Unravelling microbial populations and volatile organic compounds of artisan fermented liver sausages manufactured in Central Italy.

The aim of the present study was to obtain information on the occurrence of bacteria and eumycetes in ready-to-eat fermented liver sausages manufactured by 20 artisan producers located in the Marche Region (Italy). To this end, culture-dependent analyses and metataxonomic sequencing were carried out. Physico-chemical parameters and volatilome of the fermented liver sausages were also studied. Finally, the presence of hepatitis E virus (HEV) was also assessed via real-time-RT-(q)PCR assays. Active microbial populations mainly represented by lactic acid bacteria, enterococci, coagulase-negative cocci, and eumycetes were detected. Enterobacteriaceae, Pseudomonadaceae, and sulfite-reducing anaerobes were not detected in most of the samples. Latilactobacillus sakei dominated in all the analyzed samples, reaching abundances up to 80%. Staphylococcus xylosus and Staphylococcus equorum were also detected. Among minority bacterial taxa, Weissella spp., Leuconostoc spp., Macrococcus caseolyticus, Brochothrix thermosphacta, Staphylococcus succinus, Lactobacillus coryniformis, Lactiplantibacillus plantarum, Lactococcus garviae, Psychrobacter spp., and Carnobacterium viridans were detected. The mycobiota was mainly composed by Debaryomyces hansenii that was present in all samples at the highest frequency. Among minority fungal taxa, Aspergillus spp., Penicillium spp., Kurtzmaniella zeylanoides, Candida spp., Yamadazyma spp., Scopulariopsis spp., Yarrowia spp., and Starmerella spp. were detected. Interestingly, associations between some taxa and some physico-chemical parameters were also discovered. The absence of HEV in all the samples attested a high level of safety. Finally, most of the VOCs detected in the analyzed fermented liver sausages belonged to six classes as: terpenoids, aldehydes, ketones, alcohols, esters, and acids. Nitrogen compounds, sulfur compounds, phenols, hydrocarbons, lactones, furans, and aromatic hydrocarbons were also identified. Several significant relationships were observed between mycobiota and VOCs.

Discovering microbiota and volatile compounds of surströmming, the traditional Swedish sour herring.

In this study, the microbiota of ready-to-eat surströmming from three Swedish producers were studied using a combined approach. The pH values of the samples ranged between 6.67 ± 0.01 and 6.98 ± 0.01, whereas their aw values were between 0.911 ± 0.001 and 0.940 ± 0.001. The acetic acid concentration was between 0.289 ± 0.009 g/100 g and 0.556 ± 0.036 g/100 g. Very low concentrations of lactic acid were measured. Viable counting revealed the presence of mesophilic aerobes, mesophilic lactobacilli and lactococci as well as halophilic lactobacilli and lactococci, coagulase-negative staphylococci, halophilic aerobes and anaerobes. Negligible counts for Enterobacteriaceae, Pseudomonadaceae and total eumycetes were observed, whereas no sulfite-reducing anaerobes were detected. Listeria monocytogenes and Salmonella spp. were absent in all samples. Multiplex real-time PCR revealed the absence of the bont/A, bont/B, bont/E, bont/F, and 4gyrB (CP) genes, which encode botulinic toxins, in all the samples analyzed. Metagenomic sequencing revealed the presence of a core microbiota dominated by Halanaerobium praevalens, Alkalibacterium gilvum, Carnobacterium spp., Tetragenococcus halophilus, Clostridiisalibacter spp. and Porphyromonadaceae. Psychrobacter celer, Ruminococcaceae, Marinilactibacillus psychrotolerans, Streptococcus infantis and Salinivibrio costicola were detected as minor OTUs. GC-MS analysis of volatile components revealed the massive presence of trimethylamine and sulphur compounds. Moreover, 1,2,4-trithiolane, phenols, ketones, aldehydes, alcohols, esters and long chain aliphatic hydrocarbons were also detected. The data obtained allowed pro-technological bacteria, which are well-adapted to saline environments, to be discovered for the first time. Further analyses are needed to better clarify the extent of the contribution of either the microbiota or autolytic enzymes of the fish flesh in the aroma definition.

Lesser mealworm (Alphitobius diaperinus) powder as a novel baking ingredient for manufacturing high-protein, mineral-dense snacks.

Increasing interest in consuming foods that are high in protein, vitamin, amino acid, and mineral contents is steering growth in the market for fortified snacks. The aim of the present study was to evaluate the use of lesser mealworm (Alphitobius diaperinus) powder (LP) (at 10 or 30% substitution for wheat flour) for the protein and mineral fortification of crunchy snacks (rusks). Hence, the technological, microbiological, nutritional, and sensory characteristics of the fortified rusks were evaluated. The protein content was enriched up to 99.3% in rusks with 30% substitution; moreover, a notable increase in the essential amino acids content was observed, with histidine fortification reaching up to 129.1% in rusks with 30% substitution. The incorporation of LP has led to an enrichment of almost all the minerals considered here, and especially Fe, P and Zn, with Zn showing fortification percentages of up to 300% in rusks with 30% substitution for LP. The experimental rusks showed pleasant sensory traits and low aw values. In view of the potential industrial manufacturing of insect-based rusks, the proposed product can be assigned to level 4 (validation in a laboratory environment) of the Technology Readiness Level (TRL) scale, and it is thus ready to be tested in a simulated production environment.

Is there any still undisclosed biodiversity in Ciauscolo salami? A new glance into the microbiota of an artisan production as revealed by high-throughput sequencing.

Ciauscolo is a fermented sausage with the Protected Geographical Indication (PGI) status. To disclose the microbial ecology of a model Ciauscolo salami manufacture during its natural fermentation, viable counting, amplicon-based sequencing and real-time PCR were applied. The volatilome during fermentation was also characterized. The results allowed previously undetected species to be discovered. The core microbiota was composed by Lactobacillus algidus, Leuconostoc carnosum, Lactobacillus sakei, Debaryomyces hansenii, Glomus hyderabadensis, Tilletiopsis washingtonensis, and Kurtzmaniella zeylanoides. Salmonella spp. and Listeria monocytogenes were absent in all the samples; moreover, multiplex real-time PCR revealed the absence of the target genes bont/A, bont/B, bont/E, bont/F, and 4gyrB (CP), encoding botulinic toxins. Volatilome, deeply depending on microbiological metabolism, was characterized by spices-derived components. Limonene, sabinene, α- and ß-pinene, 3-carene, and α-thujene were the most represented monoterpene hydrocarbons, whereas ß- and α-copaene were the most represented sesquiterpene hydrocarbons. Allyl methyl sulphide and diallyl disulphide were the major aliphatic sulphur compounds, together with diallyl sulphide and allyl methyl disulphide.

Microbial dynamics of model Fabriano-like fermented sausages as affected by starter cultures, nitrates and nitrites.

The present study promotes the valorization of Fabriano-like fermented sausages, which are central-Italy salami with an origin that dates to the early 17th century, for the possible future selection of autochthonous starter cultures to be used with respect to local traditions. To the best of the authors' knowledge, this study represents the first attempt to define the microbial dynamics in Fabriano-like fermented sausage and the effect of nitrates/nitrites and starter cultures on its natural bacterial biota. Culture and RNA-based techniques (RT-PCR-DGGE and Illumina sequencing) were used to assess the microbial ecology of model Fabriano-like fermented sausages together with the impact of starter cultures and different nitrate and nitrite concentrations. The meat batter was used to produce two batches of fermented sausages that were prepared as follows: i) without commercial starters or ii) with the use of starter cultures composed of Pediococcus pentosaceus and Staphylococcus xylosus. Each batch was further divided into three different batches with the addition of 0/0 mg kg-1 nitrate/nitrite, 75/60 mg kg-1 nitrate/nitrite and 150/125 mg kg-1 nitrate/nitrite to the first, second and third batch, respectively. The samples, which were produced in triplicate, were analyzed on the day of production and after 7, 21, and 42 days of ripening. Enterobacteriaceae counts were always higher in model Fabriano-like sausages produced without the use of starter cultures at all of the sampling times irrespective of the tested nitrate/nitrite concentrations. Lactobacilli counts were positively influenced by the starters, although this influence was not evident over time; moreover, the effect of nitrates and nitrites on the counts of lactobacilli differed over time. As a general trend, coagulase-negative cocci counts were apparently not influenced by the tested nitrate/nitrite concentrations. Regarding the effect of nitrates/nitrites on the microbial diversity revealed by RT-PCR-DGGE, the higher the concentration, the lower the presence of some genera/species such as Pseudomonas spp., Serratia liquefaciens and Staphylococcus spp. However, Illumina sequencing detected Pseudomonas spp. as a minority species after 7, 21 and 42 days of ripening irrespective of the nitrate/nitrite concentration. The presence of Staphylococcus species was highlighted by both RT-PCR-DGGE and Illumina sequencing at all of the stages of ripening, although its presence was massively detected in fermented sausages produced without the use of nitrates/nitrites at the end of ripening. Overall, the data collected clearly highlighted the dominance of Lactobacillus sakei in all of the fermented sausages during ripening (from day 7 to day 42) and irrespective of the nitrate/nitrite concentration and added starter cultures. Moreover, Pediococcus spp. was principally detected in model Fabriano-like fermented sausage with added starter cultures irrespective of the nitrate/nitrite concentration.

Experimental accelerated shelf life determination of a ready-to-eat processed food.

The most direct way to estimate the shelf life of a product is to conduct simulation tests which are time consuming and expensive. Conversely, accelerated shelf life tests can be successfully used for stable products having long expected shelf life. The aim of the study was directed to verify the possibility to apply an accelerated shelf life test to perishable food products having a short-expected shelf life, such as a new ready-to-eat processed food preparation, composed mainly by cereals, tuna and chicken, packed in thermo-sealed trays and pasteurised. Different samples of the product were stored in thermal abuse conditions, collected periodically and subjected to determinations of TVB-N, pH and sensorial characteristics. Q10 and activation energy were calculated allowing to obtain a predictive evaluation of the product shelf life at the 4°C recommended temperature. The product shelf life was assessed at 26 days vs the 30 days expected by the manufacturer, showing the possibility to apply successfully ASLT for products having short shelf life, saving both time and money.