Abstract
The spotted longbarbel catfish, Hemibagrus guttatus, a nationally protected Class II species in China, faces increasing threats from habitat degradation. Recently, the spotted longbarbel catfish has gained attention as a promising aquaculture species, not only for its premium flesh quality but also for its potential role in conservation through sustainable captive breeding programs. Ammonia nitrogen (ammonia-N) is a ubiquitous byproduct of intensive farming and serves as the primary environmental stressor confronting aquatic species. Elucidating the ammonia-N tolerance of spotted longbarbel catfish constitutes a critical prerequisite for its successful domestication, which is the aim of this study. We demonstrate that ammonia-N stress significantly decreases the survival rate of spotted longbarbel catfish and induces tissue damage, including gill lamella proliferation, hepatocyte blurring, and renal necrosis. Transcriptomic analysis revealed that ammonia-N stress promotes the expression of genes related to endoplasmic reticulum stress, heat-shock proteins, immune response, and apoptosis, while inhibiting antioxidant-related genes and Wnt-related genes. Enzymatic assays indicate that ammonia-N stress inhibits the activities of multiple antioxidant enzymes, including SOD, CAT, GSH, GSH-Px, and T-AOC. Microbiome analysis showed that ammonia-N stress altered the intestinal microbial community by increasing harmful bacteria (e.g., Vibrio and Aeromonas) and suppressing beneficial bacteria (e.g., Cetobacterium and Lactococcus). These findings highlight the comprehensive negative impacts of ammonia-N on the health of the spotted longbarbel catfish and provide a theoretical basis for optimizing aquaculture conditions to support the sustainable protection and domestication of the spotted longbarbel catfish.