Effects of Zinc Oxide Nanoparticles (ZnO NPs) on Growth, Immune Responses and Histopathological Alterations in Asian Seabass (Lates calcarifer, Bloch 1790) under Low-Salinity Conditions

In the present study, Asian seabass (<i>Lates calcarifer</i>, Bloch) fingerings were used as an animal model to investigate the toxicological effects of zinc oxide nanoparticles (ZnO NPs) under 5 ppt estuarine conditions. The fish were exposed to 0, 1, 5 or 50 ppm ZnO NPs for 8 weeks. It was found that ZnO NP concentrations of 5-50 ppm negatively affected several growth rate parameters, such as the weight and total length of the fish. Additionally, 5 and 50 ppm ZnO NPs led to 32.55% and 100% mortality, respectively, after 8 weeks after exposure (WAE). Furthermore, compared with the control, exposure to 1-50 ppm ZnO NPs strongly affected hematological indices, such as total blood cells, red blood cells, leukocytes and hematocrit, and suppressed lysozyme activity, superoxide anion production and bactericidal activity. High Zn concentrations accumulated in the head kidney, gills and liver, whereas low levels were detected in the gut, skin and muscle. Expression analysis of immune-related genes via quantitative real-time RT-PCR revealed that 5 and 50 ppm ZnO NPs significantly upregulated the <i>cc</i> and <i>cd4</i> genes at 1 WAE. In contrast, 50 ppm ZnNPs downregulated the expression levels of the <i>cd8</i>, <i>cc</i>, <i>hsp70</i>, <i>hsp90</i>, <i>tcrα</i>, <i>lyz</i> and <i>igmh</i> genes at 1 WAE (<i>p</i> < 0.05). Finally, at 8 WAE, histopathological analysis revealed that 5 and 50 ppm ZnO NPs severely induced alterations in the head kidney, gills and liver.