Microbial ecology of retail ready-to-eat escarole and red chicory sold in Palermo City, Italy

Background: Ready-To-Eat (RTE) foods include any edible food that is commonly consumed raw. This study aimed at evaluation of microbial ecology of retail RTE escarole and red chicory sold in Palermo city, Italy. Methods: A total of 32 mono-varietal RTE samples, including escarole (n=16) and red chicory (n=16) samples were obtained from Palermo, Italy. Both RTE vegetables at expiry date were analyzed to quantify spoilage bacteria, pathogenic bacteria, and yeast. All different colonies were isolated and identified on the basis of phenotypic characteristics and genetic polymorphisms by random amplification of polymorphic DNA-Polymerase Chain Reaction (PCR) and further genotype by sequencing the 16S rRNA gene. The statistical analysis was conducted with SAS 9.2 software (Statistical Analysis System Institute Inc., Cary, NC, USA). Results: The level of Listeria monocytogenes and coagulase-positive staphylococci were below the detection. Total microbial counts were above 8 log10 colony forming unit/g in RTE red chicory, while they were about 1 log cycle lower in escarole. In general, escarole showed lower levels for all microbial groups than red chicory with the exception of the total yeast. A total of 13 strains were identified into ten species belonging to six genera as Bacillus, Erwinia, Pantoea, Pseudomonas, Microbacterium, and Rahnella. The most numerous identified genera were Pseudomonas and Pantoea. Conclusion: This work pointed out the relevance of implementing good hygiene practices during processing in order to prolong quality parameters and acceptability of mono-varietal salads

Aloe-based edible coating to maintain quality of fresh-cut italian pears (Pyrus communis L.) during cold storage

Pear fruits are known for their antioxidant and nutritional characteristics. However, they are very susceptible to rapid decay. Edible coating (EC) represents a good strategy to maintain postharvest quality. The effects of two EC in slowing down the senescence processes in fresh-cut ‘Coscia’ pears were investigated: EC1 (A. vera gel, hydroxypropyl-methylcellulose and pomegranate seeds oil (PSO), EC2 (A. vera gel and hydroxypropyl-methylcellulose). Weight loss, firmness and colour decrease more slowly in both EC-treated than in untreated (CTR) slices; soluble solid content increases faster in CTR, indicating a faster ripening process. The specific investigation of undesired microorganisms did not generate any colony in all analysed samples. Sensory analysis confirmed that the tasters preferred the EC2-treated samples, as they were the only ones that did not show undesirable flavours until the last day of storage

Effect of Glucose and Inactivated Yeast Additions on the Fermentation Performances of Lactiplantibacillus pentosus OM13 during the Production of Nocellara del Belice Table Olives

The use of selected strains of lactic acid bacteria is necessary to produce fermented table olives with high hygiene and quality standards at the industrial level. A current tendency is the use of fermentation adjuvants (nutrients and activators) that can satisfy the nutritional needs of starter strains. In this study, five experimental protocols, different for nutrient and activator presence and addition of Lactiplantibacillus pentosus OM13 in freeze-dried form and after acclimatisation, were tested with the aim of improving the fermentation performances of the commercial starter. The trial inoculated with the starter strain acclimatised in the presence of nutrients and activator showed the most rapid acidification during the first phase of fermentation (third to ninth day), registering a pH loss of 3.40 units. The addition of adjuvants positively influences starter dominance (>89%) and rapid colonisation (>7 Log CFU/mL from third d) by indirectly limiting the presence of undesirable microorganisms. The analysis of volatile organic compounds revealed the presence of 32 chemicals distributed differently in each trial. Sensory evaluation showed that table olives produced with the different treatments were characterised by low bitterness, acidity, and absence of unpleasant odours/flavours. Control production showed slower acidification kinetics and lower sensory pleasantness than the other trials

Behavior of four main dairy pathogenic bacteria during manufacturing and ripening of pecorino siciliano cheese

Background: Consumption of raw cheese may be associated with different diseases. This study aimed to evaluate behavior of four pathogenic bacteria during manufacture and ripening of Protected Designation of Origin (PDO) Pecorino Siciliano cheese. Methods: The experimental cheese groups were inoculated with pathogenic bacteria, including Escherichia coli O157, Listeria monocytogenes, Salmonella Enteritidis, and Staphylococcus aureus. The cheese making processes were monitored from milk curdling until 3 months ripened cheeses and the levels of Lactic Acid Bacteria (LAB) and the four dairy pathogens were evaluated by plate counts. Randomly Amplified Polymorphic DNA (RAPD)-Polymerase Chain Reaction (PCR) analysis was applied to confirm that the colonies isolated during the several steps of production were the same strains added in milk. Statistical analysis was done using XLStat software. Results: The levels of mesophilic and thermophilic coccus and rod LAB in curd were comparable in both trials and reached values between 8-9 log10 Colony Forming Unit (CFU)/g in cheeses at 90 days of ripening. The four pathogenic bacteria were found in experimental curd at levels higher than those inoculated in milk and completely disappeared after 60 days of ripening. The RAPD analysis clearly demonstrated the presence of the added strain during production and confirmed the results of plate counts. Conclusion: This work showed that the production conditions of PDO Pecorino Siciliano cheese decreased growth of E. coli O157, L. monocytogenes, S. Enteritidis, and S. aureus

Application of hydrogen peroxide to improve the microbiological stability of food ice produced in industrial facilities

This work was aimed to produce an “active” food ice to preserve its microbiological safety over time. With this in mind, ice cubes were processed with the addition of H2O2 to water before freezing. Four food ice productions were performed at the industrial level: one control trial without the addition of H2O2 (0OX) and three experimental trials obtained by adding 4, 8, and 12 mg/L of H2 O2 (4OX, 8OX, and 12OX), respectively. After production, all food ice trials were artificially contaminated with 102 CFU/100 mL of water-borne pathogenic bacteria (Escherichia coli ATCC 25922, Enteroccus faecalis ATCC 29212, and Pseudomonas aeruginosa ATCC 27853) inoculated individually. Thawed ice samples were then subjected to microbiological analyses performed by the membrane filtration method and the results indicated that only trial 12OX was able to inactivate all bacteria strains. In conclusion, the addition of 12 mg/L H2O2 represents an optimal cost-effective strategy to preserve the microbiological stability of food ice even when it is improperly handled after production

The influence of the wooden equipment employed for cheese manufacture on the characteristics of a traditional stretched cheese during ripening

The influence of the wooden equipment used for the traditional cheese manufacturing from raw milk was evaluated on the variations of chemico-physical characteristics and microbial populations during the ripening of Caciocavallo Palermitano cheese. Milk from two farms (A, extensive; B, intensive) was processed in traditional and standard conditions. Chemical and physical traits of cheeses were affected by the farming system and the cheese making technology, and changed during ripening. Content in NaCl and N soluble was lower, and paste consistency higher in cheese from the extensive farm and traditional technology, whereas ripening increased the N soluble and the paste yellow and consistency. The ripening time decreased the number of all lactic acid bacteria (LAB) groups, except enterococci detected at approximately constant levels (104 and 105 cfu g-1 for standard and traditional cheeses, respectively), till 120 d of ripening. In all productions, at each ripening time, the levels detected for enterococci were lower than those for the other LAB groups. The canonical discriminant analysis of chemical, physical and microbiological data was able to separate cheeses from different productions and ripening time. The dominant LAB were isolated, phenotypically characterised and grouped, genetically differentiated at strain level and identified. Ten species of LAB were found and the strains detected at the highest levels were Pediococcus acidilactici and Lactobacillus casei. Ten strains, mainly belonging to Lactobacillus rhamnosus and Lactobacillus fermentum showed an antibacterial activity. The comparison of the polymorphic profiles of the LAB strains isolated from the wooden vat with those of the strains collected during maturation, showed the persistence of three enterococci in traditional cheeses, with E. faecalis found at dominant levels over the Enterococcus population till 120 d; the absence of these strains in the standard productions evidenced the contribution of vat LAB during Caciocavallo Palermitano cheese ripening

Tracking the transfer of antimicrobial resistance genes from raw materials to sourdough breads

The present study hypothesizes that raw materials used in bread making can transfer antibiotic resistance genes (ARGs) to processed breads. Four types of flour and four types of semolina were purchased from supermarkets and inoculated with a commercial dried sourdough starter to make breads. The microbiological characteristics of all raw materials and fermented doughs were investigated. The levels of yeasts and lactic acid bacteria (LAB) increased up to 107 CFU/g. The values of pH decreased to 4.54–4.86 while total titratable acidity increased inversely. All unprocessed and processed samples, including breads, were analyzed by a molecular approach to detect bacterial and fungal DNAs and 17 antibiotic resistance genes for penicillins, macrolides, tetracyclines, and chloramphenicol. Illumina technology showed that the operational taxonomy units (OTUs) identified from unprocessed wheat milling products, fermented doughs, and baked products mainly belonged to Acetobacteraceae. Enterococci were present in all doughs. After baking, the relative abundance (RA)% of Enterococcus and Acetobacteraceae decreased. The DNA analyzed for fungal composition showed that Kazachstania humilis dominated dried sourdough starter and doughs, and its OTUs were also detected at high RA% in baked products. The search for ARGs revealed that all samples analyzed did not show resistance to penicillins, chloramphenicol, and macrolides. However, three of the semolinas included in this study (S1, S3 and S4) and the corresponding doughs (SD1, SD3 and SD4) were positive for tet(A) and tet(B) resistance genes. This work indicated that breads have a limited role in the dissemination of ARG

Tracking the transfer of antimicrobial resistance genes from raw materials to sourdough breads

The present study hypothesizes that raw materials used in bread making can transfer antibiotic resistance genes (ARGs) to processed breads. Four types of flour and four types of semolina were purchased from supermarkets and inoculated with a commercial dried sourdough starter to make breads. The microbiological characteristics of all raw materials and fermented doughs were investigated. The levels of yeasts and lactic acid bacteria (LAB) increased up to 107 CFU/g. The values of pH decreased to 4.54–4.86 while total titratable acidity increased inversely. All unprocessed and processed samples, including breads, were analyzed by a molecular approach to detect bacterial and fungal DNAs and 17 antibiotic resistance genes for penicillins, macrolides, tetracyclines, and chloramphenicol. Illumina technology showed that the operational taxonomy units (OTUs) identified from unprocessed wheat milling products, fermented doughs, and baked products mainly belonged to Acetobacteraceae. Enterococci were present in all doughs. After baking, the relative abundance (RA)% of Enterococcus and Acetobacteraceae decreased. The DNA analyzed for fungal composition showed that Kazachstania humilis dominated dried sourdough starter and doughs, and its OTUs were also detected at high RA% in baked products. The search for ARGs revealed that all samples analyzed did not show resistance to penicillins, chloramphenicol, and macrolides. However, three of the semolinas included in this study (S1, S3 and S4) and the corresponding doughs (SD1, SD3 and SD4) were positive for tet(A) and tet(B) resistance genes. This work indicated that breads have a limited role in the dissemination of ARGs

In vivo application and dynamics of lactic acid bacteria for the four-season production of Vastedda-like cheese.

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, madewith raw ewes' milk. The strains belonged to Lactobacillus delbrueckii, Lactococcus lactis subsp. cremoris, Leuconostoc mesenteroides subsp. mesenteroides and Streptococcus thermophilus. LABwere first 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. Randomly amplified polymorphic DNA (RAPD)-PCR analysis applied to the colonies isolated from the highest dilutions 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 \ub0C. A sensory evaluation indicated that the cheeses obtained by applying protocol 3 and by inoculation with lactococci are the most similar to the protected denomination of origin (PDO) cheese and received the best scores by the judges. Thus, the experimental cheeses obtained 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 dominance 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

Monitoring commercial starter culture development in presence of red grape pomace powder to produce polyphenol-enriched fresh ovine cheeses at industrial scale level

Red grape Nero d'Avola cultivar grape pomace powder (GPP) was applied during fresh ovine cheese production in order to increase polyphenol content. Before cheeses were produced, the bacteria of a freeze-dried commercial starter culture were isolated and tested in vitro against GPP. Two dominant strains, both resistant to GPP, were identified. Thestarter culture was inoculated in pasteurized ewe's milk and the curd was divided into two bulks, one added with 1% (w/w) GPP and another one GPP-free. GPP did not influence the starter culture development, since lactic acid bacteria (LAB) counts were 109 CFU/g in both cheeses at 30 d. To exclude the interference of indigenous LAB, the pasteurized milk was analyzed, and several colonies of presumptive LAB were isolated, purified and typed. Four strains were allotted into Enterococcus and Lacticaseibacillus genera. The direct comparison of the polymorphic profiles of cheese bacteria evidenced the dominance of the starter culture over milk LAB. The addition of GPP increased cheese total phenolic compounds by 0.42 g GAE/kg. Sensory evaluation indicated that GPP-enriched cheese was well appreciated by the judges, providing evidence that GPP is a suitable substrate to increase the availability of total phenolic content in fresh ovine cheese