Application of ad hoc transformation protocols and selected lactic acid bacteria for the innovation in the ewes’ milk dairy production sector

L'innovazione riveste un ruolo fondamentale in vari settori economici e i prodotti lattiero-caseari esercitano un’influenza significativa sul mercato alimentare. In Italia la secolare tradizione di produrre formaggi di pecora ha guadagnato nuova rilevanza. I formaggi ovini rappresentano non solo un’attività economica primaria, ma fanno anche da volano per l’industria del turismo. Essi contribuiscono a valorizzare i prodotti locali, preservare il patrimonio culturale e apprezzare i paesaggi naturali. Di conseguenza, la produzione di formaggi si è affermata come un motore critico per lo lo sviluppo, la diversificazione e l'innovazione. In Sicilia, una regione euro-mediterranea, si promuove attivamente la trasformazione di questo latte in diverse varietà di formaggio. Tuttavia, il settore lattiero-caseario dei piccoli ruminanti affronta ostacoli organizzativi. Molte aziende casearie si concentrano sulla riproduzione e sulla produzione di formaggi di nicchia, affidandosi a tecniche e attrezzature tradizionali. Nonostante queste sfide, l’innovazione settoriale rimane essenziale per la sopravvivenza economica delle aziende casearie.Tenendo conto di queste considerazioni, all’interno del contesto di un programma di dottorato triennale, si è giunti a una decisione per promuovere l’industria lattiero-casearia ovina. L’obiettivo principale era ampliare la gamma prodotti caseari siciliani introducendo formaggi innovativi ad elevato valore nutrizionale e organolettico per i consumatori. Per raggiungere questo obiettivo, si è proceduto con l’aggiunta di sostanze polifenoliche, da tempo riconosciute per le proprietà antiossidanti benefiche per il corpo umano, nonché di ingredienti quali oli essenziali, che agiscono come conservanti naturali grazie ai loro effetti antimicrobici. Questo approccio strategico non solo ha affrontato la crescente domanda dei consumatori riguardo alla trasparenza sull’origine delle materie prime sulla qualità e sulla sicurezza, ma ha anche aperto la strada a protocolli innovativi. Tali protocolli sono stati appositamente progettati per la produzione di formaggi freschi a base di latte ovino pastorizzato, garantendo il rispetto di rigorosi standard igienici e microbiologici. Inoltre, si sono estesi alla creazione di prodotti lattiero-caseari fermentati, come lo yogurt e i formaggi stagionati, utilizzando sia il latte crudo che quello pastorizzato. Sfruttando starter microbici attentamente selezionati, è stata condotta un’Analisi del Sequenziamento del Genoma Completo (WGSA) per caratterizzarne in modo completo i modelli genetici. Questo approccio ha permesso un rigoroso controllo di qualità e sicurezza. Attraverso diversi articoli di ricerca, i risultati hanno costantemente dimostrato la sicurezza igienico e sanitaria dei prodotti finiti. Inoltre, si è registrato un notevole miglioramento delle caratteristiche nutrizionali, tra cui gli acidi grassi e i composti organici. Gli assaggiatori, valutando questi prodotti a livello sensoriale, hanno espresso un apprezzamento generale. Allargando la prospettiva, questa ricerca getta luce su come l'innovazione dei prodotti, nel settore primario possa influenzare lo sviluppo rurale e le economie locali, specialmente nelle aree marginali. Ad esempio, attraverso il turismo, il formaggio rappresenta un forte elemento della cultura culinaria e dell’identità culturale. I consumatori moderni apprezzano sempre più l’autenticità e il legame genuino con il mondo che il formaggio rappresenta.Innovation plays a pivotal role across various economic sectors, and dairy products hold significant sway in the food market. In Italy, age-old tradition of making cheese from ewe's milk has gained renewed prominence. Ewe’s milk cheeses now serve not only as a primary economic pursuit but also as vital contributors to the tourism industry. They help showcase local products, preserve cultural heritage, and celebrate natural landscapes. Consequently, cheese production has emerged as a critical driver for rural development, diversification, and innovation. Sicily, a Euro-Mediterranean region, actively promotes the transformation of this milk into different cheese varieties. However, the small ruminant dairy sector faces organisational hurdles. Many dairies focus on the breeding and niche-market cheese production, relying on traditional techniques and equipment. Despite these challenges, sectoral innovation remains essential for the economic survival of small dairy farms. Given these considerations, within the context of a three-year PhD program, a decision was reached to advance the ovine dairy industry. The primary objective was to expand the range of Sicilian dairy offerings by introducing novel cheeses with significant nutritional and sensory value for consumers. Achieving this involved incorporating polyphenolic compounds, which have long been recognized for their beneficial antioxidant properties in human body, as well as ingredients like essential oils that act as natural preservatives due to their antimicrobial effects. This strategic approach not only addressed the growing consumer demand for transparency regarding raw material origin, quality, and safety, but also paved the way for innovative protocols. These protocols were specifically designed for producing fresh cheeses from pasteurized ewe's milk, ensuring compliance with rigorous hygienic and microbiological standards. Additionally, they extended to the creation of fermented dairy products such as yogurt and ripened cheeses, using raw and pasteurized milk. Utilizing carefully selected microbial starters, a Whole Genome Sequencing Analysis (WGSA) was conducted to characterize their genetic patterns comprehensively. This approach allowed for rigorous quality and safety control. Across multiple research papers, the results consistently demonstrated the hygienic and sanitary safety of the final dairy products. Additionally, there was notable improvement in nutritional attributes, including fatty acids and organic compounds. Judges, evaluating these products on a sensory level, expressed overall satisfaction. Zooming out, this research sheds light on how product innovation within the primary sector can impact rural development and local economies, especially in marginal areas. For instance, through tourism, cheese serves as an embodiment of gastronomic culture and cultural identity. Modern consumers increasingly appreciate the authenticity and genuine connection to the world that cheese represents

Phytochemical profiling and investigation of antioxidant, anti-proliferative, and antibacterial properties in spontaneously grown Sicilian sumac (Rhus coriaria L.) fruits

This study aimed to assess the phytochemical profile and associated functional properties of sumac (Rhus coriaria L.) fruits harvested from wild plants in Sicily. Chemical characterization unveiled exceptionally high levels of polyphenols (10.99 g GAE/100 g DW), including substantial quantities of proanthocyanidins (61.577 mg PACE/100 g DW), and identified 82 phytochemicals belonging to various flavonoid classes. The hydroalcoholic extract from Sicilian sumac exhibited remarkable redox-active properties, providing antioxidant protection in a cell-based model of lipid peroxidation (CAA50: 1.116 μg/mL). Additionally, it displayed significant antiproliferative activity against four human tumor epithelial cell lines with GI50 values ranging from 31.08 to 149.74 μg/mL and robust antibacterial activity against major foodborne pathogens (MIC: 12.5–25.0 mg/mL). Our findings highlight Sicilian sumac fruit as a rich source of phytochemicals that positively contribute to the cellular redox state even when consumed in small quantities. Additionally, its diverse bioactivities indicate potential applications across food and non-food sectors

Description of Ewiss cheese, a new ewe milk cheese processed by Swiss cheese manufacturing techniques: Microbiological, physicochemical, and sensory aspects

Typically, Swiss-type cheese is made from cow milk. However, in the present work an attempt to expand the sheep supply chain and product offering in this field was made by developing a new type of cheese using Swiss- type cheese technology. The cheese was manufactured under industrial conditions, and fermentations were carried out using freeze-dried commercial starters that are traditionally used in the production of Swiss cheese. Two experimental "Ewiss cheese" (EC) products were produced using raw milk (RM) and pasteurized milk (PM), respectively. Fourteen microbial groups were investigated by plate counts from curd until ripened cheeses. According to microbiological analyses, no statistically significant differences were found between the 2 productions with respect to the group of lactic acid bacteria (LAB). The curds were mainly characterized by mesophilic LAB cocci (7.45 log10 10 cfu/g in RM-EC and 7.33 log10 10 cfu/g in PM-EC). However, at the end of the ripening period (9 mo), the cheeses exhibited a higher presence of mesophilic LAB rods. Undesired microbiological groups were found only in the curd of raw milk cheese in the range of 104 4 to 105 5 cfu/g, but they were reaching undetectable levels by plate count in the cheese at the end of ripening. The RMEC and PM-EC were characterized by 76% and 68% of DM, respectively. These cheeses contained 29.30% and 34.36% of protein, and 51.31% and 50.38% of fat, respectively. Textural analysis showed differences in terms of hardness, chewiness, and gumminess between the experimental cheeses and Swiss cheese sold on the market. These differences could be attributed to the higher protein content of ewe milk. The main fatty acids in the cheeses were palmitic acid, myristic acid, oleic acid, and capric acid. Among the organic acids, RMEC had higher concentrations of lactic acid, whereas PM-EC was higher in propionic acid. The ewe cheeses emitted 46 volatile compounds, including acids, aldehydes, ketones, esters, alcohols, and other compounds. The PM-EC was characterized by the main compounds of Swiss-type cheese: acetic acid, butyric acid, ethyl butyrate, ethyl caproate, propanoic acid, and tetramethylpyrazine. Sensory evaluation showed that the new dairy products were generally appreciated, and PM-EC was the most preferred by the judges. This research has enabled the development of new ewe milk products, which could stimulate the valorization of a sector that has been long neglected and still has a large margin of improvement

The wooden shelf surface and cheese rind mutually exchange microbiota during the traditional ripening process

The rind acts as a protective barrier for internally-bacterial ripened cheeses. Unlike surface-inoculated smear cheeses, centripetal maturation is not assumed to occur in these cheeses. This research was aimed to evaluate the microbial diversity of the wooden shelves used for the ripening of Protected Denomination of Origin (PDO) Pecorino di Filiano and Protected Geographical Indication (PGI) Canestrato di Moliterno cheeses. The microorganisms associated with the rind of these cheeses were also investigated. Both wooden shelf surfaces and cheese rinds were sampled by brushing method to collect their biofilms. Wooden shelves showed levels of total mesophilic microorganisms (TMM) between 5.6 and 7.2 log CFU/cm2, while cheese rinds between 6.1 and 7.8 log CFU/cm2. The major dairy pathogens (Salmonella spp., Listeria monocytogenes, Escherichia coli, and Staphylococcus aureus) were never detected, while mesophilic and thermophilic bacteria dominated the surfaces of all wooden shelves and cheese rinds. LAB community was represented by Enterococcus spp., Leuconostoc spp., and Marinilactibacillus spp. Among yeasts, Debaryomyces spp., Candida spp., were identified, while Aspergillus spp., and Penicillium spp., dominated the community of filamentous fungi. MiSeq Illumina analysis identified 15 phyla, 13 classes, 28 orders, 54 families, and 56 genera among bacteria. Staphylococcus spp. was identified from all wooden surfaces, with a maximum abundance of 71 %. Brevibacterium, Corynebacterium and halophilic bacteria were detected in almost all samples. Regarding fungi, wooden shelves mainly hosted Aspergillus, Penicillium and Debaryomyces hansenii, while cheese rinds especially Penicillium and D. hansenii. Alpha diversity confirmed a strict correlation between the microbiota of wooden shelves and that of cheese rinds for the majority of factories. This study confirmed that the wooden shelves used for cheese ripening are microbiologically active and represent safe systems. Furthermore, the results of this work clarified the transfer flow between wooden shelves and PDO Pecorino di Filiano and PGI Canestrato di Moliterno cheese surfaces: smear-active microorganisms are mainly transferred from wooden shelves to cheese rind, which potentially contribute to the development of the final organoleptic characteristics; meanwhile, cheeses transfer LAB that are potentially involved in defining the safety aspects of the shelve

Addition of fruit purees to enhance quality characteristics of sheep yogurt with selected strains

The aim of this research was to assess the effects of fruit purees of banana, kiwi, mango, red berry, and strawberry on the microbiological, physicochemical, antioxidant, and sensory properties of sheep yogurt. The fruit purees were characterized for their microbiological profile before yogurt production, and no spoilage or pathogenic microorganisms were detected in any of the purees analyzed. Yogurt productions were carried out under industrial conditions using pasteurized sheep’s milk and selected starter cultures of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. Five experimental yogurt productions (EYP) were made by adding 10 % (w/w) of each fruit puree, while the control yogurt production (CYP) was puree-free. Plate counts revealed that levels of viable Lb. delbrueckii subsp. bulgaricus and S. thermophilus exceeded 8.0 log cfu/g in all CYP and EYP samples after 3 d of refrigerated storage. The addition of fruit purees reduced fat percentage until to 7 % and increased of antioxidant activity, especially with red berry puree. Except for banana, the addition of fruit purees resulted in a statistically significant increase (p < 0.0001) in the total terpene VOC profiles of EYP. Notably, the terpene content in mango-flavored yogurt was eightfold greater than that observed in the control trial. Sensory evaluation revealed a reduction in unpleasant odor and off-flavor, and an increase of about 50 % in overall acceptance for all EYP in comparison to CYP. Therefore, adding fruit purees to sheep yogurt is a promising strategy for the valorization of Sicilian sheep’s milk

Description of “Ovino Belmontese”, a new semisoft sheep's milk cheese processed using “Italico” cheese technology

The objective of this study was to create a new semisoft sheep’s milk cheese called “Ovino Belmontese” cheese (OBCh) by applying the “Italico” cheese-making technology. The cheese production took place under industrial conditions, with the addition of a commercial starter formulation containing Streptococcus thermophilus. The microbiological, physicochemical, and sensory characteristics of OBCh were assessed and compared to those of a commercially available cow’s Italico cheese (CICh). Streptococcus thermophilus dominated the microbial community during the cheese-making process, reaching levels of approximately 9.0 Log CFU/g in both OBCh and CICh. Among physical characteristics, no statistically significant difference (p ≥ 0.05) was registered in terms of lightness, redness, yellowness, and hardness between the two cheeses. OBCh exhibited a twofold higher short-chain fatty acid content compared to CICh. Both cheeses displayed similar classes of volatile organic compounds, although their relative percentages differed. The application of Italico cheese technology to process sheep’s milk did not negatively affect sensory attributes. This study highlighted that utilizing a cheese-making technology not commonly used for processing sheep’s milk represents a promising strategy to diversify Sicilian dairy productions

Polyphasic characterization of microbiota of “Mastredda”, a traditional wooden tool used during the production of PDO Provola dei Nebrodi cheese

11openInternationalItalian coauthor/editorThe biofilms of the wooden tables used for the acidification of the curd were investigated for PDO Provola dei Nebrodi cheese, a traditional stretched cheese made in eastern Sicily (southern Italy) from raw cows’ milk. To this purpose the wooden tables of four dairy facilities were analysed for their microbiota by scanning electron microscopy (SEM) analysis and a combined culture-independent and -dependent microbiological approach. SEM inspection showed an almost continuous biofilm formation. MiSeq Illumina analysis identified 8 phyla, 16 classes, 25 orders, 47 families and 50 genera. Corynebacterium, Bifidobacterium and lactic acid bacteria (LAB) were detected in all samples. In particular, the LAB genera detected on all wooden tables were Lactobacillus, Streptococcus and Lactococcus. LAB dominated the surfaces of all wooden tables with levels higher than 7.0 Log CFU/cm2. In particular, the LAB found at the highest levels were mesophilic cocci. Coagulase positive staphylococci, Salmonella spp., Listeria monocytogenes and Shiga-toxigenic Escherichia coli were never detected. Twenty-seven dominating LAB strains were identified within the genera Enterococcus, Lactobacillus, Lacticaseibacillus, Lactiplantibacillus, Levilactobacillus, Lactococcus, Leuconostoc, Pediococcus and Streptococcus. This work showed that the wooden table used during the production of PDO Provola dei Nebrodi cheese is a safe system and a microbiologically active toolopenBusetta, Gabriele; Garofalo, Giuliana; Mangione, Guido; Botta, Luigi; Franciosi, Elena; Di Gerlando, Rosalia; Todaro, Massimo; Licitra, Giuseppe; Scatassa, Maria Luisa; Gaglio, Raimondo; Settanni, LucaBusetta, G.; Garofalo, G.; Mangione, G.; Botta, L.; Franciosi, E.; Di Gerlando, R.; Todaro, M.; Licitra, G.; Scatassa, M.L.; Gaglio, R.; Settanni, L

Reduction of PDO Pecorino Siciliano cheese making duration: microbial dynamics and quality attributes deriving from replacing whey permeate with hot water during cooking

This work was carried out with the aim to reduce the transformation duration of Protected Designation of Origin (PDO) Pecorino Siciliano cheese. To this purpose, the cooking in hot water (experimental production, EXP) was compared to the traditional cheese cooking under whey permeate (control production, CTR). The microbiological composition of under rind (UR) and core (Co) section of CTR and EXP cheeses was determined by a combined culture-dependent and -independent approach. Total mesophilic microorganisms and lactic acid bacteria (LAB) present in raw ewes' milk (5.0 log CFU/mL) increased during cheese making and reached values of about 8.0 log CFU/g in both sections (UR and Co) of 5-month ripened cheeses of both productions (CTR and EXP) monitored. The identification of the viable LAB populations in ripened cheeses showed that Enterococcus, Lacticaseibacillus, Lactiplantibacillus, Levilactobacillus, Limosilactobacillus and Streptococcus dominated UR and Co sections of all cheeses. MiSeq Illumina analysis demonstrated that LAB populations (lactobacilli, lactococci and streptococci) dominated the bacterial community of cheeses at 95.63-98.41 % of relative abundance. The two different cooking operations did not influence the physicochemical characteristics of PDO Pecorino Siciliano cheeses. Sensory evaluation performed by artificial senses analysis and trained panelists confirmed that the modification of PDO Pecorino Siciliano cheese production protocol did not significantly affect product characteristics and overall acceptance. Thus, data of this work confirmed that cooking under hot water allowed to reduce transformation duration and safeguard typicality of PDO Pecorino Siciliano cheese

Comprehensive analysis of Moringa oleifera leaves’ antioxidant properties in ovine cheese

This study aimed to enhance “Pecorino” type ovine cheese by adding Moringa oleifera leaves powder (MOLP). Cheese-making trials, conducted at industrial level, used raw ewes’ milk and two selected Lactococcus lactis strains. The experimental plan included a control production (CTR), and two experimental productions with 1% or 2% MOLP addition (1-MOLP and 2-MOLP, respectively). MOLP did not hinder starters development, which reached about 8.0 Log CFU/g in 2-month ripened cheeses. Illumina results highlighted lactococci dominance in all trials [45.98%–62.48% of relative abundance (RA)]. Physicochemical analysis showed that MOLP-enriched cheeses had higher protein content and lower secondary lipid oxidation. The addition of MOLP increased total phenolic compounds in cheese, reaching 3.64 mg GAE/g in the 2-MOLP sample. MOLP-enriched cheeses showed significantly higher radical scavenging activity than CTR production (p < 0.0001). Ultrahigh-performance liquid chromatography-electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) revealed increased levels of chlorogenic acid, protocatechuic acid, caffeic acid, and ferulic acid due to MOLP enrichment. In the presence of MOLP, cheese volatile organic compounds were affected by compounds like 2-octanone, 3-hexen-2-one, heptane, nonanol, and linalool. 1-MOLP cheese was comparable to CTR production in overall satisfaction (sensory evaluation). Including MOLP in cheese production offers exciting opportunities for functional Sicilian ewes’ milk products

Development of “Quadrello di Ovino”, a Novel Fresh Ewe’s Cheese

This work was performed to produce a new soft ewe’s milk cheese, namely “Quadrello di ovino” (QdO) cheese, to enlarge ewe’s dairy product portfolio of South Italy, barely limited to Pecorino cheese typology. Cheese making was performed applying the technology for “Crescenza” cheese typology with some modifications. In particular, pasteurized ewes’ milk was inoculated with two commercial starter formulations (SF1 and SF2) of Streptococcus thermophilus to obtain two different productions (QdO-P1 and QdO-P2, respectively). Plate counts demonstrated the ability of both starter formulations to drive the fermentation process, since S. thermophilus counts reached 109 CFU/g in both productions. Generally, the two starter formulations did not affect the chemical composition of QdO cheeses that contained, on average, 64.08% dry matter of which approximately 54.99% were fats and 36.39% proteins. Among chemical parameters, significant differences were registered for secondary lipid oxidation state (significantly lower for QdO-P2), fatty acids and volatile organic compounds (VOCs). However, the differences registered among cheese VOCs from were not perceived by the panelists who recognized both cheese productions highly similar, although QdO-P2 cheeses were mostly appreciated by the judges. This study allowed to produce a novel fresh ovine cheese with specific chemical and sensorial characteristics well appreciated by consumers