Establishment and Characterization of a Novel Gill Cell Line, LG-1, from Atlantic Lumpfish (Cyclopterus lumpus L.)

The use of lumpfish (Cyclopterus lumpus) as a cleaner fish to fight sea lice infestation in farmed Atlantic salmon has become increasingly common. Still, tools to increase our knowledge about lumpfish biology are lacking. Here, we successfully established and characterized the first Lumpfish Gill cell line (LG-1). LG-1 are adherent, homogenous and have a flat, stretched-out and almost transparent appearance. Transmission electron microscopy revealed cellular protrusions and desmosome-like structures that, together with their ability to generate a transcellular epithelial/endothelial resistance, suggest an epithelial or endothelial cell type. Furthermore, the cells exert Cytochrome P450 1A activity. LG-1 supported the propagation of several viruses that may lead to severe infectious diseases with high mortalities in fish farming, including viral hemorrhagic septicemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV). Altogether, our data indicate that the LG-1 cell line originates from an epithelial or endothelial cell type and will be a valuable in vitro research tool to study gill cell function as well as host-pathogen interactions in lumpfish.

Effects of chlorpyrifos on primary gill cell culture of Lake Van fish (Alburnus tarichi Güldenstaadt 1814)

Lake Van fish (Alburnus tarichi Guldenstadt 1814) is the only fish that can adapt to the extreme conditions (pH 9.8 salinity 0.2% and alkalinity 151.2 meq/L) of Lake Van. In this study, it was aimed to determine the cytotoxic and genotoxic effects of chlorpyrifos (CPF) on Lake Van fish primary gill cell culture. Gill epithelium from Lake Van fish was isolated enzymatically and grown in primary culture on Leibovitz’s L-15 medium. After different doses (0.01, 0.1, 1, and 10 μM) of CPF were applied to the gill cells, the total oxidant status (TOS), total antioxidant status (TAS), malondialdehyde (MDA), and DNA damage levels (8-hydroxyguanine (8-OHdG)) were examined at the end of 24 and 48 h. It was determined that the TOS, MDA, and 8-OHdG levels increased in the cells exposed to high doses (1 and 10 μM) of CPF and the TAS was decreased (P < 0.05). It was revealed from this study that CPF administered at a dose higher than 1 μM can cause oxidative stress and DNA damage in the primary gill cell culture of Lake Van fish. In addition, the findings showed that Lake Van fish primary gill cell culture was useful in determining the effects of toxic substances likely to be the contaminants of a lake.

Screening of Chilean fish-killing microalgae using a gill cell-based assay

Fish-killing algal species are responsible for important global economic losses to the finfish industry. Chile is the world’s second-largest salmon exporter, and fish-killing algal blooms have widely impacted its production in the last decades. The lack of standardized analytical methods to quantify and characterize the so-called “ichthyotoxins” has hindered our understanding of the underlying ichthyotoxic modes of action. The novel application of a highly sensitive and reproducible fish RTgill-W1 cell line-based assay has allowed significant progress in the field. In this study, the ichthyotoxic potency of the main microalgae species, which has been reported in fish-killing events in the historical Chilean monitoring programs, was assessed. The dinoflagellate Karenia selliformis was the most ichthyotoxic species against the RTgill-W1 (cell viability down to 8%), representing the major threat for the local salmon industry. In comparison, the raphidophyte Heterosigma akashiwo and the dinoflagellate Prorocentrum micans were the least toxic (gill cell viability down to 81 and 89%, respectively). Importantly, ichthyotoxic flagellates were more toxic to fish gill cells upon rupture. These results have important implications for the mitigation and management of algal blooms by the salmon industry.