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.

Knot formation and spread along the shoot stem in 13 olive cultivars inoculated with an indigenous pathobiome of 7 species of Pseudomonas including Pseudomonas savastanoi.

Olive knot is a widely spread disease among olive (Olea europaea L.) trees. Pseudomonas savastanoi pv. savastanoi is recognized as the primary causative agent of the disease however, recent evidence indicated that consortia of bacteria (pathobiome), may favor its development. Several factors are involved in the host-plant relationship and affect the intensity of the symptoms. Among these the presence of wounds, or damages to the plants' tissues may affect the intensity and propagation of the disease. It remains unknown whether or not bacteria move from an infected wound to another not infected one via shoot tissues. The present investigation focused on the susceptibility to olive knot of several cultivars after inoculating artificial wounds with selected Pseudomonas species, while spreading the disease from these to wounds on the same stem, that had not been purposefully inoculated. The pathobiome for the inoculum was prepared with 7 species of Pseudomonas (including Pseudomonas savastanoi pv. savastanoi), isolated from knot samples collected from two different, heavily infected olive orchards. The inoculation was done after the manual execution of 10 horizontal wounds on the stem of potted plants of 13 olive cultivars grown in the greenhouse. Only the lowest 5 wounds were inoculated. The inoculated wounds showed a maximum percentage of knots after 187 days. All 13 cultivars showed knots yet, the cultivar with the most severe disease level to Pseudomonas savastanoi pv. savastanoi was 'Rosciola colli Esini'. The metataxonomic analysis performed on the olive knots removed after 225 days confirmed the dominance of the inoculated species Pseudomonas savastanoi in all the assayed cultivars. The not inoculated wounds did not show the knot disease likely because the bacterium's inability to transmigrate from the inoculated wounds to the non-inoculated ones.

Characterization of potentially health-promoting constituents in sea fennel (Crithmum maritimum) cultivated in the Conero Natural Park (Marche region, Central Italy).

CONTEXT: Sea fennel (Crithmum maritimum L. [Apiaceae]) is an aromatic herb rich in bioactive molecules, such as polyphenols, with potential positive effects on human health. OBJECTIVE: This study aimed at the characterization of sea fennel secondary metabolites, focusing on the phenolic fraction. MATERIALS AND METHODS: Samples of whole sprouts, sole leaves and sole stems were subjected to accelerated solvent extraction with methanol, and the resulting extracts were analyzed by high­performance thin­layer chromatography, high-performance liquid chromatography, and liquid chromatography coupled with diode array detection and high-resolution mass spectrometry (LC-DAD-HRMS). RESULTS: HPTLC and HPLC analyses of sea fennel extracts showed similar chromatographic profiles among the tested samples, and the prevalence of chlorogenic acid within the phenolic fraction was verified. Ten hydroxycinnamic acids, including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C, 11 flavonoid glycosides, e.g., rutin, hyperoside, isoquercitrin, two triterpene saponins and two hydroxylated fatty acids, were detected and annotated via liquid chromatography coupled with diode array detection and high-resolution mass spectrometry. DISCUSSION AND CONCLUSIONS: The use of accelerated solvent extraction and LC-DAD-HRMS for the characterization of sea fennel secondary metabolites allowed the annotation of seven compounds newly detected in sea fennel, including triterpene saponins and hydroxylated fatty acids.

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.

Exploratory Study for Probiotic Enrichment of a Sea Fennel (Crithmum maritimum L.) Preserve in Brine.

Considering the increasing consumer demand for vegan and vegetarian health foods, different vegetables have been already exploited to produce non-dairy probiotic foods. In addition to being rich in bioactive compounds, sea fennel (Crithmum maritimum L.), also known as rock samphire, represents a valuable candidate in the production of probiotic-enriched foods, and, to the authors' knowledge, it has not yet been explored as carrier for probiotics. Hence, the present study was aimed at evaluating the survival of a commercially available probiotic formulation, SYNBIO®, and Lactiplantibacillus plantarum IMC 509 in an artificially acidified, pasteurized sea fennel preserve in brine during a refrigerated storage of 44 days. Despite slight reductions in the microbial loads, at the end of the storage, both the probiotic formulations showed loads higher than 7.0 Log CFU g-1 of sea fennel or mL-1 of brine, above the recommended administration dose to exert beneficial health effects. Thus, acidified sea fennel sprouts in brine represent a potential vehicle for probiotics delivery to humans.

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.

Tetracycline Resistance Genes in the Traditional Swedish Sour Herring surströmming as Revealed Using qPCR.

Antibiotic resistance (AR) represents a global concern for human health. To the best of the authors' knowledge, no study addressing AR in surströmming, a traditional Swedish fermented herring, has been performed to date. The aim of the present research was to study the prevalence of tet(O), tet(S), tet(W), tet(K), and tet(M) genes encoding for resistance to tetracycline using quantitative PCR (qPCR) applied to ready-to-eat surströmming samples collected from three producers located in Sweden. The tet(M) gene was found in all the analyzed samples, and it was also the most abundant among the tested tet genes; moreover, tet(O) was the least frequently detected gene. As a general trend, all the analyzed samples showed a high occurrence of the target genes, with slight variations among the producers. A principal component analysis did not reveal any separation among the samples or producers. All the collected data allowed for a drawing of a first picture of the occurrence of tetracycline resistance genes in ready-to-eat surströmming samples. Since no differences among the samples manufactured by the different producers were observed, it is likely that the detected genes were homogeneously spread among the microbial species shared by the herrings used as raw materials. Moreover, it can be hypothesized that the presence of the detected genes was also the result of a selective pressure of the natural marine environment on the herrings' gut microbiota and, hence, on the pro-technological microorganisms responsible for the fermentation of surströmming. However, the contribution of the manufacturers to the contamination of the processed herrings cannot be excluded.

Antimicrobial Activity of Fermented Vegetable Byproduct Extracts for Food Applications.

To prevent foodborne diseases and extend shelf-life, antimicrobial agents may be used in food to inhibit the growth of undesired microorganisms. In addition to the prevention of foodborne diseases, another huge concern of our time is the recovery of agri-food byproducts. In compliance with these challenges, the aim of this work was to more deeply investigate the antimicrobial activity of extracts derived from fermented tomato, melon, and carrot byproducts, previously studied. All the fermented extracts had antimicrobial activity both in vitro and in foodstuff, showing even higher activity than commercial preservatives, tested for comparison against spoilage microorganisms and foodborne pathogens such as Salmonella spp., L. monocytogenes, and B. cereus. These promising results highlight an unstudied aspect for the production of innovative natural preservatives, exploitable to improve the safety and shelf-life of various categories of foodstuff.

The Microbial Diversity of Non-Korean Kimchi as Revealed by Viable Counting and Metataxonomic Sequencing.

Kimchi is recognized worldwide as the flagship food of Korea. To date, most of the currently available microbiological studies on kimchi deal with Korean manufactures. Moreover, there is a lack of knowledge on the occurrence of eumycetes in kimchi. Given these premises, the present study was aimed at investigating the bacterial and fungal dynamics occurring during the natural fermentation of an artisan non-Korean kimchi manufacture. Lactic acid bacteria were dominant, while Enterobacteriaceae, Pseudomonadaceae, and yeasts progressively decreased during fermentation. Erwinia spp., Pseudomonasveronii, Pseudomonasviridiflava, Rahnellaaquatilis, and Sphingomonas spp. were detected during the first 15 days of fermentation, whereas the last fermentation phase was dominated by Leuconostoc kimchi, together with Weissellasoli. For the mycobiota at the beginning of the fermentation process, Rhizoplaca and Pichia orientalis were the dominant Operational Taxonomic Units (OTUs) in batch 1, whereas in batch 2 Protomyces inundatus prevailed. In the last stage of fermentation, Saccharomyces cerevisiae, Candida sake,Penicillium, and Malassezia were the most abundant taxa in both analyzed batches. The knowledge gained in the present study represents a step forward in the description of the microbial dynamics of kimchi produced outside the region of origin using local ingredients. It will also serve as a starting point for further isolation of kimchi-adapted microorganisms to be assayed as potential starters for the manufacturing of novel vegetable preserves with high quality and functional traits.

Selection of cereal-sourced lactic acid bacteria as candidate starters for the baking industry.

The quality of sourdough bread mainly depends on metabolic activities of lactic acid bacteria (LAB). The exopolysaccharides (EPS) produced by LAB affect positively the technological and nutritional properties of the bread, while phytases improve the bioavailability of the minerals by reducing its phytate content. In the present study, a pool of 152 cereal-sourced LAB were screened for production of phytases and EPS for potential use as sourdough starter cultures for the baking industry. There was large heterogeneity in the phytase activity observed among the screened isolates, with 95% showing the ability to degrade sodium phytate on plates containing Sourdough Simulation Medium (SSM). The isolates Lactobacillus brevis LD65 and Lactobacillus plantarum PB241 showed the highest enzymatic activity, while the isolates ascribed to Weissella confusa were characterized by low or no phytase activity. Only 18% of the screened LAB produced EPS, which were distinguished as ropy or mucoid phenotypes on SSM supplemented with sucrose. Almost all the EPS producers carried one or more genes (epsD/E and/or epsA) involved in the production of heteropolysaccharides (HePS), whereas the isolates ascribed to Leuconostoc citreum and W. confusa carried genes involved in the production of both HePS and homopolysaccharides (HoPS). Monosaccharide composition analysis of the EPS produced by a selected subset of isolates revealed that all the HePS included glucose, mannose, and galactose, though at different ratios. Furthermore, a few isolates ascribed to L. citreum and W. confusa and carrying the gtf gene produced ß-glucans after fermentation in an ad hoc formulated barley flour medium. Based on the overall results collected, a subset of candidate sourdough starter cultures for the baking industry was selected, including Lb. brevis LD66 and L. citreum PB220, which showed high phytase activity and positive EPS production.

Microbiological characterization of Gioddu, an Italian fermented milk.

Gioddu, also known as "Miciuratu", "Mezzoraddu" or "Latte ischidu" (literally meaning acidulous milk), is the sole variety of traditional Italian fermented milk. The aim of the present study was to elucidate the microbiota and the mycobiota occurring in artisan Gioddu sampled from three Sardinian producers by combining the results of viable counting on selective culture media and high-throughput sequencing. Physico-chemical parameters were also measured. The overall low pH values (3.80-4.22) recorded in the analyzed Gioddu samples attested the strong acidifying activity carried out by lactic acid bacteria during fermentation. Viable counts revealed the presence of presumptive lactococci, presumptive lactobacilli and non-Saccharomyces yeasts. A complex (kefir-like) microbiota of bacteria and yeasts was unveiled through sequencing. In more detail, Lactobacillus delbrueckii was found to dominate in Gioddu together with Streptococcus thermophilus, thus suggesting the establishment of a yogurt-like protocooperation. Unexpectedly, in all the three analyzed batches from two out of the three producers Lactobacillus kefiri was also detected, thus representing an absolute novelty, which suggests the presence of bioactive compounds (e.g. exopolysaccharides) similar to those characterizing milk kefir beverage. Mycobiota population, studied for the very first time in Gioddu, revealed a more complex composition, with Kluyveromyces marxianus, Galactomyces candidum and Geotrichum galactomyces constituting the core species. Further research is needed to disclose the eventual occurence in Gioddu of probiotic cultures and bioactive compounds (e.g. exopolysaccharides, angiotensin-converting enzyme inhibitory peptides and antimicrobial compounds) with potential health-benefits for the consumers.

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.

Bacterial and Fungal Communities of Gioddu as Revealed by PCR-DGGE Analysis.

Gioddu is the sole variety of fermented milk originating in Italy. Despite the long history of consumption, Gioddu still represents an undisclosed source of microbial diversity. The present study was aimed to get an insight into the bacterial and fungal diversity of Gioddu samples collected from two artisan producers located in Sardinia. To this end 3 batches of Gioddu were collected from each producer and subjected to Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) analyses. Gioddu was produced with sheep milk in accordance with the local tradition. Regarding the bacterial population, a low biodiversity emerged. In more detail, the sole species Lactobacillus delbrueckii subsp. bulgaricus was detected in all the samples, irrespective of the producer or the batch. A more ample microbial diversity was highlighted for the fungal population that included closest relatives to Pichia cactophila, Kluyveromyces marxianus and Galactomyces candidum. Based on the results, the detected bacterial and fungal species generally clustered in accordance with the producer, irrespective of the batch considered. It is noteworthy that, Gioddu revealed several microbiological similarities with kefir beverage, thus suggesting, by analogy, potential health benefits related to its consumption. More research is needed to better clarify the microbiota composition of Gioddu by using more powerful metagenomic techniques.

Vegetable By-Product Lacto-Fermentation as a New Source of Antimicrobial Compounds.

BACKGROUND: One of the main objectives of the food industry is the shelf life extension of food products, taking into account the safety requirements and the preference of consumers attracted by a simple and clear label. Following this direction, many researchers look to find out antimicrobials from natural sources. METHODS: Tomato, carrot, and melon by-products were used as substrates for lactic acid fermentation using seven strains belonging to the Lactobacillus genus, L. plantarum, L. casei, L. paracasei, and L. rhamnosus. The obtained fermented by-products were then extracted and the antimicrobial activity toward fourteen pathogenic strains of Listeria monocytogenes, Salmonella spp., Escherichia coli, Staphylococcus Aureus, and Bacillus cereus was tested through agar well diffusion assay. RESULTS: All the extracts obtained after fermentation had highlighted antimicrobial activity against each pathogen tested. In particular, a more effective activity was observed against Salmonella spp., L. monocytogenes, S. aureus, and B. cereus, while a lower activity was observed against E. coli. CONCLUSION: Lactic acid fermentation of vegetable by-products can be a good strategy to obtain antimicrobials useful in food biopreservation.

Use of Dairy and Plant-Derived Lactobacilli as Starters for Cherry Juice Fermentation.

BACKGROUND: Lactic acid bacteria (LAB) exhibit a great biodiversity that can be exploited for different purposes, such as to enhance flavours or metabolize phenolic compounds. In the present study, the use of dairy and plant-derived LAB strains to perform cherry juice fermentation is reported. METHODS: The growth ability of Lactobacillus plantarum, Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus was studied in cherry juice. Profiling of sugars, organic acids and volatile compounds was performed by GC-MS (Gas Chromatography-Mass Spectrometry), while the phenolic fraction was characterized using UHPLC (Ultra High Performance Liquid Chromatography) equipped with a linear ion trap-mass spectrometer. RESULTS: Sucrose significantly decreased in all fermented samples as well as malic acid, converted to lactic acid by malolactic fermentation. The total amount of volatile compounds increased. Specifically, propyl acetate, an ester with fruit notes, reached the highest concentration in L. rhamnosus and L. paracasei (dairy strains) fermented juices. Phenolics were extensively metabolized: caffeic acid was converted into dihydrocaffeic acid, p-coumaric acid into 4-ethylphenol and phenyllactic acid was produced. CONCLUSION: Lactic acid fermentation confer fruit notes to the juice and enhance phenyllactic acids, especially employing dairy strains (L. rhamnosus and L. paracasei). The level of dihydrocaffeic acid, a compound with putative biological activity was also increased (in particular with L. plantarum).