Plugin error from diversity QIIME 2020.8 PERMANOVA

Hi, I am trying to conduct PERMANOVA on QIMME 2 2020.8 using the following command;
$ qiime diversity beta-group-significance --i-distance-matrix core-metrics-results_11995/unweighted_unifrac_distance_matrix.qza --m-metadata-file GENII_metadata.tsv --m-metadata-column Treatment_day --o-visualization core-metrics-results_11995/unweighted-unifrac-body-site-significance.qzv --p-pairwise --verbose

However, this results in an error message, please see below.
Traceback (most recent call last):
File “/usr/local/bin/miniconda2/envs/qiime2-2020.8/lib/python3.6/site-packages/q2cli/commands.py”, line 329, in call
results = action(**arguments)
File “”, line 2, in beta_group_significance
File “/usr/local/bin/miniconda2/envs/qiime2-2020.8/lib/python3.6/site-packages/qiime2/sdk/action.py”, line 245, in bound_callable
output_types, provenance)
File “/usr/local/bin/miniconda2/envs/qiime2-2020.8/lib/python3.6/site-packages/qiime2/sdk/action.py”, line 452, in callable_executor
ret_val = self._callable(output_dir=temp_dir, **view_args)
File “/usr/local/bin/miniconda2/envs/qiime2-2020.8/lib/python3.6/site-packages/q2_diversity/_beta/_visualizer.py”, line 216, in beta_group_significance
permutations=permutations)
File “/usr/local/bin/miniconda2/envs/qiime2-2020.8/lib/python3.6/site-packages/q2_diversity/_beta/_visualizer.py”, line 122, in _get_pairwise_group_significance_stats
permutations=permutations)
File “/usr/local/bin/miniconda2/envs/qiime2-2020.8/lib/python3.6/site-packages/skbio/stats/distance/_permanova.py”, line 93, in permanova
distance_matrix, grouping, column)
File “/usr/local/bin/miniconda2/envs/qiime2-2020.8/lib/python3.6/site-packages/skbio/stats/distance/_base.py”, line 1025, in _preprocess_input
"All values in the grouping vector are unique. This method cannot "
ValueError: All values in the grouping vector are unique. This method cannot operate on a grouping vector with only unique values (e.g., there are no ‘within’ distances because each group of objects contains only a single object).

Plugin error from diversity:

All values in the grouping vector are unique. This method cannot operate on a grouping vector with only unique values (e.g., there are no ‘within’ distances because each group of objects contains only a single object).

See above for debug info.

Any help in getting this sorted will be helpful. I have not changed the files in any way and using the direct outputs from previous steps as inputs here.

Please let me know if I can provide any further information.

Cheers
Hasinika

Hi @Hasinika,

The error is telling you,

So, at least one of the groups you’re passing only contains one sample. You either need to analyze using a different column or re-group your metadata in some way that you have multiple samples per group.

Best,
Justine