Profiling LC-DAD-ESI-TOF MS method for the determination of phenolic metabolites from avocado (Persea americana)

A powerful HPLC-DAD-ESI-TOF MS method was established for the efficient identification of the chemical constituents in the methanolic extracts of avocado (Persea americana). Separation and detection conditions were optimized by using a standard mix containing 39 compounds belonging to phenolic acids and different categories of flavonoids, analytes that could be potentially present in the avocado extracts. Optimum LC separation was achieved on a Zorbax Eclipse Plus C18 analytical column (4.6×150 mm, 1.8 μm particle size) by gradient elution with water+acetic acid (0.5%) and acetonitrile as mobile phases, at a flow rate of 1.6 mL/min. The detection was carried out by ultraviolet-visible absorption and ESI-TOF MS. The developed method was applied to the study of 3 different varieties of avocado, and 17 compounds were unequivocally identified with standards. Moreover, around 25 analytes were tentatively identified by taking into account the accuracy and isotopic information provided by TOF MS.     

Hydro-physical responses of gypseous and non-gypseous soils to livestock grazing in a semi-arid region of NE Spain

Pasture productivity depends on soil hydro-physical properties, which in turn are deeply affected by livestock grazing. However, the comparative response of different soil types, and particularly gypseous soil types, to grazing has hardly been studied before. This paper compares the effect of grazing on the soil hydro-physical properties of silty gypseous (Gy) and non-gypseous (NGy) soils located in a semi-arid region (Middle Ebro Valley, NE, Spain). Two different soil managements were selected: ungrazed natural shrubland (N) and grazed shrubland (GR) soils. The gypsum, CaCO₃ and organic matter content (OM), soil texture, soil bulk density (ρ_b), penetration resistance (PR), saturated sorptivity (S), hydraulic conductivity (K), and the water retention curve (WRC) for undisturbed soil samples from 1 to 10 cm depth soil layer were measured. The ρ_b and PR in NGy soils were significantly higher than those observed in the Gy ones. Soil compaction due to grazing treatment tended to increase ρ_b and decrease the K and S values. While no differences in PR were observed in the Gy soils between grazing treatments, the PR measured in the NGy soils under GR was significantly higher than the corresponding values observed under N. Differences in K and S between GR and N treatments were only significant (p < 0.05) in NGy soils, where K and S values under the N treatment were almost four times greater than the corresponding values measured under GR. Overall, no differences in the WRCs were observed between soil types and grazing treatments. While the WRCs of NGy soils were not significantly affected by the grazing treatment, Gy soils under N treatment present a significantly higher level of soil macropores than under GR treatment. The hydro-physical features of Gy soils tended to be less affected by grazing than those of the NGy soils. These results suggest that livestock grazing, in both Gy and NGy soils, has a negative effect on the physical soil properties, which should be taken into account by land managers of these semi-arid regions where silty gypseous and non-gypseous areas coexist.     

Soil variability through spatial scales in a permanently disturbed natural spruce-fir-beech forest

Soil variability was assessed in a 74.2-ha area within the Žofínsky prales natural forest. Parameters evaluated for 1765 soil profiles inside 353 graticule plots were as follows: (1) thickness of organic horizons, (2) thickness and form of mineral horizons, (3) humus form (HF), (4) soil taxonomic unit (STU) and (5) anomalies. In addition, soil reaction (pH_KCl) and oxidizable carbon content (C_ox) were measured in the laboratory for 734 samples from the upper mineral (A) and lower mineral (B) horizons. The most frequently occurring humus form was mor followed by moder, hydromor and peaty T-horizon. Entic Podzols, Dystric Cambisols, Haplic Cambisols, Albic Podzols, Histic (or Haplic) Gleysols, Endogleyic Stagnosols, Fibric or Hemic or Sapric Histosols and Stagnic Gleysols were all present at the site despite its homogeneous geological bedrock. Overall coefficient of variance (CV) was lower in terrestrial soils compared with (semi-)hydromorphic soils. Overall variance decreased in both soil groups with increasing depth, as did CV differences between the fine (up to 10 m) and the locality scales. The lowest CV values occurred for C_ox and pH_KCl. The CV values differed between STUs as well. Compared to lower horizons, variograms of upper horizons showed greater autocorrelation at the intermediate spatial scale (10–320 m)—ranging from 50 to 150 m. Semivariance values, however, reached 70–80% of sill already at a distance of 10 m. The most significant factor of variability at all studied spatial scales is presumably the soil disturbance regime, followed by terrain micro-topography and the effect of tree species.     

Do mature pine plantations resemble deciduous natural forests regarding understory plant diversity and canopy structure in historically modified landscapes?

We compared the structure of the arboreal layer and the diversity and species composition of the understory vegetation of three types of mature forest communities: oak (Quercus pyrenaica) and beech (Fagus sylvatica) forests and Scots pine (Pinus sylvestris) plantations. Our main aim was to determine whether differences in these variables existed and were due to the identity of the dominant tree species. We selected four stands or replicates per forest type located geographically close and with relatively similar conditions. We found no differences in the arboreal structure of oak and beech forests, which were characterised by great variability in tree size, while in case of plantations, this variability was lower at both the intra-stand (estimated by the coefficient of variation) and inter-stand (i.e. the four replicates harboured trees of similar sizes) scales. However, the highest variability in the canopy layer of natural forests was not consistently linked to greater understory species richness. Indeed, the lowest plant species richness was found in beech forests, while oak forests harboured the highest value at either the sampling unit (per m²) or stand scales. The greatest negative correlation between plant diversity and the environmental variables measured was found for litter depth, which was the highest in beech forests. The results obtained by the CCA indicated that the four replicates of each forest type clustered together, due to the presence of characteristic species. We concluded that pine plantations did not approach the environmental conditions of native forests, as plantations were characterised by singular understory species composition and low arboreal layer variability, compared to natural woodlands.     

Susceptibility of the Mexican bean beetle <Emphasis Type="Italic">Epilachna varivestis</Emphasis> Mulsant (Coleoptera: Coccinellidae) to endemic isolates of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae)

Entomopathogenic fungi represent excellent candidates for biological insecticides. Among the many entomopathogenic fungi, Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae) is one of the most promising species. Here, we report the results of infection studies performed using four B. bassiana native isolates (Bb37, Bb38, Bb40, and Bb45) compared with the commercial product Mycotrol^® (strain GHA) on adult Mexican bean beetle Epilachna varivestis Mulsant (Coleoptera: Coccinellidae), one of the most important pests of bean production in Mexico. First, single concentration (1 × 10⁸ conidia mL^?1) assays were carried out using all isolates applied using an immersion method. Examinations at 10 days after inoculation indicated that isolates Bb37, Bb38, and Bb40 significantly reduced adult survival, causing 60–75% mortality compared to the commercial strain GHA (33%) and control (29%). Second, using two selected isolates (Bb37 and Bb40), the mean lethal concentration values (LC₅₀) were estimated for third instar larvae and adult E. varivestis. The LC₅₀ values for Bb37 and Bb40 isolates decreased 5.26-fold and 3.19-fold, respectively, in third instars compared with adults. However, such difference for Bb40 isolate was not significant. Finally, an experiment, to compare the median survival time (MST) values between third instar and adult E. varivestis, was conducted using Bb37 and Bb40 at 1 × 10⁹ conidia mL^?1. MST values were significantly reduced against larvae (87 and 100 h) compared with adults (130 and 134 h) by Bb37 and Bb40, respectively. We conclude that native isolates of B. bassiana represent an important alternative for the control of E. varivestis. However, to demonstrate its effectiveness under field conditions, detailed studies are needed.     

The toxicity of selected acaricides against five stored product mites under laboratory assay

In laboratory tests, the toxicity of acaricides targeted against house dust mites was tested on five species of stored product mites (Acarus siro, Aleuroglyphus ovatus, Carpoglyphus lactis, Lepidoglyphus destructor, and Tyroborus lini). The formulations of benzyl-benzoate, benzyl-benzoate/permethrin/pyriproxyfen, and neem were diluted in water and applied to filter paper in an unventilated chamber. The mortality of mites was observed after 24 h of exposure to acaricide-impregnated filter paper. All of the tested acaricides were toxic to all of the mite species. There were significant differences in mortality among the species and the acaricides. Benzyl-benzoate/permethrin/pyriproxyfen was the most effective, followed by benzyl-benzoate and neem. L. destructor (LD₅₀ 0.01–0.11 μg) was the most sensitive mite species, followed by A. siro (LD₅₀ 0.04–0.12 μg), T. lini (LD₅₀ 2–21 μg), A. ovatus (LD₅₀ 3–18 μg), and C. lactis (LD₅₀ 4–64 μg). Based on the highly toxic effects of the tested acaricides against the stored product mites, the acaricides should be considered as a potential tool in the control of stored product mites, although next screening is necessary.     

Resveratrol oligomers isolated from Carex species inhibit growth of human colon tumorigenic cells mediated by cell cycle arrest

Research has shown that members of the Carex genus produce biologically active stilbenoids including resveratrol oligomers. This is of great interest to the nutraceutical industry given that resveratrol, a constituent of grape and red wine, has attracted immense research attention due to its potential human health benefits. In the current study, five resveratrol oligomers (isolated from Carex folliculata and Carex gynandra ), along with resveratrol, were evaluated for antiproliferative effects against human colon cancer (HCT-116, HT-29, Caco-2) and normal human colon (CCD-18Co) cells. The resveratrol oligomers included one dimer, two trimers, and two tetramers: pallidol (1); α-viniferin (2) and trans-miyabenol C (3); and kobophenols A (4) and B (5), respectively. Although not cytotoxic, the resveratrol oligomers (1-5), as well as resveratrol, inhibited growth of the human colon cancer cells. Among the six stilbenoids, α-viniferin (2) was most active against the colon cancer cells with IC(50) values of 6-32 μM (>2-fold compared to normal colon cells). Moreover, α-viniferin (at 20 μM) did not induce apoptosis but arrested cell cycle (in the S-phase) for the colon cancer but not the normal colon cells. This study adds to the growing body of knowledge supporting the anticancer effects of resveratrol and its oligomers. Furthermore, Carex species should be investigated for their nutraceutical potential given that they produce biologically active stilbenoids such as α-viniferin.     

Effect of aging on lees and of three different dry yeast derivative products on Verdejo white wine composition and sensorial characteristics

A study was made of the effect of aging on lees and of three different commercial yeast derivative products of different composition and degree of purification on the phenolic compounds, color, proteins, polysaccharides, and sensorial characteristics of white wines. The results obtained showed that the lees and yeast derivative products can interact or adsorb some of the phenolic compounds present in wines, reducing their concentration. This reduction depends on the treatment applied, the phenolic compound analyzed, and the stage of vinification or aging process. The use of lees and yeast derivative products can reduce the color intensity and the browning of the wines immediately following treatment. The monosaccharide and polysaccharide content of yeast derivative products depends on the manufacturing process and degree of purification of the product, both of which have an influence on wine treatments. After 6 months in the bottle, both the aging on lees and the treatment with commercial yeast derivative products gave rise to wines with better sensorial characteristics than in the case of the control wines.     

Hepatic immune response in calves during acute subclinical infection with bovine viral diarrhoea virus type 1

Eight colostrum-deprived calves aged 8-12 weeks were inoculated intranasally with a non-cytopathic strain of bovine viral diarrhoea virus (BVDV) genotype-1 and the effects on the hepatic immune response were studied. Two calves were sacrificed at each of 3, 6, 9 and 14 days post-inoculation (dpi) and two uninoculated animals were used as negative controls. BVDV was detected in hepatic macrophages and monocytes from 3 to 14dpi and in Küpffer cells (KCs) from 6 to 14dpi. Increases in the numbers of MAC387(+) KCs and monocytes, but not interstitial macrophages, differentiated by morphological features, were evident in the liver following inoculation with BVDV. There was a substantial increase in the number of monocytes positive for tumour necrosis factor (TNF)-α, but only small increases in the numbers of TNF-α(+) KCs and interstitial macrophages and interleukin (IL)-6(+) monocytes, KCs and interstitial macrophages. There was an increase in the number of interstitial CD3(+) T lymphocytes in the liver, but no substantial changes in the numbers of circulating CD3(+) T lymphocytes, interstitial or circulating CD4(+) or CD8(+) T lymphocytes, or CD79αcy(+) B lymphocytes. Serum haptoglobin and serum amyloid A increased transiently at 12dpi. Upregulation of some pro-inflammatory cytokines by hepatic macrophages is evident in subclinical acute BVDV type 1 infection in calves.