## 4 Discussion  
The ability to monitor ecosystems using non-invasive and relatively inexpensive methods such as eDNA will contribute to the better management of these important habitats and the resources they contain [2; 4; 5; 24; 29]. However, many of the free and open-source tools used for eDNA analysis have complex command-line interfaces that can be challenging for wet lab staff to utilize [30; 31]. The _eDNA-container app_ uses a point-and-click interface that will allow lab scientists to analyze their own eDNA sequencing data using the latest bioinformatics software. The distribution of the app as a _Docker_ image allows for cross-platform usage and supports reproducible eDNA analyses across different computer frameworks. The _eDNA-container app_ automatically generates a PDF report that describes important runtime parameters so that researchers can adjust quality trimming stringency and ASV generation settings. The final ASV spreadsheet includes the DNA sequence of the loci so taxonomic classifications can be confirmed quickly using tools such as _NCBI-BLAST_. Also, alpha diversity and species barplots are reported so scientists can assess the taxonomic sampling depth obtained in the experiment, as well as visualize the species diversity across samples. The _eDNA-container app_ is customizable and can be used to analyze data from any eDNA loci or _QIIME2_-compatible taxonomic database. Importantly, the use of _Docker_ supports reproducible research in the eDNA community, which is an important development as the underlying methodologies continue to be optimized [3; 32]. The development of a free and easy-to-use analysis application will support the increased uptake of eDNA technologies and thus help improve biosecurity and ecosystem monitoring. The following supporting information can be downloaded at: [[https://www.mdpi.com/article/10.3390/app14062641/s1](https://www.mdpi.com/article/10.3390/app14062641/s1)]([https://www.mdpi.com/article/10.3390/app14062641/s1](https://www.mdpi.com/article/10.3390/app14062641/s1)). Supplemental_figs.docx: Supplemental_figs.docx: Supplemental_figs.pdf S1-S3. Supplemental_figs.pdf S1: an example PDF report generated using _the eDNA-container app_. Supplemental_figs.pdf S2: a guide for ecologists and lab scientists on using _Docker_ and _the eDNA-container app_. Supplemental_figs.pdf S3: an example of the eDNA taxonomy counts spreadsheet generated by the app based on the data from [27]. Supplemental_figs.pdf S4: _MiFish_ outputs from the [27] dataset. [PERSON] conceived the idea for the _eDNA-container app_, wrote the software, and drafted the manuscript. [PERSON], [PERSON] and [PERSON] provided critical feedback during testing, contributed \begin{table}
\begin{tabular}{c c c} \hline \hline
**Sample** & **The _eDNA-Container App_** & **MiFish** \\ \hline SRR11479674 & 9 (_Leuciscus idus_) & 10 (_Rhodats anarus_) \\ SRR11479675 & 12 (_Carassius carcass_) & 11 \\ SRR11479676 & 10 & 10 \\ SRR11479677 & 7 & 7 \\ SRR11479678 & 9 & 10 (_Ctenopharynxgodon idella_) \\ SRR11479680 & 9 (_Homo sapiens_) & 8 \\ SRR11479681 & 10 & 10 \\ SRR11479682 & 8 & 8 \\ SRR11479683 & 7 & 7 \\ SRR11479769 & 5 & 6 (_Lissortium vulgaris_) \\ SRR11479770 & 11 & 12 (_Titurus cristatus_) \\ SRR11479771 & 8 (_Fulica atra_) & 7 \\ SRR11479772 & 10 & 10 \\ SRR11479773 & 9 (_Acroccephalus scripaccus_) & 9 (_Ctenopharynxgodon idella_) \\ SRR11479774 & 8 & 8 \\ \hline \hline \end{tabular}
\end{table} 
Table 3: The number of species detected by the _eDNA-container app_ and _MiFish_ based on the data from [PERSON] et al. (2021). When species are only detected by one pipeline, it is indicated in parentheses. Species are only considered if the assignment confidence level is medium/high for _MiFish_ or >90% for the _eDNA-container app_. SRA accession numbers for each sample from [PERSON] et al. (2021) are provided in column 1.  
to the project development, and reviewed the manuscript. All authors have read and agreed to the published version of the manuscript.