16S rRNA Gene Sequencing, Multilocus Sequence Analysis, and Mass Spectrometry Identification of the Proposed New Species “Clostridium neonatale”

Abstract

In 2002, an outbreak of necrotizing enterocolitis in a Canadian neonatal intensive care unit was associated with a proposed novel species of Clostridium, "Clostridium neonatale." To date, there are no data about the isolation, identification, or clinical significance of this species. Additionally, C. neonatale has not been formally classified as a new species, rendering its identification challenging. Indeed, the C. neonatale 16S rRNA gene sequence shows high similarity to another Clostridium species involved in neonatal necrotizing enterocolitis, Clostridium butyricum. By performing a polyphasic study combining phylogenetic analysis (16S rRNA gene sequencing and multilocus sequence analysis) and phenotypic characterization with mass spectrometry, we demonstrated that C. neonatale is a new species within the Clostridium genus sensu stricto, for which we propose the name Clostridium neonatale sp. nov. Now that the status of C. neonatale has been clarified, matrix-assisted laser desorption ionizationtime of flight mass spectrometry (MALDI-TOF MS) can be used for better differential identification of C. neonatale and C. butyricum clinical isolates. This is necessary to precisely define the role and clinical significance of C. neonatale, a species that may have been misidentified and underrepresented during previous neonatal necrotizing enterocolitis studies.

Description of Paraclostridium bifermentans subsp. muricolitidis subsp. nov., emended description of Paraclostridium bifermentans (Sasi Jyothsna et al., 2016), and creation of Paraclostridium bifermentans subsp. bifermentans subsp. nov

“…MALDI‐TOF MS is used as a classification tool in bacterial taxonomy of genus Clostridium species . MALDI‐TOF MS and cluster analysis was performed on the three strains analyzed here.…”

16S rRNA Gene sequencing: A practical approach to confirming the identity of food borne bacteria

“…Such approaches are time-consuming often considerably more in the case of anaerobes and other fastidious organisms such as Clostridium species. Hence conventional method of identification is rigorous and frequently fails to produce relevant data within the crucial timeline to permit rapid and appropriate therapeutic decisions to be made (Bouvet et al, 2014;Rossi-Tamisier et al, 2015). In view of the fact that many diverse species can share the same biochemical phenotypes; there has been debate on whether conventional approaches can correctly distinguish between isolates (Srinivasan et al, 2015).…”

Microbial culturomics to isolate halophilic bacteria from table salt: genome sequence and description of the moderately halophilic bacterium Bacillus salis sp. nov.

“…The emergence of new tools for DNA sequencing and technology, such as matrix-assisted desorption ionization–time of flight mass spectrometry (MALDI-TOF MS), has allowed an increase in available genomic and proteomic data over the last few years [8] , [9] . These technologic advances have allowed us to develop a new way of describing bacterial species that takes into account genomic and protonic information [10] .…”