*2.4. Statistical Analysis*

Distribution of ARG type in each sample was visualized via Circos graphs, which were carried out by Circos software (http://circos.ca/; accessed on 17 October 2021) online, while Heatmap graphs were produced using RStudio with pheatmap package. The 2−ΔCT abundance table was used to calculate the Pearson Correlation Coefficient and the Bray– Curtis distance. Adonis and Anosim analyses were then conducted using the Bray–Curtis distance matrix. Ordinary least square (OLS) graphs were drawn based on the 2−ΔCT abundance table. The correlation analysis among ARGs and MEGs was performed using the Hmisc package in R. One-way ANOVA test in SPSS version 26.0 was used to analyze the statistical differences of quantity of ARGs between fish farms and the adjacent natural sea area.
