A rapid latex agglutination test for gender identification in the Atlantic bluefin tuna,Thunnus thynnus (Linnaeus)

A rapid, one‐step agglutination assay has been developed, based on latex particles sensitized with antibodies against vitellogenin (Vtg), aimed at Atlantic bluefin tuna, Thunnus thynnus (Linnaeus) (ABFT), gender identification. The egg‐yolk precursor protein Vtg was used as a gender marker for the assay as it is a female‐specific protein synthesized during reproductive maturation. The presence of Vtg in the plasma was revealed in 60–120 s through an agglutination reaction by mixing small volumes of ABFT plasma and an anti‐Vtg antibody‐latex suspension on a microscope slide. The effectiveness of the present test was restricted to the months of May and June, concomitant with high circulating Vtg levels. Because of its rapidity and ease of performance in the field, the present gender identification assay could be useful for broodstock management in the aquaculture industry as well as in tagging studies on wild populations.     

The effects of contrasted deficit irrigation strategies on the fruit growth and kernel quality of mature almond trees

The aim of this study was to quantify and compare the effects of two different deficit irrigation (DI) strategies (regulated deficit irrigation, or RDI, and partial rootzone drying, PRD) on almond (Prunus dulcis (Mill.) D.A. Webb) fruit growth and quality. Five irrigation treatments, ranging from moderate to severe water restriction, were applied: (i) full irrigation (FI), irrigated to satisfy the maximum crop water requirements (ET_c); (ii) regulated deficit irrigation (RDI), receiving 50% of ET_c during the kernel-filling stage and at 100% ET_c throughout the remaining periods; and three PRD treatments – PRD₇₀, PRD₅₀ and PRD₃₀ – irrigated at 70%, 50% and 30% ET_c, respectively, during the whole growth season. The DI treatments did not affect the overall fruit growth pattern compared to the FI treatment, but they had a negative impact on the final kernel dry weight for the most stressed treatments. The allocation of water to the different components of the fruit, characterized by the fresh weight ratio of kernel to fruit, appeared to be the process most clearly affected by DI. Attributes of the kernel chemical composition (lipid, protein, sugar and organic acid contents) were not negatively affected by the intensity of water deprivation. Overall, our results indicated that PRD did not present a clear advantage (or disadvantage) over RDI with regard to almond fruit growth and quality.     

Dietary lipid sources for seabream and seabass: growth performance, tissue composition and flesh quality

Due to its traditionally good availability, digestibility and high content of n ? 3 HUFA, fish oil is the main lipid source in fish feeds. However, world demand for this product has grown significantly in recent years, whereas its production, based on fisheries landings, is static. The purpose of the present study was to assess the effect of partial replacement of fish oil in compound diets for gilthead seabream and seabass, by several vegetable oil sources, on growth, dietary fatty acid utilization and flesh quality. Five iso‐energetic and isoproteic experimental diets were formulated (25% lipid content). Fish oil was the only added lipid source in the control (FO) diet, and it was included in the other experimental diets at a level high enough (40% of FO diet) to keep the n ? 3 HUFA levels well over 3% in order to cover the essential fatty acid requirements of these species. Fish oil was replaced by soyabean oil (SO), rapeseed oil (RO) and linseed oil (LO) or a mixture (Mix) of them. Feed intake in all dietary groups was in the range of results obtained for commercial diets in both species, and growth and feed utilization were very good. The results show that, providing a minimum content of essential fatty acids in the diet, it is possible to replace up to 60% of the fish oil by SO, LO and RO or a mixture of them in diets for seabream and seabass, without compromising fish growth. Fatty acid composition of liver and muscle reflected that of the diet, but utilization of dietary lipids differed between these two tissues and was also different for the different fatty acids. Despite reduction in dietary saturated fatty acids by the inclusion of vegetable oils, their levels in fish liver were as high as in fish fed the fish oil diet, whereas, in muscle, levels were reduced according to that in the diet. Linoleic and linolenic acids were accumulated in the liver proportionally to their levels in the diet, suggesting a lower oxidation of these fatty acids in comparison to other 18C fatty acids. Regarding eicosapentaenoic acid (20 : 5n ? 3; EPA), docosahexaenoic acid (22 : 6n ? 3; DHA) and arachidonic acid (20 : 4n ? 6; ARA), these essential fatty acids were reduced in the liver at a similar rate, whereas DHA was preferentially retained in the muscle in comparison with the other fatty acids, denoting a higher oxidation particularly of EPA, in the muscle. Some other PUFA increased despite their low dietary levels in seabream fed LO diets and in seabass fed SO diet, suggesting the stimulation of delta‐6 and delta‐5 desaturase activity in marine fish. Despite differences in fatty acid composition, fillet of fish fed vegetable oils was very well accepted by trained judges when assessed cooked.     

High stocking density produces crowding stress altering some physiological and biochemical parameters in gilthead seabream, Sparus aurata, juveniles

High stocking density has been shown to produce a wide variety of effects on cultured fish populations, such as alterations in behavior and poor feed utilization, resulting in mortality and poor growth. High stocking density has also been reported to produce chronic stress and mobilization of energy sources in fish. There are few studies focusing on the effect of high stocking density on one of the most important marine fish species for Mediterranean aquaculture, gilthead seabream (Sparus aurata). Consequently, this study investigates the effect of high stocking density on juveniles of this species, focusing on effects of stocking density on growth, biochemical composition, immune status and hematology.The results of this study showed that high stocking density produced a chronic stress situation. Plasma cortisol levels increased up to 16.25 ng ml–1 in fish held at high stocking; this value was 4 fold that of fish held at low stocking; density (3.91 ng ml–1 ). As a consequenceof this stressful situation, there was both haemoconcentration and a decrease in alternative complement pathway (ACP), an important component of the immune system of fish (from 167.23 U ml–1 down to 146.37 U ml–1). Haematocrit, haemoglobin concentration and red blood cell count were significantly higher in fish held at high stocking density (43.87%, 10.76 9dl–1 and 3.36 ×106 mm–3, respectively) compared with those fish held at low stocking density (37.21%, 9.32 g/dlg–1 and 2.82 ×106 mm–3, respectively).In addition, high stocking density produced a decrease in hepatosomatic index (from 2.26 down to 2.04) and altered liver fatty acid composition. Oleic acid (18: 1n-9) decreased in liver total lipids of fish held at high stocking density and arachidonic acid (20: 4-n6) and n-3 high unsaturated fatty acids (n-3 HUFA) were reduced in liver polar lipids of those fish. These alterations reflect the effect of stocking density on lipid metabolism to help meet the increased energy demand.     

Computed tomographic‐lymphography as a complementary technique for lymph node staging in dogs with malignant tumors of various sites

Locoregional lymph nodes are routinely examined in order to define the spatial extent of neoplastic disease. As draining patterns of certain tumor types can be divergent from expected anatomical distribution, it is critical to sample the lymph nodes truly representing the draining area. The aim of this bicenter prospective pilot study was to describe the technique of computed tomographic (CT)‐lymphography for primary draining lymph node mapping in tumor staging in dogs. Forty‐five dogs with macro‐ or microscopic tumors in specified localizations were evaluated. Depending on body weight, 0.8–2 ml contrast agent (iohexol) was injected into four quadrants around the tumor, and CT‐images were obtained at 1, 3, 6, 9, and 12 minutes post‐injection. Attenuation of chosen regions of interest (Hounsfield units (HU)) and patterns of enhancement were assessed for 284 lymph nodes in the precontrast study with median HUs of 31.1 (Interquartile range (IQR) = 18.4) and for 275 in the intravenous postcontrast study with 104.3 HU (IQR = 31.2) (paired Wilcoxon test, P < 0.001). In the CT‐lymphography study, 45 primary draining lymph nodes with a significantly higher median HU value of 348.5 (IQR = 591.4) (one‐sample t‐test, P < 0.001) were identified. Primary draining lymph nodes were found to be clearly visible after 1–3 minutes after local injection, often concurrent with a good visibility of the lymphatic vessel system. The herein described technique of peritumorally injected CT‐contrast agent followed by subsequent CT‐lymphography for primary draining lymph node mapping works well in a majority of cases in all investigated sites and warrants further validation for different tumor entities.     

The importance of rice fields for glossy ibis (Plegadis falcinellus): Management recommendations derived from an individual-based model

Artificial wetlands provide alternative habitats for waterbirds. The Doñana rice fields (SW Spain) are extensively used as a foraging site by the glossy ibis (Plegadis falcinellus). The aim of this study was to develop an individual-based model to predict the possible effects of glossy ibis’ population growth, reductions in the rice cultivated area, and changes on the phenology of the management processes of the paddies on the mortality rate of the glossy ibis population. We test the hypothesis that the glossy ibis breeding population of Doñana can obtain its energy requirements during the non-breeding season by feeding on rice fields alone. Our results show that the glossy ibis population growth is not currently limited by rice field availability. However, a reduction of 80% would cause mortality rate increases above the observed mortality (5.9% per year), with values around 10% per year for populations between 50,000 and 100,000 birds. A reduction of 90% of the rice field area would cause mortality rate increase above the observed value for populations greater than 20,000 birds, reaching 60% per year with a population of 100,000 birds. Cultivated area in Doñana suffers temporary reduction on its area during drought periods. Taking into account the fact that the glossy ibis population for 2011 may exceed 22,900 birds, large scale changes in the area of rice fields due to habitat transformations and/or drought periods may have important effects on the viability of the glossy ibis population in Doñana.     

Characterization of carrot root oil arising from supercritical fluid carbon dioxide extraction

Carrot root oil (SCO), obtained by supercritical fluid carbon dioxide (SC-CO2) extraction, was characterized and compared to a commercial carrot oil (MCO) and a virgin olive oil (VOO) (cv. Coratina). SCO showed much higher contents of carotenes, phenolics, waxes, phytosterols, and sesquiterpene and monoterpene volatiles. In SCO, the most prominent components present in the fully investigated analytical fractions (fatty acids, triglycerides, waxes, phytosterols, long-chain aliphatic alcohols, superior triterpene alcohols, and volatiles) were, respectively, linolenic acid, trilinolein, waxes C38, beta-sitosterol, campesterol and stigmasterol, 1-hexacosanol, 24-methylencycloartanol and cycloartenol, beta-caryophyllene, alpha-humulene, alpha-pinene, and sabinene. In VOO, the major constituents of the above analytical classes were, respectively, oleic acid, trilinolein, waxes C36, unsaturated volatile C6 aldehydes (trans-2-hexenal most markedly), and the same prominent sterols and superior alcohols found in SCO. In MCO, which also contained a proportion of unknown plant oil, several components showed magnitudes that were lower compared to SCO but higher with respect to VOO. The last had the aliphatic and triterpene alcohol concentration higher compared to that of both SCO and MCO. Several chemometric methods, applied to different analytical data sets, proved to be effective in grouping the three oil kinds.     

Supercritical CO2 extraction of beta-carotene from a marine strain of the cyanobacterium Synechococcus species

Dynamic extraction of carotenoids from a marine strain of Synechococcus sp. (Cyanophyceae) with supercritical CO2 (SC-CO2) was investigated with regard to operation pressure and temperature effects on extraction efficiency. Extraction yield (milligrams of pigment per gram of dry weight) for SC-CO2) was compared with the extraction yield for dimethylformamide (DMF). Carotenoids extracted with SC-CO2 were beta-carotene (Ct), zeaxanthin (Z), beta-cryptoxanthin (Cr), and equinenone; chlorophyll a was poorly extracted, whereas myxoxanthophyll, another major carotenoid, was not extracted under any experimental condition. The highest relative yield, which is defined here as y(r) = [(mg of pigment(SC-CO2)/mg of pigment(DMF))] x 100, was 76.1 +/- 8.6% for Ct, but it rose to 87.0 +/- 3.4% when 15% ethanol was used as cosolvent. The pressure effect on y(r) was found to be significant (p < 0.05) for both Cr and Z, along with total carotenoids, whereas the effect of square T (TT) was significant for only Ct. From empirical correlations, pairwise pressure (bar) and temperature (degrees C), respectively, for optimal extraction were determined to be (358, 50) for Ct, (454, 59) for Cr, and (500, 60) for Z. Cell disruption by sonication or detergent treatment of the biomass did not improve the extraction efficiency. Matrix structure together with material state could explain the low carotenoid extraction yield obtained with SC-CO2 as compared to DMF in Synechococcus sp. However, the process can be applied to selective extraction of different carotenoids.     

Silver-catalyzed C-C bond formation between methane and ethyl diazoacetate in supercritical CO₂

Even in the context of hydrocarbons' general resistance to selective functionalization, methane's volatility and strong bonds pose a particular challenge. We report here that silver complexes bearing perfluorinated indazolylborate ligands catalyze the reaction of methane (CH(4)) with ethyl diazoacetate (N(2)CHCO(2)Et) to yield ethyl propionate (CH(3)CH(2)CO(2)Et). The use of supercritical carbon dioxide (scCO(2)) as the solvent is key to the reaction's success. Although the catalyst is only sparingly soluble in CH(4)/CO(2) mixtures, optimized conditions presently result in a 19% yield of ethyl propionate (based on starting quantity of the diazoester) at 40°C over 14 hours.     

Distribution and eco-geographic characterization of Carica papaya L. native to Mexico

Genetic Resources and Crop Evolution - Carica papaya L. is an important plant genetic resource that has a low level of genetic erosion. Knowing the distribution and eco-geography of native C....