Lacto-fermented garlic handcrafted in the Lower Silesia Region (Poland): Microbial diversity, morpho-textural traits, and volatile compounds.

The aim of the present study was to provide a first characterization of lacto-fermented garlic manufactured by local small-scale artisanal producers in the Lower Silesia Region (Poland). The lacto-fermented garlic samples showed high nutritional features in terms of antioxidant activity. A total of 86 compounds, belonging to various chemical classes, were identified by headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC/MS). Most of these compounds belonged to six main classes, being sulfur compounds, esters and acetates, oxygenated monoterpenes, monoterpene hydrocarbons, and alcohols. Aldehydes, acids, ketones, furans, and phenols were also identified. In the analyzed samples, counts up to 8 log cfu g-1 were observed for lactic acid bacteria. Metataxonomic analysis revealed the presence of Levilactobacillus, Lactiplantibacillus, Latilactobacillus, Secundilactobacillus, Weissella, Leuconostoc, Lactococcus, Pediococcus, and Lacticaseibacillus among the major taxa. These results were confirmed by the isolation and characterization of viable lactic acid bacteria. Indeed, the presence of the closest relatives to Lacticaseibacillus casei group, Pediococcus parvulus, Levilactobacillus brevis, Levilactobacillus parabrevis, and Lactiplantibacillus plantarum group was observed. A good acidification performance in salty garlic-based medium was observed for all the isolates that, between 8 and 15 days of fermentation, reached pH values comprised between 4 and 3.5, depending on the tested species. Of note, 15 out of the 37 lactic acid bacteria isolates (Levilactobacillus parabrevis, Pediococcus parvulus, Lactiplantibacillus plantarum group, and Lacticaseibacillus casei group) showed the presence of the hdcA gene of Gram-positive bacteria encoding for histidine decarboxylase. Furthermore, for 8 out of the 37 isolates the in-vitro exopolysaccharides production was observed. No isolate showed inhibitory activity against the three Listeria innocua strains used as surrogate for Listeria monocytogenes.

Technological and Enzymatic Characterization of Autochthonous Lactic Acid Bacteria Isolated from Viili Natural Starters.

Viili, a Finnish ropy fermented milk, is traditionally manufactured through spontaneous fermentation, by mesophilic lactic acid bacteria and yeast-like fungi, or back-slopping. This study evaluated four natural viili starters as sources of lactic acid bacteria for dairy production. Back-slopping activation of the studied viili samples was monitored through pH and titratable acidity measurements and enumeration of mesophilic lactic acid bacteria. Sixty lactic acid bacteria isolates were collected, molecularly identified, and assayed for acidification performance, enzymatic activities, production of exopolysaccharides (EPSs), presence of the histidine decarboxylase (hdcA) gene of Gram-positive bacteria, and production of bacteriocins. A neat predominance of Lactococcus lactis emerged among the isolates, followed by Enterococcus faecalis, Enterococcus faecium, Enterococcus durans, Enterococcus lactis, and Lactococcus cremoris. Most isolates exhibited proteolytic activity, whereas only a few enterococci showed lipase activity. Five isolates identified as L. cremoris, L. lactis, and E. faecalis showed a good acidification performance. Most of the isolates tested positive for leucine arylamidase, whereas only one E. durans and two L. lactis isolates were positive for valine arylamidase. A few isolates also showed a positive reaction for beta-galactosidase and alpha- and beta-glucosidase. None of the isolates produced EPSs or bacteriocins. The hdcA gene was detected in five isolates identified as L. lactis and E. faecium. A few L. cremoris and L. lactis isolates for potential use as starter or adjunct cultures for dairy processing were finally identified.

Exploitation of Black Olive (Olea europaea L. cv. Piantone di Mogliano) Pomace for the Production of High-Value Bread.

In this study, the morpho-textural features, total phenolic content (TPC), and antioxidant capacity (AOC) of bread fortified with olive (Olea europaea L.) pomace were evaluated. Fresh olive pomace was subjected to microbiological and chemical (TPC, AOC, and fiber) analyses; then, the same olive pomace was analyzed during 1 to 6 months of storage at 4 °C or -20 °C. All olive pomace samples were used in 10%, 15%, or 20% amounts to produce type 0 soft wheat (Triticum aestivum) and whole wheat bread samples. The volatile organic compounds (VOCs) in the bread samples were also analyzed to assess the effect of the addition of the olive pomace on the flavor profile of the baked products. The TPC and AOC evaluation of olive pomace showed no differences among the analyzed samples (fresh, refrigerated, or frozen). Regarding the bread containing olive pomace, the specific volume was not affected by the amount or the storage methods of the added pomace. Bread samples produced with soft wheat flour showed the lowest hardness values relative to those produced with whole wheat flour, irrespective of the amount or storage method of the olive pomace. Regarding color, the crust and crumb of the bread samples containing 20% olive pomace were significantly darker. The bread samples containing 20% olive pomace had the highest TPC. The bread samples with fresh olive pomace were characterized by terpenoids, ketones, and aldehydes, whereas the bread samples containing refrigerated olive pomace were characterized by alcohols (mainly ethanol), acids, esters, and acetate. Finally, the bread samples with frozen olive pomace showed a volatile profile similar to that of bread produced with fresh olive pomace. Olive pomace was shown to be a suitable ingredient for producing bread with high nutritional value.

Tasting of traditional Polish fermented cucumbers: Microbiology, morpho-textural features, and volatilome.

In the present study, naturally fermented and unpasteurized cucumbers (Cucumis sativus L.) collected from 4 producers located in different regions of Poland were studied. The fermented cucumbers were characterized by significant nutritional features in terms of polyphenols content and antioxidant activity. Microbiological analyses revealed active bacterial populations of lactococci, thermophilic cocci, lactobacilli, and coagulase-negative cocci. The microbiological characterization of cucumber and brine samples through metataxonomic analysis allowed the dominant species to be detected, being Lactococcus and Streptococcus in cucumbers, and Lactiplantibacillus, Leuconostoc, Pediococcus, Secundilactobacillus, and Lentilactobacillus in brine. The isolation activity offered a clear picture of the main active lactic acid bacteria at the end of fermentation, being Pediococcus parvulus and Lactiplantibacillus plantarum group. All the studied isolates showed a good attitude in fermenting a cucumber-based broth, thus suggesting their potential application as starter or adjunct cultures for guided cucumber fermentation. Moreover, for the same isolates, strong aminopeptidase activity (due to leucine arylamidase and valine arylamidase) was observed, with potential effect on the definition of the final sensory traits of the product. Only a few isolates showed the ability to produce exopolysaccharides in synthetic medium. Of note, the presence of the hdcA gene in some Pediococcus ethanolidurans isolates also confirmed the need for a thorough characterization of starter candidates to avoid undesired adverse effects on consumer's health. No isolate showed the production of bacteriocins against Listeria innocua used as surrogate for Listeria monocytogenes. Based on the results of Headspace Solid-Phase Microextraction-Gas Chromatography/Mass Spectrometry analysis, a rich and complex volatilome, composed by more than 80 VOCs, was recognized and characterized. In more detail, the detected compounds belonged to 9 main classes, being oxygenated terpenes, alcohols, terpenes, ketones, acids, aldehydes, esters, sulfur, and sesquiterpenes.

Search for carbapenem-resistant bacteria and carbapenem resistance genes along swine food chains in Central Italy.

The presence of carbapenem-resistant bacteria and carbapenem resistance genes (CRGs) in livestock is increasing. To evaluate the presence of carbapenemase-producing Enterobacteriaceae (CPE) and the main CRGs along swine food chains of the Marche Region (Central Italy), samples of faeces, feed, and animal-food derived products were collected from seven small/medium, medium, and large-scale pig farms. A total of 191 samples were analysed using a culture-dependent method, with the aim of isolating CPE. Isolates were analysed for their resistance to carbapenems using a modified Hodge test and the microdilution method for the minimum inhibitory concentration (MIC) determination. Moreover, the extraction of microbial DNA from each sample was performed to directly detect selected CRGs via qPCR. Among the 164 presumptive resistant isolates, only one strain from a liver sample, identified as Aeromonas veronii, had an ertapenem MIC of 256 µg/mL and carried a carbapenemase- (cphA) and a ß-lactamase- (blaOXA-12) encoding genes. A low incidence of CRGs was found; only nine and four faecal samples tested positive for blaNDM-1 and blaOXA-48, respectively. Overall, the importance of monitoring CPE and CRGs in livestock and their food chains should be stressed to control all potential non-human CPE and CRGs reservoirs and to determine safety levels for human health.

Spotlight on autochthonous microbiota, morpho-textural characteristics, and volatilome of a traditional Polish cold-smoked raw sausage.

The aim of the present study was to obtain information on the bacterial diversity of traditional Polish cold-smoked raw sausages (Kielbasa polska wedzona) manufactured by two artisanal producers using different selective growth media and a metataxonomic analysis. Physico-chemical and morpho-textural characteristics were also carried out, together with Microextraction-Gas Chromatography/Mass Spectrometry (HS-SPMEGC/MS) to study the volatile organic compounds (VOCs). The results overall obtained allowed a picture of the microbiota, the morpho-textural characteristics, and the volatilome of traditional Polish cold-smoked raw sausages (Kielbasa polska wedzona) to be drawn for the first time. In more detail, viable counting revealed active populations of presumptive lactobacilli, enterococci, coagulase-negative cocci, and a few spoilage microorganisms typically occurring in raw meat products. The metataxonomic analysis revealed the dominance of Latilactobacillus sakei occurring with a relative frequency between 77% and 89%. Pediococcus pentosaceus, Weissella hellenica, and Leuconostoc carnosum were detected among the minority taxa. In the sausages herein studied, no histamine levels of concern were detected. The Principal Component Analysis (PCA) performed on the Amplicon Sequence Variants (ASVs) did not show significant differences in the microbiota composition among producers. The HS-SPMEGC/MS analysis allowed the detection and identification of more than 90 volatile components belonging to ten main classes, namely: aldehydes, ketones, esters and acetates, acids, alcohols, phenols, furans, sulphur compounds, terpenoids, and benzene derivatives. The detected VOCs originated from spices, smoke, and microbial metabolism. The PCA of volatile compounds allowed differences between the sausage samples of the two producers to be identified.

Impact of Different Drying Methods on the Microbiota, Volatilome, Color, and Sensory Traits of Sea Fennel (Crithmum maritimum L.) Leaves.

Sea fennel (Crithmum maritimum L.) is a strongly aromatic herb of the Apiaceae family, whose full exploitation by the modern food industry is of growing interest. This study aimed at investigating the microbiological quality, volatile profile, and sensory traits of sea fennel spices produced using room-temperature drying, oven drying, microwave drying, and freeze drying. All the assayed methods were able to remove moisture up until water activity values below 0.6 were reached; however, except for microwave drying, none of the assayed methods were effective in reducing the loads of contaminating microorganisms. The metataxonomic analysis highlighted the presence of phytopathogens and even human pathogens, including members of the genera Bacillus, Pseudomonas, Alternaria, and Cryptococcus. When compared to fresh leaves, dried leaves showed increased L* (lightness) and c* (chroma, saturation) values and reduced hue angle. Dried leaves were also characterized by decreased levels of terpene hydrocarbons and increased levels of aldehydes, alcohols, and esters. For the sensory test, the microwave-dried samples obtained the highest appreciation by the trained panel. Overall, the collected data indicated microwave drying as the best option for producing sea fennel spices with low microbial loads, brilliant green color, and high-quality sensory traits.

Sea Fennel (Crithmum maritimum L.) as an Emerging Crop for the Manufacturing of Innovative Foods and Nutraceuticals.

Sea fennel (Crithmum maritimum L.) is a perennial, strongly aromatic herb that has been used since ancient times in cuisine and folk medicine due to its renowned properties. Recently described as a "cash" crop, sea fennel is an ideal candidate for the promotion of halophyte agriculture in the Mediterranean basin due to its acknowledged adaptation to the Mediterranean climate, its resilience to risks/shocks related to climate changes, and its exploitability in food and non-food applications, which generates an alternative source of employment in rural areas. The present review provides insight into the nutritional and functional traits of this new crop as well as its exploitation in innovative food and nutraceutical applications. Various previous studies have fully demonstrated the high biological and nutritional potential of sea fennel, highlighting its high content of bioactive compounds, including polyphenols, carotenoids, ω-3 and ω-6 essential fatty acids, minerals, vitamins, and essential oils. Moreover, in previous studies, this aromatic halophyte showed good potential for application in the manufacturing of high-value foods, including both fermented and unfermented preserves, sauces, powders, and spices, herbal infusions and decoctions, and even edible films, as well as nutraceuticals. Further research efforts are needed to fully disclose the potential of this halophyte in view of its full exploitation by the food and nutraceutical industries.

Identification of plasmids co-carrying cfr(D)/optrA and cfr(D2)/poxtA linezolid resistance genes in two Enterococcus avium isolates from swine brain.

Oxazolidinones are critically important antibiotics to treat human infections caused by multidrug-resistant bacteria, therefore the occurrence of linezolid-resistant enterococci from food-producing animals poses a serious risk to human health. In this study, Enterococcus avium 38157 and 44917 strains, isolated from the brain of two unrelated piglets, were found to carry the linezolid resistance genes cfr(D)-optrA, and cfr(D2)-poxtA, respectively. Whole genome sequencing analysis of E. avium 38157 revealed that the genes were co-located on the 36.5-kb pEa_cfr(D)-optrA plasmid showing high identity with the pAT02-c of Enterococcus faecium AT02 from pet food. The optrA region, was 99% identical to the one of the pAv-optrA plasmid from a bovine Aerococcus viridans strain, whereas the cfr(D) genetic context was identical to that of the plasmid 2 of E. faecium 15-307.1. pEa_cfr(D)-optrA was not transferable to enterococcal recipients. In E. avium 44917 a cfr(D)-like gene, named cfr(D2), and the poxtA gene were co-located on the transferable 42.6-kb pEa-cfr(D2)-poxtA plasmid 97% identical to the Tn6349 transposon of the human MRSA AOUC-0915. The cfr(D2) genetic context, fully replaced the Tn6644 that in S. aureus AOUC-0915 harbor the cfr gene. In conclusion, this is, the best of our knowledge, the first report of the new cfr(D2) gene variant. The occurrence of plasmids co-carrying two linezolid resistance genes in enterococci from food-producing animals needs close surveillance to prevent their spread to human pathogens.

Microbial Dynamics of a Specialty Italian Raw Ewe's Milk Cheese Curdled with Extracts from Spontaneous and Cultivated Onopordum tauricum Willd.

Milk coagulants prepared by maceration of flowers harvested from both spontaneous and cultivated Onopordum tauricum Willd. and a commercially available coagulant obtained from Cynara cardunculus L. (control) were assayed for small-scale manufacturing of Caciofiore, an Italian specialty raw ewe's milk cheese produced in a family run dairy farm located in the Marche region (Central Italy). The microbiota of the three thistle-based milk coagulants and their effect on the microbial dynamics of raw milk cheeses during fermentation and maturation (from day 0 up until day 60) were investigated through a combined approach based on viable counting and Illumina DNA sequencing. In both the control and experimental cheeses, despite the slight differences emerged depending on the coagulant used, Lactococcus lactis and Debaryomyces hansenii were the prevalent species among bacteria and fungi, respectively. Moreover, raw ewe's milk was the main factor affecting the evolution of both the bacterial and fungal microbiota in all cheeses. The overall similarities between control and experimental cheeses herein analyzed supports the exploitation of Onopordum tauricum Willd. as an alternative milk coagulating agent for production of Caciofiore and, more in general, raw ewe's milk cheeses.

Croatian white grape variety Marastina: First taste of its indigenous mycobiota.

The indigenous vineyard mycobiota contribute both to wine quality and vineyard sanitary status. Wines made from same grape variety but from different geographical locations are appreciated for their diversity. Because no information on indigenous mycobiota of Croatian grapevines is available, the aim of the present study was to start filling this knowledge gap by characterizing the indigenous mycobiota of Marastina variety. The use of metataxonomic approach has enabled the identification of 25 different fungal genera present on Marastina grape berries collected from 11 vineyards located within the Croatian coastal winegrowing region of Dalmatia (Northern Dalmatia, Dalmatian hinterland, Central and Southern Dalmatia). The substantial regional and local scale differences in their distribution were observed. Overall, Aureobasidium was the dominant genus followed by Cladosporium and Metschnikowia. Botrytis and Plenodomus were associated with the vineyards located in Central and Southern Dalmatia, whereas Pichia was associated with Northern Dalmatia vineyards. The largest abundance of Buckleyzyma, Cladosporium, Eremothecium, Fusarium, Papiliotrema, and Rhodotorula was observed in Dalmatian hinterland. Moreover, data suggested that climate conditions and soil type partially influenced the distribution of fungal communities. The local-scale differences emerged also for the physicochemical characteristics of fresh musts. The high malic acid content supported the development of Metschnikowia, and inhibited Fusarium growth, whereas a positive correlation between Erysiphe and pH values was observed. Sporobolomyces and Cystobasidium were negatively associated with high glucose concentration. The revealing of Marastina indigenous mycobiota provided information on the members of fungal community negatively influencing the grapevine sanitary status as well as those which could be employed in disease biocontrol.

Microbiological, morpho-textural, and volatile characterization of Portuguese Queijo de Nisa PDO cheese.

Queijo de Nisa PDO (Protected Designation of Origin) is a semi-hard cheese obtained from raw ewe's milk clotted with cardoon (Cynara cardunculus, L.) rennet. The aim of the present study was to characterize the bacterial communities naturally occurring in Queijo de Nisa PDO cheese samples through viable counting and metataxonomic analysis. Moreover, physico-chemical and morpho-textural analyses were also performed, together with the analysis of volatile organic compounds (VOCs) through solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS). In the analyzed samples, pH values were comprised between 4.84 and 5.74. As for lactic acid, the concentration varied between 0.83 and 2.10 g 100 g-1. Water activity varied between 0.942 and 0.960. Lightness of the samples ranged from 107.82 to 119.16, whereas hardness ranged between 34.45 N and 126.05 N. As for microbiological analyses, lactococci and lactobacilli counts were up to 9.01 Log CFU g-1. Coagulase-negative cocci and enterococci counts were up to 7 Log CFU g-1. Metataxonomic analysis revealed that Lactococcus lactis and Leuconostoc mesenteroides occurred at high frequency in all the analyzed samples. Moreover, Lactiplantibacillus plantarum, Lactococcus piscium, and Lacticaseibacillus zeae were also detected. The most represented VOCs were carboxylic acids, carbonyl compounds, alcohols, and esters. In the analyzed cheese samples, significant relationships between bacteria and VOCs were also evidenced. Of note, research on the microbiological and chemical features, as well as on morpho-textural characteristics, of Queijo de Nisa PDO  cheese can increase the knowledge on the interactions between raw materials, environment, and autochthonous microorganisms, thus enabling protection of biodiversity and tradition.

Staphylococcus aureus artificially inoculated in mealworm larvae rearing chain for human consumption: Long-term investigation into survival and toxin production.

The present research reports the results of a long-term study (70 days) of the dynamics of Staphylococcus aureus artificially inoculated in a Tenebrio molitor rearing chain for human consumption. To this end, a rearing substrate consisting of organic wheat middlings was spiked with S. aureus to obtain three initial contamination levels, namely 1 (low level), 5 (medium level) and 7 (high level) Log colony forming unit per gram. Microbial viable counting coupled with metataxonomic analysis were performed to evaluate: i) the persistence and growth of S. aureus in the rearing substrate; ii) the colonization and growth of S. aureus in the insect larvae; iii) the occurrence and load of S. aureus in the frass (excrement from larvae mixed with substrate residues); iv) the presence of S. aureus enterotoxins in the rearing substrate, frass, and larvae. The results of the present study highlighted that wheat middlings contaminated with S. aureus do not represent a suitable environment for the multiplication of the pathogen, irrespective of the initial contamination level. Of note, frass originated from the larvae reared on contaminated wheat middlings might potentially represent a source of S. aureus, with cell loads depending on the initial contamination level. A complex resident microbiota was revealed by metataxonomic analysis. Interestingly, co-occurrence/co-exclusions analysis did not reveal associations between the target microorganism and the microbiota of wheat middlings, larvae, or frass. Considering safety aspects of larvae, the results overall collected suggested that, under the applied conditions, T. molitor represents an inhospitable or even hostile environment for S. aureus, with this latter showing counts below the detection limit in the larvae at the end of the 70-day rearing trial, irrespective of the initial contamination level. The results also suggested that a combination of bactericidal factors, including unfavorable environmental conditions (such as low aw of wheat middlings and frass), might have established in the rearing chain. Finally, the absence of staphylococcal toxins suggests that, even when S. aureus is present at high contamination levels, it is not able to produce toxins in wheat middlings, larvae, or frass.

Detecting Carbapenemases in Animal and Food Samples by Droplet Digital PCR.

BACKGROUND: The presence of carbapenemase-producing bacteria (CPB) in animal hosts and along the food chain may result in the development of reservoirs for human infections. Several CPB strains isolated from animals have been reported, suggesting that transmission and dissemination of the corresponding genes between humans and animals may occur. Animal and food samples have complex backgrounds that hinder the detection of CPB present in low concentrations by standard detection procedures. METHODS: We evaluated the possibility of detecting blaKPC, blaVIM, and blaOXA-48-like carbapenemases in 286 animal and food samples (faeces from farm and companion animals, raw meat, bivalve molluscs) by culture-based and standard molecular methods and by ddPCR. RESULTS: The proposed ddPCR managed to detect the target genes, also in samples resulting negative to standard methods. While the presence of blaKPC and blaVIM was detected in few samples (~3%), one third of the samples (n = 94/283) carried different variants of blaOXA-48-like genes. CONCLUSION: A specific and sensitive method such as ddPCR could be suitable to evaluate the current veterinarian and environmental situation and to assess the dynamic transmission and persistence of CPB between animals and humans and vice versa.

Liquid sourdough from stone-ground soft wheat (Triticum aestivum) flour: Development and exploitation in the breadmaking process.

The aim of the study was to set up a liquid sourdough obtained using stone-ground soft wheat (Triticum aestivum) flour to be exploited in breadmaking. Therefore, a Type II sourdough (dough yield = 350) was developed from a stable stone-ground wheat Type I sourdough (dough yield = 156) used as inoculum. Both sourdoughs were analyzed for lactic acid bacteria (LAB) viable counts, pH and total titratable acidity (TTA), LAB biodiversity by a combined culture-dependent and -independent approach (PCR-DGGE) and they were tested for their breadmaking ability. In addition, the chemical and rheological features and volatile organic compounds of the stone-ground soft wheat flour used in the experiment were investigated. The flour had a high protein content, good bakery properties and it also presented a rich aroma pattern characterized not only by the prevalence of green grass, flowery, and sweet aromas but also nutty, roasted and popcorn aromas. The sourdoughs I and II used in the trial were characterized by viable LAB counts, pH and TTA values typical of mature sourdoughs, i.e., approximately 9 log cfu gr-1 and mL, pH 3.9 and 10 mL 0.1 N NaOH. In addition, Levilactobacillus brevis and Companilactobacillus paralimentarius species represented the LAB stable microbiota of both sourdoughs. Both sourdoughs efficiently produce acceptable experimental breads characterized by different volatile profiles thus indicating that the type of sourdough fermentation significantly influenced the features of the final products. Overall, for the first time in the present study stone-ground wheat flour and bread have been characterized for their volatile aroma profile and sensory properties.

Fermentation of Microalgal Biomass for Innovative Food Production.

Fermentation is an ancient method used worldwide to process and preserve food while enhancing its nutraceutical profile. Alga-based fermented products have recently been developed and tested due to growing interest in healthy sustainable diets, which demands the development of innovative practices in food production, operating for both human health and Earth sustainability. Algae, particularly microalgae such as Arthrospira platensis, Chlorella vulgaris, and Dunaliella salina, are already cultivated as sources of food due to their valuable compounds, including proteins, pigments, lipids, carotenoids, polyunsaturated fatty acids, steroids, and vitamins. Due to their nutritional composition, functional diversity, and flexible metabolism, microalgae represent good fermentation substrates for lactic acid bacteria (LAB) and yeasts. This review presents an overview of the scientific studies on microalga fermentation underlining microalgae's properties and health benefits coupled with the advantages of LAB and yeast fermentation. The potential applications of and future perspectives on such functional foods are discussed.

Fate of Escherichia coli artificially inoculated in Tenebrio molitor L. larvae rearing chain for human consumption.

The edible insect food chain represents a relatively novel food-producing system; hence, associated biological risks still need to be exhaustively evaluated. In the present study, the dynamics of Escherichia coli during the whole living period of Tenebrio molitor larvae (from eggs to pupae) were studied. To this end, a rearing substrate consisting of organic wheat middlings was spiked with E. coli cells at two initial contamination levels: 1 log cfu g-1 (low) and 6 log cfu g-1 (high). Microbial viability counting coupled with metataxonomic analyses was used to assess i) the persistence and growth of E. coli in the rearing substrate (wheat middlings); ii) the colonization and growth of E. coli in the insect larvae; and iii) the occurrence and load of E. coli in the frass (excrement from larvae mixed with substrate residues). The results highlighted a very limited persistence of the pathogen in all analyzed samples. In more detail, the results suggested that when E. coli was present at very low levels in the eggs of the insect, the pathogen was not able to reach concerning levels in the larvae. Moreover, when E. coli was present in the wheat middlings used for rearing, the environmental conditions of the substrate (low aw values) were not favorable for its survival and multiplication, irrespective of the presence of the larvae and their frass. Surprisingly, under the conditions applied in the present study, the larvae fed wheat middlings contaminated with E. coli seemed to be inhospitable or even hostile environments for microbial survival or multiplication. To explain the low levels of E. coli cells in the larvae reared in the present study, many factors can be considered, including the immune response of the host, microbial composition and interactions established in the gut of larvae, and insect species. Of note, part of the major fraction of the microbiota of larvae at the end of rearing was represented by Lactococcus, thus suggesting a possible effect of this lactic acid bacterium on E. coli decay. Further research is needed to better clarify the interactions between E. coli and the insect gut, as well as the interactions established among the target microorganism and those naturally harbored by the insect gut.

Microbiological safety and stability of novel green sauces made with sea fennel (Crithmum maritimum L.).

Due to consumers' demand for ready-to-eat foods, in the last decades production of sauces has shifted from home-made to commercial practice. Besides to palatability and nutritional value, safety and stability are key issues of industrial sauces. The present study was aimed at evaluating the microbiological stability and safety of industrial-scale prototypes of two novel green sauces made with sea fennel (Crithmum maritimum L.) as the main ingredient. To this end, accelerated shelf-life and microbial challenge tests were performed to assess: (i) the microbiological shelf-stability of the two sauces stabilized by heat treatments commonly applied at industrial scale to inactivate vegetative cells of spoilage microorganisms and pathogens in vegetable preserves (F857=2min or F957=5min); and (ii) the inhibition of Staphylococcus aureus and Bacillus cereus in the sauce with pH = ∼ 4.6 and aw < 0.92, subjected to mild pasteurization (F757=1or2min). The results overall collected through the accelerated shelf-life tests clearly demonstrated the microbiological shelf-stability during one month of storage at thermal abuse conditions of both the novel sauces assayed; moreover, the microbial challenge tests revealed that both mild heat treatments assayed were able to inactivate S. aureus; in addition, an inhibition of the growth of B. cereus was seen during storage at 37 °C. Results from this study are expected to be useful both from a scientific and technological standpoint, enabling efficient risk-based development of novel acidified food products.

Microbial diversity, morpho-textural characterization, and volatilome profile of the Portuguese thistle-curdled cheese Queijo da Beira Baixa PDO.

The aim of the present study was to characterize the bacterial and fungal communities naturally occurring in Queijo da Beira Baixa PDO cheese samples produced in Castelo Branco district (Beira Baixa Region, Portugal) through viable counts and metataxonomic analyses. Physico-chemical and morpho-textural analyses were also carried out, together with the analysis of volatile organic compounds (VOCs). In the analyzed samples, pH values ranged between 4.72 ± 0.15 and 5.85 ± 0.02, with values of lactic acid content comprised between 0.64 ± 0.00 and 1.95 ± 0.16 g 100 g-1. Specific volume of cheese ranged from 1.09 ± 0.08 to 1.32 ± 0.02 g mL-1. Texture profile analysis showed hardness ranging between 38.3 ± 9.6 N and 68.55 ± 7.5 N. As for lactic acid bacteria, presumptive lactococci, thermophilic cocci, and lactobacilli counts up to 9 Log cfu g-1. Coagulase-negative cocci showed counts up to 7 Log cfu g-1. Enterococci counts were up to 6 Log cfu g-1. Finally, counts of eumycetes showed values up to 4 Log cfu g-1. The results of metataxonomic analysis of bacteria showed the dominance of Lactococcus lactis in all the samples. Moreover, other taxa were detected, including Lactiplantibacillus plantarum, Loigolactobacillus coryniformis, Lactococcus piscium, Streptococcus thermophilus, and Lacticaseibacillus zeae. Mycobiota was characterized by the presence of Candida sake, Ustilago, Cladosporium variabile, Starmerella, Debaryomyces hansenii, and Pichia kluyveri. In the analyzed Queijo da Beira Baixa PDO cheese samples, carboxylic acids represented the most detected VOCs, followed by esters, carbonyl compounds, and alcohols.