Creating functional nanostructures: Encapsulation of caffeine into α-lactalbumin nanotubes

This work evaluated the stability and functionality of nanotubes obtained from α-lactalbumin (α-LA). α-LA nanotubes' structure was highly stable during a freeze-drying process but not after grinding. The ability of α-LA nanotubes to encapsulate caffeine, used as a model molecule, was evaluated. α-La nanotubes were highly effective for this purpose as encapsulation efficiency (%EE) was near 100% and loading capacity (%LC) near 10% at 1.5/20 and 2/20 ratios (caffeine/α-LA, w/w). α-LA nanotubes' structure was not affected by the presence of caffeine. Also, in general, refrigeration temperatures and neutral or alkaline conditions, under which the adverse effect of chelating agents was prevented, helped to stabilise α-LA nanotubes' structure and maintain caffeine encapsulated. At 8 °C and pH 7.5, in the presence of 75 μg mL− 1 of EDTA, > 50% of the caffeine remained encapsulated into α-LA nanotubes.Clara Fuciños gratefully acknowledge her Post-Doctoral grant (I2C 2014) from Consellería de Cultura, Educación e Ordenación Universitaria (Xunta de Galicia, Spain). Pablo Fuciños gratefully acknowledges his Marie Curie COFUND Postdoctoral Research Fellowship (Project No: 600375. NanoTRAINforGrowth - INL Fellowship programme in nanotechnologies for biomedical, environment and food applications). This study was supported by the Fundação para a Ciência e a Tecnologia (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), and the Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462)

Specific detection of viable Salmonella Enteritidis by phage amplification combined with qPCR (PAA-qPCR) in spiked chicken meat samples

Serovar Enteritidis represents 45.7% of all Salmonella reported human cases identified in Europe. Additionally, minced meat and meat preparations from poultry have a high level of non-compliance, regarding Salmonella regulation. In the current study, a novel method based on the amplification of the Salmonella bacteriophage vB_SenS_PVP-SE2, coupled with real-time PCR (qPCR), was developed and evaluated, for the rapid detection of viable Salmonella Enteritidis in chicken samples. The results obtained indicated that the qPCR method could detect down to 0.22 fg/L of pure virus DNA and a concentration of viral particles of 103pfu/mL. After a short bacterial recovery step, the addition of bacteriophages to spiked chicken samples indicated that 8cfu/25g could be detected within 10h, including the time for DNA extraction and qPCR analysis. Additionally, the evaluation of the performance parameters: relative sensitivity, specificity, accuracy, positive and negative predictive values, and index kappa of concordance, obtained values higher than 92%, and the acceptability limit values were within the limits. All these results demonstrate that the proposed methodology is a powerful tool for the rapid detection of viable Salmonella Enteritidis.This work was supported by the project Nanotechnology Based Functional Solutions (NORTE-01-0145-FEDER-000019), supported by NortePortugal Regional Operational Programme(NORTE2020), andby the “NanoBioSensor: Desenvolvimento de nanosensores para avaliação da qualidade microbiológica de produtos à base de fruta” (POCI-010247-FEDER-033925), supported bytheOperational ThematicProgram for Competitiveness and Internationalization (POCI), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF).info:eu-repo/semantics/publishedVersio

Rhamnolipids-based nanostructured lipid carriers: Effect of lipid phase on physicochemical properties and stability

In this work rhamnolipids were evaluated as surfactants for the production of nanostructured lipid carriers (NLCs). NLCs were produced by melt-emulsification using ultra-homogenisation followed by ultrasonication and different ratios of medium-chain-triglycerides and glycerol monostearate (lipid phase) were tested. NLCs presented sizes and polydispersity index values ranged between 97 and 120 nm and 0.200.26, respectively. Transmission electron microscopy observations confirmed the size and the spherical morphology of the NLCs. The thermal analysis and X-ray diffraction showed that the amount of solid lipid (glycerol monostearate) influences the melting, crystallisation and enthalpy of NLCs and their degree of crystallinity. Results showed that NLCs were more stable at 4 °C and the best formulation (1% of water phase, 0.05% of biosurfactant and solid:liquid ratio of 10:90) was stable for 30 days. This work showed the possibility of using rhamnolipids to produce NLCs and represent an important step for the development of lipid-based nanosystems using biosurfactants.Maria A. Azevedo (SFRH/BD/123364/2016) is the recipient of a fellowship from Fundação para a Ciência e Tecnologia (FCT, Portugal). This study was supported by FCT under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01- 0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145- FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. The authors would like to thank the H2020 MSCA-RISE project FODIAC—Food for Diabetes and Cognition (reference number 778388) and the Comissão de Coordenação e Desenvolvimento Regional do Norte (CCDR-N) project “Nanotechnology based functional solutions” (No. NORTE01- 0145-FEDER-000019). The authors would like to acknowledge to Oliver Schraidt from INL for his kind assistance using TEM.info:eu-repo/semantics/publishedVersio

Antiviral edible coatings and films: A strategy to ensure food safety

Background Pathological viral transmission via food has been a problem throughout humankind's evolutionary history, impacting food safety and public health. Fresh produce can be contaminated at any stage from pre- to post-harvest and frequently goes through little to no processing, imposing a high risk for contamination by foodborne viruses. Also, due to the globalization of the food industry and supply networks, the spread of enteric virus-related foodborne disorders has worsened. The current understanding of the transmission of viruses through contaminated foods needs more information regarding the potential infectivity, and it is essential to have effective ways to prevent viral transmission and minimize its adverse effects on human and animal health. Scope and approach This review addresses the global public health issue related to foodborne viruses and the current challenges of food safety. It provides an overview of food-grade and naturally occurring antiviral compounds with good antimicrobial activity and emphasizes how edible films and coatings with embedded antiviral agents can reduce the transmission of foodborne illness. Key findings and conclusions Antiviral edible films and coatings can be developed using plant-based compounds and their derived-products, like essential oils and extracts, with bioactive properties. Their use in food products and food contact materials can contribute to developing strategies to contain infectious outbreaks. Results show that these compounds interact with the viral particle, causing some damage to the virus integrity and affecting its infectivity. The development of antiviral edible films and coatings containing these bioactive compounds showed great potential against enteric viruses that cause foodborne illness, specifically norovirus and hepatitis A virus. However, some antiviral agents have also been shown to be very effective against other pathogenic viruses of great importance, some of which are not typically foodborne but can also be dangerous to humans.This work received funding from EIT Food (Grant agreement ID: 20400), the innovation community on Food of the European Institute of Innovation and Technology (EIT), a body of the EU, under Horizon 2020, the EU Framework Programme for Research and Innovation.info:eu-repo/semantics/publishedVersio

Alginate-based beads and core-shell capsules loaded with corn oil for potential delivery of functional compounds

info:eu-repo/semantics/publishedVersio

Ispitivanje fermentacijskih svojstava smeđe riže i rižinih mekinja radi uzgoja Lactobacillus plantarum NCIMB 8826

The growth on rice-based media of the probiotic strain Lactobacillus plantarum NCIMB 8826 isolated from the human gut has been investigated. Fermentation broths were obtained from the whole grain brown rice and rice bran of two Thai rice cultivars, RD6 (glutinous) and RD17 (nonglutinous). The rice used was not germinated and fermentations were carried out in a single step without growth supplementation. L. plantarum grew well in all tested broths, and a final biomass value of approx. 10.4 log CFU/mL was obtained. In addition, biomass production and substrate depletion were satisfactorily modelled using an unstructured mathematical model. There were no statistical differences observed among the four rice media. The results confirm that brown rice and rice bran are suitable substrates for the culture of the probiotic L. plantarum NCIMB 8826. Rice bran, currently a by-product of the traditional cereal processing industry, has shown similar fermentability to brown rice. This indicates that rice bran or rice bran extracts could be used in new probiotic food developments, while probably still maintaining other functional properties of the bran.U radu je ispitan rast soja mliječno-kiselih probiotičkih bakterija Lactobacillus plantarum NCIMB 8826, izoliranog iz crijeva čovjeka, u podlogama s rižom. Pripremljene su podloge od integralne (smeđe) riže i mekinja dvaju kultivara tajlandske riže, RD6 i RD17. Upotrijebljena je nenaklijala riža, a fermentacija je provedena u jednom koraku i bez dodatka stimulatora rasta. Soj L. plantarum rastao je dobro u svim podlogama, pa je postignut maksimalni prinos biomase od 10,4 log CFU/mL. Proizvodnja biomase i potrošnja supstrata uspješno su modelirane pomoću nestrukturiranoga matematičkog modela. Nije uočena statistički bitna razlika između rezultata dobivenih uzgojem soja u četiri različite podloge. Rezultati potvrđuju da su tajlandska integralna riža i rižine mekinje prikladne podloge za uzgoj probiotičkoga soja Lactobacillus plantarum NCIMB 8826. Utvrđeno je da rižine mekinje, nusproizvod dobivanja ljuštene (bijele) riže, imaju slična fermentacijska svojstva kao i smeđa riža. Stoga je zaključeno da se rižine mekinje ili ekstrakt rižinih mekinja mogu uspješno upotrijebiti za dobivanje novih probiotičkih proizvoda, a da pritom ne izgube ostala funkcionalna svojstva

Evaluation of antimicrobial effectiveness of pimaricin-loaded thermosensitive nanohydrogels in grape juice

Pimaricin-loaded poly(N-isopropylacrylamide) nanohydrogels with and without acrylic acid, were evaluated as food-spoilage inhibitors in a model system and a real food product: grape juice. Pimaricin was proposed as a non-allergenic alternative to sulphites for protecting juices against recontamination. However, pimaricin may degrade under conditions and treatments (heating, acidification, lighting) commonly applied in producing fresh juices. Nanohydrogel encapsulation may be a feasible procedure to avoid pimaricin degradation, improving its antimicrobial activity. Pimaricin-free nanohydrogels did not affect the growth of the indicator yeast either in the food model system or in grape juice. Conversely, pimaricin-loaded nanohydrogels effectively inhibited the growth of the indicator yeast. In some cases, the inhibition was extended even further than using free pimaricin. For instance, in the food model system, pimaricin-loaded nanohydrogels with acrylic acid (NPPNIPA-20AA(5)) prevented the yeast growth for more than 81 h while free pimaricin was only effective for 12 h. Despite pimaricin-loaded nanohydrogels without acrylic acid (NPPNIPA(5)) were able to reduce maximum yeast growth, as in all treatments with pimaricin, the extent of the inhibitory effect was not significantly (p>0.05) different to that achieved with free pimaricin. In grape juice, both free pimaricin and NPPNIPA-20AA(5) treatment completely inhibited the growth of the indicator yeast until the end of the bioassay. However, the latter provided similar inhibition levels using a smaller amount of pimaricin due to PNIPA-20AA(5) protection and its controlled release from the nanohydrogel. Therefore, nanohydrogel encapsulation may help to optimise antifungal treatments and decrease the incidence of food allergies.Funded by grant (MAT 2006-11662-CO3-CO2-C01/MAT 2010-21509-C03-01/EUI 2008-00115) from the “Ministerio de Educación y Ciencia” (Spain). Grant (SFRH/BPD/87910/2012) from the Fundação para a Ciência e Tecnologia (FCT, Portugal). Marie Curie COFUND Postdoctoral Research Fellow

3D printed functional cookies fortified with Arthrospira platensis: Evaluation of its antioxidant potential and physical-chemical characterization

In the last few decades, consumers' growing attention to the close relationship between health and nutrition is emerging as a new trend, mostly regarding the incorporation of natural ingredients into food. Among those ingredients, microalgae are considered as innovative and promising compounds, rich in valuable nutrients and bioactive molecules. In the present work, 3D printed cookies were fortified with the microalga Arthrospira platensis aiming at developing a new functional food with antioxidant properties. A. platensis antioxidants were recovered using ultrasound-assisted extraction in hydroalcoholic solutions. Ethanol/water and biomass/solvent ratios were optimised through a Design of Experiments (DOE) approach, using the antioxidant activity (ORAC and ABTS) and total phenolic content (TPC) as response variables. The highest ORAC, ABTS and TPC values were observed in the extract obtained with 0% ethanol and 2.0% biomass; thus, this extract was chosen to be incorporated into a printable cookie dough. Three different incorporation approaches were followed: (1) dried biomass, (2) freeze-dried antioxidant extract and (3) antioxidant extract encapsulated into alginate microbeads to enhance the stability to heat, light, and oxygen during baking and further storage. All dough formulations presented shape fidelity with the 3D model. The cookies had aw values low enough to be microbiologically stable, and the texture remained constant after 30 days of storage. Moreover, the extract encapsulation promoted an improvement in the ORAC value and colour stability when compared to all other formulations, revealing the potential of A. platensis for the development of a functional 3D food-ink.This work was funded by the European Union INTERREG Atlantic Area Programme and the European Regional Development Fund (ERDF) through the project “Enhance Microalgae: High added-value industrial opportunities for microalgae in the Atlantic Area” (Ref. EAPA_338/2016).info:eu-repo/semantics/publishedVersio

Polymeric stabilized oleogel-based emulsions aiming at food lipid profile tailoring

Decisive processing and technological requirements like saturated fat reduction and healthier fatty acids delivery are immediate objectives for food industry. In this work, our main focus was the development of gelled O/W emulsions, using oleogels as filling particles, that could act as healthier fat substitutes, namely for meat-based products.Fundação para a Ciência e TecnologiaCompete 2020FEDERinfo:eu-repo/semantics/publishedVersio

Faster monitoring of the invasive alien species (IAS) Dreissena polymorpha in river basins through isothermal amplification

Zebra mussel (Dreissena polymorpha) is considered as one of the 100 most harmful IAS in the world. Traditional detection methods have limitations, and PCR based environmental DNA detection has provided interesting results for early warning. However, in the last years, the development of isothermal amplification methods has received increasing attention. Among them, loop-mediated isothermal amplification (LAMP) has several advantages, including its higher tolerance to the presence of inhibitors and the possibility of naked-eye detection, which enables and simplifies its potential use in decentralized settings. In the current study, a real-time LAMP (qLAMP) method for the detection of Dreissena polymorpha was developed and tested with samples from the Guadalquivir River basin, together with two real-time PCR (qPCR) methods using different detection chemistries, targeting a specific region of the mitochondrial gene cytochrome C oxidase subunit I. All three developed approaches were evaluated regarding specificity, sensitivity and time required for detection. Regarding sensitivity, both qPCR approaches were more sensitive than qLAMP by one order of magnitude, however the qLAMP method proved to be as specific and much faster being performed in just 9 min versus 23 and 29 min for the qPCR methods based on hydrolysis probe and intercalating dye respectivelyThis work was supported by a partnership agreement project between the Confederación Hidrográfica del Guadalquivir and INL for the development of a system for early detection of zebra mussels through analysis of environmental DNA, and by project Nanotechnology Based Functional Solutions (NORTE-01-0145-FEDER-000019), supported by Norte Portugal Regional Operational Programme (NORTE2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund. S. A. acknowledges the Portuguese funding institution FCT – Fundação para Ciência e Tecnologia for Ph.D. scholarship SFRH/BD/140396/2018S