Advances in chemical and biological methods to identify microorganisms—from past to present

Fast detection and identification of microorganisms is a challenging and significant feature from industry to medicine. Standard approaches are known to be very time-consuming and labor-intensive (e.g., culture media and biochemical tests). Conversely, screening techniques demand a quick and low-cost grouping of bacterial/fungal isolates and current analysis call for broad reports of microorganisms, involving the application of molecular techniques (e.g., 16S ribosomal RNA gene sequencing based on polymerase chain reaction). The goal of this review is to present the past and the present methods of detection and identification of microorganisms, and to discuss their advantages and their limitations.C.F.R. would like to thank the Portuguese Foundation for Science and Technology (FCT–Portugal) for the C.F.R. for the project UID/EQU/00511/2019—Laboratory for Process Engineering, Environment, Biotechnology, and Energy—LEPABE funded by national funds through FCT/MCTES (PIDDAC) and N.M. for the Strategic project ref. UID/BIM/04293/2013 and “NORTE2020 - Programa Operacional Regional do Norte” (NORTE-01-0145-FEDER-000012)

Alimentos funcionais com ingredientes inovadores com origem em algas e cogumelos

Doutoramento em QuímicaRecursos naturais, como algas e cogumelos podem ser usados para a obtenção de novos produtos, oferecendo uma forma alternativa e sustentável de originar novos alimentos ou ingredientes funcionais com propriedades biológicas que podem ajudar a desenvolver novas estratégias baseadas na saúde preventiva. A principal força motriz desta tese foi encontrar em algas e cogumelos, extratos com compostos bioativos naturais que possam ser usados no desenvolvimento de novos alimentos funcionais. Com este objectivo, foi efectuada uma abordagem integrada e vários passos foram seguidos numa ordem cronológica. Em primeiro lugar, selecionou-se um conjunto de espécies de algas marinhas assim como um conjunto de espécies de cogumelos comestíveis que foram caracterizados quanto à sua composição química e compostos bioativos potenciais. Tendo sido realizada a sua caracterização, surgiu a necessidade de se aplicar metodologias de extracção adequadas, rentáveis e amigas do ambiente, capazes de extrair ingredientes naturais biologicamente ativos de interesse tanto de algas como de cogumelos. Este objetivo nesta fase levou à segunda etapa do trabalho experimental que envolveu o estudo de várias técnicas de extração (extração com água quente, extração assistida por enzimas e por ultra-sons e extracção sob alta pressão hidrostática), a fim de se caracterizar plenamente o potencial das diferentes fontes naturais, introduzindo em simultâneo diferentes seletividades e eficiências, preservando o máximo de bioatividade. A etapa seguinte desta pesquisa integrada esteve relacionada com a caracterização química mais aprofundada de compostos bioativos presentes nos quatro extratos enzimáticos selecionados de algas (extrato S. muticum obtido com Alcalase e extrato O. pinnatifida obtido com Viscozyme) e de cogumelos (extratos de Ph. nameko obtidos com Celulase e com Flavourzyme) onde as propriedades biológicas alvo foram confirmadas ou verificadas como as mais promissoras. Os extratos selecionados com potenciais propriedades biológicas, após a caracterização química foram avaliados no que respeita à sua estabilidade in vitro para confirmar e consolidar o seu potencial biológico e ser mais explorado na perspectiva do alimento funcional. A última etapa deste trabalho envolveu o desenvolvimento de um novo alimento funcional incorporando-se os dois extratos mais promissores e validados previamente (extrato O. pinnatifida obtido com Viscozyme e extrato Ph. nameko obtido com Flavourzyme) num creme lácteo para barrar, procedendo-se à avaliação do seu potencial biológico e tecnológico. Um creme lácteo de barrar funcional, combinando requeijão e iogurte tipo grego com incorporação dos extratos selecionados foi formulado e explorado com sucesso. O desenvolvimento de alimentos funcionais, ou mesmo de nutracêuticos, a partir de extratos de algas e de cogumelos comestíveis é viável podendo-se estender o estudo da incorporação destes extratos em outros tipos de alimentos como bebidas ou sorvete.Natural resources such as seaweed and mushrooms can be used to obtain new products, providing an alternative and sustainable manner to provide new functional foods or ingredients with biological properties that may help to develop new strategies based on preventive health. The main driving force of this thesis was to find in seaweeds and mushrooms, extracts with natural bioactive compounds to be used in the development of new functional foods. In order to do so, an integrated approach was established and the various steps were followed chronologically. Firstly, a set of edible seaweeds species and a set of edible mushrooms species were characterised for their proximate composition and potential bioactive compounds. Once the characterization was achieved the need for suitable, fast, cost-effective and environmentally friendly extraction methodologies capable of extracting the biologically active natural ingredients of interest from both the seaweeds and mushrooms led to the second stage. This stage involved the study of several extraction techniques (hot water extraction; enzyme- and ultrasound-assisted extraction and high hydrostatic pressure) in order to fully characterize the potential of the different natural sources, introducing different extraction selectivity and efficiency while aiming at maximum preservation of bioactivity. The next stage in this integrated research was related with the deeper chemical characterization of the bioactive components present in the four enzymatic selected extracts from seaweeds (S. muticum extract obtained with Alcalase and O. pinnatifida extract obtained with Viscozyme) and mushrooms (Ph. nameko extracts obtained with Cellulase and with Flavourzyme) where target biological properties were confirmed or found to be more promising. The selected extracts with potential biological properties, following the chemical characterization went through the evaluation of in vitro stability to confirm and consolidate its biological potential to be further explored within the functional food perspective. The last stage of this thesis involved the development of a new functional food by incorporating the two most promising and validated extracts (O. pinnatifida obtained with Viscozyme and Ph. nameko obtained with Flavourzyme) in a spreadable dairy cream with assessment of their biological and technological potential. A functional dairy spreadable dairy cream combining whey cheese and greek type yoghurt with incorporation of the selected extracts was successfully formulated and explored. The development of functional foods, or even nutraceuticals, from edible seaweed and mushroom extracts is feasible and could be extended studying the incorporation of these extracts in other types of food such as beverages or ice cream

Électro-activation de solutions aqueuses de lactate et ascorbate de calcium et étude de leurs effets antibactériens sur les cellules végétatives et spores de Bacillus cereus ATCC 14579

Depuis la vulgarisation de certains concepts comme la globalisation ou la mondialisation, le secteur agroalimentaire a connu une expansion fulgurante et un engouement incessant pour la commercialisation d’aliments entre les peuples à travers le monde. Ce phénomène, contribuant significativement à l’accroissement économique des marchés, n’est toutefois pas sans risque. Pendant ce temps, les dangers de sources microbiologiques, notamment les pathogènes, sont véhiculés par des matrices alimentaires et voyagent d’un pays à l’autre, ce qui augmente le risque de contamination pour les consommateurs. Conséquemment, on assiste à une augmentation des cas d’allergies alimentaires, d’intoxications ou de toxi-infections alimentaires dont les agents étiologiques peuvent venir des quatre coins du monde. À cet effet, les organismes réglementaires comme l’Agence canadienne d’inspection des aliments (ACIA), Santé Canada, la Food and Drug Administration (USFDA) américaine ou d’autres autorités internationales compétentes comme l’Organisation des Nations unies pour l’alimentation et l’agriculture(FAO) et l’Organisation mondiale de la santé (OMS) multiplient leurs efforts afin de mettre en place des normes et politiques réglementaires pour aider l’industrie agroalimentaire à renforcer les contrôles depuis la fabrication jusqu’à la commercialisation des aliments. Les dangers microbiologiques venant de pathogènes comme Bacillus cereus demeurent un risque de santé publique majeur qu’il faut maîtriser afin d’assurer la protection des consommateurs. Bien que de nombreuses techniques de contrôle (e.g., additifs alimentaires, haute pression hydrostatique, rayonnements ionisants, procédés thermiques, etc.) ont été développées et utilisées pour assurer la salubrité et l’innocuité des aliments, dans certains cas cela n’a pas permis de produire des aliments totalement exempts de bactéries responsables de la dégradation/altération des aliments et de pathogènes causant des intoxications alimentaires comme c’est le cas avec B. cereus. En effet, cette bactérie pathogène est ubiquitaire, aérobie et anaérobie facultative. Elle est capable de produire dans une grande variété d’aliments et d’ingrédients comme les épices des spores très résistantes ainsi que différents types de toxines pouvant causer la diarrhée, la nausée, le vomissement et même la mort. Dans cette optique, et vue la grande difficulté à maitriser la contamination des aliments causée par ce pathogène, l’objectif général de cette recherche a été d’utiliser la technologie d’électro-activation, une branche appliquée de l’électrochimie qui s’intéresse notamment à la réactivité des solutions aqueuses, comme méthode alternative et potentiellement efficace pour lutter contre B. cereus afin de produire des aliments plus sécuritaires avec une grande valeur nutritionnelle et organoleptique. Pour y parvenir, des solutions aqueuses de sels d’acides organiques de lactate de calcium, d’ascorbate de calcium et de leur mélange équimolaire ont été électro-activées dans un réacteur soumis à un courant électrique continu avec des intensités de l’ordre de 250, 500 et 750 mA pendant un maximum de temps de 30 minutes afin de produire les acides organiques conjugués respectifs; de l’acide lactique et de l’acide ascorbique. Dans la première partie de ce travail de recherche, les caractéristiques physicochimiques (e.g., pH, acidité titrable, pKa) des solutions électro-activées (SÉA) ont été étudiées et leurs profils moléculaires comparés à ceux d’acides standards respectifs en utilisant différentes techniques (e.g., FTIR, HPLC, DSC, DPPH), ce qui a permis de confirmer la production d’acides organiques conjugués respectifs des sels utilisés. Ces SÉA avaient un pH très bas, une acidité titrable élevée, notamment pour l’ascorbate de calcium et le mélange. En plus, une activité antioxydante élevée a été observée pour la solution électro-activée d’ascorbate de calcium et du mélange. Dans la deuxième partie de l’étude, les SÉA traitées à 250, 500 et 750 mA pendant 10, 20 et 30 min ont été retenues pour être mises en contact avec des cellules végétatives de Bacillus cereus ATCC 14579 en conditions modèles (contact direct) afin d’évaluer leurs effets antimicrobiens sur ce pathogène. Les cellules ont été testées en contact direct avec les SÉA pendant 5, 30 et 60 secondes. Le même traitement a été également réalisé par contact direct avec des acides organiques standards (lactique, ascorbique) pendant 5, 30, 60, et 120 secondes afin de faire des comparaisons. Les SÉA et les acides organiques standards correspondants avaient les mêmes valeurs d’acidité titrable. Par la suite, les cellules ont été observées au microscope (coloration au bleu de méthylène et fluorescence) afin d’évaluer les effets inhibiteurs/destructeurs de ces solutions. Également, les SÉA ont été diluées avec de l’eau distillée pour obtenir des solutions possédant 10 à 90% de l’acidité titrable (force) initiale pour être ensuite testées contre les cellules de B. cereus. Les résultats ont démontré que toutes les SÉA avaient une grande efficacité contre les cellules végétatives de B. cereus. Également, même à des taux de dilution représentant en moyenne 20% de la force initiale des SÉA, l’effet antimicrobien était très élevé pour les différentes solutions. L’observation de B. cereus au microscope a permis de confirmer les effets létaux des SÉA. Dans ce volet avec des cellules végétatives de B. cereus, l’efficacité des SÉA a été estimée à une réduction de 4–7 log UFC/mL. En plus, il a été démontré que le pouvoir antibactérien des SÉA était nettement plus élevé que celui des acides lactiques et ascorbiques standards (conventionnels). Dans la troisième partie de cette étude, des solutions électro-activées de lactate de calcium, d’ascorbate de calcium et de leur mélange équimolaire à 750 mA pendant 30 minutes ont été retenues et utilisées contre des spores de Bacillus cereus ATCC 14579 en conditions modèles et dans du saumon Atlantique frais. Les spores traitées ont été analysées à l’aide de microscopes électroniques à balayage et à transmission pour évaluer les effets sporicides des SÉA. Les résultats obtenus ont clairement montré un grand pouvoir sporicide des SÉA utilisées sur les spores de B. cereus avec une réduction de 7 à 9 log en utilisant une population initiale de spores de 10⁹ UFC/mL, dépendamment des conditions évaluées; à savoir : en contact direct (2–30 min), dans du saumon utilisé comme matrice alimentaire(2–7 min), ainsi qu’en combinaison avec de la chaleur modérée de 60, 70, 80 et 90 °C pendant 0.5–2 min. Également, il a été observé que la capacité sporicide des SÉA augmentait avec la température et le temps de contact. La microscopie électronique à balayage et à transmission a permis de constater que les SÉA pouvaient provoquer la destruction totale des cellules de B. cereus, et notamment la perforation de la membrane (cortex et manteau), ainsi que le reflux de différentes composantes de la structure des spores de B. cereus. Tenant compte des résultats obtenus dans cette étude, nous pouvons conclure que les solutions électro-activées à base de lactate de calcium, ascorbate de calcium et leur mélange, notamment celles électro-activées à 750 mA–30 min, pourraient être d’une grande contribution afin de renforcer la capacité de l’industrie alimentaire à lutter contre B. cereus ATCC 14579 et de produire des aliments plus sécuritaires pour le consommateur.Since the popularization of concepts like globalization, the agri-food sector has experienced a huge expansion and a ceaseless craze for the marketing of food between the peoples worldwide. This phenomenon, contributing significantly to the economic growth of the markets, is not without risk, however. Meanwhile, microbiological hazards, including pathogens, are carried through food matrices and travel from one country to another, increasing the risk of contamination for consumers. Consequently, we are also witnessing an increase in cases of food allergies, foodborne illnesses and outbreaks, with etiological agents coming from all over the world. Thus, regulatory organisms such as Canadian Food Inspection Agency (CFIA), Health Canada, United States Food and Drug Administration (USFDA) or competent international authorities like Food and Agriculture Organization of the United Nations (FAO) and World Health Organization (WHO) are stepping up efforts to put in place regulatory standards and policies in order to help the food industry to strengthen controls from the processing to the marketing of foods. Microbiological hazards from pathogens like Bacillus cereus remain a major public health risk that must be controlled in order to ensure consumers protection. Although many techniques of control (e.g., food additives, high hydrostatic pressure, ionizing radiation, thermal processes, etc.) have been developed and used to ensure the safety and security of foods, in some instance this has not allowed to produce food products that are completely free of bacteria responsible for degradation/spoilage of food and pathogens causing food poisoning as is the case with B. cereus. Indeed, this pathogenic bacterium is ubiquitous, aerobic and facultative anaerobic. It is able to produce, in a wide variety of foods and ingredients such as spices, highly resistant spores as well as different types of toxins that can cause diarrhea, nausea, vomiting, and even death. In this context, and given the great difficulty in controlling the contamination of food caused by this pathogen, the general objective of this research was to use the electro-activation technology, an applied branch of electrochemistry which is particularly interested in the reactivity of aqueous solutions, as an alternative and potentially effective method to fight against B. cereus in order to produce safer foods with high nutritional and organoleptic values. To achieve this, aqueous solutions of organic acid salts of calcium lactate, calcium ascorbate and their equimolar mixture were electroactivated in a reactor subjected to a direct electric current with intensities of 250, 500 and 750 mA for a maximum time of 30 minutes in a bid to produce the respective conjugated organic acids, lactic acid and ascorbic acid. In the first part of this research work, the physicochemical characteristics (e.g.,pH, titratable acidity, pKa) of the electro-activated solutions (EAS) were studied and their molecular profiles compared to those of respective standard acids using different techniques (e.g., FTIR, HPLC, DSC, DPPH), which helped to confirm the production of conjugated organic acids from the respective salts used. These EAS had a very low pH, a high titratable acidity, particularly for the calcium ascorbate and the mixture. In addition, a high antioxidant activity was observed for the electro-activated calcium ascorbate solution and the mixture. In the second part of the study, the EAS treated at 250, 500 and 750 mA for 10,20 and 30 min were selected to be brought into contact with vegetative cells of Bacilluscereus ATCC 14579 under model conditions (direct contact) in order to evaluate their antimicrobial effects on this pathogen. The cells were tested in direct contact with the EAS for 5, 30 and 60 seconds. The same treatment was also carried out by direct contact with standard organic acids (lactic, ascorbic) for 5, 30, 60, and 120 seconds in order to make comparisons. The EAS and the corresponding standard organic acids had the same titratable acidity values. There after, the cells were observed under microscope (Methylene blue and fluorescence) to evaluate the inhibitory / destructive effects of these solutions. Also, the EAS were diluted with distilled water to obtain solutions with 10 to 90% of the initial titratable acidity (strength) to be tested against B. cereus cells. The results demonstrated that all the EAS made were highly effective against the vegetative cells of B.cereus. Also, even at dilution rates averaging 20% of the EAS initial strength, the antimicrobial effect was very high for the different solutions. In addition, the microscopic observation of B. cereus has confirmed the lethal effects of EAS. In this part with the vegetative B. cereus cells, the efficacy of the EAS was estimated to a reduction of 4–7 log CFU/mL. In addition, the antibacterial power of the EAS has been shown to be significantly higher than that of the standard (conventional) lactic and ascorbic acids. In the third part of the study, electro-activated solutions of calcium lactate, calcium ascorbate and their equimolar mixture at 750 mA for 30 min were selected and used against the spores of Bacillus cereus ATCC 14579 under model conditions and in fresh Atlantic salmon. The treated spores were analyzed using scanning and transmission electron microscopes to evaluate the sporicidal effects of EAS. The results obtained clearly showed a great sporicidal power of the EAS used on B. cereus spores with a reduction of 7 to 9 log using an initial spore population of 10⁹ CFU/mL, depending on the conditions assessed; namely: in direct contact (2–30 min), in salmon used as a food matrix (2–7 min), as well as in combination with moderate heat of 60, 70, 80 and 90 ℃ for 0.5–2 min. Also, it was observed that the sporicidal capacity of the EAS increased with temperature and contact time. Scanning and transmission electron microscopy showed that the EAS could cause the total destruction of B. cereus cells, including perforation of the membranes (cortex and coat), as well as the reflux of different components of the structure of B. cereus spores. Taking into account the results obtained in this study, we can conclude that the electro-activated solutions made with calcium lactate, calcium ascorbate and their mixture, especially those electro-activated at 750 mA–30 min, could be of a great contribution to reinforce the capacity of the food industry to control B. cereus ATCC 14579 and produces safer foods for the consumer

Physicochemical, Sensory and Nutritional Properties of Foods Affected by Processing and Storage

Processing and storage can cause changes and interactions in food components that have effects on nutritional value, organoleptic characteristics or even food safety. This book includes 19 research works showing important and interesting advances, as well as new approaches, in this research topic. Four articles are dedicated to studying the effect of canning conditions (filling media and some ingredients) on the diverse parameters of quality for fish and pet foods. Three articles are devoted to studying the effects of dehydration (pre-treatments and drying procedures). One article is dedicated to monitoring the elaboration of a fermented and dehydrated product (sausage) using a portable NIRS device. The ninth article of this book studies the effect of low-dose electron beam irradiation on cooking quality, moisture migration, and thermodynamics, as well as the digestion properties of the isolated starches in newly harvested and dried rice. The next contribution studies the use of different preservatives to avoid the formation of undesirable volatile organic compounds in stracciatella cheese. Another article examines the impact of source material, kibble size, temperature, and duration on the efficiency of the aqueous extraction of sugars and phenolics from carob kibbles by conventional heat-assisted (HAE) and ultrasound-assisted (UAE) methods. In two articles, marinating with different extracts, alone or combined with other seasoning/conditioning methods, was essayed to tenderize beef or to improve the sensory quality of chicken leg and breast meat. The effect of various cooking methods on the quality, structure, pasting, water distribution and protein oxidation of fish and meat-based snacks is studied in the fourteenth article. The last five articles are dedicated to the study of the effects of storage on several foods (olive oil, blueberry, beetroot and Atlantic mackerel)

On the Mechanism of Electron Beam Radiation-Induced Modification of Poly(lactic Acid) for Applications in Biodegradable Food Packaging

Gemstone Team PRODUCEPoly(lactic acid) (PLA) is a biodegradable polymer used for food packaging. The effects of electron beam radiation on the chemical and physical properties of amorphous PLA were studied. In this study, amorphous, racemic PLA was irradiated at doses of 5, 10, 15, and 20 kGy in the absence of oxygen. Utilizing electron paramagnetic resonance spectrometry, it was found that alkoxyl radicals are initially formed as a result of C-O-C bond scissions on the backbone of the PLA. The dominant radiation mechanism was determined to be H-abstraction by alkoxyl radicals to form C-centered radicals. The C-centered radicals undergo a subsequent peroxidation reaction with oxygen. The gel permeation chromatography (GPC) results indicate reduction in polymer molecular mass. The differential scanning calorimetry and X-ray diffraction results showed a subtle increase in crystallinity of the irradiated PLA. Water vapor transmission rates were unaffected by irradiation. Further mechanical testing showed mechanical properties in line with reduced molecular mass. In conclusion, these results support that irradiated PLA is a suitable material for applications in irradiation of food packaging, including food sterilization and biodegradation

Novel Strategies for the Development of Healthier Meat and Meat Products and Determination of Their Quality Characteristics

Meat and meat products are in demand worldwide. However, new trends and innovations, driven by consumer interests and demands, have contributed to the development of healthier meat products, which are growing exponentially. Health is one of the main criteria for consumers when choosing the type of food they eat. This, along with more sustainable global production, changes in lifestyles, and nutritional ideologies among the population, has led to a growing demand in the market for healthier and more functional foods. This Special Issue has collected nine original research articles and one editorial. This book collects all-new, relevant information on the different strategies for modifying meat and meat products to make them healthier and more sustainable and the different quality control methods for their control. With this information, we hope to shed more light on this important topic. This SI will prove very useful and interesting to all those involved in the meat and meat product industry (livestock producers, processing companies, researchers, scientists, consumers, administration, etc.). Lastly, many of the studies featured in this SI leave the door open to future research in this area of growing interest, not only for the meat and meat product industry but for other foods as well

The physico-chemical properties of the Namibian pelagic fishes and seaweed polysaccharides.

Fish provides a major source of proteins and polyunsaturated fatty acids which are known to prevent coronary heart diseases. However, inherent characteristics such as colour and texture prevent the effective utilisation of some fish, especially fatty pelagics for human consumption. Instead, they are used for fish oil and fish meal production. For this reason and for enhancing the utilisation of valuable resources, the physico-chemical properties, which may be species specific, were investigated in relation to two Namibian species, namely, horse mackerel (Trachurus capensis) and snoek (Thyrsites atun). The results showed that horse mackerel (Trachurus capensis) and snoek (Thyrsites atun) contain high contents of palmitic acids (29%), oleic acids (25%), and the omega-3-polyunsaturated fatty acids, such as docosahexaenoic acids (DHA) (15%) and eicosapentaenoic acids (EPA) (9%). The studies also showed that optimum protein extractability was obtained with 5% NaCl at pH 7.2 and the highest gel strength at pH 6.5-7.0. CaCO3 had detrimental effect on the water-holding capacity when used above 50mg.l-1 in washing water. Agar and sodium alginate extracted from Namibian Gracilaria verrucosa (red seaweed) and Laminaria schinzii (brown seaweed) respectively, were also characterised for their physico-chemical properties. Both the nontreated and treated agars were observed by colorimetric method to contain low levels of sulphate, an observation also confirmed by NMR and Raman FTIR spectroscopy. Sodium alginate was found to contain high content of guluronic acid (75%) by NMR, a good index of strong gelling ability. The good functionalities obtained as a result of optimising the physicochemical properties of fish proteins and seaweed polysaccharides enabled an investigation of the interactions of the two components. Studies on the interactions of the both horse mackerel and snoek with either sodium alginate or agar were undertaken by rheology, differential scanning calorimetry and phase contrast microscopy. Large deformation and small deformation rheological studies showed a decrease in the gel strength of fish proteins with the addition of either polysaccharide at all combinations. The DSC results indicated a lowering of the denaturation temperature with the addition ratios of increasing amounts of sodium alginate to fish proteins. Although the mixture of fish proteins and agar resulted in the lowering of the denaturation temperature, the shift; was not as pronounced as that obtained with sodium alginate. The reason for the lowering of gel strength and G' values was attributed to phase separation of fish proteins and polysaccharides which was reflected by phase contrast microscopy studies. This investigation has resulted in the characterisation of valuable fish and seaweed polysaccharide resources from Namibia which could be used in combination for the development of new food products

Comparative analysis of biofortification effects on phenolic and glucosinolate profiles in Diploptaxis tenuifolia (L.) DC T&T hybrid

Wild rocket, scientifically known as Diplotaxis tenuifolia (L.) DC is a leafy green plant from the Brassicaceae family. Native to the Medditerian region, today this perennial plant is cultivated worldwide, owing to the distinctive pungent taste of its deeply lobed green leaves. Beyond their culinary uses, the leaves of wild rocket exhibit potential for pharmaceutical applications due to their rich bioactive compounds profile. These include vitamins, pigments (chlorophyll a and b, carotenoids), phenolics, and glucosinolates (GLSs), all exhibiting strong antioxidant and anti-cancer activities. This study aimed to evaluate the effects of biofortification, employing biostimulant Kelpak and two distinct foliar fertilizers (iron and potassium-enriched) in comparison to a control group, on the phenolic and glucosinolate content in the leaves of wild rocket hybrid T&T F1. Using an ultra-highperformance liquid chromatography (UHPLC) system, coupled with a quadrupole time-offlight mass spectrometry (Q-ToF-MS), a total of twenty-three plant phenolics was quantified, and the relative content of four glucosinolates was evaluated in 70% methanol extracts. Concerning the overall content of phenolic compounds, treatment involving iron application showed the highest levels at 3549.1 mg/kg of fresh weight (FW), while the control group exhibited the lowest content (2171.4 mg/kg FW). Moreover, the most prominent phenolics included quercetin-3,4'-di-O-hexoside-3'-O-(6"-sinapoyl)-hexoside (ranging from 527.2 to 651.8 mg/kg FW among groups) and 1,8-dipropoxyanthraquinone (278.3-558.5 mg/kg FW). The control and iron-enriched samples exhibited the presence of kaempferol-3,7,4'-tri-Ohexoside + HCOOH and isorhamnetin-3-O-hexoside-4'-O-gentobioside, in addition to the non-identified sinapic acid hydroxide in the Kelpak treated group. Additionally, the determination of relative GLSs content of glucosativin, glucoerucin, neoglucobrassicin, and DMB-GLS revealed that glucosativin was the dominant compound, with proportions ranging from 79.8% (Kelpak treatments) to 89.1% (iron treatments). In conclusion, selected hybrid biofortification treatments, notably through the use of iron foliar fertilizers, enhanced phenolic and glucosinolate leaf content, highlighting its potential significance in nutrition and medicine