Corynebacterium Ulcerans Strains Research Articles

Complete Genome Sequence of Corynebacterium ulcerans Strain TSU-28, Harboring Two Diphtheria Toxin Genes, Isolated from a Patient with Diphtheria-Like Symptoms.

Diphtheria toxin-producing Corynebacterium ulcerans is an emerging zoonotic pathogen that causes severe disease in humans. Here, we report the complete genome sequence of C. ulcerans strain TSU-28, harboring two diphtheria toxin genes, which was isolated from the throat of a patient with diphtheria-like symptoms in 2019 in Japan.

Draft Genome Sequences of Six Corynebacterium ulcerans Strains Isolated from Humans and Animals in Austria, 2013 to 2019.

Corynebacterium ulcerans is an emerging pathogen responsible for severe diseases in humans and animals. Here, we present the draft genome of six C. ulcerans strains isolated in Austria. These draft genomes have 2,446,822 to 2,551,141 bp encoding 57 to 60 RNAs.

Induction of Necrosis in Human Macrophage Cell Lines by Corynebacterium diphtheriae and Corynebacterium ulcerans Strains Isolated from Fatal Cases of Systemic Infections.

When infecting a human host, Corynebacterium diphtheriae and Corynebacterium ulcerans are able to impair macrophage maturation and induce cell death. However, the underlying molecular mechanisms are not well understood. As a framework for this project, a combination of fluorescence microscopy, cytotoxicity assays, live cell imaging, and fluorescence-activated cell sorting was applied to understand the pathogenicity of two Corynebacterium strains isolated from fatal cases of systemic infections. The results showed a clear cytotoxic effect of the bacteria. The observed survival of the pathogens in macrophages and, subsequent, necrotic lysis of cells may be mechanisms explaining dissemination of C. diphtheriae and C. ulcerans to distant organs in the body.

Beyond diphtheria toxin: cytotoxic proteins of Corynebacterium ulcerans and Corynebacterium diphtheriae.

Diphtheria toxin is one of the best investigated bacterial toxins and the major virulence factor of toxigenic Corynebacterium diphtheriae and Corynebacterium ulcerans strains. However, also diphtheria toxin-free strains of these two species can cause severe infections in animals and humans, indicating the presence of additional virulence factors. In this study, we present a first characterization of two proteins with cytotoxic effect in corynebacteria. A putative ribosome-binding protein (AEG80717, CULC809_00177), first annotated in a genome sequencing project of C. ulcerans strain 809, was investigated in detail together with a homologous protein identified in C. diphtheriae strain HC04 (AEX80148, CDHC04_0155) in this study. The corresponding proteins show striking structural similarity to Shiga-like toxins. Interaction of wild-type, mutant and complementation as well as overexpression strains with invertebrate model systems and cell lines were investigated. Depending on the presence of the corresponding genes, detrimental effects were observed in vivo in two invertebrate model systems, Caenorhabditis elegans and Galleria mellonella, and on various animal and human epithelial and macrophage cell lines in vitro. Taken together, our results support the idea that pathogenicity of corynebacteria is a multifactorial process and that new virulence factors may influence the outcome of potentially fatal corynebacterial infections.

Genomic analyses reveal two distinct lineages of Corynebacterium ulcerans strains.

Corynebacterium ulcerans is an important zoonotic pathogen which is causing diphtheria-like disease in humans globally. In this study, the genomes of three recently isolated C. ulcerans strains, 4940, 2590 and BR-AD 2649, respectively from an asymptomatic carrier, a patient with pharyngitis and a canine host, were sequenced to investigate their virulence potential. A comparative analysis was performed including the published genome sequences of 16 other C. ulcerans isolates. C. ulcerans strains belong to two lineages; 13 strains are grouped together in lineage 1, and six strains comprise lineage 2. Consistent with the zoonotic nature of C. ulcerans infections, isolates from both the human and canine hosts clustered in both the lineages. Most of the strains possessed spaDEF and spaBC gene clusters along with the virulence genes cpp, pld, cwlH, nanH, rpfI, tspA and vsp1. The gene encoding Shiga-like toxin was only present in one strain, and 11 strains carried the tox gene encoding the diphtheria-like toxin. However, none of strains 4940, 2590 and BR-AD 2649 carried any toxin genes. These strains varied in the number of prophages in their genomes, which suggests that they play an important role in introducing diversity in C. ulcerans. The pan-genomic analyses revealed a variation in the number of membrane-associated and secreted proteins that may contribute to the variation in pathogenicity among different strains.

Surface and Extracellular Proteome of the Emerging Pathogen Corynebacterium ulcerans.

Corynebacterium ulcerans is an emerging pathogen, which is increasingly recognized as an etiological agent of diphtheria, but can also evoke ulcers of the skin and systemic infections in humans. Besides man, the bacteria can colonize a wide variety of different animals, including cattle and pet animals, which might serve as a reservoir for human infections. In this study, surface-located proteins and the exoproteome of two Corynebacterium ulcerans strains were analyzed, since these may have key roles in the interaction of the pathogen with host cells. Strain 809 was isolated from a fatal case of human respiratory tract infection, while strain BR-AD22 was isolated from a nasal swap of an asymptomatic dog. While a very similar pattern of virulence factors was observed in the culture supernatant and surface protein fractions of the two strains, proteome analyses revealed a higher stability of 809 cells compared to strain BR-AD22. During exponential growth, 17% of encoded proteins of strain 809 were detectable in the medium, while 38% of the predicted proteins encoded by the BR-AD22 chromosome were found. Furthermore, the data indicate differential expression of phospholipase D and a cell wall-associated hydrolase, since these were only detected in strain BR-AD22.

Outbreak with clonally related isolates of Corynebacterium ulcerans in a group of water rats.

BackgroundThe zoonotic bacterium Corynebacterium ulcerans may be pathogenic both in humans and animals: toxigenic strains can cause diphtheria or diphtheria-like disease in humans via diphtheria toxin, while strains producing the dermonecrotic exotoxin phospholipase D may lead to caseous lymphadenitis primarily in wild animals. Diphtheria toxin-positive Corynebacterium ulcerans strains have been isolated mainly from cattle, dogs and cats.ResultsHere, we report a series of ten isolations of Corynebacterium ulcerans from a group of water rats (Hydromys chrysogaster) with ulcerative skin lesions, which were kept in a zoo. The isolates were clearly assigned to species level by biochemical identification systems, Fourier-transform infrared-spectroscopy, Matrix-assisted laser desorption/ionization-time of flight mass spectrometry and partial rpoB sequencing, respectively. All ten isolates turned out to represent the same sequence type, strongly indicating a cluster of infections by clonally-related isolates as could be demonstrated for the first time for this species using multilocus sequence typing. Unequivocal demonstration of high relatedness of the isolates could also be demonstrated by Fourier-transform infrared-spectroscopy. All isolates were lacking the diphtheria toxin encoding tox-gene, but were phospholipase D-positive.ConclusionsOur results indicate that water rats represent a suitable new host species that is prone to infection and must be regarded as a reservoir for potentially zoonotic Corynebacterium ulcerans. Furthermore, the applied methods demonstrated persistent infection as well as a very close relationship between all ten isolates.

Corynebacterium ulceransin Ferrets

<i>Corynebacterium ulcerans</i>in Ferrets

Rapid Detection and Molecular Differentiation of Toxigenic Corynebacterium diphtheriae and Corynebacterium ulcerans Strains by LightCycler PCR

The systemic symptoms of diphtheria are caused by the tox-encoded diphtheria toxin (DT) which is produced by toxigenic Corynebacterium spp. Besides the classical agent C. diphtheriae, the zoonotic pathogen C. ulcerans has increasingly been reported as an emerging pathogen for diphtheria. The reliable detection of toxigenic Corynebacterium spp. is of substantial importance for both diphtheria surveillance in the public health sector and the clinical workup of a patient with diphtherialike symptoms. Since the respective tox genes of C. diphtheriae and C. ulcerans differ from each other in both DNA and amino acid sequence, both tox genes should be covered by novel real-time PCR methods. We describe the development and validation of a LightCycler PCR assay which reliably recognizes tox genes from both C. diphtheriae and C. ulcerans and differentiates the respective target genes by fluorescence resonance energy transfer (FRET) hybridization probe melting curve analysis.

Corynebacterium ulcerans from Diseased Wild Boars

Two Corynebacterium strains were isolated from lymph nodes of wild boars showing severe alterations caused by caseous lymphadenitis. The wild boars came from different districts in southern Germany; one was found dead, the other had been shot. The two Corynebacterium strains obtained were both positive for phospholipase D. Further analysis of biochemical profiles did not allow unambiguous differentiation between Corynebacterium ulcerans and Corynebacterium pseudotuberculosis. Fourier-transformed infrared spectroscopy as well as partial sequencing of the genes for 16S rRNA and RNA polymerase beta subunit (rpoB) clearly identified both strains as Corynebacterium ulcerans. The tox gene for diphtheria toxin (DT) could be detected in both porcine isolates by PCR. Partial DNA sequencing of this tox gene showed significant differences from sequences described for other Corynebacterium ulcerans strains and a higher degree of similarity to that of Corynebacterium diphtheria. Production of diphtheria toxin could not be detected. These results indicate that wild game could be a reservoir for zoonotic Corynebacterium ulcerans.

Detection of ToxigenicCorynebacterium diphtheriaeandCorynebacterium ulceransStrains by a Novel Real-Time PCR

Diphtheria is caused by both toxigenic Corynebacterium diphtheriae and Corynebacterium ulcerans strains harboring lysogenic, tox -bearing beta-corynephages. While C. diphtheriae has caused most diphtheria cases for decades, the emerging pathogen C. ulcerans is becoming the major cause for diphtheria

Characterization of toxigenic Corynebacterium ulcerans strains isolated from humans and domestic cats in the United Kingdom.

In the United Kingdom there has been a marked increase in the number of human infections caused by toxigenic Corynebacterium ulcerans. During 2002 and 2003 the organism was also isolated from several domestic cats with bilateral nasal discharge. As C. ulcerans has never previously been isolated from cats, the 16S rRNA gene from three cat isolates was sequenced to confirm their species identities. Fifty clinical isolates from the United Kingdom isolated from 1986 to 2003 and seven cat isolates were characterized by ribotyping to determine whether the ribotypes of the cat isolates were genotypically related to those found for human clinical isolates. For comparison, the genotypes of 11 overseas isolates and 13 isolates from H. R. Carne's collection isolated between 1933 and 1979 were also determined. Strains isolated from domestic cats were found to exhibit the predominant ribotypes observed among human clinical isolates, suggesting that C. ulcerans isolated from cats could be a potential reservoir for human infection.

A modified Elek test for detection of toxigenic corynebacteria in the diagnostic laboratory

The detection of toxigenicity among Corynebacterium diphtheriae and Corynebacterium ulcerans strains is the most important test for the microbiological diagnosis of diphtheria. Difficulties with current methods, in particular the Elek test, are well documented. We therefore describe a modified Elek test which provides an accurate result after only 16 h of incubation, in contrast to 48 h for the conventional test.

Purification and characterization of a Corynebacterium ulcerans bacteriocin (ulceracin 378).

Corynebacterium ulcerans strain 378 produces a bacteriocin (ulceracin 378) and a toxin when grown on semi-solid medium. Ulceracin 378 was purified 360-fold by dialysis and chromatography on DEAE-cellulose and Sephadex G-200. On the basis of Ultrogel AcA22 gel filtration its molecular weight was about 900 000. It could be dissociated by 2-mercaptoethanol and sodium dodecyl sulphate into smaller subunits of 25 000. The bactericidal activity was associated with this subunit which contained no carbohydrate or lipid. Ulceracin 378 was thermostable and stable over a wide pH range. Purified ulceracin 378 did not have a toxic effect (lethal) on guinea-pigs and rabbits and was immunologically distinct from the toxin.

Proof for the lack of diphtheria toxin in culture filtrates of corynebacterium ulcerans strain ATCC 9015

Extensive efforts to identify the diphtheria toxin in culture filtrates of C. ulcerans strain ATCC 9015 was unsuccessful. The culture filtrates did contain a component (called D-antigen) which formed a precipitin line with commercial diphtheria antitoxin. This component was purified by immunosorbent techniques using immobilized preparations of diphtheria antitoxin. It was shown, that D-antigen has a molecular weight of 84000, is not toxic for guinea-pigs, and has nothing in common with purified diphtheria toxin when tested in gel-diffusion tests. The lack of diphtheria toxin in form of its fragments in culture filtrates concentrated 20-fold was proved by the highly sensitive and specific viroimmunoassay.

Purification and some properties of exotoxin from Corynebacterium ulcerans strain ATCC 9015

Corynebacterium ulcerans strain ATCC 9015 develops during growth in environmental media an exotoxin which is lethal for guinea-pigs. The exotoxin has been purified by ion-exchange chromatography on DEAE-Sephadex A-50, affinity chromatography using immunosorbent techniques, and gel-filtration through Sephadex G-75. The exotoxin is of a protein nature and consists perhaps of two subunits. The molecular weights of the exotoxin and its subunits as estimated by SDS-PAGE were 40 000, 25 000, and 15 000, respectively. The toxic properties of exotoxin were destroyed upon heating (60 degrees C, 30 min), treatment with trypsin (10 micrograms/ml, 37 degrees C, 30 min), and by the extremes of pH (pH 4.0 or 10;0 24 hrs). Homologous antibodies readily neutralized the exotoxin while commercial diphtheria antitoxin had no effects. No immunological cross-reactivity was revealed between exotoxin of C. ulcerans and other bacterial toxins such as diphtheria toxin, enterotoxin from V. cholerae, and exotoxin A of P. aeruginosa.