In vitro hemocyte phagocytosis activation after experimental infection of common octopus, Octopus vulgaris (Cuvier, 1797) with Photobacterium damselae subsp. piscicida or Vibrio alginolyticus at different temperatures and infection routes

Abstract

Due to the fast growth rate, the short life cycle, the high market price and the high food conversion efficiency, O. vulgaris is considered as a good candidate for aquaculture. One of the prerequisites for the successful integration of new species, such as octopi, into industrial-scale production, is the knowledge of the pathological conditions that may arise, with emphasis on infectious diseases caused by microorganisms and para-sites transmitted through wild populations, especially for the farmed organisms cul-tured in cages in proximity to teleost fish. The main objectives of this study were to investigate the sensitivity of common octopus to experimental infection with pathogenic bacteria, to assess the activation of hemocytes and more specifically their phagocytic activity after infection and to associate sensitivity of the species and phagocytic activity of hemocytes to temperature changes, route of infection and pathogen. Common octopus individuals were intramuscularly and intravenously infected with either Photobacterium damselae subsp. piscicida or Vibrio alginolyticus. The hemocyte phagocytosis activation in vitro at two temperatures (21 ± 0.5 °C and 24 ± 0.5 °C) was studied, in an effort to relate these aspects to climate change. Hemolymph was withdrawn on days 0, 3 and 7 post infections/injections. Number of circulating hemocytes/ml hemolymph, phagocytosis ability and Phagocytosis Particle Binding Intensity index were determined. Correlations between hemocytes and bodyweight and between hemocytes and phagocytosis ability were also determined. No mortalities were recorded irrespective of pathogen, route of infection and temperature employed. Circulating hemocytes in control specimens ranged between 1.60x10⁵ hemocytes ml^-1 hemolymph to 20.02x10⁵ hemocytes/ml hemolymph at both experiments and temperatures. The interrelation between octopi weight and circulating hemocytes showed that natural fluctuations, age, maturity stage and temperature may affect this relationship. Rise of temperature influenced phagocytosis which seemed to be route of infection, time-point and pathogen related. Specimens infected with Photobacterium damselae subsp. piscicida showed decreased phagocytosis with rise of temperature while when Vibrio alginolyticus was used, phagocytosis activity increased, in most cases. Temperature also played an important role in the correlation between the circulating hemocytes and phagocytosis activity, as at lower temperatures a negative strong correlation was observed. The results prompted us to calculate the activation index. This index showed that temperature is an important factor in hemocyte activation since for Photobacterium damselae subsp. piscicida infected specimens, hemocytes were more activated at 21 ± 0.5℃ instead of 24 ± 0.5℃, and the opposite observed for Vibrio alginolyticus samples and only later post-infection. Comparing the phagocytosis ability results with those obtained from Particle Binding Intensity index important differences concerned mainly confidence levels. The use of phagocytosis ability instead of PBI index provides more accurate results.

Keywords: Hemocyte activation; Octopus vulgaris; Phagocytosis; Photobacterium damselae subsp. piscicida; Vibrio alginolyticus.