Establishment and partial characterization of a cell line from burbot Lota lota maculosa: susceptibility to IHNV, IPNV and VHSV

Abstract

This study describes the development and partial characterization of a continuous fibroblastic-like cell line (BEF-1) developed from late stage embryos of North American burbot Lota lota maculosa. This cell line has been maintained for over 5 yr and 100 passages in vitro. Cells were cultured using Eagle's minimum essential medium with Earle's salts (MEM) supplemented with GlutaMAX™, and 10% fetal bovine serum (FBS), pH 7.4. The addition of penicillin-streptomycinneomycin (PSN) antibiotic mixture (0.05, 0.05, 0.1 mg ml -1 , respectively) did not negatively influence cell replication; however, the antimycotic Fungizone™ (2.5 µg ml -1 , amphotericin B) caused cell rounding and resulted in a severe decrease in cell proliferation. Optimal incubation temperature has been observed between 15 and 23°C, and at these temperatures cultures are routinely passed using standard trypsinization methods every 5 to 7 d at a split ratio of 1:3 or 1:4. The cell line was susceptible to isolates of the M and U North American genotypes of infectious hematopoietic necrosis virus (IHNV), and to isolates of genotypes I, IVa, and IVb of viral hemorrhagic septicemia virus (VHSV). In contrast, the cell line was refractory to infection by 2 North American isolates of infectious pancreatic necrosis virus (IPNV) from serotypes A1 and A9. This cell line provides a new laboratory tool, will allow further investigation into viral diseases of burbot and possibly other species, and is the first immortalized cell line reported from a species in the Gadidae (cod) family.

Performance and Macronutrient Composition of Age‐0 Burbot Fed Four Diet Treatments

“…Beginning in 2003, an experimental research program initiated at the University of Idaho's Aquaculture Research Institute (UIARI) focused on assessing the feasibility of developing burbot culture as a population restoration measure. Spawning, sperm motility inhibition, and semen cryopreservation methods were developed first (Zuccarelli et al 2007;Jensen et al 2008aJensen et al , 2008b, followed by incubation methods and larval and juvenile rearing strategies (Jensen et al 2008c;Polinski et al 2010c), and then burbot disease susceptibility was characterized (Polinski et al 2010a(Polinski et al , 2010b. Collectively, these efforts enabled the first releases of cultured burbot of ages 0, 1, 2, and 3 years for conservation purposes in British Columbia and Idaho in 2009.…”