Analysis of intestinal microbiota of the kuruma shrimp Penaeus japonicus under different bait feedings

This research devised an individual rearing method to compare the effects of live feed (sandworms Perinereis aibuhitensis), formulated pellet diet, and a mixture of live feed and formula feed on the kuruma shrimp Penaeus japonicus, and mitigate the impact of non-food factors on the growth of P. japonicus due to their cannibalism habit. After the feeding experiment, we collected the intestinal tissues of the shrimp for intestinal microbiota analysis. The intestinal tissues of every three 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.