Search

The increasing request for classic foods has led to a higher demand for cheeses enjoying a “recognition of quality” status. This phenomenon has also affected Sicilian dairy products, in particular protected designation of origin (PDO) “Vastedda della valle del Belìce” (VdB), a cheese traditionally available only during the summer season, but requested year round. The variations of the microbial populations of raw milk among the seasons influence the microbiological quality of the cheeses produced. This review article reports the historical importance of cheeses and the traditional equipment used for dairy productions in Sicily with a specific focus on VdB. The microbial ecology of this cheese is thoroughly discussed. The recent findings to stabilize the traditional VdB production protocol with regards to the microbial composition and the organoleptic characteristics of the final cheeses are reviewed. The selection of starter lactic acid bacteria from PDO cheeses, their in vitro and in vivo application, and, overall, the setup of their inoculation, have all generated a novel production protocol. The innovation was respectful of the traditional discipline, essentially based on the ad hoc biofilm formation on the traditional wooden equipment, and allowed the preservation of VdB typicality. Future prospects are summarized.

Background: The continuous worldwide increase of consumption of fresh mushrooms has registered in the recent years. The major goal of this study was to determine the microbiological characteristics of wild edible mushrooms and effect of temperature during storage of Morchella conica. Methods: Wild mushrooms of the species Boletus edulis, Cantharellus cibarius, and Leccinum aurantiacum were collected in a mixed forest of Picea abies, Betula pendula, and Pinus sylvestris located in Tartu territory, Estonia. Faecal indicators, potential pathogens, spoilage bacteria, and microfungi (yeasts and moulds) were evaluated. M. conica was microbiologically investigated for 24 days under different thermal regimes, including 4, 8, 12, 15, 20, and 28 °C. The statistical analysis was conducted with SAS 9.2 software. Results: The microbial counts of wild mushrooms, ranging from 6.81 to 7.68 log10 CFU/g for total mesophilic count, were generally higher (p

The main aim of this work was to investigate on kefir grains (KGs) increase using different dairy by-products. To this purpose, whey and deproteinized whey from cow, goat and sheep milk were compared to the pasteurized milk of the corresponding animal species. Each substrate was inoculated with 3% (w/v) of KGs cultivated in ultra-high temperature cow milk and evalu- ated for pH decrease, total titratable acidity increase and development of lactic acid bacteria (LAB) and yeasts released in the matrices after 24 h incubation at three different temperatures (20, 25 and 30 °C). The genotypic characterization of the dominating microbial populations resulted in the identification of two LAB belonging to the species Lactococcus lactis and Leuconostoc mesenteroides and one yeast strain within the species Kluyveromyces marxianus. Among whey based fermenta- tion media deproteinized sheep whey allowed the highest KGs increase (almost 50%). Results of the present investigations reveal the potential of the dairy wastes for KGs production at industrial level.

Early vat bacterial biofilms developed spontaneously through contact with whey have been characterized on seven wood types (Castanea sativa Miller, Cedrus libani, A. Rich., Prunus avium L., Fraxinus ornus L., Juglans regia L., Pinus nigra J.F. Arnold and Populus nigra L.). The present study aimed to evaluate the influence of these biofilms on the microbiological, chemical, physical and sensory characteristics of PDO Vastedda della valle del Belìce (VdB) cheese, processed traditionally from raw ewe's milk using wooden tools. To this purpose, the experimental cheeses after 15 d of refrigerated storage were examined. Lactic acid bacteria (LAB) populations dominated the microbial community of all samples. The species more frequently identified were Lactococcus lactis among starter LAB and Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum and Pediococcus pentosaceus among non starter LAB. Culture-independent analysis of microbiota diversity was performed by MiSeq Illumina that identified Streptococcus as major group followed by members of Enterobacteriaceae family, Lactococcus and Lactobacillus. Generally, the seven tree species did not negatively affect the physicochemical composition of VdB cheeses. Chestnut (both Sicilian and Calabrian) vats produced cheeses with significant lower hue angle (a*/b*) than other wood types. Among chemical parameters, significant variations were registered for a w , primary and secondary lipid oxidation state (significantly lower for the VdB cheeses produced with poplar wood), and volatile organic compounds (VOCs). The significant differences detected among the VOCs emitted from cheeses were not perceived by the panelists who recognized all cheeses from the different trials as similar. This study confirmed the suitability of cedar, cherry, ash, walnut, black pine and poplar as alternative woods to chestnut for the production of the wooden vats employed in cheese making for the Sicilian traditional dairy productions.

The main hypothesis of this work was that Sicilian forestry resources are suitable for the production of equipment to be used in cheese making and indigenous milk lactic acid bacteria (LAB) are able to develop stable biofilms providing starter and nonstarter cultures necessary for curd fermentation and cheese ripening, respectively. Hence, the present work was carried out with deproteinized whey to evaluate LAB biofilm formation on different woods derived from tree species grown in Sicily. Microbiological and scanning electron microscopy analyses showed minimal differences in microbial levels and compositions for the neoformed biofilms. The specific investigation of Salmonella spp., Listeria monocytogenes , Escherichia coli , coagulase-positive staphylococci (CPS), and sulfite-reducing anaerobes did not generate any colony for all vats before and after bacterial adhesion. LAB populations dominated all vat surfaces. The highest levels (7.63 log CFU/cm 2 ) were registered for thermophilic cocci. Different colonies were characterized physiologically, biochemically, and genetically (at strain and species levels). Six species within the genera Enterococcus , Lactobacillus , Lactococcus , and Streptococcus were identified. The species most frequently present were Lactobacillus fermentum and Lactococcus lactis . LAB found on the surfaces of the wooden vats in this study showed interesting characteristics important for dairy manufacture. To thoroughly investigate the safety of the wooden vat, a test of artificial contamination on new Calabrian chestnut (control wood) vats was carried out. The results showed that LAB represent efficient barriers to the adhesion of the main dairy pathogens, probably due to their acidity and bacteriocin generation. IMPORTANCE This study highlights the importance of using wooden vats for traditional cheese production and provides evidence for the valorization of the Sicilian forest wood resources via the production of dairy equipment.

The survival of eight undesired (spoilage/pathogenic) food related bacteria (Citrobacter freundii PSS60, Enterobacter spp. PSS11, Escherichia coli PSS2, Klebsiella oxytoca PSS82, Serratia grimesii PSS72, Pseudomonas putida PSS21, Stenotrophomonas maltophilia PSS52 and Listeria monocytogenes ATCC 19114T) was investigated in mineral nutrient solution (MNS) during the crop cycle of radishes (Raphanus sativus L.) cultivated in hydroponics in a greenhouse. MNSs were microbiologically analyzed weekly by plate count. The evolution of the pure cultures was also evaluated in sterile MNS in test tubes. The inoculated trials contained an initial total mesophilic count (TMC) ranging between 6.69 and 7.78 Log CFU/mL, while non-sterile and sterile control trials showed levels of 4.39 and 0.97 Log CFU/mL, respectively. In general, all inoculated trials showed similar levels of TMC in MNS during the experimentation, even though the levels of the inoculated bacteria decreased. The presence of the inoculums was ascertained by randomly amplified polymorphic DNA (RAPD) analysis applied on the isolates collected at 7-day intervals. At harvest, MNSs were also analyzed by denaturing gradient gel electrophoresis (DGGE). The last analysis, except P. putida PSS21 in the corresponding trial, did not detect the other bacteria, but confirmed that pseudomonads were present in un-inoculated MNSs. Despite the high counts detected (6.44 and 7.24 CFU/g), only C. freundii PSS60, Enterobacter spp. PSS11 and K. oxytoca PSS82 were detected in radishes in a living form, suggesting their internalization.

Three Lactococcus lactis subsp. cremoris strains were used to develop ad hoc biofilms on the surfaces of virgin wooden vats used for cheese production. Two vats (TZ) were tested under controlled conditions (pilot plant), and two vats (TA) were tested under uncontrolled conditions (industrial plant). In each plant, one vat (TA1 and TZ1) was used for the control, traditional production of PDO Vastedda della Valle del Belìce (Vastedda) cheese, and one (TA2 and TZ2) was used for experimental production performed after lactococcal biofilm activation and the daily addition of a natural whey starter culture (NWSC). Microbiological and scanning electron microscopy analyses showed differences in terms of microbial levels and composition of the neoformed biofilms. The levels of the microbial groups investigated during cheese production showed significant differences between the control trials and between the control and experimental trials, but the differences were not particularly marked between the TA2 and TZ2 productions, which showed the largest numbers of mesophilic lactic acid bacterium (LAB) cocci. LAB populations were characterized phenotypically and genotypically, and 44 dominant strains belonging to 10 species were identified. Direct comparison of the polymorphic profiles of the LAB collected during cheese making showed that the addition of the NWSC reduced their biodiversity. Sensory evaluation showed that the microbial activation of the wooden vats with the multistrain Lactococcus culture generated cheeses with sensory attributes comparable to those of commercial cheese. Thus, neoformed biofilms enable a reduction of microbial variability and stabilize the sensorial attributes of Vastedda cheese.

Traditional cheeses made without starter cultures can be characterised by the attribute of instability. The addition of autochthonous starter cultures can ensure stability without compromising the characteristics of the final product. This study aimed to characterise the autochthonous lactic acid bacteria (LAB) population in “Vastedda della valle del Belice” cheeses, which have a protected designation of origin (PDO) status, in order to develop an ad hoc starter culture to be used in its future production. Winter and spring productions were analysed to ensure isolation of specific LAB that had adapted to perform fermentation at low temperatures. Plate counts revealed total microbial numbers nearing 109 CFU.g−1. All of the cheese samples were dominated by coccus-shaped LAB. When enterobacteria were present, their concentrations were at similar levels (3.3–5.6 Log CFU.g−1) in both seasons. All of the colonies that differed in morphological appearance were isolated and differentiated on the basis of phenotypic characteristics and genetic polymorphisms, as analysed by random amplification of polymorphic DNA-polymerase chain reaction. A total of 74 strains were identified and further genotyped by sequencing the 16S rRNA gene, resulting in the identification of 16 LAB species belonging to five genera (Enterococcus, Lactobacillus, Lactococcus, Leuconostoc and Streptococcus). The species most frequently present were Streptococcus gallolyticus subsp. macedonicus, Streptococcus thermophilus, Lactococcus lactis and Leuconostoc mesenteroides. The 74 strains were also investigated in vitro for general dairy parameters such as acidification capacity, diacetyl generation and antibacterial activity. Several strains of the most frequently represented species displayed traits relevant to the production of PDO “Vastedda della valle del Belice”.

The microbial composition of artisan and industrial animal rennet pastes was studied by using both culture-dependent and -independent approaches. Pyrosequencing targeting the 16S rRNA gene allowed to identify 361 operational taxonomic units (OTUs) to the genus/species level. Among lactic acid bacteria (LAB), Streptococcus thermophilus and some lactobacilli, mainly Lactobacillus crispatus and Lactobacillus reuteri , were the most abundant species, with differences among the samples. Twelve groups of microorganisms were targeted by viable plate counts revealing a dominance of mesophilic cocci. All rennets were able to acidify ultrahigh-temperature-processed (UHT) milk as shown by pH and total titratable acidity (TTA). Presumptive LAB isolated at the highest dilutions of acidified milks were phenotypically characterized, grouped, differentiated at the strain level by randomly amplified polymorphic DNA (RAPD)-PCR analysis, and subjected to 16S rRNA gene sequencing. Only 18 strains were clearly identified at the species level, as Enterococcus casseliflavus , Enterococcus faecium , Enterococcus faecalis , Enterococcus lactis , Lactobacillus delbrueckii , and Streptococcus thermophilus , while the other strains, all belonging to the genus Enterococcus , could not be allotted into any previously described species. The phylogenetic analysis showed that these strains might represent different unknown species. All strains were evaluated for their dairy technological performances. All isolates produced diacetyl, and 10 of them produced a rapid pH drop in milk, but only 3 isolates were also autolytic. This work showed that animal rennet pastes can be sources of LAB, mainly enterococci, that might contribute to the microbial diversity associated with dairy productions.

In this study, the composition of the cultivable microbial populations of 38 nectar honey and honeydew honey samples of different botanical and geographical origin were assessed. After growth in specific media, various colonies with different appearance were isolated and purified before phenotypic (morphological, physiological and biochemical traits) and genotypic [randomly amplified polymorphic DNA (RAPD), repetitive DNA elements-PCR (rep-PCR) and restriction fragment length polymorphism (RFLP)] differentiation. The identification was carried out by 16S rRNA gene sequencing for bacteria and, in addition to RFLP, by sequencing the D1/D2 region of the 26S rRNA gene for yeasts and the 5.8S-ITS rRNA region for filamentous fungi. The results showed the presence of 13 species of bacteria, 5 of yeasts and 17 of filamentous fungi; the species most frequently isolated were Bacillus amyloliquefaciens, Zygosaccharomyces mellis and Aspergillus niger for the three microbial groups, respectively. The highest microbial diversity was found in multifloral honeys. No correlation among the microbial species and the botanical/geographical origin was found, but some strains were highly adapted to these matrices since they were found in several samples of different origin.

article Twelve lactic acid bacteria (LAB), previously selected in vitro (Gaglio et al., 2014), were evaluated in situ for their potential to act as starter cultures for the continuous four-season production of Vastedda-like cheese, made with raw ewes' milk. The strains belonged to Lactobacillus delbrueckii, Lactococcus lactis subsp. cremoris, Leuconostoc mesenteroides subsp. mesenteroides and Streptococcus thermophilus .L AB werefirst inoculated in multiple-strain combinations on the basis of their optimal growth temperatures in three process conditions which differed for milk treatment and medium for strain development: process 1, growth of strains in the optimal synthetic media and pasteurised milk; process 2, growth of strains in whey based medium (WBM) and pasteurised milk; and process 3, growth of strains in WBM and raw milk. The strains that acidified the curds in short time, as shown by a pH drop, were all mesophilic and were then tested in a single inoculum through process 3. Ran- domly amplified polymorphic DNA (RAPD)-PCR analysis applied to the colonies isolated from the highest dilu- tions of samples confirmed the dominance of the added strains after curd acidification, stretching and storage. After 15 days of refrigerated storage, the decrease in pH values showed an activity of the mesophilic strains at low temperatures, but only Lc. lactis subsp. cremoris PON153, Ln. mesenteroides subsp. mesenteroides PON259 and PON559 increased their number during the 15 days at 7 °C. A sensory evaluation indicated that the cheeses obtained by applying protocol 3 and by inoculation with lactococci are the most similar to the protected denom- ination oforigin(PDO) cheese andreceivedthebest scores bythe judges. Thus,theexperimentalcheesesobtain- ed with raw milk and inoculated with single and multiple combinations of lactococci were subjected to the analysis of the volatile organic compounds (VOCs) carried out by a headspace solid phase microextraction (SPME) technique coupled with gas chromatography with mass spectrometric detection (GC/MS). The domi- nance of lactococci over thermophilic LAB of raw milk was verified during summer production and, based on the combination of VOC profiles and sensory evaluation of the final cheeses, the multi-strain Lactococcus culture resulted in the most suitable starter preparation for the full-year production of Vastedda-like cheese.

The composition of yeast and lactic acid bacteria (LAB) communities and the chemical evolution of the large-scale commercial vinification of Catarratto IGT Sicilia, carried out under the biological regime, was followed from grape harvest until bottling. Simultaneously to the maximum growth of yeasts, LAB counts reached high level of concentration (6-7 log CFU mL(-1)) during the first steps of the alcoholic fermentation. Yeast identification was determined applying different molecular methods. The highest species biodiversity was observed on grape and must samples taken soon after pressing. Saccharomyces cerevisiae was detected at dominant concentrations during the entire winemaking process. LAB cultures were grouped and identified by a combined phenotypic and genotypic approach. Leuconostoc mesenteroides, Lactobacillus hilgardii and Lactobacillus plantarum species were identified; the last was the main LAB recognized during vinification. The winemaking process was also chemically monitored. The alcoholic content was approximately 12.67% (v v(-1)) at bottling; pH, volatile acidity and total acidity showed a moderate increase during vinification. Tartaric, citric and malic acids decreased until bottling, while lactic acid showed a rapid increase at the end of maceration and bottling. Trans-caffeil tartaric acid was the most abundant phenolic compound and volatile organic compounds (VOC) were mainly represented by isoamylic alcohol, isobutanol, ethyl acetate and octanoic acid.