lefse analysis error

[KOMALSKK]

Hello, Can you help me with the following error. I created a phyloseq object physeq and performed lefse.

library(microbiomeMarker)
norm_tss(physeq)
normalize(physeq, method = "TSS")

data(physeq)
mm_test <- normalize(physeq, method = "CPM") %>%
run_lefse(
wilcoxon_cutoff = 0.01,
norm = "none", # must be "none" since the input has been normalized
group = "type",
kw_cutoff = 0.01,
multigrp_strat = TRUE,
lda_cutoff = 2

But I get the following output;
mm_lefse <- run_lefse(

• physeq,
• wilcoxon_cutoff = 0.05,
• group = "type",
• kw_cutoff = 0.05,
• multigrp_strat = TRUE,
• lda_cutoff = 2
• )
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
  Also defined by ‘tidytree’
  Error: Too small samples in each class or the variance of feature abundances within a class too small (zero or near zero).

Can you help me finding the error, here.

Thank you.