Resistance of the oyster pathogen Vibrio tasmaniensis LGP32 against grazing by Vannella sp . marine amoeba involves Vsm and CopA virulence factors

ibrios are ubiquitous in marine environments and opportunistically colonize a broad range of hosts. Strains of Vibrio tasmaniensis present in oyster farms can thrive in oysters during juvenile mortality events and behave as facultative intracellular pathogen of oyster hemocytes. Herein, we wondered whether V. tasmaniensis LGP32 resistance to phagocytosis is specific to oyster immune cells or contributes to resistance to other phagocytes, like marine amoebae. To address this question, we developed an integrative study, from the first description of amoeba diversity in oyster‐farms to the characterization of LGP32 interactions with amoebae. An isolate of the Vannella genus, Vannella sp. AP1411, which was collected from oyster farms, is ubiquitous, and belongs to one clade of Vannella that could be found associated with Vibrionaceae. LGP32 was shown to be resistant to grazing by Vannella sp. AP1411 and this phenotype depends on some previously identified virulence factors: secreted metalloprotease Vsm and copper efflux p‐ATPase CopA, which act at different steps during amoeba‐vibrio interactions, whereas some other virulence factors were not involved. Altogether, our work indicate that some virulence factors can by involved in multi‐host interactions of V. tasmaniensis ranging from protozoans to metazoans, potentially favoring their opportunistic behavior.

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Fig. S1 Phylogeny of the diversity of amoebae OTUs found by barcoding of the v7 region of the 18 s rDNA gene.
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Fig. S2 Vannella sp. AP1411 isolated from Thau lagoon, France.
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Fig. S3 Quantification of Vannella sp. AP1411 phagocytosis by cytometry. (A)
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Fig. S4 CusAB, SodA, Vsp, InhA, VipA1 and VipA2 are not involved in LGP32 resistance to grazing by Vannella sp. 1411.
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Fig. S5 Vsm or CopA do not affect the phagocytosis rate of LGP32 by the amoeba Vannella sp. AP1411
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Table S1 Sanger sequences of sampled marine amoebae
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Supplementary Movie 1 Time‐lapse microscopy movie of Vannella sp. AP1411 and LGP32wt lawn in co‐culture.
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Supplementary Movie 2 Time‐lapse microscopy of Vannella sp. AP1411 and LGP32 ΔcopA lawn in co‐culture.
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Supplementary Movie 3 Time‐lapse microscopy of Vannella sp. AP1411 and LGP32 Δvsm lawn in co‐culture.
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How to cite
Robino Etienne, Poirier Aurore C., Amraoui Hajar, Le Bissonnais Sandra, Perret Angelique, Lopez‐joven Carmen, Auguet Jean-Christophe, Rubio Tristan P., Cazevieille Chantal, Rolland Jean-Luc, Héchard Yann, Destoumieux‐garzón Delphine, Charrière Guillaume (2020). Resistance of the oyster pathogen Vibrio tasmaniensis LGP32 against grazing by Vannella sp . marine amoeba involves Vsm and CopA virulence factors. Environmental Microbiology. 22 (10). 4183-4197. https://doi.org/10.1111/1462-2920.14770, https://archimer.ifremer.fr/doc/00509/62109/

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