Antioxidant,antimicrobial and toxicological properties of Schinus molle L. essential oils

Ethnopharmacological relevance: Schinus molle L. has been used in folk medicine as antibacterial, antiviral, topical antiseptic, antifungal, antioxidant, anti-inflammatory, anti-tumoural as well as antispasmodic and analgesic; however, there are few studies of pharmacological and toxicological properties of S. molle essential oils. Aim of the study: The aim of this study was to evaluate the antioxidant and antimicrobial activities of S. molle leaf and fruit essential oils, correlated with their chemical composition and evaluate their acute toxicity. Materials and methods: The chemical composition of S. molle leaf and fruit essential oils were evaluated by GC-FID and GC-MS. Antioxidant properties were determined using the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) free radical and β-carotene/linoleic acid methods. Antimicrobial properties were evaluated by the agar disc diffusion method and minimal inhibitory concentration assay. Toxicity in Artemia salina and acute toxicity with behavioural screening in mice were evaluated. Results: The dominant compounds found in leaf and fruit essential oils (EOs) were monoterpene hydrocarbons, namely -phellandrene, β-phellandrene, β-myrcene, limonene and α-pinene. EOs showed low scavenging antioxidant activity by the DPPH free radical method and a higher activity by the β-carotene/linoleic acid method. Antimicrobial activity of EOs was observed for Gram+, Gram– pathogenic bacteria and food spoilage fungi. EOs showed cytotoxicity for Artemia salina and lower toxicity in Swiss mice. Conclusions: The result showed that EOs of leaves and fruits of S. molle demonstrated antioxidant and antimicrobial properties, suggesting their potential use in food or pharmaceutical industries.

EFFECT OF DIFFERENT STORAGE CONDITIONS ON NUTRITIONAL AND QUALITY PARAMETERS OF 'SWEETHEART' CHERRY.

Abstract The sweet cherry ‘Sweetheart’, although having a short shelf life, is highly appreciated by consumers due to its organoleptic characteristics. Different storage methods were tested to study the maintenance of quality during a period of 27 days: 1) cold (air at 1°C and 95% relative humidity) (CC), 2) cold and polypropylene film bags (1°C and 95% relative humidity) (MA) and 3) cold and controlled atmosphere (1°C, 95% RH, 10% CO2 and 8% O2) (CA). Quality parameters tested included external colour (L*, a*, b*), total soluble solids (TSS), and titratable acidity (TA). To evaluate nutritional quality anthocyanins, total antioxidant activity, and total phenolics were measured. Results allow us to say that phenolic compounds were relatively stable and similar during storage in CC and MA. Cherries stored under CA conditions presented lowest concentrations of phenolic compounds. Phenolic compounds, total anthocyanins and antioxidant activity were inversely correlated with values of colour coordinates. Considering all the evaluations done during this work it is unquestionable that fruits stored in controlled atmosphere conditions had significantly different quality.

ANTIOXIDANT, ANTIMICROBIAL AND TOXICOLOGICAL PROPERTIES OF SCHINUS MOLLE L. ESSENTIAL OILS

Schinus molle L. is commonly known as pink pepper or American pepper, of Anacardiaceae family, from subtropical regions of South America, introduced and naturalized in South Europe, including Portugal. In folk medicine, plant extracts and essential oil has related as having antibacterial, antiviral, antifungal, anti-inflammatory, antitumoral, antispasmodic, analgesic and antidepressive properties. The aim of present study was to evaluate the chemical composition and biological activities of essential oil extracted from leaves and fruits of S. molle. For this purpose, the essential oils were analyzed by gas chromatography (GC/FID) and antioxidant properties were evaluated by the free radical DPPH and by system β-carotene/linoleic acid methods. The antimicrobial activities were screened against pathogenic bacteria and fungi and food spoiling fungi by the disc diffusion assay and minimal inhibitory concentration (MIC) was determined for sensitive strains. Toxicity of essential oils were carried out by the brine shrimp mortality test (EC50) and acute lethal dose (DL50) determination after oral administration in Swiss mice The major components in leaf essential oil were α-phellandrene, β-phellandrene and limonene, while myrcene, α-phellandrene and 1,8-cineole are the main components in the fruit essential oil. The essential oils of leaf and fruit of S. molle showed antioxidant activity through the two mechanisms: the ability to capture free radicals and protection of lipid peroxidation. These oils exhibited also a broad microbial activity spectrum, against pathogenic bacteria Gram-positive and Gram-negative and Candida spp. The fruit essential oil showed high cytotoxicity against Artemia salina. Essential oils of leaves and fruits of S. molle showed significant antioxidant and microbial properties, so the studies continue to clarify more in deep its toxicity, including hepatotoxicity and nephrotoxicity, and to evaluate its medicinal or nutraceutical potential.

Evaluation of different postharvest conditions to preserve the amount of bioactive compounds, physicochemical quality parameters and sensory attributes of ‘Sweetheart’ cherries.

Sweet cherries (Prunus avium L.) ‘Sweetheart’ were harvested at different production regions from Portugal (Cova da Beira and Portalegre) and Spain (Valle de Jerte). Cherries were harvested at their commercial maturation according to the empirical knowledge of external color corresponding to good quality. Fruits were stored and evaluated in order to study their quality on the harvest day and during a period of 21 days, at cold storage (1 ºC, 95% RH). The sweet cherry ‘Sweetheart’ is a well-known variety and a highly appreciated one but fruits present a short shelf life. On the other hand the effect of different “terroir” on cherry characteristics should be known and clarified. Fruits from day 0, considered without storage, were kept at 20ºC and analyzed. Every weak, 3 replicas were randomly picked up and 10 fruits from each one were submitted to several analyses after fruit temperature stabilized at 20ºC. Several quality parameters were evaluated: external colour (L*, a*, b*), texture, soluble solids content (SSC), titratable acidity (TA) and the ratio between soluble solid contents (SSC) and tritratable acidity (TA). Fruits from different orchards and locations were significantly different according to these parameters. Fruits from Cova da Beira were less firm comparing with other two regions, Valle de Jerte and Portalegre, which may indicate a higher maturation rate at harvest in those fruits. This is in accordance with SSC/titratable acidity rate suggesting a late harvest in Cova da Beira comparing with other two orchards, however fruits from Cova da Beira exhibit a poor color at harvest. These results clearly showed a lower correlation between SSC and firmness considering fruits origin.

Quality of cherry ‘Sweetheart’ from different regions of Portugal and Spain (Cova da Beira, Portalegre and Valle del Jerte).

Sweet cherries (Prunus avium L.) ‘Sweetheart’ were harvested at different production regions from Portugal (Cova da Beira and Portalegre) and Spain (Valle de Jerte). Cherries were harvested at their commercial maturation according to the empirical knowledge of external color corresponding to good quality. Fruits were stored and evaluated in order to study their quality on the harvest day and during a period of 21 days, at cold storage (1 ºC, 95% RH). The sweet cherry ‘Sweetheart’ is a well known variety and a highly appreciated one but fruits present a short shelf life. On the other hand the effect of different “terroir” on cherry characteristics should be known and clarified. Fruits from day 0, considered without storage, were kept at 20ºC and analyzed. Every weak, 3 replicas were randomly picked up and 10 fruits from each one were submitted to several analyses after fruit temperature stabilized at 20ºC. Several quality parameters were evaluated: external color (L*, a*, b*), texture, soluble solids content (SSC), titratable acidity (TA) and the ratio between soluble solid contents (SSC) and tritratable acidity (TA). Fruits from different orchards and locations were significantly different according to these parameters. Fruits from Cova da Beira were less firm comparing with other two regions, Valle de Jerte and Portalegre, which may indicate a higher maturation rate at harvest in those fruits. This is in accordance with SSC/titratable acidity rate suggesting a late harvest in Cova da Beira comparing with other two orchards, however fruits from Cova da Beira exhibit a poor color at harvest. These results clearly showed a lower correlation between SSC and firmness considering fruits origin.

QUALITY OF 'SWEETHEART' CHERRY UNDER DIFFERENT STORAGE CONDITIONS

Abstract: In Portalegre, Portugal, sweet cherry production is very important to the region’s economic sustainability. The sweet cherry ‘Sweetheart’ has exhibited short shelf life in spite of being highly appreciated by consumers due to its organoleptic characteristics. In this trial, we evaluated fruit quality of ‘Sweetheart’ stored under different storage conditions: 1) cold conditions (1ºC and high humidity 95%), 2) cold conditions and polypropylene film bags (MA), and 3) controlled atmosphere (CA) (1°C, 95% humidity, 10% CO2 and 8% O2). Fruit physical and chemical parameters were evaluated after 0, 6, 13, 20 and 27 days of cold storage. Quality parameters tested included weight loss, external colour (L* a* b*), visual assessment of the epidermis, epidermis and mesocarp penetration test, soluble solids content (SSC), and titratable acidity (TA). We also performed sensory analyses. The results for textural properties, colour coordinates and sensory analysis suggest that ‘Sweetheart’ fruit can be stored under cold conditions, 1°C, 95% humidity, for up to 21 days without significant loss of quality. Controlled atmosphere maintained tissue turgidity during storage; however, this was not noticed by the panelists, who consistently classified fruits stored under CA conditions with lower overall ratings than fruits under cold conditions with or without film bags.

Comportamento pós-colheita da ameixa 'Raínha Cláudia Verde' Prunus domestica L. : efeitos do cálcio na maturação

`Rainha Cláudia Verde' é uma antiga variedade da ameixeira europeia Prunus domestica L. que se encontra bem adaptada a uma zona restrita do Alto Alentejo. Esta variedade é utilizada para o consumo em fresco e na doçaria regional, onde se emprega na confecção das famosas 'Ameixas D'Elvas'. A confitagem é a técnica utilizada na confecção deste produto com denomicação de origem protegida, e que faz parte de um saber tradicional muito divulgado na região. As informações que resultam de evidências práticas mostram que nem todos os frutos reagem da mesma forma à fase da cozedura. Existem zonas específicas que produzem frutos que não se adequara processo da confitagem, apresentando uma textura imprópria após a cozedura. Recentemente, e com o aumento das áreas produtoras, verificou-se que os frutos destas regiões específicas para além de inadequados para a cozedura, também apresentavam uma menor capacidade de conservação em fresco. Assim durante a conservação estes frutos, quando comparados com os das outras regiões, apresentavam uma perda de firmeza mais rápida tornando-se mais difíceis de comercializar. Entre os factores culturais que contribuem para a qualidade pós-colheita dos frutos, o teor de cálcio presente no solo e nos frutos apresenta-se corno um dos mais importantes. O cálcio é um dos nutrientes que mais frequentemente é associado à manutenção da estrutura das paredes celulares das plantas, estando envolvido directamente na redução das perdas de textura dos frutos. Tendo em consideração os aspectos anteriormente referidos, foi delineado um trabalho que teve início com a selecção de dois pomares geograficamente distantes, e que tradicionalmente produziam frutos com diferentes comportamentos quer durante a conservação quer durante a confitagem. Associada ao conhecimento empírico, a prévia indicação de que estes pomares apresentavam concentrações cle cálcio foliar significativamente diferentes, contribuiu também para a sua selecção. O objectivo geral desta tese foi o de investigar o comportamento pós-colheita da `Rainha Cláudia Verde' particularmente a influência do cálcio na textura dos frutos. Definiram-se os seguintes objectivos específicos: (1) determinar a influência do porta-enxerto e do solo na concentração de cálcio dos frutos e as respectivas consequências, no seu comportamento pós-colheita; (2) seleccionar um método que permitisse avaliar a produção de etileno dos frutos e consequentemente a atribuição da designação de fruto climatérico ou não climatérico a esta variedade; (3) avaliar os e Feitos durante a conservação, de diferentes níveis de cálcio nos frutos; (4) quantificar nos frutos os níveis de cálcio da parede celular e avaliar a sua influência na firmeza dos frutos; (5) seleccionar as melhores temperaturas de conservação para os frutos desta variedade. Os resultados apresentados nesta tese indicam que a variedade 'Rainha Cláudia Verde' é uma variedade de frutos climatéricos que apresentam uma acentuada perda de textura após a colheita. Durante a conservação frigorífica os frutos apresentaram comportamentos diferentes, de acordo com a sua origem. Os frutos com origem no pomar que tradicionalmente não produz frutos aptos a serem confitados, apresentam simultaneamente uma mais rápida perda de firmeza quando comparados com os frutos dos outros pomares. Sendo que o menor teor de cálcio nos frutos leva a que a diminuição da firmeza da polpa ocorra mais rapidamente. No entanto à colheita não se observaram diferenças significativas da firmeza da polpa dos frutos. Esta informação parece indicar que outros factores, além do nível de cálcio dos frutos, poderão estar implicados na firmeza revelada à colheita. Por outro lado os frutos com epiderme revelaram diferenças de firmeza nos testes efectuados à colheita. O efeito da epiderme na firmeza dos frutos à colheita parece indicar que outros factores tais como o estado de hidratação dos frutos poderão contribuir para o aumento desta característica dos frutos. Apesar do teor de cálcio dos frutos melhorar o seu comportamento durante a conservação, a sua influência na emissão de etileno não foi evidente. O aumento do teor de cálcio nos frutos pode conseguir-se através de uma selecção adequada do porta-enxerto. Os porta-enxertos estudados induziram quantidades diferentes de cálcio nos frutos, aparentemente contribuindo o vigor do porta-enxerto para um efeito de diluição do cálcio na árvore. O estudo da influência do solo no teor do cálcio dos frutos revelou que os frutos com menor capacidade de conservação provinham de solos com maiores teores de cálcio, e que na sua constituição apresentavam uma menor concentração de cálcio na polpa. Apresenta-se ainda a hipótese de que o excesso de potássio presente nestes solos possa ter contribuído para um menor teor de cálcio nos frutos. De facto os frutos com uma razão Ca/K superior apresentaram também uma firmeza superior./ ABSTRACT - `Raínha Claudia Verde' is an old variety of Prunus domestica which is well adapted to a restrict zone of Alto Alentejo in the south region of Portugal. This variety is much appreciated either as a fresh fruit or as a sweet candy. The candying process is a widespread technique in this region with much empirical knowledge. There are practical evidences which indicate that fruits origin may influence the boiling process. Some fruits produced in specific areas in this region had an inadequate behaviour during boiling, becoming to soft and improper to use in canding. More recently it has been also observed that these specific areas produced fruits with a poor postharvest behaviour. During storage these fruits loose texture very quickly and became improper to commercialize. Many pre and postharvest factors may contribute to differences in fruit quality. calcium is one of most important nutrients which have a major effect on cell wall structure and membrane integrity. Studies on the role of calcium in fruits indicate its involvement in delaying changes associated with softening. Two orchards were selected because of their history of producing fruits with different characteristics either as a fresh or as a processed fruit, and because induced different calcium levels in the leaves. The main focus of this research work was to study the influence of the production region in fruit postharvest behaviour, specially the influence of calcium in fruit texture. The aims were: (1) to compare the rootstock and the soil influence on calcium fruit content, (2) to select a method- to measure the production of ethylene in fruits of 'Raínha Claudia Verde' (3) to evaluate the effects of different calcium fruit content in the postharvest behaviour of fruits (4) to evaluate the cell wall calcium content and its influence in fruit firmness, (5) and to select the best cold storage temperatures to this variety. It was found that 'Raínha Claudia Verde' is a climacteric variety and the studies on fruit firmness revealed a significant loss of fruit texture during ripening on or off the tree. During storage, fruits had a different behaviour depending on fruits origin. Usually fruits, which traditionally do not resist to boiling process, also exhibited an early softening, when compared to other fruits produced in adequate regions. The excessive fruit softening after harvest occurred in fruits with lower calcium content. However, at harvest, fruits from both orchards exhibited a similar firmness which may indicate that other factors besides calcium should be implicated in fruit firmness at harvest. In spite of a better postharvest behaviour of fruits with higher calcium content, it was not evident the calcium influence in the climacteric rise. The increase of calcium fruit content can be achieved with a proper rootstock selection. The rootstocks investigated in this study, induced different calcium fruit content, apparently vigorous rootstocks contributed to the dilution of calcium fruit level. The soil with higher calcium content induced a lower calcium fruit content, which may be due to the excess of potassium in this soil; in fact fruits with higher Ca/K ratio reached higher firmness values. It is also proposed a method to evaluate the calcium content in the fruit independently of fruit mass. The calcium fruit content is usually expressed as a percentage of dry mass, however during the course of fruit development there are a huge increment of fruit weight because of water and sugar mobilization into the fruit. Most of the total calcium in the plants is associated with the cell wall which means that calcium fruit content expressed as a percentage of cell wall fraction is a much more reliable method. Orchards with an excess of potassium in the soil produced fruits with a significant lower calcium fruit content. However it was not possible to prove the gradually firmness decrease during the harvest period as a consequence of calcium fruit loss. In fact, it was not evident a gradual decrease of calcium fruit content during the harvest dates thus it was impossible to find at harvest, a good correlation between fruit firmness and calcium fruit content. The analysis of cell wall polysaccharides evaluated during ripening in the tree showed a slight increase of more branched polysaccharides as ripening went. The small changes in pectic polysaccharides during the harvest season are in accordance with the small decrease in tissue firmness during this period. In this variety the usual storage period is about of 3-4weeks with a temperature of 1-2°C and 90% of relative humidity. However upon rewarming fruits held at 7°C, during 14 days, produced more ethylene at 20°C and exhibited also a higher firmness than fruits held at PC. The reduction of ethylene production and fruit firmness upon rewarming, after fruits being held at lower temperatures, may suggest some chilling injury in this variety.

Consumers sensory evaluation of melon sweetness and quality

CONSUMERS SENSORY EVALUATION OF MELON SWEETNESS AND QUALITY Agulheiro Santos, A.C, Rato, A.E., Laranjo, M. and Gonçalves, C. Departamento de Fitotecnia, Escola de Ciências e Tecnologia, Instituto de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM), Instituto de Investigação e Formação Avançada (IIFA), Universidade de Évora, Polo da Mitra, Ap.94, 7002-554 Évora, Portugal. ABSTRACT The sensory quality of fruits is made of a range of attributes like sweetness, acidity, aroma, firmness, color. Taste perception and perception threshold of these attributes are variable according to the psychological and cultural development of individuals. To better understand the quality evaluation of melon by consumers, consumers were invited to taste melon samples, in supermarkets in Évora (South region), Lisbon (Central region) and Vila Nova de Gaia (North region). The present work explored the importance given by consumers to sweetness in order to classify the overall quality of melon. Furthermore, the relationship of the chemical evaluation of Total Soluble Solids (TSS) with sweetness of melon was studied. Fruits from the variety Melão branco picked randomly from those that were exposed for sale in supermarkets were used for analysis. Fruits were chinned along the equatorial zone and only the central part of the fruit, opposite to the part that leaned on the soil, was used to obtain homogeneous samples. Consumers were invited to taste four small pieces of each fruit, previously referenced with a code number, and answer a questionnaire with two questions related to sweetness and overall quality. Each question had five possible levels, identified from “Nothing sweet”, to “Extremely sweet”, in one case, and from “Poor” to “Excellent” in the other. Simultaneously, the values of TSS (measured in ºBrix) for each melon used in the study were evaluated by refractometry. This sensory analysis allowed us to point out the following findings: first of all, there is good agreement between the results obtained to classify “Sweetness” and “Overall Quality” (Cohen’s Kappa=53.1%, p<0.001), which means, for example, that fruits with excellent quality are in general extremely sweet. Moreover, fruits with less than 9.6 °Brix are considered of poor quality and nothing sweet, whereas fruits with values between 10 °Brix and 12 °Brix are considered good in terms of overall quality. It seems that the thresholds for the stimulus/intensity of sweetness lied between 10 °Brix to 14 °Brix for this melon variety. Acknowledgments This work was support by national funds through Fundação para a Ciência e a Tecnologia (FCT) under the Strategic Project Pest-OE/AGR/UI0115/2014 and co-funded by FEDER funds through the COMPETE Program.

Effect of storage on physical-chemical properties and phenolics of sweet cherry from São Julião region.

Effect of storage on physical-chemical properties and phenolics of sweet cherry from São Julião region. A.C. Agulheiro-Santos1, F. Vieira1, D. Gonzalez2, M. Lozano2, V. Palma1, A.E. Rato1 1Universidade de Évora. Instituto de Ciências Agrárias e Ambientais Mediterrânica (ICAAM). 7000 Évora, Portugal. 2Centro de Investigaciones Científicas y Tecnológicas de Extremadura (CICYTEX). Ctra. San Vicente. Finca Santa Engracia. 06071 Badajoz, Spain. Worldwide the consumption of fruit and vegetables is increasing due to the dietary guidelines recommended by nutritionist. Because of their high content on phenols, vitamins, mineral and antioxidants, berry fruits are consumed not only in fresh forms but also as processed and derivative products such as juices, yogurts, jellies and dried fruits. As a high consumed red fruit, sweet cherry has been the focus on some studies, mainly regarding bioactive compounds content. “Sweetheart” cherries from São Julião region (Alentejo, Portugal) from two different production campaigns were kept in different storage conditions in order to evaluate both the environmental and storage effect on some physical-chemical properties and phenolics. Cold conditions - Cold (1 ºC, 95% RH) and modified atmosphere - MA (1 ºC, 95% RH with micro-perforated bags of Pplus®, Sidlaw Packaging, Bristol, UK) were tested. In order to establish the appropriate storage conditions, individual phenolic acids and physical-chemical properties were analysed during two consecutive years. Results show a general decrease on phenolic compounds content between cherries from both years. It is also observed that MA conditions do not affect significantly both phenolics and physical-chemical parameters when compared with Cold conditions. Additionally, it is observed similar behaviour on Cold and MA sweet cherries regarding its pH, total soluble solids content (TSS), titratable acidity (TA) and colour and individual phenols during storage time. Concluding, these results show, as expected, changes between cultivars which may be correlated with the environmental conditions on different years. Keywords: sweet cherry, postharvest, phenols, physical-chemical, storage conditions.