Dietary Effects of Lactobacillus plantarum Combined with Galactooligosaccharide on Immunological and Biochemical Parameters, Gut Microbiota, Digestive Enzyme Activity, Body Composition, and Stress Resistance in Narrow-Clawed Crayfish, Pontastacus leptodactylus (Eschscholtz, 1823)

Abstract

The effects of galactooligosaccharide (GOS) and varying levels of Lactobacillus plantarum (LP) on some physiological parameters as well as air exposure stress resistance in clawed crayfish (Pontastacus leptodactylus) were investigated. During a 97-day trial, 216 crayfish were fed with four experimental diets (in triplicates) including the control diet (without GOS and LP), GLP7 (GOS 2%+LP 107 CFU.g-1 diet), GLP8 (GOS 2%+LP 108 CFU.g-1 diet), and GLP9 (GOS 2%+LP 109 CFU.g-1 diet). At the end of the trial, 26% higher amounts of the total hemocyte count (THC) and semigranular cells (SGC) and 27% higher hyaline cells (HC) were observed in GLP7. A significant improvement in lysozyme value was observed in GLP7 and GLP8. Moreover, superoxide dismutase was significantly higher in GLP9, whereas the catalase (CAT) activity did not change in the experimental groups. Unlikely, plasma glucose levels decreased in all the GLP treatments. In terms of intestinal microbiota, autochthonous lactic acid bacteria (LAB) remarkably increased in all the GLP-supplemented groups, while total autochthonous intestinal heterotrophic bacteria counts (TVC) did not change in GLP-supplemented groups. However, 14 days after switching to the basal diet, no reduction was detected in TVC, but LAB levels decreased in GLP7 and GLP9 treatments. Dietary administration of GLP could beneficially modulate digestive enzyme activity, including protease, lipase, and amylase. Furthermore, higher lipid and gross energy were observed in GLP9. However, GLP-supplemented diets could not improve growth performance parameters. After 24 h subjecting to air exposure stress, although no mortality was detected, crayfish fed GLP-supplemented diets were healthier through enhanced THC, SGC, and HC levels in GLP7 and GLP9 and CAT activity in GLP7 and GLP8. These results proved the positive impacts of dietary supplementation with combined GOS and LP, with GLP8 and GLP9 as optimum diets, on immunochemical parameters, intestinal microbiota, digestive enzyme activity, and stress resistance in P. leptodactylus.