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Page 8 of 15 Kamilari et al. Microbiome Res Rep 2025;4:3 https://dx.doi.org/10.20517/mrr.2024.47

Figure 3. Microbial taxa that can accurately differentiate (A) the bacterial diversity of sausages from different areas; (B) the bacterial
diversity of Cypriot traditionally produced and industrially produced sausages; (C) the fungal diversity of Cypriot traditionally produced
and industrially produced sausages. Heatmap shows the top microbial predictive features that differentiate sausages per each category.
ROC and PCROC analyses for microbial taxa, indicating true and false positive rates for sausages from different areas, industrially or
traditionally produced Cypriot sausages, show perfect predictive accuracy (AUC = 0.99, AUC 0.97, respectively). ROC: Receiver
operating characteristics ; PCROC: per class receiver operating characteristics AUC: area under the curve.

Cladosporium, and Aspergillus ruber, whereas traditional sausages by the species Alternaria tropica,
Cladosporium tenuissimum, Trichosporon otae, Stemphylium vesicarium, and Meyerozyma guilliermondii.

Microbial interaction investigation
To increase our understanding regarding the complicated correlations among the detected taxa, a co-
occurrence network was generated using the CoNet algorithm in Cytoscape [Figure 4]. The interactions
were chiefly oriented toward the keystone taxa that were overrepresented in Cypriot and/or Mytilini
sausages.

Focusing on Lactobacillus, one of the most dominant genera in Cypriot and Mytilini sausages and a
biomarker of traditional Pitsilia sausages, a positive association was observed with Streptococcus,
Lactococcus, and Staphylococcus [Figure 4A]. Negative associations were observed with the taxa
Enterobacteriaceae, Pseudobuturivibrio, Rikenellaceae, Ruminococcaceae, Aspergillus, Candida zeylanoides,
Debaryomyces hansenii, Pichia mandshurica, and Xeromyces bisporus. Both Leuconostoc, a genus that
characterized industrial sausages, and Lactococcus, which characterized sausages from Mytilini, indicated
co-presence with the taxa Pseudomonas, Acinetobacter, Candida zeylanoides, and Brochothrix, whereas
mutual exclusion was observed for Clostridium, Erysipelotrichaceae, Ruminococcacea, Debaryomyces