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Introduction: Autonomic dysfunction is a common condition after brain injury (BI) and is related to the ability to ensure cerebral self-regulation, which, when present, ensures cerebral blood flow adequate to the person's needs. Rehabilitation training on an inclined plane is related to sensory, autonomic and balance recovery. In this sense, we intend to evaluate cerebrovascular adaptation in people in critical condition (PCS) after BI undergoing standing training using an inclined plane. Methodology: Retrospective multi-case study, carried out in a Neurocritical Care Unit. Sample consisting of (n=7). A rehabilitation program was applied using an inclined plane, with elevation of the headrest up to 60° in two moments. The variables collected were: Glasgow Coma Scale, blood pressure, heart rate (HR) and cerebral oximetry (CO). Through retrospective analysis, the following were studied: heart rate variability (HRV) and baroreflex sensitivity (BS). Results: Median ECG 11, SAPS II 31±23 and average stay of 67±26 days. In the supine position, HRV and BS were decreased. Statistically significant differences were identified in HR during elevation of the headrest up to 60° in the first session (p=0.03). No statistical difference was identified in the calculated variables (p>0.05). Discussion: Compared to the first training, there was an improvement in the regulation of sympathetic-vagal balance observed in the second training, through higher CO values. Conclusion: Standing training using an inclined plane proved to be an effective and safe option in the rehabilitation of PCS after BI, as there was an improvement in cerebrovascular adaptation from the first to the second training., Introducción: La disfunción autonómica es una condición común después de una lesión cerebral (LE) y está relacionada con la capacidad de asegurar la autorregulación cerebral, que, cuando está presente, asegura un flujo sanguíneo cerebral adecuado a las necesidades de la persona. El entrenamiento de rehabilitación en plano inclinado está relacionado con la recuperación sensorial, autonómica y del equilibrio. En este sentido, pretendemos evaluar la adaptación cerebrovascular en personas en estado crítico (PEC) tras LE sometidos a entrenamiento de pie mediante un plano inclinado. Metodología: Estudio retrospectivo de múltiples casos, realizado en una Unidad de Cuidados Neurocríticos. Muestra compuesta por (n=7). Se aplicó un programa de rehabilitación mediante plano inclinado, con elevación del reposacabezas hasta 60° en dos momentos. Las variables recogidas fueron: escala de coma de Glasgow, presión arterial, frecuencia cardíaca (FC) y oximetría cerebral (OC). Mediante análisis retrospectivo se estudió: la variabilidad de la frecuencia cardíaca (VFC) y la sensibilidad barorrefleja (SB). Resultados: Mediana de ECG 11, SAPS II 31±23 y estancia media de 67±26 días. En posición supina, la VFC y la SB disminuyeron. Se identificaron diferencias estadísticamente significativas en la FC durante la elevación del reposacabezas hasta 60° en la primera sesión (p=0,03). No se identificó diferencia estadística en las variables calculadas (p>0,05). Discusión: En comparación con el primer entrenamiento, se observó una mejora en la regulación del equilibrio simpático-vagal en el segundo entrenamiento, a través de valores más altos de CO. Conclusión: El entrenamiento de pie utilizando un plano inclinado demostró ser una opción eficaz y segura en la rehabilitación de la CEP después de LE, ya que hubo una mejora en la adaptación cerebrovascular del primer al segundo entrenamiento., Introdução: A disfunção autonómica é uma condição frequente pós lesão encefálica (LE) e indica a capacidade de assegurar a autorregulação cerebral, que, quando presente, assegura o fluxo sanguíneo cerebral adequado às necessidades da pessoa. O treino de reabilitação em plano inclinado relaciona-se com a recuperação sensorial, autonómica e de equilíbrio. Pretende-se avaliar a adaptação cerebrovascular na pessoa em situação crítica (PSC) pós LE submetida ao treino de ortostatismo através de plano inclinado. Metodologia: Estudo multicaso retrospetivo, realizado numa Unidade de Neurocríticos. Amostra constituída por (n=7). Aplicou-se um programa de reabilitação utilizando o plano inclinado, com elevação da cabeceira até 60° em dois momentos. As variáveis recolhidas foram: Escala de Coma de Glasgow, pressão arterial, frequência cardíaca (FC) e oximetria cerebral (CO). Através de análise retrospetiva estudaram-se: variabilidade da frequência cardíaca (VFC) e sensibilidade do barorreflexo (SB). Resultados: Mediana ECG 11, SAPS II 31±23 e estadia média de 67±26 dias. Em posição supina, a VFC e a SB foram diminuídas. Identificaram-se diferenças estatisticamente significativas na FC durante a elevação da cabeceira até 60° na primeira sessão (p=0,03). Não foi identificada nenhuma diferença estatística nas variáveis calculadas (p>0,05). Discussão: Comparativamente com o primeiro treino, verificou-se melhoria na regulação do equilíbrio simpático-vagal observado no segundo treino, através dos valores de CO mais elevados. Conclusão: O treino de ortostatismo com recurso a plano inclinado demonstrou ser uma opção segura na reabilitação da PSC pós LE, pois verificou-se melhoria na adaptação cerebrovascular do primeiro para o segundo treino.

Trioza erytreae is one natural psyllid vector of Candidatus liberibacter, the causal agent of the citrus greening disease (HLB). Since its introduction in 2014 into the Iberian Peninsula, T. erytreae was able to spread continuously toward southern coastal regions of Portugal and northern coastal regions of the Cantabric sea in Spain. Identifying key areas of potential colonization by the psyllid vector is crucial to anticipate derived problems of establishing the disease. This work aimed to find potential areas that could be colonized by T. erytreae throughout the Iberian Peninsula using 1 km spatial data resolution, a physical barrier, long-distance dispersal events (LDEs), and 30 years of simulation. The bioclimatic suitability of Citrus spp. was used as a surrogate of its spatial distribution and a measure of habitat invasibility. Two localities, one in Portugal and one in Spain, were considered as initial points of the spread of T. erytreae. Four spread scenarios were initially simulated, corresponding to very low, low, medium, and high spread. An altitude of 400 m was included in the model as a physical barrier, and cells up to 500 km apart were allowed to be colonized due to LDEs. In this work, three key risk areas were identified, the citrus-growing areas of Setúbal (Portugal) and Huelva (Spain), and the potential corridors that may connect Guipúzcoa, where T. erytreae is already present, with the Ebro valley (Spain). Future modeling and simulation studies should include biological constraints such as the effect of parasitoids on the population of T. erytreae (e.g., Tamarixia dryi) and other barriers or corridors.

Philaenus spumarius is a vector of Xylella fastidiosa, one of the most dangerous plants pathogenic bacteria worldwide. There is currently no control measure against this pathogen. Thus, the development of vector control strategies, like generalist predators, such as spiders, could be essential to limit the spread of this vector-borne pathogen. In this study, a polymerase chain reaction (PCR)-based approach was developed to principally detect DNA of P. spumarius in the spider’s gut. Accordingly, 20 primer pairs, targeting the mitochondrial cytochrome oxidase I (COI) and cytochrome b (cytB) genes, were tested for specificity, sensitivity, and efficiency in detecting P. spumarius DNA. Overall, two primer sets, targeting COI gene (COI_Ph71F/COI_Ph941R) and the cytB gene (cytB_Ph85F/cytB_Ph635R), showed the highest specificity and sensitivity, being able to amplify 870 pb and 550 bp fragments, respectively, with P. spumarius DNA concentrations 100-fold lower than that of the DNA of non-target species. Among these two primer sets, the cytB_Ph85F/cytB_Ph635R was able to detect P. spumarius in the spider Xysticus acerbus, reaching 50% detection success 82 h after feeding. The feasibility of this primer set to detect predation of P. spumarius by spiders was confirmed in the field, where 20% of the collected spiders presented positive amplifications.

[BACKGROUND] The citrus greening disease or Huanglongbing (HLB) is the most devastating disease of citrus crops. Trioza erytreae is a vector of HLB. Since its introduction in Europe, the insect reached the northern region of Spain and the southern region of Portugal, threatening relevant citrus production areas. Limiting the spread of HLB vectors is mandatory to prevent this disease. In this work, we assessed the effect of kaolin, a white mineral clay, on the landing, settling behavior and feeding behavior of Trioza erytreae on lemon plants., [RESULTS] After kaolin application, the number of plants on which the insect was found was significantly lower than on untreated plants in the laboratory and in the field. Moreover, there were significantly fewer T. erytreae and a shorter duration of phloem-related events on kaolin-treated than untreated plants., [CONCLUSION] The use of kaolin could be a suitable and efficient tool for inclusion into integrated pest management programs or organic production to reduce populations of T. erytreae and subsequently limit the spread of HLB in citrus crops.

Landscape and crop management are important concepts for conservation biological control as they can influence the abundance of natural enemies. In this work we accomplished a multi-scale study focused on the effect of landscape structure and crop management on the olive moth, Prays oleae (Bernard), an important pest of the olive tree (Olea europaea L.). The olive moth was collected in different olive groves managed under organic farming and integrated production and surrounded by different landscape structures. Generalized additive mixed models were used to analyze the response of the pest to (i) landscape composition and configuration indices, (ii) the management system and (iii) pesticides applications at different scales. Results indicated that the landscape composition, through the effect of the Simpson's diversity index, negatively influenced P. oleae abundance and that the effect was evident at larger scales. Also, the landscape configuration negatively affected P. oleae at larger scales. However, neither the crop management system nor the pesticide applications affected P. oleae abundance. This study emphasizes that the preservation or implementation of diverse and complex landscapes can contribute to maintain lower population levels of P. oleae when compared with homogenous landscape areas.

The permeability of the crop surroundings to pests and the landscape structure can influence pest dispersal between crop patches as well as its abundance within the crop. In this work, we analyzed the dispersal of the olive moth Prays oleae (Bernard) throughout the olive grove surroundings and their abundance within the crop following three approaches: (i) pollen grains settled on bodies of olive moths collected in olive groves were identified and compared with flora occurring on the surrounding patches; (ii) the capability of P. oleae males to penetrate non-crop patches was analyzed (iii) the effect of the landscape structure on the abundance of the three generations of the olive moth was studied. Pollen grains of scrubs and other trees occurring in the crop surroundings, such as Cistus sp., Quercus sp., Juniperus-type or Pinaceae were identified on P. oleae bodies suggesting that P. oleae penetrates into non-crop habitats. Additionally, woody and, to a lesser degree, herbaceous patches, did not constitute barriers for P. oleae. Finally, more complex and heterogeneous patches presented lower numbers of captures of P. oleae. These results give new insights into the movements of the olive moth in the olive grove surroundings and suggest that the management of non-crop areas could influence this pest abundance.

Chrysoperla carnea s.l. (Stephens) larvae are important natural enemies in agroecosystems. However, adults feed on honeydew excreted by hemipterans, and pollen and nectar from flowering plants. Pollen is essential for egg production, but to our knowledge, the pollen diet of C. carnea in perennial crops has never been addressed. In this work, the objective was to study the diversity and potential selection of pollen types consumed by C. carnea in the olive grove agroecosystem. For this study, C. carnea adults were captured from April to December of 2012 and 2013, and simultaneously, inventories of the plant diversity were obtained in olive orchards and adjacent scrubland and herbaceous patches. The pollen types contained in C. carnea guts were identified by microscopy and compared with the pollen types in the environment using Jacobs`s second selection index. The results indicated that (i) C. carnea females and males captured in the olive tree canopy visited scrub and herbaceous vegetation patches; (ii) they fed on different anemophilous and entomophilous pollen types from tree and scrub (Olea europaea, Fabaceae, Pinaceae, Cistaceae or Ericaceae) and herbaceous (Asteraceae, Apicaceae, Brassicaceae, Poaceae, Rumex type or Plantago type) strata; and (iii) adults fed not only on flowers but also on pollen settled on vegetation surfaces. Here, we demonstrated that C. carnea could benefit not only from the diversity of entomophilous pollen during the plant flowering periods but also from other pollens that could remain in the environment in different periods. This has important implications for the management of the floral diversity adjacent to the crop.

The bacterium Xylella fastidiosa (Xf) is an invasive insect-borne pathogen, which causes lethal diseases to important crops including olives, citrus, almonds and grapes as well as numerous forest, ornamental, and uncultivated plants. Outbreaks of Xf-related plant diseases are currently occurring in the Mediterranean region, causing substantial losses to various agricultural sectors. Several models have recently been published to identify which regions are at highest risk in Europe; however, such models did not consider the insect vectors, which constitute the key driver of short-range Xf spread. We fitted bioclimatic species distribution models to depict the macroclimatic preferences of the meadow spittlebug Philaenus spumarius L. (1978) (Hemiptera: Aphrophoridae), the major epidemiologically relevant vector currently responsible for Xf spread in the Europe. Many regions of Western Europe and Mediterranean basin are predicted by models as highly climatically suitable for this vector, including all regions where severe Xf have occurred so far. Conversely, the driest and warmest areas of the Mediterranean basin are predicted as little suitable for P. spumarius. Models forecast that agricultural-important parts of the southern Mediterranean area might experience a substantial decrease in climatic suitability for P. spumarius by the period 2040–2060. Areas predicted as highly suitable just for the bacterium but not optimal for this vector are apparently still free of severe Xf outbreaks, suggesting that climate tolerances of P. spumarius might partly explain the current spatial pattern of Xf outbreaks in Europe and should always be considered in further risk assessments.

The bacterium Xylella fastidiosa (Xf) is an invasive insect-borne pathogen, which causes lethal diseases to important crops including olives, citrus, almonds and grapes as well as numerous forest, ornamental, and uncultivated plants. Outbreaks of Xf-related plant diseases are currently occurring in the Mediterranean region, causing substantial losses to various agricultural sectors. Several models have recently been published to identify which regions are at highest risk in Europe; however, such models did not consider the insect vectors, which constitute the key driver of short-range Xf spread. We fitted bioclimatic species distribution models to depict the macroclimatic preferences of the meadow spittlebug Philaenus spumarius L. (1978) (Hemiptera: Aphrophoridae), the major epidemiologically relevant vector currently responsible for Xf spread in the Europe. Many regions of Western Europe and Mediterranean basin are predicted by models as highly climatically suitable for this vector, including all regions where severe Xf have occurred so far. Conversely, the driest and warmest areas of the Mediterranean basin are predicted as little suitable for P. spumarius. Models forecast that agricultural-important parts of the southern Mediterranean area might experience a substantial decrease in climatic suitability for P. spumarius by the period 2040–2060. Areas predicted as highly suitable just for the bacterium but not optimal for this vector are apparently still free of severe Xf outbreaks, suggesting that climate tolerances of P. spumarius might partly explain the current spatial pattern of Xf outbreaks in Europe and should always be considered in further risk assessments.

Landscape and crop management are important concepts for conservation biological control as they can influence the abundance of natural enemies. In this work we accomplished a multi-scale study focused on the effect of landscape structure and crop management on the olive moth, Prays oleae (Bernard), an important pest of the olive tree (Olea europaea L.). The olive moth was collected in different olive groves managed under organic farming and integrated production and surrounded by different landscape structures. Generalized additive mixed models were used to analyze the response of the pest to (i) landscape composition and configuration indices, (ii) the management system and (iii) pesticides applications at different scales. Results indicated that the landscape composition, through the effect of the Simpson’s diversity index, negatively influenced P. oleae abundance and that the effect was evident at larger scales. Also, the landscape configuration negatively affected P. oleae at larger scales. However, neither the crop management system nor the pesticide applications affected P. oleae abundance. This study emphasizes that the preservation or implementation of diverse and complex landscapes can contribute to maintain lower population levels of P. oleae when compared with homogenous landscape areas., Portuguese Foundation of Science and Technology (FCT, Portugal), Centro de Investigação de Montanha (CIMO, Portugal), Depto. de Biodiversidad, Ecología y Evolución, Fac. de Ciencias Biológicas, TRUE, pub

Assessing the potential of spread of an introduced crop pest in a new country is crucial to anticipating its effects on crop production and deriving phytosanitary management toward reducing potential negative effects. Citrus production represents a key agricultural activity throughout the Mediterranean basin. The African citrus psyllid Trioza erytreae (del Guercio, 1918) (Hemiptera: Triozidae) is a natural vector of Candidatusliberibacter spp., the causal agent of the harmful disease huanglongbing (HLB) or “citrus greening disease”. In continental Europe, T. erytreae was detected for the first time in northwestern Spain in 2014. Pest risk analysis (PRA) approaches, such as modeling, consider both time and space components to predict the potential distribution of pests in a given region. In this work, we aim to parameterize a model able to predict the expected spread of T. erytreae in the Iberian Peninsula using three types of PRA models. The kernel model with two hypothetical entry points accurately predicted the distribution of T. erytreae with respect to latitude. This model should be further refined and validated to support decision-makers in the adoption of timely and successful management and regulatory measures against the spread of T. erytreae to other citrus-producing areas in Europe.

The olive grove is a key landscape across the Mediterranean basin. This agroecosystem is threatened by Xylella fastidiosa, the causal agent of the olive tree quick decline syndrome, Philaenus spumarius being the main vector. A way to limit pest populations relies on the use of biological control agents such as arthropods. Among them, spiders are generalist predators with different hunting strategies that feed mostly on insects and can contribute to limit pests. In this work, field and laboratory data were used to provide a protocol aiming to facilitate the selection of species of spiders among different guilds that could represent potential natural enemies of P. spumarius. Sampling of spiders was conducted in olive groves in northeastern Portugal. Two species, namely the orb-weaver Araniella cucurbitina and the ambusher Synema globosum, were selected according to the dominant guilds of spiders inhabiting the olive crop. We tested the differences of potential predatory efficiency using classical functional response tests with P. spumarius as prey. A type-II functional response was found for A. cucurbitina, whereas a type-I response was found for S. globosum. This difference uncovers a different potential efficiency among the two species as natural enemies of P. spumarius with relevant implications at high prey density in the field. A conceptual workflow to follow the fieldwork and selection of species for further work (i.e., laboratory assays) is provided and discussed. Standardized methods regarding the assessment of the suitability and efficiency of potential natural enemies are essential for the integration of results at different geographical extents and crops. Selecting functional counterparts such as different species of predators occurring at different locations that use the same prey (e.g., a pest) in the same way (e.g., hunting strategy) would facilitate developing biological control schemes.

[EN] The olive processing industry has till date been dominated by a small group of cultivars, leading to the possibility of some olive cultivars becoming extinct in the near future. In this study, we determined the composition of some chemical components in the olive oils from 31 minor olive cultivars of the Valencian Community. Our main aim was to identify suitable cultivars, which could produce differentiated olive oils, thus aiming towards their valorization. The average oil content of minor olive cultivars was found to be good, with some of them reporting approximately 60% (dry basis). On average, the total phenolic content was 229mg kg(-1), with cv. Mas Blanc reporting the highest content (570mg kg(-1)). Among the various tocopherols found in olives, -tocopherol was the main constituent, with a maximum concentration of 290.6mg kg(-1). Linoleic acid was the main polyunsaturated fatty acid and varied between 3.4% (cv. Del Pomet) and 16.9% (cv. Blanqueta Enguera). Special attention needs to be paid to the composition of sterols, since some olive oils exceeded the limits established for some sterols by the current European legislation. Some of the cultivars studied were highly productive, and originated differentiated olive oils with a rich composition of antioxidants and essential fatty acids. In some cases, these beneficial compounds were higher than those of commercial oils obtained from the most common cultivars worldwide. These results could contribute to the commercial exploitation of some of the studied cultivars.

[EN] During the last ten years we have assisted to the consolidation of the almond crop that has remarkably increased its cultivation area causing a high demand for both plants and products related to growth stimulation. Accordingly, in the present work we aim to study the changes produced by the contribution of two biostimulants (humic and fulvic acids or aminoacids) on the properties of almond tree rootstocks. This kind of studies are of interest to the nursery cultivation industry where rapid growth of trees and good adaptation to their cultivation environment are required. Plants' radicular and vegetative systems responded differently according to the rootstock selection. The fastest and vigorous vegetative development was observed in GN rootstock whereas GF 677 showed the greatest number of main roots and RP-R of secondary roots. Differences on antioxidant activity and phenol content have also been found between rootstocks. All the tested samples were found to have a high antioxidant power and a high phenol content but GN stood out in this regard over the other rootstocks under study. The efficiency of the biostimulants applied has been verified. Both biostimulants promoted the development of the aerial part of the trees but biostimulant 2 did it to a greater extent. Biostimulant 1 was able to duplicate the number of main roots in RP-R and during the first year of study, biostimulant 2 originated an increase of the weight of the root system by 26.44% for RP-R, 16.93% for GF 677 and 48.00% for GN. In view of these results, synthetic chemical fertilizers can be at least partially replaced by biostimulants.

[EN] The olive processing industry has till date been dominated by a small group of cultivars, leading to the possibility of some olive cultivars becoming extinct in the near future. In this study, we determined the composition of some chemical components in the olive oils from 31 minor olive cultivars of the Valencian Community. Our main aim was to identify suitable cultivars, which could produce differentiated olive oils, thus aiming towards their valorization. The average oil content of minor olive cultivars was found to be good, with some of them reporting approximately 60% (dry basis). On average, the total phenolic content was 229mg kg(-1), with cv. Mas Blanc reporting the highest content (570mg kg(-1)). Among the various tocopherols found in olives, -tocopherol was the main constituent, with a maximum concentration of 290.6mg kg(-1). Linoleic acid was the main polyunsaturated fatty acid and varied between 3.4% (cv. Del Pomet) and 16.9% (cv. Blanqueta Enguera). Special attention needs to be paid to the composition of sterols, since some olive oils exceeded the limits established for some sterols by the current European legislation. Some of the cultivars studied were highly productive, and originated differentiated olive oils with a rich composition of antioxidants and essential fatty acids. In some cases, these beneficial compounds were higher than those of commercial oils obtained from the most common cultivars worldwide. These results could contribute to the commercial exploitation of some of the studied cultivars.

[EN] During the last ten years we have assisted to the consolidation of the almond crop that has remarkably increased its cultivation area causing a high demand for both plants and products related to growth stimulation. Accordingly, in the present work we aim to study the changes produced by the contribution of two biostimulants (humic and fulvic acids or aminoacids) on the properties of almond tree rootstocks. This kind of studies are of interest to the nursery cultivation industry where rapid growth of trees and good adaptation to their cultivation environment are required. Plants' radicular and vegetative systems responded differently according to the rootstock selection. The fastest and vigorous vegetative development was observed in GN rootstock whereas GF 677 showed the greatest number of main roots and RP-R of secondary roots. Differences on antioxidant activity and phenol content have also been found between rootstocks. All the tested samples were found to have a high antioxidant power and a high phenol content but GN stood out in this regard over the other rootstocks under study. The efficiency of the biostimulants applied has been verified. Both biostimulants promoted the development of the aerial part of the trees but biostimulant 2 did it to a greater extent. Biostimulant 1 was able to duplicate the number of main roots in RP-R and during the first year of study, biostimulant 2 originated an increase of the weight of the root system by 26.44% for RP-R, 16.93% for GF 677 and 48.00% for GN. In view of these results, synthetic chemical fertilizers can be at least partially replaced by biostimulants.

Introduction: Pansies (Viola×wittrockiana) are a rich source of natural antioxidants with beneficial effects on human health. Objetives: The aim of our study was to investigate solvents’ influence (water, methanol, water:acetone (6:4, v/v)) on the extraction of bioactive compounds and antioxidant activity of pansies extracts. Methods: The bioactive compounds analyzed were the following: flavonoids, hydrolysable tannins and monomeric anthocyanins, as well as total phenols by the total reducing capacity assay (TRC). The antioxidant activity was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and reducing power assays. A Principal Component Analysis (PCA) was performed to differentiate pansies extracts. Results: The solvents that yielded extracts with the highest contents of hydrolysable tannins and TRC were methanol and water:acetone (6:4, v/v). To extract the highest contents of monomeric anthocyanins, methanol should be used (5.93 mg Cy 3-glu/g flower, d.w), while for flavonoids, water:acetone (6:4, v/v) was the preferred yielding an extract with 115 mg QE/g flower d.w. Water turned out to be the least effective solvent, giving extracts with the lowest antioxidant activity. In addition, methanol or water:acetone extracts were clearly distinguished from aqueous ones through a PCA analysis. Conclusions: Our results show that the bioactive compounds and antioxidant activity of pansies’ extracts are affected by the solvent used., Introdução: Amores-perfeitos (Viola×wittrockiana) são uma fonte rica de antioxidantes naturais com efeitos benéficos para a saúde humana. Objetivos: O objetivo do presente estudo foi investigar a influência dos solventes (água, metanol e água:acetona (6:4, v/v)) na extracção de compostos bioactivos e actividade antioxidante de extractos de amores-perfeitos. Métodos: Os compostos bioactivos analisados foram os seguintes: flavonóides, taninos hidrolisáveis e antocianinas monoméricas, bem como, os fenóis totais através da avaliação da capacidade redutora total. A actividade antioxidante foi avaliada pelos métodos de sequestro do radical livre 2,2-difenil-1-picrilhidrazilo (DPPH) e poder redutor. Uma análise de componentes principais (PCA) foi realizada para diferenciar os extractos dos amores-perfeitos. Resultados: Os solventes que produziram extractos com os maiores teores de taninos hidrolisáveis e capacidade redutora total foram os de metanol e água: acetona (6:4, v/v). Para extrair os maiores teores de antocianinas monoméricas deve-se usar metanol (5.93 mg Cy 3-glu/ g de massa seca), enquanto que para extrair flavonóides, a água:acetona (6:4, v/v) é preferível (115 mg QE/g de massa seca). A água mostrou ser o solvente menos eficaz, obtendo-se extractos com a menor actividade antioxidante. Para além disso, os extractos de metanol e água:acetona foram claramente separados dos aquosos através da análise por PCA. Conclusões: Os resultados mostraram que a extracção de compostos bioactivos e a actividade antioxidante de flores de amores-perfeitos são influenciadas pelo solvente usado., Introducción: Los pensamientos (Viola x wittrockiana) son una rica fuente de antioxidantes naturales con efectos beneficiosos sobre la salud humana. Objetivos: El objetivo del presente estudio fue evaluar la influencia de los solventes (agua, metanol y agua:acetona (6:4, v/v) en la extracción de compuestos bioactivos y actividad antioxidante de extractos de pensamientos. Métodos: Los compuestos bioactivos analizados fueron flavonoides, taninos hidrolizados y antocianinas monoméricas. También se analizaron los fenoles totales por el ensayo de capacidad reductora total. La actividad antioxidante fue evaluada por los métodos: secuestro del radical libre 2,2-difenil-1-picrilhidrazilo (DPPH) y poder reductor. El análisis de componentes principales (PCA) fue realizado para diferenciar los extractos de los pensamientos. Resultados: Los solventes que produjeron extractos con los mayores contenidos de taninos hidrolizados y capacidad reductora total fueron el metanol y agua:acetona (6:4, v/v). Los extractos con mayores contenidos de antocianinas monoméricas se obtuvieron utilizando metanol (5.93 mg Cy 3-glu/g peso seco). Para los flavonoides el agua: acetona (6:4, v / v) fue el solvente que hay dado los mejores valores de extracción (115 mg QE / g peso seco). El agua fue el solvente menos eficaz, obteniéndo se extractos con menor actividad antioxidante. Además, los extractos de metanol y agua:acetona se separaron claramente de los acuosos mediante el análisis por PCA. Conclusións: Los resultados mostraron que la extracción de compuestos bioactivos y la actividad antioxidante de los pensamientos son influenciadas por el solvente utilizado.

Many syrphid larvae are predators of aphids and early stages of moths and psyllids, being potential control agents of some olive pests. However, adults need pollen and nectar for energy and reproduction. An increase of plant resources biodiversity in heterogeneous landscape could benefit these natural enemies. Thus, our goal of was to study the syrphid biodiversity and their pollen sources in olive agroecosystems from the northeast of Portugal during spring, that coincides with the availability of pests. For that, syrphids were captured in three not-tilled olive groves and two surrounding field areas (an herbaceous vegetation patch and a scrubland) next to each olive grove. Simultaneously, flowering plant inventories were carried out. Syrphids and the pollen contained in the guts of the most abundant species were identified. Thereafter pollen in guts was compared with pollen in the environment to evaluate a potential pollen selection. The most abundant species were Sphaerophoria scripta and Melanostoma mellinum. During the spring syrphids did not consume all the occurring plant species but did not actively selected pollen types. Results indicated that Asteraceae, Plantaginaceae, Caryophyllaceae, Echium type, Rumex type and Jasione type are important food sources for M. mellinum and S. scripta. These results will allow managing ground cover vegetation more efficiently in order to conserve syrphids in the olive agroecosystem

The species Olea europaea L. is constituted by two varieties, the cultivated one - the olive tree, O. europaea subsp. europaea var. europaea; and the wild form - the wild olive tree or oleaster, O. europaea subsp. europaea var. sylvestris. These varieties were differentiated during the domestication process, and from the cultivated form a high number of cultivars evolved over the years. In the last decades, due to the crop intensification a few number of well adapted or productive cultivars have been used in the new olive plantations, leading to the abandonment of minor cultivars and to a loss of the olive genetic heritage. Nevertheless, in the last years, a niche of more informed and demanding consumers appeared, searching for olive oils that combine their richness in health promoters, like polyphenols, tocopherols and sterols, with a differentiated sensory profile. In this context, the general objective of this work was to characterize genetically and morphologically oleander populations and centenarian olive trees from the northeast of Portugal, as well as, to chemically and sensory evaluate the extracted oils, aiming the selection of plant specimens with the purpose of future valorization. Three oleander populations from Alijó, Moncorvo and Vila Nova de Foz Côa (VNFC) regions and 28 specimens of centenarian olive trees, grown in Mirandela region (Suçães), some of them from known cultivars (cvs. Lentisca, Madural, Rebolã, Redondal, Verdeal and Verdeal Transmontana) were characterized. The comparison of genetic diversity, structure and phylogenetic relationships within and among the oleaster and centenarian plants were studied using microsatellite markers. High genetic diversity was observed in both oleaster and centenarian plants, with no differences between them in the amount of genetic diversity. Population structure analysis suggests genetic differentiation between the studied varieties. The oils of the three oleaster populations were evaluated considering the

The black scale, Saissetia oleae (Olivier), is considered a secondary pest of olive tree in Portugal, where it is attacked by a complex of natural enemies that control its population. Parasitoids of genus Metaphycus (Hymenoptera: Encyrtidae) and Coccophagus (Hymenoptera: Aphelinidae) are among the main natural enemies. In this thesis, the several aspects of the successful natural biological control of this scale are analyzed in the field and laboratory. In detail, we analyzed: (i) whether the facultative autoparasitoid Coccophagus lycimnia (Walker) can adversely affect the population densities of Metaphycus spp. parasitoids and disrupt the biological control of S. oleae; (ii) the effect of ant-exclusion on the biological control of S. oleae and its parasitoid complex; and (iii) the effect of the carbohydrate sources available in the field (nectar and honeydew) on the survival of parasitoids of both genus. For the development of the first objective, the population densities of the scale, the primary parasitoids of the genus Metaphycus and the autoparasitoid C. lycimnia were recorded for three consecutive seasons in 28 olive groves. The populations of the parasitoids followed opposite patterns. At the end of the life cycle of the S. oleae (May), the number of Metaphycus spp. observed was practically nil whereas the number of C. lycimnia was five times greater than at the beginning of the assay. In September and November, Metaphycus spp. parasitoids parasitized smaller scales than C. lycimnia, demonstrating that Metaphycus spp. parasitoids are superior at exploitative competition. Metaphycus spp. and C. lycimnia sex ratio was similar in September, but in November the sexual proportion of Metaphycus spp. was higher than that of C. lycimnia. The facultative autoparasitoid C. lycimnia tended to lay male eggs on Metaphycus spp. females. The density of S. oleae decreased from September to May, when the ratio S. oleae-C. lycimnia reached 2:1. Altogether, these results demons

The species Olea europaea L. is constituted by two varieties, the cultivated one - the olive tree, O. europaea subsp. europaea var. europaea; and the wild form - the wild olive tree or oleaster, O. europaea subsp. europaea var. sylvestris. These varieties were differentiated during the domestication process, and from the cultivated form a high number of cultivars evolved over the years. In the last decades, due to the crop intensification a few number of well adapted or productive cultivars have been used in the new olive plantations, leading to the abandonment of minor cultivars and to a loss of the olive genetic heritage. Nevertheless, in the last years, a niche of more informed and demanding consumers appeared, searching for olive oils that combine their richness in health promoters, like polyphenols, tocopherols and sterols, with a differentiated sensory profile. In this context, the general objective of this work was to characterize genetically and morphologically oleander populations and centenarian olive trees from the northeast of Portugal, as well as, to chemically and sensory evaluate the extracted oils, aiming the selection of plant specimens with the purpose of future valorization. Three oleander populations from Alijó, Moncorvo and Vila Nova de Foz Côa (VNFC) regions and 28 specimens of centenarian olive trees, grown in Mirandela region (Suçães), some of them from known cultivars (cvs. Lentisca, Madural, Rebolã, Redondal, Verdeal and Verdeal Transmontana) were characterized. The comparison of genetic diversity, structure and phylogenetic relationships within and among the oleaster and centenarian plants were studied using microsatellite markers. High genetic diversity was observed in both oleaster and centenarian plants, with no differences between them in the amount of genetic diversity. Population structure analysis suggests genetic differentiation between the studied varieties. The oils of the three oleaster populations were evaluated considering the

The black scale, Saissetia oleae (Olivier), is considered a secondary pest of olive tree in Portugal, where it is attacked by a complex of natural enemies that control its population. Parasitoids of genus Metaphycus (Hymenoptera: Encyrtidae) and Coccophagus (Hymenoptera: Aphelinidae) are among the main natural enemies. In this thesis, the several aspects of the successful natural biological control of this scale are analyzed in the field and laboratory. In detail, we analyzed: (i) whether the facultative autoparasitoid Coccophagus lycimnia (Walker) can adversely affect the population densities of Metaphycus spp. parasitoids and disrupt the biological control of S. oleae; (ii) the effect of ant-exclusion on the biological control of S. oleae and its parasitoid complex; and (iii) the effect of the carbohydrate sources available in the field (nectar and honeydew) on the survival of parasitoids of both genus. For the development of the first objective, the population densities of the scale, the primary parasitoids of the genus Metaphycus and the autoparasitoid C. lycimnia were recorded for three consecutive seasons in 28 olive groves. The populations of the parasitoids followed opposite patterns. At the end of the life cycle of the S. oleae (May), the number of Metaphycus spp. observed was practically nil whereas the number of C. lycimnia was five times greater than at the beginning of the assay. In September and November, Metaphycus spp. parasitoids parasitized smaller scales than C. lycimnia, demonstrating that Metaphycus spp. parasitoids are superior at exploitative competition. Metaphycus spp. and C. lycimnia sex ratio was similar in September, but in November the sexual proportion of Metaphycus spp. was higher than that of C. lycimnia. The facultative autoparasitoid C. lycimnia tended to lay male eggs on Metaphycus spp. females. The density of S. oleae decreased from September to May, when the ratio S. oleae-C. lycimnia reached 2:1. Altogether, these results demons

The effect of the experimental conditions on assessing the global volatile profile of extra virgin olive oils (EVOO) by headspace solid-phase micro-extraction/gas chromatography-mass spectrometry (HS-SPME/GC–MS) is studied to obtain maximization of the total peak areas of the compounds. Response Surface Methodology (RSM) is applied to Arbequina EVOO, and the influence of oil quantity, extraction time, and extraction temperature on the total area and extraction of the major desirable compounds is analyzed. The experimental data are adequately fitted into second-order polynomial models with non-significant lack of fit (p > 0.05) and coefficients of determination (R and R -adjusted) higher than 0.88 and 0.78, respectively. A strong similarity is found between the predicted and experimental values. Furthermore, the surface plots show that the extraction of the volatile compounds is favored with increasing extraction temperature, time, and oil quantity. The extraction conditions to obtain the maximum response of “green” volatile compounds from Arbequina olive oil by HS-SPME placed in 50 mL vials are 4.6 g of oil, 43 min, and 59 C. Practical Applications: The findings of the present work show that specific conditions of HS-SPME/GC–MS affect the extraction of volatile compounds from cv. Arbequina olive oil and establish the optimum extraction parameters to more efficiently determine the global profile of this fraction, taking into account the organoleptic characteristics of the cultivar. The results also contribute to the knowledge of the volatile profile of the monovarietal olive oil cv. Arbequina, which is one of most cultivated and consumed types of olive oil worldwide.

The phytosanitary emergency caused by the spread of Xylella fastidiosa in the Mediterranean has raised demands for a better understanding of the ecology of its presumed and candidate insect vectors. Here, we present the results of a two-year survey carried out in olive groves across southern, eastern and Central Spain and northeastern Portugal. Several sampling methods were tested and compared to select the most appropriate to estimate population levels of potential vectors of X. fastidiosa. The spittlebugs Philaenus spumarius and Neophilaenus campestris (Hemiptera: Aphrophoridae) were the main species associated with olive groves. Both species were widely present on herbaceous ground vegetation within the olive groves; P. spumarius mainly associated with Asteraceae and N. campestris with Poaceae. Due to the patchy distribution of spittlebugs within the olive groves, sweep nets were the most effective and least time-consuming sampling method for the estimation of population size both in the ground cover and tree canopies. Trends in population density showed that spittlebugs can be abundant on ground vegetation but very rare on olive canopies. Spittlebugs disperse in late spring to non-cultivated hosts that act as natural reservoirs. In late fall, adults return to the olive groves for oviposition. However, olive trees may act as transient hosts for spittlebugs and high population densities of these insect vectors should be avoided in areas where X. fastidiosa is present.

[EN] Nine pomegranate cultivars grown in Spain were selected, and their physicochemical (total soluble solids, pH, titratable acidity, maturity index, monomeric anthocyanin pigment, flavonoids, hydrolyzable tannins, and vitamin C) and antioxidant properties and polyphenolic composition of the juices were compared. A total of 53 polyphenols were identified, showing cultivars different profiles. Of all nine cultivars, Katirbasi had the highest contents of flavonoids, hydrolyzable tannins and vitamin C, as well as gallic acid and ellagic acid contents, explaining its high total reducing capacity. Principal component analysis allowed Katirbasi to be differentiated clearly from the others. Other cultivars presented also interesting characteristics such as high monomeric anthocyanin pigment content (CG8 cultivar) and interesting antioxidant activity (Wonderful 2 and CG8 cultivars). CG8 was the cultivar with the highest value of cyanidin-3,5-di-O-glucoside. Thus, this study will assist pomegranate producers in choosing the most suitable cultivar according to its ultimate use.

[EN] Nine pomegranate cultivars grown in Spain were selected, and their physicochemical (total soluble solids, pH, titratable acidity, maturity index, monomeric anthocyanin pigment, flavonoids, hydrolyzable tannins, and vitamin C) and antioxidant properties and polyphenolic composition of the juices were compared. A total of 53 polyphenols were identified, showing cultivars different profiles. Of all nine cultivars, Katirbasi had the highest contents of flavonoids, hydrolyzable tannins and vitamin C, as well as gallic acid and ellagic acid contents, explaining its high total reducing capacity. Principal component analysis allowed Katirbasi to be differentiated clearly from the others. Other cultivars presented also interesting characteristics such as high monomeric anthocyanin pigment content (CG8 cultivar) and interesting antioxidant activity (Wonderful 2 and CG8 cultivars). CG8 was the cultivar with the highest value of cyanidin-3,5-di-O-glucoside. Thus, this study will assist pomegranate producers in choosing the most suitable cultivar according to its ultimate use.

[EN] Nine pomegranate cultivars grown in Spain were selected, and their physicochemical (total soluble solids, pH, titratable acidity, maturity index, monomeric anthocyanin pigment, flavonoids, hydrolyzable tannins, and vitamin C) and antioxidant properties and polyphenolic composition of the juices were compared. A total of 53 polyphenols were identified, showing cultivars different profiles. Of all nine cultivars, Katirbasi had the highest contents of flavonoids, hydrolyzable tannins and vitamin C, as well as gallic acid and ellagic acid contents, explaining its high total reducing capacity. Principal component analysis allowed Katirbasi to be differentiated clearly from the others. Other cultivars presented also interesting characteristics such as high monomeric anthocyanin pigment content (CG8 cultivar) and interesting antioxidant activity (Wonderful 2 and CG8 cultivars). CG8 was the cultivar with the highest value of cyanidin-3,5-di-O-glucoside. Thus, this study will assist pomegranate producers in choosing the most suitable cultivar according to its ultimate use.

Introduction: São Tomé and Príncipe has a wide variety of fruit trees, being most of the fruits consumed in fresh. In certain periods of the year, fruits are available at significant amounts; however, it is not common to use the surplus of those fruits. Thus, this excess may be used in the preparation of other fruit based products (ex. jams), in order to increase product diversity and allow the annual consumption of these seasonal fruits. Objetives: Valorise fruits’ production in S. Tomé and Príncipe. Methods: Preparation of jams of banana, ambarella, guava (dark and light) and papaya, with two levels of sugar, and perform their physico-chemical characterization (color, pH, moisture and acidity). Furthermore, sensory analysis was also done, through preference and acceptability tests. Results: The jams prepared had different colours, acidity values, and moisture and ash contents, demonstrating the possibility of producing different products by changing the formulation. Regarding jams preference, 60% of the consumers preferred the less sweet in the case of banana and guava (light), whereas, over 67% of the panellists preferred the sweetest ambarella jam. Concerning dark guava and papaya, the percentages were similar for both sugar contents. For all attributes analysed (appearance, colour, taste, acidity, sweetness and global evaluation), most of the panellists liked slightly all jams. Conclusion: Jams production in S. Tomé and Príncipe may be a promising activity., Introdução: São Tomé e Príncipe tem uma grande variedade de árvores frutíferas, sendo a maioria dos frutos consumida em fresco. Em certos períodos do ano, os frutos estão disponíveis em quantidades significativas, não sendo comum usar o excedente. Assim, o excedente de frutos pode ser usado na preparação de outros produtos (ex. doces), para aumentar a diversidade de produtos e permitir o consumo anual desses frutos sazonais. Objetivos: Valorizar a produção de frutos de S. Tomé e Príncipe. Métodos: Elaboração de doces de banana, cajá-manga, goiaba (escura e clara) e mamão, com diferentes teores de açúcar, e posteriormente proceder à sua caracterização físico-química (cor, pH, teor de humidade e acidez) e sensorial, através de testes de preferência e aceitabilidade. Resultados: Os doces preparados apresentaram diferentes cores e valores de acidez, humidade e cinza, demonstrando a possibilidade de elaboração de diferentes produtos pela modificação da formulação. Quanto à preferência dos doces, 60% dos consumidores preferiram os menos doces no caso da banana e goiaba (clara), enquanto mais de 67% dos participantes preferiram o doce de cajá- manga mais doce. Quanto à goiaba escura e papaia, as percentagens foram semelhantes para ambos os teores de açúcar. Para todos os atributos analisados (aparência, cor, sabor, acidez, doçura e avaliação global), a maioria dos participantes gostou ligeiramente de todos os doces. Conclusões: A produção de doces em S. Tomé e Príncipe pode ser uma actividade promissora, Introducción: S. Tomé y Príncipe tiene una gran variedad de árboles frutales, siendo la mayoría de los frutos comidos en fresco. En ciertos períodos del año, los frutos están disponibles en cantidades significativas, no siendo común el uso de los excedentes. Por lo tanto, este excedente se puede utilizar en la preparación de otros productos y permitir el consumo anual de la fruta de temporada. Objetivos: Valorar la producción de frutos en S. Tomé y Príncipe. Métodos: Preparación de mermelada de plátano, caja-mango, guayaba (clara y oscura) y papaya con distintos contenidos de azúcar, y proceder à su caracterización físico-química (color, pH, contenido de humedad y acidez) y sensorial, a través de pruebas de preferencia y aceptabilidad. Resultados: Las mermeladas preparadas tenían diferentes colores y valores de acidez, humedad y cenizas, lo que demuestra la posibilidad de producir diferentes productos. En relación à la preferencia de las mermeladas, 60% de los consumidores prefiere la menos dulce en el caso del plátano y guayaba (clara), mientras más del 67% de los participantes prefiere la mermelada de caja-mango más dulce. En cuanto a la guayaba oscura y papaya, los porcentajes fueron similares para ambos niveles de azúcar. Para todos los atributos analizados (aspecto, color, sabor, acidez, dulzor y evaluación global), la mayoría de los participantes les gustó un poco todas las mermeladas. Conclusións: La producción de mermeladas en S. Tomé y Príncipe puede ser una actividad prometedora.

Many syrphid larvae are predators and have an important role as biological control agents of pests in agroecosystems. However, adults feed on non-prey resources such as pollen from flowers. Heterogeneous landscapes can provide syrphids with a great biodiversity of plants and ensure the existence of food resources. This is particularly important during periods of food scarcity, such as autumn, for syrphid species that spend those periods as adults. Nevertheless, the feeding habits of syrphid adults in resource-scarce agroecosystems are poorly known. In this study, the pollen consumption and preferences of Eupeodes corollae (Fabricius) and Episyrphus balteatus (De Geer) were analyzed in olive groves and surrounding herbaceous and woody patches in the autumns of 2012 and 2013 in northeastern Portugal. The guts were dissected, and the pollen types were identified and compared with the ground cover plants in the studied patches. Both species consumed and selected pollen types from herbaceous and woody vegetation that occurred in different patches, indicating that they flew between patches. These results highlight the importance of conserving heterogeneous agricultural landscapes to guarantee the existence of food resources for beneficial insects during periods of scarcity.

In the present work, we investigate how the degree of ripeness (low, low-medium, medium and medium-high) aff ects the physical and compositional changes, as well as antioxidant properties of pomegranate fruit (cv. Mollar de Elche). The skin, pellicle, seed and juice were analysed. The fruit mass increased and the fruit skin became reddish (higher a* and lower h*) as the fruit ripening progressed. The lowest concentrations of fl avonoids and hydrolysable tannins were recorded in skin and pellicles at medium-high maturity stage, which explains the decrease in the total phenols and reducing power during ripening of pomegranate. On the contrary, the highest concentration of fl avonoids (165 mg of quercetin equivalents per 100 mL of juice) was determined in the juice at the most advanced ripening stage, concomitant with the highest total phenols (1695 mg of gallic acid equivalents per 100 mL of juice). Higher DPPH scavenging activity and an increase in the reducing power of juice were also observed during ripening. The trend of the above-mentioned properties allowed describing the fruit development and maturity.

[EN] The present study was conducted to determine the major bioactive lipid components of the seed oils of nine pomegranate (Punica granatum L.) cultivars grown in Spain, namely fatty acids, vitamin E and sterol compositions. The seeds yielded oil contents ranging from 4.44% to 13.70% of dry matter and showed high contents of polyunsaturated fatty acids (86.7.2–90.3%). The predominant fatty acid was 9,11, 13-octadeca-trienoic acid (punicic acid), a conjugated linolenic acid characteristic from pomegranate seeds, with contents between 3523 and 10,586 mg/100 g of seeds. Total tocopherol contents ranged from 135 to 525 mg/100 g of oil, with g-tocopherol as the main component, and with different compositional ratios between varieties. Concerning sterols in the oil, total amounts ranged from 364 to 553 mg/100 g, with a predominance of b-sitosterol. After performing principal component analysis, intercultivar differences were found, a potential tool for cultivar authenticity purposes. Moreover, the ingestion of pomegranate arils, with their seeds, increases their beneficial health properties.

In the present work, we investigate how the degree of ripeness (low, low-medium, medium and medium-high) aff ects the physical and compositional changes, as well as antioxidant properties of pomegranate fruit (cv. Mollar de Elche). The skin, pellicle, seed and juice were analysed. The fruit mass increased and the fruit skin became reddish (higher a* and lower h*) as the fruit ripening progressed. The lowest concentrations of fl avonoids and hydrolysable tannins were recorded in skin and pellicles at medium-high maturity stage, which explains the decrease in the total phenols and reducing power during ripening of pomegranate. On the contrary, the highest concentration of fl avonoids (165 mg of quercetin equivalents per 100 mL of juice) was determined in the juice at the most advanced ripening stage, concomitant with the highest total phenols (1695 mg of gallic acid equivalents per 100 mL of juice). Higher DPPH scavenging activity and an increase in the reducing power of juice were also observed during ripening. The trend of the above-mentioned properties allowed describing the fruit development and maturity.

Muitos grupos de insetos necessitam de fontes alimentares que contenham hidratos de carbono, vitaminas e micronutrientes para desempenharem normalmente a suas funçoes biológicas. No ecossistema olival as plantas productoras de pólen podem fornecer esses alimentos. Por outro lado, o pólen constitui um marcador natural, que quando presente no exterior ou no tubodigestivo de diferentes organismos, é frequentemente utilizado em estudos dedispersao da fauna, da sua alimentaçao ou até em estudos peleoecológicos.

This study aims to evaluate the antioxidant potential of extracts of Brassica oleracea L. var. acephala DC. (kale) and several materials of Pieris brassicae L., a common pest of Brassica cultures using a cellular model with hamster lung fibroblast (V79 cells) under quiescent conditions and subjected to H2O2-induced oxidative stress. Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and glutathione was determined by the 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB)-oxidized glutathione (GSSG) reductase recycling assay. The phenolic composition of the extracts was also established by HPLC-DAD. They presented acylated and non acylated flavonoid glycosides, some of them sulfated, and hydroxycinnamic acyl gentiobiosides. All extracts were cytotoxic by themselves at high concentrations and failed to protect V79 cells against H2O2 acute toxicity. No relationship between phenolic composition and cytotoxicity of the extracts was found. Rather, a significant increase in glutathione was observed in cells exposed to kale extract, which contained the highest amount and variety of flavonoids. It can be concluded that although flavonoids-rich extracts have the ability to increase cellular antioxidant defenses, the use of extracts of kale and P. brassicae materials by pharmaceutical or food industries, may constitute an insult to health, especially to debilitated individuals, if high doses are consumed.

Ficus species possess latex-like material within their vasculatures, affording protection and self-healing from physical attacks. In this work, metabolite profiling was performed on Ficus carica latex. Volatiles profile was determined by HS-SPME/GC−IT-MS, with 34 compounds being identified, distributed by distinct chemical classes: 5 aldehydes, 7 alcohols, 1 ketone, 9 monoterpenes, 9 sesquiterpenes and 3 other compounds. Sesquiterpenes constituted the most abundant class in latex (ca. 91% of total identified compounds). Organic acids composition was also characterized, by HPLC−UV, and oxalic, citric, malic, quinic, shikimic and fumaric acids were determined. Malic and shikimic acids were present in higher amounts (ca. 26%, each). The antioxidant potential of this material was checked by distinct in vitro chemical assays. A concentration-dependent activity was noticed against DPPH, nitric oxide and superoxide radicals. Additionally, acetylcholinesterase inhibitory capacity was evaluated, but a weak effect was found.

For the first time, an insect−plant system, Pieris brassicae fed with Brassica rapa var. rapa, was tested for its biological capacity, namely, antioxidant (DPPH•, •NO, and O2•− radicals) and antimicrobial (bacteria and fungi) activities. Samples from the insect’s life cycle (larvae, excrements, exuviae, and butterfly) were always found to be more efficient than the host plant. Also, P. brassicae materials, as well as its host plant, were screened for phenolics and organic acids. The host plant revealed higher amounts of both compounds. Two phenolic acids, ferulic and sinapic, as well as kaempferol 3-O-sophoroside, were common to insect (larvae and excrements) and plant materials, with excrements being considerably richer. Detection of sulfated compounds in excrements, absent in host plant, revealed that metabolic processes in this species involved sulfation. Additionally, deacylation and deglycosilation were observed. All matrices presented the same organic acids qualitative profile, with the exception of excrements.

The antioxidant potential of an aqueous extract obtained from Pieris brassicae larvae reared on Brassica oleracea L. var. costata DC was evaluated against 2,2-diphenyl-1-picrylhydrazyl radical and several reactive oxygen species. The results revealed an effective concentration-dependent protective activity against superoxide and hydroxyl radicals, being superior to that of the host plant. In addition, the larvae extract also exhibited a strong inhibitory effect on xanthine oxidase that was not observed for B. oleracea var. costata. A weak scavenging ability was noticed for hypochlorous acid. Several phenolic compounds with complex chemical structures that are hard to synthesize in the laboratory were found in P. brassicae extract. This is the first time that an insect has been tested for its xanthine oxidase inhibitory capacity, which proved to be very high. These findings are interesting considering that they can be used by food or pharmaceutical industries to prevent the oxidation of their products, to increase the dietary supply of antioxidants, or for prevention of free radical-mediated diseases, namely, gout.

Seeds of Brassica oleracea var. acephala (kale) were analyzed by HPLC/UV-PAD/MSn-ESI. Several phenolic acids and flavonol derivatives were identified. The seeds of this B. oleracea variety exhibited more flavonol derivatives than those of tronchuda cabbage (Brassica oleracea var. costata), also characterized in this paper. Quercetin and isorhamnetin derivatives were found only in kale seeds. Oxalic, aconitic, citric, pyruvic, malic, quinic, shikimic, and fumaric acids were the organic acids present in these matrices, malic acid being predominant in kale and citric acid in tronchuda cabbage seeds. Acetylcholinesterase (AChE) inhibitory activity was determined in aqueous extracts from both seeds. Kale leaves and butterflies, larvae, and excrements of Pieris brassicae reared on kale were also evaluated. Kale seeds were the most effective AChE inhibitor, followed by tronchuda cabbage seeds and kale leaves. With regard to P. brassicae material, excrements exhibited stronger inhibitory capacity. These results may be explained by the presence of sinapine, an analogue of acetylcholine, only in seed materials. A strong concentration-dependent antioxidant capacity against DPPH, nitric oxide, and superoxide radicals was observed for kale seeds.

The phenolic compounds produced by in vitro shoots of Brassica oleracea L. var. costata DC were screened by HPLC-DAD-MS/MS. Thirty seven compounds were characterized, which included chlorogenic acids, flavonoids (the majority of them were hydroxycinnamic acid esters of kaempferol and quercetin glycosides) and hydroxycinnamic acyl glycosides (with predominance of synapoyl gentiobiosides). The antioxidant capacity of the shoots was assessed against DPPH radical and two reactive oxygen species (superoxide radical and hypochlorous acid). A strong concentrationdependent antioxidative capacity was verified in the DPPH and superoxide radicals assays, but a reduced effect was noticed against hypochlorous acid. The results obtained indicate that the in vitro production of B. oleracea var. costata shoots can become important in the obtention of a noticeable dietary source of compounds with health protective potential.

Changes in the phenolics composition of Pieris brassicae larvae fasted for distinct periods (1, 2, 4, 6, and 8 h) and their excrements and of Brassica oleracea L. var. costata DC leaves were determined by high-pressure liquid chromatography/UV-photo diode array detector/mass spectrometry−electrospray ionization. This is the first report following phenolics' metabolism by P. brassicae through time. The results evidence that P. brassicae sequesters and metabolizes the phenolic compounds from the host plant. In a general way, deacylation was the main metabolic reaction that took place, but deglycosylation and sulfate conjugation reactions also occur. Additionally, several kaempferol derivatives containing rhamnose, which is not common in Brassica, were found in the host plant. Attending to the bioactivities recognized for the type of identified compounds, the different materials may constitute an interesting source of bioactive compounds, namely, of highly glycosylated and acylated kaempferol and quercetin derivatives, constituting an economic advantage for producers who have great losses caused by this pest. In addition, a deeper understanding of phenolics metabolism in insects was pursued.

In this work were studied the phenolic composition of in vitro material (shoots, calli, and roots) of Brassica oleracea var. costata and its antioxidant capacity. Samples were obtained in different culture medium, with distinct supplementations to verify their influence on those parameters. Phenolic determination was achieved by HPLC-DAD. Antioxidant activity was assessed against DPPH•. In calli and roots no phenolic compound was identified. In shoots was verified the presence of 36 compounds, which included hydroxycinnamic acids, flavonoids (kaempferol and quercetin derivatives), and hydroxycinnamic acyl glycosides (with a predominance of synapoyl gentiobiosides). MS liquid medium supplemented with 2 mg/L benzylaminopurine (BAP) and 0.1 mg/L naphthaleneacetic acid (NAA) revealed to be the best in vitro condition to produce shoot material with highest phenolic compound contents and stronger antioxidant potential, thus with a possible increase of health benefits.

Phenolic and organic acid profiles of aqueous extracts from Pieris brassicae material and the host kale (Brassica oleracea L. var. acephala) leaves were determined by HPLC/UV–DAD/MSn-ESI and HPLC–UV, respectively. The identified phenolics included acylated and nonacylated flavonoid glycosides, hydroxycinnamic acyl gentiobiosides, and sulphate phenolics. Kale exhibited the highest content (11 g/kg lyophilized extract), while no phenolics were identified in the butterflies or exuviae. Nine different organic acids were characterized in the materials, with kale showing the highest amount (112 g/kg lyophilized extract). With the exception of the exuviae extract, the rest were screened for bioactivity. Using spectrophotometric microassays, all exhibited antiradical capacity against DPPH and NO in a concentration-dependent way, whereas only kale and excrement extracts were active against superoxide. All displayed activity on intestinal smooth muscle, albeit with distinct relaxation–contraction profiles. Larvae and butterfly extracts were more efficacious for intestinal relaxation than was kale extract, whereas excrement extract evoked only contractions, thus evidencing their different compositions. Collectively, these results show that P. brassicae sequesters and metabolizes kale’s phenolic compounds. Moreover, the extract’s bioactivities suggest that they may constitute an interesting source of bioactive compounds whose complex chemical structures preclude either synthesis or isolation.

The phenolic profile of barley (Hordeum vulgare L.) leaves, seeds, awns, and stems, collected in two different locations from Portugal, was determined by a high-performance liquid chromatography/diode array detector (HPLC/DAD). A total of 28 compounds were identified and quantified, which included 4 phenolic acids, 6 C-glycosylflavones, and 18 O-glycosyl-C-glycosyl flavones, with some of them acylated. Distinct profiles were noticed among the analyzed materials. The greatest diversity of compounds was found in barley leaves (26 flavonoids and 2 phenolic acid derivatives), which also exhibited the highest concentration of phenolics. Isoorientin-7-O-glucoside (lutonarin) was the major compound in leaves, while, in general, the pair isovitexin-7-O-rutinoside plus isoscoparin-7-O-glucoside were the main phenolics in the other materials. Thus, barley leaves may constitute an important dietary source of protective compounds, which could be used, for example, to take profit from the wastes resulting from alcoholic drink obtainment.

The genus Brassica is related with the prevention of carcinomas, especially of stomach, colon and recto and with the prevention of cardiovascular diseases. The most consumed Brassica species in Portugal is the tronchuda cabbage or Portuguese cabbage (Brassica oleracea var. costata DC). This study concerned the characterization of chemical composition (phenolics and organic acids) and the evaluation of the antioxidant potential of tronchuda cabbage. Seeds, sprouts, internal and external leaves were analysed. To evaluate the antioxidant potential, the ability of tronchuda cabbage materials to act as a scavenger of reactive oxygen species (superoxide radical, hydroxyl radical and hypochlorous acid) was investigated. Additionally, it was found that Pieris brassicae, a plague infesting Brassica cultures, exhibits interesting antioxidant potential, once it sequesters, metabolizes and accumulates phenolics. The possibility of commercialization of standardized aqueous extracts of Brassica oleracea var. costata DC seeds, sprouts, internal and external leaves, with nutritional and healthy potential benefits, is discussed.

Two Brassica oleracea varieties (B. oleracea L. var. costata DC and B. oleracea L. var. acephala) and Brassica rapa L. var. rapa inflorescences were studied for their chemical composition and antioxidant capacity. Phenolic compounds and organic acids profiles were determined by HPLC–DAD and HPLC–UV, respectively. B. oleracea var. costata and B. oleracea L. var. acephala inflorescences presented a similar qualitative phenolic composition, exhibiting several complex kaempferol derivatives and 3-p-coumaroylquinic acid, while B. rapa var. rapa was characterized by kaempferol and isorhamnetin glycosides and several phenolic acids derivatives. B. oleracea L. var. costata and B. rapa var. rapa showed the highest phenolics content. The three Brassica exhibited the same six organic acids (aconitic, citric, pyruvic, malic, shikimic and fumaric acids), but B. oleracea L. var. acephala presented a considerably higher amount. Each inflorescence was investigated for its capacity to act as a scavenger of DPPH radical and reactive oxygen species (superoxide radical, hydroxyl radical and hypochlorous acid), exhibiting antioxidant capacity in a concentration dependent manner against all radicals. These samples were also studied for its antimicrobial potential against Gram-positive and Gram-negative bacteria and fungi, displaying antimicrobial capacity only against Gram-positive bacteria.

The phenolic profiles of cabbage white butterfly (Pieris brassicae L.; Lepidoptera: Pieridae) at different development stages (larvae, exuviae, and butterfly), its excrements, and its host plant Brassica rapa var. rapa L. were determined by high performance liquid chromatography− diode-array detector−mass spectrometry/mass spectrometry−electrospray ionization (HPLC-DAD-MS/MS-ESI). Twenty-five acylated and nonacylated flavonoid glycosides and ferulic and sinapic acids were identified in host plant, from which only 12 compounds were found in the excrements. In addition, the excrements showed the presence of sulfate flavonoids and other flavonoid glycosides that were not detected in the leaves. In the larvae kept without food for 12 h, only 3 compounds common to the plant material and 2 others, also present in the excrements, were characterized. The results indicate that deacylation, deglycosylation, and sulfating steps are involved in the metabolic process of P. brassicae and that its excrements may constitute a promising source of bioactive compounds, which could be used to take profit of this common pest of Brassica cultures.

A field experiment was carried out to investigate the effect of fertilization level on the phenolic composition of tronchuda cabbage (Brassica oleracea L. var. costata DC) external and internal leaves. Eight different plots were constituted: a control without fertilization, one with organic matter, and six experiments with conventional fertilizers (nitrogen, boron, and sulfur, two levels each). The phenolic compounds were analyzed by reversed-phase HPLC-DAD. External and internal leaves revealed distinct qualitative composition. In the internal leaves were found 15 phenolics (5 kaempferol and 10 cinnamic acid derivatives), whereas the external leaves presented 3-p-coumaroylquinic acid and 13 kaempferol derivatives. Principal component analysis (PCA) was applied to assess the relationships between phenolic compounds, agronomical practices, and harvesting time. Samples obtained with conventional practices were quite effectively separated from organic samples, for both types of leaves. In general, samples developed without any fertilization presented the highest phenolics amounts: external and internal leaves contained 1.4- and 4.6-fold more phenolic compounds than the ones that received conventional fertilizer, respectively, and the internal leaves presented 2.4 times more phenolics than the ones grown with organic amendment. Additionally, samples from organic production exhibited higher total phenolics content than those from conventional practices, collected at the same time. Samples harvested first were revealed to be distinct from the ones collected later. The results show that it is possible to grow tronchuda cabbage without excess fertilizers, with highest amounts of phenolics and reduced environment contamination.

The changes in antioxidant compounds of Brassica oleracea L. var. costata DC seeds were monitored during the first twelve days of seedling development. Sprouts were screened at time intervals of two days for phenolic compounds and organic acids. The identified phenolic compounds included esters of sinapic acid with glucose, gentiobiose and kaempferol, as well as sinapoylcholine. The organic acids were oxalic, aconitic, citric, pyruvic, malic, shikimic, and fumaric acids. During germination, a depletion of phenolic compounds was observed, although no qualitative changes were seen. Among individual compounds, kaempferol, choline and glucose esters of sinapic acid showed a marked decrease between days two and six, whereas the changes in gentiobiose esters of sinapic acid were smaller. The total organic acids content increased rapidly during the first four days, with less significant variations thereafter. Malic acid, the major organic acid found in sprouts, greatly contributed to this result though oxalic, pyruvic, and fumaric acids also increased in the same manner. In contrast, aconitic, citric and shikimic acids showed decreases between days two and twelve of germination.

The flavonoid pattern of larvae of cabbage white butterfly (Pieris brassicae L.; Lepidoptera: Pieridae) reared on the leaves of tronchuda cabbage was analysed by HPLC-DAD-MS/MS-ESI. Twenty flavonoids were identified or characterised, namely 16 kaempferol and four quercetin derivatives. Kaempferol 3-O-sophoroside, a minor component of tronchuda cabbage, was found to be the main component in P. brassicae (15.8%). Apart from this, only two other flavonoids present in significant amounts in tronchuda cabbage (kaempferol 3-O-sophoroside-7-O-glucoside and kaempferol 3-O-sophoroside-7-O-sophoroside) were found in the larvae. The larvae have high amounts of quercetin derivatives (18.5%), which were present only in trace amounts in tronchuda cabbage extracts, suggesting that P. brassicae is able to selectively sequester these flavonoids. The occurrence of a high content of flavonoids not detectable in tronchuda cabbage extracts indicates that P. brassicae larvae are able to metabolize dietary flavonoids., [Graphical abstract]: The flavonoid pattern of larvae of cabbage white butterfly (Pieris brassicae L.; Lepidoptera: Pieridae) reared on the leaves of tronchuda cabbage was analysed by HPLC-DAD-MS/MS-ESI. Twenty kaempferol and quercetin derivatives were identified or characterised. The results suggest that P. brassicae is able to selectively sequester quercetin derivatives and to metabolize dietary flavonoids.