The effect of a high-protein and high-carbohydrate diet on the content of D-lactate in the blood plasma and intestines of a model organism – rainbow trout

Abstract

D-lactic acid stereoisomer (D-lactate) is produced by the intestinal microflora and can enter the bloodstream and cause in some cases a condition of acute D-lactic acidemia known as short gut syndrome. The level of D-lactate in blood and in the contents of the intestine is considered as a promising marker of the development of inflammation associated with microflora disorders, as well as with the development of a bacterial infection, while the mechanism of its entry into the blood of vertebrates from the intestine has not been studied in detail.The aim of the study. To investigate the relationship between the level of D-lactate in blood and in the intestine, taking into account the permeability of the intestinal epithelium.Materials and methods. As a model object of the study, we used juvenile rainbow trout O. mykiss. For 54 days, they were high-carbohydrate or high-protein fed. Since different types of bacteria prefer different substrates, it was expected that at the end of the experiment, the composition of the intestinal microflora would be significantly different in fish fed with different diets. The content of D-lactate in blood plasma in vitro was assessed by the Larsen method with modifications; intestinal permeability was assessed by the intensity of fluorescence of the FITС-Dextran stain in the blood of fish. The analysis of the metagenome of samples of the contents and epithelium of the fore and hind intestine was carried out. The hematological profile was partially characterized using blood smears taken immediately after fish blood sampling. By the means of a different diet, it was possible to obtain two groups of fish that differ significantly in the permeability of the intestinal epithelium and in the content of D-lactate in the intestine. At the same time, despite the differences between the experimental groups in the content of D-lactate in the intestine and in intestinal permeability, no significant differences in D-lactate level in blood were found between them. Analysis of the composition of the intestinal microbiome by metabarcoding for the 16S rRNA gene revealed the absence of lactobacilli in the production of D-lactate in fish.Results. It was shown that the mechanism of accumulation of D-lactate in the blood plasma in fish is less associated with increased intestinal permeability or hyperproduction of this metabolite by the intestinal microflora and is more associated with the utilization of D-lactate in the body. In the experiment, it was not possible to achieve a significant change in the species composition of the intestinal microflora of trout under the influence of a highcarbohydrate diet for 54 days compared to fish that received high-protein diet. Some tendencies towards changes in the composition of the microflora were found in the contents of the hindgut, and perhaps with a longer exposure, these changes could reach a statistically significant level.