Fig. 3

Anisakis pegreffii L2 hatched in vitro and cultured up to 24 h (a, b), 48 h (c), 72 h (d), 1 week (e–h) and 2 weeks (i, j) in Schneider’s Drosophila media, and 2 weeks in seawater (control; k, l). DAPI (blue) is localising larval nuclei. Insets show details under higher magnification: (c) no large-nucleus cell attributable to the excretory gland cell is observable; d the first appearance of a larger-nucleus cell; and e-i presumable excretory gland cell. Note the increase in DAPI intensity in this cell through larval growth. White and black arrows indicate presumable excretory gland cell in fluorescence and bright-field pictures, respectively. Fluorescence is overlaid on the complementary bright field for better orientation (a and b; e and f; g and h; i and j; k and l). Scale bars: 10 µm (g, h, and insets in c, d and i); 20 µm (the rest)