Effects of different protein and lipid levels on the intestinal microflora of the kuruma shrimp (Penaeus japonicus)

This study aimed at examining the effects of dietary protein and lipid levels on the growth performance and composition of the intestinal microbiota in Penaeus japonicus. A 7-week 3 x 2 two factorial experiment was conducted to investigate the interaction between dietary crude protein (50%, 48%, 46%)and lipid (12%, 6%) on the intestinal microflora of P. japonicus.After the feeding experiment, we collected the intestinal tissues of the shrimp for intestinal microbiota analysis. The intestinal tissues of every 5 shrimps were one sample, and each experimental group had a total of 3 samples. Total bacterial DNA from all shrimp intestines samples was extracted by the CTAB method. The V4-V5 region of 16S rRNA genes was amplified by PCR. Sequencing libraries were generated using TruSeq DNA PCR-Free Sample Preparation Kit (Illumina, USA). The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on Illumina platforms, according to effective library concentration and data amount required.