ITS1-rDNA-based methodology to identify world-wide hake species of the Genus Merluccius

Species-specific DNA-based tags are valuable tools for the management of both fisheries and commercial fish products. In this study, we have developed a two-step molecular tool to detect the presence of hake DNA (Merluccius spp.) and to identify the exact hake species present in an blind sample. The first test involves PCR amplification of an ITS1-rDNA fragment of 193 bp using nested primers that are interspecifically conserved in Merluccius spp. and Atlantic cod, Gadus morhua. The second test consists of the PCR amplification of a 602-659 bp DNA fragment spanning part of the ribosomal cluster 18S-ITS1-5.8S and digesting it with four restriction enzymes whose targets map at interspecifically nonconserved sites of the ITS1. Alternatively, the identification of hake species can be achieved by FINS or BLAST, using the nucleotide sequence of either the whole ITS1 sequence or its nested fragment of 193 bp. Because of their high reproducibility and ease of execution, these procedures allow for routine analysis and constitute high reliable tools for the rapid identification of 12 species of hake.     

Combined use of gliadins and SSRs to analyse the genetic variability of the Spanish collection of cultivated diploid wheat (Triticum monococcum L. ssp. monococcum)

This work studied the combined use of gliadins and SSRs to analyse inter- and intra-accession variability of the Spanish collection of cultivated einkorn (Triticum monococcum L. ssp. monococcum) maintained at the CRF-INIA. In general, gliadin loci presented higher discrimination power than SSRs, reflecting the high variability of the gliadins. The loci on chromosome 6A were the most polymorphic with similar PIC values for both marker systems, showing that these markers are very useful for genetic variability studies in wheat. The gliadin results indicated that the Spanish einkorn collection possessed high genetic diversity, being the differentiation large between varieties and small within them. Some associations between gliadin alleles and geographical and agro-morphological data were found. Agro-morphological relations were also observed in the clusters of the SSRs dendrogram. A high concordance was found between gliadins and SSRs for genotype identification. In addition, both systems provide complementary information to resolve the different cases of intra-accession variability not detected at the agro-morphological level, and to identify separately all the genotypes analysed. The combined use of both genetic markers is an excellent tool for genetic resource evaluation in addition to agro-morphological evaluation.     

Interactive effect between Cu‐adapted arbuscular mycorrhizal fungi and biotreated agrowaste residue to improve the nutritional status of Oenothera picensis growing in Cu‐polluted soils

The interactive effect of sugar beet (SB) agrowaste and arbuscular mycorrhizal (AM) fungi inoculation in response to increasing Cu levels was evaluated in the metallophyte Oenothera picensis. Plants were grown in a Cu‐added soil (0, 100, or 500 mg Cu kg^?1), in presence or absence of SB, and inoculated with: (1) indigenous Cu adapted mycorrhiza (IM) isolated from Cu‐polluted soils; (2) Claroideoglomus claroideum (CC); or (3) maintained uninoculated (control). Sugar beet application produced an increase in shoot biomass of 2 to 7 times, improving plant nutritional status and allowing their survival at the highest Cu concentrations. Moreover, AM fungi utilization had a positive effect promoting the plant establishment; nevertheless, Cu plant concentration as well as the mycorrhizal development in terms of AM colonization, AM spore density, and glomalin production were strictly dependent of the AM fungi strains used. Remarkable differences between AM fungi strains were observed at the highest soil Cu level where only plants colonized by IM were able to survive and grow when no SB residue was added. An interactive effect between AM fungi and SB produced a higher plant growth than plants without the amendment application, improving the plant establishment and allowing their survival at highest copper concentrations, suggesting that this combination could be used as a biotechnological tool for the phytoremediation of Cu‐polluted soils.     

Influence of fruit ripeness at the time of seed extraction on pepper (Capsicum annuum) seed germination

The influence of fruit ripeness (half ripe, fully ripe, overripe) at the time of seed extraction on seed germination behaviour, at 25 °C and 13 °C, was studied in two Spanish pepper cultivars for canning. Seeds from half ripe fruits had a poorer behaviour than those taken from fully ripe fruits, especially at 13 °C. Room ripening and overripening of half ripe fruits improved the germination behaviour of the seeds although this behaviour was poorer than that of seeds extracted from fully ripe fruits that were allowed to overripen. Overripening of fully ripe picked fruits slightly improved germination behaviour.     

Crossability relationships between the common potato, Solanum tuberosum spp. tuberosum, and its wild diploid relatives S. kurtzianum and S. ruiz-lealii

The common potato, Solanum tuberosum spp. tuberosum (tbr), is a tetraploid species with a narrow genetic base, but with a large number of related species that harbor great genetic diversity for numerous characters of agronomic interest. S. kurtzianum (ktz) and S. ruiz-lealii (rzl) are wild diploid relatives with potential resistance to adverse biotic and abiotic factors. To evaluate if this wild germplasm can be incorporated by conventional crosses into the tbr gene pool, pollen-pistil compatibility relations and seed set in interspecific crosses with tbr were investigated. In 4x tbr × 2x ktz crosses and the reciprocals, 89% and 52.2% of genotypic combinations (respectively) were compatible at the pollen-pistil level. Seeds were obtained from some genotypic combinations in both directions of the cross, suggesting that functional 2n gametes might be produced by particular ktz genotypes. In 2x rzl × 4x tbr crosses and their reciprocals, 35% and 11.7% (respectively) of the genotypic combinations were compatible at the pollen-pistil level, but no seeds were obtained. These results indicate that the reproductive isolation between 4x tbr and 2x ktz is incomplete, and that gene exchange between them is feasible. Moreover, further studies will have to be carried out with rzl to ascertain its crossability with 4x tbr.     

Avascular necrosis of the femoral heads in a red panda (Ailurus fulgens fulgens): possible Legg-Calve-Perthes disease.

A 17-mo-old captive-born female red panda (Ailurus fulgens fulgens) presented with a sudden onset of lameness in its left hind leg was diagnosed radiographically as having possible severe, bilateral Legg-Calve-Perthes disease with fracture of the great trochanter of the left femur. Surgical repair of the fracture was performed using pins and a tension band wire through a lateral approach to the hip. This is the first case reported at Madrid Zoo-Aquarium, where 63 individuals have been bred over 15 yr.     

Can place names be used as indicators of landscape changes? Application to the Doñana Natural Park (Spain)

This work broaches the possibility of using place names as indicators of original landscapes that have been much transformed. The reconstruction of landscape elements from place names is commonly disputed because such daring notion is impossible to demonstrate. The present case avoids this by making a preliminary study of changes in the landscape using conventional methods. With the knowledge gained from objective and reliable sources, the possibility is analyzed of whether place names are a reflection of landscape changes taking place over a considerable period of time (the last few centuries). It is concluded that, for the present case study, in natural areas with a high rate of change of land use (Doñana Natural Park), place names indicate not only changes in the landscape, but also how such changes are perceived. In the study area, this is especially clear regarding the fens.     

In vitro micrografting and its applications to fruit science

Improvements have been brought to in vitro micrografting. The breaking of dormancy by dipping the seeds of the rootstock in concentrated solutions of benzylaminopurine avoids having recourse to long stratification in the cold (60–120 days according to species). The initial pre-treatment of apices with cytokinin allows their development and facilitates later grafting. Moreover, adding DIECA to the cut sections avoids oxidation and, together with the insertion of a piece of agar between apex and rootstock, increases the success of micrografting. Finally, a special technique of in vivo micrografting of pretreated apices on young plants has been developed.Micrografting is commonly used to obtain peach trees free of Sharka and NRS viruses, and to study incompatibility mechanisms in heterografts, leading to an early diagnosis of incompatibilities.     

Pathophysiology of hepatic coccidiosis in rabbits

Four groups of 5 40-day-old, Eimeria stiedai-naive, New Zealand rabbits were infected with 0 (Group A), 10² (B), 10³ (C) and 10⁴ (D) sporulated oocysts of E. stiedai and observed for 50 days. Serum glutamic pyruvic (GPT) and glutamic oxalacetic (GOT) transaminases, bilirubinemia, lipemia, proteinemia, glycemia, oocyst output, body, carcass and liver weights, and mortality were recorded. Four physiopathological events were identified: (1) a phase of indirect damage to the hepatocytes that takes place during the first 2 weeks of infection and is characterized by increased transaminases; (2) a cholestatic period consequent to the production of oocysts that begins suddenly in the 3rd week, diminishes gradually towards the 7th week, and is characterized by a rise of bilirubinemia and lipemia; (3) a stage of metabolic dysfunction that begins in the 3rd–4th week, intensifies for the next 3 weeks and starts to recover during the 7th week. It is characterized by hypoproteinemia and hypoglycemia; (4) a period of immunodepression characterized by the inability of the heavily infected host to inhibit oocyst production. Apart from the cholestatic phase, the respective pathogenic mechanisms remains to be studied.