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Sci Rep
2022 Mar 22;121:4859. doi: 10.1038/s41598-022-08772-2.
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Evidence for association of Vibrio echinoideorum with tissue necrosis on test of the green sea urchin Strongylocentrotus droebachiensis.
Hira J
,
Stensvåg K
.
Abstract
"Sea urchin lesion syndrome" is known as sea urchin disease with the progressive development of necrotic epidermal tissue and loss of external organs, including appendages on the outer body surface. Recently, a novel strain, Vibrio echinoideorum has been isolated from the lesion of green sea urchin (Strongylocentrotus droebachiensis), an economically important mariculture species in Norway. V. echinoideorum has not been reported elsewhere in association with green sea urchin lesion syndrome. Therefore, in this study, an immersion based bacterial challenge experiment was performed to expose sea urchins (wounded and non-wounded) to V. echinoideorum, thereby mimicking a nearly natural host-pathogen interaction under controlled conditions. This infection experiment demonstrated that only the injured sea urchins developed the lesion to a significant degree when exposed to V. echinoideorum. Pure cultures of the employed bacterial strain were recovered from the infected animals and its identity was confirmed by the MALDI-TOF MS spectra profiling. Additionally, the hemolytic phenotype of V. echinoideorum substantiated its virulence potential towards the host, and this was also supported by the cytolytic effect on red spherule cells of sea urchin. Furthermore, the genome sequence of V. echinoideorum was assumed to encode potential virulence genes and were subjected to in silico comparison with the established virulence factors of Vibrio vulnificus and Vibrio tasmaniensis. This comparative virulence profile provided novel insights about virulence genes and their putative functions related to chemotaxis, adherence, invasion, evasion of the host immune system, and damage of host tissue and cells. Thus, it supports the pathogenicity of V. echinoideorum. In conclusion, the interaction of V. echinoideorum with injured sea urchin facilitates the development of lesion syndrome and therefore, revealing its potentiality as an opportunistic pathogen.
Figure 1. Lesion syndrome of green sea urchin (S. droebachiensis) reported in Northern Norway. The animal here is presented up-side down with mouth located on the top.
Figure 2. Green sea urchins (S. droebachiensis) before (a, b) and after (c, d) the in vivo bacterial challenge experiment with V. echinoideorum at ± 6–8 °C. (a) Healthy sea urchin before the challenge experiment and a closer macroscopic view of intact spines, pedicellariae, tube feet, and other external appendages. (b) Artificially wounded sea urchin (spines, pedicellariae, tube feet, and other external appendages were trimmed). (c) Recovery of spines, pedicellariae, tube feet, and other external appendages of artificially wounded sea urchin (control, not exposed to bacteria) at the end of the challenge experiment. (d) Artificially wounded sea urchin (exposed to bacteria) having necrotized tissues, devoid of spines, pedicellariae, tube feet, and other external appendages.
Figure 3. Internal view of green sea urchins (S. droebachiensis) hard body shells (test) after the in vivo bacterial challenge experiment with V. echinoideorum at ± 6–8 °C. (a) Healthy interior of the ambulacral and interambulacral test zone. (b) and (c) represents the intact ambulacral and interambulacral area of a healthy sea urchins. Here, (d) and (e) depicts the artificially wounded sea urchins without exposure to bacteria. No signs of calcareous skeleton, blackish patches, swelling of tube feet pores and darkened perforations are seen, whereas (f) and (g) represents the artificially wounded, bacterial exposed sea urchins with signs of skeletal damage. Scale bar 2 mm.
Figure 4. Cytolytic properties of V. echinoideorum towards sheep RBCs and sea urchin RSCs. (a) Hemolysis reaction against sheep RBCs is presented here with a clear zone around the colony (16 mm in diameter zone) on blood agar. (b) Control with RSCs not exposed to bacteria, (c) RSCs exposed to bacteria. Scale bar 60 µm.
Figure 5. A circos plot featuring the proteome comparison of predicted virulence genes of V. echinoideorum. From the outer ring to the inner ring: V. echinoideorum, V. tasmaniensis LGP32, V. vulnificus YJ016, and V. echinoideorum (as control replicate). Only distinguished features between these strains are labelled.
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