Ploidic and Molecular Analysis of ‘Morado de Huetor’ Asparagus (Asparagus officinale L.) Population; A Spanish Tetraploid Landrace

A study has been conducted into the genetic variation analysis of the tetraploid asparagus landrace ‘Morado de Huetor’ and its relationship with current commercial cultivars using Random Amplified Polymorphic DNA (RAPD) molecular markers. The presence of different ploidic levels in the ‘Morado de Huetor’ landrace has been also studied using cytogenetic analysis. Ten decamer oligonucleotides were used to obtain RAPD markers and to characterise 52 individuals of ‘Morado de Huetor’, 7 of the tetraploid cultivar ‘Purple Passion’, and 55 of 5 diploid hybrid commercial cultivars. Jaccard similarity index was calculated and a cluster analysis using UPGMA (Unweighted Pair-Group Method using Arithmetic Average) was performed. Tetraploid asparagus, ‘Morado de Huetor’ and ‘Purple Passion’, were well differentiated from the diploid ones showing a higher genetic variability. This result suggests a different origin for tetraploid and diploid asparagus varieties. Within the diploid cultivars a variety distribution was obtained. A specific monomorphic band (OPB20₈₈₃) was found in tetraploid varieties. Within the diploid cultivars, two bands (OPB20₈₃₀, OPC15₇₀₅) can be used to distinguish between the hybrid cultivars used in this study. Tri-, penta-, hexa-, and octoploid plants were found in ‘Morado de Huetor’. The origin of these ploidic levels is discussed and their use in the development of new varieties is proposed. In conclusion, ‘Morado de Huetor’ is a genetic resource that could be used to increase the narrow genetic background reported in diploid asparagus cultivars.     

Grain yield response of Zea mays (hybrid AE 703) to Azotobacter chroococcum H23

Summary Field experiments were conducted during 1985 and 1986 to examine the effect of Azotobacter chroococcum on the grain yield of maize. Application of 40 kg N ha^–1 plus A. chroococcum caused a significant increase in maize yield. Azotobacter inoculation was more efficient at lower doses (40 kg N ha^–1) than at high doses (80 kg N ha^–1) of urea.     

Validation of a monoclonal enzyme immunoassay for the determination of carbofuran in fruits and vegetables

The N-methylcarbamate pesticide carbofuran is a very important insecticide used worldwide. In the present work, the validation of a monoclonal antibody-based enzyme immunoassay (ELISA) to determine this compound in fruits and vegetables is described. The immunoassay is a competitive heterologous ELISA in the antibody-coated format, with an I(50) value for standards in buffer of 740 ng/L and with a dynamic range between 200 and 3100 ng/L. For recovery studies, peppers, cucumbers, strawberries, tomatoes, potatoes, oranges, and apples were spiked with carbofuran at 10, 50, and 200 ppb. After liquid extraction, analyses were performed by ELISA on extracts purified on solid-phase extraction (SPE) columns and crude, nonpurified extracts. Depending on the crop, mean recoveries in the 43.9--90.7% range were obtained for purified samples and in the 90.1--121.6% range for crude extracts. The carbofuran immunoassay performance was further validated with respect to high-performance liquid chromatography (HPLC) with postcolumn derivatization and fluorescence detection (EPA Method 531.1). Samples were spiked with carbofuran at several concentrations and analyzed as blind samples by ELISA and HPLC after SPE cleanup. The correlation between methods was very good (y = 0.90x + 2.66, r(2)() = 0.958, n = 25), with HPLC being more precise than ELISA (mean coefficients of variation of 4.1 and 11.5%, respectively). The immunoassay was then applied to the analysis of nonpurified extracts of the same samples. Results also compared very well with those obtained by HPLC on purified samples (y = 1.02x + 10.44, r(2)() = 0.933, n = 29). Therefore, the developed immunoassay is a suitable method for the quantitative and reliable determination of carbofuran in fruits and vegetables even without sample cleanup, which saves time and money and considerably increases the sample throughput.     

Comparison of the evolution of low molecular weight phenolic compounds in typical Sherry wines: Fino,Amontillado, and Oloroso

Changes in the content of low molecular weight phenolic compounds (hydroxybenzoic and hydroxycinnamic acids, aldehydes, and their esterified derivatives, tyrosol and 5-(hydroxymethyl)-2-furaldehyde) during the aging of three different classes of Sherry wine, Fino, Oloroso, and Amontillado, have been studied. The samples studied were taken from each of the scales of the particular aging system applied to the three classes of wine. Clear differences were observed in the behavior of the low molecular weight phenolic in the three classes of wine. The wines subjected to oxidative aging presented a higher phenolic content overall, with the exception of the esterified derivatives of phenolic compounds that are mainly found in the samples that have not undergone any process of oxidation. MANOVA results confirmed that there are significant differences between all of the samples of the three types of wines. Using LDA, a classification of 100% of the samples has been made.     

Effect of gluconic acid consumption during simulation of biological aging of sherry wines by a flor yeast strain on the final volatile compounds

Flor yeast Saccharomyces cerevisiae (capensis G1) strain assimilates gluconic acid during the aerobic biological aging process of sherry wines and exerts significant changes on the final volatile compounds of wines, especially a decrease in volatile acidity and butanoic, isobutanoic, 2-methylbutanoic, and 3-methylbutanoic acids. This decrease may have a favorable effect on the quality of sherry wines.     

Germplasm Management of Faba Bean (Vicia faba L.): Monitoring Intercrossing between Accessions with Inter-plot Barriers

Knowledge of gene flow is essential for designing strategies of germplasm multiplication. Inter and intra-plot gene flow and pollen dispersion between small plots in a field of Vicia faba germplasm multiplication were measured under four isolation zones at two locations. The four isolation zones were: a barren zone, a nucleocytoplasmic Vicia faba male sterile line, a Vicia faba tetraploid genotype and a Vicia narbonensis L. population. Genotypes fixed for alternative isozyme and allozyme alleles allowed the identification of inter and intra-plot hybrids through progeny testing. A set of log-likelihood ratio (LLR) G tests were used to analyze the effects of geographical location, isolation zone and genotype on the different components of gene flow. Considerable heterogeneity among the genotypes in all the components, inter and intra-plot gene flow and pollen dispersion, of gene flow has been found. It has been shown that gene flow is location, isolation zone and genotype dependent. Planting a barrier surrounding the plots rather than using a non cultivated area between plots seemed more or less efficient for preventing inter-plot gene flow depending on the genotype and the location. Specific associations between genotype and male sterile or tetraploid border rows for every location can bring about gene flow as high, or even higher, than those produced by a barren zone. Differences among the genotypes and borders in flowering overlapping appear not to be the major factor influencing the pattern of gene flow. Implications of the pattern of gene flow observed are discussed in relation to crop diversity managed by farmers.     

Evaluation of the cytotoxicity, genotoxicity, mutagenicity, and antimutagenicity of propolis from Tucuman, Argentina

This study evaluates the toxic, genotoxic/mutagenic, and antimutagenic effects of propolis extract from Amaicha del Valle, Tucumán, Argentina. The cytotoxicity assays carried out with the lethality test of Artemia salina revealed that the LD50 was around 100 microg/mL. Propolis extracts showed no toxicity to Salmonella typhimurium TA98 and TA100 strains and Allium cepa at concentrations that have antibiotic and antioxidant activities. Otherwise, for the testing doses, neither genotoxicity nor mutagenicity was found in any sample. The propolis extracts were able to inhibit the mutagenesis of isoquinoline (IQ) and 4-nitro o-phenylenediamine (NPD) with ID50 values of 40 and 20 microg/plate, respectively. From this result, the studied propolis may be inferred to contain some chemical compounds capable of inhibiting the mutagenicity of direct-acting and indirect-acting mutagens. A compound isolated from Amaicha del Valle propolis, 2',4'-dihydroxychalcone, showed cytotoxic activity (LC50 values of 0.5 microg/mL) but was not genotoxic or mutagenic. Furthermore, this compound was able to inhibit the mutagenicity of IQ (ID50 values of 1 microg/plate) but was unable to inhibit the mutagenicity of NPD. Our results suggest a potential anticarcinogenic activity of Amaicha del Valle propolis and the chalcone isolated from it.     

Genotype and harvest time influence the phytochemical quality of Fino lemon juice (Citrus limon (L.) Burm. F.) for industrial use

Two clonal selections of lemon tree (Citrus limon Burm. f. cv. Fino), named Fino-49-5 and Fino-95, were studied to ascertain the influence of genetic (clone) and environmental (season) factors on the human-health bioactive compounds of lemon juice (vitamin C and flavonoids) and the possible relationship between composition and in vitro antioxidant capacity (2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid), and ferric reducing antioxidant power) of the juice. The cultivar Fino-49-5 performed better in terms of flavonoid and vitamin C contents. Variability in the weather conditions determined, at least in part, differences in the content of lemon juice bioactives more importantly than the genetic background did. Therefore, the food industry would have phytochemically rich and nutritive lemons with practically complete independence of the harvest time and the selected cultivar.     

Natural and Anthropogenic Contributions to PM10 and PM2.5 in an Urban Area in the Western Mediterranean Coast

Source apportionment analysis was used to identify the factors contributing to atmospheric pollution at a monitoring location in the Southeast of Spain, a well documented area with an arid climate and high insolation favouring two sources of particulate matter: secondary transformation in the atmosphere and resuspension of crustal dry soils to the air. These conditions are further complicated by numerous industrial facilities in the area of the historical city of Cartagena. This paper describes the air quality of an area which includes a zinc metallurgical industry, a petrochemical factory, an oil power station, a shipyard and natural phenomena including African dust transport and resuspension of regional and/or local crustal materials. Major and trace element concentrations in PM10 and PM2.5 were determined at two monitoring stations in Cartagena (one PM10 sampler located at a traffic hotspot and the PM2.5 sampler at a suburban station), during 2004 and 2005. Results showed that in the PM10 fraction, the zinc metallurgical activity was linked to high levels of Cd, Zn and Pb; shipyard emission was associated with high levels of Cr and Ni; and high Ni and V levels were associated with the secondary aerosol indicating the contribution from oil combustion (oil-fired power station or petrochemical facilities). In the PM2.5 size fraction, the zinc source is defined by Zn and Pb; V, Ni and As appear with the oil combustion emissions. In contrast to PM10, shipyard activity is not consistently defined. Consistent sources found in both size fractions include crustal materials and traffic emissions.     

Molecular and antimicrobial susceptibility characterization of Globicatella sulfidifaciens isolated from sow's urinary tract infection

Background: The Globicatella genus comprises Gram-positive, facultative anaerobic, α-hemolytic and catalase negative cocci morphologically and phenotypically very similar to Streptococcus and Aerococcus genus which can lead to misidentification and underestimation of this pathogen. Globicatella species have already been isolated from human and animals with heart and brain disorders. Their clinical relevance in animals, and its zoonotic potential, remains unknown due to the difficulty in their identification.

Objective: To present the isolation, phenotypic and molecular characterization of G. sulfidifaciens from urinary tract infection in sows.

Materials and Methods: Urine samples from 140 sows of two swine herds located in São Paulo State (Brazil) yielded the isolation of three presumptive G. sulfidifaciens strains. Identification and species confirmation were done by MALDI-TOF MS and 16S rRNA sequencing. Strains were further characterized by single enzyme amplified fragments length polymorphism (SE-AFLP) and broth microdilution techniques.

Results: All three isolates were confirmed as G. sulfidifaciens. The SE-AFLP genotyping resulted in distinct fingerprint patterns for each strain. All isolates presented high MIC values to tetracycline, sulphonamides, aminoglycosides and tylosin tartrate, which present high usage in human and animal medicine.

Conclusions: Globicatella sulfidifaciens could be related to sporadic urinary tract infections in swine and appear to present alarming antimicrobial susceptibility profile. It is necessary to differentiate Streptococcus-like microorganisms in routine laboratory diagnostics for the correct identification of underestimated species potentially pathogenic to animals.