Centro IFAPA

Cadiz, Spain

Centro IFAPA

Cadiz, Spain
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The effects, in greenhouse conditions, of Purified Urban Wastewater (PW) from Almería (Spain), in the fertigation of pepper (Capsicum annuum L.) on sandy mulch soil, were evaluated. Primary, secondary (active sludges) and tertiary (Chlorination + ozonation) purification treatments were applied to wastewater. Irrigation treatments applied were PW, natural Ground Water (GW), Fertilizer PW (FPW) and Fertilizer GW (FGW). The vegetal biomass, yield and fruit quality were controlled. Heavy metals (Cr, Cd, Pb, Ni, Mn, Cu and Zn), arsenic (As) and Polycyclic Aromatic Hydrocarbons (PAH) in water, soil, leaf, and fruit were analysed. The PW presented heavy metal, As and PAH contents acceptable for its use in drip irrigation. In the soil, fertigated with PW, the concentration of heavy metals and As did not increase, whilst the PAH concentration decreased. The PW treatment supplied enough nutrients to obtain yield and fruit quality equal to that of GW with fertilization. A significant saving on N, P and K fertilizers (37%, 66% and 12% respectively) was achieved by using PW. The Cd, Pb and As contents of the fruit did not show risk for human consumption. The total PAH concentrations in the fruit were low, the highest of which was phenathrene, with no carcinogenic signification.

Scapigliati G.,University of Tuscia | Buonocore F.,University of Tuscia | Randelli E.,University of Tuscia | Casani D.,University of Tuscia | And 19 more authors.
Fish and Shellfish Immunology | Year: 2010

Naïve sea bass juveniles (38.4 ± 4.5 g) were intramuscularly infected with a sublethal dose of betanodavirus isolate 378/I03, followed after 43 days by a similar boosting. This infection resulted in an overall mortality of 7.6%. At various intervals, sampling of fish tissues was performed to investigate: i) B and T lymphocyte content in organs and tissues; ii), proliferation of leucocytes re-stimulated in vitro with inactivated virus; iii) presence of serum antibody specific for betanodavirus; iv) expression of genes coding for the following immunoregulatory molecules involved in innate and acquired responses: type I IFN, Mx, IL-1, Cox-2; IL-10, TGF-β, TCRβ, CD4, CD8α, IgM, by using a quantitative PCR array system developed for sea bass. The obtained results showed a detectable increase of T cells and B cells in PBL during betanodavirus infection. Furthermore, leucocytes obtained from blood, head kidney, and gills showed a detectable "in vitro" increase in viability upon addition of inactivated viral particles, as determined by measuring intracellular ATP concentration. ELISA analysis of sera showed that exposure to nodavirus induced a low, but specific antibody titer measured 43 days after infection, despite the presence of measurable levels of natural antibody. Finally, a strong upregulation of genes coding for type I IFN, Mx, and IgM was identified after both infection and boosting. Interestingly, an upregulation of Cox-2 until boosting, and of TGF-β and IL-10 after boosting was also observed, while the other tested genes did not show any significant variations with respect to mock-treated fish. Overall, our work represents a first comprehensive analysis of cellular and molecular immune parameters in a fish species exposed to a pathogenic virus. © 2009 Elsevier Ltd. All rights reserved.

Merlo M.A.,University of Cádiz | Cross I.,University of Cádiz | Manchado M.,Centro IFAPA | Cardenas S.,Centro IFAPA | Rebordinos L.,University of Cádiz
Journal of Molecular Evolution | Year: 2013

There has been considerable discussion in recent years on the evolution of the tandemly repeated multigene families, since some organisms show a concerted model whereas others show a birth-and-death model. This controversial subject extends to several species of fish. In this study, three species of the Sparidae family (Pagrus pagrus, P. auriga and Diplodus sargus) and an interspecific hybrid (P. pagrus (♀) × P. auriga (â™)) have been studied at both molecular and cytogenetic level, taking three different multigene families (5S rDNA, 45S rDNA and U2 snDNA). Results obtained with the 5S rDNA in P. pagrus and P. auriga are characterized by a considerable degree of conservation at the two levels; however, an extraordinary variation was observed in D. sargus at the two levels, which has never been found in other fishes studied to date. As a consequence of this, the evolutionary model of the multigene families is discussed considering the results obtained and others from the bibliography. The result obtained in the hybrid allowed the recombination frequency in each multigene family to be estimated. © 2013 Springer Science+Business Media New York.

Merlo M.A.,University of Cádiz | Cross I.,University of Cádiz | Chairi H.,University of Cádiz | Manchado M.,Centro IFAPA | Rebordinos L.,University of Cádiz
Genes and Genetic Systems | Year: 2010

By analyzing three multigene families, two closely related and commercially important species, Dicentrarchus labrax and Dicentrarchus punctatus, were characterized by cytogenetic and molecular methods. The interspecies hybrid Dicentrarchus labrax (♀) × Dicentrarchus punctatus (♂) was also analyzed. The multigene families studied were the 5S rDNA, 45S rDNA and the U2 snRNA. A microsatellite GTT motif was found within the non transcribed spacers (NTS) of the 5S rDNA from the two species. However, hexanucleotide duplication next to this microsatellite was observed in the D. labrax and hybrid clones, but not in D. punctatus. The U2 snRNA appeared to be linked to the U5 gene and showed two variant sequences, in both D. labrax and D. punctatus. They differed in one insertion/deletion of 7 nucleotides. The first internal transcribed spacer (ITS-1) region showed higher nucleotide variability in D. punctatus than in D. labrax. Nucleotide polymorphism within species and also nucleotide divergence between species were determined in the different gene regions. In a FISH analysis we obtained three chromosomal markers, because the 5S rDNA, 18S rDNA and U2 snRNA probes hybridized each in three different chromosome pairs. Hence none of them was co-localized. The 5S rDNA cluster and U2 snRNA were localized in acrocentric chromosome pairs, while the 18S rRNA gene probe hybridized in a subtelocentric pair. Finally, the usefulness of the results in developing tools for phylogenetic analysis and species identification are discussed in relation to other fish species.

Merlo M.A.,University of Cádiz | Pacchiarini T.,University of Cádiz | Portela-Bens S.,University of Cádiz | Cross I.,University of Cádiz | And 2 more authors.
BMC Genetics | Year: 2012

Background: Molecular and cytogenetic markers are of great use for to fish characterization, identification, phylogenetics and evolution. Multigene families have proven to be good markers for a better understanding of the variability, organization and evolution of fish species. Three different tandemly-repeated gene families (45S rDNA, 5S rDNA and U2 snDNA) have been studied in Plectorhinchus mediterraneus (Teleostei: Haemulidae), at both molecular and cytogenetic level, to elucidate the taxonomy and evolution of these multigene families, as well as for comparative purposes with other species of the family.Results: Four different types of 5S rDNA were obtained; two of them showed a high homology with that of Raja asterias, and the putative implication of a horizontal transfer event and its consequences for the organization and evolution of the 5S rDNA have been discussed. The other two types do not resemble any other species, but in one of them a putative tRNA-derived SINE was observed for the first time, which could have implications in the evolution of the 5S rDNA. The ITS-1 sequence was more related to a species of another different genus than to that of the same genus, therefore a revision of the Hamulidae family systematic has been proposed. In the analysis of the U2 snDNA, we were able to corroborate that U2 snDNA and U5 snDNA were linked in the same tandem array, and this has interest for tracing evolutionary lines. The karyotype of the species was composed of 2n = 48 acrocentric chromosomes, and each of the three multigene families were located in different chromosome pairs, thus providing three different chromosomal markers.Conclusions: Novel data can be extracted from the results: a putative event of horizontal transfer, a possible tRNA-derived SINE linked to one of the four 5S rDNA types characterized, and a linkage between U2 and U5 snDNA. In addition, a revision of the taxonomy of the Haemulidae family has been suggested, and three cytogenetic markers have been obtained. Some of these results have not been described before in any other fish species. New clues about the genome organization and evolution of the multigene families are offered in this study. © 2012 Merlo et al.; licensee BioMed Central Ltd.

Merlo M.A.,University of Cádiz | Cross I.,University of Cádiz | Rodriguez-Rua A.,Centro IFAPA | Manchado M.,Centro IFAPA | Rebordinos L.,University of Cádiz
Aquaculture Research | Year: 2013

The meagre (Argyrosomus regius) is a commercially important species for fisheries, and aquaculture production has been increasing in recent years. However, this growing importance has not been matched by increased genetic knowledge of the species. For this reason, an initial approach has been made to understanding the genetics of the meagre, using three particular multigene families (45S and 5S ribosomal DNA and U2 snRNA gene) as both molecular and cytogenetic markers. Only one type of 5S rDNA and only one ITS-1 (from 45S rDNA) were obtained, and these therefore could provide the basis for the development of genetic markers for this species. However, the U2 snRNA gene produced a multi-band electrophoretic pattern, and some of these bands were demonstrated to be linked with the U1 snRNA gene. This linkage could also provide a good species-specific marker which could be useful for tracing the evolutionary history of the Sciaenidae family. In this study, we have described for the first time the karyotype of the meagre: this is composed of 48 acrocentric chromosomes. Each of the three gene families were localized on different chromosome pairs, although the U2 snRNA gene probe was also located scattered throughout the genome of the meagre, as expected by the amplification pattern. © 2012 Blackwell Publishing Ltd.

Pawlus A.D.,University of Bordeaux 1 | Pawlus A.D.,University of Minnesota | Cantos-Villar E.,University of Bordeaux 1 | Cantos-Villar E.,Centro IFAPA | And 9 more authors.
Journal of Chromatography A | Year: 2013

Wine is a major dietary source of numerous potentially health promoting stilbenoids that have been the subject of many qualitative and quantitative studies. However, our initial HPLC-MS analyses of crude wine samples demonstrated the presence of compounds with molecular weights matching characteristic stilbenoid dimers, trimers, and tetramers that were unaccounted for in the literature. Due to the likelihood that these are known compounds, a chemical dereplication method is highly desirable. We developed such a method using LC-DAD-MS monitored fractionation steps, using adsorption and centrifugal partition chromatography (CPC), to obtain fractions rich in stilbenoids for analysis in stopped-flow LC-NMR. 1H NMR spectra and MS data were cross-referenced with our laboratory database and the literature for identification. This method yielded highly useful structural information, allowing the characterization of previously unidentified stilbenoids in wine, ampelopsin C, isohopeaphenol, quadrangularin A, and E-ω-viniferin. These results demonstrate the usefulness of stop-flow LC-NMR in conjunction with LC-MS guided fractionation for the dereplication of compounds of interest in general, and specifically for expanding the current knowledge of wine chemistry. © 2013 Elsevier B.V.

Domingues P.,Centro IFAPA | Garcia S.,Centro IFAPA | Hachero-Cruzado I.,Centro IFAPA | Lopez N.,National Autonomous University of Mexico | Rosas C.,National Autonomous University of Mexico
Aquaculture International | Year: 2010

The effects of two alternative prey (crayfish and hake) were tested on growth and survival of both juveniles and adults of Octopus vulgaris in two experiments. Octopuses fed the control (squid) were larger (3. 0 ± 0. 7 kg) than those fed crayfish (2. 4 ± 0. 6 kg) at the end of experiment I. Similarly, overall growth rates were higher for octopuses fed squid (1. 7 ± 0. 3 and 1. 2 ± 0. 2 %BW day-1, respectively). Average feeding rates for the experiment were not different, being 6. 5 ± 0. 9 and 7. 5 ± 0. 9 %BW day-1, respectively, for octopuses fed either squid or crayfish. Nevertheless, food conversions for the experiment were higher (42. 4 ± 2. 7%) for octopuses fed squid compared to the ones fed crayfish (23. 9 ± 1. 9 g). For experiment II, hake and crayfish were compared to squid; the final weight of octopuses fed squid, hake or crayfish was 1,183. 0 ± 242. 7 g, 1,175. 6 ± 240. 1 g and 922. 3 ± 160. 1 g, respectively. Overall growth rates for the experiment were 1. 9 ± 0. 2 %BW day-1, 1. 9 ± 0. 3 %BW day-1 and 1. 1 ± 0. 3 g, respectively. Final weight and growth rates were never different (P > 0. 05) between octopuses fed squid and hake, but were always higher (P < 0. 05) compared to the ones fed crayfish. Average feeding rates for experiment II were similar for the three diets, and of 4. 6 ± 1. 5, 4. 2 ± 1. 3 %BW day-1 and 5. 1 ± 0. 9 %BW day-1, respectively, for octopuses fed squid, hake or crayfish. Food conversions for experiment II were of 41. 0 ± 9. 6%, 40. 5 ± 9. 9% and 21. 3 ± 7. 4 g, respectively, for octopuses fed squid, hake or crayfish, and were always higher for octopuses fed squid and hake compared to crayfish. The results indicate that crayfish is not an adequate replacement for the usual prey to fatten octopus, even considering its much lower market price. © Springer Science+Business Media B.V. 2009.

Gutierrez A.,Instituto Valenciano Of Investigaciones Agrarias Ivia | Ruiz V.,Instituto Valenciano Of Investigaciones Agrarias Ivia | Molto E.,Instituto Valenciano Of Investigaciones Agrarias Ivia | Tapia G.,Centro IFAPA | del Mar Tellez M.,Centro IFAPA
Crop Protection | Year: 2010

The Red Palm Weevil (Rhynchophorus ferrugineus Olivier) is one of the worst pests for palms, since it can completely destroy the plant. Its early detection is very difficult and when the action of the pest is discovered, normally it is too late for recovering the plant. Bioacoustic sensors provide a good alternative for early detection of this pest because larvae activity is audible while they excavate tunnels to feed. This work describes the different stages for the development of an original electronic device based on acoustic sensors for early detection of Red Palm Weevil larvae in the interior of palms. This device is based on characteristic frequencies possibly related to their feeding activity. The performed tests have demonstrated that is possible to detect two week old larvae activity studying the sound intensity around 2250 Hz in palms infested only with 5 individuals under controlled environmental conditions. The activity of other insects artificially situated in the interior of palms did not affect the response of the device, possibly because they produce different frequencies (in the range 5-7 kHz). Future work will be aimed at assessing the infestation levels. © 2010 Elsevier Ltd.

Garcia-Garrido S.,Centro IFAPA | Hachero-Cruzado I.,Centro IFAPA | Garrido D.,Centro IFAPA | Rosas C.,National Autonomous University of Mexico | Domingues P.,Spanish Institute of Oceanography
Aquaculture International | Year: 2010

Lipid composition of the mantle and digestive gland of Octopus vulgaris that were not fed for 27 days were determined. Every 3 days, three octopuses were killed and samples of the mantle and the digestive gland (DG) were taken, in order to determine total lipids as well as lipid classes and fatty acids. Composition in total lipids (TL) for the mantle was similar until day 21, then decreased and remained similar until the end of the experiment. Composition in total lipids for the DG decreased significantly after 3 days, then remained similar until day 21, and then decreased until the end of the experiment. As for the lipid classes, in the DG the main components were triglycerides and sterol esters. Sterol esters suffered strong reductions after 10 days of starvation, while triglycerides remained similar until day 21 and then decreased until the end of the experiment. Cholesterol decreased gradually throughout the experimental period. For polar lipids, phosphatidylcholine and phosphatidylethanolamine increased during the first 3 days and then decreased throughout the experiment. In the mantle, the only neutral classes that decrease were triacylglycerols and sterol esters, while no polar lipid classes decreased in this organ. It was noticeable the decrease in almost all fatty acids in the DG after 3 days of starvation, while in the mantle there were no differences in fatty acid concentrations during the experiment. © 2010 Springer Science+Business Media B.V.

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