Biowulf High Performance Computing at the NIH
lefse on Biowulf

From the lefse documentation:

LEfSe (Linear discriminant analysis Effect Size) determines the features (organisms, clades, operational taxonomic units, genes, or functions) most likely to explain differences between classes by coupling standard tests for statistical significance with additional tests encoding biological consistency and effect relevance.


  • N. Segata, J. Izard, L. Waldron, D. Gevers, L. Miropolsky, W. S. Garrett, and C. Huttenhower. Metagenomic biomarker discovery and explanation. Genome Biol. 2011, 12:R60. Pubmed |  PMC |  Journal
Important Notes

Interactive job
Interactive jobs should be used for debugging, graphics, or applications that cannot be run as batch jobs.

Allocate an interactive session and run the program. The input format for this tool contains two rows of metadata, one row of sample ids, and a microbial abundance table.

[user@biowulf]$ sinteractive --gres=lscratch:10 --cpus-per-task=2
salloc.exe: Pending job allocation 46116226
salloc.exe: job 46116226 queued and waiting for resources
salloc.exe: job 46116226 has been allocated resources
salloc.exe: Granted job allocation 46116226
salloc.exe: Waiting for resource configuration
salloc.exe: Nodes cn3144 are ready for job

[user@cn3144]$ cd /lscratch/$SLURM_JOB_ID
[user@cn3144]$ module load lefse
[user@cn3144]$ cp ${LEFSE_TEST_DATA:-none}/hmp_aerobiosis_small.txt .
[user@cn3144]$ head hmp_aerobiosis_small.txt  | cut -f1-4
oxygen_availability     High_O2 Mid_O2  Low_O2
body_site       ear     oral    gut
subject_id      158721788       158721788       159146620
Archaea|Euryarchaeota|Methanobacteria|Methanobacteriales|Methanobacteriaceae|Methanobrevibacter 2.96541e-06     5.08937e-06      4.93921e-06
Bacteria        0.999994        0.99999 0.99999
Bacteria|Acidobacteria  5.0412e-05      8.65194e-05     8.39666e-05
Bacteria|Acidobacteria|Acidobacteria_Gp10|Gp10  2.96541e-06     5.08937e-06     4.93921e-06
Bacteria|Acidobacteria|Acidobacteria_Gp11|Gp11  2.96541e-06     5.08937e-06     4.93921e-06
Bacteria|Acidobacteria|Acidobacteria_Gp16|Gp16  2.96541e-06     5.08937e-06     4.93921e-06
Bacteria|Acidobacteria|Acidobacteria_Gp17|Gp17  2.96541e-06     5.08937e-06     4.93921e-06

[user@cn3144]$ hmp_aerobiosis_small.txt\
                  -c 1 -s 2 -u 3 -o 1000000
[user@cn3144]$ hmp_aerobiosis_small.res
f significantly discriminative features: 51 ( 131 ) before internal wilcoxon
Number of discriminative features with abs LDA score > 2.0 : 51

Then plot the LDA scores with

[user@cn3144]$ hmp_aerobiosis_small.res hmp_aerobiosis_small.png --format png --dpi=300
lefse lda scores

Or as a cladogram:

[user@cn3144]$ hmp_aerobiosis_small.res hmp_aerobiosis_small.cladogram.png --format png --dpi 300
lefse cladogram

Copy results back from lscratch and exit

[user@cn3144]$ mkdir -p /data/$USER/lefse_results
[user@cn3144]$ mv ./* /data/$USER/lefse_results
[user@cn3144]$ exit
salloc.exe: Relinquishing job allocation 46116226

Batch job
Most jobs should be run as batch jobs.

Create a batch input file (e.g., which uses the input file ''. For example:

module load lefse/1.0.8 ${LEFSE_TEST_DATA:-none}/hmp_aerobiosis_small.txt \
                  -c 1 -s 2 -u 3 -o 1000000 hmp_aerobiosis_small.res hmp_aerobiosis_small.png --format png --dpi=300 hmp_aerobiosis_small.res hmp_aerobiosis_small.cladogram.png --format png --dpi 300

Submit this job using the Slurm sbatch command.

sbatch --cpus-per-task=2 --mem=5g