# Soil Hg Contamination Impact on Earthworms'
## 3 Results
### Beta Diversity in Earthworm, Cast and Soil (Experiment 1)  
In experiment 1, both bacterial and fungal community structures differed significantly between earthworm gut, soil and cast samples (Figure 3, Supplementary Table S3). The Hg-contaminated soil layer greatly affected the earthworm and cast bacterial and fungal community structures but not those of the soil (Table 3). \begin{table}
\begin{tabular}{l c c c c c c c c} \hline \hline
**Variable** & \multicolumn{4}{c}{**Richness**} & \multicolumn{4}{c}{**Shannon Index**} \\ \hline \multicolumn{2}{c}{_sample subset_} & \multicolumn{2}{c}{**Bacteria**} & \multicolumn{2}{c}{**Fungi**} & \multicolumn{2}{c}{**Bacteria**} & \multicolumn{2}{c}{**Fungi**} \\ \hline  & **DF** & **F** & **P** & **F** & **P** & **F** & **P** & **F** & **P** \\ \hline
**Experiment 1** & & & & & & & & & \\ Type of sample & 2, 29 & 7.5 & 0.003 & 22.24 & \textless{}0.001 & 3.8 & 0.036 & 5.2 & 0.124 \\ Treatment & 1, 29 & 13.7 & 0.001 & 31.49 & \textless{}0.001 & 15.4 & \textless{}0.001 & 3.8 & 0.062 \\ \multicolumn{2}{c}{_earthworm samples_} & 1, 11 & 107 & \textless{}0.001 & 246.9 & \textless{}0.001 & 868.5 & \textless{}0.001 & 41.5 & \textless{}0.001 \\ \multicolumn{2}{c}{_cast samples_} & 1, 11 & 4.4 & 0.062 & 12.23 & 0.006 & 1.0 & 0.339 & 0.0 & 0.862 \\ \multicolumn{2}{c}{_soil samples_} & 1, 5 & 9.7 & 0.036 & 3.51 & 0.134 & 1.4 & 0.303 & 1.3 & 0.313 \\ \hline
**Experiment 2** & & & & & & & & & \\ Hg treatment & 1, 49/87 * & 22.1 & \textless{}0.001 & 0.4 & 0.531 & 34.0 & \textless{}0.001 & 2.4 & 0.127 \\ \multicolumn{2}{c}{_soil samples_} & 1, 3/7 * & 18.6 & 0.050 & 0.1 & 0.718 & 259.9 & 0.004 & 0.1 & 0.743 \\ \multicolumn{2}{c}{_stomach samples_} & 1, NA/15 * & NA & NA & 0.5 & 0.500 & NA & NA & 3.0 & 0.103 \\ \multicolumn{2}{c}{_UgT samples_} & 1, 5/15 * & 0.6 & 0.500 & 0.1 & 0.729 & 0.9 & 0.401 & 0.5 & 0.480 \\ \multicolumn{2}{c}{_UgS samples_} & 1, 7/15 * & 9.5 & 0.022 & 0.3 & 0.609 & 8.4 & 0.027 & 0.2 & 0.650 \\ \multicolumn{2}{c}{_LgT samples_} & 1, 14/15 * & 18.0 & 0.001 & 0.0 & 0.828 & 9.6 & 0.008 & 0.0 & 0.896 \\ \multicolumn{2}{c}{_LgS samples_} & 1, 14/15 * & 1.9 & 0.192 & 0.0 & 0.869 & 0.3 & 0.618 & 0.1 & 0.736 \\ Earthworm gut part & 5, 49/87 * & 7.4 & \textless{}0.001 & 6.7 & \textless{}0.001 & 8.4 & \textless{}0.001 & 3.3 & 0.009 \\ \hline \hline \end{tabular} * for bacteria/fungi.  
\end{table} 
Table 2: Statistical differences (ANOVA) in bacterial and fungal richness and Shannon diversity index between Hg treatments and among sample types in experiment 1 and between Hg treatments and among earthworm gut parts in experiment 2. DF = Degrees of freedom. _Appl. Sci._**2020**, _10_, 2565  
Figure 3: Beta diversity of the bacterial (**A**) and fungal (**B**) communities in experiment 1, presented in a non-metric multidimensional (NMDS) ordination including the three sample types (earthworm, cast and soil) and the two treatments (uncontaminated and Hg-contaminated). \begin{table}
\begin{tabular}{c c c c c c} \hline \hline
**Variable** & \multicolumn{3}{c}{**Bacteria**} & \multicolumn{3}{c}{**Fungi**} \\ \hline _Sample subset_ & **DF** & **F** & **P** & **F** & **P** \\ \hline
**Experiment 1** & & & & & \\ Type of sample & 2, 29 & 8.3 & \(<\)0.001 & 4.6 & \(<\)0.001 \\ Hg Treatment & 1, 29 & 5.9 & \(<\)0.001 & 3.6 & 0.001 \\ _earthworm samples_ & 1, 11 & 21.5 & 0.002 & 5.0 & 0.002 \\ _cast samples_ & 1, 11 & 4.4 & 0.002 & 12.3 & 0.002 \\ _soil samples_ & 1, 5 & 1.0 & 0.400 & 0.9 & 0.700 \\ \hline
**Experiment 2** & & & & & \\ Hg treatments & 1, 49/87 * & 6.5 & \(<\)0.001 & 4.0 & \(<\)0.001 \\ _soil samples_ & 1, 3/7 * & 2.5 & 0.333 & 1.4 & 0.028 \\ _stomach samples_ & 1, NA/15 * & NA & NA & 1.2 & 0.114 \\ _UgT samples_ & 1,5/15 * & 1.3 & 0.333 & 2.5 & 0.003 \\ _UgS samples_ & 1,7/15 * & 1.5 & 0.168 & 1.7 & 0.045 \\ _LgT samples_ & 1,14/15 * & 1.2 & 0.307 & 1.8 & 0.007 \\ _LgS samples_ & 1,14/15 * & 1.0 & 0.406 & 1.0 & 0.379 \\ Earthworm gut part & 5, 49/87 * & 2.8 & \(<\)0.001 & 2.1 & \(<\)0.001 \\ \hline \hline \end{tabular}
* for bacteria/fungi.  
\end{table}