| Summary: | During Pecorino Romano PDO cheese production, <i>scotta</i> (residual whey from <i>ricotta</i> cheese manufacturing) or <i>siero</i> (whey) can be integrated with autochthonous starters, natural and composed of an indefinite number of species and strains, or commercial selected starters to obtain <i>scotta/siero-innesto</i>. In this study, three biodiverse autochthonous natural starter cultures (SR30, SR56, and SR63) belonging to the Agris Sardegna BNSS microbial collection, composed of different strains belonging to the species <i>Streptococcus thermophilus</i>, <i>Lactobacillus delbrueckii</i> subsp. <i>bulgaricus</i>, <i>Enterococcus faecium</i>, and <i>Limosilactobacillus reuteri</i> were lyophilized and combined into two starter mixes (A and B). The <i>scotta/siero-innesto</i> and the Pecorino Romano obtained using natural starters were compared with those obtained using commercial selected starters during three seasons of the cheesemaking campaign. Different pH and microbial compositions for the <i>scotta/siero-innesto</i> obtained using natural or commercial starters were found, attributable to their different biodiversity. The six-month-ripened cheese microbiota was influenced mostly by the season of cheesemaking, whereas physico-chemical and sensory analyses did not highlight differences among the products obtained. In general, no effect attributable to the type of <i>scotta/siero-innesto</i> used was observed, allowing the conclusion that natural starter cultures can be used also in industrial-scale production, ensuring high stability in the technological performances and preserving the microbial, chemical, and sensory characteristics of Pecorino Romano PDO cheese.
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