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Kamilari et al. Microbiome Res Rep 2025;4:3 https://dx.doi.org/10.20517/mrr.2024.47 Page 11 of 15

the need for improvement in the hygiene and quality of the products.

Coagulase-negative staphylococci, such as S. succinus, are considered members of the human and animal
[61]
skin microbiota, and their presence is associated with dry fermented sausages . Some strains can perform
[62]
proteolysis and lipolysis, contributing to improved sensory characteristics in sausages . Therefore,
S. succinus presence in traditional Pitsilia sausages may contribute to the flavor and taste of the final
product. Strains that lack the production of staphylococcal enterotoxins and transferable antibiotic
resistance genes are commercially available as starters in fermented sausages . However, their presence in
[63]
human clinical specimens prevents this species from obtaining a qualified presumption of safety (QPS)
status by the European Food Safety Authority . Brochothrix was among the bacterial species that were
[64]
detected in low relative abundances in most Cypriot sausages. Although Brochothrix spp. are non-
pathogenic to humans, they are considered of great importance for the meat products industry due to their
role in causing premature spoilage in refrigerated products . Notably, objectionable odors are produced
[65]
when oxygen concentrations exceed 0.2%. These odors are mostly detected in dry fermented sausages and
are attributed to contamination during packaging operations, as Brochothrix spp. do not survive thermal
processing but can be reintroduced during packaging.

The predominant fungal species of East Mediterranean sausages was D. hansenii. The existence of
D. hansenii is common in fermented sausages and has been found to influence the dynamics of microbial
communities during the ripening of dry sausages in China . Additionally, its proteolytic and lipolytic
[66]
activities positively affect the sensory characteristics of sausages [67-69] . This species possesses a particular
biotechnological importance for the meat products industry due to its nutritional and probiotic effects, in
combination with its increased biotechnological ability [67,70,71] . However, recent studies that correlate
Debaryomyces with inflammation of the gut mucosa and Crohn’s disease create doubts about its addition as
a starter in sausages .
[72]
A common characteristic of Cypriot sausages is the addition of wine. Specifically, in Pitsilia sausages, the
fermentation is performed in local red wine for up to 8 days, followed by smoking. This process might favor
the growth of Saccharomyces. Saccharomyces species are common drivers of alcoholic fermentation during
vinification . Their presence in fermented foods is considered beneficial, not only for improving the
[20]
sensory characteristics of the product but also because of their antagonistic activity against undesirable
microbes. Their metabolic capability to assimilate sugars in grapes to produce ethanol forms a dynamic
stress environment for other members of the microbiota. Furthermore, their ability to secrete antimicrobial
and antifungal compounds, such as myosin, and other killer toxins, further affects the dynamics of the
product’s microbiota [73,74] . Alarcón et al. indicated that the addition of inactive dry Saccharomyces cerevisiae
in “salchichón” sausages as an alternative to sodium ascorbate contributed to increased antioxidant activity,
a steadier phenolic content, and a reduction in protein and lipid oxidation .
[75]

The present study is restricted to only twenty-two samples from Cyprus and eight from Mytilini, which is a
limiting factor in evaluating the selection of microbial biomarkers associated with origin and manufacturing
conditions. Additionally, the geographical distribution of samples should be increased. Sampling should be
conducted across various regions and islands around the Mediterranean to identify the microbial signatures
associated with each region. Moreover, the season of sampling might influence the composition of
microbial communities; therefore, sampling during different seasons will indicate whether microbial
diversity remains consistent. While short-read amplicon 16S rRNA gene sequencing is the primary
approach used for microbiome studies, it cannot differentiate bacteria at the species level. As indicated from
our analysis, bacteria profiling was mostly identified at the genus level, whereas the ITS loci amplicon