# LETHAL ERROR: Requested feature \ was missing or not stratified **URL:** \ **Category:** HUMAnN\ **Created:** [June 23, 2023, 1:22pm UTC](https://forum.biobakery.org/t/lethal-error-requested-feature-peptidoglycansyn-pwy-was-missing-or-not-stratified/5524 "2023-06-23T13:22:21Z")\ **Posts on this page:** 4\ **Page:** 1
**Author:** ![Chang\_Jang](https://yyz1.discourse-cdn.com/flex027/user_avatar/forum.biobakery.org/chang_jang/32/2165_2.png) [@Chang\_Jang](https://forum.biobakery.org/u/Chang_Jang)\ **Post date:** [June 23, 2023, 1:22pm UTC](https://forum.biobakery.org/t/lethal-error-requested-feature-peptidoglycansyn-pwy-was-missing-or-not-stratified/5524/1 "2023-06-23T13:22:21Z")
Hello, I have an issue with humann\_barplot with --last-metadata option. When I use the option --last-metadata in humann\_barplot, I have an error message as follows, LETHAL ERROR: Requested feature was missing or not stratified In the abundance table, I checked there is PEPTIDOGLYCANSYN-PWY. I am not sure what “not stratified” means in the error message. Without the option --last-metadata, it is working without an error. Would someone let me know what the problem is? Thanks. ---
**Author:** ![franzosa](https://yyz1.discourse-cdn.com/flex027/user_avatar/forum.biobakery.org/franzosa/32/3511_2.png) [@franzosa](https://forum.biobakery.org/u/franzosa)\ **Post date:** [June 23, 2023, 5:40pm UTC](https://forum.biobakery.org/t/lethal-error-requested-feature-peptidoglycansyn-pwy-was-missing-or-not-stratified/5524/2 "2023-06-23T17:40:07Z")
This script assumes you have potential metadata rows at the top of your table (e.g. sample location, sample time, etc.) followed by the HUMAnN rows. The `--last-metadata` flag tells the script where the metadata ends based on the name of the last metadata feature. I am not sure why using/not using this flag affects your error, but the error you’re seeing means that there are no species stratifications for the pathway you selected, i.e. it only have a community total abundance. This script requires a feature that looks like… ```auto Pathway Pathway|Species1 Pathway|Species2 Pathway|unclassified ``` (for example.) If you don’t have stratified rows for your feature of interest, i.e. rows with headers containing `|`, then the script can’t plot that feature. ---
**Author:** ![Chang\_Jang](https://yyz1.discourse-cdn.com/flex027/user_avatar/forum.biobakery.org/chang_jang/32/2165_2.png) [@Chang\_Jang](https://forum.biobakery.org/u/Chang_Jang)\ **Post date:** [June 26, 2023, 11:18am UTC](https://forum.biobakery.org/t/lethal-error-requested-feature-peptidoglycansyn-pwy-was-missing-or-not-stratified/5524/3 "2023-06-26T11:18:15Z")
Hi Eric, Thanks for your response. I think that I have stratification in _the joined table_. The following is an example of PEPTIDOGLYCANSYN-PWY. PEPTIDOGLYCANSYN-PWY: peptidoglycan biosynthesis I (meso-diaminopimelate containing) PEPTIDOGLYCANSYN-PWY: peptidoglycan biosynthesis I (meso-diaminopimelate containing)|g\_\_Absiella.s\_\_Absiella\_dolichum PEPTIDOGLYCANSYN-PWY: peptidoglycan biosynthesis I (meso-diaminopimelate containing)|g\_\_Acidaminococcus.s\_\_Acidaminococcus\_sp\_CAG\_542 … PEPTIDOGLYCANSYN-PWY: peptidoglycan biosynthesis I unclassified I am not sure why using --last-metadata flag made an error. I will try to figure out what the problem is with comparing a file of hmp\_pathabund.pcl from humann tutorial. Thanks, Chang ---
**Author:** ![franzosa](https://yyz1.discourse-cdn.com/flex027/user_avatar/forum.biobakery.org/franzosa/32/3511_2.png) [@franzosa](https://forum.biobakery.org/u/franzosa)\ **Post date:** [July 7, 2023, 6:33pm UTC](https://forum.biobakery.org/t/lethal-error-requested-feature-peptidoglycansyn-pwy-was-missing-or-not-stratified/5524/4 "2023-07-07T18:33:34Z")
You only need to invoke `--last-metadata` if you have metadata rows at the top of your table that you want to ignore. Is that the case here?