# Copper Pyrithione Induces Hepatopancreatic Apoptosis and Metabolic Disruption in Litopenaeus vannamei: Integrated Transcriptomic, Metabolomic, and Histopathological Analysis

## TL;DR

Copper pyrithione harms shrimp by causing liver-like organ damage, disrupting metabolism, and weakening immunity, according to a multi-omics study.

## Contribution

This study is the first to integrate transcriptomic, metabolomic, and histopathological data to reveal CuPT's toxicity mechanisms in marine crustaceans.

## Key findings

- CuPT exposure caused hepatopancreatic apoptosis and structural damage in shrimp.
- CuPT disrupted energy and fatty acid metabolism, leading to reactive oxygen species accumulation.
- Immune-related genes were downregulated, impairing detoxification and pathogen defense.

## Abstract

Copper pyrithione (CuPT), an antifoulant used in ship coatings, accumulates in coastal sediments and poses ecotoxicological risks to benthic organisms. This study investigated the molecular response mechanisms in the hepatopancreas of Litopenaeus vannamei exposed to CuPT (128 μg/L) for 3 and 48 h. Apoptosis detection, transcriptomic, and metabolomic analysis indicated that prolonged exposure induced hepatopancreatic, apoptosis, impaired energy, and fatty acid metabolism. Concurrently, down-regulation of immune-related genes compromised detoxification and pathogen defense mechanisms, threatening their health and growth. These findings highlight the ecological risks of CuPT to marine crustaceans and provide critical insights into its toxicity mechanisms, offering scientific references for regulating antifouling biocides.

Copper pyrithione (CuPT), an emerging biocide used in ship antifouling coatings, may accumulate in marine sediments and pose risks to non-target organisms. However, current research on CuPT toxicity remains limited. Litopenaeus vannamei, one of the world’s most important aquaculture shrimp species, relies heavily on its hepatopancreas for energy metabolism, detoxification, and immune responses. Due to their benthic habitat, these shrimps are highly vulnerable to contamination in sediment environments. This study investigated the toxicological response in the hepatopancreas of L. vannamei exposed to CuPT (128 μg/L) for 3 and 48 h. Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL) fluorescence staining revealed increased apoptosis, deformation of hepatic tubule lumens, and the loss of stellate structures in the hepatopancreas after CuPT 48 h exposure. A large number of differentially expressed genes (DEGs) were identified by transcriptomics analysis at 3 and 48 h, respectively. Most of these DEGs were related to detoxification, glucose transport, and immunity. Metabolomic analysis identified numerous significantly different metabolites (SDMs) at both 3 and 48 h post-exposure, with most SDMs associated with energy metabolism, fatty acid metabolism, and related pathways. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of metabolomics and transcriptome revealed that both DEGs and SDMs were enriched in arachidonic acid metabolism, fatty acid biosynthesis, and glycolysis/gluconeogenesis pathways at 3 h, while at 48 h they were enriched in the starch and sucrose metabolism, amino sugar and nucleotide sugar metabolism, and galactose metabolism pathways. These results suggested that CuPT disrupts the energy and lipid homeostasis of L. vannamei. This disruption compelled L. vannamei to allocate additional energy toward sustaining basal physiological functions and consequently caused the accumulation of large amounts of reactive oxygen species (ROS) in the body, leading to apoptosis and subsequent tissue damage, and ultimately suppressed the immune system and impaired the health of L. vannamei. Our study elucidates the molecular mechanisms of CuPT-induced metabolic disruption and immunotoxicity in L. vannamei through integrated multi-omics analyses, providing new insights for ecological risk assessment of this emerging antifoulant.