FN Archimer Export Format
PT J
TI Characterization of abalone Haliotis tuberculata-Vibrio harveyi interactions in gill primary cultures
BT 
AF PICHON, Delphine
   CUDENNEC, Benoit
   HUCHETTE, Sylvain
   DJEDIAT, Chakib
   RENAULT, Tristan
   PAILLARD, Christine
   AUZOUX-BORDENAVE, Stephanie
AS 1:1;2:2,3;3:4;4:5;5:3;6:6;7:1,7;
FF 1:;2:;3:;4:;5:PDG-RBE-SG2M;6:;7:;
C1 Museum Natl Hist Nat, DMPA, UMR BOREA 7208 CNRS MNHN IRD UPMC, Stn Biol Marine, F-29900 Concarneau, France.
   Univ Lille 1, Lab ProBioGEM, F-59655 Villeneuve Dascq, France.
   IFREMER, Lab Genet & Pathol Mollusques Marins, Unite Sante Genet & Microbiol Mollusques, F-17390 La Tremblade, France.
   France Haliotis, F-29880 Plouguerneau, France.
   Museum Natl Hist Nat, F-75005 Paris, France.
   LEMAR Univ Bretagne Occidentale, CNRS UBO Ifremer IRD UMR6539, F-29280 Plouzane, France.
   Univ Paris 06, F-75005 Paris, France.
C2 MNHN, FRANCE
   UNIV LILLE, FRANCE
   IFREMER, FRANCE
   FRANCE HALIOTIS, FRANCE
   MNHN, FRANCE
   UBO, FRANCE
   UNIV PARIS 06, FRANCE
SI LA TREMBLADE
   BREST
SE PDG-RBE-SG2M
   UBOLEMAR
IN WOS Ifremer jusqu'en 2018
   copubli-france
   copubli-univ-france
IF 1.449
TC 14
UR https://archimer.ifremer.fr/doc/00146/25683/23745.pdf
LA English
DT Article
DE ;Haliotis tuberculata;Vibrio harveyi;Gill cell culture;Pathogenicity;Phenoloxidase;Phagocytosis
AB The decline of European abalone Haliotis tuberculata populations has been associated with various pathogens including bacteria of the genus Vibrio. Following the summer mortality outbreaks reported in France between 1998 and 2000, Vibrio harveyi strains were isolated from moribund abalones, allowing in vivo and in vitro studies on the interactions between abalone H. tuberculata and V. harveyi. This work reports the development of primary cell cultures from abalone gill tissue, a target tissue for bacterial colonisation, and their use for in vitro study of host cell—V. harveyi interactions. Gill cells originated from four-day-old explant primary cultures were successfully sub-cultured in multi-well plates and maintained in vitro for up to 24 days. Cytological parameters, cell morphology and viability were monitored over time using flow cytometry analysis and semi-quantitative assay (XTT). Then, gill cell cultures were used to investigate in vitro the interactions with V. harveyi. The effects of two bacterial strains were evaluated on gill cells: a pathogenic bacterial strain ORM4 which is responsible for abalone mortalities and LMG7890 which is a nonpathogenic strain. Cellular responses of gill cells exposed to increasing concentrations of bacteria were evaluated by measuring mitochondrial activity (XTT assay) and phenoloxidase activity, an enzyme which is strongly involved in immune response. The ability of gill cells to phagocyte GFP-tagged V. harveyi was evaluated by flow cytometry and gill cells-V. harveyi interactions were characterized using fluorescence microscopy and transmission electron microscopy During phagocytosis process we evidenced that V. harveyi bacteria induced significant changes in gill cells metabolism and immune response. Together, the results showed that primary cell cultures from abalone gills are suitable for in vitro study of host-pathogen interactions, providing complementary assays to in vivo experiments.
PY 2013
PD OCT
SO Cytotechnology
SN 0920-9069
PU Springer
VL 65
IS 5
UT 000326053500041
BP 759
EP 772
DI 10.1007/s10616-013-9583-1
ID 25683
ER

EF