Bighead carp (), an important cultured freshwater fish in China, is one of the four major Chinese carps. The natural habitation of bighead carp in the Yangtse River has been destroyed because of environmental water pollution and overfishing, among other factors. Stock enhancement of bighead carp has been conducted for years, being regarded as a positive fisheries management tool. Currently, hatchery-reared bighead carp juveniles were released into the natural river system to supplement the wild population in the Yangtse River. However, the effectiveness of these enhancement efforts has not be evaluated to date. The application of microsatellite markers for parentage determination is becoming more acceptable and popular in aquaculture. To analyze the effect of stock enhancement of bighead carp, a parentage test based on SSR markers was applied in this study. Fins of 563 brood stocks of unknown sexes and 687 recaptured individuals were sampled. Ten SSR markers were modified using fluorescent dyes (FAM/HEX). Genotype data of all brood stocks and all offspring were screened using an ABI Prism3 730 sequencer. The results showed that the number of alleles of microsatellite markers ranged from 19 to 55, and the mean observed and expected heterozygosities were 0.748 and 0.882. The range of the polymorphism information content (PIC) was 0.797-0.924. The simulation of parentage assignment for the situation of unknown sexes showed the combined non-exclusion probability of 10 loci was 99.996% using Cervus3.0. The results showed that the 42 recaptured individuals were offspring of brood stocks. The study suggested that the total contribution of the brood stocks to the wild population was 6.11% in the Jiangsu reaches of the Yangtse River. In summary, this study established an evaluation system of "marking brood stocks-releasing offspring-recapturing-parentage assignment" to assess the contribution rate for the large-scale stock enhancement. The evaluation system was used in this study to monitor all offspring of the marked brood stocks, which achieved the goal of one-time labeling and multi-year monitoring. A microsatellite gene bank was constructed using 10 microsatellite loci for brood stocks from 2016 to 2018. Future research should pay more attention to the assessment of genetic differentiation between the released population and wild population. This study will provide technical support for the stock enhancement process of bighead carp.