Implication of adipocytes from subcutaneous adipose tissue and fatty acids in skin inflammation caused by λ-carrageenin in gilthead seabream (Sparusaurata).

The role of subcutaneous adipose tissue adipocytes and the effects of fatty acids on carrageenan-induced skin inflammation in gilthead seabream (Sparus aurata) were studied. Fish were injected intramuscularly with phosphate-buffered saline (control) or λ-carrageenin (1%), and skin samples collected at the injection site at 3 and 6 h post-injection (p.i.) were processed for histological study. In addition, the presence and levels of lipid classes, fatty acid methyl esters (FAME) and eicosanoids were evaluated in the skin samples obtained from the injected areas. Histological results indicated an increase in adipocyte area in fish sampled at 3 h p.i. with λ-carrageenin compared to fish in the control group. Furthermore, the frequency of adipocytes between 4500 and 5000 µm2 was increased at 6 h in the λ-carrageenin group compared to the control group. Analysis of lipid classes found that fish injected with λ-carrageenan showed increased free fatty acid (FFA) and sphingomyelin content at 3 and 6 h, respectively, compared to the control group. An increase in saturated fatty acids (SFA), n-6 polyunsaturated fatty acids (PUFA), and a decrease in the values of monounsaturated fatty acids (MUFA), n-3 PUFA and minor fatty acids were observed in fish skin at 6 h after λ-carrageenin injection, with respect to the values obtained in the control group. Regarding the analysis of eicosanoids, an increase in hydroxyeicosatetraenoic acid (5-HETE) was detected in the skin of fish at 6 h post-carrageenin injection compared to the control group. The presented results indicate the contribution of adipocytes and fatty acids in the development and regulation of the inflammatory response triggered by λ-carrageenin in gilthead seabream skin.

The Green Microalga Coelastrella thermophila var. globulina (Scenedesmaceae, Chlorophyta) Isolated from an Algerian Hot Spring as a Potential Source of Fatty Acids.

Screening microalgae from extreme environments, including hot springs, is an important research topic that has lately emerged. A thermophilic green alga was isolated from a north-eastern Algerian hot spring at a temperature of 63 °C, and its fatty acid (FA) profile was explored. The strain was cultivated in BBM medium at 35 °C in a 16:8 h light/dark cycle and 75 µM photons m-2 s-1. The morphological studies combined with phylogenetic analysis revealed that the isolate was Coelastrella thermophila var. globulina Q. Wang, H. Song, X. Liu, G. Liu and Z. Hu. The monounsaturated fatty acid (MUFA) content was 51.12%. The saturated fatty acid (SFA) and polyunsaturated fatty acid (PUFA) content, respectively, accounted for 27.01% and 21.87%. The main FA was oleic acid (18:1n-9), whose value was 35.95%, followed in decreasing order by palmitic acid (16:0) with 21.45%, linoleic acid (18:2n-6) with 14.38% and α-linolenic acid (18:3n-3) with 04.22%. The FA profile exhibited high total n-6 and n-3 PUFA values (15.80% and 5.76%, respectively). Coelastrella thermophila var. globulina is particularly interesting for producing n-6 and n-3 PUFA and is likely suitable for other biotechnological purposes. This is the first time that this taxon has been reported in hot springs. Other species can be expected to be reported, which emphasises the importance of the biodiversity of extreme habitats.

Acute inflammatory response in the skin of gilthead seabream (Sparus aurata) caused by carrageenin.

Although inflammation is a well-characterized process in mammals, few studies have dealt with the mechanisms involved in this process in fish. The present study evaluated the expression of inflammation-related genes in the skin of fish injected with carrageenin, which has previously been used in inflammatory models in mammals. In our case, fish were injected subcutaneously with PBS (as control) or carrageenin (1%), and skin samples from the injection site were collected 1.5, 3 and 6 h post-injection. The gene expression of inflammatory markers (csfr1, mhc-ii and phox40), several pro-inflammatory cytokines (il1b, tnfa, il6, il8 and il18) and other molecules related (such as myd88 and c-rel) were up-regulated at 1.5 and 3 h in fish injected with carrageenin compared with control levels. By contrast, the gene expression of anti-inflammatory molecules (nlrx1, nlrc5 isoform 1, ctsd and ctss) was down-regulated in fish injected with carrageenin and sampled 3 h post injection, again compared to the gene expression in control fish. According to our results, carrageenin can be considered not only a good stimulator to study skin inflammation in gilthead seabream but also this method might be use to study the modulation of fish inflammatory process caused by internal or external factors.

Immune defence mechanisms presented in liver homogenates and bile of gilthead seabream (Sparus aurata).

Because the role of the liver of fishes in providing possible immunity remains largely unknown, the aim of this work was to identify and characterize different humoral defence mechanisms in the liver homogenates and bile of gilthead seabream (Sparus aurata) for the first time. Total protein levels and several immune parameters (complement activity, lysozyme and immunoglobulin M level) were studied. Furthermore, the activity of some lytic (proteases, antiproteases, esterase, alkaline phosphatase) and antioxidant (superoxide dismutase, catalase and peroxidase) enzymes was determined. Finally, bacteriostatic activity on three opportunist fish pathogens (Vibrio harveyi, Vibrio angillarum and Photobacterium damselae) was measured. Lysozyme and antiprotease activity were undetected in liver and bile, while natural haemolytic complement activity was only detected in bile, and immunoglobulin M was detected in both samples. The levels of proteases, esterase and antioxidant enzymes were greater in bile than in liver homogenates, while the level of alkaline phosphatase was very low in both samples. In addition, while no bacteriostatic activity was detected on liver homogenates, the bile revealed a very potent bacteriostatic activity against all the tested pathogenic bacteria. These results corroborate that fish liver - especially fish bile - contains many factors involved in innate immunity that could be useful for better understanding the role of the liver as an organ involved in fish immune functions as well as the possible contribution of bile to gut mucosal immunity.

Probiotic properties and fatty acid composition of the yeast Kluyveromyces lactis M3. In vivo immunomodulatory activities in gilthead seabream (Sparus aurata).

The aim of this study was to analyze the probiotic potential, fatty acid composition and immunostimulant activities of Kluyveromyces lactis M3 isolated from a hypersaline sediment. For this purpose, K. lactis M3 resistance to different pH, salinities and bile, as well as its antioxidant capability were assayed. Furthermore, total fatty acid composition of the yeast was determined where the dominant fatty acids were palmitic, palmitoleic, oleic and linoleic acids. K. lactis M3 showed no cytotoxic effects on peripheral blood leukocytes. During an in vivo experiment in gilthead seabream (Sparus aurata), dietary K. lactis M3 supplemented at 0.55 or 1.1% of the basal diet enhanced bactericidal activity against Vibrio parahaemolyticus N16, V. harveyi Lg 16/00, and V. anguillarum CECT 43442 compared to fish fed commercial diet (control group). Finally, nitric oxide production, peroxidase activity and skin mucus lectin union levels strongly increased in fish fed K. lactis M3 with respect to the control group. The results suggested that the yeast K. lactis M3 had exhibited high antioxidant capability, and its dietary administration at 0.55 or 1% basal diet had immunostimulant activity for gilthead seabream. For all these reasons, it should be considered an appropriate probiotic candidate for the aquaculture fish industry.

Recombinant nodavirus vaccine produced in bacteria and administered without purification elicits humoral immunity and protects European sea bass against infection.

Viral necrosis virus (NNV) or nodavirus causes fish viral encephalopathy and retinopathy worldwide. In some cases, mortalities in aquaculture industry can reach up to 100%, some species being especially sensitive as is the case of European sea bass (Dicentrarchus labrax), one of the main cultured species in the Mediterranean, with the consequent economical loses. Development of new vaccines against NNV is in the spotlight though few researches have focused in European sea bass. In this study we have generated a recombinant NNV (rNNV) vaccine produced in Escherichia coli expressing the capsid protein and administered it to European sea bass juveniles by two different routes (intraperitoneal and oral). The last being considered non-stressful and desired for fish farming of small fish, which in fact are the most affected by NNV. Oral vaccine was composed of feed pellets containing the recombinant whole bacteria, and injected vaccine was composed of recombinant bacteria previously lysed. Our results revealed production of specific anti-NNV IgM following the two vaccination procedures, levels that were further increased in orally-vaccinated group after challenge with NNV. Genes related to interferon (IFN), T-cell and immunoglobulin markers were scarcely regulated in head-kidney (HK), gut or brain. Vaccination by either route elicited a relative survival response of 100% after NNV challenge. To our knowledge, this is the first report of a recombinant vaccine followed by no purification steps which resulted in a complete protection in European sea bass when challenged with NNV.

Age-related changes in mitochondrial membrane composition of Nothobranchius furzeri.: comparison with a longer-living Nothobranchius species.

Membrane compositions, particularly of mitochondria, could be critical factors in the mechanisms of growth and aging, especially during phases of high oxidative stress that result in molecular damage. Changes affecting lipid class or fatty acid (FA) compositions could affect phospholipid (PL) properties and alter mitochondrial function. In the present study, mitochondrial membrane PL compositions were analysed throughout the life-cycle of Nothobranchius furzeri, a species with explosive growth and one of the shortest-lived vertebrates. Mitochondrial PLs showed several changes with age. Proportions of total PLs and PC were reduced while an increase in PS, CL and PE was observed, mainly between the 2.5 and 5 months of fish age, the time during which animals doubled their weight. FA compositions of individual PLs in mitochondria were also significantly affected with age suggesting the existence of increasing damage to mitochondrial lipids during the life-cycle of N. furzeri that could be one of the main contributors to degraded mitochondrial function associated with aging. The peroxidation index values from N. furzeri mitochondrial PLs were significantly lower than those reported in N. rachovii, a species with a twofold longer life span than N. furzeri, which seems to contradict the membrane pacemaker theory of animal metabolism.

Effects of global change factors on fatty acids and mycosporine-like amino acid production in Chroothece richteriana (Rhodophyta).

Under natural conditions, Chroothece richteriana synthesizes a fairly high proportion of fatty acids. However, nothing is known about how environmental changes affect their production, or about the production of protective compounds, when colonies develop under full sunshine with high levels of UV radiation. In this study, wild colonies of C. richteriana were subjected to increasing temperature, conductivity, ammonium concentrations and photosynthetically active radiation (PAR), and UV radiations to assess the potential changes in lipid composition and mycosporine-like amino acids (MAAs) concentration. The PERMANOVA analysis detected no differences for the whole fatty acid profile among treatments, but the percentages of α-linolenic acid and total polyunsaturated fatty acids increased at the lowest assayed temperature. The percentages of linoleic and α-linolenic acids increased with lowering temperature. γ-linolenic and arachidonic acids decreased with increasing conductivity, and a high arachidonic acid concentration was related with increased conductivity. The samples exposed to UVB radiation showed higher percentages of eicosapentaenoic acid and total monounsaturated fatty acids, at the expense of saturated fatty acids. MAAs accumulation increased but not significantly at the lowest conductivity, and also with the highest PAR and UVR exposure, while ammonium and temperature had no effect. The observed changes are probably related with adaptations of both membrane fluidity to low temperature, and metabolism to protect cells against UV radiation damage. The results suggest the potential to change lipid composition and MAAs concentration in response to environmental stressful conditions due to climate change, and highlight the interest of the species in future research about the biotechnological production of both compound types.

EFFECT OF LONG TERM INTAKE OF WHITE TEA ON ACUTE OXIDATIVE STRESS IN RATS.

INTRODUCTION: the well known antioxidant properties of white tea include the prevention of cancer, neurodegenerative diseases and oxidative stress. Adriamycin can generate an amount of oxidative stress in vivo. OBJECTIVE: evaluate long term intake of white tea on plasma antioxidant capacity and on the fatty acid profile of liver and heart microsomes in animals subjected to acute oxidative stress. METHODS: rats were given distilled water (controls), 15 mg/d (dose 1) or 45 mg/d (dose 2) of solid white tea extract/per kilogram of body weight for 12 months. After this time, all the animals received an injection of adriamycin (ADR) (10 mg/kg body weight), except half of the control group, which were given an injection of saline solution. Samples of plasma and liver and heart were taken. The antioxidant activity, the carbonyl groups and hydroperoxide concentration were analyzed in plasma, and the fatty acid profiles of liver and heart microsomes were obtained. RESULTS & DISCUSSION: only the hydroperoxides showed significant changes, while slight tendencies were observed in antioxidant activity and the carbonyl groups. Although the long term intake of white tea and the administration of adriamycin did not change the fatty acid profile, slight tendencies existed for the SFAs, MUFAs and PUFAs.

Introducción: las propiedades antioxidantes del té son ampliamente conocidas, entre las que se incluyen la prevención del cáncer, diversas enfermedades neurodegenerativas y otras patologías relacionadas con el estrés oxidativo. Por otro lado, la adriamicina es un agente antitumoral que tiene la capacidad de generar estrés oxidativo in vivo. Objetivo: valorar el efecto de la ingesta de té blanco a largo plazo sobre la capacidad antioxidante plasmática y el perfil de ácidos grasos de microsomas de hígado y corazón en animales sometidos a estrés oxidativo agudo. Métodos: se dispuso de ratas a las que se les administró diferentes dosis de té blanco: 0,15 y 45 mg de extracto sólido de té/kg de peso corporal durante 12 meses. Tras este periodo de tratamiento con té blanco, todos los animales recibieron una inyección intraperitoneal de adriamicina (ADR), 10 mg/kg de peso corporal, excepto la mitad del grupo control, que recibieron una inyección de solución salina. Fueron obtenidas muestras de sangre, corazón e hígado. Se analizó la capacidad antioxidante total y se realizaron análisis de oxidación proteica y lipídica en plasma. Además, se obtuvo la fase microsomal de hígado y corazón. Resultado y discusión: se observó una fuerte oxidación lipídica en plasma y una recuperación en los animales tratados con las diferentes dosis de té. La actividad antioxidante y la oxidación proteica, aunque relevantes, solo muestran una ligera tendencia a recuperarse con el tratamiento con té. En cuanto al perfil de ácidos grasos, solamente se observan ligeras tendencias en el porcentaje de ácidos grasos saturados, monoinsaturados y poliinsaturados.

The freshwater alga Chroothece richteriana (Rhodophyta) as a potential source of lipids.

During an ecological study of Chroothece (Rhodophyta) in a small river in a semi-arid region of south-east Spain it became clear that most of these cells had a high lipid content. This suggested potential uses in biotechnology, which has been investigated further. The colonies, which occur in full sunlight, are typically orange-brown. Most, perhaps all, the yellow-orange colour is associated with their high carotenoid content, with the carotenoid to chlorophyll ratio up to 2.7. The polyunsaturated fatty acyl composition of the glycerides was 35.3% of the dry weight. This consisted mainly of omega-3 (5.9%) and omega-6 (29.4%) fats. The relatively high proportion of docosahexaenoyl (1.78%), eicosapentaenoyl (14.15%), arachidonoyl (0.92%) and γ-linolenoyl (0.78%) suggests use for medical and dietary purposes. All cells have a high phycocyanin content whilst phycoerythrin is absent. The alga has a wide distribution globally and hence provides scope for selecting strains with optimum properties.

[Characterization and nutritional value of a food artisan: the meat pie of Murcia]. / Caracterización y valor nutritivo de un alimento artesanal; el pastel de carne de Murcia.

OBJECTIVES: The main aims of this study are to describe the characteristics of the meat pie, a typical product of the regional gastronomy of Murcia and to determine its nutritional and energy values, fatty acid profile and fat quality. METHODS: There were studied 24 samples of Murcia's meat pie from the six best-selling retail establishments in this Region (four units per establishment).The moisture, protein, fat, carbohydrates, fibre and minerals, contents and the energy value, fatty acid profile and fat quality were analyzed using the Official Analysis Methods of Foods. All analyses were performed by triplicate. RESULTS AND DISCUSSION: The average weight of this product was 192.3 ± 11.8 g, with three differentiated parts (base, filling and pastry lid). All ingredients were natural raw materials: wheat flour, lard, ground beef, sliced boiled egg and chorizo, water and spices (salt, pepper, garlic, paprika and nutmeg). Most of its organoleptic attributes are due to the type and amount of fat or lard. The combination of the other ingredients and the particular formulation of the spices are also responsible of other attributes. Due to its protein content (11.0%), this meat pie can replace other meat dishes, and be incorporated into a balanced diet. However, it is necessary to take into account its fat and energy contents (17.3 g and 317 kcal/100 g, respectively). Unlike many common pastry products, it contains no trans fatty acids. CONCLUSIONS: The results of the research show that the studied product remains, at present, an artisan food, and offer reliable information that it is representative of the energy and nutritional values of the Murcia's meat pie, a typical product of the gastronomy of the Region of Murcia.

Objetivos: Describir las características de un producto típico de la gastronomía murciana, determinar su valor nutritivo y energético, analizar su perfil de ácidos grasos y la calidad de su grasa. Material y métodos: Se estudiaron 24 muestras de Pastel de carne de Murcia, procedentes de los seis establecimientos de mayor venta en la Región. (Cuatro unidades por establecimiento). Mediante los métodos oficiales de análisis de alimentos se determinaron los contenidos de humedad, proteína, grasa, carbohidratos, fibra y minerales, el valor energético, el perfil de ácidos grasos (Método de Folch et al. 1957) y la calidad de la grasa. Todos los análisis se realizaron por triplicado. Resultados y discusión: El peso medio fue de 192,3 ± 11,8 g, y en él se diferenciaron tres partes (base, relleno y tapa de hojaldre). Todos sus ingredientes fueron materias primas naturales: harina de trigo, manteca de cerdo, carne picada de ternera, rodajas de huevo cocido y de chorizo, agua y especias (sal, pimienta, ajo, pimentón y nuez moscada). La mayoría de sus atributos organolépticos son debidos al tipo y cantidad de la grasa, manteca de cerdo. El resto de atributos los aporta la combinación de los otros ingredientes y particularmente la formulación de las especias. Por su contenido en proteínas (11,0%), este producto puede sustituir a otros platos a base de carne, e incorporarlo a una dieta equilibrada, pero teniendo en cuenta su contenido en grasa (17,3%) y su aporte energético (317 kcal/100 g). A diferencia de otros muchos productos actuales de pastelería, éste no contiene ácidos grasos trans. Conclusiones: El estudio revela que el producto analizado sigue siendo, en el momento actual, un alimento artesanal, y ofrece información fiable y representativa del valor nutritivo y energético del pastel de carne de Murcia, un producto típico de la gastronomía de la Región de Murcia.

Effect of the consumption of a fruit and vegetable soup with high in vitro carotenoid bioaccessibility on serum carotenoid concentrations and markers of oxidative stress in young men.

AIM: To evaluate the effect of the daily intake of a fruit & vegetable soup with high in vitro bioaccessibility of carotenoids on ß-carotene and lycopene serum concentrations. METHODS: Fourteen healthy young men (24 ± 1 years) received 300 mL/day of a carrot, tomato, and broccoli soup, containing 3.9 mg ß-carotene and 4 mg lycopene, for 4 weeks followed by a 4-week washout period. The serum carotenoid response and oxidative markers were analyzed after 3 and 4 weeks of soup consumption and after a 4-week washout. RESULTS: The in vitro bioaccessibility of ß-carotene and lycopene was 55 and 43%, respectively, in the soup. Serum ß-carotene concentrations were significantly higher than baseline (0.33 ± 0.05 µmol/L) after 3 weeks (0.69 ± 0.06 µmol/L) and 4 weeks (0.78 ± 0.10 µmol/L) of soup consumption (P < 0.001). Serum lycopene was also significantly higher compared with baseline levels (0.26 ± 0.08-0.56 ± 0.04 µmol/L and 0.60 ± 0.04 µmol/L, after 3 and 4 weeks, respectively) (P < 0.001). Although the highest concentration of both carotenoids was found after 4 weeks, the levels were not statistically different from the levels at 3 weeks. A 4-week washout significantly decreased serum carotenoid concentrations, although only ß-carotene returned to baseline. Glutathione peroxidase (GPx) increased significantly after soup supplementation compared with baseline, while superoxide dismutase was significantly lower only after 3 weeks. Glutathione reductase, lipid, protein, and DNA oxidative markers remained unchanged. CONCLUSIONS: The soup contributed to increasing the concentration of each carotenoid by more than 100% after 3 and 4 weeks of consumption, the maximum increase being observed after 4 weeks. Oxidative markers did not show any variation except for GPx. Serum lycopene half-life was longer than that of ß-carotene, which may be important for studies evaluating both carotenoids.

Cell-based assay to quantify the antioxidant effect of food-derived carotenoids enriched in postprandial human chylomicrons.

We developed a new method to evaluate the antioxidant effect of food products in a biological system. The antioxidant status of HepG2 cells was quantified after incubation with postprandial human chylomicrons after the intake of vegetable products. Three subjects consumed in a meal a vegetable soup containing 8.4 mg of ß-carotene and 9 mg of lycopene. After 5 h, the subjects consumed a second meal without carotenoids. Blood samples were collected at basal time and every hour for 9 h. Chylomicrons were isolated from serum samples and used for both carotenoid quantification and HepG2 stimulation. Carotenoid in chylomicrons followed an inter-individual and bimodal carotenoid response. We demonstrated the antioxidant effect of postprandial chylomicrons in HepG2 at the time of maximum carotenoid concentration of chylomicrons with respect to basal time. This cell-based assay seems to be a useful method to evaluate the antioxidant effect of fruit and vegetable products in a biological system.