Detection and virulence potential of a phospholipase D-negative Corynebacterium ulcerans from a concurrent diphtheria and infectious mononucleosis case
Diphtheria by Corynebacterium ulcerans is increasingly occurring in children, adolescents and adults. In addition to diphtheria toxin (DT), phospholipase D (PLD) is considered a virulence factor of C. ulcerans. In the present study, a first case of concurrent diphtheria by a PLD-negative C. ulcerans and infectious mononucleosis (IM) was verified. Clinical and microbiological profiles and binding properties to human Fibrinogen (Fbg), Fibronectin (Fn) and type I collagen (col I) biotinylated proteins and virulence to Caenorhabditis elegans were investigated for C. ulcerans strain 2590 (clinical isolate) and two control strains, including PLD-positive BR-AD22 wild type and PLD-negative ELHA-1 PLD mutant strains. MALDI-TOF assays and a multiplex PCR of genes coding for potentially toxigenic corynebacteria identified strain 2590 as non-DT producing. Interestingly, strain 2590 did not express PLD activity in the CAMP test although the presence of the pld gene was verified. PLD-negative 2590 and a PLD-positive 210932 strains showed similar affinity to Fbg, Fn and type I collagen. C. elegans were able to escape from C. ulcerans strains, independent of PLD and DT production. Higher mortality of nematodes was verified for PLD-negative strains. Additional studies concerning multifactorial virulence potential of C. ulcerans, including environmental conditions remain necessary.
KeywordsCo-infection Corynebacterium ulcerans Diphtheria Infectious mononucleosis Virulence factors Zoonosis
This study was financied in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001 and Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Programa Estratégico de Apoio à Pesquisa em Saúde - Fundação Oswaldo Cruz (PAPES V-FIOCRUZ), Sub-Reitoria de Pós-graduação e Pesquisa da Universidade do Estado do Rio de Janeiro (SR-2/UERJ) and Friedrich-Alexander Universität-Erlangen-Nürnberg (FAU).
Conceived of or designed study: Liliane Simpson-Lourêdo, Maria Helena S. Villas Bôas, Ana Luíza Mattos-Guaraldi. Performed research: Liliane Simpson-Lourêdo, Cecília M.F. Silva, Nadjla F. Souza, Milena M. Santana. Analysed data: Liliane Simpson-Lourêdo, Raphael Hirata Jr, Andreas Burkovski, Maria Helena S. Villas Bôas, Ana Luíza Mattos-Guaraldi. Contributed new methods or models: Camila A. Antunes, Elena Hacker, Prescilla E. Nagao. Wrote the paper: Liliane Simpson-Lourêdo, Maria Helena S. Villas Bôas, Ana Luíza Mattos-Guaraldi.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The study (CAAE 44674314.3.3001.5091) was approved by the Research Ethics Committee of Hospital Universitário Pedro Ernesto/Universidade do Estado do Rio de Janeiro. The consent to participate was not required because the investigated isolate was taken as a part of standard care (diagnostic purposes).
- Clinical Laboratory Standards Institute (CLSI) (2015) Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolated or fastidious bacteria, approved guideline. CLSI document M45. ISBN 1-56238-917-3Google Scholar
- Haight K, Holden FA (1982) When infections co-exist: infectious mono and diphtheria. Can Fam Phys 28:785–788Google Scholar
- Konrad R, Berger A, Huber I et al (2010) Matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry as a tool for rapid diagnosis of potentially toxigenic Corynebacterium species in the laboratory management of diphtheria-associated bacteria. Eur Surveill 15(43):19699CrossRefGoogle Scholar
- Mattos-Guaraldi AL, Hirata R Jr, Azevedo VA (2013) Corynebacterium diphtheriae, Corynebacterium ulcerans and Corynebacterium pseudotuberculosis—general aspects. In: Burkovski A (ed) Corynebacterium diphtheriae and related toxigenic species. Springer, Erlangen, pp 15–27Google Scholar
- Perkins S, Cordery R, Nixon G et al (2010) Investigations and control measures following a non-travel-associated case of toxigenic Corynebacterium diphtheriae, London, United Kingdom, December 2009–January 2010. Euro Surveill 15:19544. https://doi.org/10.2807/ese.15.16.19544-en CrossRefPubMedGoogle Scholar
- World Health Organization (WHO) (2018) Diphtheria reported cases. http://apps.who.int/immunization_monitoring/globalsummary/timeseries/tsincidencediphtheria.html. Accessed 01 Sept 2018