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Kamilari et al. Microbiome Res Rep 2025;4:3 https://dx.doi.org/10.20517/mrr.2024.47 Page 9 of 15

Figure 4. The correlation pattern of microbial communities focusing on (A) Lactobacillus; (B) Leuconostoc; (C) Lactococcus; and (D)
Pediococcus. Green and purple edges represent positive or negative correlations between two nodes, respectively, based on Spearman’s
rank correlation, the Pearson correlation, and the Bray Curtis and Kullback–Leibler dissimilarity matrices. The degree of relative
abundance ranges from light to dark blue based on the lower to higher abundant taxa.

hansenii, and Pichia membranifaciens [Figure 4B and C, respectively]. Additionally, Leuconostoc indicated
negative associations, whereas Lactococcus showed positive associations with Streptococcus and
Staphylococcus. Pediococcus, a genus that was detected in some Cypriot sausages, indicated positive
associations with Lactobacillus spp., Leuconostoc spp., and Staphylococcus, and was negatively associated
w i t h t a x a , s u c h a s Enterobacteriaceae, Erysipelotrichaceae, Clostridium, n c l u d i n g C . perfringens,
i
Pseudomonas spp., Pseudobutyrivibrio, Pichia spp., Stemphylium, Kluyveromyces marxianus, and
Kazachstania barnettii [Figure 4D].

DISCUSSION
Emerging evidence highlights the presence of identifiable microbial signatures in fermented products of
different geographic origins and manufactured using different processing conditions [5,20,21,38,39] . This work
analyzed the microbial diversity of fermented eastern Mediterranean sausages from the islands of Cyprus
and Mytilini using metataxonomic sequencing. Apart from characterizing their microbial diversity, the goal
of the study was to identify key microbes that would distinguish between traditionally and industrially
produced Cypriot sausages. Furthermore, focusing on dominant taxa, the study aimed to reveal interactions
that may affect the dynamics of the microbial community composition in sausages, thereby reflecting
sausages’ qualitative characteristics. The study identified distinct microbial signatures in sausages produced
in different geographic areas with different manufacturing conditions (Cypriot traditionally and industrially
produced) using the Lefse biomarkers discovery and random forest algorithms. Specifically, the random
forest algorithm accurately determined the origin and different producing conditions of sausages according
to their microbiota. Both algorithms identified the taxa Lactobacillus and S. succinus as microbial signatures
of traditional Pitsilia sausages and Leuconostoc of Nicosia sausages. In agreement, Kamilari et al. identified