A simple method to evaluate the toxic effects of Prorocentrum lima extracts to fish (sea bass) kidney cells.

ABSTRACT

The diarrhetic shellfish toxins (DSTs) okadaic acid (OA) and its analogues - the dinophysistoxins (DTXs) - are produced by dinoflagellates such as Prorocentrum lima and can bioaccumulate in filter-feeding organisms as they are transferred through the food web. Although there is no assessment of the harmful effects of these toxins on the fish's immune system, this study developed a primary culture protocol for kidney cells from marine fish Centropomus parallelus and evaluated the immunotoxic effects to P. lima extracts containing DSTs. The cells were obtained by mechanical dissociation, segregated with Percoll gradient, and incubated for 24 h at 28 °C in a Leibovitz culture medium supplemented with 2% fetal bovine serum and antibiotics. The exposed cells were evaluated in flow cytometry using the CD54 PE antibody. We obtained >5.0 × 106 viable cells per 1.0 g of tissue that exhibited no cell differentiation. Exposure to 1.2 or 12 ng DST mL-1 stimulated the immune system activation and increased the proportion of activated macrophages and monocytes in 48 to 52% and in 127 to 146%, respectively. The protocol proved to be an alternative tool to assess the immunotoxic effects of DST exposure on fish's anterior kidney cells.