The early stages of the immune response of the European abalone Haliotis tuberculata to a Vibrio harveyi infection.

Vibrio harveyi is a marine bacterial pathogen responsible for episodic abalone mortalities in France, Japan and Australia. In the European abalone, V. harveyi invades the circulatory system in a few hours after exposure and is lethal after 2 days of infection. In this study, we investigated the responses of European abalone immune cells over the first 24 h of infection. Results revealed an initial induction of immune gene expression including Rel/NF-kB, Mpeg and Clathrin. It is rapidly followed by a significant immuno-suppression characterized by reduced cellular hemocyte parameters, immune response gene expressions and enzymatic activities. Interestingly, Ferritin was overexpressed after 24 h of infection suggesting that abalone attempt to counter V. harveyi infection using soluble effectors. Immune function alteration was positively correlated with V. harveyi concentration. This study provides the evidence that V. harveyi has a hemolytic activity and an immuno-suppressive effect in the European abalone.

Vibrio harveyi adheres to and penetrates tissues of the European abalone Haliotis tuberculata within the first hours of contact.

Vibrio harveyi is a marine bacterial pathogen responsible for episodic epidemics generally associated with massive mortalities in many marine organisms, including the European abalone Haliotis tuberculata. The aim of this study was to identify the portal of entry and the dynamics of infection of V. harveyi in the European abalone. The results indicate that the duration of contact between V. harveyi and the European abalone influences the mortality rate and precocity. Immediately after contact, the epithelial and mucosal area situated between the gills and the hypobranchial gland was colonized by V. harveyi. Real-time PCR analyses and culture quantification of a green fluorescent protein-tagged strain of V. harveyi in abalone tissues revealed a high density of bacteria adhering to and then penetrating the whole gill-hypobranchial gland tissue after 1 h of contact. V. harveyi was also detected in the hemolymph of a significant number of European abalones after 3 h of contact. In conclusion, this article shows that a TaqMan real-time PCR assay is a powerful and useful technique for the detection of a marine pathogen such as V. harveyi in mollusk tissue and for the study of its infection dynamics. Thus, we have revealed that the adhesion and then the penetration of V. harveyi in European abalone organs begin in the first hours of contact. We also hypothesize that the portal of entry of V. harveyi in the European abalone is the area situated between the gills and the hypobranchial gland.

The impacts of handling and air exposure on immune parameters, gene expression, and susceptibility to vibriosis of European abalone Haliotis tuberculata.

Wild or farmed abalone are regularly exposed to stressors, such as air exposure and handling. Immune and transcriptional responses as well as susceptibility to vibriosis of sexually mature or immature European abalone acclimated at 16 or 19 °C were determined following handling or air exposure. Hemocyte density and H2O2 production increased while hemocyte viability and phagocytic index decreased following handling. Air exposure induces a decrease of hemocyte density and phagocytic index. Measurement of the expression of genes implicated in general metabolic, immunological and stress responses in gills, foot-muscle and hemocytes by real time q-PCR suggested that both stressors lead to a metabolic rate depression, characterized by a general inhibition of transcription. Finally, following handling a Vibrio harveyi challenge enhances almost 100% mortality of sexually immature animals at 19 °C while it has been previously demonstrated that only mature are susceptible to vibriosis.

Gene expression patterns of abalone, Haliotis tuberculata, during successive infections by the pathogen Vibrio harveyi.

Since 1998, episodic mass mortality of the abalone Haliotistuberculata has been observed along the northern Brittany coast of France caused by a complex interaction among the host, pathogen and environmental factors. In the present study, abalone were submitted to two successive infections with the pathogen Vibrioharveyi under controlled conditions. During the first challenge, infection by V.harveyi resulted in 64% mortality of mature abalone. After a second infection of those surviving the first challenge, only 44% mortality was observed. Physiological variability in the host response appears to be a major determinant in susceptibility to V.harveyi. In order to isolate differentially expressed genes in H.tuberculata challenged with this bacterium, suppression subtractive hybridization (SSH) cDNA libraries were constructed from muscle of moribund abalone (susceptibles), surviving individuals (apparently resistant to the bacterium) and control (unexposed) animals. Of the 1152 clones sequenced, 218 different partial cDNA sequences were obtained and represented 69 known genes. Of these, 65 were identified for the first time in H.tuberculata. Using real-time PCR, a time-course study was conducted on 19 of the genes identified by SSH. A majority of differentially expressed transcripts were down-regulated in susceptible individuals as compared to their resistant counterparts. Bacterial challenge of abalone resulted in the up-regulation of three transcripts (encoding ferritin, heat shock protein HSP84 and fatty acid binding protein FABP) in those that survived exposure to V.harveyi. This study has identified potential candidates for further investigation into the functional basis of resistance and susceptibility to summer vibriosis outbreaks in abalone.