Synbiotics, a synergistic combination of probiotics and prebiotics, are currently regarded as one of the most practical nutritional supplements in tilapia farms. In this study, the effect of supplementing the diet of red tilapia (Oreochromis spp.) with Jerusalem artichoke (Helianthus tuberosus) and Lactobacillus rhamnosus GG (LGG) was evaluated. Growth performance, serum biochemical parameters, intestinal morphology, goblet cell counts, immune parameters and protection against Aeromonas veronii challenge were determined. The results showed that fish fed with synbiotic-supplemented diets had a significantly higher (P < 0.05) feed conversion ratio (FCR), specific growth rate (SGR), and average daily gain (ADG) than fish fed with a control diet. The synbiotic-supplemented diet increased glucose, total protein and the total cholesterol levels. The absorptive area of the proximal and distal intestine of fish fed on the synbiotic diet was significantly higher (P < 0.05) than in those fed with probiotics (LGG), prebiotic-supplemented diets (JA), and the control diet. Goblet cell counts revealed that the numbers of acid mucous cells, neutral mucous cells and double-staining mucous cells of fish fed the synbiotic-supplemented diet (JA + LGG) were significantly higher (P < 0.05) in the proximal and distal intestine. Fish fed the synbiotic-supplemented diets also exhibited significantly higher (P < 0.05) lysozyme activity. The cumulative mortalities of fish fed with a synbiotic-supplemented diet were significantly lower than those of fish fed other diets. The results suggested the beneficial effect of JA and LGG synbiotic diet on growth performance and health status of red tilapia. Direct administration of JA and LGG in fish feed can be used as a practical nutritional supplement in red tilapia. Copyright © 2018 Elsevier Ltd. All rights reserved.