. Among the Elizabethkingia spp. identified (n = 108), Elizabethkingia anophelis was the most prevalent (n = 90), followed by Elizabethkingia meningoseptica (n = 7) and Elizabethkingia miricola cluster [E. miricola (n = 6), Elizabethkingia bruuniana (n = 3) and Elizabethkingia ursingii (n = 2)]. Most isolates were recovered from respiratory or blood specimens from hospitalized, elderly patients. PFGE showed two major and several minor E. anophelis clones. All isolates were resistant to nearly all the tested β-lactams. Doxycycline, minocycline and trimethoprim/sulfamethoxazole inhibited >90% of Elizabethkingia spp. Rifampin inhibited E. meningoseptica (100%) and E. anophelis (81.1%). Fluoroquinolones and tigecycline were active against E. meningoseptica and E. miricola cluster isolates. Novel antibiotics, including imipenem/relebactam, meropenem/vaborbactam, ceftazidime/avibactam, cefepime/zidebactam, delafloxacin, eravacycline and omadacycline were ineffective but lascufloxacin inhibited half of Elizabethkingia spp. The very major discrepancy rates of VITEK 2 were >1.5% for ciprofloxacin, moxifloxacin and vancomycin. Major discrepancy rates were >3% for amikacin, tigecycline, piperacillin/tazobactam and trimethoprim/sulfamethoxazole. MDR, absence of standard interpretation criteria and poor intermethod concordance necessitate working guidelines to facilitate future research of emerging Elizabethkingia spp.

Elizabethkingia miricola infection in multiple anuran species.

This report describes an outbreak of Elizabethkingia miricola in northern leopard frogs (Lithobates pipiens) and three other species of frogs and toads held in captivity in Germany. The authors examine several treatment options and underline the difficulties in treating larger numbers of individuals with antimicrobials applied through bathing. Whole genome sequencing of three bacterial isolates emphasizes their relatedness to other frog isolates and leads us to conclude that E. miricola is an emerging and difficult to treat pathogen with a broad host range across anuran species. Moreover, ambiguities in identification of flavobacteria associated with disease in frogs reported in the literature make it seem possible that E. miricola has been overlooked as an anuran pathogen in the past.

A real-time PCR assay for detection of emerging infectious Elizabethkingia miricola

Elizabethkingia miricola, a Gram-negative bacillus, is emerging as a life-threatening pathogen in both humans and animals. However, no specific and rapid diagnostic method exists to detect E. miricola. Here, we established a real-time PCR assay for the rapid, sensitive, and specific detection of E. miricola with a wide dynamic range of 108 copies/μL to 102 copies/μL. The detection limit of the real-time assay was 145 copies/μL, which was 100 times more sensitive than conventional PCR. All clinical isolates E. miricola from different host species yield very close Tm (80.25 ± 0.25 °C). Additionally, no cross-reaction or false positives were observed in the assay for non-target bacterial species. The performance of this assay was primarily assessed by testing frog tissue samples. Overall, our study provided a real-time PCR assay, which is a rapid, sensitive, and specific diagnostic method that could be used for early diagnosis and epidemiological investigation of E. miricola.

Bacteremia caused by Elizabethkingia miricola in a patient with acute pancreatitis and peritoneal dialysis

Bacteremia caused by Elizabethkingia miricola in a patient with acute pancreatitis and peritoneal dialysis

Whole-genome analysis of the potentially zoonotic Elizabethkingia miricola FL160902 with two new chromosomal MBL gene variants.

Elizabethkingia is an emerging life-threatening pathogen in both humans and animals. We describe the whole-genome analysis of an Elizabethkingia miricola strain isolated from a diseased frog in China and investigate the molecular mechanism of carbapenem resistance in this pathogen. WGS of E. miricola FL160902 was performed using single-molecule, real-time technology. A phylogenetic tree was generated by SNP analysis, comparing the genome of our strain with other E. miricola isolates of amphibian and human origins. Antimicrobial resistance genes and virulence-related genes were identified using the Comprehensive Antibiotic Resistance Database (CARD) and the Virulence Factor Database (VFDB). Two putative carbapenemase genes were expressed in Escherichia coli to evaluate their contribution to antimicrobial resistance. The genome of E. miricola FL160902 consists of a 4 249 586 bp circular chromosome with 27 putative resistance genes and 38 predicted virulence-associated genes. Comparative genomic analysis demonstrated that the E. miricola strains of human and amphibian origins have similar virulence-associated gene profiles. In addition, all the amphibian isolates clustered together with one of the human isolates in the phylogenetic analysis. WGS revealed the presence of two novel MBL genes, designated blaBlaB-16 and blaGOB-19. Cloning of blaBlaB-16 and blaGOB-19 into E. coli DH5α resulted in increased MICs of most β-lactams, including imipenem, meropenem and ampicillin. We identified two chromosomal MBL gene variants, named blaBlaB-16 and blaGOB-19 in an amphibian E. miricola isolate, which was considered potentially zoonotic based on phylogenetic analysis and virulence-associated gene comparison. This study highlights the importance of E. miricola as a potential zoonotic pathogen and a reservoir of MDR genes.

Elizabethkingia miricola BM10, a New Symbiotic Bacterium Isolated from the Hindgut of the Termite Reticulitermes speratus KMT001

Elizabethkingia miricola BM10, a symbiotic bacterium, has been isolated from the hindgut of Reticulitermes speratus KMT001, a termite which occurs on Bukhan Mountain in Seoul, Korea. This strain demonstrated a symbiotic characteristic, in that it lacked endo-β-1,4-glucanase activity, in a previous study. The major fatty acids of E. miricola BM10 were iso-C15:0, iso-C17:0 3-OH, and summed feature 3 (iso-C16:1ω7c/C16:1ω6c). The content of iso-C17:0 3-OH was higher, while those of ECL 13.566, iso-C17:11ω9c, and summed feature 4 were lower than the other three type-strains of the Elizabethkingia genus. The 16S rRNA phylogenetic analysis confirmed that E. miricola BM10 is a new species. The whole genome of E. miricola BM10 was sequenced. The average nucleotide identity of strain BM10 as evaluated by pairwise comparison with E. anophelis R26, E. meningoseptica ATCC 13253, and E. miricola GTC 862 was shown to be 91.5%, 81.2%, and 94.29%, respectively. Based on our study results, E. miricola BM10 appears to represent a new strain of the genus Elizabethkingia.

Uncertainties in Identification of Elizabethkingia meningoseptica

To the Editors: With reference to the article entitled “Isolation of Methicillin Resistant Staphylococcus aureus and Multidrug Resistant Elizabethkingia meningoseptica From Neonates Within Minutes of Birth” by Kwambana-Adams et al,1 we would like to comment on the automated API (Analytical Profile Index) system (BioMerieux, Basingstoke, United Kingdom), used by the authors for identification of bacterial isolates including E. meningoseptica. According to a recent study by Lin et al,2 where they compared 4 automated systems with 16S rRNA gene sequencing for identification of Elizabethkingia species, overall correct rate for identification was 24.5% for API/ID 32 v3.1system. Although API system identified 12 of 13 E. meningoseptica isolates, all other Elizabethkingia species such as Elizabethkingia anophelis and Elizabethkingia miricola were also misidentified as E. meningoseptica because of absence of database coverage for these species. Moreover, an isolate of Chryseobacterium species was also misidentified as E. meningoseptica by the automated system. These data add to the identification inaccuracies of the API system with regard to Elizabethkingia species. To the best of our efforts, we could not find any other study which used API system for the identification of this organism. Even the review article3 cited by Kwambana-Adams et al1 did not mention the identification methods for Elizabethkingia species. Correct identification of the organism is crucial for the diagnosis and management strategies as E. anophelis has also recently emerged as a nosocomial neonatal pathogen.4 Hence, differentiation between E. anophelis and E. meningoseptica requires accurate microbial identification. MALDI-TOF MS (Matrix Assisted Laser Desorption/Ionization-Time Of Flight Mass Spectrometry) is a valuable tool for diagnosis of such unusual pathogens. However, more comparative studies are required with expanded databases of MALDI-TOF MS with species-specific spectra for reliable diagnosis of this pathogen. Also, we recommend database updation of API system for Elizabethkingia species other than E. meningoseptica. Gurmeet R. Rahim, PhDNeha Gupta, MDDepartment of MicrobiologyShah SatnamJi Speciality HospitalsSirsa, Haryana, India[email protected]

Elizabethkingia miricola infection in Chinese spiny frog (Quasipaa spinosa).

Elizabethkingia miricola is a Gram-negative rod which has been incriminated in severe infections in humans. Recently, a serious infectious disease was identified in Chinese spiny frogs (Quasipaa spinosa), in the Sichuan Province of China; the disease was characterized by corneal opacity, the presence of ascites and neurological symptoms. A Gram-negative bacillus was isolated from the liver, spleen and kidney of the diseased frogs. Experimental infection test revealed that the bacillus could infect the frogs Q.spinosa and the LD50 value was 1.19×106 cfu per frog. The isolated Gram-negative bacillus was identified as E.miricola according to phenotypic characteristics, 16S rRNA and gyrB gene sequence analysis. The isolated strain was only susceptible to florfenicol among all investigated chemotherapeutic agents. Histological examination revealed that E.miricola infection caused pathological lesions to multiple organs and tissues, especially in the liver, brain, kidney. These results confirmed that E.miricola is an emerging pathogen of Chinese spiny frogs.

Case Report: First report of Elizabethkingia miricola infection in a patient with cystic fibrosis

Elizabethkingia miricola is a rare non-fermenting Gram-negative rod that has previously been reported to be associated with blood stream and pulmonary abscess infections, but never before in cystic fibrosis (CF). Here we present the first reported case of Elizabethkingia miricola infection in a patient with CF and discuss the management options. We describe a patient with CF in whom we observed clinical and spirometric evidence of pulmonary exacerbation with the associated growth of E. miricola in sputum culture. The period of clinical instability was observed to coincide with the obtainment of four sputum samples from which E. miricola was cultured; improvement was seen following treatment with ciprofloxacin and the subsequent eradication of E. miricola. We conclude that E. miricola is able to survive in the CF lung and in this case was associated with pulmonary exacerbation. Empirical treatment with fluoroquinolones is appropriate, based on our experience.

Case Report: First report of Elizabethkingia miricola infection in a patient with cystic fibrosis.

Elizabethkingia miricola is a rare non-fermenting Gram-negative rod that has previously been reported to be associated with blood stream and pulmonary abscess infections, but never before in cystic fibrosis (CF). Here we present the first reported case of Elizabethkingia miricola infection in a patient with CF and discuss the management options. We describe a patient with CF in whom we observed clinical and spirometric evidence of pulmonary exacerbation with the associated growth of E. miricola in sputum culture. The period of clinical instability was observed to coincide with the obtainment of four sputum samples from which E. miricola was cultured; improvement was seen following treatment with ciprofloxacin and the subsequent eradication of E. miricola. We conclude that E. miricola is able to survive in the CF lung and in this case was associated with pulmonary exacerbation. Empirical treatment with fluoroquinolones is appropriate, based on our experience.

Elizabethkingia miricola as an opportunistic oral pathogen associated with superinfectious complications in humoral immunodeficiency: a case report

BackgroundElizabethkingia miricola is a rare Gram-negative bacterium found in water and clinical specimens. Typical culturing methods often misidentify Elizabethkingia spp. as Flavobacterium or Chryseobacterium. Although diagnosis is based on culturing samples taken from sterile sites, such as blood, a proper identification of this bacterium requires an expertise that goes beyond the capabilities of a typical clinical laboratory.Case presentationA 35-year-old woman diagnosed with common variable immunodeficiency was admitted to our center. Previous treatment with antibiotics (amoxicillin plus clavulanate, first and third generation of cephalosporins, macrolides) and systemic corticosteroids (up to 120 mg/day of prednisolone) failed to arrest the spread of inflammation. Gingival recession was observed in her oral cavity, resulting in an apparent lengthening of her teeth. In addition to typical commensal bacteria, including streptococci and neisseriae, strains of Rothia mucilaginosa and Elizabethkingia miricola were identified upon a detailed microbiological examination using a MALDI-TOF MS Biotyper system. The presence of the latter strain correlated with severe periodontitis, lack of IgA in her saliva and serum, a very low IgG concentration (< 50 mg/dl), IgM-paraproteinemia, decreases in C3a and C5a and microvascular abnormality. High-dose immunoglobulin (to maintain IgG > 500 mg/dl) and targeted levofloxacin treatment resulted in immune system reconstitution, oral healing, and eradication of the Elizabethkingia infection.ConclusionsE. miricola rarely causes disease in healthy individuals. However, the overgrowth of commensal bacteria, lack of IgG/IgA, microvasculopathy and complement cascade activation in patients with humoral immunodeficiency may facilitate Elizabethkingia invasion. Overuse of antibiotics, particularly beta-lactams, may cause mucosal colonization by E. miricola, followed by its multiplication combined with periodontitis that prompts bacterial translocation. MALDI-TOF Biotyper analysis may become a method of choice for identification of Elizabethkingia infections.

Pathogenic Elizabethkingia miricola Infection in Cultured Black-Spotted Frogs, China, 2016

Multiregional outbreaks of meningitis-like disease caused by Elizabethkingia miricola were confirmed in black-spotted frog farms in China in 2016. Whole-genome sequencing revealed that this amphibian E. miricola strain is closely related to human clinical isolates. Our findings indicate that E. miricola can be epizootic and may pose a threat to humans.

Revisiting the taxonomy of the genus Elizabethkingia using whole-genome sequencing, optical mapping, and MALDI-TOF, along with proposal of three novel Elizabethkingia species: Elizabethkingia bruuniana sp. nov., Elizabethkingia ursingii sp. nov., and Elizabethkingia occulta sp. nov.

The genus Elizabethkingia is genetically heterogeneous, and the phenotypic similarities between recognized species pose challenges in correct identification of clinically derived isolates. In addition to the type species Elizabethkingia meningoseptica, and more recently proposed Elizabethkingia miricola, Elizabethkingia anophelis and Elizabethkingia endophytica, four genomospecies have long been recognized. By comparing historic DNA-DNA hybridization results with whole genome sequences, optical maps, and MALDI-TOF mass spectra on a large and diverse set of strains, we propose a comprehensive taxonomic revision of this genus. Genomospecies 1 and 2 contain the type strains E. anophelis and E. miricola, respectively. Genomospecies 3 and 4 are herein proposed as novel species named as Elizabethkingia bruuniana sp. nov. (type strain, G0146T=DSM2975T=CCUG69503T=CIP 111191T) and Elizabethkingia ursingii sp. nov. (type strain, G4122T=DSM2974T=CCUG69496T=CIP111192T), respectively. Finally, the new species Elizabethkingia occulta sp. nov. (type strain G4070T=DSM2976T=CCUG69505T=CIP111193T), is proposed.

Decolourisations and biodegradations of model azo dye solutions using a sequence batch reactor, followed by ultrafiltration

The main objective of this study was to investigate the efficiency of biological treatment of azo dye-containing wastewater with a sequencing batch reactor system, followed by ultrafiltration. The performance of the system was quantified by measuring the chemical oxygen demand and azo dye concentration. The biodegradation was carried out under combined alternating anaerobic and aerobic conditions with Nylosan Yellow E2RL SGR as a model azo dye contaminant. The bioprocess revealed a maximal reduction in chemical oxygen demand and dye removal efficiency of 91 and 85%, respectively. After ultrafiltration of effluent from the biological treatment, the efficiency increased to 94% for chemical oxygen demand and to 97% for the azo dye decolourisation. Samples of activated sludge from the bioprocess were collected for microbial characterisation. Bacteria and fungi were isolated and identified by 16S rRNA gene and ITS1-5.8S rDNA-ITS2 sequence analysis, respectively. Serratia marcescens and Klebsiella oxytoca were the most common bacteria with the highest number present during the aerobic and anaerobic phases of the bioprocess. In addition, a high number of Elizabethkingia miricola, Morganella morganii, Comamonas testosteroni, Trichosporon sp. and Galactomyces sp. were detected. Taken together, our results demonstrated that the sequencing batch reactor system combined with ultrafiltration is an efficient technique for treatment of wastewater containing azo dye. Moreover, the ultrafiltration effectively removes the microbiota from the final effluent resulting in stable product water.

Determination of Elizabethkingia Diversity by MALDI-TOF Mass Spectrometry and Whole-Genome Sequencing.

In a hospital-acquired infection with multidrug-resistant Elizabethkingia, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rRNA gene analysis identified the pathogen as Elizabethkingia miricola. Whole-genome sequencing, genus-level core genome analysis, and in silico DNA-DNA hybridization of 35 Elizabethkingia strains indicated that the species taxonomy should be further explored.

Genome of the carbapenemase-producing clinical isolate Elizabethkingia miricola EM_CHUV and comparative genomics with Elizabethkingia meningoseptica and Elizabethkingia anophelis: evidence for intrinsic multidrug resistance trait of emerging pathogens

Elizabethkingia miricola is a Gram-negative non-fermenting rod emerging as a life-threatening human pathogen. The multidrug-resistant (MDR) carbapenemase-producing clinical isolate E. miricola EM_CHUV was recovered in the setting of severe nosocomial pneumonia. In this study, the genome of E. miricola EM_CHUV was sequenced and a functional analysis was performed, including a comparative genomic study with Elizabethkingia meningoseptica and Elizabethkingia anophelis. The resistome of EM_CHUV revealed the presence of a high number of resistance genes, including the presence of the blaGOB-13 and blaB-9 carbapenemase-encoding genes. Twelve mobility genes, with only two of them located in the proximity of resistance genes, and four potential genomic islands were identified in the genome of EM_CHUV, but no prophages or CRISPR sequences. Ten restriction–modification system (RMS) genes were also identified. In addition, we report the presence of a putative conjugative plasmid (pEM_CHUV) that does not encode any antibiotic resistance genes. Altogether, these findings point towards a limited number of DNA exchanges with other bacteria and suggest that multidrug resistance is an intrinsic trait of E. miricola owing to the presence of a high number of resistance genes within the bacterial core genome.

Relative Prevalence and Antimicrobial Susceptibility of Clinical Isolates of Elizabethkingia Species Based on 16S rRNA Gene Sequencing.

Some of the previously reported clinical isolates of Elizabethkingia meningoseptica may be later named species of Elizabethkingia We determined the accuracy of species identification (with two matrix-assisted laser desorption ionization-time of flight mass spectrometry [MALDI-TOF MS] systems and the Vitek 2 GN card), relative prevalence of three Elizabethkingia spp. in clinical specimens, and antimicrobial susceptibility of the species identified by 16S rRNA gene sequencing. Specimens for culture were collected from patients in a university hospital in Seoul, South Korea, between 2009 and 2015. All 3 Elizabethkingia spp. were detected in patients; among the 86 isolates identified by 16S rRNA gene sequencing, 17 (19.8%) were E. meningoseptica, 18 (20.9%) were Elizabethkingia miricola, and 51 (59.3%) were Elizabethkingia anophelis Only the MALDI-TOF Vitek MS system with an amended database correctly identified all of the isolates. The majority (76.7%) of the isolates were from the lower respiratory tract, and 8 (9.3%) were from blood. Over 90% of E. meningoseptica and E. anophelis isolates were susceptible to piperacillin-tazobactam and rifampin. In contrast, all E. miricola isolates were susceptible to fluoroquinolones except ciprofloxacin. Further studies are urgently needed to determine the optimal antimicrobial agents for the treatment of infections due to each individual Elizabethkingia species.

Autofluorescence Signatures of Seven Pathogens: Preliminary in Vitro Investigations of a Potential Diagnostic for Acanthamoeba Keratitis.

Acanthamoeba keratitis can cause devastating damage to the human cornea and is often difficult to diagnose by routine clinical methods. In this preliminary study, we investigated whether Acanthamoeba may be distinguished from other common corneal pathogens through its autofluorescence response. Although only a small number of pathogens were studied, the identification of a unique Acanthamoeba signature would indicate that autofluorescence spectroscopy as a diagnostic method merits further investigation. Samples of 7 common pathogens (Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Elizabethkingia miricola, Achromobacter ruhlandii, Candida albicans, and Acanthamoeba castellanii) in solution were excited with ultraviolet light at a number of successive, narrow wavebands between 260 and 400 nm, and their fluorescence response recorded. Principal Component Analysis was used to allow better visualization of the differences in response to UV light for different species. Acanthamoeba was found to possess a characteristic autofluorescence response and was easily distinguished from E. coli, S. aureus, P. aeruginosa, E. miricola, A. ruhlandii, and C. albicans over a wide range of excitation wavelengths. We also found a clear discrimination between E. coli, C. albicans, and P. aeruginosa at an excitation wavelength of 274 nm, whereas E. miricola, S. aureus, and A. ruhlandii could be separated using an excitation wavelength of 308 nm. Our results, although preliminary, indicate that autofluorescence spectroscopy shows promise as a diagnostic technique for keratitis. We intend to expand the set of pathogens studied before assessing the feasibility of the technique in vivo by introducing cultures onto pig corneas.

BlaB-15, a new BlaB metallo-β-lactamase variant found in an Elizabethkingia miricola clinical isolate

A multidrug-resistant strain of Elizabethkingia miricola was isolated from the urine of a 2-year-old boy hospitalized for severe clinical conditions. The strain produces 2 metallo-β-lactamases belonging to subclasses B1 and B3: a new BlaB variant (BlaB-15) and a GOB-7–like enzyme.