Non-diphtheria Corynebacteria Research Articles

First case report of Corynebacterium minutissimum septic arthritis associated with an indwelling device.

Corynebacterium minutissimum usually presents with erythrasma, a macular rash, but rarely it can cause extracutaneous infections such as endocarditis, peritonitis, pyelonephritis, and catheter-related bloodstream infections. Here, we present the first case of septic arthritis caused by C. minutissimum and a literature review of invasive C. minutissimum infections. The diagnosis was confirmed by the patient's clinical findings and microbiological examination of the joint fluid. Considering the patients hemodialysis catheter and the pathogenic nature of the C. minutissimum, we strongly suspected a catheter-related infection. Clinical and laboratory follow-ups showed consistent improvement with treatment, characterized by reduced pain, swelling, and localized warmth in the left knee, along with a significant decline in acute-phase reactants. The patient achieved full recovery following a four-week course of antibiotics and catheter removal. Our report emphasizes the importance of not disregarding the isolation of non-diphtheria Corynebacterium from appropriately collected sterile samples in symptomatic patients, particularly those with indwelling devices. Based on this case report, clinicians should consider the rare possibility of this microorganism being the cause of septic arthritis.

Identification and Antibiogram of Clinically Relevant Non-Diphtheriae Corynebacterium species

Abstract Background and Objectives With the increase in immunosuppressed patients and antimicrobial misuse, Non-diphtheriae Corynebacterium species (NDC spp.) have risen as opportunistic pathogens in hospitalized patients. The present study aimed at comparing Vitek 2C automated identification cards versus MALDI-TOF as the gold standard method for the identification of different NDC spp. and determining the antimicrobial susceptibility of NDC spp. by the disc diffusion method for proper management of patients. Materials and Methods The study included 30 NDC spp. isolates recovered from different samples and were subjected to identification by MALDI-TOF and Vitek 2C. The disc diffusion method was performed to determine the antimicrobial susceptibility of the following antibiotics: pencillin G, tetracycline, gentamicin, ciprofloxacin, vancomycin, and Linezolid. Results Our study showed that Corynebacterium striatum (C. striatum) was the most commonly isolated species (83.3%), followed by C. amycolatum (10%). 36.7% of the isolates were recovered from blood culture while 20 % were recovered from deep wounds. The age of the patients included in the current study ranged from 55 to 85 years with a mean of 46.7 years. 56.7 % of the isolates were collected from female patients and 43.3% from male patients. As Regard isolate identification by Vitek-2C ANC ID CARD compared to MALDI-TOF (matrix-associated laser-induced desorption ionization-time of flight) as the gold standard method, the agreement was 86.7%. Regarding the antimicrobial susceptibility by disc diffusion method, all the isolates showed increased resistance against pencillin (100%), ciprofloxacin (70%), gentamicin (93.3%), tetracycline (76.6%), and clindamycin (70%). On the other hand, vancomycin and linezolid showed promising results where 100 % of the isolates were susceptible. Conclusion We spotlight that NDC spp. should no longer be considered contaminant bacteria in cultures, particularly in immunosuppressed patients. Timely identification, proper clinical correlation, and appropriate therapeutic intervention can lead to favorable outcomes. The high resistance to multiple classes of antibiotics as compounds of the MLSB group, fluoroquinolones, tetracyclines, and β- lactams, thus potentially limiting effective empirical treatment. However, vancomycin and linezolid are still effective against 100 % of the isolates.

Identification and antibiogram of clinically relevant non-diphtheriae Corynebacterium species

Identification and antibiogram of clinically relevant non-diphtheriae Corynebacterium species

2768. In Vitro Antibiotic Susceptibility Pattern of Non-diphtheriae Corynebacterium Isolates 2012-2023, reference lab experience

Abstract Background Non-diphtheriae Corynebacterium spp. are important human pathogens responsible for a broad spectrum of clinical syndromes. Data on species-specific susceptibility patterns is limited. In this study we aim to study these susceptibility patterns over time and summarize the species-specific resistance profile that could be used by clinicians to guide therapeutic decisions. Methods We conducted a retrospective review of all the Non-diphtheriae Corynebacterium isolates that were submitted for antimicrobial susceptibility testing (AST) between January-2012 and February of 2023 to our reference lab. Results A total of 1925 Corynebacterium isolates were submitted for susceptibility testing over the study period. The most commonly identified species was C. striatum (35.6%) followed by C. amycolatum (24.4%). Figure-1 outlines the species distribution. Temporal analysis showed decreasing susceptibility rates over the study period from 47.5% to 20.6% for penicillin & from 50% to 7.6% for clindamycin. Linezolid and vancomycin were universally active against all species (Table-1). Daptomycin non-susceptibility was reported in 25% of C. jeikeium isolates (MIC=2). High level resistance to daptomycin with MIC >256 were noted in seven C. striatum and single C. amycolatum isolate. Doxycycline has the highest susceptibility rates among the tested oral agents after linezolid for the non-striatum species. Significant variation in species-specific susceptibility pattern was noted and summarized in Table 1. C. striatum was associated with concerning resistance rates to beta-lactams, tetracyclines, trimethoprim-sulfamethoxazole & fluoroquinolones with limited reliable oral treatment options beside linezolid. C. kroppenstedtii (associated with granulomatous mastitis) showed high susceptibility rate to tetracyclines and trimethoprim-sulfamethoxazole despite prevalent penicillin resistance. Conclusion Non-diphtheriae Corynebacterium spp are commonly isolated from clinical specimens and are associated with various clinical syndromes. Significant variation in resistance profile exists between different species. Prevalent resistance among the most common species such as C. Striatum is associated with therapeutic challenges. Disclosures Christina G. Rivera (O'Connor), Pharm.D, Gilead Sciences: Advisor/Consultant|Gilead Sciences: Board Member|Gilead Sciences: Grant/Research Support|Gilead Sciences: Honoraria Audrey N. Schuetz, MD, Merck: Advisor/Consultant

Secondary Infections in Patients with Extremely Severe COVID-19 During ECMO Therapy

Up to 70% of patients hospitalized for COVID-19 need respiratory support, up to 10% need high-flow oxygen therapy, non-invasive and invasive ventilation. However, standard methods of respiratory support are ineffective in 0.4-0.5% of patients. In case of potentially reversible critical refractory respiratory failure that patients may require ECMO. Management of patients with extremely severe COVID-19 associates with numerous clinical challenges, including critical illness, multiple organ dysfunction, blood coagulation disorders, requiring prolonged ICU stay and care, use of multiple pharmacotherapies including immunosuppressive drugs. Pharmacological suppression of immunity is associated with a significant increase in the risk of secondary bacterial and fungal infections. Currently, data on epidemiology of secondary infections in patients with COVID-19 undergoing ECMO is limited.Aim. To study the prevalence and etiology of secondary infections associated with positive blood cultures in patients with extremely severe COVID-19 requiring ECMO.Materials and methods. A single-center retrospective non-interventional epidemiological study including 125 patients with extremely severe COVID-19 treated with ECMO in April 2020 to December 2021.Results. Out of 700 blood culture tests performed in 125 patients during the study, 250 tests were positive confirming bacteremia/fungemia. Isolated pathogens varied depending on the duration of ECMO: gram-positive bacteria (primarily coagulase-negative staphylococci) dominated from the initiation of ECMO support; increased duration of ECMO associated with an increasing the proportion of pathogens common in ICU (Klebsiella pneumoniae and/or Acinetobacter baumannii with extensively drug resistant and pan-drug resistant phenotypes, and vancomycin-resistant Enterococcus faecium). When ECMO lasted more than 7-14 days, opportunistic pathogens (Candida species, Stenotrophomonas maltophilia, Providencia stuartii, non-diphtheria corynebacteria, Burkholderia species and others) prevailed as etiological agents.Conclusion. Longer duration of ECMO resulted in increasing the rates of infectious complications. In patients undergoing ECMO for more than 14 days, the microbiological landscape becomes extremely diverse, which hampers choosing an empirical antimicrobial therapy. Since potential pathogens causing secondary infections in patients during ECMO are difficult to predict, rapid identification of rare opportunistic pathogens and their sensitivity profile, followed by targeted administration of antimicrobials, seems most beneficial.

Corynebacterium guaraldiae sp. nov.: a new species of Corynebacterium from human infections.

Non-diphtheria Corynebacterium species (NDC) belonging to the human skin and mucosa microbiota are frequently neglected as contaminants. However, reports of human infections by Corynebacterium spp. have increased considerably in recent years. In this study, a group of six NDC isolates of urine (n = 5) and sebaceous cyst (n = 1) from two South American countries were identified at genus level or misidentified based on API® Coryne and genetic/molecular analyses. The 16S rRNA (99.09-99.56%) and rpoB (96.18-97.14%) gene sequence similarities of the isolates were higher when compared with Corynebacterium aurimucosum DSM 44532T. Multilocus sequence analysis (MLSA) indicated that these six NDC isolates compose a distinctive phylogenetic clade. Genome-based taxonomic analysis with the whole-genome sequences was able to separate these six isolates from other known Corynebacterium type strains. Average nucleotide identity (ANI), average amino acid identity (AAI), and digital DNA-DNA hybridization (dDDH) values between closely related type strains and the six isolates were considerably lower than the currently recommended threshold values for species circumscription. Phylogenetic and genomic taxonomy analyses indicated these microorganisms as a novel Corynebacterium species, for which we formally propose the name Corynebacterium guaraldiae sp. nov. with isolate 13T (= CBAS 827T = CCBH 35012T) as type strain.

Re-identification and Characterization of Multidrug-Resistant Corynebacterium haemomassiliense-Like Organisms from Humans

Recently, non-diphtheriae Corynebacterium spp. have been increasingly reported in patients. In addition, several novel species of Corynebacterium isolated from humans. Here, we report two cases of human infections caused by Corynebacterium haemomassiliense-like organisms, which had not been identified at the species level by MALDI-TOF MS analysis. They were revealed to be closely related to C. haemomassiliense, a recently described species by three housekeeping genes (16S rRNA, rpoB, and gyrA) and phenotypic features. Both strains were multidrug-resistant but susceptible to vancomycin, meropenem, and linezolid. Our report suggests that human infections by the recently described Corynebacterium species may not be limited to a specific region, in addition to difficulty of classifying the genus Corynebacterium.

Relapsing and refractory peritoneal dialysis peritonitis caused by Corynebacterium amycolatum

BackgroundPeritonitis is an important complication and cause of morbidity in patients undergoing peritoneal dialysis (PD). Corynebacterium species, often considered skin and mucosal contaminants, are a rare cause of PD-associated peritonitis and have been acknowledged in published guidelines for the diagnosis and treatment of PD peritonitis only over the last decade.Case-Diagnosis/TreatmentWe present two children with difficult-to-treat episodes of PD peritonitis due to Corynebacterium amycolatum. Episodes were associated with fever, abdominal pain and cloudy dialysate, high dialysate polymorphonuclear leukocyte counts, and elevated serum C-reactive protein and procalcitonin concentrations. Symptoms persisted beyond 5 days in 4 of 5 peritonitis episodes, and peritonitis relapsed despite in vitro sensitivity of the bacterial isolates to guideline-recommended antibiotics. C. amycolatum was cultured from the PD catheter tip despite 4 weeks of intraperitoneal glycopeptide therapy and clinical peritonitis resolution suggestive of efficient biofilm formation. Our systematic literature search identified three previous (adult) case descriptions of C. amycolatum peritonitis, all with repeat episodes by the same organism. The incidence of C. amycolatum as a cause of PD peritonitis has not yet been established but is likely underreported due to challenges in species differentiation.ConclusionsC. amycolatum is a rarely identified cause of refractory and/or relapsing PD peritonitis. Species differentiation of non-diphtheriae Corynebacterium isolates is critical, and prolonged antibiotic treatment, preferably with a glycopeptide antibiotic, is recommended, with a low threshold for PD catheter change or removal in case of repeat peritonitis.

In vitro Studies of Non-Diphtheriae Corynebacterium Isolates on Antimicrobial Susceptibilities, Drug Resistance Mechanisms, and Biofilm Formation Capabilities.

ObjectiveThis study aimed to investigate the antimicrobial susceptibilities, drug resistance mechanisms, and biofilm formation capacities of non-diphtheriae Corynebacterium strains isolated from sterile midstream urine of hospitalized patients with clinical urinary tract infections (UTIs).MethodsA total of 45 non-diphtheriae Corynebacterium isolates were recovered from sterile midstream urine. The available data of 45 patients were collected. Minimum inhibitory concentrations (MICs) of 10 commonly used antibiotics were determined. Meanwhile, the molecular resistance mechanisms of each agent were performed through PCR with specific primers. Moreover, the biofilm formation capability of each isolate on abiotic surfaces was detected with the MTT method.ResultsIn this study, the most prevalent three species were C. striatum (15/45, 33.3%), C. glucuronolyticum (9/45, 20.0%) and C. urealyticum (8/45, 17.8%). These three species also accounted for most renal and ureteral calculi cases. Male patients older than 50 years, especially those with underlying diseases, were more susceptible to non-diphtheriae Corynebacterium infection. All the 45 isolates were 100% susceptible to vancomycin and linezolid, but highly resistant to macrolide–lincosamide–streptogramin B (MLSB), fluoroquinolones, tetracyclines and β-lactams with corresponding mechanisms. The detection rate of multidrug–resistant (MDR) non-diphtheriae Corynebacterium is 91.1%. All isolates are able to form biofilm on abiotic surfaces, except those of C. urealyticum, C. tuberculostearicum and C. jeikeium. Isolates of C. glucuronolyticum and C. Striatum possessed the strongest biofilm formation capacity. C. amycolatum could form biofilm, but varied greatly among different isolates.ConclusionC. striatum, C. glucuronolyticum and C. urealyticum were the most prevalent species relevant to UTIs. The high occurrence of MDR isolates and high diversities in resistance profiles, and the distinctive abilities of biofilm formation highlighted the urgency for identification to species level. We should pay more attention to the drug resistance profiles of non-diphtheriae Corynebacterium, which would help improve empirical antibiotic therapy and reduce drug resistance transmission.

Non-diphtheriae Corynebacteria Causing Breast Abscess in Non-lactating Females Seeking to Avoid Pitfalls in Diagnosis of Chronic Breast Infections Through Diagnostic Stewardship

Background: Consideration of non-diphtheriae Corynebacteria as an infective organism in mastitis, is usually neglected. So, infections due to Corynebacterium species may remain undiagnosed or misdiagnosed. The aim of our study was to focus on non-diphtheriae Corynebacterium as causative organisms in breast infections using scientific logic and technology of Matrix Assisted Laser Desorption Ionisation Time of Flight Mass Spectrometry (MALDITOF MS) to categorize the isolates upto species level. The clinical correlation and response to management especially targeted antimicrobials was also recorded.
 Methods: All the consecutive pus/tissue samples from Breast abscess received during study period were processed as per the standard guidelines. The identification of the isolate was done by automated methods. The cytopathological/histopathological and clinical details of the patients with infection due to Corynebacterium sp. were recorded and analyzed.
 Results: Out of 52 non-duplicate samples, five showed growth of non-diphtheriae Corynebacteria. These were identified as C. kroppenstedtii and C. amycolatum. The antimicrobial susceptibility testing showed 100% susceptibility to Amoxicillin-clavulanate, Tetracycline, Vancomycin and Linezolid for all the five isolates. Histopathological examination was suggestive of chronic inflammatory mastitis/Granulomatous mastitis.
 Conclusion: Non diphtheriae Corynebacteria particularly lipophilic Corynebacteria have a predilection to cause infections of breast tissue and Breast abscess which may mimic as tubercular abscess or chronic non-specific mastitis leading to unnecessary usage of antimicrobials. So, these isolates from breast tissue/pus should not be ignored rather definitely identified. This would also ensure diagnostic stewardship.

Virulence potential of the first Corynebacterium mycetoides strain isolated from human urine: a rare species of Corynebacterium

For many years, the potential pathogenic of non-diphtheriae corynebacteria were underestimated. Nowadays, a growing number of Corynebacterium species are recognized as opportunistic agents of human infections, mainly in hospital settings. In addition, multidrug-resistant Corynebacterium isolates from clinical specimens, have been reported and the role of Corynebacterium spp. in urinary tract infections (UTIs) has been highlighted. Several studies have reported Corynebacterium species as the agent of UTIs especially in patients with risk factors. Thus, the present work aimed to report the first isolation of Corynebacterium mycetoides from human urine and an initial study on its virulence properties. The isolate, initially characterized by phenotypical tests as a multidrug-resistant Corynebacterium sp., was recovered from the urine of a female transplant patient. Mass spectrometry and 16S rRNA and rpoB genes sequencing identified the isolate as C. mycetoides. The isolate was found able to adhere to and survive into epithelial cells (Vero cells), and its pathogenic potential was confirmed when tested against Caenorhabditis elegans nematode. The results obtained suggest that C. mycetoides is a potential pathogen for the urinary tract in humans and for a better understanding of the multifactorial mechanisms of virulence, studies about this species should be continued.

Lateral Flow Immunoassay for Rapid Identification of the Diphtheria Agent

<h3>Purpose</h3> Recent reports from several EU countries indicate circulation of toxigenic <i>Corynebacterium diphtheriae</i> among refugees from Asia and Africa as well as children's deaths among indigenous populations. Toxigenic <i>C. diphtheriae</i> carriers pose a serious risk for unvaccinated persons and ones with low level of immunological protection, the number of which in Europe has increased. The International system of quality control revealed the poor state of laboratory diagnosis of diphtheria in EU. This is mostly due to the absence of simple, standard and reliable method of the diphtheria toxin (DT) detection. The common method for detecting the production of DT by corynebacteria, gel immunoprecipitation or Elek test is vulnerable, time consuming and laborious. PCR for DT gene detection cannot be used instead of the Elek test, since non-toxigenic toxin gene-bearing (NTTB) <i>C. diphtheriae</i> and <i>C. ulcerans</i> circulate worldwide. The aim of this study was development and evaluation of rapid immunochemical test for the detection of DT. <h3>Methods & Materials</h3> Lateral Flow Immunoassay (LFIA) strips were designed using monoclonal antibodies against DT. Two hundred and fifty of toxigenic, non-toxigenic <i>C. diphtheriae, C. ulcerans</i> and other corynebacteria isolated in Germany in 2019-2021 were tested for DT after 6 hours of growth on the WHO-recommended liquid nutrient medium. <h3>Results</h3> A complete convergence of the conventional method of DT detection - Elek test and the LFIA for all 250 isolates of diphtheria and non-diphtheria corynebacteria was observed. The use of the LFIA makes it possible to significantly simplify DT detection and reduce the time of analysis from 24-48 to 4-6 hours. Thus, the possibility of replacing the Elek test with the LFIA appears real. <h3>Conclusion</h3> The availability of such a simple and reliable method for DT detection will speed up and increase the accuracy of diphtheria diagnosis globally. The efficient diphtheria laboratory diagnosis will enable timely initiation of anti-diphtheritic treatment that will save lives, prevent the development of complications, as well as stop the spread of the disease through early initiation of preventive measures.

Non-diphtheriae Corynebacterium species are associated with decreased risk of pneumococcal colonization during infancy.

Streptococcus pneumoniae (pneumococcus) is a leading cause of severe infections among children and adults. Interactions between commensal microbes in the upper respiratory tract and S. pneumoniae are poorly described. In this study, we sought to identify interspecies interactions that modify the risk of S. pneumoniae colonization during infancy and to describe development of the upper respiratory microbiome during infancy in a sub-Saharan African setting. We collected nasopharyngeal swabs monthly (0-6 months of age) or bimonthly (6-12 months of age) from 179 mother-infant dyads in Botswana. We used 16S ribosomal RNA gene sequencing to characterize the nasopharyngeal microbiome and identified S. pneumoniae colonization using a species-specific PCR assay. We detect S. pneumoniae colonization in 144 (80%) infants at a median age of 71 days and identify a strong negative association between the relative abundance of the bacterial genera Corynebacterium within the infant nasopharyngeal microbiome and the risk of S. pneumoniae colonization. Using in vitro cultivation experiments, we demonstrate growth inhibition of S. pneumoniae by secreted factors from strains of several Corynebacterium species isolated from these infants. Finally, we demonstrate that antibiotic exposures and the winter season are associated with a decline in the relative abundance of Corynebacterium within the nasopharyngeal microbiome, while breastfeeding is associated with an increase in the Corynebacterium relative abundance. Our findings provide novel insights into the interspecies interactions that contribute to colonization resistance to S. pneumoniae and suggest that the nasopharyngeal microbiome may be a previously unrecognized mechanism by which environmental factors influence the risk of pneumococcal infections during childhood. Moreover, this work lays the foundation for future studies seeking to use targeted manipulation of the nasopharyngeal microbiome to prevent infections caused by S. pneumoniae.

Direct detection of Corynebacterium striatum, Corynebacterium propinquum, and Corynebacterium simulans in sputum samples by high-resolution melt curve analysis

BackgroundPulmonary infections caused by non-diphtheriae corynebacteria are increasing. However, rapid identification of Corynebacterium species poses a challenge due to the low genetic variation within the genus.MethodsThree reference strains and 99 clinical isolates were used in this study. A qPCR followed by high-resolution melting (HRM) targeting ssrA was performed to simultaneously identify C. striatum, C. propinquum and C. simulans. To further evaluate this assay’s performance, 88 clinical sputum samples were tested by HRM and the detection results were compared with those of the traditional culture method and multiple cross-displacement amplification (MCDA) assay.ResultsThe melting curve produced by a pair of universal primers generated species-specific HRM curve profiles and could distinguish the three target species from other related bacteria. The limit of detection of HRM assay for DNA from the three purified Corynebacterium species was 100 fg. Compared with the culture method, HRM detected 22 additional positive specimens, representing a 23.9% relative increase in detection rate. The HRM assay had 98.4% (95% confidence interval [CI], 90.5–99.9%) sensitivity and 100% (95% CI, 82.8–100%) specificity. Additionally, 95.5% concordance between HRM and MCDA (κ = 0.89 [95% CI, 0.79–0.99]) was noted.ConclusionsThe HRM assay was a simple, rapid, sensitive, and specific diagnostic tool for detecting C. striatum, C. propinquum, and C. simulans, with the potential to contribute to early diagnosis, epidemiological surveillance, and rapid response to outbreak.

Kan Kültürlerinden İzole Edilen Difteri-Dışı Corynebacterium Suşlarının Biyofilm Oluşturması ve Antimikrobiyal Duyarlılıkları: Türkiye’deki ilk Bildirim

ObjectiveNon-diphtheriae Corynebacterium strains have been recognized as important pathogens after decades of confusion regarding their microbiological classification and clinical significance. The aim of this study was to identify non-diphtheriae Corynebacterium strains and the prevalence of biofilm formation and antimicrobial resistance.MethodIn total, 126 non-diphtheriae Corynebacterium strains were isolated from blood cultures of inpatients with bacteremia in our hospital between January 2015 and January 2020. Blood cultures were analyzed with the Bactec-9120 system. Strains were identified using MALDI-TOF MS (Bruker Daltonics, Germany). Antimicrobial susceptibilities were determined using the Kirby-Bauer disk diffusion method on a Mueller-Hinton agar and evaluated according to EUCAST standards. Biofilm formation was assessed with the Congo Red Agar method.ResultsCorynebacterium striatum and Corynebacterium matruchotii were the most prevalent with 29 and 26 isolates, respectively. Biofilm production was detected in 62.06% (18/29) of C. striatum, in 53.8% (14/26) of C. matruchotii, in 50% (9/18) of Corynebacterium afermentans, 50% (6/12) of Corynebacterium amycolatum, and in 46% (7/15) of Corynebacterium jeikeium strains. Among the five most prevalent strains, we found a high biofilm rate of 54%. The resistance rates to penicillin, clindamycin, ciprofloxacin, rifampicin, tetracycline, and gentamicin were 91.2%, 87.3%, 79.3%, 56.3%, 45.2%, and 39.6%, respectively. All 126 strains were susceptible to vancomycin and linezolid.ConclusionNon-diphtheriae Corynebacterium strains isolated from blood cultures of hospitalized patients with bacteremia may have multidrug resistance and the ability to produce biofilm. These results emphasize the importance of identifying strains and determining their antimicrobial susceptibility and biofilm production potential.

Antimicrobial Susceptibility Testing for Corynebacterium Species Isolated from Clinical Samples in Romania

Antimicrobial resistance is one of the most important public health issues. Besides classical multidrug resistance species associated with medical care involved in superficial or invasive infections, there are strains less commonly associated with hospital or outpatient setting’s infections. Non-diphtheria Corynebacterium spp. could produce infections in patients with or without immune-compromised status. The aim of our study was to determine the susceptibility to antimicrobial agents to Corynebacterium spp. from clinical samples collected from Romanian hospitalized individuals and outpatients. Twenty Corynebacterium strains were isolated and identified as Corynebacterium striatum (n = 7), Corynebacterium amycolatum (n = 7), C. urealyticum (n = 3), Corynebacterium afermentans (n = 2), and Corynebacterium pseudodiphtheriticum (n = 1). All isolates have been tested for antibiotic susceptibility by standardized disc diffusion method and minimal inhibitory concentration (MIC) tests. Seventeen isolates demonstrated multidrug resistance phenotypes. The molecular support responsible for high resistance to quinolones for ten of these strains was determined by the detection of point mutation in the gene sequence gyrA.

Cypopathic effect of diphtheria pathogen in the composition of biofilm.

When the nasopharynx is colonized with toxigenic strains of the diphtheria pathogen, toxin is released, which contributes to the death of epithelial cells. But in bacterial carriers, the development of the clinical picture of the disease does not occur. This is due to the peculiarities of the state of their immune system, as well as the peculiarities of the production of diphtheria exotoxin by corynebacteria in the biofilm. Goal. Determining the nature of the cytopathic effect of C. diphtheriae as part of a biofilm in CHO-K1 cell culture. The planktonic and biofilm (120- and 720-hour) cultures of the strains were studied: C. diphtheriae gravis tox+ № 665, C. diphtheriae gravis tox+ № 6765, C. diphtheriae mitis tox+ № 269, C. diphtheriae gravis tox+ isolated from a patient with a diagnosis Localized oropharyngeal diphtheria C. diphtheriae gravis with a silent tox-gene. Biofilm (120- and 720-hour) cultures of diphtheria pathogen strains were obtained according to the Watnik method. The cytopathic effect of corynebacterial strains was studied on a CHO-K1 cell culture, taking into account in an inverted microscope. When studying the cytopathic effect of planktonic cultures of toxigenic strains of corynebacteria, it was found that the number of living CHO-K1 cells after 24 hours was insignificant (25.3±1.2%) and sharply decreased (2.5±0.5%) after 72 hours of cultivation. Under the influence of biofilm and, especially, 720-hour cultures, a different cytopathic effect dynamics was found: the number of living cells after 24 hours remained significant (82.5±2.2%), while at 72-hour it decreased to 25.0±3.0%. In the study of filtrates of planktonic and biofilm cultures of C. diphtheriae strain with a «silent» tox-gene, similar patterns were revealed. However, the number of live CHO-K1 cells when exposed to the filtrate of a 720-hour biofilm culture was significantly higher (p≤0.05) than when studying toxigenic strains of corynebacteria. Considering the nature of the cytopathic action, it was found that planktonic cultures of toxigenic strains of corynebacteria are characterized by a change in the cell monolayer, manifested by their thinning and elongation. The study of 720-hour biofilm cultures at 72-hour exposure revealed the appearance of a large number of rounded cells (63-69%). The cytopathic effect, formed under the influence of filtrates of planktonic and biofilm cultures of C. diphtheriae with a «silent» tox-gene, as well as strains of non-diphtheria corynebacteria, is characterized by rounding of cells and the formation of symplasts. In the biofilm, the intensity of the cytopathic effect of toxigenic C. diphtheriae strains and C. diphtheriae strain with a silent tox-gene decreased. CPD, manifested by thinning and lengthening of CHO-K1 cells, is associated with the action of diphtheria exotoxin, and rounding is associated with corynebacterial enzymes and, apparently, fragments of surface structures - adhesins. Decreased release of toxin and enzymes beyond the C. bihfilm matrix is a significant cause of the «asymptomatic» carriage of diphtheria.

Cytokine Levels in the In Vitro Response of T Cells to Planktonic and Biofilm Corynebacterium amycolatum

Unravelling of the interplay between the immune system and non-diphtheria corynebacteria would contribute to understanding their increasing role as medically important microorganisms. We aimed at the analysis of pro- (TNF, IL-1β, IL-6, IL-8, and IL-12p70) and anti-inflammatory (IL-10) cytokines produced by Jurkat T cells in response to planktonic and biofilm Corynebacterium amycolatum. Two reference strains: C. amycolatum ATCC 700207 (R-CA), Staphylococcus aureus ATCC 25923 (R-SA), and ten clinical strains of C. amycolatum (C-CA) were used in the study. Jurkat T cells were stimulated in vitro by the planktonic-conditioned medium (PCM) and biofilm-conditioned medium (BCM) derived from the relevant cultures of the strains tested. The cytokine concentrations were determined in the cell culture supernatants using the flow cytometry. The levels of the cytokines analyzed were lower after stimulation with the BCM when compared to the PCM derived from the cultures of C-CA; statistical significance (p < 0.05) was observed for IL-1β, IL-12 p70, and IL-10. Similarly, planktonic R-CA and R-SA stimulated a higher cytokine production than their biofilm counterparts. The highest levels of pro-inflammatory IL-8, IL-1β, and IL-12p70 were observed after stimulation with planktonic R-SA whereas the strongest stimulation of anti-inflammatory IL-10 was noted for the BCM derived from the mixed culture of both reference species. Our results are indicative of weaker immunostimulatory properties of the biofilm C. amycolatum compared to its planktonic form. It may play a role in the persistence of biofilm-related infections. The extent of the cytokine response can be dependent on the inherent virulence of the infecting microorganism.

The Association Between the Developing Nasal Microbiota of Hospitalized Neonates and Staphylococcus aureus Colonization.

BackgroundHospitalized neonates are at high risk for invasive Staphylococcus aureus infections. S. aureus nasal colonization often precedes infection. The nasal microbiota may preclude or support colonization. We aimed to characterize and compare the nasal microbiota of hospitalized neonates who acquire S. aureus colonization (cases) and those who do not acquire S. aureus (controls).MethodsWe obtained residual nares samples from hospitalized neonates who were screened weekly for S. aureus nasal colonization and treated with intranasal mupirocin if colonized. Eight cases were matched based on chronologic age and systemic antibiotic exposure to 7 controls. We extracted DNA, sequenced the V3-V4 region of the 16s rRNA gene, and performed taxonomic assignments. The bacterial species richness, relative abundance, and in silico predicted gene content were compared between cases and controls at 7 days before S. aureus acquisition, at the time of acquisition, and 7 days after acquisition and treatment.ResultsCommon commensals including nondiphtheriae corynebacteria were more abundant in the nares of controls and Rothia mucilaginosa was more abundant in cases 7 days after intranasal mupirocin treatment than in cases 7 days before S. aureus acquisition. Controls and treated cases had a higher predicted abundance of genes contributing to the synthesis of certain antimicrobial compounds than in cases before S. aureus acquisition.ConclusionsNeonates without S. aureus nasal colonization had a higher abundance of bacterial species that antagonize S. aureus directly or by selecting for beneficial co-colonizers. These differences may inform novel S. aureus infection prevention strategies in high-risk infants.

Corynebacteria as a cause of pulmonary infection: a case series and literature review

BackgroundIn most cases of community-acquired pneumonia (CAP), an etiologic agent is not determined; the most common report from the microbiological evaluation of sputum cites “normal respiratory flora.” Non-diphtheria Corynebacterium spp., a component of this flora, is commonly viewed as a contaminant, but it may be the cause of pneumonia and the frequency with which it causes CAP may be underestimated.Case presentationsThis report present 3 cases of CAP in which Corynebacterium spp. was clearly the predominant isolate; identification was confirmed by matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry. Two cases were caused by C. propinquum and one by C. striatum. Two patients had a tracheostomy and one was on hemodialysis. Patients who received an appropriate antibiotic responded well.ConclusionWhen identified as the predominant isolate in sputum from a patient with CAP, Corynebacterium spp. should be considered as a potential cause of the infection. In cases with patients who have compromised airway clearance or who are immunocompromised, microaspiration may be responsible. While some Corynebacterium spp. are suspectible to antibiotics usually prescribed for CAP, others are susceptible only to vancomycin or aminoglycosides. Vancomycin is thus the appropriate empiric antibiotic, pending speciation and susceptibility test results. The number of reported cases with result of antibiotic susceptibility testing, however, remains limited, and further investigation is needed. Non-diphtheria Corynebacterium spp. represent a noteworthy clinical cause of pneumonia. Identification by Gram stain and as a predominant organism on culture demands careful consideration for management.