Genetic characterization of a collection of chayote, <Emphasis Type="Italic">Sechium edule</Emphasis> (Jacq.) Swartz,in Costa Rica by using isozyme markers

We established protocols for the analysis of genetic diversity in chayote (Sechium edule) by using isozyme markers, thereby determining the level of genetic diversity present in 42 accessions of chayote from Costa Rica. We obtained clear and reproducible zymograms for eight enzyme staining systems: PGM, 6-PGD, PGI, IDH, MDH, SOD, SKD, and EST, and were able to score 14 putative loci. Eight of the 14 loci examined were polymorphic. We found 35 distinct multilocus genotypes among these accessions. Five of these multilocus genotypes were homozygous for all loci. In addition, our data also revealed that most of the multilocus genotypes (24) were heterozygous for only one of the eight loci, and the rest were heterozygous for two or three loci (9 and 4 accessions, respectively). Seven multilocus genotypes were found in two different accessions. Dice similarity coefficient was used to study the relationship between accessions. This analysis, based on the presence and absence of alleles, revealed that accessions collected in the same location seldom shared the same multilocus genotype. The value of isozyme polymorphisms as tools to continue studies on the characterization of chayote is discussed.     

Isozyme diversity and analysis of the mating system of the wild rice Oryza latifolia Desv. in Costa Rica

Oryza latifolia is the most abundant and widespread wild relative of rice in the lowlands of Costa Rica. It also has the most diverse morphology. However, little is known about the genetic diversity and mating system of this allotetraploid species. Genetic analyses of nine populations from different life zones from the Pacific and Atlantic slopes were performed using six isozyme loci. Differences in genetic variability were observed among populations, of which Palo Verde and Santa Rosa were the most diverse. Most Atlantic populations clustered together, and a similar result was obtained with the Pacific populations. High levels of interpopulation diversity were observed while most populations were monomorphic for at least one genotype, suggesting little genetic flow within populations. Even in polymorphic populations no variation was observed within progeny groups, combined with Hardy-Weinberg disequilibria in most populations, commonly observed in autogamous species or in species with clonal reproduction. However, the high frequency of heterozygous-like patterns may suggest that the reproductive system of O. latifolia might be more complex. Future research may explain the genetic patterns as well as the reproductive biology of this species. The knowledge of the genetic diversity and mating system of O. latifolia could contribute to the implementation of genetic resource conservation strategies and gene flow analyses, as well as of breeding programs for rice improvement.     

Mate choice of an endogeic earthworm revealed by microsatellite markers

Endogeic earthworms are difficult study subjects due to the cryptic medium in which they live; thus, only the behaviour of epigeic and anecic earthworms has been studied before. We used microsatellite markers as a tool to elucidate the mate choice processes of Hormogaster elisae, an endogeic earthworm. It was shown to normally mate with two partners, preferably of the same size that are found in close proximity thereby eliminating the need for long-distance dispersion, which could explain the previously observed high genetic differentiation between populations. The genetic analyses of the sperm within each of its four spermathecae showed a uniform distribution with no signs of differential storage of sperm from different partners.     

Chemical characterization of lignin and lipid fractions in industrial hemp bast fibers used for manufacturing high-quality paper pulps

The chemical composition of lignin and lipids of bast fibers from industrial hemp (Cannabis sativa) used for high-quality paper pulp production was studied. Pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) of fibers showed a lignin with a p-hydroxyphenyl:guaiacyl:syringyl unit (H:G:S) molar proportion of 13:53:34 (S/G ratio of 0.64). p-Hydroxycinnamic acids, namely, p-coumaric and ferulic acids, were found in only trace amounts. Among the lipids, the main compounds identified by GC/MS of the hemp fibers extracts were series of n-alkanes, free and esterified sterols and triterpenols, waxes, and long-chain n-fatty acids. Other compounds such as n-aldehydes, n-fatty alcohols, steroid hydrocarbons, and steroid and triterpenoid ketones as well as steryl glycosides were also found.     

Tartaric acid recovery from distilled lees and use of the residual solid as an economic nutrient for lactobacillus

The recovery of tartaric acid (TA) from distilled vinification lees coming from the white and red winemaking technology was optimized using response surface methodology and Statistica 5.0 software. The sequential treatment of dissolving TA and further calcium tartrate (CaT) precipitation could be used to recover up to 92.4% of the initial TA when distilled white lees were used. The residual lees were employed as economic nutrients for lactic acid production by Lactobacillus pentosus CECT-4023 using hemicellulosic vine shoot hydrolysates as carbon source. Distilled lees after TA extraction used as nutrients provided values of lactic acid (18.4-18.9 g/L), global volumetric productivities (0.82-0.84 g/L.h), and product yields (0.69-0.70 g/g) similar to those achieved when using the general medium for Lactobacilli (18.6 g/L, 1.11 g/L.h, and 0.62 g/g, respectively) or lees without TA extraction (16.4-17.2 g/L, 0.96-1.21 g/L.h, and 0.61-0.66 g/g, respectively). This technology not only avoids pollutant disposal but also represents a commercial source of tartaric acid and economic nutrients for biotechnological processes.     

Bioactive cinchona alkaloids from Remijia peruviana

Three known Cinchona alkaloids of the quinine type, quinine (1), cupreine (2), cinchonine (3), and the possible artifact cinchonine-HCl (3-HCl), along with two new ones, acetylcupreine (4) and N-ethylquinine (5), have been isolated from the bark of Remijia peruviana (Rubiaceae). Their stereochemical structures were established by high resolution NMR spectroscopy. Alkaloids 2-4 had antifeedant effects on Leptinotarsa decemlineata with varying potencies. Compound 4 was cytotoxic to both insect Sf9 and mammalian CHO cells after 48 h of incubation, while 3-HCl had stronger and selective cytotoxicity to Sf9. Quinine 1 had a moderate to low effect on Trypanosoma cruzi. Tumoral cells were also affected by these alkaloids, with 4 and 3-HCl being the most cytotoxic to all the cell lines tested. Overall, the 8R, 9S configurations, as in 3 and 3-HCl, as well as the C-6'acetylated alkaloid 4, with an 8S, 9R configuration, showed stronger biological effects.     

Effect of high-intensity pulsed electric fields processing and conventional heat treatment on orange-carrot juice carotenoids

Liquid chromatography (LC) was the method of choice for quantification of carotenoids (including geometrical isomers) to evaluate the effects of high-intensity pulsed electric field (HIPEF), a nonthermal preservation method, with different parameters (electric field intensities and treatment times), on an orange-carrot juice mixture (80:20, v/v). In parallel, a conventional heat treatment (98 degrees C, 21 s) was applied to the juice. HIPEF processing generally caused a significant increase in the concentrations of the carotenoids identified as treatment time increased. HIPEF treatment at 25 and 30 kV/cm provided a vitamin A concentration higher than that found in the pasteurized juice.     

Detection of ruminant meat and bone meals in animal feed by real-time polymerase chain reaction: result of an interlaboratory study

The commercialization of animal feeds infected by prions proved to be the main cause of transmission of bovine spongiform encephalopathy (BSE). Therefore, feed bans were enforced, initially for ruminant feeds, and later for all feeds for farmed animals. The development and validation of analytical methods for the species-specific detection of animal proteins in animal feed has been indicated in the TSE (Transmissible Spongiform Encephalopathies) Roadmap (European Commission. The TSE (Transmissible Spongiform Encephalopathy) roadmap. URL: http://europa.eu.int/comm/food/food/biosafety/bse/roadmap_en.pdf, 2005) as the main condition for lifting the extended feed ban. Methods based on polymerase chain reaction (PCR) seem to be a promising solution for this aim. The main objective of this study was to determine the applicability of four different real-time PCR methods, developed by three National expert laboratories from the European Union (EU), for the detection and identification of cattle or ruminant species in typical compound feeds, fortified with meat and bone meals (MBM) from different animal species at different concentration levels. The MBM samples utilized in this study have been treated using the sterilization condition mandatory within the European Union (steam pressure sterilization at 133 degrees C, 3 bar, and 20 min), which is an additional challenge to the PCR methods evaluated in this study. The results indicate that the three labs applying their PCR methods were able to detect 0.1% of cattle MBM, either alone or in mixtures with different materials such as fishmeal, which demonstrates the improvement made by this technique, especially when compared with results from former interlaboratory studies.