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. 2016 Oct 21:7:1656.
doi: 10.3389/fmicb.2016.01656. eCollection 2016.

A New Multilocus Sequence Typing Scheme and Its Application for the Characterization of Photobacterium damselae subsp. damselae Associated with Mortality in Cetaceans

Patricia Alba  1 , Andrea Caprioli  1 , Cristiano Cocumelli  2 , Angela Ianzano  1 , Valentina Donati  1 , Francesco Scholl  2 , Luigi Sorbara  1 , Giuliana Terracciano  3 , Gianluca Fichi  3 , Fabio Di Nocera  4 , Alessia Franco  1 , Antonio Battisti  1
Affiliations

A New Multilocus Sequence Typing Scheme and Its Application for the Characterization of Photobacterium damselae subsp. damselae Associated with Mortality in Cetaceans

Patricia Alba et al. Front Microbiol. .

Abstract

Photobacterium damselae subsp. damselae (PDD) is a known pathogen of fish, humans and marine mammals. In this study, a Multilocus Sequence Typing (MLST) scheme based on six housekeeping genes (glp, gyrB, metG, pnt, pyrC, and toxR) was developed to better understand the PDD population structure and used to type 73 PDD isolates from cetaceans, mainly striped dolphins (Stenella coeruleoalba) involved in mortality episodes, and from a few marine chelonians. Five reference ATCC strains were also included in the study. Typing allowed the discrimination of groups of PDD strains isolated from different host species, at different times and from different geographic areas, suggesting that a clonal PDD group may have spread in the Tyrrhenian sea at the time of an Unusual Mortality Event (UME) among cetaceans, mainly striped dolphins, occurred in early 2013 along the Italian western coasts.

Keywords: MLST; Photobacterium damselae; cetaceans; genotyping; infection; phylogeny; zoonoses.

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Figures

Figure 1
Figure 1
MST representation. Each node represents a different ST, which is indicated by a number. The number of partitions represents the number of isolates belonging to that ST. Bold continuous lines link isolates with STs which differ in one allele only. Light continuous lines link isolates with STs differing in two alleles. Dotted lines (bold or light) link isolates whose STs presented three or more alleles of difference. The shaded area includes the isolates belonging to the CC45. Yellow: PDD isolated before January 2013. Red: PDD isolated between January 2013 and March 2013. Orange: PDD isolated after March 2013. Blue: ATCC PDD and PDP strains.
Figure 2
Figure 2
Phylogenetic tree of the concatenated sequences built using Bionumerics. Phylogenetic tree constructed using Neighbor-joining method based on a multiple alignment. None position was discarded, having a total of 2739 bases in the final dataset. The tree topology was tested using 500 bootstrap. The haemolysin genes detected, the defined UME group, the CC, the ST, the haemolytic phenotype, the dates of isolation, the ID of the isolates, the sources of isolation and the hosts were included.
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