Description of Mycobacterium pinniadriaticum sp. nov., isolated from a noble pen shell (Pinna nobilis) population in Croatia

Silvio Špičić; Sanja Duvnjak; Bojan Papić; Irena Reil; Snježana Zrnčić; Željko Mihaljević; Šimun Naletilić; Ivana Giovanna Zupičić; Gordan Kompes; Boris Habrun; Ivana Mareković; Maja Zdelar-Tuk

doi:10.3389/fmicb.2023.1289182

Description of Mycobacterium pinniadriaticum sp. nov., isolated from a noble pen shell (Pinna nobilis) population in Croatia

Front Microbiol. 2023 Dec 15:14:1289182. doi: 10.3389/fmicb.2023.1289182. eCollection 2023.

Authors

Affiliations

¹ Laboratory for Bacterial Zoonosis and Molecular Diagnostics of Bacterial Diseases, Department of Bacteriology and Parasitology, Croatian Veterinary Institute, Zagreb, Croatia.
² Veterinary Faculty, Institute of Microbiology and Parasitology, University of Ljubljana, Ljubljana, Slovenia.
³ Laboratory for Fish Pathology, Department for Pathological Morphology, Croatian Veterinary Institute, Zagreb, Croatia.
⁴ Laboratory for Pathology, Department for Pathological Morphology, Croatian Veterinary Institute, Zagreb, Croatia.
⁵ Laboratory for General Bacteriology and Mycology, Department of Bacteriology and Parasitology, Croatian Veterinary Institute, Zagreb, Croatia.
⁶ Department of Clinical and Molecular Microbiology, University Hospital Centre Zagreb, Zagreb, Croatia.

^# Contributed equally.

Abstract

Introduction: Shortly before the mass mortality event of the noble pen shell (Pinna nobilis) population in the south-eastern Adriatic coast, two rapidly growing Mycobacterium strains CVI_P3^T (DSM 114013 T, ATCC TSD-295 T) and CVI_P4 were obtained from the organs of individual mollusks during the regular health status monitoring.

Methods: The strains were identified as members of the genus Mycobacterium using basic phenotypic characteristics, genus-specific PCR assays targeting the hsp65 and 16S rRNA genes and the commercial hybridization kit GenoType Mycobacterium CM (Hain Lifescience, Germany). MALDI-TOF mass spectrometry did not provide reliable identification using the Bruker Biotyper Database.

Results and discussion: Genome-wide phylogeny and average nucleotide identity (ANI) values confirmed that the studied strains are clearly differentiated from their closest phylogenetic relative Mycobacterium aromaticivorans and other validly published Mycobacterium species (ANI ≤ 85.0%). The type strain CVI_P3^T was further characterized by a polyphasic approach using both phenotypic and genotypic methods. Based on the phenotypic, chemotaxonomic and phylogenetic results, we conclude that strains CVI_P3^T and CVI_P4 represent a novel species, for which the name Mycobacterium pinniadriaticum sp. nov. is proposed.

Keywords: Adriatic Sea; Croatia; Mycobacterium; average nucleotide identity; mass mortality event; noble pen shell.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was technically and financially supported by the Croatian Veterinary Institute, Zagreb, Croatia.