Rosmarinic acid alone or in combination with Lactobacillus rhamnosus ameliorated resistance to ammonia stress in the rainbow trout, Oncorhynchus mykiss: growth, immunity, antioxidant defense and liver functions

Rosmarinic acid (RS) and Lactobacillus rhamnosus (LR) were added singularly or in combination to rainbow trout (Oncorhynchus mykiss) diets to test their efficacy in the protection against ammonia stress. Fish (31.4±0.6 g) were randomly allocated to six groups in three replicates, as follows: T₁: basic food as control, T₂: LR with a concentration of 1.5 × 10⁸ CFU /g, T₃: LR with a concentration of 3 × 10⁸ CFU/g, T₄: 1 g RS/kg, T₅: 3 g RS/kg, and T₆: 1.5 × 10⁸ CFU/g LR + 1 g RS/kg and T₇: 3 × 10⁸ CFU/g LR + 3 g RS/kg. After 60 days feeding, fish were exposed to ammonia stress. After the feeding period, the supplemented fish had the highest final body weight (FW), weight gain (WG), and specific growth rate (SGR), and the lowest feed conversion ratio (FCR) as compared with the control group (P<0.05). Amylase, protease and lipase activities were noticed markedly higher in fish supplemented with 1.5 × 10⁸ CFU/g LR + 1 g RS/kg and 1.5 × 10⁸ CFU/g LR diets compared to the control (P<0.05). Generally, fish in supplemented diets, particularly T₂ and T₆ groups, had the highest lysozyme, alternative complement activity (ACH₅₀), total Ig, nitroblue tetrazolium test (NBT), myeloperoxidase (MPO), complement component 3 (C3), complement component 4 (C4), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx). On the other hand, T₂ and T₆ groups had the lowest malondialdehyde (MDA), glucose, and cortisol concentrations as well as alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) enzyme levels when compared with the control (P<0.05). After ammonia stress, fish in the supplemented groups, particularly T₂ and T₆, generally showed significantly higher values of lysozyme, ACH50, total Ig, NBT, MPO, C3, C4, SOD, CAT, GPx and lower levels of MDA, glucose, cortisol, ALT, ALP, LDH when compared with the control (P<0.05). In conclusion, a combined administration of RS and L. rhamnosus effectively improved growth performance and health status as well as enhanced the resistance of rainbow trout against ammonia toxicity.