Effect of shrimp hepatic enterocytes on intestinal and hepatopancreas flora of the Litopenaeus vannamei

The same batch of Litopenaeus vannamei was put into the aquaculture pond of Ninghai Shengning Aquatic Products Co., Ltd. 30 prawns and 30 shrimps were randomly collected from the same batch of Litopenaeus vannamei, and the intestine and hepatopancreas of prawn were collected under Aseptic technique. Three parallel and sterile cryopreservation tubes were respectively set up and placed in the -80 ℃ refrigerator for standby. Extract the total genomic DNA of the sample using the SDS method. DNA concentration and purity were detected on 1% agarose gel. Dilute the sample DNA to l ug with sterile water according to the concentration/ μ L. Amplification of the variable regions V3 to V4 of the 16 S rRNA gene was performed using forward primers (5 '- GTGCGAGCCGGTAA-3') and reverse primers (5 '- CGCCAG CGCGGGTAA-3'). PCR amplification was performed using a 10 ng DNA sample as a template. The reaction system was pre denatured at 98 ℃ for 1 minute, denatured at 98 ℃ for 10 seconds, annealed at 50 ℃ for 30 seconds, and extended at 72 ℃ for 30 seconds. 30 cycles were conducted, and finally extended at 72 ℃ for 5 minutes and stored at 4 ℃. Use the qiquick PCR purification kit (Qiagen, GmbH, Hilden, Germany) to merge and purify the PCR products of each sample, and use 1% agarose for gel electrophoresis verification of the PCR products. Five sets of samples were sent for testing, including healthy shrimp gut (S.D.C), healthy shrimp hepatopancreas (S.D.G), diseased shrimp gut (S.X.C), diseased shrimp hepatopancreas (S.X.G), and water microbiota (S.S). Finally, sequencing was performed on the Illumina NovaSeq platform.