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Optimization of the extraction conditions provides olive pomace extracts with improved bioactivities.

This study aims to study the kinetics and mechanisms of human adenovirus inactivation by electron beam. Human adenovirus type 5 (HAdV-5) was inoculated in two types of aqueous substrates (phosphate-buffered saline - PBS, domestic wastewater - WW) treated by electron beam at a dose range between 3 and 21 kGy. Samples were evaluated for virus infectivity, PCR amplification of fragments of HAdV-5 genome and abundance and antigenicity of the virion structural proteins. The maximum reduction in viral titre, in plaque-forming units (PFU) per millilitre, was about 7 and 5 log PFU/mL for e-beam irradiation at 20 kGy in PBS and 19 kGy in wastewater, respectively. Among the virion structural proteins detected, the hexon protein showed the higher radioresistance. Long (10.1 kbp) genomic DNA fragments were differently PCR amplified, denoting a substrate effect on HAdV-5 genome degradation by e-beam. The differences observed between the two substrates can be explained by the protective effect that the organic matter present in the substrate may have on viral irradiation. According to the obtained results, the decrease in viral viability/infectivity may be due to DNA damage and to protein alterations. In summary, electron beam irradiation at a dose of 13 kGy is capable of reducing HAdV-5 viral titres by more than 99.99% (4 log PFU/mL) in both substrates assayed, indicating that this type of technology is effective for viral wastewater disinfection and may be used as a tertiary treatment in water treatment plants. KEY POINTS: • The substrate in which the virus is suspended has an impact on its sensitivity to e-beam treatment. • E-beam irradiation at 13 kGy is capable of reducing by 4 Log PFU/mL the HAdV-5 viral titre. • The decrease in viral viability/infectivity may be due to DNA damage and to protein alterations.

This work aimed to establish an integrated approach to investigate the total phenolic content and antioxidant activities of dried skim camel and cow milk and their fractions. The milk fractions were obtained by acid or enzymatic coagulation followed by spray drying (inlet temperature/outlet temperature: 125 ± 2 °C/90 ± 2 °C) or freeze drying (−50 °C, 0.05 mbar) coupled or not to gamma irradiation (at 5, 11, 22 kGy). The results showed that the total phenolic content (measured in gallic acid equivalent, GAE) varied depending on the drying technique. The freeze-drying process corresponded to the highest values of total phenolic compounds, with 247.23 ± 2.08 mg GAE/100 g powder for the β-casein fraction of camel milk (βC CaM) and 621.13 ± 4.16 mg GAE/100 g powder for the β-casein fraction of cow milk (βC CoM). Compared to spray-dried fractions, freeze-dried fractions showed generally higher ferric reducing antioxidant power for both camel milk and cow milk. The highest values of free radical scavenging activity were seen in the spray-dried β-casein fractions of camel milk (βC CaM) and cow milk (βC CoM) and in the freeze-dried acid whey of cow and camel milk (AW CaM and AW CaM). Freeze-dried acid whey (AW CaM and AW CoM) appeared to be less sensitive to gamma irradiation at 5 and 11 kGy.

Bioactive potential of hazelnut husks was determined as a function of their cultivar source and extraction solvent. Hazelnut husks from four hazelnut cultivars (Butler, Grada de Viseu, Lansing and Morell) were picked in a hazelnut orchard at harvest and extracted with five solvents with different polarity: water, methanol, acetone, ethyl acetate and hexane. Phenolics were identified by HPLC–DAD and antioxidant activity was determined by three complementary methods: DPPH, FRAP and inhibition of lipid peroxidation. A total of 11 phenolics were identified in studied cultivars and grouped in five main classes namely, ellagitannin (ellagic acid), benzoic acids (gallic acid, protocatechuic acid and vanillic acid), flavonols (kaempferol-3,7-O-diglucoside, kaempferol-3-O-[6-acetylglucoside]-7-O-glucoside, kaempferol-3-O-[6acetylglucoside]-7-O-rhamnoside and quercetin-3-O-rutinoside), flavone (luteolin-7-O-rutinoside) and flavan-3-ol (epicatechin). Cultivar and extraction solvent influenced significantly (p

Due to the growing demand in society for healthier foods, scientific communities are searching and developing new ingredients. In this context, agro-industrial residues, which can have a negative impact on the environment, represent a natural source for bioactive compounds and their recovery can contribute to economic and environmental sustainability. Ionizing radiation is a clean and eco-friendly technology that can be used to improve the extraction of bioactive compounds. The aim of this review, after presenting general aspects about bioactive compounds in agro-industrial residues and radiation technologies, is to focus on the effects of ionizing radiation on the extraction of bioactive compounds from these residues and related bioactive properties. Irradiated residues were demonstrated to have enhanced bioactive characteristics that turn the prepared extracts suitable for applications in food industry, resulting in high-added-value products as well as reducing adverse impacts on the environment. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support to C2TN (UIDB/04349/2020), CIMO (UIDB/00690/2020), and J.M. (SFRH/BD/136506/2018); L.B. thanks FCT for the national funding, P.I., through the institutional scientific employment program-contract; FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_ 6_E. info:eu-repo/semantics/publishedVersion

Background Various strategies are needed to mitigate the negative impact on or to increase fruit quality. The effect of spraying kaolin (K), Ascophyllum nodosum (An) and salicylic acid (SA), in trees with and without irrigation, on quality and sensorial attributes of hazelnut (Grada de Viseu cultivar) was investigated during two consecutive years (2016 and 2017) in a commercial orchard located in Moimenta da Beira, Portugal. Results The treatments affected positively the biometric parameters nut and kernel weight, length, width, thickness and volume as well as the vitamin E level, antioxidant activity and content of some individual phenolics, such as protocatechuic acid, gallocatechin, catechin and epicatechin. The levels of amino acids in hazelnut kernels decreased in all the assayed treatments, while the kernel colour and sensorial attributes were not affected by the treatments. Hazelnut physical properties (nut and kernels), chemical and phytochemical composition and antioxidant activities were positively related. Conclusions The application of K, An and SA improved the hazelnut tree response to climate change, without compromising the hazelnut chemical and sensorial quality. Furthermore, due to the similar observations for the same treatments with and without irrigation, it can be stated that K, An and SA can be efficient and cost-effective tools to mitigate summer stress in rain-fed orchards. © 2020 Society of Chemical Industry.

Chestnut fruits, from Castanea sativa Miller species, have been previously characterized mainly to study the nutritional and biochemical parameters, with mineral contents receiving less attention from the researchers, although these minerals are essential for several mechanisms in human body. In this study, a detailed elemental compositional analysis was performed using Proton Induced X-ray Emission (PIXE) to characterize two varieties of Castanea sativa Miller chestnuts (Longal and Judia). This study comprises the composition of the outer shells (pericarp), inner shells (episperm) and the edible fruits. From our knowledge, this is the first time that the presence of up to 20 elements on the shells and the fruits are reported. Furthermore, by PIXE it was possible to quantify them, allowing significant improvements in the assessment of chestnuts composition. Essential and nonessential elements were quantified on a food item that is used mainly for human consumption but it is also incorporated in animal feeding. Authors acknowledge R. Pinheiro for sample preparation. This work was developed within the Coordinated Research Project D61024 ‘‘Development of New Applications of Machine Generated Food Irradiation Technologies’’ financed by the International Atomic Energy Agency (IAEA). IST authors gratefully acknowledge the FCT (Fundação para a Ciência e a Tecnologia) support through the UID/Multi/04349/ 2013 and UID/FIS/50010/2013 projects. info:eu-repo/semantics/publishedVersion

The aim of this work was to study the impact of ionizing radiation (gamma radiation and electron beam, 1–3 kGy) on the antioxidants of the strawberry, through evaluation of the total phenolic and L-ascorbic acid contents, antioxidant activity, and cytotoxicity (293 T and A549 cell lines) after storage at 4 °C for 15 days. Results showed that both gamma and electron beam radiation could preserve the phenolic content of strawberries during the storage. Both irradiation treatments induced the degradation on L-ascorbic acid content and indicated to increase the antioxidant activity mainly on strawberries treated at 2 kGy. Regarding cytotoxicity assay, non-tumor 293 T cells were more sensitive to strawberry extracts than A549 cells and ionizing radiation had no impact on the viability of both cell lines. These results pointed out that gamma and electron-beam radiation at 2 kGy can be used as safe and environmental friendly post-harvest treatments of strawberries to preserve their bioactive properties.

The environmental stability of enteric viruses and resistance to conventional treatments and common disinfectants, leads to their persistence in waters and food, causing serious implications on public health. Among non-thermal treatment methods, ionizing radiation is recognized as a useful and effective mean of disinfection. The objective of this study was to estimate the inactivation of enteric virus by gamma radiation in raw berry fruits, in order to evaluate the potential of this technology to be applied as a disinfection treatment. Fresh strawberries and raspberries were inoculated either individually with murine norovirus type 1 (MuNoV; as a human norovirus surrogate) and human adenovirus type 5 (HAdV) or with a viral pool of both viruses, and irradiated in a Co-60 equipment at doses of 1 kGy up to 11 kGy. The infectivity of viral particles of MuNoV and HAdV was assessed by plaque assay using Raw 264.7 and A549 cells, respectively. A 2 log PFU/g reduction on MuNoV and HAdV titers was obtained after treatment with a dose of 4 kGy for both fruits. However, non-linear inactivation survival curves were obtained for MuNoV and HAdV in fresh fruits, leading to the detection of infective viral particles at a dose of 11 kGy. The irradiation process indicated virucidal potential, although the estimated gamma radiation dose to attain food safety (> 7 kGy) would compromise the preservation of food quality. Nevertheless, the irradiation technology could be an effective virus mitigation tool to treat polluted waters, which are a major vehicle of contamination for fresh produce.

This study aims to assess the airborne bioburden of rural and urban Portuguese Primary Health Care Centers (PHCC) using active and passive sampling methods and identify the potential differences in airborne microbiota between both environments. The highest total aerobic mesophilic bacterial load in indoor air were found in the Vaccination Room (448 CFU.m−3) in the Rural PHCC and in the Waiting Room (420 CFU.m−3) for Urban PHCC. The total coliforms contamination level in indoor air was detected only in the Cleaning Supplies Room (4 CFU.m−3) in the Urban PHCC. The most frequent bacteria genera identified was Micrococcus (21% Rural PHCC; 31% Urban PHCC). The surface samples showed a highest total aerobic mesophilic bacterial contamination in the Treatment Room (86 × 103 CFU.m−2) from the Rural PHCC and in the Front Office (200 × 103 CFU.m−2) from the Urban PHCC. The electrostatic dust cloth (EDC) samples showed a highest bacterial load in the Urban PHCC. Total aerobic mesophilic bacterial load in settled dust and in the Heating, Ventilating and Air Conditioning (HVAC) filter samples in the Urban PHCC (8 CFU.g−1 and 6 × 103 CFU.m−2) presented higher values compared with the Rural PHCC (1 CFU.g−1 and 2.5 × 103 CFU.m−2). Urban PHCC presented higher bacterial airborne contamination compared with the Rural PHCC for the ma-jority of sampling sites, and when compared with the indoor air quality (IAQ) Portuguese legislation it was the Rural PHCC in two sampling places who did not comply with the established criteria. publishersversion published

Portobello variety of Agaricus bisporus mushrooms, appreciated for its taste, makes it desirable to be eaten fresh and also as flour in soups and gravies. Gamma and electron-beam radiation at four doses (1, 2, 5, and 10 kGy) were used to analyze its preservation effect on Portobello mushroom flour. A proximate analysis, as well as the impact on fatty acids, tocopherols, soluble sugars, organic acids, and ergosterol profiles, were performed every 3 months, during a storage period of 1 year. Gamma rays preserved mannitol (most abundant soluble sugar) over the 12 months, while electron beam radiation preserved organic acids. No significant changes were sought for any radiation type, and the slight changes extracted from the estimated marginal means reveal a tendency for irradiation as having preserving effects of nutrients and other important molecules. Thus, both irradiation types, up to 10 kGy are suitable for preservation of A. bisporus Portobello flour. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/AGR/00690/2020), C2TN (UID/Multi/04349/2013) for R.V.C. Cardoso’s grant (SFRH/BD/137436/2018), M. Carocho’s research contract. L. Barros and A. Fernandes also thank FCT’s funding through the institutional scientific employment program-contracts. This work is funded by the European Structural and Investment Funds (FEEI) through the Regional Operational Program North 2020, within the scope of Project Mobilizador ValorNatural R . Acknowledgments are also due European Agricultural Fund for Rural Development (EAFRD), through the Rural Development Program (PDR2020), within the scope of Project MicoCoating (PDR2020-101-031472). info:eu-repo/semantics/publishedVersion

Biological therapies, such as recombinant proteins, are nowadays amongst the most promising approaches towards precision medicine. One of the most innovative methodologies currently available aimed at improving the production yield of recombinant proteins with minimization of costs relies on the combination of in silico studies to predict and deepen the understanding of the modified proteins with an experimental approach. The work described herein aims at the design and production of a biomimetic vector containing the single-chain variable domain fragment (scFv) of an anti-HER2 antibody fragment as a targeting motif fused with HIV gp41. Molecular modeling and docking studies were performed to develop the recombinant protein sequence. Subsequently, the DNA plasmid was produced and HEK-293T cells were transfected to evaluate the designed vector. The obtained results demonstrated that the plasmid construction is robust and can be expressed in the selected cell line. The multidisciplinary integrated in silico and experimental strategy adopted for the construction of a recombinant protein which can be used in HER2+-targeted therapy paves the way towards the production of other therapeutic proteins in a more cost-effective way.

This study aims to assess the airborne bioburden of rural and urban Portuguese Primary Health Care Centers (PHCC) using active and passive sampling methods and identify the potential differences in airborne microbiota between both environments. The highest total aerobic mesophilic bacterial load in indoor air were found in the Vaccination Room (448 CFU.m−3) in the Rural PHCC and in the Waiting Room (420 CFU.m−3) for Urban PHCC. The total coliforms contamination level in indoor air was detected only in the Cleaning Supplies Room (4 CFU.m−3) in the Urban PHCC. The most frequent bacteria genera identified was Micrococcus (21% Rural PHCC, 31% Urban PHCC). The surface samples showed a highest total aerobic mesophilic bacterial contamination in the Treatment Room (86 × 103 CFU.m−2) from the Rural PHCC and in the Front Office (200 × 103 CFU.m−2) from the Urban PHCC. The electrostatic dust cloth (EDC) samples showed a highest bacterial load in the Urban PHCC. Total aerobic mesophilic bacterial load in settled dust and in the Heating, Ventilating and Air Conditioning (HVAC) filter samples in the Urban PHCC (8 CFU.g−1 and 6 × 103 CFU.m−2) presented higher values compared with the Rural PHCC (1 CFU.g−1 and 2.5 × 103 CFU.m−2). Urban PHCC presented higher bacterial airborne contamination compared with the Rural PHCC for the majority of sampling sites, and when compared with the indoor air quality (IAQ) Portuguese legislation it was the Rural PHCC in two sampling places who did not comply with the established criteria.

© 2020 Elsevier Ltd. All rights reserved., A wider characterization of indoor air quality during sleep is still lacking in the literature. This study intends to assess bioburden before and after sleeping periods in Portuguese dwellings through active methods (air sampling) coupled with passive methods, such as electrostatic dust cloths (EDC); and investigate associations between before and after sleeping and bioburden. In addition, and driven by the lack of information regarding fungi azole-resistance in Portuguese dwellings, a screening with supplemented media was also performed. The most prevalent genera of airborne bacteria identified in the indoor air of the bedrooms were Micrococcus (41%), Staphylococcus (15%) and Neisseria (9%). The major indoor bacterial species isolated in all ten studied bedrooms were Micrococcus luteus (30%), Staphylococcus aureus (13%) and Micrococcus varians (11%). Our results highlight that our bodies are the source of the majority of the bacteria found in the indoor air of our homes. Regarding air fungal contamination, Chrysosporium spp. presented the highest prevalence both in after the sleeping period (40.8%) and before the sleeping period (28.8%) followed by Penicillium spp. (23.47% morning; 23.6% night) and Chrysonilia spp. (12.4% morning; 20.3% night). Several Aspergillus sections were identified in air and EDC samples. However, none of the fungal species/strains (Aspergillus sections Fumigati, Flavi, Nidulantes and Circumdati) were amplified by qPCR in the analyzed EDC. The correlations observed suggest reduced susceptibility to antifungal drugs of some fungal species found in sleeping environments. Toxigenic fungal species and indicators of harmful fungal contamination were observed in sleeping environments., This work was supported by Instituto Politécnico de Lisboa, Lisbon, Portugal for funding the Project "E2SleepeThe impact of environmental exposure on sleep” (IPL/2017/E2SLEEP/ESTeSL/711030) and the Project “Occupational exposure of ambulance drivers to bioburden” (IPL/2020/BIO-AmbuDrivers_ESTeSL). H&TRC authors gratefully acknowledge the FCT/MCTES national support through the UIDB/05608/2020 and UIDP/05608/2020. N. Canha also acknowledges the national FCT funding for his Post doc grant (SFRH/BPD/102944/2014) and his contract ISD-ID (IST-ID/098/2018). The FCT support is also acknowledged by C2TN/IST (UIDB/04349/2020+UIDP/04349/2020) and CESAM (UIDB/50017/2020+UIDP/50017/2020). This study also had the support of LIFE Index-Air project (LIFE15 ENV/PT/000674). It reflects only the view of the authors and EASME is not responsible for any use that may be made of the information it contains.

Electrons with energies of 300 keV or lower have the potential to decontaminate the surfaces of various types of food products with minimal loss of quality. The aim of the present work was to determine the thickness of the layer inhabited by microorganisms. The food samples tested were black and white pepper irradiated with 200 keV, 230 keV, 300 keV and 9 MeV beams of electron energy. To determine the depth from the surface which can be inhabited by microorganisms two approaches were tested. The methods used were based on the application of different microbiological recovery techniques and the microbial effectiveness of the irradiation process depending on the energy of the electron beam. It was observed that the layer which microorganisms may contaminate differed for the tested samples it was estimated as being below 100 μm thick for white pepper and about 200 μm for black pepper. The penetration ability was significant in experiments performed, and as a result the electron beam at the lowest levels tested (200 and 230 keV) was found to be insufficient to effectively decontaminate the black pepper samples. The beam of energy 300 keV was found to have a similar microbial inactivation effect as the high energy electron beam (9 MeV).