A Case of Infective Endocarditis Caused by Citrobacter koseri: Unraveling a Rare Pathogen and Dire Outcome.

Citrobacter koseri (formerly classified as Citrobacter diversus) is a gram-negative bacillus (GNB) that occurs as an opportunistic pathogen in neonates and immunocompromised patients. Citrobacter species have been implicated in nosocomial settings leading to infections involving the urinary tract, respiratory tract, liver, biliary tract, meninges, and even in rarer conditions-blood stream infection and infective endocarditis (IE). Gram-negative bacilli are responsible for 3% to 4% of all IE cases and have been traditionally associated with intravenous drug users. Patients with non-HACEK (species other than Haemophilus species, Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens, or Kinglella species) GNB IE have poor clinical outcomes with higher rates of in-hospital mortality and complications. The American Heart Association (AHA) and Infectious Diseases Society of America (IDSA) both recommend the use of combination antibiotic therapy with a beta-lactam (penicillins, cephalosporins, or carbapenems) and either an aminoglycoside or fluoroquinolones for 6 weeks (about 1 and a half months) to treat IE due to non-HACEK GNB. Citrobacter koseri is becoming more recognized due to its inherent resistance to ampicillin and emerging drug resistance to beta lactams and aminoglycosides requiring carbapenem therapy. Our case is of a 75-year-old male with no previously reported history of primary or secondary immunodeficiency disorders who developed C koseri blood stream infection. His infectious work-up revealed mitral valve IE and septic cerebral emboli resulting in ischemic infarcts. This case illustrates the importance of recognizing GNB organisms as rising human pathogens in IE cases even without active injection drug use or nosocomial exposure.

Citrobacter koseri. Una endoftalmitis exógena inusual / Citrobacter koseri, an unusual exogenous endoftalmitis

Citrobacter koseri es un bacilo gramnegativo que causa endoftalmitis de forma muy infrecuente y agresiva, asociando mal pronóstico visual. Solo el 6% de las endoftalmitis son por gramnegativos y nuestro germen representa un pequeño porcentaje. Presentamos un varón de 65 años que trabaja en un laboratorio de animales. Acude a urgencias por pérdida de visión del ojo izquierdo debido a una hemorragia vítrea y desprendimiento de retina. Se practica una vitrectomía y 3 días después, desarrolla una endoftalmitis. Se realiza tratamiento intravítreo con inyecciones de vancomicina y ceftazidima así como tratamiento antibiótico tópico. Veinticuatro horas después, regresa con perforación corneal y una extrusión del cristalino. Ante la gravedad del cuadro se realiza una evisceración. El cultivo de las muestras confirman el microorganismo. Suponemos que la puerta de entrada del germen fueron las esclerotomías por el material de sutura expuesto, tras la manipulación de heces de roedores.(AU)

Citrobacter koseri is a bacillus that causes infrequent endophthalmitis. Six percent of cultures in endophthalmitis are Gram -, and as in these, Citrobacter koseri is associated with a poor visual prognosis. We present a 65-year-old man who works in an animal laboratory. He went to emergencies with loss of vision in his left eye due to a vitreous hemorrhage. A vitrectomy was performed and 3 days later, endophthalmitis was diagnosed. Vancomycin and ceftazidime were applied in eye drops and in two intravitreal injections. Twenty-four hours later he returned with a lens extrusion. Due to the severity of the condition, an evisceration was performed. Subsequently, the samples confirm the microorganism. We assume that the entry point for the bacterium was the sclerotomies through the exposed suture material, after handling rodent feces.(AU)

Citrobacter koseri, an unusual exogenous endoftalmitis.

Citrobacter koseri is a bacillus that causes infrequent endophthalmitis. 6% of cultures in endophthalmitis are Gram -, and as in these, C. koseri is associated with a poor visual prognosis. We present a 65-year-old man who works in an animal laboratory. He went to emergencies with loss of vision in his left eye due to a vitreous hemorrhage. A vitrectomy was performed and 3 days later, endophthalmitis was diagnosed. Vancomycin and Ceftazidime were applied in eye drops and in two intravitreal injections. 24 h later he returned with a lens extrusion. Due to the severity of the condition, an evisceration was performed. Subsequently, the samples confirm the microorganism. We assume that the entry point for the bacterium was the sclerotomies through the exposed suture material, after handling rodent feces.

Citrobacter koseri inhibits the growth of Staphylococcus epidermidis by suppressing iron utilization.

The human skin microbiome consists of many species of bacteria, including Staphylococcus aureus and S. epidermidis. Individuals with atopic dermatitis (AD) have an increased relative abundance of S. aureus, which exacerbates the inflammation of AD. Although S. epidermidis, a main component of healthy skin microbiota, inhibits the growth of S. aureus, the balance between S. epidermidis and S. aureus is disrupted in the skin of individuals with AD. In this study, we found that Citrobacter koseri isolated from patients with AD produces substances that inhibit the growth of S. epidermidis. Heat-treated culture supernatant (CS) of C. koseri inhibited the growth of S. epidermidis but not S. aureus. The genome of C. koseri has gene clusters related to siderophores and the heat-treated CS of C. koseri contained a high concentration of siderophores compared with the control medium. The inhibitory activity of C. koseri CS against the growth of S. epidermidis was decreased by the addition of iron, but not copper or zinc. Deferoxamine, an iron-chelating agent, also inhibited the growth of S. epidermidis, but not that of S. aureus. These findings suggest that C. koseri inhibits the growth of S. epidermidis by interfering with its iron utilization.

Citrobacter koseri related abortion and fetal septicemia in cattle.

A 31-month-old Holstein dairy cow aborted at 224 days of gestation with ejection of cheese-like lochia. Citrobacter koseri, which commonly exists in the normal flora of human and animal digestive tracts, was isolated from aborted fetal tissues (liver, spleen, kidney, heart, lung, cerebrum, and skeletal muscle) and fetal membranes. Histopathological examination revealed suppurative fibrinous meningoencephalitis of the cerebrum, cerebellum, and brainstem; suppurative bronchopneumonia; suppurative chorioamnionitis; and fibrous splenic serositis. Numerous gram-negative bacilli were detected in the cytoplasm of macrophages and/or neutrophils in these lesions. Bacteriological investigation and immunohistochemical staining identified the bacilli as C. koseri. This is the first report of cattle abortion caused by C. koseri infection in dairy cattle.

CkP1 bacteriophage, a S16-like myovirus that recognizes Citrobacter koseri lipopolysaccharide through its long tail fibers.

Citrobacter koseri is an emerging Gram-negative bacterial pathogen, which causes urinary tract infections. We isolated and characterized a novel S16-like myovirus CKP1 (vB_CkoM_CkP1), infecting C. koseri. CkP1 has a host range covering the whole C. koseri species, i.e., all strains that were tested, but does not infect other species. Its linear 168,463-bp genome contains 291 coding sequences, sharing sequence similarity with the Salmonella phage S16. Based on surface plasmon resonance and recombinant green florescence protein fusions, the tail fiber (gp267) was shown to decorate C. koseri cells, binding with a nanomolar affinity, without the need of accessory proteins. Both phage and the tail fiber specifically bind to bacterial cells by the lipopolysaccharide polymer. We further demonstrate that CkP1 is highly stable towards different environmental conditions of pH and temperatures and is able to control C. koseri cells in urine samples. Altogether, CkP1 features optimal in vitro characteristics to be used both as a control and detection agent towards drug-resistant C. koseri infections. KEY POINTS: • CkP1 infects all C. koseri strains tested • CkP1 recognizes C. koseri lipopolysaccharide through its long tail fiber • Both phage CkP1 and its tail fiber can be used to treat or detect C. koseri pathogens.

Genotypic Detection of qnrA and qnrC Genes in Citrobacter koseri Isolated from Patients with Urinary Tract Infection.

The urinary tract infection (UTI) is a prevalent infection that affects people of all ages. Bacterial agents are the most common causes of UTIs. Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, and other staphylococcal species, Citrobacter freundii and Citrobacter koseri (C. koseri) account for a smaller number of infections. These pathogens are transported into the urinary tract from the colonic biotope into dysbacteriosis. Urine samples were randomly collected from 249 outpatients who were suspected of having UTIs. After genital cleaning, 10 mL of urine specimens were collected in a sterilized bowel. Then, the specimens were centrifuged at 2,000 rpm for 5 min and the residue was aerobically incubated with the broth infusion of brain flasks at 37°C for 24 h and then applied with a sterile ring onto blood agar plates and MacConkey agar (OxoidTM). Out of 249 urine samples, the results proved that there were 176 (70.7%) and 51(20.5%) gram-negative and gram-positive bacteria isolates, respectively. However, the results demonstrated that there were 22 (8.8%) urine samples with no growth. In addition, the results showed that eight various antimicrobials are used to treat C. koseri. In the current study, C. koseri was treated with eight different antimicrobial agents. The antimicrobial resistance rate for 7 isolates against Cefotaxime, Ceftriaxone, Ciprofloxacin, and Levofloxacin was high for 6 (85.71%) isolates. The results indicated that 6 and 5 isolates had 85.71% and 71.42% antimicrobial resistance against Ceftazidime and Levofloxacin, respectively. Whereas Gentamicin showed a moderate rate of resistance (4 isolates, 57.14%), and Amikacin resistance was found in 5 isolates, accounting for 28.57%. The bacterial isolates had a high susceptibility rate to Imipenem. The qnrA gene was found in 6 (85.71%) isolates. However, the recorded data demonstrated that there is no isolate carrying the qnrC gene. Among all pathogenic bacteria, C. koseri was the lowest causative agent of UTI in this study and was highly resistant to most antimicrobials except Imipenem, which was a good antibiotic with 100% sensitivity.

Complete Genome Sequencing and Comparative Analysis of Citrobacter koseri CKNJ, a Strain Isolated from a Patient with Endogenous Endophthalmitis.

Citrobacter koseri (C. koseri) is an opportunistic pathogen that can cause a variety of diseases. Although the mortality rate of C. koseri infections is high, there is a paucity of clinical information. Furthermore, the genomic features of this species are poorly understood. Herein, we present a patient with endogenous endophthalmitis secondary to septicemia, and collected a C. koseri isolate, CKNJ, from the blood of the patient. Whole genome sequencing revealed that CKNJ harbors no plasmids and codes for 67 putative virulence factors. Whole genome single nucleotide polymorphism-based phylogenetic analysis revealed that the CKNJ strain was close to strains with the same isolation sites. Compared to the other sequenced C. koseri chromosomes, CKNJ contains several strain-variable regions, including one prophage and 2 large genomic islands. Sequencing of the first complete genome of a clinical strain from China should reinforce our understanding of the genomic features and pathogenicity of this invasive infection-causing C. koseri with clinical significance.

Citrobacter koseri Surgical Site Infection after Femoropopliteal Vein Bypass for Popliteal Artery Aneurysm Treated with a Graft Preserving Strategy.

Background: A 69-year-old man underwent ligation and evacuation of a popliteal artery aneurysm with a femoral-to-popliteal vein bypass. He had a history of Citrobacter koseri prostatitis two months prior to the surgery. One month postoperatively, he presented with extremity swelling, redness, and fluid collections around the graft. Methods: A graft preserving strategy was adopted. The patient underwent operative drainage, washing, and received long-term antibiotic therapy. Fluid culture grew Citrobacter koseri, previously not reported as cause of surgical site infection with infrainguinal graft involvement. Results: The infection was treated successfully, and the patient is remains symptom free 18 months post-operatively. Conclusions: This case highlights the importance of considering culturing the aneurysm content in the presence of infectious history.

In vitro antimicrobial susceptibility of clinical respiratory isolates to ceftazidime-avibactam and comparators (2016-2018).

BACKGROUND: This antimicrobial surveillance study reports in vitro antimicrobial activity and susceptibility data for a panel of agents against respiratory isolates of Enterobacterales and Pseudomonas aeruginosa. METHODS: Isolates from respiratory specimens were collected in Africa/Middle East, Asia/South Pacific, Europe and Latin America between 2016 and 2018, as part of the Antimicrobial Testing Leadership and Surveillance (ATLAS) program. Broth microdilution methodology was used to quantify minimum inhibitory concentrations, from which rates of susceptibility were determined using EUCAST breakpoints (version 10). Rates of subsets with genes encoding ß-lactamases (extended-spectrum ß-lactamases [ESBLs], serine carbapenemases and metallo-ß-lactamases [MBLs]) were also determined, as well as rates of multidrug-resistant (MDR) P. aeruginosa. RESULTS: Among all respiratory Enterobacterales isolates, susceptibility to ceftazidime-avibactam, meropenem, colistin and amikacin was ≥94.4% in each region. For Enterobacterales isolates that were ESBL-positive or carbapenemase-positive/MBL-negative, ceftazidime-avibactam susceptibility was 93.6 and 98.9%, respectively. Fewer than 42.7% of MBL-positive Enterobacterales isolates were susceptible to any agents, except colistin (89.0% susceptible). Tigecycline susceptibility was ≥90.0% among Citrobacter koseri and Escherichia coli isolates, including all ß-lactamase-positive subsets. ESBL-positive Enterobacterales were more commonly identified in each region than isolates that were ESBL/carbapenemase-positive; carbapenemase-positive/MBL-negative; or MBL-positive. Among all respiratory P. aeruginosa isolates, the combined susceptibility rates (susceptible at standard dosing regimen plus susceptible at increased exposure) were highest to ceftazidime-avibactam, colistin and amikacin (≥82.4% in each region). Susceptibility to colistin was ≥98.1% for all ß-lactamase-positive subsets of P. aeruginosa. The lowest rates of antimicrobial susceptibility were observed among MBL-positive isolates of P. aeruginosa (≤56.6%), with the exception of colistin (100% susceptible). MDR P. aeruginosa were most frequently identified in each region (18.7-28.7%), compared with the subsets of ESBL-positive; carbapenemase-positive/MBL-negative; or MBL-positive isolates. CONCLUSIONS: Rates of susceptibility among the collections of respiratory Enterobacterales and P. aeruginosa isolates were highest to ceftazidime-avibactam, colistin and amikacin in each region. Tigecycline was active against all subsets of C. koseri and E. coli, and colistin was active against all subsets of P. aeruginosa. The findings of this study indicate the need for continued antimicrobial surveillance among respiratory Gram-negative pathogens, in particular those with genes encoding MBLs.

Characterization of KPC-82, a KPC-2 Variant Conferring Resistance to Ceftazidime-Avibactam in a Carbapenem-Nonsusceptible Clinical Isolate of Citrobacter koseri.

KPC-82 is a KPC-2 variant identified in a carbapenem-nonsusceptible Citrobacter koseri that confers high-level resistance to ceftazidime-avibactam. Genomic analysis revealed that blaKPC-82 is carried by a chromosomally integrated Tn4401 transposon (disrupting porin gene phoE) and evolved by a 6-nucleotide tandem repeat duplication causing a two-amino-acid insertion (Ser-Asp) within the Ala267-Ser275 loop. Similar to related KPC variants, KPC-82 showed decreased carbapenemase activity when expressed in a heterologous background and remained susceptible to carbapenem/ß-lactamase inhibitor combinations.

Citrobacter koseri stimulates dendritic cells to induce IL-33 expression via abundant ATP production.

Introduction. Food allergies (FAs) occur due to intestinal immune dysfunction elicited by dysbiotic conditions. It was previously determined by us that Citrobacter species propagate in the faeces of mice with FAs and worsen allergic symptoms by inducing the allergenic cytokine IL-33. Dendritic cells can play important roles in regulation of FA responses.Hypothesis. Citrobacter species propagating in intestines of mice worsen allergic symptoms by stimulating dendritic cells to induce IL-33 expression.Aim. The aim of the present study was to analyse whether C. koseri stimulates dendritic cells to induce IL-33 expression.Methodology. IL-33 expression was evaluated in a DC2.4 mouse dendritic cell line stimulated by live or heat-inactivated C. koseri JCM1658, ATP, LPS extracted from C. koseri JCM1658 or other enterobacteria by real-time PCR. The ATP concentration and number of live bacteria in the culture supernatant were measured simultaneously.Results. Live C. koseri JCM1658 induced higher levels of IL-33 expression than other enterobacteria tested, but such a response was not elicited by heat-inactivated C. koseri JCM1658. LPS extracted from C. koseri JCM1658 did not induce IL-33 expression and suppressed live C. koseri JCM1658-induced IL-33 expression via the activation of Toll-like receptor 4 signalling. Furthermore, ATP produced by C. koseri JCM1658 stimulated dendritic cells to induce IL-33 expression by stimulating the P2X7 receptor, and LPS attenuated extracellular ATP-induced IL-33 expression. C. koseri JCM1658 was observed to proliferate more vigorously and produce more ATP than other enterobacteria.Conclusion. C. koseri acts as an allergenic bacterium through ATP production, stimulating dendritic cells to induce IL-33 expression, while LPS released from inactivated C. koseri JCM1658 attenuates this allergenicity.

Infant Case of Co-infection with SARS-CoV-2 and Citrobacter koseri Urinary Infection.

Since 2019 coronavirus disease (COVID-19) is highly contagious with a high mortality rate. France has taken strict infection control measures. According to the report by the Center for Disease Control and Prevention, children are less affected with COVID-19 and seem to have less severe disease than adults. We reported the first confirmed infant case of co-infection with SARS-CoV-2 and Citrobacter koseri urinary infection in 6-week-old child admitted on 25 March 2020 with mild symptoms in the Pediatric COVID Unit of Amiens University Hospital, France.

Positivity of Carbapenemase-producing Enterobacteriaceae in Patients Following Exposure within Long-term Care Facilities in Seoul, Korea.

BACKGROUND: Carbapenemase-producing Enterobacteriaceae (CPE) are emerging as a worldwide threat. Long-term care facilities (LTCFs) are considered a reservoir for CPE and play a central role in transmission to acute care hospitals. We investigated the CPE positivity in patients exposed to CPE in LTCFs. Furthermore, we analyzed the CPE positivity rates in the environment exposed to CPE. METHODS: We collected rectal swab specimens from patients residing in LTCFs who were exposed to CPE. Environmental sampling was performed by infection control practitioners from sites classified as patient private space, common space in the patient room, common space other than patient rooms, and nursing station. Each sample was cultured on a Chrom Klebsiella pneumoniae carbapenemase (KPC) agar for CPE screening. The positive isolates were subjected to a polymerase chain reaction to identify the presence of blaKPC, blaVIM, blaIMP, blaOXA-48, and blaNDM and determine CPE genotype. RESULTS: From 65 index cases, a total of 24 hospitals and 481 patients were enrolled; 414 patients who had resided in the same patient room as a patient with confirmed CPE and 67 patients who were newly admitted to that patient room. A total of 117 (24.3%) patients were positive for CPE among which 93 (22.5%, 93/414) were already admitted patients and 24 (35.8%, 24/67) were newly admitted patients. A total of 163 CPEs were detected and K. pneumoniae (n = 104, 63.8%) was the most common bacteria followed by Escherichia coli (n = 43, 26.4%) and Citrobacter koseri (n = 11, 6.7%). Environmental sampling was performed in 24 hospitals and 604 sites. A total of 12 sites (2.0%) were positive for CPE and sink in the nursing station (n = 6, 4.2%) was the most contaminated space. CONCLUSION: CPE colonization rates in patients exposed to CPE in LTCFs were higher than those found in acute care hospitals. Proper infection control measures for detecting and reducing CPE colonization in patients residing in LTCFs are required. Newly admitted patients could also be carriers; therefore, infection control for newly admitted patients also needs to be thorough.

Citrobacter diversus-derived keratinases and their potential application as detergent-compatible cloth-cleaning agents.

Currently, poultry farming is one of the sectors that have a significant impact on the global economy. In recent years, there has been an increase in the production of broilers, inflicting this segment of the industry to generate tons of keratin due to huge disposal of chicken feathers. This points to the need to degrade these chicken feathers, as they have emerged as a major threat to the environment. Thus, in this study we aimed to identify keratinases that are produced by the bacterium Citrobacter diversus and further investigate the biochemical characteristics of these keratin-degrading enzymes. In a submerged medium, the bacterium was capable of degrading chicken feathers almost completely after 36 h of fermentation. We found a maximum caseinolytic activity at pH 9-10.5 and 50-55 °C, and keratinolytic activity at pH 8.5-9.5 and 50 °C. Thus, given its stability at higher temperatures, upon incubation of this enzyme extract for 1 h at 50 °C, it showed approximately 50% of the keratinolytic and 100% of the caseinolytic activity. Further, under pH stability for 48 h at 4 °C, the enzyme extract maintained greater residual activity in the pH range 6-8. Caseinolytic activity was inhibited by EDTA and PMSF, whereas the keratinolytic activity was inhibited only by EDTA. Additionally, due to its alkaline activity and detergent compatibility, this enzyme extract could improve washing performance when added to a commercial detergent formulation. Using application tests, we could demonstrate a potential use of this bacterial enzyme extract as an additive in detergents to remove egg stains from cloth.

Covalent immobilization of recombinant Citrobacter koseri transaminase onto epoxy resins for consecutive asymmetric synthesis of L-phosphinothricin.

Transaminase responsible for alienating prochiral ketone compound is applicable to asymmetric synthesis of herbicide L-phosphinothricin (L-PPT). In this work, the covalent immobilization of recombinant transaminase from Citrobacter koseri (CkTA) was investigated on different epoxy resins. Using optimum ES-105 support, a higher immobilized activity was obtained via optimizing immobilization process in terms of enzyme loading, coupling time and initial PLP concentration. Crucially, due to blocking unreacted epoxy groups on support surface with amino acids, the reaction temperature of blocked immobilized biocatalyst was enhanced from 37 to 57 °C. Its thermostability at 57 °C was also found to be superior to that of free CkTA. The Km value was shifted from 36.75 mM of free CkTA to 39.87 mM of blocked immobilized biocatalyst, demonstrating that the affinity of enzyme to the substrate has not been apparently altered. Accordingly, the biocatalyst performed the consecutive synthesis of L-PPT for 11 cycles (yields>91%) with retaining more than 91.13% of the initial activity. The seemingly the highest reusability demonstrates this biocatalyst has prospective for reducing the costs of consecutive synthesis of L-PPT with high conversion.

Genome sequence of carbapenem-resistant Citrobacter koseri carrying blaOXA-181 isolated from sewage sludge.

OBJECTIVES: This study reported the resistome content of sewage sludge-isolated carbapenem-resistant Citrobacter koseri (C. koseri) carrying blaOXA-181. It also provided a general phylogenomic analysis highlighting antibiotic resistance genes (ARGs), plasmids and pathogenicity of C. koseri genomes. METHODS: The carbapenem-resistantC. koseri AS1 strain was isolated from sewage sludge on CHROMagar™ mSuperCARBA™ media. Whole genome sequencing of C. koseri AS1 was performed using an HiSeq X™ Ten instrument. Additional C. koseri genomes were downloaded from National Center for Biotechnology Information (NCBI). Phylogenomic analysis was established through CSI Phylogeny. ARGs, plasmids and pathogenicity were identified using ResFinder 3.1, PlasmidFinder 2.0 and PathogenFinder 1.1, respectively. RESULTS: The phylogenomic tree indicated a polyclonal pattern ofC. koseri genomes. Resistome analysis of C. koseri AS1 revealed ß-lactam resistance genes (blaMAL-1 and blaOXA-181) as well as a fosfomycin resistance gene (fosA7). Three plasmids (ColKP3, ColRNAI and IncX30) were identified in the C. koseri AS1 genome. In addition, 25 ARGs were found in downloaded genomes. Of these, clinically significant ARGs such as blaKPC-2 and blaOXA-48 were found in two and four genomes, respectively. Assessment of the genomes using PathogenFinder revealed all genomes as putative human pathogens. CONCLUSIONS: It is believed that noC. koseri genome has been reported to carry blaOXA-181; therefore, C. koseri AS1 is the first of its kind. This study also highlighted the resistome contents of C. koseri genomes.