Teleost fish larvae adapt to dietary arachidonic acid supply through modulation of the expression of lipid metabolism and stress response genes

Dietary fatty acid supply can affect stress response in fish during early development. Although knowledge on the mechanisms involved in fatty acid regulation of stress tolerance is scarce, it has often been hypothesised that eicosanoid profiles can influence cortisol production. Genomic cortisol actions are mediated by cytosolic receptors which may respond to cellular fatty acid signalling. An experiment was designed to test the effects of feeding gilthead sea-bream larvae with four microdiets, containing graded arachidonic acid (ARA) levels (0·4, 0·8, 1·5 and 3·0 %), on the expression of genes involved in stress response (steroidogenic acute regulatory protein, glucocorticoid receptor and phosphoenolpyruvate carboxykinase), lipid and, particularly, eicosanoid metabolism (hormone-sensitive lipase, PPARα, phospholipase A2, cyclo-oxygenase-2 and 5-lipoxygenase), as determined by real-time quantitative PCR. Fish fatty acid phenotypes reflected dietary fatty acid profiles. Growth performance, survival after acute stress and similar whole-body basal cortisol levels suggested that sea-bream larvae could tolerate a wide range of dietary ARA levels. Transcription of all genes analysed was significantly reduced at dietary ARA levels above 0·4 %. Nonetheless, despite practical suppression of phospholipase A2 transcription, higher leukotriene B4 levels were detected in larvae fed 3·0 % ARA, whereas a similar trend was observed regarding PGE2 production. The present study demonstrates that adaptation to a wide range of dietary ARA levels in gilthead sea-bream larvae involves the modulation of the expression of genes related to eicosanoid synthesis, lipid metabolism and stress response. The roles of ARA, other polyunsaturates and eicosanoids as signals in this process are discussed.

Effect of Temperature on the Development and Survival of the Argentine Ant, Linepithema humile

The influence of temperature on the developmental times and survival of insects can largely determine their distribution. For invasive species, like the Argentine ant, Linepithema humile Mayr (Hymenoptera: Formicidae), these data are essential for predicting their potential range based on mechanistic models. In the case of this species, such data are too scarce and incomplete to make accurate predictions based on its physiological needs. This research provides comprehensive new data about brood survival and developmental times at a wide range of temperatures under laboratory conditions. Temperature affected both the complete brood development from egg to adult worker and each of the immature stages separately. The higher the temperature, the shorter the development times. Brood survival from egg to adult was low, with the maximum survival rate being only 16% at 26º C. Temperature also affected survival of each of the immature stages differently: eggs were negatively affected by high temperatures, while larvae were negatively affected by low temperatures, and the survival of pupae was apparentlyindependent of environmental temperature. At 32º C no eggs survived, while at 18º C less than 2% of the eggs hatched into larva. The data from the present study are essential for developing prediction models about the distribution range of this tramp species based on its physiological needs in relation to temperature

Effect of Temperature on the Development and Survival of the Argentine Ant, Linepithema humile

The influence of temperature on the developmental times and survival of insects can largely determine their distribution. For invasive species, like the Argentine ant, Linepithema humile Mayr (Hymenoptera: Formicidae), these data are essential for predicting their potential range based on mechanistic models. In the case of this species, such data are too scarce and incomplete to make accurate predictions based on its physiological needs. This research provides comprehensive new data about brood survival and developmental times at a wide range of temperatures under laboratory conditions. Temperature affected both the complete brood development from egg to adult worker and each of the immature stages separately. The higher the temperature, the shorter the development times. Brood survival from egg to adult was low, with the maximum survival rate being only 16% at 26° C. Temperature also affected survival of each of the immature stages differently: eggs were negatively affected by high temperatures, while larvae were negatively affected by low temperatures, and the survival of pupae was apparently independent of environmental temperature. At 32° C no eggs survived, while at 18° C less than 2% of the eggs hatched into larva. The data from the present study are essential for developing prediction models about the distribution range of this tramp species based on its physiological needs in relation to temperature

The implementation of liquid chromatography tandem mass spectrometry for the official control of lipophilic toxins in seafood: Single-laboratory validation under four chromatographic conditions

We performed a comprehensive study to assess the fit for purpose of four chromatographic conditions for the determination of six groups of marine lipophilic toxins (okadaic acid and dinophysistoxins, pectenotoxins, azaspiracids, yessotoxins, gymnodimine and spirolides) by LC-MS/MS to select the most suitable conditions as stated by the European Union Reference Laboratory for Marine Biotoxins (EURLMB). For every case, the elution gradient has been optimized to achieve a total run-time cycle of 12 min. We performed a single-laboratory validation for the analysis of three relevant matrices for the seafood aquaculture industry (mussels, pacific oysters and clams), and for sea urchins for which no data about lipophilic toxins have been reported before. Moreover, we have compared the method performance under alkaline conditions using two quantification strategies: the external standard calibration (EXS) and the matrix-matched standard calibration (MMS). Alkaline conditions were the only scenario that allowed detection windows with polarity switching in a 3200 QTrap mass spectrometer, thus the analysis of all toxins can be accomplished in a single run, increasing sample throughput. The limits of quantification under alkaline conditions met the validation requirements established by the EURLMB for all toxins and matrices, while the remaining conditions failed in some cases. The accuracy of the method and the matrix effects where generally dependent on the mobile phases and the seafood species. The MMS had a moderate positive impact on method accuracy for crude extracts, but it showed poor trueness for seafood species other than mussels when analyzing hydrolyzed extracts. Alkaline conditions with EXS and recovery correction for OA were selected as the most proper conditions in the context of our laboratory. This comparative study can help other laboratories to choose the best conditions for the implementation of LC-MS/MS according to their own necessities.

Some like it hot: temperature and acidification modulate larval development and settlement of the sea urchin Arbacia lixula

We studied the effects of temperature and pH on larval development, settlement and juvenile survival of a Mediterranean population of the sea urchin Arbacia lixula. Three temperatures (16, 17.5 and 19 °C) were tested at present pH conditions (pHT 8.1). At 19 °C, two pH levels were compared to reflect present average (pHT 8.1) and near-future average conditions (pHT 7.7, expected by 2100). Larvae were reared for 52-days to achieve the full larval development and complete the metamorphosis to the settler stage. We analyzed larval survival, growth, morphology and settlement success. We also tested the carry-over effect of acidification on juvenile survival after 3 days. Our results showed that larval survival and size significantly increased with temperature. Acidification resulted in higher survival rates and developmental delay. Larval morphology was significantly altered by low temperatures, which led to narrower larvae with relatively shorter skeletal rods, but larval morphology was only marginally affected by acidification. No carry-over effects between larvae and juveniles were detected in early settler survival, though settlers from larvae reared at pH 7.7 were significantly smaller than their counterparts developed at pH 8.1. These results suggest an overall positive effect of environmental parameters related to global change on the reproduction of A. lixula, and reinforce the concerns about the increasing negative impact on shallow Mediterranean ecosystems of this post-glacial colonizer.

Trichoptera (INSECTA) collected in Mediterranean river basins in the Iberian Peninsula: taxonomic remarks and notes on ecology

As a result of the GUADALMED project, undertaken in Iberian Mediterranean basins, to which were added samples taken by the junior author in the area, we collected numerous caddisfly larvae, pupae and adults. Some larvae were also reared in the lab to obtain adults and allow proper identification. A total of 90 species were identified, which accounts for more than a fourth of the species known in the Iberian Peninsula and Balearic Islands. Here we confirm the presence of doubtful species in the Iberian Peninsula (Glyphotaelius pellucidus) and we expand the distribution range of others (Lype reducta, Micrasema minimum, Limnephilus guadarramicus, Sericostoma pyrenaicum). Moreover, because of the unconformity of morphological larval characteristics with present taxonomical keys (Mesophylax aspersus) or lack of larvae descriptions (Allogamus mortoni, Stenophylax espanioli), here we include some relevant taxonomical aspects that are useful to identify larvae. A brief description of the larva of a possible new species of Hydropsyche (from now on H. gr. instabilis) is also given. Los tricópteros (Insecta) recolectados en las cuencas mediterráneas de la Península Ibérica: notas taxonómicas y requerimientos ecológicos Como resultado de los estudios realizados en el proyecto GUADALMED en las cuencas de los ríos mediterráneos peninsulares y otros muestreos realizados por la primera autora del trabajo se han recolectado numerosas larvas, pupas y adultos de tricópteros. Algunas larvas fueron criadas en el laboratorio para la obtención de adultos y con ello asegurar su identificación. Se han identificado un total de 90 especies que suponen más de una cuarta parte de las especies actualmente conocidas en la Península Ibérica y Baleares. En este trabajo se confirma la presencia, hasta ahora dudosa en la Península Ibérica, de algunas especies (Glyphotaelius pellucidus) y se amplia el área de distribución de otras (Lype reducta, Micrasema minimum, Limnephilus guadarramicus, Sericostoma pyrenaicum). Además, se incluyen algunos aspectos taxonómicos relevantes para la identificación de algunas larvas en futuros estudios, debido a que las claves ya existentes no describen correctamente la larva (Mesophylax aspersus) o porque se trata de larvas sin describir (Allogamus mortoni, Stenophylax espanioli). Se añade también una somera descripción de la larva de una posible nueva especie de Hydropsyche (citada aquí como H. gr. instabilis) y que requiere un estudio más detallado.

Mitochondrial control region and microsatellite data show low genetic variability in reared senegalese sole, Solea senegalensis. Preliminary data

Peer reviewed

LRCW.NET: A web site with a virtual laboratory for the study of coarse and cooking wares in the Late Antique Mediterranean

This contribution presents the LRCW.net a website with a virtual laboratory intranet devoted to the study of coarse and cooking wares in the late Antique Mediterranean. It is designed as a public website with a virtual laboratory intranet. There, all institutions and researchers interested in the subject can work together towards a specific purpose such as the creation of an on-line ‘encyclopedia’ for these categories of ceramics. The LRCW.net website and the associated virtual laboratory are just a small part of a wider initiative that aims to create an on-line Encyclopedia for Ancient Ceramics in the Mediterranean.

Laboratory diagnosis of brucellosis in a rural endemic area in northeastern Spain

Sera obtained from 62 patients from four mountain counties in Catalonia (Northeastern Spain), in whom brucellosis had been diagnosed on the basis of clinical evidence and/or personal history, were analyzed using the rose Bengal test, standard serum agglutination test (SAT), Coombs" test, ELISA, and complement fixation. The diagnosis was further confirmed through blood cultures. Clinical evidence, epidemiology, and the results from serologic tests were used to assign patients to one of two groups: group 1 (n = 38) patients had primary infections, whereas group 2 (n = 24) patients had been previously exposed to the microorganism, i.e. re-infection of group 2 individuals occurred after long periods of time during which no active infection by Brucella had been detected. Receivingoperating charts (ROC) were used to determine the diagnostic value of the different tests and to establish discriminant values. Blood culture was a valuable diagnostic tool in group 1 (0.92 sensitivity) but was inappropriate in group 2 (0.08). The combination of positive rose Bengal test and agglutination ≥1/160 was valid for diagnosis in group 1. In group 2, agglutination < 1/160 (including negative agglutination) did not rule out brucellosis. The combination of positive rose Bengal test and Coombs" test ≥1/320 was the best diagnostic criterion (0.8 specificity; 1 sensitivity). ELISA (for IgG, IgM, or both) did not improve diagnostic accuracy

Adult sampling and larva rearing of the cork oak pest Coraebus undatus (Coleoptera: Buprestidae)

Coraebus undatus is the main insect pest of cork oak worldwide. The larvae tunnel in the cortical cambium filling the bark with galleries and causing the cork to break at harvest. The first objective of this study was to test the effect of purple traps in the attraction of C. undatus because this colour is attractive to other buprestid beetles. The second objective was to develop a diet in which field-collected larvae could be reared to adulthood. Pairs of purple and clear (control) sticky traps were placed in a cork oak forest in Girona, Spain in the summer of 2008

Desarrollo larvario y número de estadios larvarios de "Pseudaletia unipuncta" alimentada con dos variedades de maíz y dos dietas semisintéticas

Pseudaletia ( Mythimna) unipuncta (Haworth) es una plaga defoliadora del maíz presente año tras año en la Cuenca del Ebro, que realiza en el maíz daños esporádicos pero a veces devastadores. La implantación creciente de maíz resistente a taladros ha producido, en la zona del presente estudio, la disminución casi total de los daños debidos a los taladros Sesamia nonagrioides (Lefebvre) y Ostrinia nubilalis (Hübner). Sin embargo, no se sabe aún cómo el incremento de maíz transgénico puede afectar a las poblaciones de P. unipuncta. Actualmente no hay datos sobre biología o parámetros del desarrollo de la especie en la Península Ibérica necesarios para evaluar los posibles efectos de las nuevas variedades utilizadas. Para ello es necesario primero establecer un método de cría adecuado y posteriormente iniciar los estudios sobre la biología de la misma. En el presente trabajo se planteó conocer diferentes parámetros del desarrollo larvario de poblaciones locales de P. unipuncta alimentada sobre hoja de maíz (variedades Tietar y PR33P66) y sobre dos dietas semisintéticas, una a base de alubias. Phaseolus vulgaris, y otra a base de maíz, Zea maya. Los resultados mostraron que las larvas desarrolladas en dieta a base de alubias sufrieron menor mortalidad, que las desarrolladas con planta o sobre la otra dieta, además el peso de las pupas resultantes fue mayor y presentaron menos malformaciones. En cuanto al numero de estadios larvarios, la mayoría pupó tras seis mudas larvarias, necesitando, las de dieta de maíz, estadios larvarios adicionales. Estos resultados indican que la dieta a base de alubias es la más adecuada para posteriores trabajos con esta especie.

Assessment of insecticide resistance in eggs and neonate larvae of Cydia pomonella (Lepidoptera: Tortricidae)

Spanish Cydia pomonella (L.) field populations have developed resistance to several insecticide groups. Diagnostic concentrations were established as the LC90 calculated on a susceptible strain (S_Spain) for five and seven insecticides and tested on eggs and neonate larvae field populations, respectively. The three most relevant enzymatic detoxification systems (mixed-function oxidases (MFO), glutathione S-tranferases (GST) and esterases (EST)) were studied for neonate larvae. In eggs, 96% of the field populations showed a significantly lower efficacy when compared with the susceptible strain (S_Spain) and the most effective insecticides were fenoxycarb and thiacloprid. In neonate larvae, a significant loss of susceptibility to the insecticides was detected. Flufenoxuron, azinphos-methyl and phosmet showed the lowest efficacy, while lambda-cyhalothrin, alpha-cypermethrin and chlorpyrifos-ethyl showed the highest. Biochemical assays showed that the most important enzymatic system involved in insecticide detoxification was MFO, with highest enzymatic activity ratios (5.1–16.6 for neonates from nine field populations). An enhanced GST and EST activities was detected in one field population, with enzymatic activity ratios of threefold and fivefold for GST and EST, respectively, when compared with the susceptible strain. The insecticide bioassays showed that the LC90 used were effective as diagnostic concentrations. Measures of MFO activity alongside bioassays with insecticide diagnostic concentrations could be used as tools for monitoring insecticide resistance in neonate larvae of C. pomonella.

Toxicity of allyl esters in insect cell lines and in Spodoptera littoralis larvae

We investigated the effects of five allyl esters, two aromatic (allyl cinnamate and allyl 2-furoate) and three aliphatic (allyl hexanoate, allyl heptanoate, and allyl octanoate) in established insect cell lines derived from different species and tissues. We studied embryonic cells of the fruit fly Drosophila melanogaster (S2) (Diptera) and the beet armyworm Spodoptera exigua (Se4) (Lepidoptera), fat body cells of the Colorado potato beetle Leptinotarsa decemlineata (CPB) (Coleoptera), ovarian cells of the silkmoth Bombyx mori (Bm5), and midgut cells of the spruce budworm Choristoneura fumiferana (CF203) (Lepidoptera). Cytotoxicity was determined with use of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and trypan blue. In addition, we tested the entomotoxic action of allyl cinnamate against the cotton leafworm Spodoptera littoralis .The median (50%) cytotoxic concentrations (EC50s) of the five allyl esters in the MTT bioassays ranged between 0.25 and 27 mM with significant differences among allyl esters (P = 0.0012), cell lines (P < 0.0001), and the allyl estercell line interaction (P < 0.0001). Allyl cinnamate was the most active product, and CF203 the most sensitive cell line. In the trypan blue bioassays, cytotoxicity was produced rapidly and followed the same trend observed in the MTT bioassay. In first instars of S. littoralis, allyl cinnamate killed all larvae at 0.25% in the diet after 1 day, while this happened in third instars after 5 days. The LC50 in first instars was 0.08%. In addition, larval weight gain was reduced (P < 0.05) after 1 day of feeding on diet with 0.05%. In conclusion, the data provide evidence of the significant but differential cytotoxicity among allyl esters in insect cells of different species and tissues. Midgut cells show high sensitivity, indicating the insect midgut as a primary target tissue. Allyl cinnamate caused rapid toxic effects in S. littoralis larvae at low concentrations, suggesting further potential for use in pest control.

Standardisation of elemental analytical techniques applied to provenance studies of archaeological ceramics: an inter laboratory calibration study

Chemical analysis is a well-established procedure for the provenancing of archaeological ceramics. Various analytical techniques are routinely used and large amounts of data have been accumulated so far in data banks. However, in order to exchange results obtained by different laboratories, the respective analytical procedures need to be tested in terms of their inter-comparability. In this study, the schemes of analysis used in four laboratories that are involved in archaeological pottery studies on a routine basis were compared. The techniques investigated were neutron activation analysis (NAA), X-ray fluorescence analysis (XRF), inductively coupled plasma optical emission spectrometry (ICP-OES) and inductively coupled plasma mass spectrometry (ICP-MS). For this comparison series of measurements on different geological standard reference materials (SRM) were carried out and the results were statistically evaluated. An attempt was also made towards the establishment of calibration factors between pairs of analytical setups in order to smooth the systematic differences among the results.