Determination of organic acids in tissues and exudates of maize, lupin, and chickpea by high-performance liquid chromatography-tandem mass spectrometry

This article describes a fast and simple methodology for the extraction and determination of organic acids in tissues and root exudates of maize, lupin, and chickpea by LC/MS/MS. Its main advantage is that it does not require sample prepurification before HPLC analysis or sample derivatization to improve sensibility. The results obtained showed good precision and accuracy, a recovery close to 100%, and no significant matrix effect. Moreover, the sensibility of the method is in general better than that of previously described methodologies, with detection limits between 15 and 900 pg injected.     

Fine-Scale Spatial Genetic Structure in Perennial Grasses in Three Environments

Past research has shown that changes in grazing-resistance traits may be associated with genetic changes in plant populations. Little is known about spatial genetic relationships within plant populations (spatial genetic structure) and any grazing effects on these relationships. Here we present observations of the fine-scale spatial genetic structure in three grass species in semiarid environments (Arizona, Mexico, and Argentina). In each environment, populations of a dominant grass species were sampled from two sites with contrasting livestock grazing histories. Plant genotypes were described with the use of amplified fragment length polymorphism markers. In Arizona, populations of sideoats grama (Bouteloua curtipendula var. caespitosa Gould and Kapadia) differed in that one has never experienced livestock grazing, whereas cattle have grazed the other. In the other two environments, populations exposed to long-term heavy grazing were examined, along with those that experienced either moderate grazing (Mexico, blue grama [Bouteloua gracilis {Willd. ex Kunt} Lag. ex Griffiths]) or extended exclusion of livestock (Argentina [Poa ligularis Nees ex Steud.]). Based on independent analysis of each population, we observed no differences in average gene diversity between populations of each species. With the use of analysis of molecular variance we found slight but significant genetic differentiation between populations with different grazing histories in Arizona and Argentina. Significant genetic structure was present in all populations and indicated an inverse relationship between spatial and genetic distance. Interestingly, this relationship was most pronounced in the cattle-free sideoats grama population, suggesting larger genetic neighborhood areas in the absence of livestock. Less distinct differences in spatial genetic structure associated with grazing history were evident in the other two species. We hypothesize that livestock grazing may lead to increased homogeneity in genetic structure at the landscape scale. Effectively examining this hypothesis presents many experimental challenges.     

Results from an indirect fluorescent antibody test using three different strains of Ehrlichia canis

An indirect fluorescent antibody (IFA) test is usually performed to detect antibodies in dogs naturally infected by Ehrlichia canis. In this work, results obtained using three different E. canis strains as antigen (a commercial antigen, the E. canis Oklahoma strain and the E. canis Madrid strain) were compared. One hundred and forty-nine serum samples obtained from dogs living in the centre of Spain were analysed. When qualitative results were evaluated, identical results were detected in 87.2% of samples for the three antigens tested. When comparing antibody titre results, differences between the Madrid strain and the commercial antigen, and between the Madrid and Oklahoma strains were statistically significant (P < 0.0001). No differences were found when comparing the Oklahoma strain with the commercial antigen (P = 0.562). Subtle intra-laboratory variations shown in this study suggest a higher sensitivity of the IFA test when an autochthonous strain is used as antigen.     

Direct aqueous injection liquid chromatography/electrospray ionization-mass spectrometry/mass spectrometry analysis of water for atrazine, simazine, and their chlorotriazine metabolites

A method is reported for the determination of atrazine, simazine, and their respective dealkylated chlorotriazine metabolites in ground, surface, and finished drinking water. Water samples are diluted 1:4 in an injection vial prior to analysis using liquid chromatography/electrospray ionization-mass spectrometry/mass spectrometry (LC/ESI-MS/MS). The lower limit of method validation is 0.10 microg/L (ppb) for 2-chloro-4-(ethylamino)-6-isopropylamino)-s-triazine (atrazine, G-30027), 2-chloro-4, 6-(diethylamino)-s-triazine (simazine, G-27692), 2-amino-4-chloro-6-(isopropylamino)-s-triazine (deethylatrazine, DEA, or G-30033), 2-amino-4-chloro-6-(ethylamino)-s-triazine (deisopropylatrazine, DIA, or G-28279), and 2,4-diamino-6-chloro-s-triazine (didealkylatrazine, DDA, or G-28273). The overall mean procedural recoveries (and % relative standard deviations) for atrazine, simazine, DEA, DIA, and DDA are 98 (4.4), 102 (3.6), 99 (4.8), 103 (4.0), and 109% (4.8%), respectively, in finished drinking water; 108 (2.7), 104 (5.4), 113 (4.5), 111 (5.2), and 105% (5.3%), respectively, in groundwater; and 96 (6.9), 103 (4.2), 102 (4.4), 102 (5.2), and 102% (8.2%), respectively, in surface water. The method validation was conducted under U.S. EPA FIFRA Good Laboratory Practice Guidelines 40 CFR 160.     

Toxicity of cultured bullseye puffer fish Sphoeroides annulatus

The toxin content in various life cycle stages of tank-cultivated bullseye puffer (Sphoeroides annulatus) were analyzed by mouse bioassay and ESI-MS spectrometry analysis. The presence of toxin content was determined in extracts of sperm, eggs, embryo, larvae, post-larvae, juvenile, pre-adult, and adult fish, as well as in food items used during the cultivation of the species. Our findings show that only the muscle of juveniles, the viscera of pre-adults, and muscle, liver, and gonad of adult specimens were slightly toxic (<1 mouse unit). Thus, cultivated S. annulatus, as occurs with other cultivated puffer fish species, does not represent a food safety risk to consumers. This is the first report of toxin analysis covering the complete life stages of a puffer fish under controlled conditions.     

Phlorotannin Extracts from Fucales Characterized by HPLC-DAD-ESI-MSn: Approaches to Hyaluronidase Inhibitory Capacity and Antioxidant Properties

Purified phlorotannin extracts from four brown seaweeds (Cystoseira nodicaulis (Withering) M. Roberts, Cystoseira tamariscifolia (Hudson) Papenfuss, Cystoseira usneoides (Linnaeus) M. Roberts and Fucus spiralis Linnaeus), were characterized by HPLC-DAD-ESI-MSⁿ. Fucophloroethol, fucodiphloroethol, fucotriphloroethol, 7-phloroeckol, phlorofucofuroeckol and bieckol/dieckol were identified. The antioxidant activity and the hyaluronidase (HAase) inhibitory capacity exhibited by the extracts were also assessed. A correlation between the extracts activity and their chemical composition was established. F. spiralis, the species presenting higher molecular weight phlorotannins, generally displayed the strongest lipid peroxidation inhibitory activity (IC₅₀ = 2.32 mg/mL dry weight) and the strongest HAase inhibitory capacity (IC₅₀ = 0.73 mg/mL dry weight). As for superoxide radical scavenging, C. nodicaulis was the most efficient species (IC₅₀ = 0.93 mg/mL dry weight), followed by F. spiralis (IC₅₀ = 1.30 mg/mL dry weight). These results show that purified phlorotannin extracts have potent capabilities for preventing and slowing down the skin aging process, which is mainly associated with free radical damage and with the reduction of hyaluronic acid concentration, characteristic of the process.     

High Antioxidant Activity Mixture of Extruded Whole Quality Protein Maize and Common Bean Flours for Production of a Nutraceutical Beverage Elaborated with a Traditional Mexican Formulation

The objective of this study was to determine the best combination of extrusion process variables for the production of whole quality protein maize (EQPMF) and common bean (ECBF) flours to prepare a high antioxidant activity mixture (EQPMF + ECBF) suitable to produce a nutraceutical beverage with high acceptability elaborated with a traditional Mexican formulation. Processing conditions were obtained from a factorial combination of barrel temperature (BT?=?120–170 °C) and screw speed (SS?=?120–200 rpm). Response surface methodology was applied to obtain maximum values for antioxidant activity (A _ox A) of the flour mixture (EQPMF + ECBF) and acceptability (A) of the nutraceutical beverage. The best combinations of extrusion process variables for EQPMF and ECBF to prepare an optimized mixture (60%EQPMF?+?40%ECBF) were BT?=?98 °C/SS?=?218 rpm and BT?=?105 °C/SS?=?83 rpm, respectively. The optimized mixture had A _ox A?=?14,320 μmol Trolox equivalent (TE)/100 g sample dry weight (dw) and a calculated protein efficiency ratio (C-PER) of 2.17. A 200 ml portion of a beverage prepared with 25 g of the optimized flour mixture had A _ox A?=?3,222 μmol TE, and A?=?89 (level of satisfaction “I like it extremely”). This nutraceutical beverage could be used as an alternative to beverages with low nutritional/nutraceutical value, such as those prepared with water, simple sugars, artificial flavoring and colorants, which are widely offered in the market.     

Improved Microbial and Chemical Reduction of Direct Blue 71 Using Anthraquinone-2,6-disulfonate Immobilized on Granular Activated Carbon

The aim of this study was to evaluate the redox mediating capacity of anthraquinone-2,6-disulfonate (AQDS) immobilized on granular activated carbon (GAC) during the reductive decolorization of direct blue 71 (DB71) under microbial and chemical conditions. The immobilization of AQDS on GAC was conducted by adsorption, and it has obtained an uptake capacity of 0.227 mmol g^?1. The anchorage of AQDS on GAC improved its electron transfer capacity (ETC) up to 2.05 times higher than the raw material. Similarly, the addition of GAC-AQDS increased up to 1.75- and 1.16-fold the rate of decolorization (k _d ) of DB71 under microbial and chemical conditions, respectively, in comparison to the unmodified GAC. Surprisingly, a higher k _d value was achieved in incubations without either GAC or GAC-AQDS because of the generation of aromatic amines, from the reduction DB71, taking into account that these species may act as a catalyst in the DB71 reduction process. In contrast, adsorption of aromatic amines on either GAC or GAC-AQDS decreased its redox mediating capacity as evidenced by spectrophotometric screenings of the decolorized solution and the supporting material. The development of materials with enhanced both redox and adsorption properties, as the GAC used in this study, offers a promising way to increase the redox conversion of recalcitrant pollutants commonly found in industrial wastewaters.     

Nutraceutic effect of free condensed tannins of <Emphasis Type="Italic">Lysiloma acapulcensis</Emphasis> (Kunth) benth on parasite infection and performance of Pelibuey sheep

Forty-five Pelibuey sheep were experimentally infested with nematodes to evaluate the effect of three free condensed tannin (FCT) levels of Lysiloma acapulcensis on fecal egg counts (FECs), packed cell volumes (PCV), ocular mucosa colors (OMC), average daily gain (ADG), and adult nematode count. Five treatments were used: 12.5, 25.0, and 37.5 mg of FCT kg^?1 of body weight (BW); sterile water (control); and ivermectine (0.22 mg kg^?1 of BW) as chemical group. The data were processed through repeated measurement analysis. Even though the three FCT doses decreased (P?<?0.05) the FEC, the highest reduction was obtained with 37.5 mg kg^?1 of BW. No differences were observed in PCV and OMC. Higher ADG (P?<?0.05) was observed with 37.5 mg kg^?1 of BW of FCT. The count of adult nematodes (females and males) in the higher dose of FCT was similar to chemical treatment. Dose of 37.5 mg kg^?1 of BW decreased the parasite infection and improved the lamb performance. Therefore, this dose could be used as a nutraceutic product in sheep production.     

Grain composition of Virginia winter barley and implications for use in feed,food, and biofuels production

Grain compositional components impacting barley (Hordeum vulgare L.) use in food, feed and fuel products, must be combined with improved agronomic traits to produce a commercially viable barley cultivar. Little current information is available on grain composition and variability among winter barley genotypes. This study was conducted to determine the variability among modern hulled and hulless winter barley genotypes in grain composition. Barley types varied significantly in grain and kernel weight, starch, beta-glucan, oil and ash content, but not in protein concentration. Hulless barley had significantly higher grain test weight and starch concentration than hulled and malting types, and significantly higher beta-glucan than malting barley. Hulless barley had significantly lower kernel weights, oil, and ash concentrations than hulled and malting types. Higher starch and lower fiber and ash in grain of hulless barley versus hulled feed or malting type barley are characteristics that increase hulless barley desirability. Selection for high starch concentration among all barley types is feasible and will facilitate development of barley cultivars better suited for use in feed, malt, and ethanol production.