Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

Ruben Avendaño-Herrera, Eloisa Arias-Muñoz, Verónica Rojas, Alicia E. Toranzo, Matías Poblete-Morales, Claudio Córdova, Rute Irgang

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Vibrio ordalii is an extracellular, Gram-negative bacterium that produces vibriosis in salmonids. While pathogenesis is not fully understood, this bacterium has numerous likely genes for adhesion, colonization, invasion factors and, as recently suggested, intracellular behaviour. Therefore, this study aimed to clarify possible intracellular behaviour for V. ordalii Vo-LM-18 and ATCC 33509T in the fish-cell lines SHK-1 and CHSE-214. Confocal microscopy revealed Vo-LM-18 and ATCC 33509T inside cytoplasm in both fish-cell lines at 4 hr post-inoculation (hpi). At 8 and 16 hpi, the proportion of fish cells invaded by both strains increased. Moreover, intracellular V. ordalii were observed after 8 hpi inside mouse embryonic fibroblasts (MEF), demonstrating that entry was not due to a cellular phagocytosis process. Flow cytometry confirmed immunocytochemistry results, with both V. ordalii evidencing statistically significant differences in the number of infected cells between 8 and 16 hpi. Interestingly, V. ordalii infection did not significantly damage fish cells, as determined by LDH liberation. Viable counts at 8 hpi detected, on average for both lines, 176 ± 47 CFU/ml of culturable intracellular Vo-LM-18 and ATCC 33509T cells. These in vitro findings support the facultative intracellular behaviour of V. ordalii and may be of importance for understanding pathogenicity and survival in aquatic environments.

Original languageEnglish
Pages (from-to)1447-1455
Number of pages9
JournalJournal of Fish Diseases
Volume42
Issue number10
DOIs
StatePublished - 1 Oct 2019
Externally publishedYes

Keywords

  • Vibriosis
  • intracellular facultative
  • virulence mechanism

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